CA2583428A1 - Bicyclic heteroaryl pde4b inhibitors - Google Patents

Abstract

Compounds are described that are active on PDE4. Also described are crystal structures of PDE4B determined using X-ray crystallography, the use of PDE4B
crystals and stractural information for identifying molecular scaffolds, for developing ligands that bind to and modulate PDE4B, and for identifying improved ligands based on known ligands.

Description

DEMANDE OU BREVET VOLUMINEUX

LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET COMPREND
PLUS D'UN TOME.

NOTE : Pour les tomes additionels, veuillez contacter le Bureau canadien des brevets JUMBO APPLICATIONS/PATENTS

THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE THAN ONE
VOLUME

NOTE: For additional volumes, please contact the Canadian Patent Office NOM DU FICHIER / FILE NAME:

NOTE POUR LE TOME / VOLUME NOTE:

FIELD OF THE INVENTION

[0001] This invention relates to the development of ligands for phosphodiesterase PDE4, including for PDE4 isoforms PDE4B and PDE4D, and to the use of crystal structures of PDE4B and/or PDE4D. Additionally, this invention provides coinpounds with activity toward PDE4, and methods of use thereof.

BACKGROUND OF THE INVENTION

[0002] The information provided is intended solely to assist the understanding of the reader.
None of the information provided nor references cited is admitted to be prior art to the present invention. Each of the references cited is incorporated herein in its entirety.

[0003] Phosphodiesterases (PDEs) were first detected by Sutherland and co-workers (Rall, et al., J. Biol. Chem., 232:1065-1076 (1958), Butcher, et al., J. Biol. Chem., 237:1244-1250 (1962)). The superfamily of PDEs is subdivided into two major classes, class I
and class II
(Charbonneau, H., Cyclic Nucleotide Phosphodiesterases: Structure, Regulation and Drug Action, Beavo, J., and Houslay, M.D., eds) 267-296 John Wiley & Sons, Inc., New York (1990)), which have no recognizable sequence similarity. Class I includes all known mammalian PDEs and is coinprised of 11 identified families that are products of separate genes (Beavo, et al., Mol. Pharmacol., 46:399-405 (1994); Conti, et al., Endocr.
Rev., 16:370-389 (1995); Degeiman, et al., J. Biol. Clzem., 272:6823-6826 (1997); Houslay, M.D., Adv. Enzyme Regul., 35:303-338 (1995); Bolger, G.B., Cell Signal, 6:851-859 (1994);
Thompson, et al, Adv.
Second Messenger Phosphoprotein Res., 25:165-184 (1992); Underwood, et al., J.
Pharnzacol.
Exp. Ther., 270:250-259 (1994); Michaeli, et al., J. Biol. Chem., 268:12925-12932 (1993);
Soderling, et al., Proc. Natl. Acad. Sci. U.S.A., 95:8991-8996 (1998);
Soderling, et al., J. Biol.
Chem., 273:15553-15558 (1998); Fisher, et al., J Biol. Claem., 273:15559-15564 (1998)).
Some PDEs are highly specific for hydrolysis of cAMP (PDE4, PDE7, PDE8), some are highly cGMP-specific (PDE5, PDE6, PDE9), and some have mixed specificity (PDE1, PDE2, PDE3, PDE10, PDE1 1) (Conti, Molecular Endocrinology, 14:1317-1327 (2000)).

[0004] All of the characterized mainmalian PDEs are dimeric, but the importance of the dimeric structure for function in each of the PDEs is unknown. Each PDE has a conserved catalytic domain of -270 amino acids with a high degree of conservation (25-30%) of amino acid sequence among PDE families, which is located carboxyl-terminal to its regulatory domain. Activators of certain PDEs appear to relieve the influence of autoinhibitory domains located within the enzyme structures (Sonnenberg, et al., J. Biol. Chem., 270:30989-31000 (1995); Jim, et al., J. Biol. Chein., 267:18929-18939 (1992)).

[0005] PDEs cleave the cyclic nucleotide phosphodiester bond between the phosphorus and oxygen atoms at the 3'-position with inversion of configuration at the phosphorus atom (Goldberg, et al., J. Biol. Chem., 255:10344-10347 (1980); Burgers, et al., J.
Biol. Claem., 254:9959-9961 (1979)). This apparently results from an in-line nucleophilic attack by the OH-of ionized H20. It has been proposed that metals bound in the conserved metal binding motifs within PDEs facilitate the production of the attacking OH- (Francis, et al., J. Biol. Chem., 269:22477-22480 (1994)). The kinetic properties of catalysis are consistent witll a random order mechanism with respect to cyclic nucleotide and the divalent cations(s) that are required for catalysis (Srivastava, et al., Biochem. J, 308:653-658 (1995)). The catalytic domains of all known mammalian PDEs contain two sequences (HX3 HXõ(E/D)) arranged in tandem, each of which resembles the single Zn2+-binding site of metalloendoproteases such as thermolysin (Francis, et al., J. Biol. Chein., 269:22477-22480 (1994)). PDE5 specifically binds Zn2+, and the catalytic activities of PDE4, PDE5, and PDE6 are supported by submicromolar concentrations of ZnZ+ (Francis, et al., J. Biol. Chem., 269:22477-22480 (1994); Percival, et al., Biochem. Biophys. Res. Cominun., 241:175-180 (1997)). Whether each of the ZnZ+-binding motifs binds Zn2+ independently or whether the two motifs interact to form a novel Zn2+-binding site is not known. The catalytic mechanism for cleaving phosphodiester bonds of cyclic nucleotides by PDEs may be siniilar to that of certain proteases for cleaving the amide ester of peptides, but the presence of two ZnZ+ motifs arranged in tandem in PDEs is unprecedented.

[0006] The group of Sutherland and Rall (Berthet, et al., J Biol. Clzena., 229:351-361 (1957)), in the late 1950s, was the first to realize that at least part of the mechanism(s) whereby caffeine enhanced the effect of glucagon, a stimulator of adenylyl cyclase, on cAMP
accumulation and glycogenolysis in liver involved inhibition of cAMP PDE
activity. Since that time chemists have synthesized thousands of PDE inhibitors, including the widely used 3-isobutyl-l-methylxanthine (IBMX). Many of these compounds, as well as caffeine, are non-selective and inhibit many of the PDE families. One important advance in PDE
research has been the discovery/design of family-specific inhibitors such as the PDE4 inhibitor, rolipram, and the PDE5 inhibitor, sildenafil.

[0007] Precise modulation of PDE function in cells is critical for maintaining cyclic nucleotide levels within a narrow rate-limiting range of concentrations.
Increases in cGMP of 2-4-fold above the basal level will usually produce a maximum physiological response. There are three general schemes by which PDEs are regulated: (a) regulation by substrate availability, such as by stimulation of PDE activity by mass action after elevation of cyclic nucleotide levels or by alteration in the rate of lzydrolysis of one cyclic nucleotide because of competition by another, which can occur with any of the dual specificity PDEs (e.g. PDE1, PDE2, PDE3); (b) regulation by extracellular signals that alter intracellular signaling (e.g.
phosphorylation events, Ca2+, phosphatidic acid, inositol phosphates, protein-protein interactions, etc.) resulting, for exainple, in stimulation of PDE3 activity by insulin (Degerman, et al., J. Biol. Chem., 272:6823-6826 (1997)), stimulation of PDE6 activity by photons through the transducin system (Yamazaki, et al., J. Biol. Chem., 255:11619-11624 (1980)), which alters PDE6 interaction with this enzyme, or stimulation of PDE1 activity by increased interaction with Ca2+/calmodulin; (c) feedback regulation, such as by phosphorylation of PDE1, PDE3, or PDE4 catalyzed by PKA after cAmP elevation (Conti, et al., Endocr. Rev., 16:370-389 (1995);
Degerman, et al., J. Biol. Chem., 272:6823-6826 (1997); Gettys, et al., J.
Biol. Chem. 262:333-339 (1987); Florio, et al, Biochenaistry, 33:8948-8954 (1994)), by allosteric cGMP binding to PDE2 to promote breakdown of cAMP or cGMP after cGMP elevation, or by modulation of PDE protein levels, such as the desensitization that occurs by increased concentrations of PDE3 or PDE4 following clironic exposure of cells to cAMP-elevating agents (Conti, et al., Endocr. Rev., 16:370-389 (1995), Sheth, et al., Throm. Haemostasis, 77:155-162 (1997)) or by developmentally related changes in PDE5 content. Other factors that could influence any of the three schemes outlined above are cellular compartmentalization of PDEs (Houslay, M.D., Adv. Enzyme Regul., 35:303-338 (1995)) effected by covalent modifications such as prenylation or by specific targeting sequences in the PDE primary structure and perhaps translocation of PDEs between compartments within a cell.

[0008] Within the PDE superfamily, four (PDE2, PDE5, PDE6, and PDE10) of the families contain highly cGMP-specific allosteric (non-catalytic) cGMP-binding sites in addition to a catalytic site of varying substrate specificity. Each of the monomers of these dimeric cGMP-binding PDEs contains two homologous cGMP-binding sites of - 110 amino acids arranged in tandem and located in the amino-terminal portion of the protein (Charbonneau, H., Cyclic Nucleotide Plzosphodiesterases: Structure, Regulation and Drug Action, Beavo, J., and Houslay, M.D., eds) 267-296 (1990); McAllister-Lucas, et al., J. Biol.
Chem., 270:30671-30679 (1995)). In PDE2, binding of the cGMP to these sites stimulates the hydrolysis of cAMP at the catalytic site (Beavo, et al., Mol. Pharmacol., 46:399-405 (1994)).
PDE2 hydrolyzes cGMP as well as cAMP, and cGMP hydrolysis is stimulated by cGMP
binding at the allosteric sites in accordance with positively cooperative kinetics (Manganiello, et al., Cyclic Nucleotide Phosphodiesterases: Structure, Regulation, and Drug Action, Beavo, J., and Houslay, M.D., eds, 61-85 John Wiley & Sons, Inc., New York (1990)).
This could represent a negative feedback process for regulation of tissue cGMP levels (Manganiello, et al., Cyclic Nucleotide Phosphodiesterases: Structure, Regulation, and Drug Action, Beavo, J., and Houslay, M.D., eds, 61-85 John Wiley & Sons, Inc., New York (1990)), which occurs in addition to the cross-talk between cyclic nucleotide pathways represented by cGMP
stimulation of cAMP breakdown. Binding of cGMP to the allosteric sites of PDE6 has not been shown to affect catalysis, but this binding may modulate the interaction of PDE6 with the regulatory protein, transducin, and the inhibitory y subunit of PDE6 (Yamazaki, et al., Adv.
Cyclic Nucleotide Protein Phosphofylation Res., 16:381-392 (1984)).

[0009] The PDE4 subfamily is comprised of 4 members: PDE4A, PDE4B (SEQ ID
NO:1), PDE4C, and PDE4D (SEQ ID NO:2) (Conti et al. (2003) JBiol Ch.ena. 278:5493-5496). The PDE4 enzymes display a preference for cAMP over cGMP as a substrate. These enzymes possess N-terminal regulatory domains that presumably mediate dimerization, which results in optimally regulated PDE activity. In addition, activity is regulated via cAMP-dependent protein kinase phosphorylation sites in this upstream regulatory domain. These enzymes are also rather ubiquitously expressed, but importantly in lymphocytes.

[0010] Inliibitors of the PDE4 enzymes have proposed utility in the treatment of inflammatory diseases. Knockout of PDE4B results in viable mice (Jin and Conti (2002) Proc Natl Acad Sci U S A, 99, 7628-7633), while knockout of PDE4D results in reduced viability (Jin et al. (1999) Proc Natl Acad Sci USA, 96, 11998-12003). The PDE4D
knockout genotype can be rescued by breeding onto other background mouse strains.
Airway epithelial cells from these PDE4D knockout embryos display greatly reduced hypersensitivity to adrenergic agonists, suggesting PDE4D as a therapeutic target in airway inflammatory diseases (Hansen et al. (2000) Proc Natl Acad Sci U S A, 97, 6751-6756). PDE4B-knockout mice have few symptoms and normal airway hypersensitivity. Delgado et al., (MedSciMonit, 2003, 9:BR252-259), report that in view of the increase in PDE4 activity described in blood mononuclear white cells of patients with atopic dermatitis, and the putative relationship between histamine and PDE4 in inflammatory cells, histamine up-regulates PDE4 activity in U-937 cells through H2 receptor stimulation and cAMP increase.

[0011] By contrast, monocytes from the PDE4B knockout mice exhibit a reduced response to LPS (Jin and Conti (2002) Proc Natl Acad Sci U S A, 99, 7628-7633). This suggests that a PDE4B compound with selectivity versus PDE4D could exhibit anti-inflammatory activity with reduced side-effects.

[0012] Accordingly, there is a need in the art for more potent and specific inhibitors and modulators of PDE4 such as PDE4B and PDE4D and methods for designing them.
SUMMARY OF THE INVENTION

[0013] The present invention provides compounds active on PDE4, e.g., PDE4B
and/or PDE4D isofonns. In particular, the invention provides compounds of Formula I, Ia, Ib, Ic, Id, and Ie as described below. Thus, the invention provides compounds that can be used for therapeutic methods involving modulation of PDE4, as well as providing molecular scaffolds for developing additional modulators of PDE4, and other PDEs. The invention further involves the use of structural information about PDE4B to derive additional modulators.

[0014] In one aspect, the invention provides compounds of Formula I having the following structure:

v~ \
x u y N

Formula I

wherein:
k is selected from the group consisting of -CR6R7R19, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12 R13, and -S(O)2R14;

Z is O, S, or NR9;
t, u, v, w, x, and y are each independently N or CRI, provided, however, that no more than one of u and t are N, and no more than two of v, w, x and y are N;
A is selected from the group consisting of substituted aryl and substituted heteroaryl;
Rl at each occurrence is independently selected from the group consisting of hydrogen, halogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylakyl, optionally substituted aralkyl, optionally substituted heteroaralkyl, -C(Z)R8, -OR9, -SR9, -NR1oR11, -C(Z)NR12 R13, -S(O)2NR12 R13, -S(O)2R14, and A, provided, however, that at least one R' is A;
R6 and R7 are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylakyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R8 at each occurrence is independently selected from the group consisting of -OR9, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R$ is alkenyl, no alkene carbon thereof is bound to C(Z), optionally substituted lower alkynyl, provided, however, that when R8 is alkynyl, no alkyne carbon thereof is bound to C(Z), optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R9 at each occurrence is independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R9 is alkenyl, no alkene carbon thereof is bound to 0, N or S, optionally substituted lower alkynyl, provided, however, that when R9 is alkynyl, no alkyne carbon thereof is bound to 0, N or S, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R10 and Rll at each occurrence are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R10 and/or R11 are alkenyl, no alkene carbon thereof is bound to nitrogen, optionally substituted lower alkynyl, provided, however, that when R10 and/or Rl1 are alkynyl, no alkyne carbon thereof is bound to nitrogen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, -C(Z)NR12R13, -S(0)2NR12 R13, and -S(0)2R14; or R10 and Ril together with the nitrogen to which they are attached form a 5-7 meinbered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring;
R12 and R13 at each occurrence are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R12 and/or R13 are alkenyl, no alkene carbon thereof is bound to nitrogen, optionally substituted lower alkynyl, provided, however that when R12 and/or R13 are alkynyl, no alkyne carbon thereof is bound to nitrogen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl; or R12 and R13 together with the nitrogen to which they are attached form a 5-7 membered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring;
R14 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R14 is alkenyl, no alkene carbon thereof is bound to -S(0)2-, optionally substituted lower alkynyl, provided, however, that when R14 is alkynyl, no alkyne carbon thereof is bound to -S(O)z-, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R19 is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, or optionally substituted heteroaralkyl; and all salts, prodrugs, tautomers and isomers thereof.

[0015] In one embodiment of the invention, no more than one of t, u, v, w, x, and y of Formula I is N. In one embodiment, t is N or CH, provided that no more than one of t, u, v, w, x, and y is N. In one einbodiment, t, u, v, w, and x are CR1, and y is N or CRI; and in a further embodiment, t is CH. In one embodiment, t is N or CH, y is N or CH, provided t and y are not both N, one of u, v, w, and x is C-A, and the others of u, v, w and x are CH, provided that the compound is not ~ \ ~ \ p \
~

~N (;N /N N
N N
O-\

F

_ _ N

\ / ,or \ /.

In one embodiment of the invention, t is CH, y is N, one of u, v, w, and x is C-A, and the others of u, v, w and x are CH, provided that the compound is not F

bi ~
N N -02S ~ ~

[0016] Further to any of the above embodiments, A of Formula I is selected from the group consisting of substituted phenyl, dialkoxyphenyl, pyrazole carboxylic ester, substituted pyridine, substituted pyrimidine, and substituted thienopyriinidine. In one embodiment, A has a structure of one of the following groups, in which the squiggle line indicates the attachment to the bicyclic core structure of Formula I.

R15 R16/p R16~

O O
N

o O IS

R16/ R17S R17 li \
II I ~
O
~
, > >
O

R16/O R16 \
N/
O N / ~ ~N
_N -O
, > >
R'$

N' N
ji "~' N
R17/ i S, ~ R1s N~ ts' s > > >

R17___. o R17_o O
O
, or wherein:
R15 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R15 is alkenyl, no alkene carbon thereof is bound to oxygen, optionally substituted lower alkynyl, provided, however, that when R15 is alkynyl, no alkyne carbon thereof is bound to oxygen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, and -C(Z)NR12R13; or both R15s along witli the oxygens to which they are bound combine to form a 5-membered optionally substituted heterocycloalkyl ring fused to the phenyl ring;
R16 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R16 is alkenyl, no alkene carbon thereof is bound to oxygen, optionally substituted lower alkynyl, provided, however, that when R16 is alkynyl, no alkyne carbon thereof is bound to oxygen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, and -C(Z)NR12R13;

R17 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl provided, however, that when Rl7 is alkenyl, no alkene carbon thereof is bound to N, 0, or S, optionally substituted lower allcynyl, provided, however, that when R17 is alkynyl, no alkyne carbon thereof is bound to N, 0, or S, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;

Rl$ at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl, -C(Z)R8, -OR9, -SR9, -NRl Rll, -C(Z)NR12R13, -S(O)2NR1ZR13, or -S(O)ZR14; and all salts, prodrugs, tautomers and isomers thereof.

[0017] Further to any of the above embodiments, k of compounds of Formula I is selected from the group consisting of -CH2R19, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12R13, and S O 14 wherein R8, RlZ > R 13 > R 14 >
- and R19 are selected from the group consisting of ( )2R >
optionally substituted lower alkyl, aryl and heteroaryl, wherein aryl and heteroaryl are optionally substituted with 1-3 substituents selected from the group consisting of halogen, optionally substituted lower alkyl, optionally substituted lower alkoxy, optionally substituted lower thioalkyl, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted amino, optionally substituted carboxyl, optionally substituted alkylsulfonylamino, cyano'und nitro.

[0018] In certain embodiments of the invention, the compoiulds of Formula I
have a structure of Formula Ia:

v~W\
A X

y N

Formula Ia wherein t, v, w, x, y, k and A are as defined in Formula I above; and all salts, prodrugs, tautomers and isomers thereof.

[0019] In one embodiment, A of compounds of Formula Ia has a structure of one of the following groups, in wliich the squiggle line indicates the attachment to the bicyclic core structure of Formula Ia.

/ I
o ~

~ ~

, 0 o Is li O

O

I I / N/
O N N
~N ~--0 I,n R'$

N
~ I N
R17 g sss N
R 17 R~$

O , or wherein Rls, R16, R17 and R18 are as defined in Formula I above.

[0020] In another embodiment of the invention, no more than one of t, v, w, x, and y is N. In one embodiment, t is N or CH, provided that no more than one of t, u, v, w, x, and y is N. In one embodiment, t, u, v, w, and x are CR', and y is N or CRI; and in a further embodiment, t is CH. In another embodiment, t is N or CH, y is N or CH, provided t and y are not both N, two of v, w, and x are CR', and the other of v, w and x is CH. In another embodiment, t is N or CH, y is N or CH, provided t and y are not both N, one of v, w, and x is CRl and the others of v, w and x are CH. In another embodiment, t is N or CH, y is N or CH, provided t and y are not both N, and v, w, and x are CH. In another embodiment, t, v, w, x and y are CH. In another embodiment, t, v, w and x are CH, and y is N. In another embodiment, t and y are CH, one of v, w, and x is CR' and the others of v, w and x are CH. In another embodiment, t is CH, y is N, one of v, w, and x is CRl and the others of v, w and x are CH. Further to any of the above embodiments of Formula Ia, the compound is not O~
~O

~
\
(~ N
i N ' O

[0021] In one eindodiment, k of compounds of Formula Ia is selected from the group consisting of -CH2R19, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14, wherein R8, Rla, R13, R14, and R19 are selected from the group consisting of optionally substituted lower alkyl, aryl and heteroaryl, wherein aryl and heteroaryl are optionally substituted with 1-3 substituents selected from the group consisting of halogen, optionally substituted lower alkyl, optionally substituted lower alkoxy, optionally substituted lower thioalkyl, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted amino, optionally substituted carboxyl, optionally substituted alkylsulfonylamino, cyano and nitro.

[0022] In certain embodiments of the invention, the compounds of Formula I
have a structure of Formula Ib:
A

x u \ y~
--- \
t, N

k Formula lb wherein t, u, w, x, y, k and A are as defined in Formula I above; and all salts, prodrugs, tautomers and isomers thereof.

[0023] In one embodiment, A of compounds of Formula Ib has a structure of one of the following groups, in which the squiggle line indicates the attachment to the bicyclic core structure of Formula Ib.

R16/O R16~
/O \ ~ 0 O

o 0 IS

R16 \ R17/ S R17 II I
II
o O

\ O/
R16/O g RO N
/ O ~ N y~ ~ N

~N L-O I n , > >

N ' ~ ji N

5ss' R~~ R~s N
, > >

O
O , or wherein Rls, R16, R17 and Rl$ are as defined in Formula I above.

[0024] In another embodiment of the invention, no more than one of t, u, w, x, and y is N.
In one embodiment, t is N or CH, provided that no more than one of t, u, v, w, x, and y is N. In one embodiment, t, u, v, w, and x are CRI, and y is N or CRI; and in a further embodiment, t is CH. In another embodiment, t is N or CH, y is N or CH, provided t and y are not both N, two of u, w, and x are CRI, and the other of u, w and x is CH. In another embodiment, t is N or CH, y is N or CH, provided t and y are not both N, one of u, w, and x is CRl and the others of u, w and x are CH. In another embodiment, t is N or CH, y is N or CH, provided that t and y are not both N, and u, w, and x are CH. In another einbodiment, t, u, w, x and y are CH. In another embodiment, t is CH, y is N, one of u, w, and x is CR' and the otllers of u, w and x are CH. In another embodiment, t, u, w and x are CH and y is N. In another embodiment, t a.nd y are CH, one of u, w, and x is CR' and the others of u, w and x are CH. Further to any of the above embodiments of Formula Ib, the compound is not o-\

I \
N
OS \ /

[0025] In one emdodiment of the invention, k of compounds of Formula lb is selected from the group consisting of -CHZR19, -C(Z)R8, -C(Z)NR12R13, -S(O)ZNR12R13, and -S(O)2R14, wherein R8, R12, R13, R14, and R19 are selected from the group consisting of optionally substituted lower alkyl, aryl and heteroaryl, wherein aryl and heteroaryl are optionally substituted with 1-3 substituents selected from the group consisting of halogen, optionally substituted lower alkyl, optionally substituted lower alkoxy, optionally substituted lower thioalkyl, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted amino, optionally substituted carboxyl, optionally substituted alkylsulfonylamino, cyano and nitro.

[0026] In certain embodiments, the compounds of Fonnula I have a structure of Formula Ic:
A

v4--x y t~ N
k Formula Ic wherein t, u, v, x, y, k and A are as defined in Formula I above; and all salts, prodrugs, tautomers and isomers thereof.

[0027] In one embodiment, A of compounds of Formula Ic has a structure of one of the following groups, in which the squiggle line indicates the attachment to the bicyclic core structure of Formula Ic.
O
R~s/0 R16~

N

OI
O lO I
R16/ S! R17 O / O

> > >
O

Ri s/O R16 )91 N/ O N y ~N

~N '--O

R'$

N
N
N / I
R17/ S S~
~s' R~$ N ~

R17_ o O
O , or wherein Rls, R16, R17 and R18 are as defined in Formula I above.

[0028] In another embodiment of the invention, no more than one of t, u, v, x, and y is N. In one embodiment, t is N or CH, provided that no more than one of t, u, v, w, x, and y is N. In one embodiment, t, u, v, w, and x are CRI, and y is N or CRI; and in a further embodiment, t is CH. In another embodiment, t is N or CH, y is N or CH, provided t and y are not both N, two of u, v, and x are CRI, and the other of u, v and x is CH. In another embodiment, t is N or CH, y is N or CH, provided t and y are not both N, one of u, v, and x is CR' and the others of u, v and x are CH. In another embodiment, t is N or CH, y is N or CH, provided t and y are not both N, and u, v, and x are CH. In another embodiment, t, u, v, x and y are CH. In another embodiment, t, u, v and x are CH and y is N. In another embodiment, t and y are CH, one of u, v, and x is CR1 and the others of u, v and x are CH. In another embodiment, t is CH, one of u, v, and x is CR1, the others of u, v, and x are CH, and y is N or CH, provided, however, that when the coinpound has the structure \ \ ~
N N -02S ~ ~

then Rl is selected from the group consisting of hydrogen, F, Cl, Br, optionally substituted lower alkyl, -C(Z)R8, -OR9, -SR9, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)ZR14.

[0029] In one emdodiment of the invention, k of compounds of Formula Ib is selected from the group consisting of -CH2R'9, -C(Z)Rg, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14, wherein R8, R12, R13, R14, and R19 are selected from the group consisting of optionally substituted lower alkyl, aryl and heteroaryl, wherein aryl and heteroaryl are optionally substituted with 1-3 substituents selected from the group consisting of halogen, optionally substituted lower alkyl, optionally substituted lower alkoxy, optionally substituted lower thioalkyl, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted amino, optionally substituted carboxyl, optionally substituted alkylsulfonylamino, cyano and nitro.

[00301 In certain embodiments of the invention, the compounds of Formula I
have a structure of Formula Id:
A
\
N
N
k Formula Id wherein k is as defined in Formula I above and A has a structure of one of the following groups, in which the squiggle line indicates the attachment to the bicyclic core structure of Formula Id:

~ O

~

O
~ Y
> > >
O
~~ ~s O

(I
O / I

> > >

O

\--N

N
N
1 \ j~

R1$
~ , R17 e R17-O

O
O , or wherein Ris, R16, R 17 and R'8 are as defined in Formula I above; and all salts, prodrugs, tautomers and isomers thereof.

[0031] In one embodiment of the invention, k of compounds of Forinula Id is selected from the group consisting of -CH2R19, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14, wherein R8, R12, R13, R14, and R19 are selected from the group consisting of optionally substituted lower alkyl, aryl and heteroaryl, wherein aryl and heteroaryl are optionally substituted with 1-3 substituents selected from the group consisting of halogen, optionally substituted lower allcyl, optionally substituted lower alkoxy, optionally substituted lower thioalkyl, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted amino, optionally substituted carboxyl, optionally substituted alkylsulfonylamino, cyano and nitro.

[0032] In certain embodiments of the invention, the compounds of Formula I
have a structure of Formula le:

O~

O

I \ \
N
N
k Formula le wherein k and R15 are as defined in Formula I above; and all salts, prodrugs, tautomers and isomers thereof.

[0033] In one embodiment of the invention, k of compounds of Formula Ie is selected from the group consisting of -CH2R19, -C(Z)R8, -C(Z)NR1ZR13, -S(O)2NR12R13, and -S(O)2R14, wherein R8, R12, R13, Ri4, and R19 are selected from the group consisting of optionally substituted lower alkyl, aryl and heteroaryl, wherein aryl and heteroaryl are optionally substituted with 1-3 substituents selected from the group consisting of halogen, optionally substituted lower alkyl, optionally substituted lower alkoxy, optionally substituted lower tliioalkyl, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted amino, optionally substituted carboxyl, optionally substituted alkylsulfonylamino, cyano and nitro, further wherein R15 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl, or each R15 along with the oxygens to which they are bound combine to form a 5-7 membered optionally substituted heterocycloalkyl ring fused to the phenyl ring. In a further embodiment, when R15 is optionally substituted lower alkyl, the alkyl is optionally substituted with 1-3 substituents selected from the group consisting of fluoro, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl.

[0034] In certain embodiments of the invention involving compounds of Formula I (for example, compounds of Formula Ia, Formula Ib, and Formula Ic), the bicyclic ring structure shown in Formula I is one of the following:

II
N

N N (-11N II I

N/ N N N
N N
N
N
N \ \ / \ / \ N
N N N
N ~ N N N~ N N

N N N N \ N
\ \ I \

N N N~ N N N
> > > ~
N~N rN
\ \ I
I N
\ N N N N

N::::I I ~ NN ~ , N , QN
N N N
N~ N r N I N N N
/ N N
N N N N N

N/ N N
I ~ N I ~
N N N N
N/N N rN N

N N N N N

N~ N O'N I \
N I N \
\ N
~ N or N

[0035] For compounds of Formula I, ring positions are specified as shown in the following indole core, but as used herein, the corresponding numbering also applies to each of the other bicyclic core structures shown above. Reference to a bicyclic core as in Formula I or indication that a compound includes a bicyclic core as in Formula I and phrases of similar import refer to a bicyclic structure or moiety as described herein for einbodiments of the invention embracing compounds of Formula I.

6 ~ $ 1 N
[0036] In particular embodiments of the invention, compounds of Formula I
(e.g., Formula Ia, Formula Ib, Formula Ic, Formula Id, and Formula Ie) have a substituent at the 1-position as a compound in the Examples; has a substituent at the 3-position as a compound in the Examples; has a substitutent at the 4-position as a compound in the Examples;
has a substitutent at the 5-position as a compound in the Examples; has substitutents at the 1- and 3-positions as a compound in the Examples; has substitutents at the 1- and 4-positions as a compound in the Examples; has substitutents at the 1- and 5-positions as a compound in the Examples.

[0037] In particular embodiments of the invention, one or more of the bicyclic structures shown above are embraced by Formula I, e.g., any 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33 of the bicyclic core structures shown above are embraced by Formula I.

[0038] In certain embodiments of the invention, A of compounds of Formula I
(e.g., Formula Ia, Formula Ib, Formula Ic, and Formula Id) is substituted aryl (e.g. phenyl);
substituted heteroaryl (such as 5- or 6-membered heteroaryl rings); aryl (e.g., phenyl) substituted with 1, 2, or 3 alkoxy groups (such as methoxy, ethoxy, and propoxy); heteroaryl substituted with 1, 2, or 3 alkoxy groups; aryl substituted with 1, 2, or 3 alkyl ether groups, alkyl thioether groups, or coinbinations thereof; heteroaryl substituted with 1, 2, or 3 alkyl ether groups, alkyl thioether groups, or combinations thereof; dialkoxyphenyl; dialkylthiophenyl; dialkoxy heteroaryl;
dialkylthio heteroaryl; disubstituted aryl (e.g., phenyl) in which the substitutents may be the same or different, e.g., hydroxyl, alkoxy, alkyl ester, -SH, thioether, thioester; disubstituted heteroaryl in which the substitutents maybe the same or different, e.g., hydroxyl, alkoxy, alkyl ester, -SH, thioetlier, thioester. In certain embodiments in which an aryl or heteroaryl is disubstituted, the substitutents are on adjacent ring carbon atoms (e.g., catechols or catechol diethers), for 6-meinbered rings the two substituents may be meta and para with respect to the linkage of the ring to the remainder of the molecule. In certain embodiments, A is substituted aryl or substituted heteroaryl, with a substitution at the second atom away from the ring atom attached to the remainder of the molecule (e.g., the meta position for 6-membered rings), in certain embodiments, that is the only substitution on the aryl or heteroaryl group.

[0039] In certain embodiments of the invention, k of compounds of Formula I
(e.g., Formula Ia, Formula Ib, Formula Ic, Formula Id and Formula le) comprises optionally substituted aryl;
optionally substituted heteroaryl; optionally substituted napthyl; optionally substituted bicyclic heteroaryl; napthyl substituted with alkoxy or alkylthio; heteroaryl substituted with alkoxy or alkylthio. For example R6, R7, R8, R9, R12, R13 or R14 in k is a group so specified. In particular embodiments of the preceding, the alkyl moiety is methyl, ethyl, or propyl.

[0040] In particular embodiments, the combination of A and k embraces any A
(as specified in [0038]) and any k (as specified in [0039]), e.g., A is substituted phenyl and k comprises optionally substituted napthyl; A is substituted 5- or 6-membered heteroaryl and k comprises optionally substituted napthyl; A is substituted phenyl and k comprises optionally substituted bicyclic heteroaryl; A is substituted 5- or 6-membered heteroaryl and k comprises optionally substituted bicyclic heteraryl, and the like.

[0041] In certain embodiments of the invention, compounds are excluded where N, 0, S or C(Z) would be bound to a carbon that is also bound to N, O, S, or C(Z) or would be bound to an alkene carbon atom of an alkenyl group or bound to an alkyne atom of an alkynyl group;
accordingly, in certain embodiments compounds are excluded from the present invention in which are included linkages such as -NR-CH2-NR-, -0-CH2-NR-, -S(O)o_2-CH2-NR-, -C(Z)-CH2-NR-, NR-CH2-O-, -O-CHZ-O-, -S(0)0_2-CH2-0-, -C(Z)-CH2-O-, -NR-CHa-S(O)o_2-, -0-CH2-S(O)0_2-, -S(O)0_2-CH2-S(O)0_2-, -C(Z)-CH2-S(O)0_2-, -NR-CH2-C(Z)-, -O-CH2-C(Z)-, -S(O)0_2-CH2-C(Z)-, -C(Z)-CHZ-C(Z)-, NR-CH=CH-, -NR-C=C-, -O-CH=CH-, -O-C=C-, -S(O)o_Z-CH=CH-, -S(O)o_Z-C=C-, -C(Z)-CH=CH-, or -C(Z)-C C-.

[0042] Thus, in a first aspect, the invention provides a novel compound of Formulae I, Ia, Ib, Ic, Id, or le as described herein.

[0043] An additional aspect of this invention provides compositions, which include pharmaceutical formulations, that include a therapeutically effective amount of a compound of Formula I, Ia, Ib, Ic, Id, or Ie (or a compound within a sub-group of compounds within any of those generic formulae) and at least one pharmaceutically acceptable carrier, excipient or diluent.

[0044] In particular embodiments, the composition includes a plurality of different pharmacalogically active compounds, which can be a plurality of compounds of Formula I and can also include other compounds in combination with one or more compounds of Formula I.
The term "other compounds" in this context denotes compounds that are given to a subject in an effective amount to prevent, alleviate, or ameliorate one or more symptoms of a disease or medical condition, and/or to prolong the survival of the subject being treated, wherein the disease or medical condition is as listed herein.

[0045] In a related aspect, the invention provides kits that include a composition as described herein. In particular embodiments, the composition is packaged, e.g., in a vial, bottle, or flask, which may be further packaged, e.g., within a box, envelope, or bag; the composition is a pharmaceutical composition approved by the U.S. Food and Drug Administration or similar regulatory agency for administration to a mammal, e.g.,a human; the composition is a pharmaceutical composition approved for administration to a mammal, e.g., a human, for a PDE4-mediated disease or condition; the kit includes written instructions or other indication that the composition is suitable or approved for administration to a mamtnal, e.g., a human, for a PDE4-mediated disease or condition; the composition is packaged in unit does or single dose form, e.g., single dose pills, capsules, or the like. As used herein, the term "mammal"
indicates any mammalian species, and include without limitation, stock animals (e.g., sheep, goats, cattle), domesticated animals (e.g., dogs, cats), research animals (e.g., rats, mice), other primates, and humans.

[0046] In another related aspect, compounds of Formula I, Ia, Ib, Ic, Id, or le can be used in the preparation of a medicament for the treatment in a subject in need thereof of a PDE4-mediated disease or condition (or a disease or condition mediated by a PDE4 isoform (e.g., a PDE4B- or PDE4D-mediated disease or condition), or a disease or condition in which modulation of PDE4 provides a therapeutic benefit.

[0047] In another aspect, the invention provides a method of treating or prophylaxis of a disease or condition in a mammal in need thereof where the disease or condition is a PDE4-mediated disease or condition or a disease or condition in which PDE4 modulation provides a therapeutic benefit, by administering to the mammal a therapeutically effective amount of a compound of Formula I, Ia, Ib, Ic, Id, or le, a prodrug of such compound, or a pharmaceutically acceptable salt of such compound or prodrug. The compound can be employed alone or can be part of a pharmaceutical composition.

[0048] In aspects and embodiments of the invention involving treatinent or prophylaxis of a disease or condition, the disease or condition is, for example without limitation, an acute or chronic pulmonary disease such as obstructive diseases (e.g. astluna, chronic obstructive pulmonary disease (COPD), cystic fibrosis), interstitial lung diseases (e.g.
idiopathic pulmonary fibrosis, sarcoidosis), vascular lung diseases (e.g. pulmonary hypertension), bronchitis, allergic bronchitis, or emphysema. Additional diseases or conditions contemplated for treatment by embodiments of the present invention include, for example without limitation, CNS diseases such as Alzheimer's disease, Parkinson's disease and Huntington's chorea;
inflammatory autoimmune diseases such as multiple sclerosis, rheumatoid arthritis and Crohn's disease as well as other inflammatory disorders, such as cerebral ischemia , inflammatory bowel disease, ulcerative colitis, and atopic dermatitis; bone disease, such as osteoporosis, osteopetrosis, and Paget's disease; cancers, such as diffuse large-cell B cell lymphoma, chronic lymphocytic leukemia, acute lymphoblastic leukemia; Severe Acute Respiratory Syndrome;

and pre-term labor. Specific diseases or disorders which might be treated or prevented include those described herein, and in the references cited therein.

[0049] In certain einbodiments involving the compounds of the invention and the use thereof, the compound is specific for PDE4B, or an isoform thereof, e.g., PDE4B or PDE4D. In certain embodiments of aspects involving compounds of Fonnula I, Ia, Ib, Ic, Id, or Ie, the compound is specific for both PDE4B and PDE4D, specific for PDE4B, or specific for PDE4D. Such specificity means that the compound has at least 5-fold greater activity (preferably at least 5-, 10-, 20-, 50-, or 100-fold greater activity, or more) on PDE4B and/or PDE4D
than other enzymes, or on PDE4B relative to PDE4D, or PDE4D relative to PDE4B, where the activity is determined using a suitable assay, e.g., any assay known to one skilled in the art or as described herein.

[0050] In certain embodiments, a compound of the invention has an IC50 of less than 100 nM, less than 50 nM, less than 20 nM, less than 10 nM, less than 5 nM, or less than 1 nIVI with respect to at least one of PDE4B and PDE4D as determined in a generally accepted PDE4 activity assay. In one embodiment, a compound of Formula I, Ia, Ib, Ic, Id, or Ie will have an IC50 of less than 100 nM, less than 50 nM, less than 20 nM, less than 10 nM, less thasi 5 nM, or less than 1 nM with respect to PDE4B. In one embodiment, a compound of Formula I, Ia, Ib, Ic, Id, or Ie, will have an IC50 of less than 100 nM, less than 50 nM, less than 20 nM, less than nM, less than 5 nM, or less than 1 nM with respect to PDE4D. In one embodiment, a compound of Formula I, Ia, Ib, Ic, Id, or Ie will have an IC50 of less than 100 nM, less than 50 nM, less than 20 nM, less than 10 nM, less than 5 nM, or less than 1 nM with respect to both PDE4B and PDE4D. Further to any of the above embodiments, a compound of the invention will be a specific inhibitor of either PDE4B or PDE4D, such that the IC50 for one of PDE4B
and PDE4D will be at least about 5-fold, also 10-fold, also 20-fold, also 50-fold, or at least about 100-fold less than the IC50 for the other of PDE4B and PDE4D.

[0051] In certain embodiments of the invention, the compounds of Formula I, Ia, Ib, Ic, Id, or le, with activity on PDE4 will also have desireable pharmacologic properties.
In particular embodiments the desired pharmacologic property is any one or more of serum half-life longer than 2 hr (also longer than 4 hr, also longer than 8 hr), aqueous solubility, and oral bioavailability more than 10% (also more than 20%).

[0052] The identification of compounds of Formula I with activity on PDE4, or on isoforms thereof such as PDE4B or PDE4D, also provides a method for identifying or developing additional compounds with activity on PDE4 (or on the respective PDE4 isoform), e.g., improved modulators, by determining whether any of a plurality of test coinpounds of Formula I with activity on PDE4 provides an improvement in one or more desired pharmacologic properties relative to a reference compound with activity on PDE4, and selecting a compound if any, that has an improveinent in the desired pharmacologic property, thereby providing an improved modulator. In certain embodiments, the desired pharmacologic property is at least 2-fold, 4-fold, 6-fold, 8-fold, 10-fold, 20-fold, 50-fold, 100-fold, or more than 100-fold, greater activity on PDE4B than on PDE4D. In further embodiments, the desired pharmacologic property is an IC50 of less than 10 M, less than 1 M, less than 100 nM, less than 10 nM, or less than 1 nM.

[0053] In particular embodiments of the invention embracing modulator development, the desired pharmacologic property is serum half-life longer than 2 hr (or longer than 4 hr or longer than 8 hr), aqueous solubility, oral bioavailability more than 10% (or oral bioavailability more than 20%).

[0054] Also in particular embodiments of the invention embracing modulator development, the reference compound is a compound of Formula I. The process can be repeated multiple times, i.e., multiple rounds of preparation of derivatives and/or selection of additional related compounds and evaluation of such further derivatives of related compounds, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more additional rounds.

[0055] In additional aspects, structural information about PDE4B is utilized, e.g., in conjunction with compounds of Formula I or a molecular scaffold or scaffold core of Formula I. In addition, structural information about one or or more other PDEs can be utilized, e.g., PDE5A, PDE4D.

[0056] The invention also provides a method for developing ligands binding to a PDE4B, where the method includes identifying as molecular scaffolds one or more compounds that bind to a binding site of the PDE; determining the orientation of at least one molecular scaffold in co-crystals with the PDE; identifying chemical structures of one or more of the molecular scaffolds, that, when modified, alter the binding affinity or binding specificity or both between the molecular scaffold and the PDE; and synthesizing a ligand in which one or more of the chemical structures of the molecular scaffold is modified to provide a ligand that binds to the PDE with altered binding affinity or binding specificity or both. Such a scaffold can, for example, be a compound of Formula I, include a scaffold core as in Formula I, or include a bicyclic core as in Formula I.

[0057] In an alternative embodiment, the invention provides a method for developing or identifying ligands binding to a PDE4B. The method comprises deterinining the orientation of at least one molecular scaffold in co-crystals with the PDE wherein the molecular scaffold is contained with one or more compounds that bind to a binding site of the PDE;
identifying chemical structures of one or more of the molecular scaffolds, that, when modified, alter the binding affinity or binding specificity or both between the molecular scaffold and the PDE; and modifying the chemical structures so identified to provide a ligand that binds to the PDE with altered binding affinity or binding specificity or both. Such a scaffold can, for example, be a compound of Formula I, include a scaffold core as in Formula I, or include a bicyclic core as in Formula I.

[0058] In a related aspect, the invention provides a method for developing ligands specific for PDE4 (or a PDE4 isoform such as PDE4B or PDE4D), where the method involves determining whether a derivative of a compound that binds to a plurality of phosphodiesterases has greater specificity for the particular phosphodiesterase than the parent compound with respect to other phosphodiesterases. In one embodiment, the plurality of phosphodiesterases comprises PDE4B and PDE4D. In another embodiment, the plurality of phosphodiesterases comprises PDE4B and PDE5A. In a further embodiment, the compound that binds to a plurality of phosphodiesterases binds to PDE4B with an affinity at least 2-fold, e.g., 4-fold, 6-fold, 8-fold, 10-fold, 20-fold, 50-fold, or 100-fold, greater than for binding to any of the plurality of phosphodiesterases. In yet another embodiment, the compound that binds to a plurality of phosphodiesterases, binds weakly, wherein "weakly" indicates that the IC50 of the compound is greater than 1 M, e.g., 2 M, 5 M, 10 M, 20 M, 50 M, 100 M, 200 M, 500 M, 1 mM, or greater.

[0059] In another aspect, the invention provides a method for obtaining improved ligands binding to PDE4B, where the method involves identifying a compound that binds to that particular PDE, determining whether that compound interacts with one or more conserved active site residues, and determining whether a derivative of that compound binds to that PDE

with greater affinity or greater specificity or both than the parent binding compound. Binding with greater affinity or greater specificity or both than the parent compound indicates that the derivative is an improved ligand. This process can also be carried out in successive rounds of selection and derivatization and/or with multiple parent compounds to provide a compound or compounds with improved ligand characteristics. Likewise, the derivative compounds can be tested and selected to give high selectivity for that PDE, or to give cross-reactivity to a particular set of targets, for example to a subset of phosphodiesterases that includes PDE4B
and/or PDE4D. In particular embodiments, known PDE4B inhibitors can be used, and derivatives with greater affinity and/or greater specificity can be developed, preferably using PDE4B and/or PDE4D structure information; greater specificity for PDE4B
relative to PDE4D
is developed.

[0060] For Formula I. there are multiple scaffold cores described by Formula I. Such a scaffold core includes a bicyclic core as described above for Formula I, with an attached aryl or heteroaryl group of substituent group A as specified above. Thus, for each of the bicyclic cores shown above, there are corresponding scaffold cores for each of the positions of A, and for each aryl and heteroaryl group that is the initial moiety of A linked to the bicyclic core.
[0061] In another aspect, structural information about PDE4B can also be used to assist in determining a structure for another phosphodiesterase by creating a homology model from an electronic representation of a PDE4B structure. Such homology model is then equated with the structure of the other phosphodiesterase.

[0062] Typically, creating such a homology model involves identifying conserved amino acid residues between the known PDE having known structures, e.g., PDE4B, and the other phosphodiesterase of interest (e.g., PDE5A); transferring the atomic coordinates of a plurality of conserved amino acids in the known structure to the corresponding amino acids of the other phosphodiesterase to provide a rough structure of that phosphodiesterase; and constructing structures representing the remainder of the other phosphodiesterase using electronic representations of the structures of the remaining amino acid residues in the other phosphodiesterase. In particular, for PDE4B, coordinates from Table 1 and 2 can be used.
Conserved residues in a binding site can be used.

[0063] To assist in developing other portions of the phosphodiesterase structure, the homology model can also utilize, or be fitted with, low resolution x-ray diffraction data from one or more crystals of the phosphodiesterase, e.g., to assist in linking conserved residues and/or to better specify coordinates for terminal portions of a polypeptide.

[0064] The PDE4B structural infonnation used can be for a variety of different variants, including full-length wild type, naturally-occurring variants (e.g., allelic variants and splice variants), truncated variants of wild type or naturally-occuring variants, and mutants of full-length or truncated wild-type or naturally-occurring variants (that can be mutated at one or more sites). For example, in order to provide a PDE4B structure closer to a variety of other phosphodiesterase structures, a mutated PDE4B that includes a mutation to a conserved residue in a binding site can be used.

[0065] In another aspect, the invention provides a crystal comprising a crystalline form of PDE4B, which may be a reduced length PDE4B such as a phosphodiesterase domain, e.g., having atomic coordinates as described in Tables 1, 2, and 3. The crystalline form can contain one or more heavy metal atoms, for example, atoms useful for X-ray crystallography. The crystal can also include a binding compound in a co-crystal, e.g., a binding compound that interacts with one more more conserved active site residues in the PDE, or any two, any three, any four, any five, any six of those residues, and can, for example, be a known PDE inhibitor.
Such PDE crystals can be in various environments, e.g., in a crystallography plate, mounted for X-ray crystallography, and/or in an X-ray beam. The PDE may be of various forms, e.g., a wild-type, variant, truncated, and/or mutated form as described herein.

[0066] The invention further provides co-crystals of PDE4B, which may be a reduced length PDE, e.g., a phosphodiesterase domain, and a PDE4B binding compound. In certain embodiments, the binding compound within the crystal interacts with one or more conserved PDE4B active site residues. Advantageously, such co-crystals are of sufficient size and quality to allow structural determination of the PDE to at least 3 Angstroms, 2.5 Angstroms, 2.0 Angstroms, 1.8 Angstroms, 1.7 Angstroms, 1.5 Angstroms, 1.4 Angstroms, 1.3 Angstroms, or 1.2 Angstroms. The co-crystals can, for example, be in a crystallography plate, be mounted for X-ray crystallography and/or in an X-ray beam. Such co-crystals are beneficial, for example, for obtaining structural information concerning interaction between the PDE
and binding compounds.

[0067] In particular embodiments, the binding compound includes the bicyclic core or scaffold core structure as in Formula I, or is a compound of Formula I.

[0068] PDE4B binding compounds can include compounds that interact with at least one of conserved active site residues in the PDE, or any 2, 3, 4, 5, or 6 of those residues. Exemplary compounds that bind to PDE4B include compounds described in references cited herein.

[0069] Likewise, in additional aspects, methods for obtaining PDE4B crystals and co-crystals are provided. In one aspect, a method for obtaining a crystal of PDE4B
phosphodiesterase domain is provided by subjecting PDE4B protein at 5-20 mg/ml, e.g., 8-12 mg/ml, to crystallization conditions substantially equivalent to 30% PEG 400, 0.2M
MgC12, 0.1M Tris pH 8.5, 1 mM binding compound, at 4 C; or 20% PEG 3000, 0.2M Ca(OAc)a, 0.1M
Tris pH
7.0, 1 mM binding compound, 15.9 mg/ml protein at 4 C; or 1.8M -2.OM ammonium sulphate, 0.1 M CAPS pH 10.0 - 10.5, 0.2M lithium sulphate.

[0070] Crystallization conditions can be initially identified using a screening kit, such as a Hampton Research (Riverside, CA) screening kit 1. Conditions resulting in crystals can be selected and crystallization conditions optimized based on the demonstrated crystallization conditions. To assist in subsequent crystallography, the PDE can be seleno-methionine labeled. Also, as indicated above, the PDE may be any of various forms, e.g., truncated to provide a phosphodiesterase domain, which can be selected to be of various lengths.

[0071] In another aspect, the identification of compounds active on PDE4B
(such as compounds developed using methods described herein) makes it possible for one to modulate the PDE activity by contacting PDE with a compound that binds to PDE and interacts with one more conserved active site residues. The compound is preferably provided at a level sufficient to modulate the activity of the PDE by at least 10%, more preferably at least 20%, 30%, 40%, or 50%. In many embodiments, the compound will be at a concentration of about 1 M, 100 M, or 1 mM, or in a range of 1-100 nM, 100-500 nM, 500-1000 nM, 1-100 M, 100-500 M, or 500-1000 M.

[0072] In a related aspect, the invention provides a method for treating a subject suffering from a disease or condition characterized by abnormal PDE4 activity (e.g., abnormal PDE4B, PDE4D activity), where the method involves administering to the subject a compound identified by a method as described herein.

[0073] Because crystals of PDE4B have been developed and analyzed, and binding modes of ligands determined in such crystals, another aspect of the invention provides an electronic representation of these PDEs (which may be a reduced length PDE), for example, an electronic representation containing atomic coordinate representations for corresponding to the coordinates listed for PDE4B in Table 1 and 2, or a schematic representation such as one showing secondary structure and/or chain folding, and may also show conserved active site residues. The PDE may be wild type, an allelic variant, a mutant fonn, or a modifed form, e.g., as described herein. In particular, the PDE may consist essentially of a PDE4B phosphodiesterase domain.

[0074] The electronic representation can also be modified by replacing electronic representations of particular residues with electronic representations of other residues. Thus, for example, an electronic representation containing atomic coordinate representations corresponding to the coordinates for PDE4B listed in Tables 1 or 2 can be modified by the replacement of coordinates for a particular conserved residue in a binding site by a different amino acid. Following a modification or modifications, the representation of the overall structure can be adjusted to allow for the known interactions that would be affected by the modification or modifications. In most cases, a modification involving more than one residue will be performed in an iterative manner.

[0075] In addition, an electronic representation of a PDE4B binding compound or a test compound in the binding site can be included, e.g., a non-hydrolyzable cAMP
analog or a compound including the core structure of sildenafil.

[0076] Likewise, in a related aspect, the invention provides an electronic representation of a portion of PDE4B, which can be a binding site (which can be an active site) or phosphodiesterase domain, for example, PDE4B residues 152-528 of JC1519 (SEQ
ID NO:1), or other phosphodiesterase domain described herein. A binding site or phosphodiesterase domain can be represented in various ways, e.g., as representations of atomic coordinates of residues around the binding site and/or as a binding site surface contour, and can include representations of the binding character of particular residues at the binding site, e.g., conserved residues. The binding site preferably includes no more than 1 heavy metal atom; a binding compound or test compound such as a compound including the core structure of Formula I may be present in the binding site; the binding site may be of a wild type, variant, mutant form, or modified form of PDE4B; the electronic representation includes representations coordinates of conserved residues as in Table 1 or 2.

[0077] In yet another aspect, the structural and sequence information of PDE4B
can be used in a homology model for another PDE. It is helpful if high resolution structural information for PDE4B is used for such a model, e.g., at least 1.7, 1.5, 1.4, 1.3, or 1.2 Angstrom resolution.
[0078] In still another aspect, the invention provides an electronic representation of a modified PDE4B crystal structure, that includes an electronic representation of the atomic coordinates of a modified PDE4B based on the atomic coordinates of Table 1 and/or 2. In an exemplary embodiment, atomic coordinates of one of the listed tables can be modified by the replacement of atomic coordinates for a conserved residue with atomic coordinates for a different amino acid. Modifications can include substitutions, deletions (e.g., C-terminal and/or N-terminal delections), insertions (internal, C-terminal, and/or N-terminal) and/or side chain modifications.

[0079] In another aspect, the PDE4B structural information provides a method for developing useful biological agents based on PDE4B, by analyzing a PDE4B
structure to identify at least one sub-structure for forming the biological agent. Such sub-structures can include epitopes for antibody formation, and the method includes developing antibodies against the epitopes, e.g., by injecting an epitope presenting composition in a mammal such as a rabbit, guinea pig, pig, goat, or horse. The sub-structure can also include a mutation site at which mutation is expected to or is known to alter the activity of the PDE4B, and the method includes creating a mutation at that site. Still further, the sub-structure can include an attachment point for attaching a separate moiety, for example, a peptide, a polypeptide, a solid phase material (e.g., beads, gels, chromatographic media, slides, chips, plates, and well surfaces), a linker, and a label (e.g., a direct label such as a fluorophore or an indirect label, such as biotin or other member of a specific binding pair). The method can include attaching the separate moiety.

[0080] In another aspect, the invention provides a method for identifying potential PDE4B
binding compounds by fitting at least one electronic representation of a compound in an electronic representation of the PDE binding site. The representation of the binding site may be part of an electronic representation of a larger portion(s) or all of a PDE
molecule or may be a representation of only the catalytic domain or of the binding site or active site. The electronic representation may be as described above or otherwise described herein. For PDE4B the electronic representation includes representations of coordinates according to Tables 1 and/or 2 (in particular residues with coordinates differing signficantly from the previously proposed PDE4B structure). In certain embodiments, the compound complexed with PDE4B is an anlog of cGMP which is non-hydrolyzable.

[0081] In particular embodiments, the method involves fitting a computer representation of a compound from a computer database with a computer representation of the active site of the PDE, and involves removing a computer representation of a compound complexed with the PDE molecule and identifying compounds that best fit the active site based on favorable geometric fit and energetically favorable complementary interactions as potential binding compounds. In particular embodiments, the compound is a known PDE4B inhibitor, e.g., as described in a reference cited herein, or a derivative thereof.

[0082] In other embodiments, the method involves modifying a computer representation of a compound complexed with the PDE molecule, by the deletion or addition or both of one or more chemical groups; fitting a computer representation of a compound from a computer database with a computer representation of the active site of the PDE
molecule; and identifying compounds that best fit the active site based on favorable geometric fit and energetically favorable complementary interactions as potential binding compounds. In certain embodiments, the fitting comprises determining whether potential binding compounds interact with one or more of conserved PDE4B active site residues.

[0083] In still other embodiments, the method involves removing a computer representation of a compound complexed with the PDE, and searching a database for compounds having structural similarity to the complexed compound using a compound searching computer program or replacing portions of the complexed compound with similar chemical structures using a compound construction computer program.

[0084] Fitting a compound ca.n include determining whether a compound will interact with one or more conserved active site residues for the PDE. Compounds selected for fitting or that are complexed with the PDE can, for example, be a known PDE4B inhibitor compound, or a compound including the core structure of such compound.

[0085] In another aspect, the invention provides a method for attaching a PDE4B binding compound to an attachment component without substantially changing the binding of the binding compound to PDE4B, as well as a method for identifying attachment sites on a PDE4B

binding compound. The method involves identifying energetically allowed sites for attachment of an attachment component for the binding compoulld bound to a binding site of PDE4B; and attaching the compound or a derivative thereof to the attaclunent component at the energetically allowed site. In this context, "substantially changing the binding" denotes a change in IC50 after attachment of less than 3 log units, e.g., 1 nM to 1 M.
In certain embodiments, the binding compound is a compound of Formula I.

[0086] Attachment coinponents can include, for example, linkers (including traceless linkers) for attachment to a solid phase or to another molecule or other moiety. Such attachment can be formed by synthesizing the compound or derivative on the linker attached to a solid phase medium, e.g., in a combinatorial synthesis of a plurality of compounds.
Likewise, the attachinent to a solid phase medium can provide an affinity medium (e.g., for affinity chromatography).

[0087] The attachment component can also include a label, which can be a directly detectable label such as a fluorophore, or an indirectly detectable such as a member of a specific binding pair, e.g., biotin.

[0088] The ability to identify energentically allowed sites on a PDE4B binding compound, also, in a related aspect, provides modified binding compounds that have linkers attached, preferably at an energetically allowed site for binding of the modified compound to PDE4B.
The linker can be attached to an attachment component as described above.

[0089] Another aspect of the invention provides a modified PDE4B polypeptide that includes a modification that makes the modified PDE4B more similar than native PDE4B to another phosphodiesterase, and can also include other mutations or other modifications. In various embodiments, the polypeptide includes a full-length PDE4B polypeptide, includes a modified PDE4B binding site, includes at least 20, 30, 40, 50, 60, 70, or 80 contiguous amino acid residues derived from PDE4B including a conserved site.

[0090] Still another aspect of the invention provides a method for developing a ligand for a phosphodiesterase that includes conserved residues matching any one, 2, 3, 4, 5, or 6 of conserved PDE4B active site residues respectively, by determining whether a compound binds to the phosphodiesterase and interacts with such active site residues in a PDE4B crystal or a PDE4B binding model having coordinates as in Table 1 and/or 2. The method can also include determining whether the compound modulates the activity of the phosphodiesterase.
Preferably the phosphodiesterase has at least 50, 55, 60, or 70% identity over an equal length phosphodiesterase domain segment. In certain embodiments, the compound is a compound of Formula I.

[0091] In yet another aspect, the invention provides a method for developing or identifying a ligand for a phosphodiesterase wherein the phosphodiesterase comprises conserved residues matching one or more PDE4B active site residues. The method comprises determining whether a PDE4B binding compound binds to said phosphodiesterase, and determining whether the PDE4B binding compound interacts with one or more conserved PDE4B
active site residues in a crystal structure. In a further embodiment, the method comprises determining whether a PDE4B binding compound which binds to the phosphodiesterase interacts with one or more conserved PDE4B active site residues in a crystal structure.

[0092] In particular embodiments, determining includes computer fitting the compound in a binding site of the phosphodiesterase and/or the method includes fornning a co-crystal of the phosphodiesterase and the compound. Such co-crystals can be used for determining the binding orientation of the compound with the phosphodiesterase and/or provide structural information on the phosphodiesterase, e.g., on the binding site and interacting amino acid residues. Such binding orientation and/or other structural infonnation can be accomplished using X-ray crystallography.

[0093] The invention also provides compounds that bind to and/or modulate (e.g., inhibit) PDE4B phosphodiesterase activity e.g., compounds identified by the methods described herein.
Accordingly, in aspects of the invention involving PDE4B binding compounds, molecular scaffolds, and ligands or modulators, the compound is a weak binding compound;
a moderate binding compound; a strong binding compound; the compound interacts with one or more conserved active site residues in the PDE; the compound is a small molecule;
the compound binds to a plurality of different phosphodiesterases (e.g., at least 2, 3, 4, 5, 7, 10, or more different phosphodiesterases). In particular, the invention provides compounds identified or selected.

[0094] In yet another embodiment, the invention provides a method for identifying a compound having selectivity between PDE4B and PDE4D by utilizing particular differential sites. The method involves analyzing whether a compound differentially interacts in PDE4B

and PDE4D in at least one of the differential sites, where a differential interaction is indicative of such selectivity. The differential sites are identified from crystal structure comparison. The term "differential site" denotes a site, i.e., a location, where the chemical features of PDE4B and PDE4D interact differently with the compound. The term "chemical feature" is understood by those of skill in the art to denote structural and chemical properties responsible for chemical reactivity (including binding) and include without limitation hydrogen bond donor or acceptors, hydrophobic/lipophilic sites, positively ionizable sites, negatively ionizable sites, charge density, electronegativity, and the like.

[0095] In particular embodiments, the analyzing includes fitting an electronic representation of the compound in electronic representations of binding sites of PDE4B and PDE4D, and determining whether the compound differentially interacts based on said fitting; the method involves selecting an initial compound that binds to both PDE4B and PDE4D, fitting an electronic representation of the initial compound in electronic representations of binding sites of PDE4B and PDE4D, modifying the electronic representation of the initial compound with at least one moiety that interacts with at least differentials site, and determining whether the modified compound differentially binds to PDE4B and PDE4D; the modified compound binds differentially to a greater extent than does the initial compound; the method also includes assaying a compound that differentially interacts for differential activity on PDE4B and PDE4D; the initial compound includes the sildenafil scaffold structure; the initial compound can include the sildenafil core. Sildenafil is 1-[4-ethoxy-3-(6,7-dihydro-l-methyl-7-oxo-3-propyl-lH-pyrazolo[4,3-d]pyrimidin-5-yl)phenylsulfonyl]-4-methylpiperazine. In certain emboidments, the binding compound is a compound of Formula I.

[0096] In the various aspects described above that involve atomic coordinates for PDE4B in connection with binding compounds, the coordinates provided in Tables 1 or 2 can be used.
Those coordinates can then be adjusted using conventional modeling methods to fit compounds having structures different from sildenafil, and can thus be used for development of different PDE4B modulators, relative to currently described PDE4B modulators. PDE4B
crystal coordinates provided herein can be used instead of the previously described PDE4B structure coordinate because the present structure coordinates correct apparent errors in the previously described structure (e.g., as shown by the structure overlay in Figure 4), and thus are better adapted for PDE4B ligand development and other uses of PDE4B structure information such as the uses described herein.

[0097] Additional aspects and embodiments will be apparent from the following Detailed Description and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

[0098] FIGURE 1 shows a ribbon diagram schematic representation of PDE4B
phosphodiesterase domain having the sequence in Table 3.

[0099] FIGURE 2 shows an overlay of ribbon diagram schematic representations of the present PDE4B structure and the previously reported PDE4B crystal structure, and shows significant structure differences.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0100] Table 1 provides atomic coordinates for human PDE4B phosphodiesterase domain including residues 155-507. In this table, the various columns have the following content, beginning with the left-most column:
ATOM: Refers to the relevant moiety for the table row;
Atom number: Refers to the arbitrary atom number designation within the coordinate table;
Atom Name: Identifier for the atom present at the particular coordinates;
Chain ID: Chain ID refers to one monomer of the protein in the crystal, e.g., chain "A", or to other compound present in the crystal, e.g., HOH for water, and L
for a ligand or binding compound. Multiple copies of the protein monomers will have different chain Ids;
Residue Number: The amino acid residue number in the chain;
X, Y, Z: Respectively are the X, Y, and Z coordinate values;
Occupancy: Describes the fraction of time the atom is observed in the crystal.
For example, occupancy = 1 means that the atom is present all the time; occupancy =
0.5 indicates that the atom is present in the location 50% of the time;
B-factor: A measure of the thermal motion of the atom;
Element: Identifier for the element.

[0101] Table 2 provides atomic coordinate data for PDE4B phosphodiesterase domain co-crystal with 4-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 50.

[0102] Table 3 provides amino acid and nucleic acid sequences for PDE4B
phosphodiesterase domain as used in the work described herein.

[0103] Table 4 shows the alignment of the phosphodiesterase domains of PDE4B
and PDE4D, with 3 regions that can be exploited for designing selective ligands indicated by filled squares beneath the regions.

[0104] Table 5 provides activity of exemplary compounds of Formula I in assays that assess inhibition of PDE4B or PDE4D as described in Examples 145-147.

[0105] As used herein the following defmitions apply unless otherwise indicated:

[0106] "Halo" or "halogen" - alone or in combination means all halogens, that is, chloro (Cl), fluoro (F), bromo (Br), or iodo (I).

[0107] "Hydroxyl" refers to the group -OH.

[0108] "Thiol" or "mercapto" refers to the group -SH.

[0109] "Alkyl" - alone or in combination means an alkane-derived radical containing from 1 to 20, preferably 1 to 15, carbon atoms (unless specifically defined). It is a straight chain alkyl or branched alkyl, and includes a straight chain or branched alkyl group that contains or is interrupted by a cycloalkyl portion. The straight chain or branched alkyl group is attached at any available point to produce a stable compound. Examples of this include, but are not limited to, 4-(isopropyl)-cyclohexylethyl or 2-methyl-cyclopropylpentyl. In many einbodiments, an alkyl is a straight or branched alkyl group containing from 1-15, 1-8, 1-6, 1-4, or 1-2, carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, t-butyl and the like.
"Optionally substituted alkyl" denotes alkyl or alkyl that is independently substituted with 1 to 3 groups or substituents selected from the group consisting of halo, hydroxy, optionally substituted lower alkoxy, optionally substituted acyloxy, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted cycloalkyloxy, optionally substituted heterocycloalkyloxy, thiol, optionally substituted lower alkylthio, optionally substituted arylthio, optionally substituted heteroarylthio, optionally substituted cycloalkylthio, optionally substituted heterocycloalkylthio, optionally substituted alkylsulfinyl, optionally substituted arylsulfinyl, optionally substituted heteroarylsulfinyl, optionally substituted cycloalkylsulfinyl, optionally substituted heterocycloalkylsulfinyl, optionally substituted alkylsulfonyl, optionally substituted arylsulfonyl, optionally substituted heteroarylsulfonyl, optionally substituted cycloalkylsulfonyl, optionally substituted heterocycloalkylsulfonyl, optionally substituted amino, optionally substituted amido, optionally substituted amidino, optionally substituted urea, optionally substituted aminosulfonyl, optionally substituted alkylsulfonylamino, optionally substituted arylsulfonylamino, optionally substituted heteroarylsulfonylamino, optionally substituted cycloalkylsulfonylamino, optionally substituted heterocycloalkylsulfonylamino, optionally substituted alkylcarbonylamino, optionally substituted arylcarbonylamino, optionally substituted heteroarylcarbonylamino, optionally substituted cycloalkylcarbonylamino, optionally substituted heterocycloalkylcarbonylamino, optionally substituted carboxyl, optionally substituted acyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, nitro, and cyano, attached at any available point to produce a stable compound.

[0110] "Lower alkyl" refers to an alkyl group having 1-6 carbon atoms.
"Optionally substituted lower alkyl" denotes lower alkyl or lower alkyl that is independently substituted with 1 to 3 groups or substituents as defined in [0109] attached at any available point to produce a stable compound.

[0111] "Lower alkylene" refers to a divalent alkane-derived radical containing 1-6 carbon atoms, straight chain or branched, from which two hydrogen atoms are taken from the same carbon atom or from different carbon atoms. Examples of lower alkylene include, but are not limited to, -CH2-, -CH2CH2-, and -CH2CH(CH3)-.

[0112] "Alkenyl" - alone or in combination means a straight, branched, or cyclic hydrocarbon containing 2-20, preferably 2-17, more preferably 2-10, even more preferably 2-8, most preferably 2-4, carbon atoms and at least one, preferably 1-3, more preferably 1-2, most preferably one, carbon to carbon double bond. In the case of a cycloalkenyl group, conjugation of more than one carbon to carbon double bond is not such as to confer aromaticity to the ring.
Carbon to carbon double bonds may be either contained within a cycloalkyl portion, with the exception of cyclopropyl, or within a straight chain or branched portion.
Examples of alkenyl groups include ethenyl, propenyl, isopropenyl, butenyl, cyclohexenyl, cyclohexenylalkyl and the like. "Optionally substituted alkenyl" denotes alkenyl or alkenyl that is independently substituted with 1 to 3 groups or substituents as defined in [0109] attached at any available point to produce a stable compound.

[0113] "Lower alkenyl" refers to an alkenyl group having 1-6 carbon atoms.
"Optionally substituted lower alkenyl" denotes lower alkenyl or lower alkenyl that is substituted with 1 to 3 groups or substituents as defined in [0109] attached at any available point to produce a stable compound.

[0114] "Allcynyl" - alone or in combination means a straight or branched hydrocarbon containing 2-20, preferably 2-17, more preferably 2-10, even more preferably 2-8, most preferably 2-4, carbon atoms containing at least one, preferably one, carbon to carbon triple bond. Examples of alkynyl groups include ethynyl, propynyl, butynyl and the like.
"Optionally substituted alkynyl" denotes alkynyl or alkynyl that is independently substituted with 1 to 3 groups or substituents as defined in [0109] attached at any available point to produce a stable compound.

[0115] "Lower alkynyl" refers to an alkynyl group having 1-6 carbon atoms.
"Optionally substituted lower alkynyl" denotes lower alkynyl or lower alkynyl that is substituted with 1 to 3 groups or substituents as defined [0109] attached at any available point to produce a stable compound.

[0116] "Alkoxy" or "lower alkoxy" denotes the group -ORa, wherein Ra is alkyl or lower alkyl, respectively. "Optionally substituted alkoxy" or "optionally substituted lower alkoxy"
denotes alkoxy or lower alkoxy in which Ra is optionally substituted alkyl or optionally substituted lower alkyl, respectively.

[0117] "Acyloxy" denotes the group -OC(O)Rb, wherein Rb is hydrogen, lower alkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl. "Optionally substituted acyloxy" denotes acyloxy in which Rb is hydrogen, optionally substituted lower alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl.

[0118] "Aryloxy" denotes the group -OR , wherein R is aryl. "Optionally substituted aryloxy" denotes aryloxy or aryloxy in which R is optionally substituted aryl.

[0119] "Heteroaryloxy" denotes the group -ORd, wherein Rd is heteroaryl.
"Optionally substituted heteroaryloxy" denotes heteroaryloxy in which Rd is optionally substituted heteroaryl.

[0120] "Cycloalkyloxy" denotes the group -ORe, wherein Re is cycloalkyl.
"Optionally substituted cycloalkyloxy" denotes cycloalkyloxy in which Re is optionally substituted cycloalkyl.

[0121] "Heterocycloalkyloxy" denotes the group -OR; wherein Rf is heterocycloalkyl.
"Optionally substituted heterocycloalkyloxy" denotes heterocycloalkyloxy in which Rf is optionally substituted heterocycloalkyl.

[0122] "Alkylthio" or "lower alkylthio" denotes the group -OR, wherein Rg is alkyl or lower alkyl, respectively. "Optionally substituted alkylthio" or "optionally substituted lower alkylthio" denotes alkylthio or lower alkylthio in which Rg is optionally substituted alkyl or optionally substituted lower alkyl, respectively.

[0123] "Arylthio" denotes the group -SRh, wherein Rh is aryl. "Optionally substituted arylthio" denotes arylthio in which Rh is optionally substituted aryl.

[0124] "Heteroarylthio" denotes the group -S]R', wherein R' is heteroaryl.
"Optionally substituted heteroarylthio" denotes heteroarylthio in which R' is optionally substituted heteroaryl.

[0125] "Cycloalkylthio" denotes the group -SRj, wherein Ri is cycloalkyl.
"Optionally substituted cycloalkylthio" denotes cycloalkylthio in which Ri is optionally substituted cycloalkyl.

[0126] "Heterocycloalkylthio" denotes the group -SRk, wherein Rk is heterocycloalkyl.
"Optionally substituted heterocycloalkylthio" denotes heterocycloalkylthio in which Rk is optionally substituted heterocycloalkyl.

[0127] "Acyl" denotes groups -C(O)RL, wherein RL is hydrogen, lower alkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl. "Optionally substituted acyl" denotes acyl in which RL is hydrogen, optionally substituted lower alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl.

[0128] "Optionally substituted amino" denotes the group -NRmR", wherein R"' and R" are independently hydrogen, optionally substituted lower alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl,optionally substituted aryl, optionally substituted heteroaryl, optionally substituted acyl, or optionally substituted sulfonyl, or, Ri" and R"
together with the nitrogen to which they are attached form a 5-7 membered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring.

[0129] "Optionally substituted amido" denotes the group -C(O)NR Rp, wherein R
and Rp are independently hydrogen, optionally substituted lower alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl, or R and Rp together with the nitrogen to which they are attached form a 5-7 membered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring.

[0130] "Optionally substituted amidino" denotes the group -C(=NR4)NRrRs, wherein R4, Rr, and Rs are independently hydrogen or optionally substituted lower alkyl.

[0131] "Optionally substituted urea" denotes the group NRtC(O)NR R", wherein Rt is hydrogen or optionally substituted lower alkyl, and R and R are independently hydrogen, optionally substituted lower alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl or optionally substituted heteroaryl, or R and R'' together with the nitrogen to which they are attached form a 5-7 membered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring.

[0132] "Optionally substituted sulfonyl" denotes the group -S(O)2RW, wherein RW is optionally substituted lower alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl.

[0133] "Optionally substituted aminosulfonyl" denotes the group -S(O)ZNR"Ry, wherein R"
and Ry are independently hydrogen, optionally substituted lower alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl, or R" and R}' together with the nitrogen to which they are attached form a 5-7 membered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring.

[0134] "Carboxyl" denotes the group -C(O)ORZ, wherein RZ is hydrogen, lower alkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl. "Optionally substituted carboxyl" denotes carboxyl wherein RZ is hydrogen, optionally substituted lower alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl.

[0135] "Aryl" refers to a ring system-containing aromatic hydrocarbon such as phenyl or naphthyl, which may be optionally fused with a cycloalkyl of preferably 5-7, more preferably 5-6, ring members. "Optionally substituted aryl" denotes aryl or aryl that is substituted with 1 to 3 groups or substituents as defined in [0109], optionally substituted lower alkyl, optionally substituted lower alkenyl, or optionally substituted lower alkynyl, attached at any available point to produce a stable compound. A "substituted aryl" is aryl that is substituted with 1 to 3 groups or substituents as defined in [0109], or optionally substituted lower alkyl, optionally substituted lower alkenyl, or optionally substituted lower alkynyl, attached at any available point to produce a stable compound.

[0136] "Aralkyl" refers to the group -Raa-Ar wherein Ar is an aryl group and Raa is lower alkylene. "Optionally substituted aralkyl" denotes aralkyl or aralkyl in which the lower alkylene group is optionally substituted with 1 to 3 groups or substituents as defined in [0109]
attached at any available point to produce a stable compound, and in which the aryl group is optionally substituted with 1 to 3 groups or substituents as defmed in [0109], optionally substituted lower alkyl, optionally substituted lower alkenyl, or optionally substituted lower alkynyl, attached at any available point to produce a stable compound.

[0137] "Heteroaryl" alone or in combination refers to a monocyclic aromatic ring structure containing 5 or 6 ring atoms, or a bicyclic aromatic group having 8 to 10 atoms, containing one or more, preferably 1-4, more preferably 1-3, even more preferably 1-2, heteroatoms independently selected from the group consisting of 0, S, and N. Heteroaryl is also intended to include oxidized S or N, such as sulfinyl, sulfonyl, and N-oxide of a tertiary ring nitrogen.
A carbon or nitrogen atom is the point of attachment of the heteroaryl ring structure such that a stable aromatic ring is retained. Examples of heteroaryl groups include, but are not limited to, pyridinyl, pyridazinyl, pyrazinyl, quinaoxalyl, indolizinyl, benzo[b]thienyl, quinazolinyl, purinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl, oxazolyl, thiazolyl, thienyl, isoxazolyl, oxathiadiazolyl, isothiazolyl, tetrazolyl, imidazolyl, triazinyl, furanyl, benzofuryl, and indolyl.

"Optionally substituted heteroaryl" includes heteroaryl or heteroaryl that is substituted with 1 to 3 groups or substituents as defined in [0109], optionally substituted lower alkyl, optionally substituted lower alkenyl, or optionally substituted lower alkynyl, attached at any available carbon or nitrogen to produce a stable compound. "Substituted heteroaryl"
denotes heteroaryl that is substituted with 1 to 3 groups or substituents as defined in [01091, or optionally substituted lower alkyl, optionally substituted lower alkenyl, or optionally substituted lower alkynyl, attached at any available carbon or nitrogen to produce a stable compound [0138] "Heteroaralkyl" refers to the group -Rbb-HetAr wherein HetAr is a heteroaryl group, and Rbb is lower alkylene. "Optionally substituted heteroaralkyl" denotes heteroaralkyl or heteroaralkyl in which the lower alkylene group is optionally substituted with 1 to 3 groups or substituents as defined in [0109], attached at any available point to produce a stable compound, and in which the heteroaryl group is optionally substituted with 1 to 3 groups or substituents as defined in [0109], optionally substituted lower alkyl, optionally substituted lower alkenyl, or optionally substituted lower alkynyl, attached at any available point to produce a stable compound.

[0139] "Cycloalkyl" refers to saturated or unsaturated, non-aromatic monocyclic, bicyclic or tricyclic carbon ring systems of 3-8, more preferably 3-6, ring members per ring, such as cyclopropyl, cyclopentyl, cyclohexyl, adamantyl, and the like. "Optionally substituted cycloalkyl" denotes cycloalkyl or cycloalkyl that is substituted with 1 to 3 groups or substituents as defined in [0109], optionally substituted lower alkyl, optionally substituted lower alkenyl, or optionally substituted lower alkynyl, attached at any available carbon or nitrogen to produce a stable compound.

[0140] "Cycloalkylalkyl" refers to the group -R -Cyc wherein Cyc is a cycloalkyl group, and R ' is a lower alkylene group. "Optionally substituted cycloalkylalkyl"
denotes cycloalkylalkyl or cycloalkylalkyl in which the lower alkylene group is optionally substituted with 1 to 3 groups or substituents as defined in [0109], attached at any available point to produce a stable compound, and in which the cycloalkyl group is optionally substituted with 1 to 3 groups or substituents as defined in [0109], optionally substituted lower alkyl, optionally substituted lower alkenyl, or optionally substituted lower alkynyl, attached at any available point to produce a stable compound.

[0141] "Heterocycloalkyl" refers to a saturated or unsaturated non-aromatic cycloalkyl group having from 5 to 10 atoms in which from 1 to 3 carbon atoms in the ring are replaced by heteroatoms of 0, S or N, and are optionally fused with benzo or heteroaryl of 5-6 ring members. Heterocycloalkyl is also intended to include oxidized S or N, such as sulfinyl, sulfonyl and N-oxide of a tertiary ring nitrogen. The point of attachment of the heterocycloalkyl ring is at a carbon or nitrogen atom such that a stable ring is retained.
Examples of heterocycloalkyl groups include, but are not limited to, morpholino, tetrahydrofiiranyl, dihydropyridinyl, piperidinyl, pyrrolidinyl, piperazinyl, dihydrobenzofuryl, and dihydroindolyl. "Optionally substituted heterocycloalkyl" denotes heterocycloalkyl or heterocycloalkyl that is substituted with 1 to 3 groups or substituents as defined in [0109], optionally substituted lower alkyl, optionally substituted lower alkenyl, or optionally substituted lower alkynyl, attached at any available carbon or nitrogen to produce a stable compound.

[0142] "Heterocycloalkylalkyl" refers to the group -Rdd-Het wherein Het is a heterocycloalkyl group, and Rad is a lower alkylene group. "Optionally substituted heterocycloalkylalkyl" denotes heterocycloalkylalkyl or heterocycloalkylalkyl in which the lower alkylene group is optionally substituted with 1 to 3 groups or substituents as defined in [0109], attached at any available point to produce a stable compound, and in which the heterocycloalkyl group is optionally substituted with 1 to 3 groups or substituents as defined in [0109], optionally substituted lower alkyl, optionally substituted lower alkenyl, or optionally substituted lower alkynyl, attached at any available point to produce a stable coinpound.

[0143] "Optionally substituted alkylsulfinyl" denotes the group -S(O)Ree, wherein Ree is optionally substituted lower alkyl.

[0144] "Optionally substituted arylsulfinyl" denotes the group -S(O)Rf ;
wherein Rff is optionally substituted aryl.

[0145] "Optionally substituted heteroarylsulfinyl" denotes the group -S(O)Rgg, wherein Rgg is optionally substituted heteroaryl.

[0146] "Optionally substituted cycloalkylsulfinyl" denotes the group -S(O)Rhh, wherein Rhl' is optionally substituted cycloalkyl.

[0147] "Optionally substituted heterocycloalkylsulfinyl" denotes the group -S(O)R", wherein R" is optionally substituted heterocycloalkyl.

[0148] "Optionally substituted alkylsulfonyl" denotes the group -S(O)2Rj, wherein Rj is optionally substituted lower alkyl.

[0149] "Optionally substituted arylsulfonyl" denotes the group -S(O)2Rkk, wherein Rkk is optionally substituted aryl.

[0150] "Optionally substituted heteroarylsulfonyl" denotes the group -S(O)2RLL, wherein RLL is optionally substituted heteroaryl.

[0151] "Optionally substituted cycloalkylsulfonyl" denotes the group -S(O)aR""", wherein R7 is optionally substituted cycloalkyl.

[0152] "Optionally substituted heterocycloalkylsulfonyl" denotes the group -S(O)2R, wherein R"" is optionally substituted heterocycloalkyl.

[0153] "Optionally substituted alkylsulfonylamino" denotes the group =NR
S(O)2Rpp, wherein Rpp is optionally substituted lower alkyl, and R00 is hydrogen or optionally substituted lower alkyl.

[0154] "Optionally substituted arylsulfonylamino" denotes the group NR
S(O)2Rqq, wherein Raq is optionally substituted aryl, and R is liydrogen or optionally substituted lower alkyl.

[0155] "Optionally substituted heteroarylsulfonylamino" denotes the group -NROOS(O)2R7, wherein R'T is optionally substituted heteroaryl, and RO0 is hydrogen or optionally substituted lower alkyl.

[0156] "Optionally substituted cycloalkylsulfonylamino" denotes the group -NR
S(O)2RS5, wherein Rss is optionally substituted cycloalkyl, and R is hydrogen or optionally substituted lower alkyl.

[0157] "Optionally substituted heterocycloalkylsulfonylamino" denotes the group -NR S(O)zRtt, wherein Rtt is optionally substituted heterocycloalkyl, and R
is hydrogen or optionally substituted lower alkyl.

[0158] "Optionally substituted alkylcarbonylamino" denotes the group NR C(O)R
II, wherein R is optionally substituted lower alkyl, and R0 is hydrogen or optionally substituted lower alkyl.

[0159] "Optionally substituted arylcarbonylamino" denotes the group NR
C(O)R''", wherein R"'' is optionally substituted aryl, and RO0 is hydrogen or optionally substituted lower alkyl.

[0160] "Optionally substituted heteroarylcarbonylamino" denotes the group -NR
0C(O)R7, wherein Rw' is optionally substituted heteroaryl, and R is hydrogen or optionally substituted lower alkyl.

[0161] "Optionally substituted cycloalkylcarbonylamino" denotes the group -NR
C(O)RW', wherein Re' is optionally substituted cycloalkyl, and R is hydrogen or optionally substituted lower alkyl.

[0162] "Optionally substituted heterocycloalkylcarbonylamind" denotes the group -NR00C(O)Ryy, wherein Ry}' is optionally substituted heterocycloalkyl, and R
is hydrogen or optionally substituted lower alkyl.

[0163] As used herein, the terms "composition" and "pharmaceutical composition" refer to a preparation that includes a therapeutically significant quantity of an active agent, that is prepared in a form adapted for administration to a subject. Thus, the preparation does not include any component or components in such quantity that a reasonably prudent medical practitioner would find the preparation unsuitable for administration to a normal subject. In many cases, such a pharmaceutical composition is a sterile preparation.

[0164] As used herein in connection with PDE4 modulating compound, binding compounds or ligands, the term "specific for PDE4 phosphodiesterase", "specific for PDE4" and terms of like import mean that a particular compound binds to PDE4 to a statistically greater extent than to other phosphodiesterases that may be present in a particular organism, e.g., at least 2, 3, 4, 5, 10, 20, 50, 100, or 1000-fold. Also, where biological activity other than binding is indicated, the term "specific for PDE4" indicates that a particular compound has greater biological activity associated with binding PDE4 than to other phosphodiesterases (e.g., at a level as indicated for binding specificity). Preferably, the specificity is also with respect to other biomolecules (not limited to phosphodiesterases) that may be present from an organism. Such binding and/or activity specificity may be for a PDE4 isoform, e.g., PDE4A, PDE4B, PDE4C, PDE4D, such that the specificity is also with respect to the other PDE4 isoforms. In the context of ligands interacting with PDE4, the terms "activity on", "activity toward," and like terms mean that such ligands have IC50 less than 10 M, less than 1 M, less than 100 nM, less than 50 nM, less than 20 nM, less than 10 nM, less than 5 nM, or less than 1 nM with respect to one or more PDE4 as determined in a generally accepted PDE4 activity assay.

[0165] As used herein, the tei7n "PDE4-mediated" disease or condition and like terms refer to a disease or condition in wliich the biological function of PDE4 affects the development and/or course of the disease or condition, and/or in which modulation of PDE4 alters the development, course, and/or symptoms of the disease or condition. Similarly, the phrase "PDE4 modulation provides a therapeutic benefit" indicates that modulation of the level of activity of PDE4 in a subject indicates that such modulation reduces the severity and/or duration of the disease, reduces the likelihood or delays the onset of the disease or condition, and/or causes an improvement in one or more symptoms of the disease or condtion. In some cases the disease or condition may be mediated by one of the the PDE4 isoforms, e.g., PDE4B, PDE4C, or PDE4D.

[0166] In the present context, the term "therapeutically effective" indicates that the materials or amount of material is effective to prevent, alleviate, or ameliorate one or more symptoms of a disease or medical condition, and/or to prolong the survival of the subject being treated.

[0167] The term "pharmaceutically acceptable" indicates that the indicated material does not have properties that would cause a reasonably prudent medical practitioner to avoid administration of the material to a patient, taking into consideration the disease or conditions to be treated and the respective route of administration. For example, it is commonly required that such a material be essentially sterile, e.g., for injectibles.

[0168] The term "pharmaceutically acceptable metabolite" refers to a pharmacologically acceptable product, which may be an active product, produced through metabolism of a specified compound (or salt thereof) in the body of a subject or patient.
Metabolites of a compound may be identified using routine techniques known in the art, and their activities determined using tests such as those described herein. For example, in some compounds, one or more alkoxy groups can be metabolized to hydroxyl groups while retaining pharmacologic activity and/or carboxyl groups can be esterified, e.g., glucuronidation. In some cases, there can be more than one metabolite, where an intermediate metabolite(s) is further metabolized to provide an active metabolite. For example, in some cases a derivative compound resulting from metabolic glucuronidation may be inactive or of low activity, and can be further metabolized to provide an active metabolite.

[0169] "A pharmaceutically acceptable salt" is intended to mean a salt that retains the biological effectiveness of the free acids and bases of the specified compound and that is not biologically or otherwise unacceptable. A compound of the invention may possess a sufficiently acidic, a sufficiently basic, or both functional groups, and accordingly react with any of a number of inorganic or organic bases, and inorganic and organic acids, to form a pharmaceutically acceptable salt. Exemplary pharmaceutically acceptable salts include those salts prepared by reaction of the compounds of the present invention with a mineral or organic acid or an inorganic base, such as salts including sodium, chloride, sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyne-1,4 dioates, hexyne-1,6-dioates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phtlialates, sulfonates, xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, .gamma.-hydroxybutyrates, glycollates, tartrates, methane-sulfonates, propanesulfonates, naphthalene-l-sulfonates, naphthalene-2-sulfonates, and mandelates.

[0170] The terms "PDE4B phosphodiesterase" and "PDE4B" mean an enzymatically active phosphodiesterase that contains a portion with greater than 90% amino acid sequence identity to amino acid residues 152-528 (S152-S528) with reference to GenBank polypeptide sequence JC1519 (SEQ ID NO:1) of native PDE4B as shown in Table 4, for a maximal alignment over an equal length segment; or that contains a portion with greater than 90%
amino acid sequence identity to at least 200 contiguous amino acids from amino acid residues 152-528 of JC1519 (SEQ ID NO:1) of native PDE4B that retains binding to natural PDE4B ligand.
Preferably the sequence identity is at least 95, 97, 98, 99, or even 100%. Preferably the specified level of sequence identity is over a sequence at least 300 contiguous amino acid residues in length.
The sequence represented by amino acid residues 152-528 of JC1519 (SEQ ID
NO:1) is also available as S324 to S700 of NP 002591 (SEQ ID NO:3, encoded by NM_002600, SEQ
ID

NO:4), S309 to S685 of AAB96381 (SEQ ID NO:5), and S194 to S570 of AAA35643 (SEQ
ID NO:6). Therefore, amino acid residues identified in one of the listed sequences can also be expressed as the matching amino acid residue in any other of the listed sequences or other matching sequence.

[0171] The term "PDE4B phosphodiesterase domain" refers to a reduced length PDE4B (i.e., shorter than a full-length PDE4B by at least 100 amino acids that includes the phosphodiesterase catalytic region in PDE4B. Highly preferably for use in this invention, the phosphodiesterase domain retains phosphodiesterase activity, preferably at least 50% the level of phosphodiesterase activity as compared to the native PDE4B, more preferably at least 60, 70, 80, 90, or 100% of the native activity.

[0172] As used herein, the terms "ligand" and "modulator" are used equivalently to refer to a compound that modulates the activity of a target biomolecule, e.g., an enzyme such as a kinase or phosphodiesterase. Generally a ligand or modulator will be a small molecule, where "small molecule refers to a compound with a molecular weight of 1500 daltons or less, or preferably 1000 daltons or less, 800 daltons or less, or 600 daltons or less.
Thus, an "iinproved ligand" is one that possesses better pharmacological and/or pharmacokinetic properties than a reference compound, where "better" can be defined by a person for a particular biological system or therapeutic use. In terms of the development of ligands from scaffolds, a ligand is a derivative of a scaffold.

[0173] In the context of bindiuig compounds, molecular scaffolds, and ligands, the term "derivative" or "derivative compound" refers to a compound having a chemical structure that contains a common core chemical structure as a parent or reference compound, but differs by having at least one structural difference, e.g., by having one or more substituents added and/or removed and/or substituted, and/or by having one or more atoms substituted with different atoms. Unless clearly indicated to the contrary, the term "derivative" does not mean that the derivative is synthesized using the parent compound as a starting material or as an intermediate, although in some cases, the derivative may be synthesized from the parent.
[0174] Thus, the term "parent compound" refers to a reference compound for another compound, having structural features continued in the derivative compound.
Often but not always, a parent compound has a simpler chemical stracture than the derivative.

[0175] By "chemical structure" or "chemical substructure" is meant any definable atom or group of atoms that constitute a part of a molecule. Normally, chemical substructures of a scaffold or ligand can have a role in binding of the scaffold or ligand to a target molecule, or can influence the three-dimensional shape, electrostatic charge, and/or conformational properties of the scaffold or ligand.

[0176] The term "binds" in connection with the interaction between a target and a potential binding compound indicates that the potential binding compound associates with the target to a statistically significant degree as compared to association with proteins generally (i.e., non-specific binding). Thus, the term "binding compound" refers to a compound that has a statistically significant association with a target molecule. Preferably a binding compound interacts with a specified target with a dissociation constant (kd) of 1 mM or less. A binding compound can bind with "low affinity", "very low affinity", "extremely low affinity", "moderate affinity", "moderately high affinity", or "high affinity" as described herein.

[0177] In the context of compounds binding to a target, the term "greater affinity" indicates that the compound binds more tightly than a reference compound, or than the saine compound in a reference condition, i.e., with a lower dissociation constant. In particular embodiments, the greater affinity is at least 2, 3, 4, 5, 8, 10, 50, 100, 200, 400, 500, 1000, or 10,000-fold greater affinity.

[0178] Also in the context of compounds binding to a biomolecular target, the term "greater specificity" indicates that a compound binds to a specified target to a greater extent than to another biomolecule or biomolecules that may be present under relevant binding conditions, where binding to such other biomolecules produces a different biological activity than binding to the specified target. Typically, the specificity is with reference to a limited set of other biomolecules, e.g., in the case of PDE4B, other phosphodiesterases (e.g., PDE5A) or other PDE4 isoforms (e.g., PDE4D) or even other type of enzymes. In particular embodiments, the greater specificity is at least 2, 3, 4, 5, 8, 10, 50, 100, 200, 400, 500, or 1000-fold greater specificity.

[0179] As used in connection with binding of a compound with a target, the term "interact"
indicates that the distance from a bound compound to a particular amino acid residue will be 5.0 angstroms or less. In particular embodiments, the distance from the compound to the particular amino acid residue is 4.5 angstroms or less, 4.0 angstroms or less, or 3.5 angstroms or less. Such distances can be determined, for example, using co-crystallography, or estimated using computer fitting of a compound in an active site.

[0180] Reference to particular amino acid residues in PDE4B polypeptide residue number is defined by the numbering corresponding to NCBI protein sequence accession number JC1519 (SEQ ID NO: 1), as described, for example, in McLaughlin et al., J. Biol.
Chem. 268 (9), 6470-6476 (1993); Obernolte et al., Gene 129 (2), 239-247 (1993); and Bolger et al., Mol. Cell. Biol.
13 (10), 6558-6571 (1993). As indicated above, alternate numbering from other matching PDE4B sequences can also be used.

[0181] By "molecular scaffold" or "scaffold" is meant a simple target binding molecule to which one or more additional chemical moieties can be covalently attached, modified, or eliminated to form a plurality of molecules with corninon structural elements.
The moieties can include, but are not limited to, a halogen atom, a hydroxyl group, a methyl group, a nitro group, a carboxyl group, or any other type of molecular group including, but not limited to, those recited in this application. Molecular scaffolds bind to at least one target molecule, preferably to a plurality of molecules in a protein family, and the target molecule can preferably be a enzyme, receptor, or other protein. Preferred characteristics of a scaffold can include binding at a target molecule binding site such that one or more substituents on the scaffold are situated in binding pockets in the target molecule binding site;
having chemically tractable structures that can be chemically modified, particularly by synthetic reactions, so that a combinatorial library can be easily constructed; having chemical positions where moieties can be attached that do not interfere with binding of the scaffold to a protein binding site, such that the scaffold or libraay members can be modified to form ligands, to achieve additional desirable characteristics, e.g., enabling the ligand to be actively transported into cells and/or to specific organs, or enabling the ligaazd to be attached to a chromatography column for additional analysis. Thus, a molecular scaffold is an identified target binding molecule prior to modification to improve binding affinity and/or specificity, or other pharmacalogic properties.
[0182] The term "scaffold core" refers to the core structure of a molecular scaffold onto which various substituents can be attached. Thus, for a number of scaffold molecules of a particular chemical class, the scaffold core is common to all the scaffold molecules. In many cases, the scaffold core will consist of or include one or more ring structures.

[0183] By "binding site" is meant an area of a target molecule to which a ligand can bind non-covalently. Binding sites embody particular shapes and often contain multiple binding pockets present within the binding site. The particular shapes are often conserved within a class of molecules, such as a molecular family. Binding sites within a class also can contain conserved structures such as, for example, chemical moieties, the presence of a binding pocket, and/or an electrostatic charge at the binding site or some portion of the binding site, all of which can influence the shape of the binding site.

[0184] By "binding pocket" is meant a specific volume within a binding site. A
binding pocket can often be a particular shape, indentation, or cavity in the binding site. Binding pockets can contain particular chemical groups or structures that are important in the non-covalent binding of another molecule such as, for example, groups that contribute to ionic, hydrogen bonding, or van der Waals interactions between the molecules.

[0185] By "orientation", in reference to a binding compound bound to a target molecule is meant the spatial relationship of the binding compound (which can be defined by reference to at least some of its consitituent atoms) to the binding pocket and/or atoms of the target molecule at least partially defining the binding pocket.

[0186] In the context of target molecules in this invention, the term "crystal" refers to a regular assemblage of a target molecule of a type suitable for X-ray crystallography. That is, the assemblage produces an X-ray diffraction pattern when illuminated with a beam of X-rays.
Thus, a crystal is distinguished from an aggolmeration or other complex of target molecule that does not give a diffraction pattern.

[0187] By "co-crystal" is meant a complex of the compound, molecular scaffold, or ligand bound non-covalently to the target molecule and present in a crystal form appropriate for analysis by X-ray or protein crystallography. In preferred embodiments the target molecule-ligand complex can be a protein-ligand complex.

[0188] The phrase "alter the binding affinity or binding specificity" refers to changing the binding constant of a first compound for another, or changing the level of binding of a first compound for a second compound as compared to the level of binding of the first compound for third compounds, respectively. For example, the binding specificity of a compound for a particular protein is increased if the relative level of binding to that particular protein is increased as compared to binding of the compound to unrelated proteins.

[0189] As used herein in connection with test compounds, binding compounds, and modulators (ligands), the term "synthesizing" and like terms means chemical synthesis from one or more precursor materials.

[0190] The phrase "chemical structure of the molecular scaffold is modified"
means that a derivative molecule has a chemical structure that differs from that of the molecular scaffold but still contains common core chemical structural features. The phrase does not necessarily mean that the molecular scaffold is used as a precursor in the synthesis of the derivative.

[0191] By "assaying" is meant the creation of experimental conditions and the gathering of data regarding a particular result of the experimental conditions. For example, enzymes can be assayed based on their ability to act upon a detectable substrate. A compound or ligand can be assayed based on its ability to bind to a particular target molecule or molecules.

[0192] By a "set" of compounds is meant a collection of compounds. The compounds may or may not be structurally related.

[0193] As used herein, the term "modulating" or "modulate" refers to an effect of altering a biological activity, especially a biological activity associated with a particular biomolecule such as PDE4 or an isoform thereof, e.g., PDE4B. For example, an agonist or antagonist of a particular biomolecule modulates the activity of that biomolecule, e.g., an enzyme.

[0194] The term "PDE4 activity" refers to a biological activity of PDE4, particularly including phosphodiesterase activity. Similar terms apply to the particular PDE4 isoforms, e.g., PDE4A, PDE4B, PDE4C, and PDE4D.

[0195] In the context of the use, testing, or screening of compounds that are or may be modulators, the term "contacting" means that the compound(s) are caused to be in sufficient proximity to a particular molecule, complex, cell, tissue, organism, or other specified material that potential binding interactions andlor chemical reaction between the compound and other specified material can occur.

1. General [0196] The present invention provides coinpounds of Formula that are inhibitors of PDE4B, and methods for the use of PDE4B phosphodiesterase structures, structural information, and related compositions for developing improved compounds with those structures that modulate PDE4B phosphodiesterase activity.

[0197] A number of patent publications have concerned PDE4 inhibitors and their use. Most such publications have focused on PDE4D. For example, Marfat et al., U.S.
Patent 6,559,168 describes PDE4 inhibitors, especially PDE4D inhibitors, and cites additional patent publications that describe additional PDE4 inhibitors. Such additional publications include Marfat et al., WO 98/45268; Saccoomano et al., U.S. Patent 4,861,891; Pon, U.S. Patent 5,922,557; and Eggleston, WO 99/20625.

[0198] Ait Ikhlef et al., U.S. Patent Publ. 20030064374, Appl. No. 10/983,754 describes compounds active on PDE4B and their use in treatment of neurotoxicity, including treatment in neurodegenerative diseases such as Alzheimers' disease, Parkinson's disease, multiple sclerosis, Huntington's chorea, and cerebral ischemia.

[0199] All of the cited references above are incorporated herein by reference in their entireties, including without limitation for the descriptions of inhibitors and their uses as well as for assays, syntheses, and for identification and preparation of the PDEs and derivatives.

Exemplary Diseases Associated with PDE4B.

[02001 Modulation of PDE4B has been correlated with treatment of a number of different diseases and conditions. A number of patent publications have described PDE4 inhibitors and their use. Most such publications have focused on PDE4D. For example, Marfat et al., U.S.
Patent 6,559,168 describes PDE4 inhibitors, especially PDE4D inhibitors, and cites additional patent publications that describe additional PDE4 inhibitors. Such additional publications include Marfat et al., WO 98/45268; Saccoomano et al., U.S. Patent 4,861,891;
Pon, U.S.
Patent 5,922,557; and Eggleston, WO 99/20625.

[0201] Ait Ikhlef et al., U.S. Patent Publ. 20030064374, Appl. No. 10/983,754 describes compounds active on PDE4B and their use in treatment of neurotoxicity, including treatment in neurodegenerative diseases such as Alzheimers' disease, Parkinson's disease, multiple sclerosis, Huntington's chorea, and cerebral ischemia.

[0202] Thus, PDE4B modulators can be used for treatement or prophylaxis of such conditions correlated with PDE4 and in particular PDE4B. Additional conditions that can be treated include, without limitation, an acute or chronic pulmonary disease such as obstructive diseases (e.g. asthma, chronic obstructive pulmonary disease (COPD), cystic fibrosis), interstitial lung diseases (e.g. idiopathic pulmonary fibrosis, sarcoidosis), vascular lung diseases (e.g. pulmonary hypertension), bronchitis, allergic bronchitis, and emphysema.
Additional diseases or conditions contemplated for treatment by embodiments of the present invention include for example, without limitation, CNS diseases such as Alzheimer's disease, Parkinson's disease and Huntington's chorea; inflaminatory autoimmune diseases such as multiple sclerosis, rheumatoid arthritis and Crohn's disease as well as other inflammatory disorders, such as cerebral ischemia, inflammatory bowel disease, ulcerative colitis, and atopic dermatitis; bone disease, such as osteoporosis, osteopetrosis, and Paget's disease; cancers, such as diffuse large-cell B cell lymphoma, chronic lyinphocytic leukemia, acute lymphoblastic leukeinia; Severe Acute Respiratory Syndrome; and pre-term labor.

II. Crystalline PDE4B

[0203] Crystalline PDE4B includes native crystals, phosphodiesterase domain crystals, derivative crystals and co-crystals. The native crystals generally comprise substantially pure polypeptides corresponding to PDE4B in crystalline form. PDE4B
phosphodiesterase domain crystals generally comprise substantially pure PDE4B phosphodiesterase domain in crystalline form. In connection with the development of inhibitors of PDE4B
phosphodiesterase function, it is advantageous to use PDE4B phosphodiesterase domain respectively for structural determination, because use of the reduced sequence simplifies structure determination. To be useful for this purpose, the phosphodiesterase domain should be active and/or retain native-type binding, thus indicating that the phosphodiesterase domain takes on substantially normal 3D structure.

[0204] It is to be understood that the crystalline phosphodiesterases and phosphodiesterase domains of the invention are not limited to naturally occurring or native phosphodiesterase.
Indeed, the crystals of the invention include crystals of mutants of native phosphodiesterases.
Mutants of native phosphodiesterases are obtained by replacing at least one amino acid residue in a native phosphodiesterase with a different amino acid residue, or by adding or deleting amino acid residues within the native polypeptide or at the N- or C-terminus of the native polypeptide, and have substantially the same three-dimensional structure as the native phosphodiesterase from which the mutant is derived.

[0205] By having substantially the same three-dimensional structure is meant having a set of atomic structure coordinates that have a root-mean-square deviation of less than or equal to about 2A when superimposed with the atomic structure coordinates of the native phosphodiesterase from which the mutant is derived when at least about 50% to 100% of the Ca atoms of the native phosphodiesterase domain are included in the superposition.

[0206] Amino acid substitutions, deletions and additions which do not significantly interfere with the three-dimensional structure of the phosphodiesterase will depend, in part, on the region of the phosphodiesterase where the substitution, addition or deletion occurs. In highly variable regions of the molecule, non-conservative substitutions as well as conservative substitutions may be tolerated without significantly disrupting the three-dimensional, structure of the molecule. In highly conserved regions, or regions containing significant secondary structure, conservative amino acid substitutions are preferred. Such conserved and variable regions can be identified by sequence alignment of PDE4B with other phosphodiesterases.
[0207] Conservative amino acid substitutions are well known in the art, and include substitutions made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity and/or the amphipathic nature of the amino acid residues involved. For example, negatively charged amino acids include aspartic acid and glutamic acid; positively charged amino acids include lysine and arginine; amino acids with uncharged polar head groups having similar hydrophilicity values include the following: leucine, isoleucine, valine;
glycine, alanine; asparagine, glutamine; serine, threonine; phenylalanine, tyrosine. Other conservative amino acid substitutions are well known in the art.

[0208] For phosphodiesterases obtained in whole or in part by chemical synthesis, the selection of amino acids available for substitution or addition is not limited to the genetically encoded amino acids. Indeed, the mutants described herein may contain non-genetically encoded amino acids. Conservative amino acid substitutions for many of the commonly known non-genetically encoded amino acids are well known in the art.
Conservative substitutions for other amino acids can be detezmined based on their physical properties as compared to the properties of the genetically encoded amino acids.

[0209] In some instances, it may be particularly advantageous or convenient to substitute, delete and/or add amino acid residues to a native phosphodiesterase in order to provide convenient cloning sites in cDNA encoding the polypeptide, to aid in purification of the polypeptide, and for crystallization of the polypeptide. Such substitutions, deletions and/or additions which do not substantially alter the three dimensional structure of the native phosphodiesterase domain will be apparent to those of ordinary skill in the art.

[0210] It should be noted that the mutants conteinplated herein need not all exhibit phosphodiesterase activity. Indeed, amino acid substitutions, additions or deletions that interfere with the phosphodiesterase activity but which do not significantly alter the three-dimensional structure of the domain are specifically contemplated by the invention. Such crystalline polypeptides, or the atomic structure coordinates obtained therefrom, can be used to identify compounds that bind to the native domain. These compounds can affect the activity of the native domain.

[0211] The derivative crystals of the invention can comprise a crystalline phosphodiesterase polypeptide in covalent association with one or more heavy metal atoms. The polypeptide may correspond to a native or a mutated phosphodiesterase. Heavy metal atoms useful for providing derivative crystals include, by way of example and not limitation, gold, mercury, selenium, etc.

[0212] The co-crystals of the invention generally comprise a crystalline phosphodiesterase domain polypeptide in association with one or more compounds. The association may be covalent or non-covalent. Such coinpounds include, but are not limited to, cofactors, substrates, substrate analogues, inhibitors, allosteric effectors, etc.

III. Three Dimensional Structure Determination Using X-ray Crystallography [0213] X-ray crystallography is a method of solving the three dimensional structures of molecules. The structure of a molecule is calculated from X-ray diffraction patterns using a crystal as a diffraction grating. Three dimensional structures of protein molecules arise from crystals grown from a concentrated aqueous solution of that protein. The process of X-ray crystallography can include the following steps:

(a) synthesizing and isolating (or otherwise obtaining) a polypeptide;
(b) growing a crystal from an aqueous solution comprising the polypeptide with or without a modulator; and (c) collecting X-ray diffraction patterns from the crystals, determining unit cell dimensions and symmetry, determining electron density, fitting the amino acid sequence of the polypeptide to the electron density, and refining the structure.
Production of Polypeptides [0214] The native and mutated phosphodiesterase polypeptides described herein may be chemically synthesized in whole or part using techniques that are well-known in the art (see, e.g., Creighton (1983) Biopolynaers 22(1):49-58).

[0215] Alternatively, methods which are well known to those skilled in the art can be used to construct expression vectors containing the native or mutated phosphodiesterase polypeptide coding sequence and appropriate transcriptional/translational control signals.
These methods include in vitro recombinant DNA techniques, synthetic techniques and in vivo recoinbination/genetic recombination. See, for example, the techniques described in Maniatis, T (1989). Molecular cloning: A laboratory Manual. Cold Spring Harbor Laboratory, New York. Cold Spring Harbor Laboratory Press; and Ausubel, F.M. et al. (1994) Current Protocols in Molecular Biology. Jolui Wiley & Sons, Secaucus, N.J.

[0216] A variety of host-expression vector systems may be utilized to express the phosphodiesterase coding sequence. These include but are not limited to microorganisms such as bacteria transformed with recombinant bacteriophage DNA, plasmid DNA or cosmid DNA
expression vectors containing the phosphodiesterase domain coding sequence;
yeast transformed with recombinant yeast expression vectors containing the phosphodiesterase domain coding sequence; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing the phosphodiesterase domain coding sequence; plant cell systems infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing the phosphodiesterase domain coding sequence; or animal cell systems. The expression elements of these systems vary in their strength and specificities.

[0217] Depending on the host/vector system utilized, any of a number of suitable transcription and translation elements, including constitutive and inducible promoters, may be used in the expression vector. For example, when cloning in bacterial systems, inducible promoters such as pL of bacteriophage X, plac, ptrp, ptac (ptrp-lac hybrid promoter) and the like may be used; when cloning in insect cell systems, promoters such as the baculovirus polyhedrin promoter may be used; wlien cloning in plant cell systems, promoters derived from the genome of plant cells (e.g., heat shock promoters; the promoter for the small subunit of RUBISCO; the promoter for the chlorophyll alb binding protein) or from plant viruses (e.g., the 35S RNA promoter of CaMV; the coat protein promoter of TMV) may be used;
when cloning in mammalian cell systems, promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or from mammalian viruses (e.g., the adenovirus late promoter; the vaccinia virus 7.5K promoter) may be used; wlien generating cell lines that contain multiple copies of the phosphodiesterase domain DNA, SV4O-, BPV- and EBV-based vectors may be used with an appropriate selectable marker.

[0218] Exemplary methods describing methods of DNA manipulation, vectors, various types of cells used, methods of incorporating the vectors into the cells, expression techniques, protein purification and isolation methods, and protein concentration methods are disclosed in detail in PCT publication WO 96/18738. This publication is incorporated herein by reference in its entirety, including any drawings. Those skilled in the art will appreciate that such descriptions are applicable to the present invention and can be easily adapted to it.

Crystal Growth [0219] Crystals are grown from an aqueous solution containing the purified and concentrated polypeptide by a variety of techniques. These techniques include batch, liquid, bridge, dialysis, vapor diffusion, and hanging drop methods. McPherson (1982) John Wiley, New York;
McPherson (1990) Eur. J. Biochem. 189:1-23; Webber (1991) Adv. Proteifi Chem.
41:1-36, incorporated by reference herein in their entireties, including all figures, tables, and drawings.
[0220] The native crystals of the invention are, in general, grown by adding precipitants to the concentrated solution of the polyp eptide. The precipitants are added at a concentration just below that necessary to precipitate the protein. Water is removed by controlled evaporation to produce precipitating conditions, which are maintained until crystal growth ceases.

[0221] For crystals of the invention, exemplary crystallization conditions are described in the Examples. Those of ordinary skill in the art will recognize that the exemplary crystallization conditions can be varied. Such variations may be used alone or in combination.
In addition, other crystallization conditions may be found, e.g., by using crystallization screening plates to identify such other conditions. Those alternate conditions can then be optimized if needed to provide larger or better quality crystals.

[0222] Derivative crystals of the invention can be obtained by soaking native crystals in mother liquor containing salts of heavy metal atoms. It has been found that soaking a native crystal in a solution containing about 0.1 mM to about 5 inM thimerosal, 4-chloromeruribenzoic acid or KAu(CN)Z for about 2 hr to about 72 hr provides derivative crystals suitable for use as isomorphous replacements in determining the X-ray crystal structure.

[0223] Co-crystals of the invention can be obtained by soaking a native crystal in mother liquor containing compound that binds the phosphodiesterase, or can be obtained by co-crystallizing the phosphodiesterase polypeptide in the presence of a binding compound.
[0224] Generally, co-crystallization of phosphodiesterase and binding compound can be accomplished using conditions identified for crystallizing the corresponding phosphodiesterase without binding compound. It is advantageous if a plurality of different crystallization conditions have been identified for the phosphodiesterase, and these can be tested to determine which condition gives the best co-crystals. It may also be benficial to optimize the conditions for co-crystallization. Alternatively, new crystallization conditions can be determined for obtaining co-crystals, e.g., by screening for crystallization and then optimizing those conditions. Exemplary co-crystallization conditions are provided in the Examples.

Determining Unit Cell Dimensions and the Three Dimensional Structure of a Polypeptide or Polypeptide Complex [0225] Once the crystal is grown, it can be placed in a glass capillary tube or other mounting device and mounted onto a holding device connected to an X-ray generator and an X-ray detection device. Collection of X-ray diffraction patterns are well documented by those in the art. See, e.g., Ducruix and Geige, (1992), IRL Press, Oxford, England, and references cited therein. A beam of X-rays enters the crystal and then diffracts from the crystal. An X-ray detection device can be utilized to record the diffraction patterns emanating from the crystal.
Although the X-ray detection device on older models of these instruments is a piece of film, modern instruments digitally record X-ray diffraction scattering. X-ray sources can be of various types, but advantageously, a high intensity source is used, e.g., a synchrotron beam source.

[0226] Methods for obtaining the three dimensional structure of the crystalline form of a peptide molecule or molecule complex are well known in the art. See, e.g., Ducruix and Geige, (1992), IIZL Press, Oxford, England, and references cited therein. The following are steps in the process of determining the three dimensional structure of a molecule or complex from X-ray diffraction data.

[0227] After the X-ray diffraction patterns are collected from the crystal, the unit cell dimensions and orientation in the crystal can be determined. They can be determined from the spacing between the diffraction emissions as well as the patterns made from these emissions.
The unit cell dimensions are characterized in three dimensions in units of Angstroms (one A=
10-10 meters) and by angles at each vertices. The symmetry of the unit cell in the crystals is also characterized at this stage. The symmetry of the unit cell in the crystal simplifies the complexity of the collected data by identifying repeating patterns.
Application of the symmetry and dimensions of the unit cell is described below.

[0228] Each diffraction pattern emission is characterized as a vector and the data collected at this stage of the method determines the amplitude of each vector. The phases of the vectors can be determined using multiple techniques. In one method, heavy atoms can be soaked into a crystal, a method called isomorphous replacement, and the phases of the vectors can be detennined by using these heavy atoms as reference points in the X-ray analysis. (Otwinowski, (1991), Daresbury, United Kingdom, 80-86). The isomorphous replaceinent method usually utilizes more than one heavy atom derivative.

[0229] In another method, the amplitudes and phases of vectors from a crystalline polypeptide with an already determined structure can be applied to the amplitudes of the vectors from a crystalline polypeptide of unknown structure and consequently determine the phases of these vectors. This second method is known as molecular replacement and the protein structure which is used as a reference must have a closely related structure to the protein of interest. (Naraza (1994) Proteins 11:281-296). Thus, the vector information from a phosphodiesterase of known structure, such as those reported herein, are useful for the molecular replacement analysis of another phosphodiesterase with unknown structure.

[0230] Once the phases of the vectors describing the unit cell of a crystal are determined, the vector amplitudes and phases, unit cell dimensions, and unit cell symmetry can be used as terms in a Fourier transform function. The Fourier transform function calculates the electron density in the unit cell from these measurements. The electron density that describes one of the molecules or one of the molecule complexes in the unit cell can be referred to as an electron density map. The amino acid structures of the sequence or the molecular structures of compounds complexed with the crystalline polypeptide may then be fitted to the electron density using a variety of computer programs. This step of the process is sometimes referred to as model building and can be accomplished by using computer programs such as Turbo/FRODO or "0". (Jones (1985) Methads in Enzynaology 115:157-171).

[0231] A theoretical electron density map can then be calculated from the amino acid structures fit to the experimentally determined electron density. The theoretical and experimental electron density maps can be compared to one another and the agreement between these two maps can be described by a parameter called an R-factor. A
low value for an R-factor describes a high degree of overlapping electron density between a theoretical and experimental electron density map.

[0232] The R-factor is then minimized by using computer programs that refine the theoretical electron density map. A computer program such as X-PLOR can be used for model refinement by those skilled in the art. (Brunger (1992) Nature 355:472-475.) Refinement may be achieved in an iterative process. A first step can entail altering the conformation of atoms defined in an electron density map. The conformations of the atoms can be altered by simulating a rise in temperature, which will increase the vibrational frequency of the bonds and modify positions of atoms in the structure. At a particular point in the atomic perturbation process, a force field, which typically defines interactions between atoms in terms of allowed bond angles and bond lengths, Van der Waals interactions, hydrogen bonds, ionic interactions, and hydrophobic interactions, can be applied to the system of atoms. Favorable interactions may be described in terms of free energy and the atoms can be moved over many iterations until a free energy minimum is achieved. The refinement process can be iterated until the R-factor reaches a minimum value.

[0233] The three dimensional structure of the molecule or molecule complex is described by atoms that fit the theoretical electron density characterized by a minimum R-value. A file can then be created for the three dimensional structure that defines each atom by coordinates in three dimensions. An example of such a structural coordinate file is shown in Table 1.

IV. Structures of PDE4B

[0234] High-resolution three-dimensional structures and atomic structure coordinates of crystalline PDE4B phosphodiesterase domain and PDE4B phosphodiesterase domain co-complexed with exemplary binding coinpounds are described. The methods used to obtain the structure coordinates are provided in the examples. The atomic structure coordinates of crystalline PDE4B phosphodiesterase domain are listed in Table 1. Co-crystal coordinates can be used in the same way, e.g., in the various aspects described herein, as coordinates for the protein by itself, but can be advantageous because such co-crystals demonstrate or confirm the binding mode of binding compound, and can also include shifts of protein atoms in response to the presence of the binding compound.

[0235] Those having skill in the art will recognize that atomic structure coordinates as determined by X-ray crystallography are not without error. Thus, it is to be understood that generally any set of structure coordinates obtained for crystals of PDE, whether native crystals, phosphodiesterase domain crystals, derivative crystals or co-crystals, that have a root mean square deviation ("r.m.s.d.") of less than or equal to about 1.5 A when superimposed, using backbone atoms (N, Ca,, C and 0), on the structure coordinates listed in a coordinate table herein are considered to be identical witli the structure coordinates listed in that table when at least about 50% to 100% of the backbone atoms of the crystallized protein are included in the superposition.

V. Uses of the Crystals and Atomic Structure Coordinates [0236] The crystals of the invention, and particularly the atomic structure coordinates obtained therefrom, have a wide variety of uses. For example, the crystals described herein can be used as a starting point in any of the methods of use for phosphodiesterases known in the art or later developed. Such methods of use include, for example, identifying molecules that bind to the native or mutated catalytic domain of phosphodiesterases. The crystals and structure coordinates are particularly useful for identifying ligands that modulate phosphodiesterase activity as an approach towards developing new therapeutic agents. In particular, the crystals and structural information are useful in methods for ligand development utilizing molecular scaffolds.

[0237] The structure coordinates described herein can be used as phasing models for determining the crystal structures of additional phosphodiesterases, as well as the structures of co-crystals of such phosphodiesterases with ligands such as inhibitors, agonists, antagonists, and other molecules. The structure coordinates, as well as models of the three-dimensional structures obtained therefrom, can also be used to aid the elucidation of solution-based structures of native or mutated phosphodiesterases, such as those obtained via NMR.

VI. Electronic Representations of Phosphodiesterase Structures [0238] Structural information of phosphodiesterases or portions of phosphodiesterases (e.g., phosphodiesterase active sites) can be represented in many different ways.
Particularly useful are electronic representations, as such representations allow rapid and convenient data manipulations and structural modifications. Electronic representations can be embedded in many different storage or memory media, frequently computer readable media.
Examples include without limitations, computer random access memory (RAM), floppy disk, magnetic hard drive, magnetic tape (analog or digital), compact disk (CD), optical disk, CD-ROM, memory card, digital video disk (DVD), and others. The storage medium can be separate or part of a computer system. Such a computer system may be a dedicated, special purpose, or embedded system, such as a computer system that forms part of an X-ray crystallography system, or may be a general purpose computer (which may have data connection with other equipment such as a sensor device in an X-ray crystallographic system. In many cases, the information provided by such electronic representations can also be represented physically or visually in two or three dimensions, e.g., on paper, as a visual display (e.g., on a computer monitor as a two dimensional or pseudo-three dimensional image) or as a three dimensional physical model. Such physical representations can also be used, alone or in connection with electronic representations. Exemplary useful representations include, but are not limited to, the following:

Atomic Coordinate Representation [0239] One type of representation is a list or table of atomic coordinates representing positions of particular atoms in a molecular structure, portions of a structure, or complex (e.g., a co-crystal). Such a representation may also include additional information, for example, information about occupancy of particular coordinates. One such atomic coordinate representation contains the coordinate information of Table 1 in electronic form.

Energy Surface or Surface of Interaction Representation [0240] Another representation is an energy surface representation, e.g., of an active site or other binding site, representing an energy surface for electronic and steric interactions. Such a representation may also include other features. An example is the inclusion of representation of a particular amino acid residue(s) or group(s) on a particular amino acid residue(s), e.g., a residue or group that can participate in H-bonding or ionic interaction. Such energy surface representations can be readily generated from atomic coordinate representations using any of a variety of available computer programs.

Structural Representation [0241] Still another representation is a structural representation, i.e., a physical representation or an electronic representation of such a physical representation. Such a structural representation includes representations of relative positions of particular features of a molecule or complex, often with linkage between structural features. For example, a structure can be represented in which all atoms are linked; atoms other than hydrogen are linked; backbone atoms, with or without representation of sidechain atoms that could participate in significant electronic interaction, are linked; among others. However, not all features need to be linked.
For example, for structural representations of portions of a molecule or complex, structural features significant for that feature may be represented (e.g., atoms of amino acid residues that can have significant binding interation with a ligand at a binding site. Those amino acid residues may not be linked with each other.

[0242] A structural representation can also be a schematic representation. For example, a schematic representation can represent secondary and/or tertiary structure in a schematic manner. Within such a schematic representation of a polypeptide, a particular amino acid residue(s) or group(s) on a residue(s) can be included, e.g., conserved residues in a binding site, and/or residue(s) or group(s) that may interact with binding compounds.
Electronic structural representations can be generated, for example, from atomic coordinate information using computer programs designed for that function and/or by constructing an electronic representation with manual input based on interpretation of another form of structural information. Physical representations can be created, for example, by printing an image of a computer-generated image or by constructing a 3D model. An example of such a printed representation is the ribbon diagram presented in Figure 2.

VII. Structure Determination for Phosphodiesterases with Unknown Structure Using Structural Coordinates [0243] Structural coordinates, such as those set forth in Table 1, can be used to determine the three dimensional structures of phosphodiesterases with unknown structure. The methods described below can apply structural coordinates of a polypeptide with known structure to another data set, such as an amino acid sequence, X-ray crystallographic diffraction data, or nuclear magnetic resonance (NMR) data. Preferred embodiments of the invention relate to determining the three dimensional structures of modified phosphodiesterases, other native phosphodiesterases, and related polypeptides.

Structures Using Amino Acid Homology [0244] Homology modeling is a metllod of applying structural coordinates of a polypeptide of known structure to the amino acid sequence of a polypeptide of unknown structure. This method is accomplished using a computer representation of the three dimensional structure of a polypeptide or polypeptide complex, the computer representation of amino acid sequences of the polypeptides with known and unknown structures, and standard computer representations of the structures of amino acids. Homology modeling generally involves (a) aligning the amino acid sequences of the polypeptides with and without known structure; (b) transferring the coordinates of the conserved amino acids in the known stracture to the corresponding ainino acids of the polypeptide of unknown structure; refining the subsequent three dimensional structure; and (d) constructing structures of the rest of the polypeptide. One skilled in the art recognizes that conserved amino acids between two proteins can be determined from the sequence alignment step in step (a).

[0245] The above method is well known to those skilled in the art. (Greer (1985) Science 228:1055; Blundell et al. A(1988) Eur. J. Biochena. 172:513. An exemplary computer program that can be utilized for homology modeling by those skilled in the art is the Homology module in the Insight II modeling package distributed by Accelerys Inc.

[0246] Alignment of the amino acid sequence is accomplished by first placing the computer representation of the amino acid sequence of a polypeptide with known structure above the amino acid sequence of the polypeptide of unknown structure. Amino acids in the sequences are then compared and groups of amino acids that are homologous (e.g., amino acid side chains that are similar in chemical nature - aliphatic, aromatic, polar, or charged) are grouped together. This method will detect conserved regions of the polypeptides and account for amino acid insertions or deletions. Such alignment and/or can also be performed fully computationally using sequence alignment and analysis software.

[0247] Once the amino acid sequences of the polypeptides with known and unknown structures are aligned, the structures of the conserved amino acids in the computer representation of the polypeptide with known structure are transferred to the corresponding amino acids of the polypeptide whose structure is unknown. For example, a tyrosine in the amino acid sequence of known structure may be replaced by a phenylalanine, the corresponding homologous amino acid in the amino acid sequence of unknown structure.
[0248] The structures of amino acids located in non-conserved regions are to be assigned manually by either using standard peptide geometries or molecular simulation techniques, such as molecular dynamics. The final step in the process is accomplished by refining the entire structure using molecular dynamics and/or energy minimization. The homology modeling method is well known to those skilled in the art and has been practiced using different protein molecules. For example, the three dimensional structure of the polypeptide corresponding to the catalytic domain of a serine/threonine protein kinase, myosin light chain protein kinase, was homology modeled from the cAMP-dependent protein kinase catalytic subunit.
(Knighton et al. (1992) Science 258:130-135.) Structures Using Molecular Replacement [0249] Molecular replacement is a method of applying the X-ray diffraction data of a polypeptide of known structure to the X-ray diffraction data of a polypeptide of unknown sequence. This method can be utilized to define the phases describing the X-ray diffraction data of a polypeptide of unknown structure when only the amplitudes are known.
X-PLOR is a commonly utilized computer software package used for molecular replacement.
Brunger (1992) Nature 355:472-475. AMORE is another program used for molecular replacement.

Navaza (1994) Acta Crystallogr. A50:157-163. Preferably, the resulting structure does not exhibit a root-mean-square deviation of more than 3A.

[0250] A goal of molecular replacement is to align the positions of atoms in the unit cell by matching electron diffraction data from two crystals. A program such as X-PLOR
can involve four steps. A first step can be to determine the number of molecules in the unit cell and define the angles between them. A second step can involve rotating the diffraction data to define the orientation of the molecules in the unit cell. A third step can be to translate the electron density in three dimensions to correctly position the molecules in the unit cell. Once the amplitudes and phases of the X-ray diffraction data is determined, an R-factor can be calculated by comparing electron diffraction maps calculated experimentally from the reference data set and calculated from the new data set. An R-factor between 30-50% indicates that the orientations of the atoms in the unit cell are reasonably determined by this method. A
fourth step in the process can be to decrease the R-factor to roughly 20% by refining the new electron density map using iterative refinement techniques described herein and known to those or ordinary skill in the art.

Structures Using NMR Data [0251] Structural coordinates of a polypeptide or polypeptide complex derived from X-ray crystallographic techniques can be applied towards the elucidation of three dimensional structures of polypeptides from nuclear magnetic resonance (NMR) data. This method is used by those skilled in the art. (Wuthrich, (1986), John Wiley and Sons, New York:176-199;
Pflugrath et al. (1986) J. Mol. Biol. 189:383-386,= Kline et al. (1986) J.
Mol. Biol. 189:377-382.) While the secondary structure of a polypeptide is often readily determined by utilizing two-dimensional NMR data, the spatial connections between individual pieces of secondary structure are not as readily determinable. The coordinates defining a three-dimensional structure of a polypeptide derived from X-ray crystallographic techniques can guide the NMR
spectroscopist to an understanding of these spatial interactions between secondary structural elements in a polypeptide of related structure.

[0252] The knowledge of spatial interactions between secondary structural elements can greatly simplify Nuclear Overhauser Effect (NOE) data from two-dimensional NMR
experiments. Additionally, applying the crystallographic coordinates after the determination of secondary structure by NMR techniques only simplifies the assignment of NOEs relating to particular amino acids in the polypeptide sequence and does not greatly bias the NMR
analysis of polypeptide structure. Conversely, using the crystallographic coordinates to simplify NOE data while determining secondary structure of the polypeptide would bias the NMR analysis of protein structure.

VIII. Structure-Based Design of Modulators of Phosphodiesterase Function Utilizing Structural Coordinates [0253] Structure-based modulator design and identification methods are powerful techniques that can involve searches of computer databases containing a wide variety of potential modulators and chemical functional groups. The computerized design and identification of modulators is useful as the computer databases contain more compounds than the chemical libraries, often by an order of magnitude. For reviews of structure-based drug design and identification (see Kuntz et al. (1994), Acc. Chem. Res. 27:117; Guida (1994) Current Opinion in Struc. Biol. 4: 777; Colman (1994) Current Opinion in. Struc. Biol. 4:
868).

[0254] The three dimensional structure of a polypeptide defined by structural coordinates can be utilized by these design methods, for example, the structural coordinates of Table 1. In addition, the three dimensional structures of phosphodiesterases determined by the homology, molecular replacement, and NMR techniques described herein can also be applied to modulator design and identification methods.

[0255] For identifying modulators, structural information for a native phosphodiesterase, in particular, structural information for the active site of the phosphodiesterase, can be used.
However, it may be advantageous to utilize structural information from one or more co-crystals of the phosphodiesterase with one or more binding compounds. It can also be advantageous if the binding compound has a structural core in common with test compounds.

Design by Searching Molecular Data Bases [0256] One method of rational design searches for modulators by docking the computer representations of compounds from a database of molecules. Publicly available databases include, for example:
a) ACD from Molecular Designs Limited b) NCI from National Cancer Institute c) CCDC from Cambridge Crystallographic Data Center d) CAST from Chemical Abstract Service e) Derwent from Derwent Information Limited f) Maybridge from Maybridge Chemical Company LTD
g) Aldrich from Aldrich Chemical Company h) Directory of Natural Products from Chapman & Hall [0257] One such data base (ACD distributed by Molecular Designs Limited Information Systems) contains compounds that are synthetically derived or are natural products. Methods available to those skilled in the art can convert a data set represented in two dimensions to one represented in three diinensions. These methods are enabled by such computer programs as CONCORD from Tripos Associates or DE-Converter from Molecular Simulations Limited.
[0258] Multiple methods of structure-based modulator design are known to those in the art.
(Kuntz et al., (1982), J. Mol. Biol. 162: 269; Kuntz et aZ., (1994), Acc.
Chern. Res. 27: 117;
Meng et al., (1992), J Compt. Chem. 13: 505; Bohm, (1994), J. Comp. Aided Molec. Design 8: 623.) [0259] A computer prograin widely utilized by those skilled in the art of rational modulator design is DOCK from the University of California in San Francisco. The general methods utilized by this computer program and programs like it are described in three applications below. More detailed information regarding some of these techniques can be found in the Accelerys User Guide, 1995. A typical computer program used for this purpose can perform a processes comprising the following steps or functions:
(a) remove the existing compound from the protein;
(b) dock the structure of another compound into the active-site using the computer program (such as DOCK) or by interactively moving the compound into the active-site;
(c) characterize the space between the compound and the active-site atoms;
(d) search libraries for molecular fraginents which (i) can fit into the empty space between the compound and the active-site, and (ii) can be linked to the compound;
and (e) link the fragments found above to the compound and evaluate the new modified compound.

[0260] Part (c) refers to characterizing the geometry and the complementary interactions formed between the atoms of the active site and the compounds. A favorable geometric fit is attained when a significant surface area is shared between the compound and active-site atoms without forming unfavorable steric interactions. One skilled in the art would note that the method can be performed by skipping parts (d) and (e) and screening a database of many compounds.

[0261] Structure-based design and identification of modulators of phosphodiesterase function can be used in conjunction with assay screening. As large computer databases of compounds (around 10,000 compounds) can be searched in a matter of hours or even less, the computer-based method can narrow the compounds tested as potential modulators of phosphodiesterase function in biochemical or cellular assays.

[0262] The above descriptions of structure-based modulator design are not all encompassing and other methods are reported in the literature and can be used, e.g.:
(1) CAVEAT: Bartlett et al.,(1989), in Chemical and Biological Problems in Molecular Recognition, Roberts, S.M.; Ley, S.V.; Campbell, M.M. eds.; Royal Society of Chemistry: Cambridge, pp.182-196.
(2) FLOG: Miller et al., (1994), J Comp. Aided Molec. Design 8:153.
(3) PRO Modulator: Clark et al., (1995), J. Comp. Aided Molec. Design 9:13.
(4) MCSS: Miranker and Karplus, (1991), Proteins: Structure, Function, and Genetics 11:29.
(5) AUTODOCK: Goodsell and Olson, (1990), Proteins: Structure, Function, and Genetics 8:195.
(6) GRID: Goodford, (1985), J. Med. Chem. 28:849.

Design by Modifying Compounds in Complex with PDE4B

[0263] Another way of identifying compounds as potential modulators is to modify an existing modulator in the polypeptide active site. For example, the computer representation of modulators can be modified within the coinputer representation of a PDE4B
active site.
Detailed instructions for this technique can be found, for example, in the Accelerys User Manual, 1995 in LUDI. The computer representation of the modulator is typically modified by the deletion of a chemical group or groups or by the addition of a chemical group or groups.
[0264] Upon each modification to the compound, the atoms of the modified compound and active site can be shifted in conformation and the distance between the modulator and the active-site atoms may be scored along with any complementary interactions formed between the two molecules. Scoring can be complete when a favorable geometric fit and favorable complementary interactions are attained. Compounds that have favorable scores are potential modulators.

Design by Modifying the Structure of Compounds that Bind PDE4B

[0265] A third method of structure-based modulator design is to screen compounds designed by a modulator building or modulator searching computer program. Examples of these types of programs can be found in the Molecular Simulations Package, Catalyst.
Descriptions for using this program are documented in the Molecular Siinulations User Guide (1995).
Other computer programs used in this application are ISIS/HOST, ISIS/BASE, ISIS/DRAW) from Molecular Designs Limited and UNITY from Tripos Associates.

[0266] These programs can be operated on the structure of a compound that has been removed from the active site of the three dimensional structure of a compound-phosphodiesterase complex. Operating the prograin on such a compound is preferable since it is in a biologically active conformation.

[0267] A modulator construction computer program is a computer program that may be used to replace computer representations of chemical groups in a compound complexed with a phosphodiesterase or other biomolecule with groups from a computer database. A
modulator searching computer program is a computer program that may be used to search computer representations of compounds from a computer data base that have similar three dimensional structures and similar chemical groups as compound bound to a particular biomolecule.

[0268] A typical program can operate by using the following general steps:
(a) map the compounds by chemical features such as by hydrogen bond donors or acceptors, hydrophobic/lipophilic sites, positively ionizable sites, or negatively ionizable sites;
(b) add geometric constraints to the mapped features; and (c) search databases with the model generated in (b).

[0269] Those skilled in the art also recognize that not all of the possible chemical features of the compound need be present in the model of (b). One can use any subset of the model to generate different models for data base searches.

Modulator Design Using Molecular Scaffolds [0270] The present invention can also advantageously utilize methods for designing compounds, designated as molecular scaffolds, that can act broadly across families of molecules and/or for using a molecular scaffold to design ligands that target individual or multiple members of those families. Such design using molecular scaffolds is described in Hirth and Milburn, U.S. Patent Application 10/377,268, which is incorporated herein by reference in its entirety. Such design and development using molecular scaffolds is described, in part, below.

[02711 In preferred embodiments, the molecules can be proteins and a set of chemical compounds can be assembled that have properties such that they are 1) chemically designed to act on certain protein families and/or 2) behave more like molecular scaffolds, meaning that they have chemical substructures that make them specific for binding to one or more proteins in a family of interest. Alternatively, molecular scaffolds can be designed that are preferentially active on an individual target molecule.

[0272] Useful chemical properties of molecular scaffolds can include one or more of the following characteristics, but are not limited thereto: an average molecular weight below about 350 daltons, or between from about 150 to about 350 daltons, or from about 150 to about 300 daltons; having a clogP below 3; a number of rotatable bonds of less than 4; a number of hydrogen bond donors and acceptors below 5 or below 4; a polar surface area of less than 50 A2; binding at protein binding sites in an orientation so that chemical substituents from a combinatorial library that are attached to the scaffold can be projected into pockets in the protein binding site; and possessing chemically tractable structures at its substituent attachment points that can be modified, thereby enabling rapid library construction.

[0273] By "clog P" is meant the calculated log P of a compound, "P" referring to the partition coefficient between octanol and water.

[0274] The term "Molecular Polar Surface Area (PSA)" refers to the sum of surface contributions of polar atoms (usually oxygens, nitrogens and attached hydrogens) in a molecule. The polar surface area has been shown to correlate well with drug transport properties, such as intestinal absorption, or blood-brain barrier penetration.

[0275] Additional useful chemical properties of distinct compounds for inclusion in a combinatorial library include the ability to attach chemical moieties to the compound that will not interfere with binding of the compound to at least one protein of interest, and that will impart desirable properties to the library members, for example, causing the library members to be actively transported to cells and/or organs of interest, or the ability to attach to a device such as a chromatography column (e.g., a streptavidin column through a molecule such as biotin) for uses such as tissue and proteomics profiling purposes.

[0276] A person of ordinary skill in the art will realize other properties that can be desirable for the scaffold or library members to have depending on the particular requirements of the use, and that compounds with these properties can also be sought and identified in like manner.
Methods of selecting compounds for assay are known to those of ordinary skill in the art, for example, methods and compounds described in U.S. Patent No. 6,288,234, 6,090,912, 5,840,485, each of which is hereby incorporated by reference in its entirety, including all charts and drawings.

[0277] In various embodiments, the present invention provides methods of designing ligands that bind to a plurality of members of a molecular family, where the ligands contain a common molecular scaffold. Thus, a compound set can be assayed for binding to a plurality of members of a molecular family, e.g., a protein family. One or more compounds that bind to a plurality of family members can be identified as molecular scaffolds. When the orientation of the scaffold at the binding site of the target molecules has been determined and chemically tractable structures have been identified, a set of ligands can be synthesized starting with one or a few molecular scaffolds to arrive at a plurality of ligands, wherein each ligand binds to a separate target molecule of the molecular family with altered or changed binding affinity or binding specificity relative to the scaffold. Thus, a plurality of drug lead molecules can be designed to preferentially target individual members of a molecular family based on the same molecular scaffold, and act on them in a specific manner.

IX. Binding Assays [0278] The methods of the present invention can involve assays that are able to detect the binding of compounds to a target molecule. Such binding is at a statistically significant level, preferably with a confidence level of at least 90%, more preferably at least 95, 97, 98, 99% or greater confidence level that the assay signal represents binding to the target molecule, i.e., is distinguished from background. Preferably controls are used to distinguish target binding from non-specific binding. The assays of the present invention can also include assaying compounds for low affinity binding to the target molecule. A large variety of assays indicative of binding are known for different target types and can be used for this invention. Compounds that act broadly across protein fanlilies are not likely to have a high affinity against individual targets, due to the broad nature of their binding. Thus, assays described herein allow for the identification of compounds that bind with low affinity, very low affinity, and extremely low affinity. Therefore, potency (or binding affinity) is not the primary, nor even the most iinportant, indicia of identification of a potentially useful binding compound. Rather, even those compounds that bind with low affinity, very low affinity, or extremely low affinity can be considered as molecular scaffolds that can continue to the next phase of the ligand design process.

[0279] By binding with "low affmity" is meant binding to the target molecule with a dissociation constant (kd) of greater than 1 M under standard conditions. By binding with "very low affinity" is meant binding with a kd of above about 100 M under standard conditions. By binding with "extremely low affinity" is meant binding at a kd of above about 1 mM under standard conditions. By "moderate affinity" is meant binding with a kd of from about 200 nM to about 1 M under standard conditions. By "moderately high affmity" is meant binding at a kd of from about 1 nM to about 200 nM. By binding at "high affinity" is meant binding at a kd of below about 1 nM under standard conditions. For example, low affinity binding can occur because of a poorer fit into the binding site of the target molecule or because of a smaller number of non-covalent bonds, or weaker covalent bonds present to cause binding of the scaffold or ligand to the binding site of the target molecule relative to instances where higher affinity binding occurs. The standard conditions for binding are at pH 7.2 at 37 C for one hour. For example, 100 l/well can be used in HEPES 50 mM buffer at pH 7.2, NaCI 15 mM, ATP 2 M, and bovine serum albumin 1 ug/well, 37 C for one hour.

[0280] Binding compounds can also be characterized by their effect on the activity of the target molecule. Thus, a "low activity" compound has an inhibitory concentration (IC50) or excitation concentration (EC50) of greater than 1 M under standard conditions. By "very low activity" is meant an IC50 or EC50 of above 100 M under standard conditions.
By "extremely low activity" is meant an IC50 or EC50 of above 1 mM under standard conditions. By "moderate activity" is meant an IC50 or EC50 of 200 nM to 1 M under standard conditions.

By "moderately high activity" is meant an IC50 or EC50 of 1 nM to 200 nM. By "high activity" is meant an IC50 or EC50 of below 1 nM under standard conditions.
The IC50 (or EC50) is defined as the concentration of compound at which 50% of the activity of the target molecule (e.g., enzyme or other protein) activity being measured is lost (or gained) relative to activity when no compound is present. Activity can be measured using methods known to those of ordinary skill in the art, e.g., by measuring any detectable product or signal produced by occurrence of an enzymatic reaction, or other activity by a protein being measured.

[0281] By "background signal" in reference to a binding assay is meant the signal that is recorded under standard conditions for the particular assay in the absence of a test compound, molecular scaffold, or ligand that binds to the target molecule. Persons of ordinary skill in the art will realize that accepted methods exist and are widely available for determining background signal.

[0282] By "standard deviation" is meant the square root of the variance. The variance is a measure of how spread out a distribution is. It is computed as the average squared deviation of each number from its mean. For example, for the numbers 1, 2, and 3, the mean is 2 and the variance is:
aZ = (1-2)2 + (2-2) 2 + (3-2) 2 = 0.667 [0283] To design or discover scaffolds that act broadly across protein families, proteins of interest can be assayed against a compound collection or set. The assays can preferably be enzymatic or binding assays. In some embodiments it may be desirable to enhance the solubility of the compounds being screened and then analyze all compounds that show activity in the assay, including those that bind with low affinity or produce a signal with greater than about three times the standard deviation of the background signal. The assays can be any suitable assay such as, for example, binding assays that measure the binding affinity between two binding partners. Various types of screening assays that can be useful in the practice of the present invention are known in the art, such as those described in U.S.
Patent Nos.
5,763,198, 5,747,276, 5,877,007, 6,243,980, 6,294,330, and 6,294,330, each of which is hereby incorporated by reference in its entirety, including all charts and drawings.

[0284] In various embodiments of the assays at least one compound, at least about 5%, at least about 10%, at least about 15%, at least about 20%, or at least about 25%
of the compounds can bind with low affinity. In general, up to about 20% of the compounds can show activity in the screening assay and these compounds can then be analyzed directly with high-throughput co-crystallography, computational analysis to group the compounds into classes with common structural properties (e.g., structural core and/or shape and polarity characteristics), and the identification of common chemical structures between compounds that show activity.

[0285] The person of ordinary skill in the art will realize that decisions can be based on criteria that are appropriate for the needs of the particular situation, and that the decisions can be made by computer software programs. Classes can be created containing almost any number of scaffolds, and the criteria selected can be based on increasingly exacting criteria until an arbitrary number of scaffolds is arrived at for each class that is deemed to be advantageous.

Surface Plasmon Resonance [0286] Binding paraineters can be measured using surface plasmon resonance, for example, with a BIAcore chip (Biacore, Japan) coated with immobilized binding components. Surface plasmon resonance is used to characterize the microscopic association and dissociation constants of reaction between an sFv or otlier ligand directed against target molecules. Such methods are generally described in the following references which are incorporated herein by reference. Vely F. et al., (2000) BIAcore analysis to test phosphopeptide-SH2 domain interactions, Methods in Molecular Biology. 121:313-21; Liparoto et al., (1999) Biosensor analysis of the interleukin-2 receptor complex, Journal of Molecular Recognition. 12:316-21;
Lipschultz et al., (2000) Experimental design for analysis of complex kinetics using surface plasmon resonance, Methods. 20(3):310-8; Malmqvist., (1999) BIACORE: an affinity biosensor system for characterization of biomolecular interactions, Biochemical Society Transactions 27:335-40; Alfthan, (1998) Surface plasmon resonance biosensors as a tool in antibody engineering, Biosensors & BioelectYonics. 13:653-63; Fivash et al., (1998) BlAcore for macromolecular interaction, Current Opinion in Biotechnology. 9:97-101;
Price et al.;
(1998) Summary report on the ISOBM TD-4 Workshop: analysis of 56 monoclonal antibodies against the MUC1 mucin. Tumour Biology 19 Suppl 1:1-20; Malmqvist et al, (1997) Biomolecular interaction analysis: affinity biosensor technologies for functional analysis of proteins, Current Opinion in C/zemical Biology. 1:378-83; O'Shannessy et al., (1996) Interpretation of deviations from pseudo-first-order kinetic behavior in the characterization of ligand binding by biosensor technology, Analytical Biochemistry. 236:275-83;
Malmborg et al., (1995) BIAcore as a tool in antibody engineering, Journal of Imrnunological Methods.
183:7-13; Van Regenmortel, (1994) Use of biosensors to characterize recombinant proteins, Developments in Biological Standardization. 83:143-5 1; and O'Shannessy, (1994) Determination of kinetic rate and equilibrium binding constants for macromolecular interactions: a critique of the surface plasmon resonance literature, Cur=rent Opinions in Biotechnology. 5:65-71.

[0287] BlAcore uses the optical properties of surface plasmon resonance (SPR) to detect alterations in protein concentration bound to a dextran matrix lying on the surface of a gold/glass sensor chip interface, a dextran biosensor matrix. In brief, proteins are covalently bound to the dextran matrix at a known concentration and a ligand for the protein is injected through the dextran matrix. Near infrared light, directed onto the opposite side of the sensor chip surface is reflected and also induces an evanescent wave in the gold filin, wliich in turn, causes an intensity dip in the reflected liglit at a particular angle known as the resonance angle.
If the refractive index of the sensor chip surface is altered (e.g., by ligand binding to the bound protein) a shift occurs in the resonance angle. This angle shift can be measured and is expressed as resonance units (RUs) such that 1000 RUs is equivalent to a change in surface protein concentration of 1 ng/mm2. These changes are displayed with respect to time along the y-axis of a sensorgram, which depicts the association and dissociation of any biological reaction.

High Throughput Screening (HTS) Assays [0288] HTS typically uses automated assays to search through large numbers of compounds for a desired activity. Typically HTS assays are used to find new drugs by screening for chemicals that act on a particular enzyme or molecule. For example, if a chemical inactivates an enzyme it might prove to be effective in preventing a process in a cell which causes a disease. High throughput methods enable researchers to assay thousands of different chemicals against each target molecule very quickly using robotic handling systems and automated analysis of results.

[0289] As used herein, "high tliroughput screening" or "HTS" refers to the rapid in vitro screening of large numbers of compounds (libraries); generally tens to hundreds of thousands of compounds, using robotic screening assays. Ultra high-throughput Screening (uHTS) generally refers to the high-throughput screening accelerated to greater than 100,000 tests per day.

[0290] To achieve high-throughput screening, it is advantageous to house samples on a multicontainer carrier or platfonn. A multicontainer carrier facilitates measuring reactions of a plurality of candidate compounds simultaneously. Multi-well microplates may be used as the carrier. Such multi-well microplates, and methods for their use in numerous assays, are both known in the art and commercially available.

[0291] Screening assays may include controls for purposes of calibration and confirmation of proper manipulation of the components of the assay. Blank wells that contain all of the reactants but no member of the chemical library are usually included. As another example, a known inhibitor (or activator) of an enzyme for which modulators are sought, can be incubated with one sample of the assay, and the resulting decrease (or increase) in the enzyme activity used as a comparator or control. It will be appreciated that modulators can also be combined with the enzyme activators or inhibitors to find modulators which inhibit the enzyme activation or repression that is otherwise caused by the presence of the known the enzyme modulator.
Similarly, when ligands to a sphingolipid target are sought, known ligands of the target can be present in control/calibration assay wells.

Measuring Enzymatic and Binding Reactions During Screening Assays [0292] Techniques for measuring the progression of enzymatic and binding reactions, e.g., in multicontainer carriers, are known in the art and include, but are not limited to, the following.
[0293] Spectrophotometric and spectrofluorometric assays are well known in the art.
Examples of such assays include the use of colorimetric assays for the detection of peroxides, as described in Gordon, A. J. and Ford, R. A., (1972) The Chemist's Companion:
A Handbook Of Practical Data, Techniques, And References, John Wiley and Sons, N.Y., Page 437.

[0294] Fluorescence spectrometry may be used to monitor the generation of reaction products. Fluorescence methodology is generally more sensitive than the absorption methodology. The use of fluorescent probes is well known to those skilled in the art. For reviews, see Bashford et al., (1987) SpectrophotomeD:y and Spectrofluorometry:
A Practical Approach, pp. 91-114, IRL Press Ltd.; and Bell, (1981) Spectroscopy In Biochemistry, Vol. I, pp. 155-194, CRC Press.

[0295] In spectrofluorometric methods, enzymes are exposed to substrates that change their intrinsic fluorescence when processed by the target enzylne. Typically, the substrate is nonfluorescent and is converted to a fluorophore through one or more reactions. As a non-limiting example, SMase activity can be detected using the Amplex Red reagent (Molecular Probes, Eugene, OR). In order to measure sphingomyelinase activity using Amplex Red, the following reactions occur. First, SMase hydrolyzes sphingomyelin to yield ceramide and phosphorylcholine. Second, alkaline phosphatase lzydrolyzes phosphorylcholine to yield choline. Third, choline is oxidized by choline oxidase to betaine. Finally, H202, in the presence of horseradish peroxidase, reacts with Amplex Red to produce the fluorescent product, Resorufin, and the signal therefrom is detected using spectrofluorometry.

[0296] Fluorescence polarization (FP) is based on a decrease in the speed of molecular rotation of a fluorophore that occurs upon binding to a larger molecule, such as a receptor protein, allowing for polarized fluorescent emission by the bound ligand. FP
is empirically determined by measuring the vertical and horizontal components of fluorophore emission following excitation with plane polarized light. Polarized emission is increased when the molecular rotation of a fluorophore is reduced. A fluorophore produces a larger polarized signal when it is bound to a larger molecule (i.e. a receptor), slowing molecular rotation of the fluorophore. The magnitude of the polarized signal relates quantitatively to the extent of fluorescent ligand binding. Accordingly, polarization of the "bound" signal depends on maintenance of high affinity binding.

[0297] FP is a homogeneous technology and reactions are very rapid, taking seconds to minutes to reach equilibrium. The reagents are stable, and large batches may be prepared, resulting in high reproducibility. Because of these properties, FP has proven to be highly automatable, often performed with a single incubation with a single, premixed, tracer-receptor reagent. For a review, see Owickiet al., (1997), Application of Fluorescence Polarization Assays in High-Throughput Screening, Genetic Engineering News, 17:27.

[0298] FP is particularly desirable since its readout is independent of the emission intensity (Checovich, W. J., et al., (1995) Nature 375:254-256; Dandliker, W. B., et al., (1981) Methods in Enzymology 74:3-28) and is thus insensitive to the presence of colored compounds that quench fluorescence emission. FP and FRET (see below) are well-suited for identifying compounds that block interactions between sphingolipid receptors and their ligands. See, for example, Parker et al., (2000) Development of high throughput screening assays using fluorescence polarization: nuclear receptor-ligand-binding and kinase/phosphatase assays, J
Biomol Screen 5:77-88.

[0299] Fluorophores derived from sphingolipids that may be used in FP assays are commercially available. For example, Molecular Probes (Eugene, OR) currently sells sphingomyelin and one ceramide flurophores. These are, respectively, N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene- 3-pentanoyl)sphingosyl phosphocholine (BODIPY
FL C5-sphingomyelin); N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene- 3-dodecanoyl)sphingosyl phosphocholine (BODIPY FL C12-sphingomyelin); and N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene- 3-pentanoyl)sphingosine (BODIPY FL
C5-ceramide). U.S. Patent No. 4,150,949, (Immunoassay for gentamicin), discloses fluorescein-labelled gentamicins, including fluoresceinthiocarbanyl gentamicin. Additional fluorophores may be prepared using methods well known to the skilled artisan.

[0300] Exemplary normal-and-polarized fluorescence readers include the POLARION
fluorescence polarization system (Tecan AG, Hombrechtikon, Switzerland).
General multiwell plate readers for other assays are available, such as the VERSAMAX reader and the SPECTRAMAX multiwell plate spectrophotometer (both from Molecular Devices).

[0301] Fluorescence resonance energy transfer (FRET) is another useful assay for detecting interaction and has been described. See, e.g., Heim et al., (1996) Curr. Biol.
6:178-182; Mitra et al., (1996) Gene 173:13-17; and Selvin et al., (1995)1Vleth. Enzymol.
246:300-345. FRET
detects the transfer of energy between two fluorescent substances in close proximity, having known excitation and emission wavelengths. As an example, a protein can be expressed as a fusion protein with green fluorescent protein (GFP). When two fluorescent proteins are in proximity, such as when a protein specifically interacts with a target molecule, the resonance energy can be transferred from one excited molecule to the other. As a result, the emission spectrum of the sample shifts, which can be measured by a fluorometer, such as a fMAX
multiwell fluorometer (Molecular Devices, Sunnyvale Calif.).

[0302] Scintillation proximity assay (SPA) is a particularly useful assay for detecting an interaction with the target molecule. SPA is widely used in the pharmaceutical industry and has been described (Hanselman et al., (1997) J. Lipid Res. 38:2365-2373; Kahl et al., (1996) Anal. Biochem. 243:282-283; Undenfriend et al., (1987) Anal. Biochem. 161:494-500). See also U.S. Patent Nos. 4,626,513 and 4,568,649, and European Patent No.
0,154,734. One commercially available system uses FLASHPLATE scintillant-coated plates (NEN
Life Science Products, Boston, MA).

[0303] The target molecule can be bound to the scintillator plates by a variety of well known means. Scintillant plates are available that are derivatized to bind to fusion proteins such as GST, His6 or Flag fusion proteins. Where the target molecule is a protein complex or a multimer, one protein or subunit can be attached to the plate first, then the other components of the complex added later under binding conditions, resulting in a bound complex.

[0304] In a typical SPA assay, the gene products in the expression pool will have been radiolabeled and added to the wells, and allowed to interact with the solid phase, which is the immobilized target molecule and scintillant coating in the wells. The assay can be measured iinmediately or allowed to reach equilibrium. Either way, when a radiolabel becomes sufficiently close to the scintillant coating, it produces a signal detectable by a device such as a TOPCOUNT NXT microplate scintillation counter (Packard BioScience Co., Meriden Conn.). If a radiolabeled expression product binds to the target molecule, the radiolabel remains in proximity to the scintillant long enough to produce a detectable signal.

[0305] In contrast, the labeled proteins that do not bind to the target molecule, or bind only briefly, will not remain near the scintillant long enough to produce a signal above background.
Any time spent near the scintillant caused by random Brownian motion will also not result in a significant amount of signal. Likewise, residual unincorporated radiolabel used during the expression step may be present, but will not generate significant signal because it will be in solution rather than interacting with the target molecule. These non-binding interactions will therefore cause a certain level of background signal that can be mathematically removed. If too many signals are obtained, salt or other modifiers can be added directly to the assay plates until the desired specificity is obtained (Nichols et al., (1998) Anal.
Bioch.ern. 257:112-119).

Assay Compounds and Molecular Scaffolds [0306] Preferred characteristics of a scaffold include being of low molecular weight (e.g., less than 350 Da, or from about 100 to about 350 daltons, or from about 150 to about 300 daltons). Preferably clog P of a scaffold is from -1 to 8, more preferably less than 6, 5, or 4, most preferably less than 3. In particular embodiments the clogP is in a range -1 to an upper limit of 2, 3, 4, 5, 6, or 8; or is in a range of 0 to an upper limit of 2,3, 4, 5, 6, or 8. Preferably the number of rotatable bonds is less than 5, more preferably less than 4.
Preferably the number of hydrogen bond donors and acceptors is below 6, more preferably below 5. An additional criterion that can be useful is a polar surface area of less than 5. Guidance that can be useful in identifying criteria for a particular application can be found in Lipinski et al., (1997) Advanced Drug Delivefy Reviews 23 3-25, which is hereby incorporated by reference in its entirety.

[0307] A scaffold may preferably bind to a given protein binding site in a configuration that causes substituent moieties of the scaffold to be situated in pockets of the protein binding site.
Also, possessing chemically tractable groups that can be chemically modified, particularly through synthetic reactions, to easily create a combinatorial library can be a preferred characteristic of the scaffold. Also preferred can be having positions on the scaffold to which other moieties can be attached, which do not interfere with binding of the scaffold to the protein(s) of interest but do cause the scaffold to achieve a desirable property, for example, active transport of the scaffold to cells and/or organs, enabling the scaffold to be attached to a chromatographic column to facilitate analysis, or another desirable property.
A molecular scaffold can bind to a target molecule with any affinity, such as binding at high affinity, moderate affinity, low affinity, very low affinity, or extremely low affinity.

[0308] Thus, the above criteria can be utilized to select many compounds for testing that have the desired attributes. Many compounds having the criteria described are available in the commercial market, and may be selected for assaying depending on the specific needs to which the methods are to be applied.

[0309] A"compound library" or "library" is a collection of different compounds having different chemical structures. A coinpound library is screenable, that is, the compound library members therein may be subject to screening assays. In preferred embodiments, the library members can have a molecular weight of from about 100 to about 350 daltons, or from about 150 to about 350 daltons. Examples of libraries are provided aove.

[0310] Libraries of the present invention can contain at least one compound than binds to the target molecule at low affinity. Libraries of candidate compounds can be assayed by many different assays, such as those described above, e.g., a fluorescence polarization assay.
Libraries may consist of chemically synthesized peptides, peptidomimetics, or arrays of combinatorial chemicals that are large or small, focused or nonfocused. By "focused" it is meant that the collection of compounds is prepared using the structure of previously characterized compounds and/or pharmacophores.

[0311] Compound libraries may contain molecules isolated from natural sources, artificially synthesized molecules, or molecules synthesized, isolated, or otherwise prepared in such a manner so as to have one or more moieties variable, e.g., moieties that are independently isolated or randomly synthesized. Types of molecules in compound libraries include but are not limited to organic compounds, polypeptides and nucleic acids as those terms are used herein, and derivatives, conjugates and mixtures thereof.

[0312] Compound libraries of the invention may be purchased on the commercial market or prepared or obtained by any means including, but not limited to, combinatorial chemistry techniques, fermentation methods, plant and cellular extraction procedures and the like (see, e.g., Cwirla et al., (1990) Biochemistry, 87, 6378-6382; Houghten et al., (1991) Nature, 354, 84-86; Lam et al., (1991) Nature, 354, 82-84; Brenner et al., (1992) Proc.
Natl. Acad. Sci.
USA, 89, 5381-5383; R. A. Houghten, (1993) Trends Cpenet., 9, 235-239; E. R.
Felder, (1994) Chimia, 48, 512-541; Gallop et al., (1994) J. Med. Chem., 37, 1233-1251;
Gordon et al., (1994) .I. Med. Chein., 37, 1385-1401; Carell et al., (1995) Chein. Biol., 3, 171-183; Madden et al., Perspectives in Drug Discovery and Design 2, 269-282; Lebl et al., (1995) Biopolymers, 37 177-198); small molecules asseinbled around a shared molecular structure;
collections of chemicals that have been assembled by various commercial and noncommercial groups, natural products; extracts of marine organisms, fungi, bacteria, and plants.

[0313] Preferred libraries can be prepared in a homogenous reaction mixture, and separation of unreacted reagents from members of the library is not required prior to screening. Although many combinatorial chemistry approaches are based on solid state chemistry, liquid phase combinatorial chemistry is capable of generating libraries (Sun CM., (1999) Recent advances in liquid-phase combinatorial chemistry, Combirzatorial Chemistfy & High Tlaroughput Screening. 2:299-318).

[0314] Libraries of a variety of types of molecules are prepared in order to obtain members therefrom having one or more preselected attributes that can be prepared by a variety of techniques, including but not limited to parallel array synthesis (Houghton, (2000) Annu Rev Pharnaacol Toxicol 40:273-82, Parallel array and mixture-based synthetic coinbinatorial chemistry; solution-phase combinatorial chemistry (Merritt, (1998) Comb Chem High Throughput Screen 1(2):57-72, Solution phase combinatorial chemistry, Coe et al., (1998-99) Mol Divers;4(1):31-8, Solution-phase combinatorial chemistry, Sun, (1999) Conab Chem High Throughput Screen 2(6):299-318, Recent advances in liquid-phase combinatorial chemistry);
synthesis on soluble polymer (Gravert et al., (1997) Curr Opin Chem Biol 1(1):107-13, Synthesis on soluble polymers: new reactions and the construction of small molecules); and the like. See, e.g., Dolle et al., (1999) JComb Chem 1(4):235-82, Comprehensive survey of cominatorial library synthesis: 1998. Freidinger RM., (1999) Nonpeptidic ligands for peptide and protein receptors, Current Opinion in Chemical Biology; and Kundu et al., Prog Drug Res;53:89-156, Combinatorial chemistry: polymer supported synthesis of peptide and non-peptide libraries). Compounds may be clinically tagged for ease of identification (Chabala, (1995) Curr Opin Biotechnol 6(6):633-9, Solid-phase combinatorial chemistry and novel tagging methods for identifying leads).

[0315] The coinbinatorial synthesis of carbohydrates and libraries containing oligosaccharides have been described (Schweizer et al., (1999) Curr Opin Chem Biol 3(3):291-8,' Combinatorial synthesis of carbohydrates). The synthesis of natural-product based compound libraries has been described (Wessjohann, (2000) Curr Opin Chem Biol 4(3):303-9, Synthesis of natural-product based compound libraries).

[0316] Libraries of nucleic acids are prepared by various techniques, including by way of non-limiting example the ones described herein, for the isolation of aptamers.
Libraries that include oligonucleotides and polyaminooligonucleotides (Markiewicz et al., (2000) Synthetic oligonucleotide combinatorial libraries and their applications, Farnaaco.
55:174-7) displayed on streptavidin magnetic beads are known. Nucleic acid libraries are known that can be coupled to parallel sampling and be deconvoluted without complex procedures such as automated mass spectrometry (Enjalbal C. Martinez J. Aubagnac JL, (2000) Mass spectrometry in combinatorial chemistry, Mass Spectrometiy Reviews. 19:139-61) and parallel tagging. (Perrin DM., Nucleic acids for recognition and catalysis: landmarks, limitations, and looking to the future, Combinatorial Chemistry & High Throughput ScNeening 3:243-69).
[0317] Peptidomimetics are identified using combinatorial chemistry and solid phase synthesis (Kim HO. Kahn M., (2000) A merger of rational drug design and combinatorial chemistry: development and application of peptide secondary structure mimetics, Combinatorial Chemistry & High Throughput Screening 3:167-83; al-Obeidi, (1998) Mol Biotechnol 9(3):205-23, Peptide and peptidomimetric libraries. Molecular diversity and drug design). The synthesis may be entirely random or based in part on a known polypeptide.
[0318] Polypeptide libraries can be prepared according to various techniques.
In brief, phage display techniques can be used to produce polypeptide ligands (Gram H., (1999) Phage display in proteolysis and signal transduction, Combinatorial Chemistry & High Throughput Screening. 2:19-28) that may be used as the basis for synthesis of peptidomimetics.
Polypeptides, constrained peptides, proteins, protein domains, antibodies, single chain antibody fragments, antibody fragments, and antibody combining regions are displayed on filamentous phage for selection.

[0319] Large libraries of individual variants of human single chain Fv antibodies have been produced. See, e.g., Siegel RW. Allen B. Pavlik P. Marks JD. Bradbury A., (2000) Mass spectral analysis of a protein complex using single-chain antibodies selected on a peptide target: applications to functional genomics, Journal of Molecular Biology 302:285-93; Poul MA. Becerril B. Nielsen UB. Morisson P. Marks JD.,(2000) Selection of tumor-specific intemalizing human antibodies from phage libraries. Source Journal of MoleculaY Biology.
301:1149-61; Amersdorfer P. Marks JD., (2001) Phage libraries for generation of anti-botulinum scFv antibodies, Methods in Molecular Biology. 145:219-40; Hughes-Jones NC.
Bye JM. Gorick BD. Marks JD. Ouwehand WH., (1999) Synthesis of Rh Fv phage-antibodies using VH and VL germline genes, British Journal ofllaematology. 105:811-6;
McCall AM.
Amoroso AR. Sautes C. Marks JD. Weiner LM., (1998) Characterization of anti-mouse Fc gamma RII single-chain Fv fragments derived from human phage display libraries, Immunotechnology. 4:71-87; Sheets MD. Amersdorfer P. Finnern R. Sargent P.
Lindquist E.
Schier R. Hemingsen G. Wong C. Gerhart JC. Marks JD. Lindquist E., (1998) Efficient construction of a large nonimmune phage antibody library: the production of high-affinity human single-chain antibodies to protein antigens (published erratum appears in Proc Natl Acad Sci USA 1999 96:795), Proc Natl Acad Sci USA 95:6157-62).

[0320] Focused or smart chemical and pharmacophore libraries can be designed with the help of sophisticated strategies involving computational chemistry (e.g., Kundu B.
Khare SK.
Rastogi SK., (1999) Combinatorial chemistry: polymer supported synthesis of peptide and non-peptide libraries, Progress in Drug Research 53:89-156) and the use of structure-based ligands using database searching and docking, de novo drug design and estimation of ligand binding affinities (Joseph-McCarthy D., (1999) Computational approaches to structure-based ligand design, Phaf naacology & Therapeutics 84:179-91; Kirkpatrick DL. Watson S. Ulhaq S., (1999) Structure-based drug design: combinatorial chemistry and molecular modeling, Coinbinatorial Claemistry & High Througlaput Screening. 2:211-21; Eliseev AV.
Lehn JM., (1999) Dynamic combinatorial chemistry: evolutionary formation and screening of molecular libraries, Current Topics in Microbiology & Immunology 243:159-72; Bolger et al., (1991) Methods Enz. 203:21-45; Martin, (1991) Methods Enz. 203:587-613; Neidle et al., (1991) Methods Enz. 203:433-458; U.S. Patent 6,178,384).

X. Crystallography [0321] After binding compounds have been determined, the orientation of compound bound to target is determined. Preferably this determination involves crystallography on co-crystals of molecular scaffold compounds with target. Most protein crystallographic platforms can preferably be designed to analyze up to about 500 co-coinplexes of compounds, ligands, or molecular scaffolds bound to protein targets due to the physical parasneters of the instruxnents and convenience of operation. If the number of scaffolds that have binding activity exceeds a number convenient for the application of crystallography methods, the scaffolds can be placed into groups based on having at least one common chemical structure or other desirable characteristics, and representative compounds can be selected from one or more of the classes.
Classes can be made witli increasingly exacting criteria until a desired number of classes (e.g., 500) is obtained. The classes can be based on chemical structure similarities between molecular scaffolds in the class, e.g., all possess a pyrrole ring, benzene ring, or other chemical feature. Likewise, classes can be based on shape characteristics, e.g., space-filling characteristics.

[0322] The co-crystallography analysis can be performed by co-complexing each scaffold with its target at concentrations of the scaffold that showed activity in the screening assay.
This co-complexing can be accomplished with the use of low percentage organic solvents with the target molecule and then concentrating the target with each of the scaffolds. In preferred embodiments these solvents are less than 5% organic solvent such as dimethyl sulfoxide (DMSO), ethanol, metlzanol, or ethylene glycol in water or another aqueous solvent. Each scaffold complexed to the target molecule can then be screened with a suitable number of crystallization screening conditions at both 4 and 20 degrees. In preferred embodiments, about 96 crystallization screening conditions can be performed in order to obtain sufficient information about the co-complexation and crystallization conditions, and the orientation of the scaffold at the binding site of the target molecule. Crystal structures can then be analyzed to determine how the bound scaffold is oriented physically within the binding site or within one or more binding pockets of the molecular family member.

[0323] It is desirable to detennine the atomic coordinates of the compounds bound to the target proteins in order to determine which is a most suitable scaffold for the protein family.
X-ray crystallographic analysis is tlierefore most preferable for determining the atomic coordinates. Those compounds selected can be fur-ther tested witll the application of medicinal chemistry. Compounds can be selected for medicinal chemistry testing based on their binding position in the target molecule. For example, when the compound binds at a binding, site, the compound's binding position in the binding site of the target molecule can be considered with respect to the chemistry that can be performed on chemically tractable structures or sub-structures of the compound, and how such modifications on the compound might interact with structures or sub-structures on the binding site of the target. Thus, one can explore the binding site of the target and the chemistry of the scaffold in order to make decisions on how to modify the scaffold to arrive at a ligand with higher potency and/or selectivity.
This process allows for more direct design of ligands, by utilizing structural and chemical information obtained directly from the co-complex, thereby enabling one to more efficiently and quickly design lead compounds that are likely to lead to beneficial drug products. In various embodiments it may be desirable to perform co-crystallography on all scaffolds that bind, or only those that bind with a particular affinity, for example, only those that bind with high affinity, moderate affinity, low affinity, very low affinity, or extremely low affinity. It may also be advantageous to perform co-crystallography on a selection of scaffolds that bind with any combination of affinities.

[0324] Standard X-ray protein diffraction studies such as by using a Rigaku RU-(Rigaku, Tokyo, Japan) with an X-ray imaging plate detector or a synchrotron beam-line can be performed on co-crystals and the diffraction data measured on a standard X-ray detector, such as a CCD detector or an X-ray imaging plate detector.

[0325] Performing X-ray crystallography on about 200 co-crystals should generally lead to about 50 co-crystals structures, which should provide about 10 scaffolds for validation in chemistry, which should finally result in about 5 selective leads for target molecules.

Virtual Assays [0326] Commercially available software that generates three-dimensional graphical representations of the complexed target and compound from a set of coordinates provided can be used to illustrate and study how a compound is oriented when bound to a target. (e.g., QUANTA , Accelerys, San Diego, CA). Thus, the existence of binding pockets at the binding site of the targets can be particularly useful in the present invention. These binding pockets are revealed by the crystallographic structure determination and show the precise chemical interactions involved in binding the compound to the binding site of the target. The person of ordinary skill will realize that the illustrations can also be used to decide where chemical groups might be added, substituted, modified, or deleted from the scaffold to enhance binding or another desirable effect, by considering where unoccupied space is located in the complex and which chemical substructures might have suitable size and/or charge characteristics to fill it. The person of ordinary skill will also realize that regions within the binding site can be flexible and its properties can change as a result of scaffold binding, and that chemical groups can be specifically targeted to those regions to achieve a desired effect.
Specific locations on the molecular scaffold can be considered with reference to where a suitable chemical substructure can be attached and in which conformation, and which site has the most advantageous chemistry available.

[0327] An understanding of the forces that bind the compounds to the target proteins reveals which compounds can most advantageously be used as scaffolds, and which properties can most effectively be manipulated in the design of ligands. The person of ordinary skill will realize that steric, ionic, hydrogen bond, and other forces can be considered for their contribution to the maintenance or enhancement of the target-compound complex.
Additional data can be obtained with automated computational methods, such as docking and/or Free Energy Perturbations (FEP), to account for other energetic effects such as desolvation penalties. The compounds selected can be used to generate information about the chemical interactions with the target or for elucidating chemical modifications that can enhance selectivity of binding of the compound.

[0328] Computer models, such as homology models (i.e., based on a known, experimentally derived structure) can be constructed using data from the co-crystal structures. When the target molecule is a protein or enzyme, preferred co-crystal structures for making homology models contain high sequence identity in the binding site of the protein sequence being modeled, and the proteins will preferentially also be within the same class and/or fold family.
Knowledge of conserved residues in active sites of a protein class can be used to select homology models that accurately represent the binding site. Homology models can also be used to map structural information from a surrogate protein where an apo or co-crystal structure exists to the target protein.

[0329] Virtual screening methods, such as docking, can also be used to predict the binding configuration and affinity of scaffolds, compounds, and/or combinatorial library members to homology models. Using this data, and carrying out "virtual experiments" using computer software can save substantial resources and allow the person of ordinary skill to make decisions about which compounds can be suitable scaffolds or ligands, without having to actually synthesize the ligand and perform co-crystallization. Decisions thus can be made about which compounds merit actual synthesis and co-crystallization. An understanding of such chemical interactions aids in the discovery and design of drugs that interact more advantageously with target proteins and/or are more selective for one protein family member over others. Thus, applying these principles, compounds with superior properties can be discovered.

[0330] Additives that promote co-crystallization can of course be included in the target molecule formulation in order to enhance the formation of co-crystals. In the case of proteins or enzymes, the scaffold to be tested can be added to the protein formulation, which is preferably present at a concentration of approximately 1 mg/ml. The formulation can also cointain between 0%-10% (v/v) organic solvent, e.g. DMSO, methanol, ethanol, propane diol, or 1,3 dimethyl propane diol (MPD) or some combination of those organic solvents.
Compounds are preferably solubilized in the organic solvent at a concentration of about 10 mM and added to the protein sample at a concentration of about 100 mM. The protein-compound complex is then concentrated to a final concentration of protein of from about 5 to about 20 mg/ml. The complexation and concentration steps can conveniently be performed using a 96-well formatted concentration apparatus (e.g., Amicon Inc., Piscataway, NJ).
Buffers and other reagents present in the formulation being crystallized can contain other components that promote crystallization or are compatible with crystallization conditions, such as DTT, propane diol, glycerol.

[0331] The crystallization experiment can be set-up by placing small aliquots of the concentrated protein-compound complex (1 l) in a 96 well format and sampling under 96 crystallization conditions. (Other screening formats can also be used, e.g., plates with greater than 96 wells.) Crystals can typically be obtained using standard crystallization protocols that can involve the 96 well crystallization plate being placed at different temperatures. Co-crystallization varying factors other than temperature can also be considered for each protein-compound coiuplex if desirable. For example, atmospheric pressure, the presence or absence of light or oxygen, a change in gravity, and many other variables can all be tested. The person of ordinary skill in the art will realize other variables that can advantageously be varied and considered.

Ligand Design and Preparation [0332] The design and preparation of ligands can be performed with or without structural and/or co-crystallization data by considering the chemical structures in common between the active scaffolds of a set. In this process structure-activity hypotheses can be formed and those chemical structures found to be present in a substantial number of the scaffolds, including those that bind with low affinity, cau be presumed to have some effect on the binding of the scaffold. This binding can be presumed to induce a desired biochemical effect when it occurs in a biological system (e.g., a treated mammal). New or modified scaffolds or combinatorial libraries derived from scaffolds can be tested to disprove the maximum number of binding and/or structure-activity hypotheses. The remaining hypotheses can then be used to design ligands that achieve a desired binding and biochemical effect.

[0333] But in many cases it will be preferred to have co-crystallography data for consideration of how to modify the scaffold to achieve the desired binding effect (e.g., binding at higher affinity or with higher selectivity). Using the case of proteins and enzymes, co-crystallography data shows the binding pocket of the protein with the molecular scaffold bound to the binding site, and it will be apparent that a modification can be made to a chemically tractable group on the scaffold. For example, a small volume of space at a protein binding site or pocket might be filled by modifying the scaffold to include a small chemical group that fills the volume. Filling the void volume can be expected to result in a greater binding affinity, or the loss of undesirable binding to another member of the protein family.
Similarly, the co-crystallography data may show that deletion of a chemical group on the scaffold may decrease a hindrance to binding and result in greater binding affinity or specificity.

[0334] It can be desirable to take advantage of the presence of a charged chemical group located at the binding site or pocket of the protein. For example, a positively charged group can be complemented with a negatively charged group introduced on the molecular scaffold.
This can be expected to increase binding affinity or binding specificity, thereby resulting in a more desirable ligand. In many cases, regions of protein binding sites or pockets are known to vary from one family member to another based on the amino acid differences in those regions.
Chemical additions in such regions can result in the creation or elimination of certain interactions (e.g., hydrophobic, electrostatic, or entropic) that allow a compound to be more specific for one protein target over another or to bind with greater affinity, thereby enabling one to synthesize a compound with greater selectivity or affinity for a particular family member. Additionally, certain regions can contain amino acids that are known to be more flexible than others. This often occurs in amino acids contained in loops connecting elements of the secondary structure of the protein, such as alpha helices or beta strands. Additions of chemical moieties can also be directed to these flexible regions in order to increase the likelihood of a specific interaction occurring between the protein target of interest and the compotuid. Virtual screening methods can also be conducted in silico to assess the effect of chemical additions, subtractions, modifications, and/or substitutions on compounds with respect to members of a protein family or class.

[0335] The addition, subtraction, or modification of a chemical structure or sub-structure to a scaffold can be performed with any suitable chemical moiety. For example the following moieties, which are provided by way of example and are not intended to be limiting, can be utilized: hydrogen, alkyl, alkoxy, phenoxy, alkenyl, alkynyl, phenylalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, alkyloxy, alkylthio, alkenylthio, phenyl, phenylalkyl, phenylalkylthio, hydroxyalkyl-thio, alkylthiocarbbamylthio, cyclohexyl, pyridyl, piperidinyl, alkylamino, amino, nitro, mercapto, cyano, hydroxyl, a halogen atom, halomethyl, an oxygen atom (e.g., forming a ketone or N-oxide) or a sulphur atom (e.g., forming a thiol, thione, di-alkylsulfoxide or sulfone) are all examples of moieties that can be utilized.

[0336] Additional examples of structures or sub-structures that may be utilized are an aryl optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, carboxamide, nitro, and ester moieties; an amine of formula -NX2X3, where X2 and X3 are independently selected from the group consisting of hydrogen, saturated or unsaturated alkyl, and homocyclic or heterocyclic ring moieties; halogen or trihalomethyl; a ketone of formula -COX4, where X4 is selected from the group consisting of alkyl and homocyclic or heterocyclic ring moieties; a carboxylic acid of formula -(X5)õCOOH or ester of formula (X6)õCOOX7, where X5, X6, and X7 and are independently selected from the group consisting of alkyl and homocyclic or heterocyclic ring moieties and where n is 0 or 1; an alcohol of formula (X8)õOH or an alkoxy moiety of formula -(X$)nOX9, where X8 and X9 are independently selected from the group consisting of saturated or unsaturated alkyl and homocyclic or heterocyclic ring moieties, wherein said ring is optionally substituted with one or more substituents independently selected .from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, nitro, and ester and where n is 0 or 1; an amide of formula NHCOXio, where Xlo is selected from the group consisting of alkyl, hydroxyl, and homocyclic or heterocyclic ring moieties, wherein said ring is optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, nitro, and ester; SO2, NXII
X12, where Xl l and X12 are selected from the group consisting of hydrogen, alkyl, and homocyclic or heterocyclic ring moieties; a homocyclic or heterocyclic ring moiety optionally substituted witli one, two, or three substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, carboxamide, nitro, and ester moieties; an aldehyde of formula -CHO; a sulfone of formula -S02X13, where X13 is selected from the group consisting of saturated or unsaturated alkyl and homocyclic or heterocyclic ring moieties; and a nitro of formula -NO2.

Identification of Attachment Sites on Molecular Scaffolds and Ligands [0337] In addition to the identification and development of ligands for phosphodiesterases and other enzymes, determination of the orientation of a molecular scaffold or other binding compound in a binding site allows identification of energetically allowed sites for attachment of the binding molecule to another component. For such sites, any free energy change associated with the presence of the attached component should not destablize the binding of the compound to the phosphodiesterase to an extent that will disrupt the binding.
Preferably, the binding energy with the attachment should be at least 4 kcal/mol., more preferably at least 6, 8, 10, 12, 15, or 20 kcal/mol. Preferably, the presence of the attachment at the particular site reduces binding energy by no more than 3, 4, 5, 8, 10, 12, or 15 kcal/mol.

[0338] In many cases, suitable attachment sites will be those that are exposed to solvent when the binding compound is bound in the binding site. In some cases, attachment sites can be used that will result in small displacements of a portion of the enzyme without an excessive energetic cost. Exposed sites can be identified in various ways. For example, exposed sites can be identified using a graphic display or 3-dimensional model. In a grahic display, such as a computer display, an image of a compound bound in a binding site can be visually inspected to reveal atoms or groups on the compound that are exposed to solvent and oriented such that attachment at such atom or group would not preclude binding of the enzyme and binding compound. Energetic costs of attachment can be calculated based on changes or distortions that would be caused by the attachment as well as entropic changes.

[0339] Many different types of components can be attached. Persons with skill are familiar with the chemistries used for various attachments. Examples of components that can be attached include, without limitation: solid phase components such as beads, plates, chips, and wells; a dlrect or indirect label; a linker, which may be a traceless linker;
among others. Such linkers can themselves be attached to other components, e.g., to solid phase media, labels, and/or binding moieties.

[0340] The binding energy of a compound and the effects on binding energy for attaching the molecule to another component can be calculated approximately using any of a variety of available software or by manual calculation. An example is the following:

[0341] Calculations were performed to estimate binding energies of different organic molecules to two Kinases: PIM-1 and CDK2. The organic molecules considered included Staurosporine, identified compounds that bind to PDE5A, and several linkers.

[0342] Calculated binding energies between protein-ligand complexes were obtained using the FlexX score (an iinplementation of the Bolun scoring function) within the Tripos software suite. The form for that equation is shown in the equation below:

OGbind = AGtr + AGhb + AGion + AGlipo + OGarom + AGrot wherein: AGtr is a constant term that accounts for the overall loss of rotational and translational entropy of the lignand, AGhb accounts for hydrogen bonds formed between the ligand and protein, AGion accounts for the ionic interactions between the ligand and protein, AGlipo accounts for the lipophilic interaction that corresponds to the protein-ligand contact surface, OGarom accounts for interactions between aromatic rings in the protein and ligand, and AGrot accounts for the entropic penalty of restricting rotatable bonds in the ligand upon binding.

[0343] This method estimates the free energy that a lead compound should have to a target protein for which there is a crystal structure, and it accounts for the entropic penalty of flexible linkers. It can therefore be used to estimate the free energy penalty incurred by attaching linkers to molecules being screened and the binding energy that a lead compound should have in order to overcome the free energy penalty of the linker. The method does not account for solvation and the entropic penalty is likely overestimated for cases where the linker is bound to a solid phase through another binding complex, such as a biotin:streptavidin complex.

[0344] Co-crystals were aligned by superimposing residues of PIM-1 with corresponding residues in CDK2. The PIM-1 structure used for these calculations was a co-crystal of PIM-1 with a binding coinpound. The CDK2:Staurosporine co-crystal used was from the Brookhaven database file laql. Hydrogen atoms were added to the proteins and atomic charges were assigned using the AMBER95 parameters within Sybyl. Modifications to the compounds described were made within the Sybyl modeling suite from Tripos.

[0345] These calcualtions indicate that the calculated binding energy for compounds that bind strongly to a given target (such as Staurosporine:CDK2) can be lower than -25 kcal/mol, while the calculated binding affinity for a good scaffold or an unoptimized binding compound can be in the range of -15 to -20. The free energy penalty for attachment to a linker such as the ethylene glycol or hexatriene is estimated as typically being in the range of +5 to + 15 kcal/mol.
Linkers [0346] Linkers suitable for use in the invention can be of many different types. Linkers can be selected for particular applications based on factors such as linker chemistry compatible for attachment to a binding compound and to another component utilized in the particular application. Additional factors can include, without limitation, linker length, linker stability, and ability to remove the linker at an appropriate time. Exemplary linkers include, but are not limited to, hexyl, hexatrienyl, ethylene glycol, and peptide linkers.
Traceless linkers can also be used, e.g., as described in Plunkett, M. J., and Ellman, J. A., (1995), J.
Org. Chem., 60:6006.
[0347] Typical functional groups, that are utilized to link binding compound(s), include, but not limited to, carboxylic acid, amine, hydroxyl, and thiol. (Examples can be found in Solid-supported combinatorial and parallel synthesis of small molecular weight compound libraries;
(1998) Tetrahedron organic chemistry series Vol.17; Pergamon; p85).

Labels [0348] As indicated above, labels can also be attached to a binding compound or to a linker attached to a binding compound. Such attachment may be direct (attached directly to the binding compound) or indirect (attached to a component that is directly or indirectly attached to the binding compound). Such labels allow detection of the compound either directly or indirectly. Attachement of labels can be performed using conventional chemistries. Labels can include, for example, fluorescent labels, radiolabels, light scattering particles, light absorbent particles, magnetic particles, enzymes, and specific binding agents (e.g., biotin or an antibody target moiety).

Solid Phase Media [0349] Additional examples of components that can be attached directly or indirectly to a binding compound include various solid phase media. Similar to attachment of linkers and labels, attachment to solid phase media can be performed using conventional chemistries. Such solid phase media can include, for example, small components such as beads, nanoparticles, and fibers (e.g., in suspension or in a gel or chromatographic matrix).
Likewise, solid phase media can include larger objects such as plates, chips, slides, and tubes. In many cases, the binding compound will be attached in only a portion of such an objects, e.g., in a spot or other local element on a generally flat surface or in a well or portion of a well.

Identification of Biological Agents [0350] The posession of structural information about a protein also provides for the identification of useful biological agents, such as epitpose for development of antibodies, identification of mutation sites expected to affect activity, and identification of attachment sites allowing attachment of the protein to materials such as labels, linkers, peptides, and solid phase media.

[0351] Antibodies (Abs) finds multiple applications in a variety of areas including biotechnology, medicine and diagnosis, and indeed they are one of the most powerful tools for life science research. Abs directed against protein antigens can recognize either linear or native three-dimensional (3D) epitopes. The obtention of Abs that recognize 3D
epitopes require the use of whole native protein (or of a portion that assumes a native conformation) as immunogens. Unfortunately, this not always a choice due to various technical reasons: for example the native protein is just not available, the protein is toxic, or its is desirable to utilize a high density antigen presentation. In such cases, immunization with peptides is the alternative. Of course, Abs generated in this manner will recognize linear epitopes, and they might or might not recognize the source native protein, but yet they will be useful for standard laboratory applications such as western blots. The selection of peptides to use as immunogens can be accomplished by following particular selection rules and/or use of epitope prediction software.

[0352] Though methods to predict antigenic peptides are not infallible, there are several rules that can be followed to determine what peptide fragments from a protein are likely to be antigenic. These rules are also dictated to increase the likelihood that an Ab to a particular peptide will recognize the native protein.

= 1. Antigenic peptides should be located in solvent accessible regions and contain both hydrophobic and hydrophilic residues.

o For proteins of known 3D structure, solvent accessibility can be determined using a variety of programs such as DSSP, NACESS, or WHATIF, among others.

o If the 3D structure is not known, use any of the following web servers to predict accessibilities: PHD, JPRED, PredAcc (c) ACCpro = 2. Preferably select peptides lying in long loops connecting Secondary Structure (SS) motifs, avoiding peptides located in helical regions. This will increase the odds that the Ab recognizes the native protein. Such peptides can, for example, be identified from a crystal structure or crystal structure-based homology model.

o For protein with known 3D coordinates, SS can be obtained from the sequence link of the relevant entry at the Brookhaven data bank. The PDBsum server also offer SS analysis of pdb records.

o When no structure is available secondary structure predictions can be obtained from any of the following servers: PHD, JPRED, PSI-PRED, NNSP, etc = 3. When possible, choose peptides that are in the N- and C-terminal region of the protein. Because the N- and C- terminal regions of proteins are usually solvent accessible and unstructured, Abs against those regions are also likely to recognize the native protein.

= 4. For cell surface glycoproteins, eliminate from initial peptides those containing consesus sites for N-glycosilation.

o N-glycosilation sites can be detected using Scanprosite, or NetNGlyc [0353] In addition, several methods based on various physio-chemical properties of experimental determined epitopes (flexibility, hydrophibility, accessibility) have been published for the prediction of antigenic determinants and can be used. The antigenic index and Preditop are example.

[0354] Perhaps the simplest method for the prediction of antigenic determinants is that of Kolaskar and Tongaonkar, which is based on the occurrence of amino acid residues in experimentally determined epitopes. (Kolaskar and Tongaonkar (1990) A semi-empirical method for prediction of antigenic determinants on protein antigens. FEBBS
Lett. 276(1-2):172-174.) The prediction algorithm works as follows:

= 1. Calculate the average propensity for each overlapping 7-mer and assign the result to the central residue (i+3) of the 7-mer.

= 2. Calculate the average for the whole protein.

= 3. (a) If the average for the whole protein is above 1.0 then all residues having average propensity above 1.0 are potentially antigenic.

= 4. (b) If the average for the whole protein is below 1.0 then all residues having above the average for the whole protein are potentially antigenic.

= 5. Find 8-mers where all residues are selected by step 3 above (6-mers in the original paper) [0355] The Kolaskar and Tongaonkar method is also available from the GCG
package, and it runs using the command egcg.

[0356] Crystal structures also allow identification of residues at which mutation is likely to alter the activity of the protein. Such residues include, for example, residues that interact with susbtrate, conserved active site residues, and residues that are in a region of ordered secondary structure of involved in tertiary interactions. The mutations that are likely to affect activity will vary for different molecular contexts. Mutations in an active site that will affect activity are typically substitutions or deletions that eliininate a charge-charge or hydrogen bonding interaction, or introduce a steric interference. Mutations in secondary structure regions or molecular interaction regions that are likely to affect activity include, for example, substitutions that alter the hydrophobicity/hydrophilicity of a region, or that introduce a sufficient strain in a region near or including the active site so that critical residue(s) in the active site are displaced. Such substitutions and/or deletions and/or insertions are recognized, and the predicted structural and/or energetic effects of mutations can be calculated using conventional software.

XI. Phosphodiesterase Activity Assays [0357] A number of different assays for phosphodiesterase activity can be utilized for assaying for active modulators and/or determining specificity of a modulator for a particular phosphodiesterase or group or phosphodiesterases. In addition to the assay mentioned in the Examples below, one of ordinary skill in the art will know of other assays that can be utilized and can modify an assay for a particular application. For example, numerous papers concerning PDEs described assays that can be used. For exainple, useful assays are described in Fryburg et al., U.S. Patent Application Publication 2002/0165237, Thompson et al., U.S.
Patent Application Publication 2002/0009764, Pamukcu et al., U.S. Patent Application 09/046,739, and Pamukcu et al., U.S. Patent 6,500,610.

[0358] An assay for phosphodiesterase activity that can be used for PDE4B, can be performed according to the following procedure using purified PDE4B using the procedure described in the Examples.

[0359] Additional alternative assays can employ binding determinations. For example, this sort of assay can be formatted either in a fluorescence resonance energy transfer (FRET) format, or using an AlphaScreen (amplified luminescent proximity homogeneous assay) fonnat by varying the donor and acceptor reagents that are attached to streptavidin or the phosphor-specific antibody.

XII. Organic Synthetic Techniques [0360] The versatility of computer-based modulator design and identification lies in the diversity of structures screened by the computer programs. The computer programs can search databases that contain very large numbers of molecules and can modify modulators already complexed with the enzyme with a wide variety of chemical functional groups. A
consequence of this chemical diversity is that a potential modulator of phosphodiesterase function may take a chemical fonn that is not predictable. A wide array of organic synthetic techniques exist in the art to meet the challenge of constructing these potential modulators.
Many of these organic synthetic methods are described in detail in standard reference sources utilized by those skilled in the art. One example of suh a reference is March, 1994, Advanced Organic Chemistry; Reactions, Mechanisms and Structure, New York, McGraw Hill.
Thus, the teclmiques useful to synthesize a potential modulator of phosphodiesterase function identified by computer-based methods are readily available to those skilled in the art of organic chemical synthesis.

XIII. Isomers, Prodrugs, and Active Metabolites [0361] The present compounds are described herein with generic formulas and specific compounds. In addition, the present compounds may exist in a number of different fonns or derivatives, all within the scope of the present invention. These include, for example, tautomers, enantiomers, stereoisomers, racemic mixtures, regioisomers, salts, prodrugs (e.g., carboxylic acid esters), solvated forms, different crystal forms or polymorphs, and active metabolites A. Tautomers, Stereoisomers, Regioisomers, and Solvated Forms [0362] It is understood that certain compounds may exhibit tautomerism. In such cases, the formula drawings within this specification expressly depict only one of the possible tautomeric forms. It is therefore to be understood that within the invention the formulas are intended to represent any tautomeric form of the depicted compounds and are not to be limited merely to the specific tautomeric form depicted by the formula drawings.

[0363] Likewise, some of the present coinpounds may contain one or more chiral centers, and therefore, may exist in two or more stereoisomeric forms. Thus, such compounds may be present as single stereoisomers (i.e., essentially free of other stereoisomers), racemates, and/or mixtures of enantiomers and/or diastereomers. All such single stereoisomers, racemates and mixtures thereof are intended to be within the scope of the present invention.
Unless specified to the contrary, all such steroisomeric forms are included within the formulas provided herein.

[0364] In certain embodiments, a chiral compound of the present invention is in a form that contains at least 80% of a single isomer (60% enantiomeric excess ("e.e.") or diastereomeric excess ("d.e.")), or at least 85% (70% e.e. or d.e.), 90% (80% e.e. or d.e.), 95% (90% e.e. or d.e.), 97.5% (95% e.e. or d.e.), or 99% (98% e.e. or d.e.). As generally understood by those skilled in the art, an optically pure compound having one chiral center is one that consists essentially of one of the two possible enantiomers (i.e., is enantiomerically pure), and an optically pure compound having more than one chiral center is one that is both diastereomerically pure and enantiomerically pure. In certain embodiments, the compound is present in optically pure form.

[0365] For compounds is which synthesis involves addition of a single group at a double bond, particularly a carbon-carbon double bond, the addition may occur at either of the double bond-linked atoms. For such compounds, the present invention includes both such regioisomers.

[0366] Additionally, the formulas are intended to cover solvated as well as unsolvated forms of the identified structures. For example, the indicated structures include both both hydrated and non-hydrated forms. Other examples of solvates include the structures in combination with isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, or ethanolamine.

B. Prodrugs and Metabolites [0367] In addition to the present formulas and compounds described herein, the invention also includes prodrugs (generally pharmaceutically acceptable prodrugs), active metabolic derivatives (active metabolites), and their pharmaceutically acceptable salts.

[0368] In this context, prodrugs are compounds that may be converted under physiological conditions or by solvolysis to the specified compound or to a pharmaceutically acceptable salt of such a compound. A common example is an alkyl ester of a carboxylic acid.

[0369] As described in The Practice of Medicinal Chemistry, Ch. 31-32 (Ed.
Wermuth, Academic Press, San Diego, CA, 2001), prodrugs can be conceptually divided into two non-exclusive categories, bioprecursor prodrugs and carrier prodrugs. Generally, bioprecursor prodrugs are compounds are inactive or have low activity compared to the corresponding active drug compound, that contain one or more protective groups and are converted to an active form by metabolism or solvolysis. Both the active drug form and any released metabolic products should have acceptably low toxicity. Typically, the formation of active drug compound involves a metabolic process or reaction that is one of the follow types:
[0370] Oxidative reactions, such as oxidation of alcohol, carbonyl, and acid functions, hydroxylation of aliphatic carbons, hydroxylation of alicyclic carbon atoms, oxidation of aromatic carbon atoms, oxidation of carbon-carbon double bonds, oxidation of nitrogen-containing functional groups, oxidation of silicon, phosphorus, arsenic, and sulfur, oxidative N-delakylation, oxidative 0- and S-delakylation, oxidative deamination, as well as other oxidative reactions.

[0371] Reductive reactions, such as reduction of carbonyl groups, reduction of alcoholic groups and carbon-carbon double bonds, reduction of nitrogen-containing functions groups, and other reduction reactions.

[0372] Reactions without change in the state of oxidation, such as liydrolysis of esters and ethers, hydrolytic cleavage of carbon-nitrogen single bonds, hydrolytic cleavage of non-aromatic heterocycles, hydration and dehydration at multiple bonds, new atomic linkages resulting from dehydration reactions, hydrolytic dehalogenation, removal of hydrogen halide molecule, and other such reactions.

[0373] Carrier prodrugs are drug compounds that contain a transport moiety, e.g., that improves uptake and/or localized delivery to a site(s) of action. Desirably for such a carrier prodrug, the linkage between the drug moiety and the transport moiety is a covalent bond, the prodrug is inactive or less active than the drug compound, the prodrug and any release transport moiety are acceptably non-toxic. For prodrugs where the transport moiety in intended to enhance uptake, typically the release of the transport moiety should be rapid. In other cases, it is desirable to utilize a moiety that provides slow release, e.g., certain polymers or other moieties, such as cyclodextrins. (See, e.g., Cheng et al., U.S.
Patent publ.
20040077595, appl. 10/656,838, incorporated herein by reference.) Such carrier prodrugs are often advantageous for orally administered drugs. Carrier prodrugs can, for example, be used to improve one or more of the following properties: increased lipophilicity, increased duration of pharmacological effects, increased site-specificity, decreased toxicity and adverse reactions, and/or improvement in drug formulation (e.g., stability, water solubility, suppression of an undesirable organoleptic or physiochemical property). For example, lipophilicity can be increased by esterification of hydroxyl groups with lipophilic carboxylic acids, or of carboxylic acid groups with alcohols, e.g., aliphatic alcohols. Wermuth, The Practice of Medicinal Chemistry, Ch. 31-32, Ed. Wermuth, Academic Press, San Diego, CA, 2001.

[0374] Prodrugs may proceed from prodrug form to active forin in a single step or may have one or more intermediate forms which may themselves have activity or may be inactive.
[0375] Metabolites, e.g., active metabolites overlap with prodrugs as described above, e.g., bioprecursor prodrugs. Thus, such metabolites are pharmacologically active compounds or compounds that further metabolize to pharmacologically active compounds that are derivatives resulting from metabolic process in the body of a subject or patient. Of these, active metabolites are such pharmacologically active derivative compounds. For prodrugs, the prodrug compounds is generally inactive or of lower activity than the metabolic product. For active metabolites, the parent compound may be either an active compound or may be an inactive prodrug.

[0376] Prodrugs and active metabolites may be identified using routine techniques know in the art. See, e.g., Bertolini et al, 1997, JMed Claern 40:2011-2016; Shan et al., JPharnz Sci 86:756-757; Bagshawe, 1995, Drug Dev Res 34:220-230; Wermuth, The Practice of Medicinal Chemistry, Ch. 31-32, Academic Press, San Diego, CA, 2001.

C. Pharmaceutically acceptable salts [0377] Compounds can be formulated as or be in the form of pharmaceutically acceptable salts. Pharmaceutically acceptable salts are non-toxic salts in the amounts and concentrations at which they are administered. The preparation of such salts can facilitate the pharmacological use by altering the physical characteristics of a compound without preventing it from exerting its physiological effect. Useful alterations in physical properties include lowering the melting point to facilitate transmucosal administration and increasing the solubility to facilitate administering higher concentrations of the drug.

[0378] Pharmaceutically acceptable salts include acid addition salts such as those containing sulfate, chloride, hydrochloride, f-umarate, maleate, phosphate, sulfamate, acetate, citrate, lactate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, cyclohexylsulfamate and quinate. Phannaceutically acceptable salts can be obtained from acids such as hydrochloric acid, maleic acid, sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic acid, fumaric acid, and quinic acid.

[0379] Pharmaceutically acceptable salts also include basic addition salts such as those containing benzathine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, ammonium, alkylamine, and zinc, when acidic functional groups, such as carboxylic acid or phenol are present. For example, see Remington's Phannaceutical Sciences, 19th ed., Mack Publishing Co., Easton, PA, Vol. 2, p. 1457, 1995. Such salts can be prepared using the appropriate corresponding bases.

[0380] Pharmaceutically acceptable salts can be prepared by standard techniques. For example, the free-base form of a compound is dissolved in a suitable solvent, such as an aqueous or aqueous-alcohol in solution containing the appropriate acid and then isolated by evaporating the solution. In another example, a salt is prepared by reacting the free base and acid in an organic solvent.

[0381] Thus, for example, if the particular compound is a base, the desired pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, or with an organic acid, such as acetic acid, maleic acid, succinic acid, mandelic acid, fiimaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid, such as glucuronic acid or galacturonic acid, an alpha-hydroxy acid, such as citric acid or tartaric acid, an amino acid, such as aspartic acid or glutamic acid, an aromatic acid, such as benzoic acid or cinnamic acid, a sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid, or the like.

[0382] Similarly, if the particular compound is an acid, the desired pharmaceutically acceptable salt may be prepared by any suitable method, for example, treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary or tertiary), an alkali metal hydroxide or alkaline earth metal hydroxide, or the like. Illustrative examples of suitable salts include organic salts derived from amino acids, such as glycine and arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines, such as piperidine, morpholine and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.

[0383] The phannaceutically acceptable salt of the different compounds may be present as a complex. Examples of complexes include 8-chlorotheophylline complex (analogous to, e.g., dimenhydrinate: diphenhydramine 8-chlorotheophylline (1:1) complex; Dramamine) and various cyclodextrin inclusion complexes.

[0384] Unless specified to the contrary, specification of a compound herein includes pharmaceutically acceptable salts of such compound.

D. Polymorphic forms [0385] In the case of agents that are solids, it is understood by those skilled in the art that the compounds and salts may exist in different crystal or polyinorphic forms, all of which are intended to be within the scope of the present invention and specified fonnulas.

XIV. Administration [0386] The methods a.nd compounds will typically be used in therapy for human subjects or patients. However, they may also be used to treat similar or identical diseases in other vertebrates such as other primates, sports animals, and pets such as horses, dogs and cats.
[0387] Suitable dosage forms, in part, depend upon the use or the route of administration, for example, oral, transdermal, transmucosal, inhalant, or by injection (parenteral). Such dosage forms should allow the compound to reach target cells. Other factors are well known in the art, and include considerations such as toxicity and dosage forms that retard the compound or composition from exerting its effects. Techniques and formulations generally may be found in Remington: The Science and Practice of Pharmacy, 21st edition, Lippincott, Williams and Wilkins, Philadelphia, PA, 2005 (hereby incorporated by reference herein).

[0388] Compounds can be formulated as pharmaceutically acceptable salts.
Pharmaceutically acceptable salts are non-toxic salts in the amounts and concentrations at which they are administered. The preparation of such salts can facilitate the pharmacological use by altering the physical characteristics of a compound without preventing it from exerting its physiological effect. Useful alterations in physical properties include lowering the melting point to facilitate transmucosal administration and increasing the solubility to facilitate administering higher concentrations of the drug.

[0389] Pharmaceutically acceptable salts include acid addition salts such as those containing sulfate, chloride, hydrochloride, fumarate, maleate, phosphate, sulfamate, acetate, citrate, lactate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluene-sulfonate, cyclohexylsulfamate and quinate. Pharmaceutically acceptable salts can be obtained from acids such as hydrochloric acid, maleic acid, sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic acid, fumaric acid, and quinic acid.

[0390] Pharmaceutically acceptable salts also include basic addition salts such as those containing benzathine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, ammonium, alkylamine, and zinc, when acidic functional groups, such as carboxylic acid or phenol are present. For example, see Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Co., Easton, PA, Vol. 2, p. 1457, 1995. Such salts can be prepared using the appropriate corresponding bases.

[0391] Pharmaceutically acceptable salts can be prepared by standard techniques. For example, the free-base form of a compound is dissolved in a suitable solvent, such as an aqueous or aqueous-alcohol in solution containing the appropriate acid and then isolated by evaporating the solution. In another exainple, a salt is prepared by reacting the free base and acid in an organic solvent.

[0392] The pharmaceutically acceptable salt of the different compounds may be present as a complex. Examples of complexes include 8-chlorotheophylline complex (analogous to, e.g., dimenhydrinate: diphenhydramine 8-chlorotheophylline (1:1) complex; Dramamine) and various cyclodextrin inclusion complexes.

[0393] Carriers or excipients can be used to produce compositions. The carriers or excipients can be chosen to facilitate administration of the compound. Examples of carriers include calcium carbonate, calcium phosphate, various sugars such as lactose, glucose, or sucrose, or types of starch, cellulose derivatives, gelatin, vegetable oils, polyethylene glycols and physiologically compatible solvents. Examples of physiologically compatible solvents include sterile solutions of water for injection (WFI), saline solution, and dextrose.

[0394] The compounds can be administered by different routes including intravenous, intraperitoneal, subcutaneous, intramuscular, oral, transmucosal, rectal, inhalant or transdermal. For oral administration, for example, the compounds can be formulated into conventional oral dosage forms such as capsules, tablets, and liquid preparations such as syrups, elixirs, and concentrated drops.

[0395] Pharmaceutical preparations for oral use can be obtained, for example, by combining the active compounds with solid excipients, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodiuin carboxymethylcellulose (CMC), and/or polyvinylpyrrolidone (PVP:
povidone). If desired, disintegrating agents may be added, such as the cross-linked polyvinylpyrrolidone, agar, or alginic acid, or a salt thereof such as sodium alginate.

[0396] Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain, for example, gum arabic, talc, poly-vinylpyrrolidone, carbopol gel, polyethylene glycol (PEG), and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dye-stuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.

[0397] Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin ("gelcaps"), as well as soft, sealed capsules made of gelatin, and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols (PEGs). In addition, stabilizers may be added.

[0398] Alternatively, injection (parenteral administration) may be used, e.g., intramuscular, intravenous, intraperitoneal, and/orsubcutaneous. For injection, the compounds of the invention are formulated in sterile liquid solutions, preferably in physiologically compatible buffers or solutions, such as saline solution, Hank's solution, or Ringer's solution. In addition, the compounds may be formulated in solid form and redissolved or suspended immediately prior to use. Lyophilized forms can also be produced.

[0399] Administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for tra.nsmucosal administration, bile salts and fusidic acid derivatives. In addition, detergents may be used to facilitate permeation. Transmucosal administration, for example, may be through nasal sprays or suppositories (rectal or vaginal).

[0400] For inhalants, compounds of the invention may be formulated as dry powder or a suitable solution, suspension, or aerosol. Powders and solutions may be fonnulated with suitable additives known in the art. For example, powders may include a suitable powder base such as lacatose or starch, and solutions may comprise propylene glycol, sterile water, ethanol, sodium chloride and other additives, such as acid, alkali and buffer salts.
Such solutions or suspensions may be administered by inhaling via spray, pump, atomizer, or nebulizer and the like. The compounds of the invention may also be used in combination with other inhaled therapies, for example corticosteroids such as fluticasone proprionate, beclomethasone dipropionate, triamcinolone acetonide, budesonide, and mometasone furoate;
beta agonists such as albuterol, salmeterol, and formoterol; anticholinergic agents such as ipratroprium bromide or tiotropium; vasodilators such as treprostinal and iloprost; enzymes such as DNAase; therapeutic proteins; immunoglobulin antibodies; an oligonucleotide, such as single or double stranded DNA or RNA, siRNA; antibiotics such as tobramycin;
muscarinic receptor antagonists; leukotriene antagonists; cytokine antagonists; protease inhibitors; cromolyn sodium; nedocril sodium; and sodium cromoglycate.

[0401] It is understood that use in combination for any route of administration includes delivery of compounds of the invention and one or more other therapeutics delivered by the same route of administration together in any formulation, including formulations where the two compounds are chemically linked such that they maintain their therapeutic activity when administered. Combination use includes administration of co-formulations or formulations of chemically joined compounds, or co-administration of the coinpounds in separate formulations.
Separate formulations may be co-administered by delivery via one device, for example the same inhalant device, the same syringe, etc., or can be co-administered from separate devices, where co-administration in this case means administered within a short time of each other.
Co-formulations of a compound of the invention and one or more additional therapies delivered by the same route includes preparation of the materials together such that they can be administered by one device, including the separate compounds combined in one formulation, or compounds that are modified such that they are chemically joined, yet still maintain their biological activity. Such chemically joined compounds may have a linkage that is substantially maintained in vivo, or the linkage may break down in vivo, separating the two active components.

[0402] The amounts of various compound to be administered can be determined by standard procedures taking into account factors such as the compound IC50, the biological half-life of the compound, the age, size, and weight of the subject, and the disorder associated with the subject. The importance of these and other factors are well known to those of ordinary skill in the art. Generally, a dose will be between about 0.01 and 50 mg/kg, preferably 0.1 and 20 mg/kg of the subject being treated. Multiple doses inay be used.

XV. Manipulation of PDE4B

[0403] As the full-length coding sequence and amino acid sequence of PDE4B
from various marrunals including human is known, cloning, construction of recombinant PDE4B, production and purification of recombinant protein, introduction of PDE4B into other organisms, and other molecular biological manipulations of PDE4B are readily performed.

[0404] Techniques for the manipulation of nucleic acids, such as, e.g., subcloning, labeling probes (e.g., random-primer labeling using Klenow polymerase, nick translation, amplification), sequencing, hybridization and the like are well disclosed in the scientific and patent literature, see, e.g., Sambrook, ed., Molecular Cloning: a Laboratory Manual (2nd ed.), Vols. 1-3, Cold Spring Harbor Laboratory, (1989); Current Protocols in Molecular Biology, Ausubel, ed. John Wiley & Sons, Inc., New York (1997); Laboratory Techniques in Biochemistry and Molecular Biology: Hybridization With Nucleic Acid Probes, Part I. Theory and Nucleic Acid Preparation, Tijssen, ed. Elsevier, N.Y. (1993).

[0405] Nucleic acid sequences can be amplified as necessary for further use using amplification methods, such as PCR, isothermal methods, rolling circle methods, etc., are well known to the skilled artisan. See, e.g., Saiki, "Amplification of Genomic DNA"
in PCR

Protocols, Innis et al., Eds., Academic Press, San Diego, CA 1990, pp 13-20;
Wharam et al., Nucleic Acids Res. 2001 Jun 1;29(11):E54-E54; Hafner et al., Biotechniques Apr;30(4):852-6, 858, 860 passim; Zhong et al., Biotechniques 2001 Apr;30(4):852-6, 858, 860 passim.

[0406] Nucleic acids, vectors, capsids, polypeptides, and the like can be analyzed and quantified by any of a number of general means well known to those of skill in the art. These include, e.g., analytical biochemical methods sucll as NMR, spectrophotometry, radiography, electrophoresis, capillary electrophoresis, high perfomiance liquid chromatography (HPLC), thin layer chromatography (TLC), and hyperdiffusion chromatography, various immunological methods, e.g. fluid or gel precipitin reactions, immunodiffusion, immuno-electrophoresis, radioimmunoassays (RIAs), enzyme-linked immunosorbent assays (ELISAs), immuno-fluorescent assays, Southern analysis, Northern analysis, dot-blot analysis, gel electrophoresis (e.g., SDS-PAGE), nucleic acid or target or signal amplification methods, radiolabeling, scintillation counting, and affinity chromatography.

[0407] Obtaining and manipulating nucleic acids used to practice the methods of the invention can be performed by cloning from genomic samples, and, if desired, screening and re-cloning inserts isolated or amplified from, e.g., genomic clones or cDNA
clones. Sources of nucleic acid used in the methods of the invention include genomic or cDNA
libraries contained in, e.g., mammalian artificial chromosomes (MACs), see, e.g., U.S. Patent Nos.
5,721,118;
6,025,155; human artificial chromosomes, see, e.g., Rosenfeld (1997) Nat.
Genet. 15:333-335;
yeast artificial chromosomes (YAC); bacterial artificial chromosomes (BAC); Pl artificial chromosomes, see, e.g., Woon (1998) Genomics 50:306-316; Pl-derived vectors (PACs), see, e.g., Kern (1997) Biotechniques 23:120-124; cosmids, recombinant viruses, phages or plasmids.

[0408] The nucleic acids of the invention can be operatively linked to a promoter. A
promoter can be one motif or an array of nucleic acid control sequences which direct transcription of a nucleic acid. A promoter can include necessary nucleic acid sequences near the start site of transcription, such as, in the case of a polymerase II type promoter, a TATA
element. A promoter also optionally includes distal enhancer or repressor elements which can be located as much as several thousand base pairs from the start site of transcription. A
"constitutive" promoter is a promoter which is active under most environmental and developmental conditions. An "inducible" promoter is a promoter wlzich is under environmental or developmental regulation. A "tissue specific" promoter is active in certain tissue types of an organism, but not in other tissue types from the same organism. The term "operably linked" refers to a functional linkage between a nucleic acid expression control sequence (such as a promoter, or array of transcription factor binding sites) and a second nucleic acid sequence, wherein the expression control sequence directs transcription of the nucleic acid corresponding to the second sequence.

[0409] The nucleic acids of the invention can also be provided in expression vectors and cloning vehicles, e.g., sequences encoding the polypeptides of the invention.
Expression vectors and cloning vehicles of the invention can comprise viral particles, baculovirus, phage, plasmids, phagemids, cosmids, fosmids, bacterial artificial chromosomes, viral DNA (e.g., vaccinia, adenovirus, foul pox virus, pseudorabies and derivatives of SV40), Pl-based artificial chromosomes, yeast plasmids, yeast artificial chromosomes, and any other vectors specific for specific hosts of interest (such as bacillus, Aspergillus and yeast). Vectors of the invention can include chromosomal, non-chromosomal and synthetic DNA sequences. Large numbers of suitable vectors are known to those of skill in the art, and are commercially available.

[0410] The nucleic acids of the invention can be cloned, if desired, into any of a variety of vectors using routine molecular biological methods; methods for cloning in vitro amplified nucleic acids are disclosed, e.g., U.S. Pat. No. 5,426,039. To facilitate cloning of amplified sequences, restriction enzyme sites can be "built into" a PCR primer pair.
Vectors may be introduced into a genome or into the cytoplasm or a nucleus of a cell and expressed by a variety of conventional techniques, well described in the scientific and patent literature. See, e.g., Roberts (1987) Nature 328:731; Schneider (1995) Protein Expr. Purif.
6435:10;
Sambrook, Tijssen or Ausubel. The vectors can be isolated from natural sources, obtained from such sources as ATCC or GenBank libraries, or prepared by synthetic or recombinant methods. For example, the nucleic acids of the invention can be expressed in expression cassettes, vectors or viruses which are stably or transiently expressed in cells (e.g., episomal expression systems). Selection markers can be incorporated into expression cassettes and vectors to confer a selectable phenotype on transformed cells and sequences.
For example, selection markers can code for episomal maintenance and replication such that integration into the host genome is not required.

[0411] The nucleic acids can be administered in vivo for in situ expression of the peptides or polypeptides of the invention. The nucleic acids can be administered as "naked DNA" (see, e.g., U.S. Patent No. 5,580,859) or in the form of an expression vector, e.g., a recombinant virus. The nucleic acids can be administered by any route, including peri- or intra-tumorally, as described below. Vectors administered in vivo can be derived from viral genomes, including recombinantly modified enveloped or non-enveloped DNA and RNA
viruses, preferably selected from baculoviridiae, parvoviridiae, picomoviridiae, herpesveridiae, poxviridae, adenoviridiae, or picornnaviridiae. Cliimeric vectors may also be employed which exploit advantageous merits of each of the parent vector properties (See e.g., Feng (1997) Nature Biotechnology 15:866-870). Such viral genomes may be modified by recombinant DNA techniques to include the nucleic acids of the invention; and may be further engineered to be replication deficient, conditionally replicating or replication competent.
In alternative aspects, vectors are derived from the adenoviral (e.g., replication incompetent vectors derived from the human adenovirus genome, see, e.g., U.S. Patent Nos. 6,096,718;
6,110,458;
6,113,913; 5,631,236); adeno-associated viral and retroviral genomes.
Retroviral vectors can include those based upon murine leukemia virus (MuLV), gibbon ape leukemia virus (GaLV), Simian Immuno deficiency virus (SIV), human immuno deficiency virus (HIV), and combinations thereof; see, e.g., U.S. Patent Nos. 6,117,681; 6,107,478;
5,658,775; 5,449,614;
Buchscher (1992) J. Virol. 66:2731-2739; Johann (1992) J. Virol. 66:1635-1640). Adeno-associated virus (AAV)-based vectors can be used to transduce cells with target nucleic acids, e.g., in the in vitro production of nucleic acids and peptides, and in in vivo and ex vivo gene therapy procedures; see, e.g., U.S. Patent Nos. 6,110,456; 5,474,935; Okada (1996) Gene TZzeY.
3:957-964.

[0412] The present invention also relates to fusion proteins, and nucleic acids encoding them.
A polypeptide of the invention can be fused to a heterologous peptide or polypeptide, such as N-terminal identification peptides which impart desired characteristics, such as increased stability or simplified purification. Peptides and polypeptides of the invention can also be synthesized and expressed as fusion proteins with one or more additional domains linked tllereto for, e.g., producing a more immunogenic peptide, to more readily isolate a recombinantly synthesized peptide, to identify and isolate antibodies and antibody-expressing B cells, and the like. Detection and purification facilitating domains include, e.g., metal chelating peptides such as polyhistidine tracts and histidine-tryptophan modules that allow purification on immobilized metals, protein A domains that allow purification on immobilized immunoglobulin, and the domain utilized in the FLAGS extension/affinity purification system (Immunex Corp, Seattle WA). The inclusion of a cleavable linker sequences such as Factor Xa or enterokinase (Invitrogen, San Diego CA) between a purification domain and the motif-comprising peptide or polypeptide to facilitate purification. For example, an expression vector can include an epitope-encoding nucleic acid sequence linked to six histidine residues followed by a thioredoxin and an enterokinase cleavage site (see e.g., Williams (1995) Biochemistry 34:1787-1797; Dobeli (1998) Protein Expr. Purif. 12:404-414). The histidine residues facilitate detection and purification wliile the enterokinase cleavage site provides a means for purifying the epitope from the remainder of the fusion protein. In one aspect, a nucleic acid encoding a polypeptide of the invention is asseinbled in appropriate phase with a leader sequence capable of directing secretion of the translated polypeptide or fragment thereof. Technology pertaining to vectors encoding fusion proteins and application of fusion proteins are well disclosed in the scientific and patent literature, see e.g., Kroll (1993) DNA
Cell. Biol. 12:441-53.

[0413] The nucleic acids and polypeptides of the invention can be bound to a solid support, e.g., for use in screening and diagnostic methods. Solid supports can include, e.g., membranes (e.g., nitrocellulose or nylon), a microtiter dish (e.g., PVC, polypropylene, or polystyrene), a test tube (glass or plastic), a dip stick (e.g., glass, PVC, polypropylene, polystyrene, latex and the like), a microfiige tube, or a glass, silica, plastic, metallic or polymer bead or other substrate such as paper. One solid support uses a metal (e.g., cobalt or nickel)-comprising column which binds with specificity to a histidine tag engineered onto a peptide.

[0414] Adhesion of molecules to a solid support can be direct (i.e., the molecule contacts the solid support) or indirect (a "linker" is bound to the support and the molecule of interest binds to this linker). Molecules can be immobilized either covalently (e.g., utilizing single reactive thiol groups of cysteine residues (see, e.g., Colliuod (1993) Bioconjugate Chem. 4:528-536) or non-covalently but specifically (e.g., via immobilized antibodies (see, e.g., Schuhmann (1991) Adv. Mater. 3:388-391; Lu (1995) Anal. Chem. 67:83-87; the biotin/strepavidin system (see, e.g., Iwane (1997) Bioplzys. Biochern. Res. Comm. 230:76-80); metal chelating, e.g., Langmuir-Blodgett films (see, e.g., Ng (1995) Langinuir 11:4048-55); metal-chelating self-assembled monolayers (see, e.g., Sigal (1996) Anal. Cliem. 68:490-497) for binding of polyhistidine fusions.

[0415] Indirect binding can be achieved using a variety of linkers which are commercially available. The reactive ends can be any of a variety of functionalities including, but not limited to: amino reacting ends such as N-hydroxysuccinimide (NHS) active esters, imidoesters, aldehydes, epoxides, sulfonyl halides, isocyanate, isothiocyanate, and nitroaryl halides; and thiol reacting ends such as pyridyl disulfides, maleimides, thiophthalimides, and active halogens. The heterobifunctional crosslinking reagents have two different reactive ends, e.g., an amino-reactive end and a thiol-reactive end, while homobifitnctional reagents have two similar reactive ends, e.g., bismaleimidohexane (BMH) which permits the cross-linking of sulfhydryl-containing compounds. The spacer can be of varying length and be aliphatic or aromatic. Examples of commercially available homobifunctional cross-linking reagents include, but are not limited to, the imidoesters such as dimethyl adipimidate dihydrochloride (DMA); dimethyl pimelimidate dihydrochloride (DMP); and dimethyl suberimidate dihydrochloride (DMS). Heterobifunctional reagents include commercially available active halogen-NHS active esters coupling agents such as N-succinimidyl bromoacetate and N-succinimidyl (4-iodoacetyl)aminobenzoate (SIAB) and the sulfosuccinimidyl derivatives such as sulfosuccinimidyl(4-iodoacetyl)aminobenzoate (sulfo-SIAB) (Pierce). Another group of coupling agents is the heterobifunctional and thiol cleavable agents such as N-succinimidyl 3-(2-pyridyidithio)propionate (SPDP) (Pierce Chemicals, Rockford, IL).

[0416] Antibodies can also be used for binding polypeptides and peptides of the invention to a solid support. This can be done directly by binding peptide-specific antibodies to the colunm or it can be done by creating fusion protein chimeras comprising motif-containing peptides linked to, e.g., a known epitope (e.g., a tag (e.g., FLAG, myc) or an appropriate immunoglobulin constant domain sequence (an "immunoadhesin," see, e.g., Capon (1989) Nature 377:525-531 (1989).

[0417] Nucleic acids or polypeptides of the invention can be immobilized to or applied to an array. Arrays can be used to screen for or monitor libraries of compositions (e.g., small molecules, antibodies, nucleic acids, etc.) for their ability to bind to or modulate the activity of a nucleic acid or a polypeptide of the invention. For example, in one aspect of the invention, a monitored parameter is transcript expression of a gene comprising a nucleic acid of the invention. One or more, or, all the transcripts of a cell can be measured by hybridization of a sample comprising transcripts of the cell, or, nucleic acids representative of or complementary to transcripts of a cell, by hybridization to immobilized nucleic acids on an array, or "biochip."

By using an "array" of nucleic acids on a microchip, some or all of the transcripts of a cell can be simultaneously quantified. Alternatively, arrays comprising genomic nucleic acid can also be used to determine the genotype of a newly engineered strain made by the methods of the invention. Polypeptide arrays" can also be used to simultaneously quantify a plurality of proteins.

[0418] The terms "array" or "microarray" or "biochip" or "chip" as used herein is a plurality of target elements, each target element comprising a defined amount of one or more polypeptides (including antibodies) or nucleic acids imtnobilized onto a defmed area of a substrate surface. In practicing the methods of the invention, any known array and/or method of making and using arrays can be incorporated in whole or in part, or variations thereof, as disclosed, for example, in U.S. Patent Nos. 6,277,628; 6,277,489; 6,261,776;
6,258,606;
6,054,270; 6,048,695; 6,045,996; 6,022,963; 6,013,440; 5,965,452; 5,959,098;
5,856,174;
5,830,645; 5,770,456; 5,632,957; 5,556,752; 5,143,854; 5,807,522; 5,800,992;
5,744,305;
5,700,637; 5,556,752; 5,434,049; see also, e.g., WO 99/51773; WO 99/09217; WO
97/46313;
WO 96/17958; see also, e.g., Johnston (1998) Curr. Biol. 8:R171-R174;
Schuminer (1997) Biotechyaiques 23:1087-1092; Kern (1997) Biotechniques 23:120-124; Solinas-Toldo (1997) Genes, Chromosonzes & Cancef 20:399-407; Bowtell (1999) Nature Genetics Supp.
21:25-32.
See also published U.S. patent applications Nos. 20010018642; 20010019827;
20010016322;
20010014449; 20010014448; 20010012537; 20010008765.

Host Cells and Transformed Cells [0419] The invention also provides a transformed cell comprising a nucleic acid sequence of the invention, e.g., a sequence encoding a polypeptide of the invention, or a vector of the invention. The host cell may be any of the host cells familiar to those skilled in the art, including prokaryotic cells, eukaryotic cells, such as bacterial cells, fungal cells, yeast cells, mammalian cells, insect cells, or plant cells. Exemplary bacterial cells include E. coli, Streptomyces, Bacillus subtilis, Salmonella typhirnurium and various species within the genera Pseudonaon.as, Streptomyces, and Staphylococcus. Exemplary insect cells include Drosophila S2 and Spodoptera Sf9. Exemplary animal cells include CHO, COS or Bowes melanoma or any mouse or human cell line. The selection of an appropriate host is within the abilities of those skilled in the art.

[0420] Vectors may be introduced into the host cells using any of a variety of techniques, including transformation, transfection, transduction, viral infection, gene guns, or Ti-mediated gene transfer. Particular methods include calcium phosphate transfection, DEAE-Dextran mediated transfection, lipofection, or electroporation.

[0421] Engineered host cells can be cultured in conventional nutrient media modified as appropriate for activating promoters, selecting transformants or amplifying the genes of the invention. Following transformation of a suitable host strain and growth of the host strain to an appropriate cell density, the selected promoter may be induced by appropriate means (e.g., temperature shift or chemical induction) and the cells may be cultured for an additional period to allow them to produce the desired polypeptide or fragment thereof.

[0422] Cells can be harvested by centrifugation, disrupted by physical or chemical means, and the resulting crude extract is retained for further purification.
Microbial cells employed for expression of proteins can be disrupted by any convenient method, including freeze-thaw cycling, sonication, mechanical disruption, or use of cell lysing agents. Such methods are well known to those skilled in the art. The expressed polypeptide or fragment can be recovered and purified from recombinant cell cultures by methods including ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose clzromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography. Protein refolding steps can be used, as necessary, in completing configuration of the polypeptide. If desired, high performance liquid chromatography (HPLC) can be employed for final purification steps.
[0423] Various mammalian cell culture systems can also be employed to express recombinant protein. Examples of mammalian expression systeins include the COS-7 lines of monkey kidney fibroblasts and other cell lines capable of expressing proteins from a compatible vector, such as the C127, 3T3, CHO, HeLa and BHK cell lines.

[0424] The constructs in host cells can be used in a conventional manner to produce the gene product encoded by the recombinant sequence. Depending upon the host employed in a recombinant production procedure, the polypeptides produced by host cells containing the vector may be glycosylated or may be non-glycosylated. Polypeptides of the invention may or may not also include an initial methionine amino acid residue.

[0425] Cell-free translation systems can also be employed to produce a polypeptide of the invention. Cell-free translation systems can use mRNAs transcribed from a DNA
construct comprising a promoter operably linked to a nucleic acid encoding the polypeptide or fragment thereof. In some aspects, the DNA construct may be linearized prior to conducting an in vitro transcription reaction. The transcribed mRNA is then incubated with an appropriate cell-free translation extract, such as a rabbit reticulocyte extract, to produce the desired polypeptide or fragment thereof.

[0426] The expression vectors can contain one or more selectable marker genes to provide a phenotypic trait for selection of transformed host cells such as dihydrofolate reductase or neomycin resistance for eukaryotic cell culture, or such as tetracycline or ampicillin resistance in E. coli.

[0427] For transient expression in mammalian cells, cDNA encoding a polypeptide of interest may be incorporated into a mammalian expression vector, e.g. pcDNA1, which is available commercially from Invitrogen Corporation (San Diego, Calif., U.S.A.;
catalogue number V490-20). This is a multifunctional 4.2 kb plasmid vector designed for cDNA
expression in eukaryotic systems, and cDNA analysis in prokaryotes, incorporated on the vector are the CMV promoter and enhancer, splice segment and polyadenylation signal, an SV40 and Polyoma virus origin of replication, and M13 origin to rescue single strand DNA for sequencing and mutagenesis, Sp6 and T7 RNA promoters for the production of sense and anti-sense RNA transcripts and a Col El-like higlz copy plasmid origin. A
polylinker is located appropriately downstream of the CMV promoter (and 3' of the T7 promoter).

[0428] The cDNA insert may be first released from the above phagemid incorporated at appropriate restriction sites in the pcDNAI polylinker. Sequencing across the junctions may be performed to confirm proper insert orientation in pcDNAI. The resulting plasmid may then be introduced for transient expression into a selected mammalian cell host, for example, the monkey-derived, fibroblast like cells of the COS-1 lineage (available from the American Type Culture Collection, Rockville, Md. as ATCC CRL 1650).

[0429] For transient expression of the protein-encoding DNA, for example, COS-1 cells may be transfected with approximately 8 g DNA per 106 COS cells, by DEAE-mediated DNA
transfection and treated with chloroquine according to the procedures described by Sambrook et al, Molecular Cloning: A Laboratory Manual, 1989, Cold Spring Harbor Laboratory Press, Cold Spring Harbor N.Y, pp. 16.30-16.37. An exemplary method is as follows.
Briefly, COS-1 cells are plated at a density of 5 x 106 cells/dish and then grown for 24 hours in FBS-supplemented DMEM/F12 medium. Medium is then removed and cells are washed in PBS
and then in medium. A transfection solution containing DEAE dextran (0.4 mg/ml), 100 M
chloroquine, 10% NuSerum, DNA (0.4 mg/ml) in DMEM/F12 medium is then applied on the cells 10 ml volume. After incubation for 3 hours at 37 C, cells are washed in PBS and medium as just described and then shocked for 1 minute with 10% DMSO in mediuin. Cells are allowed to grow for 2-3 days in 10% FBS-supplemented medium, and at the end of incubation dishes are placed on ice, washed witli ice cold PBS and then removed by scraping. Cells are then harvested by centrifugation at 1000 rpm for 10 minutes and the cellular pellet is frozen in liquid nitrogen, for subsequent use in protein expression. Northern blot analysis of a thawed aliquot of frozen cells may be used to confirm expression of receptor-encoding cDNA in cells under storage.

[0430] In a like manner, stably transfected cell lines can also prepared, for example, using two different cell types as host: CHO K1 and CHO Pro5. To construct these cell lines, cDNA
coding for the relevant protein may be incorporated into the mammalian expression vector pRC/CMV (Invitrogen), which enables stable expression. Insertion at this site places the cDNA under the expression control of the cytomegalovirus promoter and upstream of the polyadenylation site and terminator of the bovine growth hormone gene, and into a vector background comprising the neomycin resistance gene (driven by the SV40 early promoter) as selectable marker.

[0431] An exemplary protocol to introduce plasmids constructed as described above is as follows. The host CHO cells are first seeded at a density of 5x105 in 10% FBS-supplemented MEM medium. After growth for 24 hours, fresh medium is added to the plates and three hours later, the cells are transfected using the calcium phosphate-DNA co-precipitation procedure (Sambrook et al, supra). Briefly, 3 g of DNA is mixed and incubated with buffered calcium solution for 10 minutes at room temperature. An equal volume of buffered phosphate solution is added and the suspension is incubated for 15 minutes at room temperature.
Next, the incubated suspension is applied to the cells for 4 hours, removed and cells were shocked with medium containing 15% glycerol. Three minutes later, cells are washed with medium and incubated for 24 hours at normal growth conditions. Cells resistant to neomycin are selected in 10% FBS-supplemented alpha-MEM medium containing G418 (1 mg/ml). Individual colonies of G418-resistant cells are isolated about 2-3 weeks later, clonally selected and then propagated for assay purposes.

EXAMPLES
[0432] A number of examples involved in the present invention are described below. In most cases, alternative techniques could also be used. The examples are intended to be illustrative and are not limiting or restrictive to the scope of the invention.

Example 1: Synthesis of compounds of Formula Ia Scheme -1 v%W-' v -~W" v -w' x x 6tN y Step 1 1Step 2Br y Step3 )No H t~N t N

=
m Prot iv Prot II =

v%~\ v7::~W\ v--~W\
x A \ y 11 Step 4 A ~ y Step 5~ A \ y 11 t~N t NH t~N
=
Prot = k V VI
Formula Ia Step -1 Synthesis of Fornzula III
[0433] Compound of Formula ITI can be prepared from commercially available compound II
by reacting with a reagent containing a leaving group, e.g. chloro, tosyl, etc., in presence of a base, e.g. triethyl amine, pyridine, aqueous hydroxides, etc., in a polar solvent, e.g, dimethylformainide (DMF), or water (Greene, T. W.; Wuts, P.G.M. Protective Groups in Organic S'ynthesis, 3rd ed.; John Wiley & Sons: New York, 1999). The product can be isolated by conventional work up procedure, e.g. extraction of the product with an organic solvent and evaporation.

Step -2 Synthesis of Formula IV
[0434] Compound of Formula IV can be prepared from compound III by reacting with bromine in carbon tetrachloride or N-bromosuccinimide in THF. The product can be isolated by conventional work up procedure, e.g. extraction of the product with an organic solvent and purifying by column chromatography.

Step -3 Synthesis of Formula V
[0435] Compound of Formula V can be prepared from compound IV by reacting with boronic acids under Suzuki reaction conditions (Smith M.B.; March, J. March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5'lZ ed.; John Wiley & Sons: New York, 2001 p 868). Alternately, compound IV can also be reacted with tin, zinc, or copper reagents, under Stille, Negislii or cuprate coupling reaction conditions, respectively (Smith M.B.; March, J. March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5'ji ed.; John Wiley & Sons: New York, 2001 p 931), to provide compound V. The product can be isolated by conventional work up procedure, e.g. extraction of the product with an organic solvent and purifying by column chromatography.

Step -4 Synthesis of foYmula VI
[0436] Compound of formula VI can be prepared by reacting compound V with fluorides (for silyl protecting groups - tetrabutyl ammonium fluoride or ammonium fluoride) (Greene, T. W.; Wuts, P.G.M. Protective Groups in Organic Synthesis, 3d ed.; John Wiley & Sons:
New York, 1999, p 620), base (for aryl sulfone protecting groups - aqueous potassium hydroxide) (Greene, T. W.; Wuts, P.G.M. Protective Groups in Organic Syntliesis, 3rd ed.;
John Wiley & Sons: New York, 1999, p 615), or an acid (for carbamates -trifluoroacetic acid) (Greene, T. W.; Wuts, P.G.M. Protective Groups in Orgaiaic Synthesis, 3rd ed.;
John Wiley &
Sons: New York, 1999, p 272) a.nd isolating the product following standard workup procedures, e.g. extraction of the product with organic solvent and purifying by column chromatography.

Step -5 Synthesis of Formula Ia [0437] Compounds of Formula Ia can be prepared by reacting compound VI with nucleophilic reagents, e.g. acid chlorides, sulfonyl chlorides, isocyanates, isothiocyanates, alkyl halides, benzyl halides, etc., under basic conditions (General reference: Smith M.B.;
March, J. March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5'j' ed.; John Wiley & Sons: New York, 2001). The product can be isolated by following standard workup procedures, e.g. extraction of the product with organic solvent and purifying by colurnn chromatography.

Example 2: Alternate synthesis of compounds of Formula Ia Scheme - 2 vz:~W~'vGW\v~ =
x x x Y Step 1Br y Step 2410 Br~Y Step 3 t-NH t-NH t N
=
vu iv Prot II

v%W~ vl::W\
y Stepd~ A 1~ y Ste p5 ' A y t~N = t'~NH t~N
Prot k V vi Formula Ia Step -1 Synthesis of Formula VII
[0438] Compound of Formula VII can be prepared from commercially available compound II by reacting with bromine in carbon tetrachloride at low temperature, for example -20 C, or N-bromosuccinimide in THF. The product can be isolated by conventional work up procedure, e.g. extraction of the product with an organic solvent and purifying by column chromatography.
Step -2 Syntlzesis of Fortnula IV
[0439] Compound of Formula IV can be prepared from compound VII by reacting with a reagent containing a leaving group, e.g. chloro, tosyl, etc., in presence of a base, e.g. triethyl amine, pyridine, aqueous hydroxides, etc., in a polar solvent, e.g, dimethylformamide (DMF), or water. The product can be isolated by conventional work up procedure, e.g.
extraction of the product with an organic solvent and evaporation.

Step -3 Synthesis of Foymula V
[0440] Compound of Formula V can be prepared from compound IV by reacting with boronic acids under Suzuki reaction conditions. Alternately, compound IV can also be reacted with tin, zinc, or copper reagents, under Stille, Negishi or cuprate coupling reaction conditions respectively, to provide compound V. The product can be isolated by conventional work up procedure, e.g. extraction of the product with an organic solvent and purifying by column chromatography.

Step -4 Synthesis offormula VI
[0441] Compound of Fonnula VI can be prepared by reacting compound V with fluorides (for silyl protecting groups - tetrabutyl ammonium fluoride, or ammonium fluoride), base (for aryl sulfone protecting groups - aqueous potassium hydroxide), or an acid (for carbamates -trifluoroacetic acid) and isolating the product following standard workup procedures, e.g.
extraction of the product with organic solvent and purifying by column chromatography.

Step -5 Synthesis of FoYmula Ia [0442] Compound of Forinula Ia can be prepared by reacting compound VI with nucleopliilic reagents, e.g. acid chlorides, sulfonyl chlorides, isocyanates, isothiocyanates, etc., under basic conditions. The product can be isolated by following standard workup procedures, e.g. extraction of the product with organic solvent and purifying by coluinn chromatography.
Example 3: Alternate synthesis of compounds of Formula Ia Scheme - 3 V%W-' x V ~W\
Br y Step 1(R120)26 , ~ y Step y ~
t--N t--N t--N
IV x Prot Prot Prot VIII V

v%~\ X
Step 3 A',\ y/ Ste A 1\

t-NH t-~N
k vi Formula Ia Step -1 Synthesis of Fornaula VIII
[0443] Compound of Formula VIII can be prepared from compound IV by reacting with the tributyl ester of boronic acid as described by Gilman et al. in J. Am. Chem.
Soc., 1957, 79, 3077. The product can be isolated by conventional work up procedure, e.g.
extraction of the product with an organic solvent and purifying by column chromatography.

Step -2 Synthesis of Formula V
[0444] Compound of Formula V can be prepared by reacting compound VIII with compounds of Formula A-Br or A-I under Suzuki reaction conditions and isolating the product following standard workup procedures, e.g. extraction of the product with organic solvent and purifying by column chromatography.

Step -3 Synthesis of Fonmula VI
[0445] Compound of Formula VI can be prepared by reacting compound V with fluorides (for silyl protecting groups - tetrabutyl ammonium fluoride, or ammonium fluoride), base (for aryl sulfone protecting groups - aqueous potassium hydroxide), or an acid (for carbamates -trifluoroacetic acid) and isolating the product following standard workup procedures, e.g.
extraction of the product with organic solvent and purifying by column chromatography Step -4 Synthesis of Formula Ia [0446] Compound of Formula Ia can be prepared by reacting compound VI with nucleophilic reagents, e.g. acid chlorides, sulfonyl chlorides, isocyanates, isothiocyanates, etc., under basic conditions. The product can be isolated by following standard workup procedures, e.g. extraction of the product with organic solvent and purifying by column chromatography.
Example 4: Alternate synthesis of compounds of Formula Ia Scheme - 4 v~W
v:W v~W A' ~x ~
Br ~x Br ~
y Stepl & yX Step2 t N y t.N t.N =k *k Formula Ia VII
IX
Step -1 Synthesis of Fornzula IX
[0447] Compound of Formula IX can be prepared by reacting compound VII with nucleophilic reagents, e.g. acid chlorides, sulfonyl chlorides, isocyanates, isothiocyanates, alkyl halides, benzyl halides, etc., under basic conditions. The product can be isolated by following standard workup procedures, e.g. extraction of the product with organic solvent and purifying by column chromatography.

Step -2 Synthesis of Formula Ia [0448] Compound of Formula Ia can be prepared from coinpound IX by reacting with boronic acids under Suzuki reaction conditions (aqueous base and Pd(0) catalyst or anhydrous conditions with KF in dioxane with Pd(0) catalyst). Alternately, coinpound IX
can also be reacted with a tin reagent, zinc reagent or copper reagent, under Stille, Negishi or cuprate coupling reaction conditions respectively, to provide compound Ia. The product can be isolated by conventional work up procedure, e.g. extraction of the product with an organic solvent and purifying by column chromatography.

Example 5: Alternate synthesis of compounds of Formula Ia Scheme - 5 =W
Br ~ N'x (R120)26 ~N x A v~W x 11 y Stepl I y Step2 y t~N t~N

k k k ix X Formula Ia Step -1 Synthesis of Forrnula X
[0449] Compound of Formula X can be prepared from compound IX by reacting with tributyl ester of boronic acid as described by Gilman et. al. in J. Am. Chem.
Soc., 1957, 79, 3077. The product can be isolated by conventional work up procedure, e.g.
extraction of the product with an organic solvent and purifying by column chromatography.

Step -2 Synthesis of Formula Ia [0450] Compound of Formula Ia can be prepared by reacting compound X with compounds of Formula A-Br or A-I under Suzuki reaction conditions and isolating the product following standard workup procedures, e.g. extraction of the product with organic solvent and purifying by column chromatography.

[0451] Example 6: Alternate synthesis of compounds of Formula Ia Scheme - 6 Br v-z-W% v~W v'W
' ~- e x ~ Stepl A ~ X Step2 Y 01- Y t Y
t"N tN N
H H jk viI vi Formula Ia Step -1 Synthesis of FoYnaula VI
[0452] Compound of Formula VI can be prepared from compound VII by reacting with boronic acids under Suzuki reaction conditions (aqueous base and Pd(O) catalyst or anhydrous conditions with KF in dioxane with Pd(O) catalyst). Alternately, compound VII
can also be reacted witli a tin reagent, zinc reagent or copper reagent, under Stille, Negishi or cuprate coupling reaction conditions respectively, to provide compound VI. The product can be isolated by conventional work up procedure, e.g. extraction of the product with an organic solvent and purifying by column chromatography.

Step -2 Syntlaesis of Formula Ia [0453] Compound of Formula Ia can be prepared by reacting compound VI with nucleophilic reagents, e.g. acid chlorides, sulfonyl chlorides, isocyanates, isothiocyanates, alkyl halides, benzyl halides, etc., under basic conditions. The product can be isolated by following standard workup procedures, e.g. extraction of the product with organic solvent and purifying by column chromatography.

Example 7: Synthesis of intermediate 3-(3,4-dimethoxy-phenyl)-1H-pyrrolo [2,3-b]pyridine 1 Scheme - 7 Br Br ~~~ Step 1 ~OXT Step N N
'N N H g O O
S:O

d O
O /O
11 Step 3 / N Step 4 N
N N
0 $:O H
U

Step 1 - Preparation of 3-bromo-IH-pyrrolo[2,3-bJpyridine 3 [0454] 7-Azaindole (2, 3.57 g, 30.2 mmol) was dissolved in tetrahydrofuran (240 mL) under an atmosphere of nitrogen. At -40 C, N-bromosuccinimide (5.38 g, 30.2 inmol) in tetrahydrofura.n was added under an atmosphere of nitrogen. The reaction mixture was stirred for a few hours as it was gradually warmed to room temperature and the reaction was followed by TLC. The reaction was quenched with sodium thiosulfate pentahydrate (7.50 g, 30.2 mmol) in water (1M). Two layers were separated, and the aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with water, brine, dried with anhydrous sodium sulfate, and evaporated to dryness. The crude material was purified by column chromatography (25 - 40% ethyl acetate in hexanes) to yield the desired product as a white solid, 3, (4.20 g, 21.3 mmol). MS(ESI) [M+H+]+ = 198.5.

Step 2- Preparation of I-benzenesulfonyl-3-bromo-1 H-pyrrolo[2, 3-bJpyridine 4 [0455] 3-Bromo-lH-pyrrolo[2,3-b]pyridine (3, 280 mg, 1.4 mmol) was dissolved in acetone (15 mL) and potassium carbonate (220 mg, 1.6 mmol) was added, followed by benzenesulfonyl chloride (0.2 mL, 1.6 mmol). The reaction mixture was heated to reflux overnight, filtered and concentrated under reduced pressure. The resulting solid was purified by flash chromatography (5% - 20% ethyl acetate: hexanes) to provide the desired product, 4, (300 mg, 47%). MS(ESI) [M+H+]+= 455Ø

Step 3-PNeparation of 1-benzenesulfonyl-3-(3,4-dimethoxy phenyl)-IHpyrrolo[2,3-bJpyridine 5 [0456] 1-Benzenesulfonyl-3-bromo-lH-pyrrolo[2,3-b]pyridine (4, 1.00 g, 2.96 mmol) was dissolved in tetrahydrofuran (16 mL) and 3,4-dimethoxyphenyl boronic acid (1.35 g, 7.41 mmol), tetrakis(triphenylphosphine)palladium(0) (200 mg, 0.1 mmol), and 1 M
potassium carbonate (8 mL) were added. The reaction mixture was heated in a CEM Discover microwave at 120 C for 10 minutes. The reaction mixture was concentrated under reduced pressure and partitioned between ethyl acetate and water. The organic portions were dried with anhydrous magnesium sulfate, filtered, and the filtrate was adsorbed onto silica. The mixture was purified by flash chromatograpliy (30% ethyl acetate: hexanes) to provide the desired product, 5, (909 mg, 78%). MS(ESI) [M+H+]+= 394.9.

Step 4-Preparation of 3-(3,4-dimethoxyphenyl)-1Hpyf rolo[2,3-bJpyridine 1 [0457] 1-Benzenesulfonyl-3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine (5, 290 mg, 0.74 mmol) was dissolved in ethanol (4 mL) and potassium hydroxide pellets (330 mg, 5.9 mmol) were added. The reaction was heated in a CEM Discover microwave instrument at 120 C for 10 minutes. The reaction mixture was concentrated to dryness and the residue was partitioned between ethyl acetate and saturated sodium bicarbonate. The organic portions were dried with anhydrous magnesium sulfate, filtered and the filtrate concentrated to provide the desired product, 1, which was used without further purification (191 mg).
MS(ESI) [M+H+]+=255.1.

Example 8: Alternative Synthesis of the key intermediate 3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 1 Scheme - 8 \ O

1 = O' Br Br Step Step 2 N H N ~O N H

Step-1 Preparation of 3-Bromo pyrrolo[2,3-b]pyridine-l-carboxylic acid tert-butyl ester 6 [0458] Into a round bottom flask was added 3-bromo-7-azaindole (3, 2.6 g, 13.0 mmol) and N,N-dimethylformamide (50 mL) and sodium hydride (60% dispersion in mineral oil) (550 mg, 14.0 mmol) under an atmosphere of nitrogen. The reaction mixture was stirred at room temperature for 30 minutes, followed by addition of di-tert-butyldicarbonate (4.0 g, 18.0 mmol). The reaction mixture was stirred at room temperature overnight and was poured into water and extracted into ethyl acetate. The organic layer was washed with brine and dried over anhydrous sodium sulfate. The crude material was purified by column chromatography (30%
ethyl acetate in hexane) to yield the desired product, 6, as a solid (3.0 g, 10.1 mmol).

Step 2- PYeparation of 3-(3, 4-dimethoxy phenyl)-1 H pyf-nolo[2, 3-bJpyridine [0459] Into a round bottom flask was added 3-Bromo-pyrrolo[2,3-b]pyridine-l-carboxylic acid tert-butyl ester (6, 2.8 g, 9.4 mmol) and tetrahydrofuran (100 mL).
Tetrakis(triphenylphosphine)palladium(0) (500 mg, 0.50 mmol), 3,4-dimethoxyphenylboronic acid (2.1 g, 11 mmol) and 1 M K2C03 solution (50 mL) The reaction mixture was stirred at 65 C overnight. The reaction mixture was poured into water and extracted into ethyl acetate. The organic layer was washed with brine and dried over anhydrous magnesium sulfate. The crude material was purified by column chromatography (gradient systein of hexanes and ethyl acetate) to yield the titled compound, 1, (500 mg, 2 mmol, 20%) and 3-(3,4-dimethoxy-phenyl)-pyrrolo[2,4-b]pyridine-l-carboxylic acid tert-butyl ester (2.1 g, 5.9 mmol, 63%). The later compound can be easily converted to the desired compound, 1 by treatment with acid reagents such as TFA or HCl.

Example 9: Synthesis of 8-[3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-sulfonyl]-quinoline 7 Scheme - 9 /

= _ 0 0 =/

0 = / ~ ~ ~ 1 N N
N N
H ~ O
N

Preparation of 8-[3-(3,4-dimethoxyphenyl) pyrrolo[2,3-bJpyridine-l-sulfonylJ-quinoline 7 [0460] 3-(3,4-Dimethoxy-phenyl)-1H-pyrrolo-[2,3-b]pyridine, 1, (50 mg, 0.20 mmol) was dissolved in methylene chloride (4 mL). Aqueous potassium hydroxide (50%
wt/vol, 300 gL) and tetrabutylammonium hydrogen sulfate (2 mg, 0.007 nmtnol) were added. The reaction mixture was stirred for 10 minutes at room temperature. Into the reaction was added 8-quinoline-sulfonyl chloride (48 mg, 0.21 mmol) and the reaction mixture was stirred for 2 hours at room temperature. The reaction mixture was concentrated and the residue was washed with brine and ethyl acetate. The organic portion was dried with anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by preparative TLC (75% ethyl acetate: hexanes). The product was eluted from the silica with ethyl acetate and filtration. The filtrate was concentrated to provide 7 (14 mg, 16 %). MS(ESI) [M+H+]+=445.9.

Example 10: Synthesis of 3-(3,4-dimethoxy-phenyl)-1-phenylmethanesulfonyl-lH-pyrrolo[2,3-b]pyridine 8 i o N N
6~3 [0461] 3-(3,4-Diinethoxy-phenyl)-1-phenylmethanesulfonyl-lH-pyrrolo[2,3-b]pyridine 8 was prepared using the same protocol as described in Example 9, substituting 8-quinoline-sulfonyl chloride with phenyl-methanesulfonyl chloride. MS(ESI) [M+H+]+=409Ø
Example 11: Synthesis of 1-(3-chloro-phenylmethanesulfonyl)-3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 9 /
O

O
=~
N N

~O

CI

[0462] 1-(3-Chloro-phenylmethanesulfonyl)-3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 9 was prepared using the same protocol as described in Example 9, substituting 8-quinoline-sulfonyl chloride with (3-chlorophenyl)-methanesulfonyl chloride.
MS(ESI) [M+H+]+=443.3.

Example 12: Synthesis of 1-(Biphenyl-4-sulfonyl)-3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 10 /
O

O

N N
S=0 I~ O

i [0463] 1-(Biphenyl-4-sulfonyl)-3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 10 was prepared using the same protocol as described in Example 9, substituting 8-quinoline sulfonyl chloride with biphenyl-4-sulfonyl chloride. The crude material was purified by preparative TLC (50% ethyl acetate: hexanes). The compound was eluted from silica with ethyl acetate and concentrated to provide an oil. The oil was washed with a minimum of methanol and a white precipitate was collected by filtration to provide 10.
MS(ESI) [M+H+]+
471Ø

Example 13: Synthesis of 3-(3,4-Dimethoxy-phenyl)-1-(naphthalene-2-sulfonyl)-pyrrolo[2,3-b]pyridine 11 /
O

O

I
N N
S=0 I O

[0464] 3-(3,4-Dimethoxy-phenyl)-1-(naphthalene-2-sulfonyl)-1H-pyrrolo[2,3-b]pyridine 11 was prepared using the same protocol as described in Example 9, substituting 8-quinoline sulfonyl chloride with 2-naphthalene-sulfonyl chloride. The crude material was purified by preparative TLC (50% ethyl acetate: hexanes). The compound was eluted from silica with ethyl acetate and concentrated to provide an oil. The oil was washed with a minimuin of methanol and a white precipitate was collected by filtration to provide 11.
MS(ESI) [M+H+]+
445Ø

Example 14: Synthesis of 3-(3,4-Dimethoxy-phenyl)-1-(4-methyl-naphthalene-l-sulfonyl)-1H-pyrrolo[2,3-b]pyridine 12 e O
o I
N N
S=0 I~ O

[0465] 3-(3,4-Dimethoxy-phenyl)-1-(4-methyl-naphthalene-l-sulfonyl)-1H-pyrrolo[2,3-b]pyridine 12 was prepared using the same protocol as described in Example 9, substituting 8-quinoline sulfonyl chloride with 4-methyl-naphthalene-l-sulfonyl chloride. The crude oil was washed with ethyl acetate, which provided a solid that was washed with acetonitrile: water and methanol: methylene chloride to provide 12. MS(ESI) [M+H+]+ = 458.9.

Example 15: Synthesis of 8-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-sulfonyl]-3-methyl-quinoline 13 e O
o N N
S=0 I O
N

[0466] 8-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-sulfonyl]-3-methyl-quinoline 13 was prepared using the same protocol as described in Example 9, substituting 8-quinoline sulfonyl cllloride with 3-methyl-quinoline-8-sulfonyl chloride. The crude oil was washed with ethyl acetate, which provided a solid that was washed with acetonitrile, methanol and methylene chloride to provide 13. MS(ESI) [M+H+]+ = 460Ø

Example 16: Synthesis of 5-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-sulfonyl]-isoquinoline 14 /
O

O

N N
S=0 O

N

[0467] 5-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-sulfonyl]-isoquinoline 14 was prepared using the same protocol as described in Example 9, substituting 8-quinoline sulfonyl chloride with isoquinoline-5-sulfonyl chloride. The crude mixture was purified by preparative TLC in two subsequent runs of 75% ethyl acetate: hexanes and 50% ethyl acetate: hexanes to provide 14. MS(ESI) [M+H+]+ = 446Ø

Example 17: Synthesis of 3-(3,4-Dimethoxy-phenyl)-1-(4-phenoxy-benzenesulfonyl)-1H-pyrrolo[2,3-b]pyridine 15 i O
o ~=

N N
' S:O
O I~

i~
~
[0468] 3-(3,4-Dimethoxy-phenyl)-1-(4-phenoxy-benzenesulfonyl)-1H-pyrrolo[2,3-b]pyridine 15 was prepared using the same protocol as described in Example 9, substituting 8-quinoline sulfonyl chloride with 4-phenoxy-benzenesulfonyl chloride. The crude mixture was crystallized from acetonitrile to provide 15. MS(ESI) [M+H+]+ = 487.1.

Example 18: Synthesis of 3-(3,4-Dimethoxy-phenyl)-1-(naphthalene-1-sulfonyl)-pyrrolo[2,3-b]pyridine 16 /
O

0 = /

N N
S=0 I O

[0469] 3-(3,4-Dimethoxy-phenyl)-1-(naphthalene-1-sulfonyl)-1H-pyrrolo[2,3-b]pyridine 16 was prepared using the same protocol as described in Example 9, substituting 8-quinoline sulfonyl chloride with 1-naphthalenesulfonyl chloride. MS(ESI) [M+H+]+ =
445.5.
Example 19: Synthesis of 1-(4-chloro-phenylmethanesulfonyl)-3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 17 Scheme - 10 /
O

O 0 =/

0 = / -~ ~ / \
N N
N 'N S'O
H O
=~
CI

Preparation of 1-(4-chloro phenylmethanesulfonyl)-3-(3,4-dimetlaoxy phenyl)-1H-pyrrolo[2,3-b]pyridine 17 [0470] 3-(3,4-Dimethoxy-phenyl)-1H-pyrrolo-[2,3-b]pyridine 1(40 mg, 0.16 mmol) was dissolved in tetrahydrofuran (3.5 mL). Sodium hydride (60% dispersion in mineral oil, 10 mg, 0.24 mmol) was added. The reaction was stirred for 15 minutes. Into the reaction mixture was added (4-chloro-phenyl)-methanesulfonyl chloride (40 mg, 0.18 mmol). The mixture was stirred overnight, concentrated, and partitioned between brine and ethyl acetate. The organic portions was dried with anhydrous sodium sulfate, filtered, and concentrated.
The product was purified by reverse phase HPLC with 40-100% acetonitrile: 0.1 % aqueous formic acid. The appropriate fractions were lyophilized to provide 17 as the formate salt (7.5 mg, 11 %).
MS(ESI) [M+H+]+ = 443Ø

Example 20: Synthesis of 3-(3,4-dimethoxy-phenyl)-1-(3-nitro-phenylmethanesulfonyl)-1H-pyrrolo[2,3-b]pyridine 18 /
O

O
=

N N
S-'~ O
*O

[0471] 3-(3,4-Dimethoxy-phenyl)-1-(3-nitro-phenylmethanesulfonyl)-1H-pyrrolo[2,3-b]pyridine 18 was prepared using the same protocol as described in Example 19, substituting (4-chloro-phenyl)-methanesulfonyl chloride with (3-nitropheny)-methanesulfonyl chloride.
The crude material was purified by preperative TLC (50% ethyl acetate:
hexanes). MS(ESI) [M+H+]+ = 454Ø

Example 21: Synthesis of 1-benzenesulfonyl-3-(3,4-dimethoxy-phenyl)-1H-indole Scheme - 11 /
O

Br O
=
/ ~, -~ / = ~
N
O"S N
O' ~ ~ / = O
Q

[0472] 1-Benzenesulfonyl-3-bromoindole 20 (350 mg, 1.0 mmol) was dissolved in tetrahydrofuran (6 mL). Into the solution was added 3,4-dimethoxyphenyl boronic acid (379 mg, 2.1 mmol), tetrakis(triphenylphosphine)palladium(0) (60 mg, 0.05 mmol) and potassium carbonate (3 mL). The reaction mixture was heated in a CEM Discover microwave instrument at 120 C for 10 minutes. The reaction mixture was concentrated under reduced pressure and partioned between ethyl acetate and saturated sodium bicabonate.
The organic portion was dried over anhydrous magnesium sulfate, and filtered. The filtrate was adsorbed onto silica gel and purified by flash chromatography (5%-50% ethyl acetate:
hexanes) to provide the desired product, 19, (359 mg, 88%).

Example 22: Synthesis of 8-[3-(3,4-dimethoxy-phenyl)-indole-l-sulfonyl]-quinoline 21 Scheme -12 /

O

O = /
~ = / Step 2 Step 1 N N

Step 1-Preparation of 3-(3,4-dimetho.xy phenyl)-indole 22 [0473] 1-Benzenesulfonyl-3-(3,4-dimethoxy-phenyl)-indole (350 mg, 0.89 mmol) 19 was dissolved in ethanol (3 mL) and potassium hydroxide pellets (385 mg, 6.9 mmol) were added.
The reaction mixture was heated in a CEM Discover microwave instrument at 120 C for 10 minutes. The reaction mixture was concentrated to dryness and the residue was partitioned between ethyl acetate and saturated sodium bicarbonate. The organic portion was dried with anhydrous magnesium sulfate, filtered and the filtrate concentrated to provide the desired product 22,which was used without fiuther purification (208 mg, 92 %).

Step 2- Preparation of 8-[3-(3,4-dimethoxy phenyl)-indole-l-sulfonylJ-quinoline 21 [0474] 3-(3,4-Dimethoxy-phenyl)-indole 22 (70 mg, 0.28 mmol) was dissolved in methylene chloride (5.0 mL). Tetra-N-butylammonium bromide (3 mg, 0.01 mmol) and aqueous potassium hydroxide (50% wt/vol, 500 L) were added. The reaction mixture was stirred for 5 minutes. 8-quinoline-sulfonyl chloride (89.5 mg, 0.39 mmol) was added and the reaction mixture was stirred for 2 hours. The product was extracted with 2 N lithium hydroxide and methylene chloride. The organic portion was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated and purified by flash chromatography (ethyl acetate:
hexanes 0%-50%). The appropriate fractions were combined and concentrated to provide 21, (76.1 mg, 90 %). MS(ESI) [M+H+]+= 445Ø

Example 23: Synthesis of 8-[3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine -1-sulfonyl]-3-methyl-quinoline 24 Scheme - 13 /

CI. ~ ~

N N

le, S'O

Preparation of 8-[3-(3, 4-dimetlaoxy phenyl) pyrrolo[2, 3-bJpyf-idine-l-sulfonylJ-3-methyl-quinoline 24 [0475] 3-(3,4-Dimethoxy-phenyl)-1H-pyrrolo-[2,3-b]pyridine,1, (40 mg, 0.1 mmol) was dissolved in methylene chloride (4 mL). Aqueous potassium hydroxide (50%
wt/vol, 300 L) and tetrabutylammonium hydrogen sulfate (20 mg, 0.007 mmol) were added. The reaction mixture was stirred for 10 minutes at room temperature. Into the reaction was added 6-methyl-quinoline-8-sulfonyl chloride (68 mg, 0.28 mmol), prepared as described (Lubenets, V. I.;
Stadnitskaya, N. E.; Novikov, V. P.; Russ. J. Org. Chena.; 36; 2000; 851 -853) and the reaction mixture was stirred for 2 hours at room temperature. The reaction mixture was concentrated and the residue was washed with brine and ethyl acetate. The organic portion was dried with anhydrous sodium sulfate and filtered. The filtrate was concentrated and the resulting solid was washed with acetonitrile to provide 24 (43 mg, 60 %).
MS(ESI) [M+H+]+=460.1.

Example 24: Synthesis of 8-[3-(3,4-dimethoxy-phenyl)-indole-l-sulfonyl]-3-methyl-quinoline 25 /
O

0 = /

N
~ = S:O
~ Q
N
=~

[0476] 8-[3-(3,4-Dimethoxy-phenyl)-indole-l-sulfonyl]-3-methyl-quinoline 25 was prepared using the same protocol as described in Example 22, substituting 8-quinoline-sulfonyl chloride with 6-methyl-quinoline-8-sulfonyl chloride. After the crude material was concentrated, acetonitrile was added to the oil and let stand. The resulting precipitate was filtered and further washed with acetonitrile and dried in vacuo to provide the desired product 25.
MS(ESI) [M+H+]+ = 459.1.

Example 25: Synthesis of 3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (4-chloro-phenyl)-amide 26 Scheme -14 /

O 0 = /
0 = / -~ ~ ~ ~
N N
H N H N

CI

Preparation of 3-(3,4-dimethoxy phenyl) pyrYolo[2,3-bJpyridine-l-carboxylic acid (4-chloro-phenyl)-amide 26 [0477] 3-(3,4-Dimethoxy-phenyl)-1H-pyrrolo-[2,3-b]pyridine 1(40 mg, 0.16 mmol) was dissolved in benzene and 4-chlorophenyl isocyanate (36 mg, 0.24 mol) dissolved in tetrahydrofuran (4 inL) was added. 4-Dimethylaminopyridine (20 mg, polymer bound) was added. The reaction was heated in a CEM Discover microwave at 200 C for 10 minutes. The mixture was filtered and concentrated. The resulting solid was washed with a miniinum of methanol, filtered, and then washed with dichloromethane to provide 26, (26 mg, 40 %).
MS(ESI) [M+H+]+ = 408.4.

Example 26: Synthesis of 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3-chloro-4-methoxy-phenyl)-amide 27 /
O

0 = /

N N
HN-~-O
ici "O

[0478] 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3-chloro-4-methoxy-phenyl)-amide 27 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate with 3-chloro-4-methoxyphenyl isocyanate. MS(ESI) [M+H+]+ = 43 8.4.

Example 27: Synthesis of 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid phenylamide 28 /
O

0 = /
I
N N
H N'-.kl O

[0479] 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid phenylamide 28 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate with phenylisocyanate. After an initial heating at 120 C for 15 minutes, additional phenyl isocyanate was added (500 gL) and the reaction was heated again in the microwave for 30 ininutes at 180 C. The concentrated solid was washed with a minimum of acetonitrile and purified by reverse phase HPLC (0.1% formic acid: acetonitrile) to provide the desired product 28. MS(ESI) [M+H+]+ = 374.3.

Example 28: Synthesis of 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3,5-dimethoxy-phenyl)-amide 29 I
O

O
=

N N
HN-11~O
O O' [0480] 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3,5-dimethoxy-phenyl)-amide 29 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate with 3,5-dimethoxyphenyl isocyanate and microwave heating for 20 minutes at 200 C. MS(ESI) [M+H+]+ = 434.2.

Example 29: Synthesis of 3-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carbonyl]-ainino-benzoic acid ethyl ester 30 =
O

O
=

N N
H N'1~O

O
611~' O~
[0481] 3-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carbonyl]-amino-benzoic acid ethyl ester 30 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate witli 3-isocyanato-benzoic acid ethyl ester and microwave heating for 20 minutes at 200 'C. After washing with a minimum of methanol and methylene chloride, the reaction mixture was diluted with methanol: methylene chloride (95:5) and let stand. The resulting precipitate was isolated by filtration and dried in vacuo to provide 30. MS(ESI) [M+H+]+ = 446.5.

Example 30: Synthesis of 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (4-fluoro-3-nitro-phenyl)-amide 31 /
O
O

N N
H N',:~O
~I

F

[0482] 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (4-fluoro-3-nitro-phenyl)-amide 31 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate with 1-fluoro-4-isocyanato-2-nitro-benzene and reacting at room temperature for two hours. The resulting precipitate was washed with methanol and dichloromethane and dried in vacuo to provide 31. MS(ESI) [M+H+]+
= 437.5.
Example 31: Synthesis of 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3-methoxy-phenyl)-amide 32 /
O

O

N N
HN'k-p [0483] 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3-methoxy-phenyl)-amide 32 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate with 3-methoxy-phenyl isocyanate. After filtration and concentration, the reaction mixture was redissolved in acetonitrile and a minimum of water and let stand. The resulting precipitate was collected by filtration and dried in vacuo to provide 32. MS(ESI) [M+H+]+ = 404.4.

Example 32: Synthesis of 4-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carbonyl]-amino-benzoic acid ethyl ester 33 /
O

O = /

I
N N
HN-11:~O
O O

[0484] 4-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carbonyl]-amino-benzoic acid ethyl ester 33 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate with 4-isocyanato-benzoic acid ethyl ester.

Example 33: Synthesis of 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3-acetyl-phenyl)-amide 34 /
O

O = /

N N
HN-11~O
tk--f [0485] 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3-acetyl-phenyl)-amide 34 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate with 3-acetyl-phenyl isocyanate.
MS(ESI) [M+H+]+ _ 416.3.

Example 34: Synthesis of 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid(3-fluoro-phenyl)-amide 35 /
O

O

N N
HN-11~O
~I
~ F
[0486] 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid(3-fluoro-phenyl)-amide 35 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate with 3-fluoro-isocyanato-benzene.
MS(EST) [M+H+]+ _ 392Ø

Example 35: Synthesis of 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acidbenzo[1,3]dioxol-5-ylamide 36 /
O

O

o s~ N N
HN'k-O
O

[0487] 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acidbenzo[1,3]dioxol-5-ylamide 36 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate with 5-isocyanato-benzo[1,3]dioxole.
After the initial microwave heating, the reaction was heated again in the microwave for 10 minutes at 210 C. The crude material was recrystallized from methanol:
methylene chloride to provide 36. MS(ESI) [M+H+]+ = 418.5.

Example 36: Synthesis of 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3,4 -dichloro-phenyl)-amide 37 /
O

O
=

N N
H N'k, O

CI
CI

[0488] 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3,4 -dichloro-phenyl)-amide 37 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate with 1,2-dichloro-4-isocyanato-benzene.
MS(ESI) [M+H+]+ = 443.5.

Example 37: Synthesis of 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3-nitro-phenyl)-amide 38 /
O

O
=~
N N
HN---K- O

[0489] 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3-nitro-phenyl)-amide 38 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate with 1-isocyanato-3-nitro-benzene. After the initial microwave heating, the reaction was heated again in a CEM Discover microwave for 6 minutes at 200 C
with an additional equivalent of 1-isocyanato-3-nitro-benzene. MS(ESI) [M+H+]+
= 419.2.
Example 38: Synthesis of 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (4-methoxy-phenyl)-amide 39 /
O

O
=~
N N
HN-k-O
~I

~O

[0490] 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (4-methoxy-phenyl)-amide 39 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate with 4-methoxyphenyl isocyanate. After the initial microwave heating, the reaction was heated again in the microwave for 5 minutes at 190 C.
MS(ESI) [M+H+]+ = 404.2.

Example 39: Synthesis of 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (4-acetyl-phenyl)-amide 40 i O
0 =/
=~
N N

O
[0491] 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (4-acetyl-phenyl)-amide 40 was prepared using the same protocol as described in Example 25, substituting 4-chlorophenyl isocyanate with 4-acetylphenyl isocyanate. After the initial microwave heating, the reaction was heated again in the microwave for 20 minutes at 220 C.
MS(ESI) [M+H+]+ = 416.2.

Example 40: Synthesis of 3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carbothioic acid phenylamide 41 Scheme - 15 =
O

O O
0 ~ / -~_ ~ = 1 / = ~ N N
H N HN

1 i~
~

Preparation of 3-(3,4-dinaetlzoxy plzenyl) pyrrolo[2,3-b]pyridine-l-carbothioic acid phenylamide 41 [0492] 3-(3,4-Dimethoxy-phenyl)-1H-pyrrolo-[2,3-b]pyridine 1(50 mg, 0.20 mmol) was dissolved in N,N-dimethylformamide (4 mL). Sodium hydride (10 mg, 0.24 mmol, 60%

dispersion in mineral oil) was added. The reaction mixture was stirred for 15 minutes at room temperature. 1-Isothiocyanatobenzene (35 L, 0.29 mmol) was added a.nd the reaction mixture was stirred for 40 minutes at room temperature. The product was concentrated and the residue was partitioned between brine and ethyl acetate. The organic portion was dried over anhydrous sodium sulfate, filtered, and concentrated to provide a solid. The solid was washed with a minimum of methanol and dried in vacuo to provide 41, (38 mg, 49 %).
MS(ESI) [M+H+]+ = 390.2.

Example 41: 3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carbothioic acid (3-methoxy-phenyl)-ainide 42 /
Q

O

N N
HN'k**S

[0493] 3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carbothioic acid (3-methoxy-phenyl)-amide 42 was prepared using the same protocol as described in Example 40, substituting 1-isothiocyanatobenzene with 1-chloro-4-isothiocyanatobenzene.

Example 42: Synthesis of 8-[3-(2-methoxy-pyrimidin-5-yl)-pyrrolo[2,3-b]pyridine-l-sulfonyl]-quinoline 43 Scheme - 16 O
Br Br N y Stepl N Step2 QN- ~ N
00- N N N NO N t H 0,S O:S:O
N WN--Step-I Preparation of 8-(3-bronao pyrrolo[2,3-bJpyf idine-l-sulfonyl)-quinoline 44 [0494] Into a round bottom flask was added 3-bromo-7-azaindole (3, 1.18 g, 5.99 mmol) and tetra-N-butylammonium bromide (193 mg, 0.600 mmol), and 5.0 M sodium hydroxide (15.4 mL). 8-Quinoline-sulfonyl chloride (1.64 g, 7.19 mmol) in dichloromethane (5.9 mL) was added dropwise at room temperature. After a few hours, all starting materials were gone.
After 30 mL of dichlormethane was added, two layers were separated. The aqueous layer was washed with dichloromethane. The combined organic layers were washed with 1M
sodium bicarbonate, water, and brine and dried over anhydrous sodium sulfate. The crude material was concentrated under reduced pressure and was purified by colunm chromatography (55 - 80%
ethyl acetate in hexane) to yield the desired product as a light yellow colored solid (44, 1.72 g, 4.43 mmol). MS(ESI) [M+Ff' ]+ = 389.4.

Step-2 Preparation of 8-[3-(2-fnethoxy pyrifnidin-5 yl) pyrrolo[2,3-bJpyridine-l-sulfonylJ-quinoline 43 [0495] In a microwave reaction tube, 8-(3-bromo-pyrrolo[2,3-b]pyridine-l-sulfonyl)-quinoline (44, 68 mg, 0.18 mmol), 2-methoxy-pyrimidine-4-boronic acid (67.4 mg, 0.438 mmol), and tetrakis(triphenylphosphine)palladium(0) (10 mg, 0.0088 mmol) were mixed in 1.0 M of potassium carbonate (0.52 mL) and tetrahydrofuran (0.84 mL). The resulting mixture was heated at 120 C in a CEM Discover microwave unit for 10 minutes. Ethyl acetate and water were added and two layers were separated. The aqueous layer was extracted with ethyl acetate and the combined organic layers were washed with brine and dried over sodium sulfate.
The product was concentrated under reduced pressure and the resultant crude material was purified by column chromatography (80 - 90% ethyl acetate in hexane) to yield the desired product as a white solid (43, 0.005 g, 0.01 mmol). MS(ESI) [M+H+]+ = 417.8.

Example 43: Synthesis of 3-(2-Methoxy-pyrimidin-5-yl)-1-phenylmethanesulfonyl-lH-pyrrolo[2,3-b]pyridine 45 NYO
QN-\ ~N N

O:S=O
=~

[0496] 3-(2-Methoxy-pyrimidin-5-yl)-1-phenylmethanesulfonyl-1H-pyrrolo[2,3-b]pyridine 45 was prepared using the same protocol as described in Example 42, substituting 8-quinoline-sulfonyl chloride with phenyl methanesulfonyl chloride. MS(ESI) [M+H+]+ =
381.2.

Example 44: Synthesis of 8-[3-(2-Methylsulfanyl-pyrimidin-4-yl)-pyrrolo[2,3-b]pyridine-l-sulfonyl]-quinoline 46 Scheme - 17 4~
O, p NNS
Br B, Step 1 Step 2 N S p N -S ~ N N. O
~~ 0*/ = 0 / =
N N N
, - - , Step-1 PrepaNation of 8-[3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2 yl) pyYrolo[2,3-bJpyridine-l-sulfonylJ-quinoline 47 [0497] Into a round bottom flask, 8-(3-bromo-pyrrolo[2,3-b]pyridine-l-sulfonyl)-quinoline (44, 335 mg, 0.863 mmol), diboron pinacol ester (263 mg, 1.04 mtnol), and bis(triphenylphosphine)palladium(II) chloride (18 mg, 0.026 mmol) were added.
Under an atmosphere of nitrogen, N,N-dimethylformamide (4.0 mL) was added. The mixture was thoroughly degassed by alternately connecting the flask to vacuum and nitrogen source. The resulting mixture was heated to 100 C overnight. After the reaction mixture was cooled to room temperature, it was poured into water and extracted with ethyl acetate twice. The combined organic layers were washed with water and brine and dried over anhydrous sodium sulfate. The product was concentraqted under reduced pressure and the crude material was purified by column chromatography (55% ethyl acetate in hexane) to yield the desired product as a white solid (47, 70 mg, 0.16 mmol). MS(ESI) [M+H+]+ = 435.8.

Step-2 Preparation of 8-[3-(2Methylsulfanyl pyt=imidin-4 yl) pyrf=olo[2,3-b]pyidine-1-sulfonylJ-quinoline 46 [0498] In a microwave reaction tube, 8-[3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyrrolo [2,3-b]pyridine- 1 -sulfonyl] -quinoline (47, 70 mg, 0.16 mmol), 4-iodo-2-metllylsulfanyl-pyrimidine (101 mg, 0.402 mmol), and tetrakis(triphenylphosphine)palladium(0) (9.3 mg, 0.008 mmol) were mixed in 1.0 M
potassium carbonate in water (0.48 mL) and tetrahydrofuran (0.77 mL). The resulting mixture was heated at 120 C in a CEM Discover microwave unit for 10 minutes. Ethyl acetate and water were added, and the two layers were separated. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine and dried over sodium sulfate. Concentration under reduced pressure afforded the crude material, which was purified by column chromatography (50% ethyl acetate in hexane) to yield the desired product in light yellow solid (46, 10 mg, 0.023 mmol). MS(ESI) [M+H+]+ = 434Ø

Example 45: Synthesis of [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-ethoxy-napllthalen-1-yl)-methanone 52 Scheme - 18 O O O O
N 4~~ N
H N N _ ~~

52 ~

[0499] 3-(3,4-Dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 1 (638 mg, 2.51 inmole) was dissolved in DMF (30 mL) and sodium hydride (60 % dispersion in mineral oil, 100 mg, 2.50 mmole) was added in small portions to the reaction mixture. After stirring for 30 minutes, 2-ethoxy-1-naphthoyl chloride (646 mg, 2.76 mmole) was added and the reaction was stirred for 4 hours. The reaction mixture was poured into 200 mL of ice water and was extracted with ethyl acetate. The organic layer was washed with saturated bicarbonate solution followed by saturated potassium hydrogen sulfate and then brine. The resulting solution was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure.
The residue was purified by silica gel chromatography (30 % ethyl acetate: hexanes) to yield 800 mg (70%) of the titled compound as a white solid MS(ESI) [M+H+]+ = 453.06.

Example 46: Synthesis of 3-(3,4-Dimethoxy-phenyl)-1-(3-nitro-benzyl)-1H-pyrrolo[2,3-b]pyridine 53 Scheme - 19 O
= /=
~I ' -~~ ~~ =

N H ~
~~

[0500] 3-(3,4-Dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 1 (35 mg, 0.14 mmole) was dissolved in 15 mL of DMF and sodium hydride (60 % dispersion in mineral oil, 10 mg, 0.25 mmol) was added in small portion to the reaction mixture. After stirring for 30 minutes, m-nitro-benzyl chloride (30 mg, 0.14 mmole) was added to the reaction mixture and was stirred for 2 hours. The reaction mixture was poured into 50 mL of ice water and was extracted with ethyl acetate. The organic layer was washed with saturated sodium bicarbonate followed by saturated potassium bisulfate and then brine. The resulting solution was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (30 % ethyl acetate in hexanes) to yield 42 mg (71%) of the titled coinpound as a white solid. MS(ESI) [M+H+]+ = 390.1.

Example 47: Synthesis of [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-(2-ethoxy-4-nitro-phenyl)-methanone 54 Scheme - 20 =

/= /=

; I ~ = ~ ~ O
N H N N

[0501] 3-(3,4-Dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 1 (250 mg, 0.98 mmole) was dissolved in 50 mL of CH2C12. N,N-diethylisopropylamine (205 uL, 1.2 mmol), 2-ethoxy-4-nitro-benzoic acid (228 mg, 1.1 mmol) and bromotris(pyrolodino)phoshonium hexafluorophosphate (550 mg, 1.1 mmol) were added to the stirring reaction mixture. The mixture was stirred at ambient temperature for 4 h and washed consecutively with saturated sodium bicarbonate and saturated potassium bisulfate solution. The resulting solution was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure.
The residue was purified by silica gel chromatography (30 % ethyl acetate in hexanes) to yield 340 mg (77%) of the titled compound as a yellow solid. MS(ESI) [M+H+]+ =
448.2.
Example 48: Synthesis of [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]phenylmethanone-amide 55 ~ = O
IA N =
N N
~
o ~s [0502] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]phenylmethanone-amide 55 was prepared using the saine protocol as described for Example 45, substituting 2-ethoxy-1-naphthalene carbonyl chloride with benzoyl chloride. MS(ESI) [M+H+]+ = 359.2.

Example 49: Synthesis of [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(3-pyridylyl)-methanone - amide 56 O.-/ = O' ~ =
N'o N
O
N

[0503] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-(3-pyridylyl)-methanone-amide 56 was prepared using the same protocol as described for Example 45, substituting 2-ethoxy-1-naphthalene carbonyl chloride with pyridine-3-carbonyl chloride.
MS(ESI) [1VI+H+]+ = 360Ø

Example 50: Synthesis of [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-(4-trifluoromethypyridn-3-yl)-methanone - amide 57 O.-= O' F NF N F O

N

[0504] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(4-trifluoromethypyridn-3-yl)-methanone - ainide 57 was prepared using the same protocol as described for Example 45 substituting 2-ethoxy-1-naphthalene carbonyl chloride with 4-trifluoromethylpyridine-3-carbonyl chloride. MS(ESI) [M+H+]+ = 428Ø

Example 51: Synthesis of [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-(l-naphthyl)-methanone - amide 58 O.-= O' N; N O

[0505] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-(1-naphthyl)-methanone amide 58 was prepared using the same protocol as described for Example 45, substituting 2-ethoxy-1-naphthalene carbonyl chloride with naphthylene-1-carbonyl chloride.
MS(ESI) [M+H+]+ = 409.1.

Example 52: Synthesis of [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-(2-chloropyridn-3-yl)-methanone - amide 59 O.-= O
N N _ ~
O N
ci [0506] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-chloropyridn-3-yl)-methanone - amide 59 was prepared using the same protocol as described for Example 45, substituting 2-ethoxy- 1 -naphthalene carbonyl chloride with 2-chloropyridine-3-carbonyl chloride. MS(ESI) [M+H+]+ = 393.9.

Example 53: Synthesis of 1-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-2-(4-methoxy-phenyl)-ethanone - amide 60 O.-= O' I
N N
O
O
=
[0507] 1-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-2-(4-methoxy-phenyl)-ethanone - amide 60 was prepared using the same protocol as described for Example 45, substituting 2-ethoxy-l-naphthalene carbonyl chloride with (4-methoxy-phenyl)-acetyl chloride. MS(ESI) [M+H+]+ = 403Ø

Example 54: Synthesis of [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-(2-ethoxy-phenyl)-methanone - amide 61 O.-/
~ = O
~ ~ =
N N _ ~s /-O

[0508] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-ethoxy-phenyl)-methanone - amide 61 was prepared using the same protocol as described for Example 45 substituting 2-ethoxy-l-naphthalene carbonyl chloride with 2-ethoxybenzoyl chloride. MS(ESI) [M+H+]+
403.5.

Example 55: Synthesis of Acetic acid 6-[3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carbonyl]-naphthalen-1-yl ester - amide 62 O.-/
~ = O
~ ~ =
N N
~ ~
OO

[0509] Acetic acid 6-[3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carbonyl]-naphthalen-1-yl ester - amide 62 was prepared using the same protocol as described for Example 47, substituting 2-ethoxy-4-nitrobenzoic acid with 5-acetoxy-naphthalene-2-carboxylic acid. MS(ESI) [M+H+]+= 467.5.

Example 56: Synthesis of Benzo[b]thiophen-3-yl-[3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl] -methanone 63 O~
/
a-N N p O S

[0510] Benzo[b]thiophen-3-yl-[3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-methanone 63 was prepared using the same protocol as described for Example 45, substituting 2-ethoxy-l-naphthalene carbonyl chloride with benzo[b]thiophene-3-carbonyl chloride.
Example 57: Synthesis of [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(4-fluoro-naphthalen-l-yl)-methanone 64 O.-/ = O
~ ~ =
N N

O F

[0511] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(4-fluoro-naphthalen-l-yl)-methanone 64 was prepared using the same protocol as described for Example 47, substituting 2-etlioxy-4-nitrobenzoic acid with 4-fluro-naphthalene-l-carboxylic acid.
MS(ESI) [M+H+]+
427.5.

Example 58: Synthesis of [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-(5-methoxynaphthalen-1-yl)-methanone 65 0.-= O
~ ~ =
N N _ ~ ~
O

~ 0 [0512] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(5-methoxynaphthalen-l-yl)-methanone 65 was prepared using the same protocol as described for Example 47, substituting 2-ethoxy-4-nitrobenzoic acid with 5-methoxy-naphthalene-2-carboxylic acid.

Example 59: Synthesis of [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-isoquinolin-8-yl-methanone 66 0.-~ = O
I ; =
N N

O TN/

[0513] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-isoquinolin-8-yl-methanone 66 was prepared using the same protocol as described for Example 47, substituting 2-ethoxy-4-nitrobenzoic acid with quinolin-8-carboxylic acid. MS(ESI) [M+H+]+ = 410.5.

Example 60: Synthesis of 1-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-ethanone 67 / = 0 ~ ~ =
N ~
O

[0514] 1-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-ethanone 67 was prepared using the same protocol as described for Example 45, substituting 2-ethoxy-l-naphthalene carbonyl chloride with acetyl chloride. MS(ESI) [M+H+]+ = 279.2.

Example 61: Synthesis of [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-(2-methoxyphenyl)-methanone 68 =
O
= O
N N
_ O
~a O
X

[0515] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-methoxyphenyl)-methanone 68 was prepared using the same protocol as described for Example 45, substituting 2-ethoxy-1-naphthalene carbonyl chloride with 2-methoxybenzoyl chloride.
MS(ESI) [M+H+]+
= 389.15.

Example 62: Synthesis of 3-(3,4-Dimethoxy-phenyl)-1-[4-(3-methyl-pyridin-4-yl)-benzenesulfonyl]-1H-pyrrolo[2,3-b]pyridine 69 0.-/ = O' ~ ~ =
N N

N

[0516] 3-(3,4-Dimethoxy-phenyl)-l-[4-(3-methyl-pyridin-4-yl)-benzenesulfonyl]-pyrrolo[2,3-b]pyridine 69 was prepared using the same protocol as described in Example 9, substituting 8-quinoline-sulfonyl chloride with 4-(3-methyl-pyridin-4-yl)-benzenesulfonyl chloride. MS(ESI) [M+H+]+= 487Ø

Example 63: Synthesis of 3-(3,4-Dimethoxy-phenyl)-1-(4-oxazol-5-yl-benzenesulfonyl)-1H-pyrrolo[2,3-b]pyridine 70 0 .-/ = 0' N N O
p== G01 ~ ~N
[0517] 3-(3,4-Dimethoxy-phenyl)-1-(4-oxazol-5-yl-benzenesulfonyl)-1H-pyrrolo[2,3-b]pyridine 70 was prepared using the same protocol as described in Example 9, substituting 8-quinoline-sulfonyl chloride with 4-oxazol-5-yl-benzenesulfonyl chloride.
MS(ESI) [M+H+]+=
462Ø

Example 64: Synthesis of 3-(3,4-Dimethoxy-phenyl)-1-(4-pyrazol-1-yl-benzenesulfonyl)-1H-pyrrolo[2,3-b]pyridine 71 0.-~ = 0 ~ ~ =
N N
O ~

N~

[0518] 3-(3,4-Dimethoxy-phenyl)-1-(4-pyrazol-1-yl-benzenesulfonyl)-1H-pyrrolo[2,3-b]pyridine 71 was prepared using the same protocol as described in Example 9, substituting 8-quinoline-sulfonyl chloride with 4-pyrazol-1 -yl-benzenesulfonyl chloride.
MS(ESI) [M+H+]+_ 461Ø

Example 65: Synthesis of 3-(3,4-Dimethoxy-phenyl)-1-(3-oxazol-5-yl-benzenesulfonyl)-1H-pyrrolo[2,3-b]pyridine 72 ~ = O' N

[0519] 3-(3,4-Dimethoxy-phenyl)-1-(3-oxazol-5-yl-benzenesulfonyl)-1H-pyrrolo[2,3-b]pyridine 72 was prepared using the same protocol as described in Example 9, substituting 8-quinoline-sulfonyl chloride with 3-oxazol-5-yl-benzenesulfonyl chloride.
MS(ESI) [M+H+]+_ 462Ø

Example 66: Synthesis of 3-(3,4-Dimethoxy-phenyl)-1-[4-(5-methyl-[1,3,4]oxadiazol-2-yl)-benzenesulfonyl]-1H-pyrrolo[2,3-b]pyridine 73 =

~ ~ =
N N O

O:S ' X O N

[0520] 3-(3,4-Dimethoxy-phenyl)-1-[4-(5-methyl-[1,3,4]oxadiazol-2-yl)-benzenesulfonyl]-1H-pyrrolo[2,3-b]pyridine 73 was prepared using the same protocol as described in Example 9, substituting 8-quinoline-sulfonyl chloride with 3-oxazol-5-yl-benzenesulfonyl chloride.
MS(ESI) [M+H+]+ = 477Ø

Example 67: Synthesis of 1-(2-Benzenesulfonylmethyl-benzyl)-3-(3,4-diinethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 74 O' i~ O%
-.
N N
O.S .O

[0521] 1-(2-Benzenesulfonylmethyl-benzyl)-3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 74 was prepared using the same protocol as described in Example 46, substituting in-nitro-benzyl chloride with 1-benzenesulfonylmethyl-2-chloromethyl-benzene.
MS(ESI) [M+H+]+= 499.1.

Example 68: Synthesis of 1-Benzyl-3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 75 ~
O
O, N N

[0522] 1-Benzyl-3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 75 was prepared using the same protocol as described in Example 46, substituting m-nitro-benzyl chloride with benzyl chloride. MS(ESI) [M+H+]+= 345.1.

Example 69: Synthesis of 3-(3,4-Dimethoxy-phenyl)-1-(2-methyl-naphthalen-1-ylmethyl)-1H-pyrrolo[2,3-b]pyridine 76 O.-1 = O' N N
~
~

[0523] 3-(3,4-Dimethoxy-phenyl)-1-(2-methyl-naphthalen-1-ylmethyl)-1H-pyrrolo[2,3-b]pyridine 76 was prepared using the same protocol as described in Example 46, substituting m-nitro-benzyl chloride with 1-chloromethyl-2-methyl-naphthalene.

Example 70: Synthesis of 4-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-ylmethyl]-benzonitrile 77 O
N N

[0524] 4-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-ylmethyl]-benzonitrile 77 was prepared using the same protocol as described in Example 46, substituting m-nitro-benzyl chloride witli 4-chloromethylbenzonitrile. MS(ESI) [M+H+]+= 370.2.

Example 71: Synthesis of 1-Biphenyl-2-ylmethyl-3-(3,4-dimethoxy-phenyl)-IH-pyrrolo[2,3-b]pyridine 78 O' 01, N N
~
~o [0525] 1-Biphenyl-2-ylmethyl-3-(3,4-dimethoxy-phenyl)-IH-pyrrolo[2,3-b]pyridine 78 was prepared using the same protocol as described in Example 46, substituting m-nitro-benzyl chloride with 2-phenylbenzyl chloride.

Example 72: Synthesis of 3-(3,4-Dimethoxy-phenyl)-1-(3-trifluoromethyl-benzyl)-pyrrolo[2,3-b]pyridine 79 1;' F F
F

[0526] 3-(3,4-Dimethoxy-phenyl)-1-(3-trifluoromethyl-benzyl)-1H-pyrrolo[2,3-b]pyridine 79 was prepared using the same protocol as described in Example 46, substituting m-nitro-benzyl chloride with m-trifluoromethylbenzyl chloride. MS(ESI) [M+H+]+= 413Ø
Example 73: Synthesis of compounds of Formula lb Scheme - 21 CI A A
_W -W _w u jx Stepl u x Step2 ~ x t'H y t~ H y u~ Y
tN
~ $ k XI XII
Formula lb Step -1 Synthesis of Formula XII
[0527] Compound XI can be prepared from 7-azaindole following published procedure (Schneller, S. W.; Luo, Jiann-Kuan. J. Org. Claem. 1980, 45, 4045-4048), and compound of Formula XII can prepared from compound XI by reacting with boronic acids under Suzuki reaction conditions (aqueous base and Pd(0) catalyst or anhydrous conditions with KF in dioxane with Pd(0) catalyst). Alternately, compound XI can also be reacted with a tin reagent, zinc reagent or copper reagent, under Stille, Negishi or cuprate coupling reaction conditions respectively, to provide compound XII. The product can isolated by conventional work up procedure, e.g. extraction of the product with an organic solvent and purifying by column chromatography.

Step -2 Synthesis of Formula lb [0528] Compound of Formula lb can be prepared by reacting compound XII with nucleophilic reagents, e.g. acid chlorides, sulfonyl chlorides, isocyanates, isothiocyanates, alkyl halides, benzyl halides, etc., under basic conditions. The product can be isolated by following standard workup procedures, e.g. extraction of the product with organic solvent and purifying by column chromatography.

Example 74: Synthesis of [4-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-ethoxy-naphthalen-1-yl)-methanone 48 Scheme - 22 O O
O., CI
Step 1 Step 2 ~ ~ = ~ ~ = ~ ~ =

N H N H N N O

Step-I Preparation of 4-(3, 4-Dimethoxy phenyl)-1 H pyf=Yolo[2, 3-b]pyf=idine [0529] In a microwave reaction tube, 4-chloro-7-azaindole (49, 1.362 g, 8.926 mmol), prepared from 7-azaindole following published procedure (Schneller, S. W.;
Luo, Jiann-Kuan.
J. Org. Claefn. 1980, 45, 4045-4048), 3,4-dimethoxyphenyl boronic acid (4.06 g, 22.3 mmol), and tetrakis(triphenylphosphine)palladium(0) (52 mg, 0.45 mmol) were mixed in 1.0 M
potassium carbonate in water (27 mL) and tetrahydrofuran (43 mL). The resulting mixture was heated at 150 C in a CEM Discover microwave unit for 20 minutes. Ethyl acetate and water were added, and the two layers were separated. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine and dried over anhydrous sodium sulfate. Concentration under reduced pressure afforded the crude material, which was purified by column chromatography (40 - 70% ethyl acetate in hexane) to yield the desired product in light yellow solid (50, 974 mg, 3.83 mmol). MS(ESI) [M+H+]+ =
255.2.

Step-2 PYepaf-ation of [4-(3,4-Dimethoxy phenyl) pyYrolo[2,3-bJpyridin-1 ylJ-(2-ethoxy-naphtlaalen-1 yl)-methanone 48 [0530] Sodium hydride (60% dispersion in mineral oil, 39.9 mg, 0.997 mmol) was washed with hexane and put under an atmosphere of nitrogen, and 1 mL of tetrahydrofuran was added.
4-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine (50, 195 mg, 0.767 mmol) in tetrahydrofiiran (6.2 mL) was added, and the resulting mixture was stirred for 10 minutes at room temperature. 2-ethoxy naphthoyl chloride (202 mg, 0.844 mmol) in tetrahydrofuran was added. After two hours, the reaction was quenched with water, and the two layers were separated. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine and dried over anhydrous sodium sulfate. Concentration under reduced pressure afforded.the crude material, which was purified by column chromatography (35 - 60% etliyl acetate in hexane) to yield the desired product in light yellow solid (48, 262 mg, 0.579 mmol). MS(ESI) [M+H+]+ = 453.2.

Example 75: Synthesis of [4-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-ethoxy-phenyl)-methanone 51 O.~
O

=~
~ N O O~
=' N ' N

[0531] [4-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-ethoxy-phenyl)-methanone 51 was prepared using the same protocol as described in Example 74, substituting 2-ethoxy naphthoyol chloride with 2-ethoxy-benzoyl chloride. MS(ESI) [M+H+]+ = 403.3.

Example 76: Synthesis of compounds of Formula Ic Scheme - 23 Br A A
v--~ ._- -u 1 x Stepl u ~~ x y Step2 u ~ x y y tN tN tN
H H =
k XIII XIV
Formula Ic Step -1 Synthesis offormula XIV
[0532] Compound XIII can be prepared from 7-azaindole following published procedure (Marie-Claude, Viaud, Heterocycles, 1999, 50, 1065-1080), and compound of Formula XIV
can be prepared from compound XIII by reacting with boronic acids under Suzuki reaction conditions (aqueous base and Pd(0) catalyst or anhydrous conditions with KF in dioxane with Pd(0) catalyst). Alternately, compound XIII can also be reacted with a tin reagent, zinc reagent or copper reagent, under Stille, Negishi or cuprate coupling reaction conditions respectively, to provide compound XIV. The product can be isolated by conventional work up procedure, e.g. extraction of the product with an organic solvent and purifying by column chromatography.

Step -2 Synthesis of Formula Ic [0533] Compound of Formula Ic can be prepared by reacting compound XIV witll nucleophilic reagents, e.g. acid chlorides, sulfonyl chlorides, isocyanates, isothiocyanates, alkyl halides, benzyl halides, etc., under basic conditions. The product can be isolated by following standard workup procedures, e.g. extraction of the product with organic solvent and purifying by coluinn chromatography.

Example 77: Synthesis of [5-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-ethoxy-naphthalen-l-yl)-methanone 80 Scheme - 24 O
'O 'O
.0O
Br ' Step 1 Step 2 N
N 0_ N
H N N p H
O".0 Step-1 Preparation of 5-(3, 4-Dimethoxy phenyl)-1 H-pyrrolo[2, 3-b]pyridine 82 [0534] In a microwave safe tube, 5-bromo-7-azaindole (81, 392 mg, 1.99 mmol), prepared from 7-azaindole following the published procedure (Marie-Claude, Viaud, Heterocycles, 1999, 50, 1065-1080), 3,4-dimethoxyphenyl boronic acid (905 mg, 4.97 mmol), and tetrakis(triphenylphosphine)palladium(0) (11 mg, 0. 099 mol) were mixed in 1.0 M of potassium carbonate (6.0 mL) and tetrahydrofuran (9.5 mL, 0.12 mol). The resulting mixture was heated at 120 C in a CEM Discover microwave unit for 10 minutes. Ethyl acetate and water were added, and the two layers were separated. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine and dried over anhydrous sodium sulfate. Concentration under reduced pressure afforded the crude material, which was purified by column chromatography (40 - 70% ethyl acetate: hexanes) to yield the desired product as a light yellow solid (82, 207 mg, 41%). MS(ESI) [M+H+]+ = 255.2.

Step-2 Preparation of [5-(3, 4-Dimethoxy phenyl) pyrrolo[2, 3-bJpyidin-1 ylJ-(2-ethoxy-naphthalen-1 yl)-methanone 80 [0535] Sodium hydride (60% dispersion in mineral oil, 10.1 mg, 0.252 mmol) was washed with hexane and put under an atmosphere of nitrogen, and 1 mL of tetrahydrofuran was added.
5-(3,4-Dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine (82, 49.3 mg, 0.194 mmol) in tetrahydrofuran (1.6 mL) was added, and the resulting mixture was stirred for 30 minutes at room temperature. 2-Ethoxy naphthoyol chloride (51.1 mg, 0. 213 mmol) in THF
was added.
After two hours, the reaction was quenched with water, and the two layers were seperated.
The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine and dried over anhydrous sodiu.ln sulfate. Concentration under reduced pressure afforde the crude, which was purified by column chromatography (40 - 55% ethyl acetate in hexane) to yield the desired product as a liquid (80, 61 mg, 70%). MS(ESI) [M+H+]+ = 453.2.
Example 78: Synthesis of [5-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-ethoxy-phenyl)-methanone 83 ~O
.'O

~~ =
N N
O
O", [0536] [5-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-ethoxy-phenyl)-methanone 83 was prepared using the same protocol as described in Example 77, substituting 2-ethoxy naphthoyol chloride with 2-ethoxy-benzoyl chloride. MS(ESI) [M+H+]+ = 403.2.

Example 79: Synthesis of (2-Ethoxy-naphthalen-1-yl)-[3-(4-methanesulfonyl-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-methanone 84 O.~
S
"O
/ =
~
N= ~ ~ ~
N _ ~ ~
O
O
/

Scheme - 25 O.
"O
~=
Br ~
' ' Br Step1 Ste-C N N ~ . s ~
N N ~ N N
H I ~
3 O I~ O
O
85 > 84 Step 1- Preparation of (3-BYomo pyrrolo[2, 3-b]pyridin-1 yl)-(2-ethoxy-naphthalen-l-yl)-methanone 85 [0537] 3-Bromo-7-azaindole (500 mg, 2.0 mmol) 3 was dissolved in N,N-dimethylformamide (50 mL) and sodium hydride (210 mg, 5.3 mmol, 60% dispersion in mineral oil) and 2-Ethoxy-naphthalene-l-carbonyl chloride (710 mg, 3.0 mmol) were added.
The reaction mixture was stirred at ambient temperature for 30 min, cast into ice water (100 mL) and extracted into ethyl acetate. The organic portion was dried with anhydrous magnesium sulfate, filtered and the filtrate concentrated. Purification via colurnn chromatography (10% Ethyl acetate in hexanes) provided the desired product 85 (800 mg, 80 %).

Step 2- Preparation of (2-Ethoxy-naphthalen-1 yl)-[3-(4-methanesulfonyl phenyl)-pyrrolo[2,3-b]pyridin-1 ylJ-methanone 84 [0538] (3-Bromo-pyrrolo[2,3-b]pyridin-l-y 1)-(2-ethoxy-naphthalen-1-yl)-methanone 85 (35 mg, 0.0088 mmol), 4-Methanesulfonyl-phenylboronic acid (35 mg, 0.18 mmol) and tetrakis(triphenylphosphine)palladium(0) (5 mg) were stirred in tetrahydrofuran (16 mL) and potassium carbonate solution (8.0 mL, 1M aqueous). The reaction mixture was stirred over night at 60 C. The reaction mixture was concentrated tulder reduced pressure and partitioned between ethyl acetate and brine. The organic portion was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated and purified by flash chromatography (ethyl acetate: hexanes 20%-100%). The desired product, 84 was obtained as a pale yellow powder (10 mg, 20 %).

Example 80: Synthesis of (2-Ethoxy-naphthalen-1-yl)-[3-(3-methanesulfonyl-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-methanone 86 / = S' O

N N
~
o ~s ~

[0539] (2-Ethoxy-naphthalen-1-yl)-[3-(3-methanesulfonyl-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-methanone 86 was prepared using the same protocol as described in Exainple 79, substituting 4-Methanesulfonyl-phenylboronic acid with 3-Methanesulfonyl-phenylboronic acid.

Example 81: Synthesis of 3-[1-(2-Ethoxy-naphthalene-l-carbonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzoic acid benzyl ester 87 O
N
O

~.~ x N N
O
O

[0540] 3-[1-(2-Ethoxy-naphthalene-l-carbonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzoic acid benzyl ester 87 was prepared using the same protocol as described in Example 79, substituting 4-Methanesulfonyl-phenylboronic acid with 3-carbobenzyloxy-phenylboronic acid.

Example 82: Synthesis of 3-(3-Cyclopentyloxy-4-methoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 88 /
O

O

N N
H

(0541] 3-(3-Cyclopentyloxy-4-methoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 88 was prepared using the same protocol as described in Example 7, substituting 3,4-dimethoxyphenylboronic acid with 3-cyclopentyloxy,4-methoxy-phenylboronic acid. MS(ESI) [M+H+]+=
309.20.

Example 83: Synthesis of 3-(3-Benzyloxy-4-methoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 89 /
O

/ = = /

N N
H

[0542] 3-(3-Benzyloxy-4-methoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 89 was prepared using the same protocol as described in Example 7, substituting 3,4-dimethoxyphenylboronic acid witli 3-benzyloxy,4-methoxy-phenylboronic acid.

Example 84: Synthesis of 3-Benzo[1,3]dioxol-5-yl-lH-pyrrolo[2,3-b]pyridine 90 O = /

N N
H
[0543] 3-Benzo[1,3]dioxol-5-yl-lH-pyrrolo[2,3-b]pyridine 90 was prepared using the same protocol as described in Example 7, substituting 3,4-dimethoxyphenylboronic acid with 3,4-methylenedioxybenzene boronic acid.

Example 85: Synthesis of 1-(Benzo[b]thiophene-3-sulfonyl)-3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 91 /
O

O = /
=~
N N
S~O

S

[0544] 1-(Benzo[b]thiophene-3-sulfonyl)-3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 91 was prepared using the same protocol as described in Example 9, substituting 8-quinoline-sulfonyl chloride with Benzo[b]thiophene-3-sulfonyl chloride.
MS(ES1) [M+H+]+=
450.97.

Example 86: Synthesis of 8-[3-(3,4-Dimethoxy-phenyl)-5-methoxy-indole-l-sulfonyl]-quinoline 92 /
O

0 Ol =
CbO 92 [0545] 8-[3-(3,4-Dimethoxy-phenyl)-5-methoxy-indole-l-sulfonyl]-quinoline 92 was prepared using the same protocol as described in Examples 21 and 22, substituting 1-benzenesulfonyl-3-bromoindole with 1-Benzenesulfonyl-3-bromo-5-methoxyindole.
MS(ESI) [M+H+]+= 475.10.

Example 87: Synthesis of 8-[3-(3,4-Dimethoxy-pheiryl)-5-chloro-indole-l-sulfonyl]-quinoline 93 /
O

=~
N N
~= 4-Op [0546] 8-[3-(3,4-Dimethoxy-phenyl)-5-chloro-indole-l-sulfonyl]-quinoline 93 was prepared using the same protocol as described in Examples 21 and 22, substituting 1-Benzenesulfonyl-3-bromoindole with 1-Benzenesulfonyl-3-bromo-5-chloroindole.
MS(ESI) [M+H+]+= 479.10.

Example 88: Syntliesis of 8-[3-(3,4-Dimethoxy-phenyl)-5-methoxy-indole-l-sulfonyl]-3-methyl-quinoline 94 /
O

0 Ol N N
p [0547] 8-[3-(3,4-Dimethoxy-phenyl)-5-methoxy-indole-l-sulfonyl]-3-methyl-quinoline 94 was prepared using the same protocol as described in Examples 21 and 22, substituting 8-quinoline-sulfonyl chloride with 3-methyl-quinoline-8-sulfonyl chloride.
MS(ESI) [M+H+]+=
489.10.

Example 89: Synthesis of 8-[5-Chloro-3-(3-cyclopentyloxy-4-methoxy-phenyl)-indole-l-sulfonyl]-3-methyl-quinoline 95 /
O

O CI
%-N N
S.-00 [0548] 8-[5-Chloro-3-(3-cyclopentyloxy-4-methoxy-phenyl)-indole-l-sulfonyl]-3-methyl-quinoline 95 was prepared using the same protocol as described in Examples 21 and 22, substituting 8-quinoline-sulfonyl chloride, 1-Benzenesulfonyl-3-bromoindole and 3,4-dimethoxyphenyl boronic acid with 3-methyl-quinoline-8-sulfonyl chloride, 1-Benzenesulfonyl-3-bromo-5-chloro-indole and 3-cyclopentyloxy-4-methoxy-phenylboronic acid respectively. MS(ESI) [M+H+]+= 547.10.

Example 90: Synthesis of 8-[5-Chloro-3-(3,4-Dimethoxy-phenyl)-indole-l-sulfonyl]-3-methyl-quinoline 96 /
O

N

[0549] 8-[5-Chloro-3-(3,4-Dimethoxy-phenyl)-indole- l-sulfonyl]-3-methyl-quinoline 96 was prepared using the same protocol as described in Examples 21 and 22, substituting 8-quinoline-sulfonyl chloride and 1-Benzenesulfonyl-3-bromoindole with 3-methyl-quinoline-8-sulfonyl chloride and 1-benzenesulfonyl-3-bromo-5-methoxy-indole respectively. MS(ESI) [M+H+]+=
493.00.

Example 91: Synthesis of 8-[5-Methoxy-3-(3-cyclopentyloxy-4-methoxy-phenyl)-indole-l-sulfonyl]-3-methyl-quinoline 97 /
O
O Ol d s~
%-N N

[0550] 8-[5-Methoxy-3-(3-cyclopentyloxy-4-methoxy-phenyl)-indole-l-sulfonyl]-3-methyl-quinoline 97 was prepared using the same protocol as described in Examples 21 and 22, substituting 8-quinoline-sulfonyl chloride, 1-Benzenesulfonyl-3-bromoindole and 3,4-dimethoxyphenyl boronic acid with 3-methyl-quinoline-8-sulfonyl chloride, 1-benzenesulfonyl-3-bromo-5-methoxy-indole and 3-cyclopentyloxy-4-methoxy-phenylboronic acid respectively. MS(ESI) [M+H+]+= 543.20.

Example 92: 3-(3-Cyclopentyloxy-4-methoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3-nitro-4-fluoro-phenyl)-amide 98 /
O

O = /

N N
H.N,IlzO
~ NO2 F

[0551] 3-(3-Cyclopentyloxy-4-methoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carboxylic acid (3-nitro-4-fluoro-phenyl)-amide 98 was prepared using the same protocol as described in Example 25, substituting 3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 1 and 4-chlorophenyl isocyanate with 3-(3-Cyclopentyloxy-4-methoxy-phenyl)-IH-pyrrolo[2,3-b]pyridine 88 and 4-fluoro-3-nitrophenyl isocyanate respectively. MS(ESI) [M+H+]}= 491.10.
Example 93: 1-Benzenesulfonyl-3-(3-cyclopentyloxy-4-methoxy-phenyl)-1H-indole /
O

O = /

N
0=8;0 ~I
~

[0552] 1-Benzenesulfonyl-3-(3-cyclopentyloxy-4-methoxy-phenyl)-1H-indole 99 was prepared using the same protocol as described in Example 21, substituting 3,4-dimethoxyphenyl boronic acid with 3-cyclopentyloxy-4-methoxy-phenyl boronic acid .
MS(ESI) [M+H+]+= 448.31.

Example 94: 8-[3-(3-Cyclopentyloxy-4-inethoxy-phenyl)-indole-l-sulfonyl]-quinoline 100 /
O
O = /

N
O=S:O
N
~ i [0553] 8-[3-(3-Cyclopentyloxy-4-methoxy-phenyl)-indole-l-sulfonyl]-quinoline 100 was prepared using the saine protocol as described in Examples 21 and 22, substituting 3,4-dimethoxyphenyl boronic acid and benxene sulfonyl chloride with 3-cyclopentyloxy-4-methoxy-phenyl boronic acid and 8-quinoline sulfonyl cliloride respectively.
MS(ESI) [M+H+] 499.09.

Example 95: 1-Benzenesulfonyl-3-(3-cyclopentyloxy-4-methoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 101 /
O

O = /

N N
O:S:O
~I
~

[0554] 1-Benzenesulfonyl-3-(3-cyclopentyloxy-4-methoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 101 was prepared using the same protocol as described in Example 7, substituting 3,4-dimethoxyphenyl boronic acid with 3-cyclopentyloxy-4-methoxy-phenyl boronic acid.
MS(ESI) [M+H' ]+= 449.13.

Example 96: 8-[3-(3-Cyclopentyloxy-4-methoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-sulfonyl]-quinoline 102 /
O

O

N N
O:S:O
~ ' N~
~ i [0555] 8-[3-(3-Cyclopentyloxy-4-methoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-sulfonyl]-quinoline 102 was prepared using the same protocol as described in Example 42, substituting 2-metlioxy-pyrimidine-4-boronic acid with 3-cyclopentyloxy-4-methoxy-phenyl boronic acid.
MS(ESI) [M+H+]+= 500.20.

Example 97: 1-Benzenesulfonyl-3-(3-benzyloxy-4-methoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 103 /
O

O
- ~~
N N
OO
~I
~

[0556] 1-Benzenesulfonyl-3-(3-benzyloxy-4-methoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 103 was prepared using the same protocol as described in Example 42, substituting 2-methoxy-pyrimidine-4-boronic acid and quinoline-8-sulfonyl chloride with 3-cyclopentyloxy-4-methoxy-phenyl boronic acid and benzene sulfonyl chloride respectively.

Example 98: 3-(3,4-Dimethoxy-phenyl)-1-(3-phenyl-isoxazol-5-ylmethyl)-lH-pyrrolo[2,3-b]pyridine 104 /
O
= -O

N N

ON =/

[0557] 3-(3,4-Dimethoxy-phenyl)-1-(3-phenyl-isoxazol-5-ylmethyl)-1H-pyrrolo[2,3-b]pyridine 104 was prepared using the same protocol as described in Example 46, substituting m-nitro-benzyl chloride with 5-Chloromethyl-3-phenyl-isoxazole. MS(ESI) [M+H+]+= 412.23.
Example 99: [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-[5-(3-iodo-phenyl)-isoxazol-3-yl]-methanone 105 /

= -O = /

N N
O ~= -N,O = /

[0558] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-[5-(3-iodo-phenyl)-isoxazol-3-yl]-methanone 105 was prepared using the same protocol as described in Example 45, substituting 2-ethoxy-l-naphthoyl chloride with 3-(3-Iodo-phenyl)-isoxazole-5-carbonyl chloride.

Example 100: 4-[3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carbonyl]-benzonitrile /

= -O
=~
N N
O ~~
~
CN

[0559] 4-[3-(3,4-Diinethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carbonyl]-benzonitrile 106 was prepared using the same protocol as described in Example 45, substituting 2-ethoxy-1-naphthoyl chloride with 4-cyano-benzoyl chloride. MS(ESI) [M+H+]+=384.20.

Example 101: (6-Chloro-pyridin-3-yl)-[3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-methanone 107 /
O
= -O

N N
O In N CI

[0560] (6-Chloro-pyridin-3-yl)-[3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-methanone 107 was prepared using the same protocol as described in Example 45, substituting 2-ethoxy-1-naphthoyl chloride with 2-chloro nicotinoyl chloride. MS(ESI) [M+H'-]+= 394.10.

Example 102: [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-((1R,2R)-2-phenyl-trans-cyclopropyl)-inethanone 108 /

= -O
=~
N
H N
O
IH

[0561] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-((1R,2R)-2-phenyl-trans-cyclopropyl)-methanone 108 was prepared using the same protocol as described in Example 45, substituting 2-ethoxy-l-naphthoyl chloride with tf=ans-2-phenylcyclopropane-carbonyl chloride. MS(ESI) [M+H+]+= 399.17.

Example 103: [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(4-dimethylamino-naphtlialen-1-yl)-methanone 109 /
O
= -O

N N
O

~~ -[0562] [3-(3,4-Diinethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(4-dimethylamino-naphthalen-l-yl)-methanone 109 was prepared using the same protocol as described in Example 45, substituting 2-ethoxy-l-naphthoyl chloride with 4-Dimethylamino-naphthalene-1-carbonyl chloride. MS(ESI) [M+H+]+= 452.20.

Example 104: Acetic acid 2-[3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carbonyl]-phenyl ester 110 /
O
= -O = /

N N
O
O
--~O

[0563] Acetic acid 2-[3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridine-l-carbonyl]-phenyl ester 110 was prepared using the same protocol as described in Example 45, substituting 2-ethoxy-l-naphthoyl chloride with Acetic acid 2-chlorocarbonyl-phenyl ester.
MS(ESI) [M+H+]+= 417.20.

Example 105: (2,4-Diethoxy-phenyl)-[3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-methanone 111 /
O
= -O = /
=~
N N

O iI j O
O

[0564] (2,4-Diethoxy-phenyl)-[3-(3,4-dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-methanone 111 was prepared using the same protocol as described in Example 45, substituting 2-ethoxy-l-naphthoyl chloride with 2,4-dimethoxy-benzoyl chloride. MS(ES1) [M+H+]+=
447.20.

Example 106: [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-[2-(2,2,2-trifluoro-ethoxy)-naphthalen-1-yl] -methanone 112 /

= -F 0 ~~ ~60 [0565] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-[2-(2,2,2-trifluoro-ethoxy)-naplithalen-l-yl]-methanone 112 was prepared using the same protocol as described in Example 47, substituting 2-ethoxy-4-nitro-benzoic acid with 2-(2,2,2-Trifluoro-ethoxy)-naphthalene- 1 -carboxylic acid. MS(ESI) [M+H+]+= 507.10.

Example 107: [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-(2-propoxy-phenyl)-methanone 113 /

= -O

I
N N
O
O

[0566] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-(2-propoxy-phenyl)-methanone 113 was prepared using the same protocol as described in Example 47, substituting 2-ethoxy-4-nitro-benzoic acid with 2-propoxy-naphthalene-l-carboxylic acid.
MS(ESI) [M+H+]+= 417.20.

Example 108: [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(4-methyl-naphthalen-l-yl)-methanone 114 /

= -O
/ ~ B
~

[0567] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(4-methyl-naphthalen-l-yl)-methanone 114 was prepared using the same protocol as described in Example 47, substituting 2-ethoxy-4-nitro-benzoic acid with 4-methyl-naphthalene-l-carboxylic acid.
MS(ESI) [M+H+]+= 423.20.

Example 109: [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-quinolin-4-yl-methanone 115 /

= -O = /

N N
O ~
N

[0568] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-quinolin-4-yl-methanone 115 was prepared using the same protocol as described in Example 47, substituting 2-ethoxy-4-nitro-benzoic acid with quinoline-4-carboxylic acid. MS(ESI) [M+H+]+= 410.20.

Example 110: [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-methyl-quinolin-4-yl)-methanone 116 /

= -O
N

[0569] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-methyl-quinolin-4-yl)-metllanone 116 was prepared using the same protocol as described in Example 47, substituting 2-ethoxy-4-nitro-benzoic acid with 2-methyl-quinoline-4-carboxylic acid.
MS(ES1) [M+H+]+_ 424.20.

Example 111: [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(3-phenyl-quinolin-4-yl)-inethanone 117 /

= -O

N N
O
O
N
=

[0570] [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(3-phenyl-quinolin-4-yl)-methanone 117 was prepared using the same protocol as described in Example 47, substituting 2-ethoxy-4-nitro-benzoic acid with 3-phenyl-quinoline-4-carboxylic acid.
MS(ESI) [M+H+]+_ 486.20.

Example 112: [3-(3,4-Dimethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-phenoxy-phenyl)-methanone 118 /

= -O

N N
O ~~
O ~
i~
~

[0571] [3-(3,4-Diinethoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-(2-phenoxy-phenyl)-methanone 118 was prepared using the same protocol as described in Example 47, substituting 2-ethoxy-4-nitro-benzoic acid with 2-phenoxy-benxoic acid. MS(ESI) [M+H+]}=
451.16.
Example 113: [3-(3-Cyclopentyloxy-4-methoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-(2-ethoxy-naphthalen-1-yl)-methanone 119 /
Sb N
O
O

[0572] [3-(3-Cyclopentyloxy-4-methoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-(2-ethoxy-naphthalen-1-yl)-methanone 119 was prepared using the same protocol as described in Example 79, substituting 4-Methanesulfonyl-phenylboronic acid with 3-cyclopentoxy-4-methoxy phenyl boronic acid. MS(ESI) [M+H+]+= 507.30.

Example 114: 3-[1-(2-Ethoxy-naphthalene-l-carbonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzoic acid methyl ester 120 O
=/
O

N N
~, O ~=
O ~
J

[0573] 3-[1-(2-Ethoxy-naphthalene-l-carbonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzoic acid methyl ester 120 was prepared using the same protocol as described in Example 79, substituting 4-Methanesulfonyl-phenylboronic acid witlz 3-methoxycarbonyl phenyl boronic acid. MS(ESI) [M+H+]+= 451.14.

Example 115: 3-[1-(2-Ethoxy-naphthalene-l-carbonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzamide 121 =/
O D
N V
O J

[0574] 3-[1-(2-Ethoxy-naphthalene-l-carbonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzamide 121 was prepared using the same protocol as described in Example 79, substituting 4-Methanesulfonyl-phenylboronic acid with 3-amido phenyl boronic acid. MS(ESI) [M+H+]+=
436.14.

Example 116: N-{3-[1-(2-Ethoxy-naphthalene-l-carbonyl)-lH-pyrrolo[2,3-b]pyridin-3-yl]-phenyl}-methanesulfonamide 122 OliN =/
-O

N N
~
' O ~~
O ~
J

[0575] N-{3-[1-(2-Ethoxy-naphthalene-l-carbonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-phenyl}-methanesulfonamide 122 was prepared using the same protocol as described in Example 79, substituting 4-Methanesulfonyl-phenylboronic acid with 3-N-methanesulfonamide phenyl boronic acid. MS(ESI) [M+H+]+= 486.10.

Example 117: (2-Ethoxy-naphthalen-1-yl)-[3-(4-hydroxy-3-methoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-methanone 123 HO
= -O = /

N ~ O

[0576] (2-Ethoxy-naphthalen-1-yl)-[3-(4-hydroxy-3-methoxy-phenyl)-pyrrolo[2,3-b]pyridin-1-yl]-methanone 123 was prepared using the same protocol as described in Example 79, substituting 4-Methanesulfonyl-phenylboronic acid with 2-Methoxy-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenol. MS(ESI) [M+H+]+= 439.17.

Example 118: [3-(3-Cyclopentyloxy-4-methoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-[2-(2,2,2-trifluoro-ethoxy)-naphthalen-l-yl] -methanone 124 /
O

_ O ~ ~ ~
O ~
F~1' F

[0577] [3-(3-Cyclopentyloxy-4-methoxy-phenyl)-pyrrolo[2,3-b]pyridin-l-yl]-[2-(2,2,2-trifluoro-ethoxy)-naphthalen-1-yl]-methanone 124 was prepared using the same protocol as described in Example 47, substituting 2-ethoxy-4-nitro-benzoic acid and 3-(3,4-Dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine with 2-(2,2,2-Trifluoro-ethoxy)-naphthalene-l-carboxylic acid and 3-(3-cyclopentoxy-4-methoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine.
MS(ESI) [M+H+]+= 561.10.

Example 119: 1-Benzenesulfonyl-3-[3-methoxy-4-(3-phenyl-isoxazol-5-ylmethoxy)-phenyl]-1H-pyrrolo[2,3-b]pyridine 125 =
N O

õ ~O = /
=~
N N
O: S=O
=~

[0578] 1-Benzenesulfonyl-3-[3-methoxy-4-(3-phenyl-isoxazol-5-ylmethoxy)-phenyl]-1H-pyrrolo[2,3-b]pyridine 125 was prepared using the same protocol as described in Example 7, substituting 2,3-diinethoxy-phenyl boronic acid with 5-[2-Methoxy-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenoxymethyl]-3-phenyl-isoxazole. MS(ESI) [M+H+]+=
538.06.
Example 120: 1-Benzenesulfonyl-3-[3-methoxy-4-(3-phenyl-isoxazol-5-ylmethoxy)-phenyl]-1H-pyrrolo[2,3-b]pyridine 126 Scheme - 26 O/ O( ,o S02Ci ,O
i~ .

N N N N
H O:S=O
50 N~
i Preparation of 8-[4-(3,4-Dimethoxy phenyl) pyrrolo[2,3-bJpyf-idine-l-sulfonylJ-quinoline 126 [0579] Into a Round bottom flask was added 4-(3,4-Dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine (0.222 g, 0.000873 mol), 50 and Tetra-N-butylammonium bromide (0.0282 g, 0.0000874 mol), and 5.000 M of Sodium hydroxide in Water (2.25 mL). 8-quinoline-sulfonyl chloride (0.238 g, 0.00105 mol) dissolved in Methylene chloride (0.616 mL, 0.00960 mol) was added dropwise at 0 Celsius. The reaction was stirred at ambient temperature for 3 h and the reaction mixture was diliuted with au addional 25 mL of methylene chloride.
The organic layer was washed with 1M sodium bicarbonate (aq.) (30 ml x2) and then with brine. The separated organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (30 % ethyl acetate in hexanes) to yield the titled compound as a white solid. MS(ESI) [M+H+]+ = 446.20.

Example 121: 4-(3,4-Dimethoxy-phenyl)-1-phenylmethanesulfonyl-lH-pyrrolo[2,3-b]pyridine 127 O/
e0 N N
O: S=O
C

[0580] 4-(3,4-Dimethoxy-phenyl)-1-phenylmethanesulfonyl-lH-pyrrolo[2,3-b]pyridine 127 was prepared using the same protocol as described in Example 120, substituting 8-quinoline-sulfonyl chloride with benzyl sulfonyl chloride. MS(ESI) [M+H+]+= 409.20.

Example 122: 4-(3,4-Dimethoxy-phenyl)-1H-indole 128 Scheme - 27 O~
~ O~
O 0 Br (HO)2B~

/ b i N N

Syntlaesis of 4-(3,4-Dinaethoxy phenyl)-1H-indole 128 [0581] In a microwave safe tube, 4-Bromoindole (1.383 g, 0.007054 mol), 3,4-dimethoxyphenyl boronic acid (3.21 g, 0.0176 mol), and Tetrakis(triphenylphosphine)palladium(0) (0.41 g, 0.00035 mol) were mixed in 1.00 M of Potassium carbonate in Water (21 mL) and Tetrahydrofuran (34 mL, 0.42 mol).
The resulting mixture was heated at 100 Celsius in the microwave for 10 minutes. The reaction mixture was partitioned between water and ethyl acetate. The aqueous layer was extracted with ethyl acetate and the organic layers were combined, washed with brine and dried over sodium sulfate. The reaction mixture was concentrated under reduced pressure and the residue was purified by silica gel chromatography (30 % ethyl acetate in hexanes) to the yield 1.02 g of the titled compound as a light green solid. MS(ESI) [M+H]+ = 254.20.

Example 123: 5-(3,4-Dimethoxy-phenyl)-1H-indole 129 O
Ol =~
=~

N
H

[0582] 5-(3,4-Dimethoxy-phenyl)-1H-indole 129 was prepared using the same protocol as described in Example 122, substituting 4-bromoindole with 5-bromoindole.
MS(ESI) [M+H+]+= 254.20 Example 124: 8-[5-(3,4-Dimethoxy-phenyl)-indole-l-sulfonyl]-quinoline 130 Ol =~
=

N
O:S:O

= 130 [0583] 8-[5-(3,4-Dimethoxy-phenyl)-indole-l-sulfonyl]-quinoline 130 was prepared using the same protocol as described in Example 9, substituting 3-(3,4-Dimethoxy-phenyl)-1H-pyrrolo-[2,3-b]pyridine with 5-(3,4-Dimethoxy-phenyl)-1H-indole. MS(ESI) [M+H+]+= 445.20 Example 125: 8-[4-(3,4-Dimethoxy-phenyl)-indole-l-sulfonyl]-quinoline 131 O
s0 =
=~
N
O=S:O
60;

[0584] 8-[4-(3,4-Dimethoxy-phenyl)-indole-l-sulfonyl]-quinoline 131 was prepared using the same protocol as described in Example 9, substituting 3-(3,4-Dimethoxy-phenyl)-1H-pyrrolo-[2,3-b]pyridine with 4-(3,4-Dimethoxy-phenyl)-1H-indole. MS(ESI) [M+H+]+= 445.10 Example 126: 1-Benzenesulfonyl-5-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 132 O( I
=~

N N
O
=~
~

[0585] 1-Benzenesulfonyl-5-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine 132 was prepared using the same protocol as described in Example 9, substituting 3-(3,4-Dimethoxy-phenyl)-1H-pyrrolo-[2,3-b]pyridine and quioline-8-sulfonyl chloride with 5-(3,4-Dimethoxy-phenyl)-1H-pyrrolo-[2,3-b]pyridine and benzene sulfonyl chloride respectively.
MS(ESI) [M+H+]+= 395.20 Example 127: 4-[I-Benzenesulfonyl-5-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-ylethynyl]-benzoic acid ethyl ester 135 /
O~
\ ~ \
~
N N
0=S=O
a Scheme - 28 O O' O

Step 1 I Step2 N N N N

H H

O
~ ( - ~
O O~ \ \ ~ O~
Step 3 O

N N N N
0=S=0 0F 134 , I
~
Step 1-Pf eparation of 5-(3,4-Dimethoxyphenyl)-3-iodo-1Hpyrrolo[2,3-bJpyridine [0586] 5-(3,4-Dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine, 82, (0.270 g, 0.00106 mol) was dissolved in Tetrahydrofitran (8.5 mL, 0.10 mol) under an atmosphere of Nitrogen. The solution was stirred at -40 C and Iodine (0.269 g, 0.00106 mol) dissolved in 2.5 mL of Tetrahydrofuran was added. The chilled reaction mixture was stirred for 2 h and was then quenched with the addition of Sodium thiosulfate, pentahydrate (0.13 g, 0.00053 mol) in water (1M). The reaction mixture was partitioned between water (20 inL and ethyl acetate (30 mL). The two layers were seperated, and the aquous layer was extracted with ethyl acetate.
The organic layers were washed with water and brine, dried with sodium sulfate, then concentrated under reduced pressure. The dark colored crude residue was carried onto the next reaction without further purification.

Step 2- Preparation of 1-Benzenesulfonyl-5-(3,4-dimethoxy phenyl)-3-iodo-IH-pyrrolo[2,3-bJpyridine 134 [0587] Into a Round bottom flask was added 5-(3,4-Dimethoxy-phenyl)-3-iodo-lH-pyrrolo[2,3-b]pyridine,133, (0.403 g, 0.00106 mol) and Tetra-N-butylammonium bromide (0.0342 g, 0.000106 mol), in 5.000 M of Sodium liydroxide in Water (2.73 mL).
Benzenesulfonyl chloride (0.225 g, 0.00127 mol) in Methylene chloride (0.747 mL, 0.0116 mol) was added dropwise. After 2 h, 30 mL of Methylene chloride and 30 mL of water were added. The organic layer was separated and washed with 1M sodium bicarbonate (aq.) (30 ml x2) followed by water (30 ml) and brine (30 mL). The organic layer was collected and dried over anhydrous sodium sulfate, then concentrated under reduced pressure. The residue was purified by chromatography (Silica gel, ethyl acetate/hexanes) to give 295 mg of the desired product as a white solid. MS(ESI) [M+H+]+= 521.04 Step 3-Prepaz~ation of 4-[1-Benzenesulfonyl-5-(3,4-dimethoxy phenyl)-1H-pyrrolo[2,3-bJpyridin-3 ylethynylJ-benzoic acid ethyl ester 135 [0588] 1-Benzenesulfonyl-5-(3,4-dimethoxy-phenyl)-3-iodo-lH-pyrrolo[2,3-b]pyridine (0.0800 g, 0.000151 mol), 4-Ethynyl-benzoic acid ethyl ester (0.0321 g, 0.000181 mol), Bis(triphenylphosphine)palladium(II) chloride (0.0048 g, 0.0000069 mol), and Copper(I) iodide (0.00024 g, 0.0000013 mol) were dissolved in Triethylamine (0.8 mL, 0.005 mol) under an atmosphere of Nitrogen. The resulting mixture was heated to 60 C and stirred under an atmosphere of Nitrogen for 2 hours. The reaction mixture was was concentrated under reduced pressure and water (30 mL) was added to the residue. This slurry was extracted with ether.
(20 mL 2X). The combined organic layers were washed with brine, dried over sodium sulfate and concentrated under reduced pressure. The crude was purified by chromatography (Silica gel, ethyl acetate/hexanes to give 85 mg of the titled product as a pale orange solid. MS(ESI) [M+H+]+= 567.10 Example 128: 1-Benzenesulfonyl-5-(3,4-dimethoxy-phenyl)-3-phenylethynyl-lH-pyrrolo[2,3-b]pyridine 136 O
O
\\ ~

N N
0=S=0 I

[0589] 1-Benzenesulfonyl-5-(3,4-dimethoxy-phenyl)-3-phenylethynyl-lH-pyrrolo[2,3-b]pyridine 136 was prepared using the same protocol as described in Example 127, substituting 4-Ethynyl-benzoic acid ethyl ester with Ethynyl-benzene. MS(ESI) [M+H+]+=
495.20 Example 129: 3-[1-Benzenesulfonyl-5-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-ylethynyl]-benzoic acid methyl ester 137 O \ / p O' 0 \\ ~ \

N N
0=~--0 I

[0590] 3-[1-Benzenesulfonyl-5-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-ylethynyl]-benzoic acid methyl ester 137 was prepared using the same protocol as described in Example 127, substituting 4-Ethynyl-benzoic acid ethyl ester with 3-Ethynyl-benzoic acid methy ester. MS(ESI) [M+H+]+= 553.10 Example 130: 5-(3,4-Dimethoxy-phenyl)-3-phenylethynyl-lH-pyrrolo[2,3-b]pyridine 138 O
O
\\ ~

N N
H

[0591] 5-(3,4-Dimethoxy-phenyl)-3-phenylethynyl-lH-pyrrolo[2,3-b]pyridine 138 was prepared using the same protocol as described in Example 7, substituting 1-Benzenesulfonyl-3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine with 1-Benzenesulfonyl-5-(3,4-dimethoxy-phenyl)-3-phenylethynyl-lH-pyrrolo[2,3-b]pyridine. MS(ESI) [M+H}]}=
355.20 Example 131: 3-[5-(3,4-Diinethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-ylethynyl]-benzoic acid methyl ester 139 O \ / O O\
0 \\ ~ \

N N
H

[0592] 3-[5-(3,4-Dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-ylethynyl]-benzoic acid methyl ester 139 was prepared using the same protocol as described in Example 7, substituting 1-Benzenesulfonyl-3-(3,4-dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine with 3-[1-B enzenesulfonyl-5 -(3,4-dimethoxy-phenyl)-1 H-pyrrolo [2, 3-b]pyridin-3 -ylethynyl] -b enzoic acid methyl ester. MS(ESI) [M+H+]+= 413.20 Example 132: 3-[5-(3,4-Dimethoxy-phenyl)-1 H-pyrrolo [2,3-b]pyridin-3-ylethynyl]-benzoic acid 140 Scheme - 29 O O O O O
O HO Ol N N N N
H H

Preparation of 3-[5-(3,4-Difmthoxy phen.yl)-IH pyrrolo[2,3-b]pyridin-3 ylethynylJ-benzoic acid 140 [0593] Into a microwave safe tube, 3-[5-(3,4-Dimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-ylethynyl]-benzoic acid methyl ester (0.049 g, 0.00012 mol) was dissolved in 5.00 M of Sodium hydroxide in Water (1.78 mL) and Methanol (7.1 mL, 0.18 mol). The reaction was warmed at 60 watts to 100 C for 10 minutes. The resulting clear yellow solution was concentrated under reduced pressure. The resulting aqueous slurry was acidified to pH 5 with 1M HCl (aq) and was extracted into etl7yl acetate. The organic layer was washed with brine, dried over anhydrous sodium sulfate, then concentrated under reduced pressure.
The resulting residue was recrystalized from ethyl acetate and methanol to yield the titled compound as a white solid. MS(ESI) [M+H+]}= 399.20 Example 133: 5-(1-Benzenesulfonyl-lH-pyrrolo[2,3-b]pyridin-3-yl)-2-methoxy-phenol 141 Scheme - 30 O
/= - -- O =/ HO =/

N N N N
O=S=O 0=S=0 Preparation of 5-(1-Benzenesulfonyl-1 H pyrrolo[2, 3-b]pyridin-3 yl)-2-methoxy phenol 141 [0594] Into a Parr pressure reactor 1-Benzenesulfonyl-3-(3-benzyloxy-4-methoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine (92 mg, 0.00020 mol) was placed with Palladium (50 mg, 0.00005 mol) 10%, tetrahydrofuran (15 mL, 0.18 mol) and Methanol (1 mL, 0.02 mol) and HCl solution (3 mL, 0.04 mol). The reaction was shaken under an atinosphere of hydrogen at 50 psi for 4 hours. The mixture reaction was filtered through CeliteTM and concentrated under reduced pressure. The residue was purified by preparative TLC (ethyl acetate/hexanes 1:1) to give 31 mg of the titled product. MS(ESI) [M+H+]+ = 381.11.

Example 134: 3-Benzo[1,3]dioxol-5-yl-2-methyl-lH-pyrrolo[2,3-b]pyridine142 O'O
=/
N N
H

[0595] 3-Benzo[1,3]dioxol-5-yl-2-methyl-lH-pyrrolo[2,3-b]pyridine 142 was prepared using the same protocol as described in Example 8, substituting 3-Bromo-lH-pyrrolo[2,3-b]pyridine and 3,4-dimethoxy-phenyl boronic acid with 3-Bromo-2-methyl-lH-pyrrolo[2,3-b]pyridine and 5-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzo[1,3]dioxole respectively.
Example 135: 3-(3,4-Dimethoxy-phenyl)-2-methyl-lH-pyrrolo[2,3-b]pyridine 143 O O-=/
N N
H

[0596] 3-(3,4-Dimethoxy-phenyl)-2-methyl-lH-pyrrolo[2,3-b]pyridine 143 was prepared using the same protocol as described in Example 8, substituting 3-Bromo-lH-pyrrolo[2,3-b]pyridine with 3-Bromo-2-methyl-lH-pyrrolo[2,3-b]pyridine.

Example 136: (4-Amino-2-ethoxy-phenyl)-[3-(3,4-dimethoxy-phenyl)-pyrrolo [2,3-b]pyridin-1-yl] -methanone 144 i = -N N
O
0 NHa [0597] [3-(3,4-diunethoxy-phenyl)-pyrrolo[2,3-b]pyridine-1-yl] -(2-ethoxy-4-nitro-phenyl)-methanone, 54, (100mg, 0.22 mmol) was dissolved in EtOAc (30mL). A catalytic amount of 10%Pd/C (7mg) was added and the flask capped with rubber septa. The flask was evacuated and back filled with hydrogen twice. Finally, the reaction was stirred overnight under a hydrogen gas atmosphere (balloon). The reaction mixture was filtered over Celite , rinsed generously with EtOAc (2x75mL) and concentrated under reduced pressure to give the titled compound. (56mg, 60%) MS(ESI) [M+H+]+ = 418.24 Example 137: N- {4-[3-(3,4-Dimethoxy-phenyl)-pyrrolo [2,3-b]pyridine-l-carbonyl]-3-ethoxy-phenyl}-methanesulfonamide 145 /

= -O

N N

g~
O N' 11 H O

[0598] Into a round bottom flask was (4-amino-2-ethoxyphenyl)[3-(3,4-dimethoxyphenyl)-pyrrolo[2,3-b]pyridin-1-yl] methonone (56 mg, 0.00013 mol) in tetrahydrofuran (15 mL).
Sodium hydride (4.8 mg, 0.00020 mol, 60% dispersion in mineral oil) was added and Methanesulfonyl chloride (38 mg, 0.00034 mol) was added to reaction mixture, which was then stirred at room temperature over night. Top of FormColumn chromatoghraphy purification (silica gel) gave the titled compoundBottom of Form. MS(ESI) [M+H+]+= 496.08 Example 138: Cloning of PDE4B Phosphodiesterase Domain [0599] PDE4B cDNA sequence was ainplified from a Human Brain, hippocampus QUICK-Clone cDNA library (Clontech, #7169-1) by PCR using the following primers:

PDE4B-S: 5'-CCGAATT CATATG AGCATCTCACGCTTTGGAGTC-3' 34 mer PDE4B-A: 5'-TGTGCT CTCGAG TTA GCTGTGTCCCTCTCCCTCC-3' 34 mer [0600] An internal Ndel site was then engineered out by site directed mutagenesis using the following primers:

PDE4B-NDE1: 5'-GATATGTCTAAAC-~,bATGAGCCTGCTGGC-3' 29 mer PDE4B-NDE2: 5'-GCCAGCAGGCTCAI':,GTGTTTAGACATATC-3' 29 mer [0601] The resulting PCR fragment was digested with Ndel and SalI and subcloned into the pET 15 S vector.

[0602] In this expression plasmid, residues 152-528 of PDE4B (NCBI sequence JC1519, SEQ ID NO: 1) are in frame with an N-terminal His-tag followed by a thrombin cleavage site.
[0603] The sequence of pET15S, with multi-cloning site is shown below:

T7 promoter AGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCC
RBS
TCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACC

Ndel ATGGGCAGCAGCCATCATCATCATCATCACAGCAGCGGCCTGGTGCCGCGCGGCAGCCATATGGGATCCGG
M G S S H H H H H H S S G L V P R G S H M--------StuI SalI
AATTCAAAGGCCTACGTCGACTAGAGCCTGCAGTCTCGACCATCATCATCATCATCATTAATAAAAGGGCG
----------------------- *

Spel BamH2 _iGGCCGTTACTAGTGGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGG

IVEX-3 Primer Bpu1102 I T7 terminator CTGCTGCCACC"'.' ..ACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG
3'-PET Primer [0604] pET15S vector is derived from pET15b vector (Novagen) for bacterial expression to produce the proteins with N-terminal His6. This vector was modified by replacement of NdeI-BamHI fragment to others to create a SalI site and stop codon (TAG). Vector size is 5814 bp.
Insertion can be performed using Ndel-Sall site. The amino acid and nucleic acid sequences for the PDE4B phosphodiesterase domain utilized are provided in Table 3.

Example 139: Purification of PDE4B

[0605] PDE4B is purified from E. coli cells [BL21(DE3)Codon Plus(RIL) (Novagen)] grown in Terrific broth that has been supplemented with 0.2mM Zinc Acetate and 1mM
MgC12 and induced for 16-20h with 1 mM IPTG at 22 C. The centrifuged bacterial pellet (typically 200-250g from 16 L) is suspended in lysis buffer (0.1M potassium phosphate buffer, pH 8.0, 10%
glycerol, 1 mM PMSF). 100ug/ml of lysozyme is added to the lysate and the cells are lysed in a Cell Disruptor (MicroFluidics). The cell extract is clarified at 5000 rpm in a Sorvall SA6000 rotor for lh, and the supernatant is recentrifuged for another hour at 17000 rpm in a Sorvall SA
600 rotor. 5 mM imidazole (pH 8.0) is added to the clarified supematant and 2 ml of cobalt beads (50% slurry) is added to each 35 ml of extract. The beads are mixed at 4 C for 3-4 h on a Nutator and the beads are recovered by centrifugation at 4000 rpm for 3 min.
The pelleted beads are washed several times with lysis buffer and the beads are packed on a BioRad disposable colurnn. The bound protein is eluted with 3-4 column volumes of 0.1M imidazole followed by 0.25M imidazole, both prepared in lysis buffer. The protein eluted from the cobalt beads is concentrated on Centriprep-10 membranes (Amicon) and separated on a Pharmacia Superdex 200 column (26/60) in low salt buffer (25 mM Tris-HC1, pH 8.0, 150 mM
NaCl, 14 mM beta-mercaptoethanol). At this stage the PDE proteins are treated with thrombin for 16-20 hours at room temperature. The PDE proteins are further purified by anion exchange chromatography on a Pharmacia Source Q column (10/10) in 20 mM Tris-HCl pH 8 and 14 mM beta-mercaptoethanol using a NaCl gradient in an AKTA-FPLC (Phazmacia).

Example 140: Crystallization of PDE4B Phosphodiesterase Domain [0606] Crystals of PDE4B were grown in 30% PEG 400, 0.2M MgC12, 0.1M Tris pH
8.5, 1 mM binding ligand, 15.9 mg/ml protein at 4 C, using an Intelliplate (Robbins Scientific, Hampton) by mixing one microliter of protein with one microliter of precipitant. Data was collected to 1.4A.

[0607] Additionally, PDE4B crystals were grown in 20% PEG 3000, 0.2M Ca(OAc)2, 0.1M
Tris pH 7.0, 1 mM binding ligand, 15.9 mg/ml protein at 4 C, using an Intelliplate (Robbins Scientific, Hampton) by mixing one microliter of protein with one microliter of precipitant.
Data was collected to 1.7A.

Example 141: Structure Determination of PDE4B

[0608] The structure of PDE4B was solved using molecular replacement, using the previously deposited coordinates for PDE4B. The atomic coordinates for the PDE4B structure determined are provided in Table 1(coordinates for a co-ciystal structure is provided in Table 2).

Example 142: PDE Binding Assays [0609] Binding assays can be performed in a variety of ways, including a variety of ways known in the art. For example, as indicated above, binding assays can be performed using fluorescence resonance energy transfer (FRET) format, or using an AlphaScreen [0610] Alternatively, any method which can measure binding of a ligand to the cAMP-binding site can be used. For example, a fluorescent ligand can be used. When bound to PDE4B, the emitted fluorescence is polarized. Once displaced by inhibitor binding, the polarization decreases.

[0611] Determination of IC50 for compounds by competitive binding assays.
(Note that KI is the dissociation constant for inhibitor binding; KD is the dissociation constant for substrate binding.) For this system, the IC50, inhibitor binding constant and substrate binding constant can be interrelated according to the following formula:

[0612] When using radiolabeled substrate, KI = IC5o , and 1+ [L*]/KD
the IC50 - KI when there is a small amount of labeled substrate.
Example 143: PDE Activity Assay [0613] As an exemplary phosphodiesterase assay, the effect of potential modulators phosphodiesterase activity of PDE4B, PDE5A, and other PDEs was measured in the following assay format:

Renents [0614] Assay Buffer 50 mM Tris, 7.5 8.3 mM MgC1Z
1.7 mM EGTA
0.01% BSA
Store @ 4 degrees [0615] RNA binding YSi SPA beads Beads are 100 mg/ml in water. Dilute to 5 mg/ml in 18 mM Zn using 1M
ZnAcetate/ZnS04 solution(3:1) and water. Store @ 4 degrees.

[0616] Low control compounds Concentration of 20X DMSO Stock PDE1B: 8-methoxymethyl IBMX 20 mM
PDE2A: EHNA 10 mM
PDE3B: Milrinone 2 mM
PDE4D: Rolipram 10 mM
PDE5A: Zaprinast 10 mM
PDE7B: IBMX 40 mM
PDE10A: Dipyridamole 4 mM

[0617] Enzyme concentrations (2X final concentration. Diluted in assay buffer) PDE1B 50 ng/ml PDE2A 50 ng/ml PDE3B 10 ng/ml PDE4D 5 ng/ml PDE5A 20 ng/ml PDE7B 25 ng/ml PDE10A 5 ng/ml) [0618] Radioligands [3H] cAMP (Amersham TRK559). Dilute 2000X in assay buffer.
[3H] cGMP (Amersham TRK392). For PDE5A assay only. Dilute 2000X in assay buffer.

[0619] Protocol - Make assay plates from 2mM, 96 well master plates by transferring lul of - compound to 384 well plate using BiomekFx. Final concentration of compounds will be -100 M. Duplicate assay plates are prepared from each master plate so that compounds are assayed in duplicate.
- To column 23 of the assay plate add lul of 20X DMSO stock of appropriate control compound. These will be the low controls.
- Columns 1 and 2 of Chembridge library assay plates and columns 21 and 22 of the Maybridge library assay plates have lul DMSO. These are the high controls.
- Using BiomekFx, pipet 10 l of radioligand into each assay well, then, using the same tips, pipet 10 l of enzyme into each well.
- Seal assay plate with transparent cover. Centrifuge briefly @ 1000 RPM, them mix on plate shaker for 10 s.
- Incubate @ 30 for 30 min.
- Using BiomekFx, add 10 1 of bead mixture to each assay well. Mix beads thoroughly in reservoir immediately prior to each assay plate addition.
- Re-seal plate with fresh transparent cover. Mix on plate shaker for 10 s, then centrifuge for 1 min. @ 1000 RPM.
- Place plates in counting racks. Let stand for> 30 min, then count on Wallac TriLux using program 8.

- Analyze data as % inhibition of enzyme activity. Average of high controls =
0%
inhibition. Average of low controls = 100% inhibition.

Example 144: Expression and purification of PPARs for use in biochemical and cell assays Genetic engineering [0620] Plasmids encoding the human phosphodiesterases (PDEs) 4B and 4D were engineered using common polymerase chain reaction (PCR) methods. Both the full-length PDEs and truncated versions harboring just the PDE catalytic domains were engineered for heterologous expression. The relevant DNA sequences and encoded protein sequences used are shown for each (see below). The human PDE4B and PDE4D genes have several splice variants; the splice variants chosen for full-length expression are PDE4B2 (NCBI accession gi 292387) and PDE4D5 (NCBI accession gi 2735856). Complementary DNA cloned from various human tissues were purchased from Invitrogen, and these were used as substrates in the PCR reactions.
Specific custom synthetic oligonucleotide primers (Invitrogen, see below) were designed to initiate the PCR product, and also to provide the appropriate restriction enzyme cleavage sites for ligation with the plasmids.

[0621] The plasmid used for ligation with the catalytic domain-encoding PDE4B
and PDE4D
inserts was derived from pET15 (Novagen) for expression using E. coli. The plasmid used for ligation of the full-length PDE4B a.nd PDE4D inserts was pFastBacHT
(Invitrogen). In all of these cases the PDE was engineered to include a Histidine tag for purification using metal affinity chromatography.

Protein Expression and Purification of PDE4 Catalytic domains in E.coli:

[0622] For protein expression, plasmids containing genes of interest were transformed into E.coli strains BL21(DE3)RIL and transformants selected for on LB agar plates containing appropriate antibiotics. Single colonies were grown for 4hrs at 37 C in 200m1 LB media. For PDE4B and PDE4D all protein expression was performed by large scale fermentation using a 30L bioreactor. 400m1 of starter culture was added to 30L TB culture and allowed to grow at 37 C until an OD600nm of 2-5 was obtained. The culture was cooled to 20 C
and 0.5mM
IPTG, 1mM MgC12 and 0.2mM ZnOAc added, the culture was allowed to grow for a further 18hrs.

[0623] For protein purificatio all operations were carried out at 4 C. Frozen E.coli cell pellets were resuspended in lysis buffer and lysed using standard mechanical methods. Soluble proteins were purified via poly-Histidine tags using immobilized metal affinity purification (IMAC). For each of the PDE's purification was achieved using a 3 step purification process utilizing; IMAC, size exclusion chromatography and ion exchange chromatography. For both PDE4B and PDE4D, the poly-Histidine tag was removed using Thrombin (Calbiochem) before the final purification step.

[0624] For proteins provided for assay purposes, the above described expression conditions were carried out except purification was only 2-steps and the poly-histidine tag was not removed. Enzymes were stored in 50% glycerol.

Protein Expression and Purification of full length PDE4 isoforms in insect cells using Baculovirus PDE's expressed using standard protocols.

[0625] PDE4B2: The full-length human PDE4B2 isozyme with an N-terminal His6 tag and TEV cleavage site expressed in baculovirus infected insect cells. The enzyme was not purified from the cell lysates, so enzyme concentrations were not determined. Enzyme was stored in 50% glycerol at -20 .

[0626] PDE4D5: The full-length human PDE4D5 isozyme with an N-terminal His6 tag and TEV cleavage site expressed in baculovirus infected insect cells. The enzyme was not purified from the cell lysates, so enzyme concentrations were not determined. Enzyme was stored in 50% glycerol at -20 .

Plasmid sequence and PCR primer infortnation:
[0627] PDE4B:
P457. pET15S PDE4B S152-S528-X
taatacgactcactataggggaattgt gagcggataacaattcccctctagaaataattttgtttaactttaagaaggagatatacc atgggcagcagccatcatcatcatcatcacagcagcggcctggtgecgcgcggcagccat M G S S H H H H H H S S G L V P R G S H
atgagcatctcacgctttggagtcaacactgaaaatgaagatcacctggccaaggagctg M S I S R F G V N T E N E D H L A K E L
gaagacctgaacaaatggggtcttaacatctttaatgtggctggatattctcacaataga E D L N K W G L N I F N V A G Y S H N R
cccctaacatgcatcatgtatgctatattccaggaaagagacctcctaaagacattcaga P L T C I M Y A I F Q E R D L L K T F R
atctcatctgacacatttataacctacatgatgactttagaagaccattaccattctgac I S S D T F I T Y M M T L E D H Y H S D
gtggcatatcacaacagcctgcacgctgctgatgtagcccagtcgacccatgttctcctt V A Y H N S L H A A D V A Q S T H V L L
tctacaccagcattagacgctgtcttcacagatttggaaatcctggctgccatttttgca S T P A L D A V F T D L E I L A A I F A
gctgccatccatgacgttgatcatcctggagtctccaatcagtttctcatcaacacaaat A A I H D V D H P G V S N Q F L I N T N
tcagaacttgctttgatgtataatgatgaatctgtgttggaaaatcatcaccttgctgtg S E L A L M Y N D E S V L E N H H L A V
ggtttcaaactgctgcaagaagaacactgtgacatcttcatgaatctcaccaagaagcag G F K L L Q E E H C D I F M N L T K K Q
cgtcagacactcaggaagatggttattgacatggtgttagcaactgatatgtctaaacac R Q T L R K M V I D M V L A T D M S K H
atgagcctgctggcagacctgaagacaatggtagaaacgaagaaagttacaagttcaggc M S L L A D L K T M V E T K K V T S S G
gttcttctcctagacaactataccgatcgcattcaggtccttcgcaacatggtacactgt V L L L D N Y T D R I Q V L R N M V H C
gcagacctgagcaaccccaccaagtccttggaattgtatcggcaatggacagaccgcatc A D L S N P T K S L E L Y R Q W T D R I
atggaggaatttttccagcagggagacaaagagcgggagaggggaatggaaattagccca M E E F F Q Q G D K E R E R G M E I S P
atgtgtgataaacacacagcttctgtggaaaaatcccaggttggtttcatcgactacatt M C D K H T A S V E K S Q V G F I D Y I
gtccatccattgtgggagacatgggcagatttggtacagcctgatgctcaggacattctc V H P L W E T W A D L V Q P D A Q D I L
gataccttagaagataacaggaactggtatcagagcatgatacctcaaagtccctcacca D T L E D N R N W Y Q S M I P Q S P S P
ccactggacgagcagaacagggactgccagggtctgatggagaagtttcagtttgaactg P L D E Q N R D C Q G L M E K F Q F E L
actctcgatgaggaagattctgaaggacctgagaaggagggagagggacacagctaactc T L D E E D S E G P E K E G E G H S -gactagagcctgcagtctcgaccatcatcatcatcatcattaataaaagggcgaattcca gcacactggcggccgttactagtggatcc PCR Primers:

[0628] PDE4D:

P4437. pET15S PDE4D S316-V692-X
taatacgactcactataggggaattgt gagcggataacaattcccctctagaaataattttgtttaactttaagaaggagatatacc atgggcagcagccatcatcatcatcatcacagcagcggcctggtgccgcgcggcagccat M G S S H H H H H H S S G L V P R G S H
atgagtatcccaaggtttggagttaaaactgaacaagaagatgtccttgccaaggaacta M S I P R F G V K T E Q E D V L A K E L
gaagatgtgaacaaatggggtcttcatgttttcagaatagcagagttgtctggtaaccgg E D V N K W G L H V F R I A E L S G N R
cccttgactgttatcatgcacaccatttttcaggaacgggatttattaaaaacatttaaa P L T V I M H T I F Q E R D L L K T F K
attccagtagatactttaattacatatcttatgactctcgaagaccattaccatgctgat I P V D T L I T Y L M T L E D H Y H A D
gtggectatcacaacaatatccatgctgcagatgttgtccagtctactcatgtgctatta V A Y H N N I H A A D V V Q S T H V L L
tctacacctgctttggaggctgtgtttacagatttggagattcttgcagcaatttttgcc S T P A L E A V F T D L E I L A A I F A
agtgcaatacatgatgtagatcatcctggtgtgtccaatcaatttctgatcaatacaaac S A I H D V D H P G V S N Q F L I N T N
tctgaacttgccttgatgtacaatgattcctcagtcttagagaaccatcatttggctgtg S E L A L M Y N D S S V L E N H H L A V
ggctttaaattgcttcaggaagaaaactgtgacattttccagaatttgaccaaaaaacaa G F K L L Q E E N C D I F Q N L T K K Q
agacaatetttaaggaaaatggtcattgacatcgtacttgcaacagatatgtcaaaacac R Q S L R K M V I D I V L A T D M S K H
atgaatctactggctgatttgaagactatggttgaaactaagaaagtgacaagctctgga M N L L A D L K T M V E T K K V T S S G
gttcttcttcttgataattattccgataggattcaggttcttcagaatatggtgcactgt V L L L D N Y S D R I Q V L Q N M V H C
gcagatctgagcaacccaacaaagcctctccagctgtaccgccagtggacggaccggata A D L S N P T K P L Q L Y R Q W T D R I
atggaggagttcttccgccaaggagaccgagagagggaacgtggcatggagataagcccc M E E F F R Q G D R E R E R G M E I S P
atgtgtgacaagcacaatgcttccgtggaaaaatcacaggtgggcttcatagactatatt M C D K H N A S V E K S Q V G F I D Y I

gttcatcccctctgggagacatgggcagacctcgtccacectgacgcccaggatattttg V H P L W E T W A D L V H P D A Q D I L
gacactttggaggacaatcgtgaatggtaccagagcacaatcectcagagcccctctcct D T L E D N R E W Y Q S T I P Q S P S P
gcacctgatgacecagaggagggccggcagggtcaaactgagaaattccagtttgaacta A P D D P E E G R Q G Q T E K F Q F E L
actttagaggaagatggtgagtcagacacggaaaaggacagtggcagtcaagtgtaagtc T L E E D G E S D T E K D S G S Q V -gactagagcctgcagtctcgaccatcatcatcatcatcattaataaaagggcgaattcca gcacactggcggccgttactagtggatcc PCR Primers:

[0629] PDE4B2:

P4477. pFastBac PDE4B2 Ml-T564-X
tattccggattattcataccgtcccaccatcgggcgcggatctcggtccgaaacc atgtcgtactaccatcaccatcaccatcacgattacgatatcccaacgaccgaaaacctg M S Y Y H H H H H H D Y D I P T T E N L
tattttcagggccatatgaaggagcacgggggcaccttcagtagcaccggaatcagcggt Y F Q G H M K E H G G T F S S T G I S G
ggtagcggtgactctgctatggacagcctgcagccgctccagcctaactacatgectgtg G S G D S A M D S L Q P L Q P N Y M P V
tgtttgtttgcagaagaatcttatcaaaaattagcaatggaaacgctggaggaattagac C L F A E E S Y Q K L A M E T L E E L D
tggtgtttagaccagctagagaccatacagacctaccggtctgtcagtgagatggcttct W C L D Q L E T I Q T Y R S V S E M A S
aacaagttcaaaagaatgctgaaccgggagctgacacacctctcagagatgagccgatca N K F K R M L N R E L T H L S E M S R S
gggaaccaggtgtctgaatacatttcaaatactttcttagacaagcagaatgatgtggag G N Q V S E Y I S N T F L D K Q N D V E
atcccatctcctacccagaaagacagggagaaaaagaaaaagcagcagctcatgacccag I P S P T Q K D R E K K K K Q Q L M T Q
ataagtggagtgaagaaattaatgcatagttcaagcctaaacaatacaagcatctcacgc I S G V K K L M H S S S L N N T S I S R
tttggagtcaacactgaaaatgaagatcacctggccaaggagctggaagacctgaacaaa F G V N T E N E D H L A K E L E D L N K
tggggtcttaacatctttaatgtggetggatattctcacaatagacccctaacatgcatc W G L N I F N V A G Y S H N R P L T C I

atgtatgctatattccaggaaagagacctcctaaagacattcagaatctcatctgacaca M Y A I F Q E R D L L K T F R I S S D T
tttataacctacatgatgactttagaagaccattaccattctgacgtggcatatcacaac F I T Y M M T L E D H Y H S D V A Y H N
agcctgcacgctgctgatgtagcccagtcgacccatgttctcctttctacaccagcatta S L H A A D V A Q S T H V L L S T P A L
gacgctgtcttcacagatttggaaatcctggctgccatttttgcagctgccatccatgac D A V F T D L E I L A A I F A A A I H D
gttgatcatcctggagtctccaatcagtttctcatcaacacaaattcagaacttgctttg V D H P G V S N Q F L I N T N S E L A L
atgtataatgatgaatctgtgttggaaaatcatcaccttgctgtgggtttcaaactgctg M Y N D E S V L E N H H L A V G F K L L
caagaagaacactgtgacatcttcatgaatctcaccaagaagcagcgtcagacactcagg Q E E H C D I F M N L T K K Q R Q T L R
aagatggttattgacatggtgttagcaactgatatgtctaaacacatgagcctgctggca K M V I D M V L A T D M S K H M S L L A
gacctgaagacaatggtagaaacgaagaaagttacaagttcaggcgttcttctcctagac D L K T M V E T K K V T S S G V L L L D
aactataccgatcgcattcaggtccttcgcaacatggtacactgtgcagacctgagcaac N Y T D R I Q V L R N M V H C A D L S N
cccaccaagtccttggaattgtatcggcaatggacagaccgcatcatggaggaatttttc P T K S L E L Y R Q W T D R I M E E F F
cagcagggagacaaagagcgggagaggggaatggaaattagcccaatgtgtgataaacac Q Q G D K E R E R G M E I S P M C D K H
acagcttctgtggaaaaatcccaggttggtttcatcgactacattgtccatccattgtgg T A S V E K S Q V G F I D Y I V H P L W
gagacatgggcagatttggtacagcctgatgctcaggacattctcgataccttagaagat E T W A D L V Q P D A Q D I L D T L E D
aacaggaactggtatcagagcatgatacctcaaagtccctcaccaccactggacgagcag N R N W Y Q S M I P Q S P S P P L D E Q
aacagggactgccagggtctgatggagaagtttcagtttgaactgactctcgatgaggaa N R D C Q G L M E K F Q F E L T L D E E
gattctgaaggacctgagaaggagggagagggacacagctatttcagcagcacaaagacg D S E G P E K E G E G H S Y F S S T K T
ctttgtgtgattgatccagaaaacagagattccctgggagagactgacatagacattgca L C V I D P E N R D S L G E T D I D I A
acagaagacaagtcccccgtggatacataatccccctctcgaggcatgcggtaccaagct T E D K S P V D T -t PCR primers:

[0630) PDE4D5:

P4478. pFastBac PDE4D5 M1-T745-X
tattccggattattcataccgtcccaccatcgggcgcggatctcggtccgaaacc atgtcgtactaccatcaccatcaccatcacgattacgatatcccaacgaccgaaaacctg M S Y Y H H H H H H D Y D I P T T E N L
tattttcagggccatatggctcagcagacaagcccggacactttaacagtacctgaagtg Y F Q G H M A Q Q T S P D T L T V P E V
gataatccgcattgtccaaacccgtggctgaacgaagaccttgtgaaatccttgcgagaa D N P H C P N P W L N E D L V K S L R E
aacctgttgcagcatgagaagtccaagacagcgaggaaatcggtttctcccaagctctct N L L Q H E K S K T A R K S V S P K L S
ccagtgatctctccgagaaattcccccaggcttctgcgcagaatgcttctcagcagcaac P V I S P R N S P R L L R R M L L S S N
atccccaaacagcggcgtttcacggtggcacatacatgttttgatgtggacaatggcaca I P K Q R R F T V A H T C F D V D N G T
tctgcgggacggagtcccttggatcccatgaccagcccaggatccgggctaattctccaa S A G R S P L D P M T S P G S G L I L Q
gcaaattttgtccacagtcaacgacgggagtccttcctgtatcgatccgacagcgattat A N F V H S Q R R E S F L Y R S D S D Y
gacctctctccaaagtctatgtcccggaactcctccattgccagtgatatacacggagat D L S P K S M S R N S S I A S D I H G D
gacttgattgtgactccatttgctcaggtcttggccagtctgcgaactgtacgaaacaac D L I V T P F A Q V L A S L R T V R N N
tttgctgcattaactaatttgcaagatcgagcacetagcaaaagatcacccatgtgcaac F A A L T N L Q D R A P S K R S P M C N
caaccatccatcaacaaagccaccataacagaggaggcctaccagaaactggccagcgag Q P S I N K A T I T E E A Y Q= K L A S E
accctggaggagctggactggtgtctggaccagctagagaccctacagaccaggcactcc T L E E L D W C L D Q L E T L Q T R H S
gtcagtgagatggcctccaacaagtttaaaaggatgcttaatcgggagctcacccatctc V S E M A S N K F K R M L N R E L T H L
tctgaaatgagtcggtctggaaatcaagtgtcagagtttatatcaaacacattcttagat S E M S R S G N Q V S E F I S N T F L D
aagcaacatgaagtggaaattccttctccaactcagaaggaaaaggagaaaaagaaaaga K Q H E V E I P S P T Q K E K E K K K R
ccaatgtctcagatcagtggagtcaagaaattgatgcacagctctagtctgactaattca P M S Q I S G V K K L M H S S S L T N S
agtatcccaaggtttggagttaaaactgaacaagaagatgtccttgccaaggaactagaa S I P R F G V K T E Q E D V L A K E L E
gatgtgaacaaatggggtcttcatgttttcagaatagcagagttgtctggtaaccggccc D V N K W G L H V F R I A E L S G N R P
ttgactgttatcatgcacaccatttttcaggaacgggatttattaaaaacatttaaaatt L T V I M H T I F Q E R D L L K T F K I
ccagtagatactttaattacatatcttatgactctcgaagaccattaccatgctgatgtg P V D T L I T Y L M T L E D H Y H A D V
gcctatcacaacaatatccatgctgcagatgttgtccagtctactcatgtgctattatct A Y H N N I H A A D V V Q S T H V L L S
acacctgctttggaggctgtgtttacagatttggagattcttgcagcaatttttgccagt T P A L E A V F T D L E I L A A I F A S
gcaatacatgatgtagatcatcctggtgtgtccaatcaatttctgatcaatacaaactct A I H D V D H P G V S N Q F L I N T N S
gaacttgccttgatgtacaatgattcctcagtcttagagaaccatcatttggctgtgggc E L A L M Y N D S S V L E N H H L A V G
tttaaattgcttcaggaagaaaactgtgacattttccagaatttgaccaaaaaacaaaga F K L L Q E E N C D I F Q N L T K K Q R
caatctttaaggaaaatggtcattgacatcgtacttgcaacagatatgtcaaaacacatg Q S L R K M V I D I V L A T D M S K H M
aatctactggctgatttgaagactatggttgaaactaagaaagtgacaagctctggagtt N L L A D L K T M V E T K K V T S S G V
cttcttcttgataattattccgataggattcaggttcttcagaatatggtgcactgtgca L L L D N Y S D R I Q V L Q N M V H C A
gatctgagcaacccaacaaagcctctccagctgtaccgccagtggacggaccggataatg D L S N P T K P L Q L Y R Q W T D R I M
gaggagttcttccgccaaggagaccgagagagggaacgtggcatggagataagccccatg E E F F R Q G D R E R E R G M E I S P M
tgtgacaagcacaatgcttccgtggaaaaatcacaggtgggcttcatagactatattgtt C D K H N A S V E K S Q V G F I D Y I V
catcccctctgggagacatgggcagacctcgtccaccctgacgcccaggatattttggac H P L W E T W A D L V H P D A Q D I L D
actttggaggacaatcgtgaatggtaccagagcacaatccctcagagcccctctcctgca T L E D N R E W Y Q S T I P Q S P S P A
cctgatgacccagaggagggccggcagggtcaaactgagaaattccagtttgaactaact P D D P E E G R Q G Q T E K F Q F E L T
ttagaggaagatggtgagtcagacacggaaaaggacagtggcagtcaagtggaagaagac L E E D G E S D T E K D S G S Q V E E D
actagctgcagtgactccaagactctttgtactcaagactcagagtctactgaaattccc T S C S D S K T L C T Q D S E S T E I P
cttgatgaacaggttgaagaggaggcagtaggggaagaagaggaaagccagcctgaagcc L D E Q V E E E A V G E E E E S Q P E A
tgtgtcatagatgatcgttctcctgacacgtaacagtcgactagagcctgcagtctegag C V I D D R S P D T -gcatgcggtaccaagctt PCR Primers:

Example 145. PDE4 IC50 Determinations [0631] IC50s were determined by Scintillation Proximity Assay (SPA). The principle of the assay is based on the fact that cAMP, the PDE4 substrate, binds weakly to Yittrium Silicate SPA beads, whereas AMP, the product of PDE4 hydrolysis binds strongly. Thus, the extent of PDE4 hydrolysis of a sample of [3H]cAMP can be measured because only the [3H]AMP
produced by PDE4 hydrolysis will bind to the SPA beads and produce a scintillation signal.
PDE4 enzymes used for IC50 assays:

[0632] PDE4B: The catalytic domain of human PDE4B from S 152-S528 with an N-terminal His6 tag and thrombin cleavage site, expressed in E. coli and purified by metal ion affinity chromatography. Enzyme was stored in 50% glycerol at -20 . See Example 144.

[0633] PDE4D: The catalytic domain of human PDE4B from S316-V692 with an N-terminal His6 tag and thrombin cleavage site, expressed in E. coli and purified by metal ion affinity chromatography. Enzyme was stored in 50% glycerol at -20 . See Example 144.
[0634] PDE4B2: see Example 144.

[0635] PDE4D5: see Example 144.
IC50 procedure:

[0636] Compounds tested (see Tables 5 for compounds and results) were 3-fold serially diluted 11 times in DMSO from a starting concentration of 4 mM or 40 M, depending on compound potency. 1 l of each dilution was transferred into duplicate wells of a white polystyrene 384-well assay plate (Coming #3710). In addition to the compound dilutions, each assay plate contained control wells with 1 l of DMSO (to define 0 %
enzyme inhibition) or 1 l of 200 M roflumilast (to define 100 % enzyme inhibition). Using a Beckman FX
robot, 10 l of [3H] cAMP (Amersham TRK559) at 2 mCi/ml in assay buffer (50 mM
Tris, pH
7.5; 8.3 mM MgC1; 1.7 mM EGTA; 0.01 % BSA) was transferred to each assay well.
Next, 10 l of PDE4 enzyme in assay buffer was added and the plates were shaken for 30 s. at 1000 rpm to start the cAMP hydrolysis reaction. The concentrations of enzyme used were:
PDE4B, 80 ng/ml; PDE4D, 4 ng/ml; PDE4B2, 2.5 l of 50% glycerol stock/ml; PDE4D5 0.083 1 of 50 %
glycerol stock/ml. Assay plates were covered and incubated for 30 min. at 30 C. Reactions were stopped by robotic addition of 10 1 of 5 mg/ml SPA beads (Amersham RPNQ0013) in 18 mM ZnSO4. The assay plates were covered with clear plastic film, centrifuged for 1 min. at 1000 RPM to settle the SPA beads, and counted using a Wallac TriLux scintillation counter.
IC50's were calculated from the raw assay data by non-linear regression curve fitting using the Assay Explorer software package from MDL.

Example 146. IC50 Determinations assessed by TNF alpha production upon stimulation of whole blood cultures with LPS.

[0637] Compounds were assayed to generate IC50 numbers based on the inhibition of TNF-a release from whole blood cultures, using the following assay protocol (see Table 5 for compounds tested and results). Inhibition of PDE4B results in the inhibition of TNF-a release by whole blood cultures stimulated with lipopolysaccharide (LPS). The measurement of TNF-a release was used to assess compounds as PDE4B inhibitors.

[0638] Compounds were provided in DMSO at 20 mM and 2 l per well was added to one row of a dilution plate. Added 98 1 of RPMI 1640 media with 2.5% heat inactivated FBS to each well containing compound. The same media with 2% DMSO was prepared and 60 1 was added to each of the empty wells in the dilution plate. The compound was serially diluted 1:3 (30 1 to 60 l of media) for a total of 8 concentrations per compound. Wells were also prepared with 50 M roflumilast and piclamilast as 100% inhibition controls, and 2% DMSO
in media was used as 0% inhibition control. A 20 1 aliquot of each sample was transferred to an assay plate in duplicate.

[0639] Human buffy coat was obtained from the Stanford Medical School Blood Center and diluted with 7 volumes of RPMI 1640 media with 1% penicillin/streptomycin and 2.5% heat inactivated FBS. A 160 l aliquot of the diluted blood was added to each well of the assay plates, mixed and incubated for 1 hour at 37 C in 5% COZ. LPS (Sigma catalog number L2637) that had been diluted to 1 mg/ml in PBS and stored as 20 l aliquots at -20 C was thawed and diluted 1000x to 1 g/ml. A 20 l aliquot of this was added to each sample (final concentration 100 ng/ml LPS) after the 1 hour incubation. A background sample was prepared without addition of LPS as well. The samples were mixed on a shaker for one minute at 900 RPM and incubated for 4 hours at 37 C in 5% CO2. The plates were then put on a shaker for one minute at 900 RPM, followed by centrifuging at 100 x g for 10 minutes, deceleration setting of 5. The top 75 l of supematant was carefully pipetted to a new plate and frozen at -20 C.

[0640] A 50 l aliquot of incubation buffer (Biosource International Immunoassay Kit:Human TNF-a catalog number KHC3011) was added to each well of a plate coated with monoclonal antibody specific for hTNF-a (Biosource kit). The supematant blood samples were thawed and a 50 1 aliquot along with 50 l of diluent buffer (Biosource kit) were added to incubation buffer and the samples were incubated for 2 hours at room temperature. The samples were washed 4 times with 300 Uwell wash buffer (Biosource kit). A 100 l aliquot of biotinylated anti-TNFalpha (Biosource kit) was added and the samples incubated for 1 hour at room temperature. The samples were washed 4 times with 300 l/well wash buffer. A 100 1 aliquot of Streptavidin-HRP solution (BioSource kit) was added and the samples incubated for 30 minutes at room temperature. The samples were washed 4 times with 300 1/well wash buffer. A 100 1 aliquot of Chromagen (Biosource kit) was added and the samples incubated in the dark for 30 minutes. A 100 1 aliquot of stop solution (2N H2SO4) was added to each sample and samples were read at 450 nm on a WallacVictor for 0.1 sec/well.

Example 147. Rat inhibition studies [0641] All studies were done with male rats CD (SD) IGS BR (Crl) (Charles River, France), which were grouped in to 5 animal groups. Compound doses were as indicated in Table 5.
[0642] At the end of the acclimatization period, the non-fasted rats were weighed, individually identified on the tail with a permanent marker and administered by oral (po) or interperitoneal (ip) route with either vehicle, reference or test compound in a volume of 10 mL/kg adapted to the body weight. The animals were gathered in groups of 5 animals in a polystyrene labeled cage with sawdust covered floors. 2-hours after vehicle, reference or test substance administration, rats received an intravenous (iv) injection of 0.1 mg/kg LPS in a volume of 1 mL/kg of body weight. 2 h after LPS challenge (or as indicated in Tables 3B and 4B), blood samples were collected into tubes without anticoagulant by retro-orbital puncture under gas (isoflurane) anesthesia. Samples were allowed to clot at room teinperature for 5 to min then put on ice until there were prepared by centrifugation (6000xg for 3 min at 4 C) and stored at -20 C until use. TNFa levels were measured in serum samples in duplicate by ELISA technique according to the manufacturer's procedure (Rat TNFa kit Quantikine M
(RTAOO, R&D System, France)). Data are reported as percent decrease in observed TNFa levels versus TNFa levels observed for vehicle dosed animal groups.

Example 148: Site-directed Mutagenesis of PDE4B

[0643] Mutagenesis of PDE4B can be carried out according to the following procedure as described in Molecular Biology: Current Innovations and Future Trends. Eds.
A.M. Griffin and H.G.Griffin. (1995) ISBN 1-898486-01-8, Horizon Scientific Press, PO Box 1, Wymondham, Norfolk, U.K., among others.

[0644] In vitro site-directed mutagenesis is an invaluable technique for studying protein structure-function relationships, gene expression and vector modification.
Several methods have appeared in the literature, but many of these methods require single-stranded DNA as the template. The reason for this, historically, has been the need for separating the complementary strands to prevent reannealing. Use of PCR in site-directed mutagenesis accomplishes strand separation by using a denaturing step to separate the complementing strands and allowing efficient polymerization of the PCR primers. PCR site-directed methods thus allow site-specific mutations to be incorporated in virtually any double-stranded plasmid; eliminating the need for M13-based vectors or single-stranded rescue.

[0645] It is often desirable to reduce the number of cycles during PCR when performing PCR-based site-directed mutagenesis to prevent clonal expansion of any (undesired) second-site mutations. Limited cycling which would result in reduced product yield, is offset by increasing the starting template concentration. A selection is used to reduce the number of parental molecules coming through the reaction. Also, in order to use a single PCR primer set, it is desirable to optimize the long PCR method. Further, because of the extendase activity of some thermostable polymerases it is often necessary to incorporate an end-polishing step into the procedure prior to end-to-end ligation of the PCR-generated product containing the incorporated mutations in one or both PCR primers.

[0646] The following protocol provides a facile method for site-directed mutagenesis and accomplishes the above desired features by the incorporation of the following steps: (i) increasing template concentration approximately 1000-fold over conventional PCR conditions;
(ii) reducing the number of cycles from 25-30 to 5-10; (iii) adding the restriction endonuclease DpnI (recognition target sequence: 5-Gm6ATC-3, where the A residue is metllylated) to select against parental DNA (note: DNA isolated from almost all cominon strains of E.
coli is Dam-methylated at the sequence 5-GATC-3); (iv) using Taq Extender in the PCR mix for increased reliability for PCR to 10 kb; (v) using Pfu DNA polymerase to polish the ends of the PCR
product, and (vi) efficient intramolecular ligation in the presence of T4 DNA
ligase.

[0647] Plasmid template DNA (approximately 0.5 pmole) is added to a PCR
cocktail containing, in 25 ul of lx mutagenesis buffer: (20 mM Tris HCI, pH 7.5; 8 mM
MgCl2; 40 ug/ml BSA); 12-20 pmole of each primer (one of which must contain a 5-prime phosphate), 250 uM each dNTP, 2.5 U Taq DNA polyinerase, 2.5 U of Taq Extender (Stratagene).

[0648] The PCR cycling parameters are 1 cycle of: 4 min at 94 C, 2 min at 50 C
and 2 min at 72 C; followed by 5-10 cycles of 1 min at 94 C, 2 min at 54 C and 1 min at 72 C (step 1).
[0649] The parental template DNA and the linear, mutagenesis-primer incorporating newly synthesized DNA are treated with Dpnl (10 U) and Pfu DNA polymerase (2.5U).
This results in the DpnI digestion of the in vivo methylated parental template and hybrid DNA and the removal, by Pfu DNA polymerase, of the Taq DNA polymerase-extended base(s) on the linear PCR product.

[0650] The reaction is incubated at 37 C for 30 min and then transferred to 72 C for an additional 30 min (step 2).

[0651] Mutagenesis buffer (lx, 115 ul, containing 0.5 mM ATP) is added to the DpnI-digested, Pfu DNA polymerase-polished PCR products.

[0652] The solution is mixed and 10 ul is removed to a new microfuge tube and ligase (2-4 U) added.

[0653] The ligation is incubated for greater than 60 min at 37 C (step 3).
[0654] The treated solution is transformed into competent E. coli (step 4).

[0655] In addition to the PCR-based site-directed mutagenesis described above, other methods are available. Examples include those described in Kunkel (1985) Proc.
Natl. Acad.
Sci. 82:488-492; Eckstein et al. (1985) Nucl. Acids Res. 13:8764-8785; and using the GeneEditorTM Site-Directed Mutageneis Sytem from Promega.

[0656] All patents and other references cited in the specification are indicative of the level of skill of those skilled in the art to which the invention pertains, and are incorporated by reference in their entireties, including any tables and figures, to the same extent as if each reference had been incorporated by reference in its entirety individually.

[0657] One skilled in the art would readily appreciate that the present invention is well adapted to obtain the ends and advantages mentioned, as well as those inlierent therein. The methods, variances, and compositions described herein as presently representative of preferred embodiments are exeinplary and are not intended as limitations on the scope of the invention.
Changes therein and other uses will occur to those skilled in the art, which are encompassed within the spirit of the invention, are defined by the scope of the claims.

[0658] It will be readily apparent to one skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention. For example, variations can be made to crystallization or co-crystallization conditions for PDE4B proteins and/or various phosphodiesterase domain sequences can be used. Thus, such additional embodiments are within the scope of the present invention and the following claims.

[0659] The invention illustratively described herein suitably may be practiced in the absence of any element or elements, limitation or limitations which is not specifically disclosed herein.
Thus, for example, in each instance herein any of the terms "comprising", "consisting essentially of' and "consisting of' may be replaced with either of the other two terms. The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments and optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the appended claims.

[0660] In addition, where features or aspects of the invention are described in terms of Markush groups or other grouping of alternatives, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group or other group.

[0661] Also, unless indicated to the contrary, where various numerical values are provided for embodiments, additional embodiments are described by taking any 2 different values as the endpoints of a range. Such ranges are also within the scope of the described invention.

[0662] Thus, additional embodiments are within the scope of the invention and within the following claims.

Table 1 REMARK Written by 0 version 8Ø11 CRYSTI 1.000 1.000 1.000 90.00 90.00 90.00 ORIGXI 1.000000 0.000000 0.000000 0.00000 ORIGX2 0.000000 1.000000 0.000000 0.00000 ORIGX3 0.000000 0.000000 1.000000 0.00000 SCALE1 1.000000 -0.000026 -0.000026 0.00000 SCALE2 0.000000 1.000000 -0.000026 0.00000 SCALE3 0.000000 0.000000 1.000000 0.00000 ATOM 1 CB ARG A 155 79.971 64.880 65.907 1.00 23.05 A
ATOM 2 CG ARG A 155 80.613 63.882 66.857 1.00 22.25 A
ATOM 3 CD ARG A 155 81.907 63.327 66.300 1.00 21.47 A
ATOM 4 NE ARG A 155 82.378 62.172 67.060 1.00 22.64 A
ATOM 5 CZ ARG A 155 82.051 60.910 66.792 1.00 20.30 A
ATOM 6 NH1 ARG A 155 81.248 60.626 65.775 1.00 20.18 A
ATOM 7 NH2 ARG A 155 82.533 59.927 67.542 1.00 21.88 A
ATOM 8 C ARG A 155 80.084 66.914 64.494 1.00 22.90 A
ATOM 9 0 ARG A 155 80.453 66.813 63.325 1.00 22.93 A
ATOM 10 N ARG A 155 80.953 66.971 66.838 1.00 22.95 A
ATOM 11 CA ARG A 155 80.788 66.143 65.606 1.00 22.82 A
ATOM 12 N PHE A 156 79.060 67.679 64.859 1.00 22.91 A
ATOM 13 CA PHE A 156 78.296 68.439 63.879 1.00 22.30 A
ATOM 14 CB PHE A 156 76.906 68,746 64.434 1.00 22.43 A
ATOM 15 CG PHE A 156 75.997 67.556 64.464 1.00 22.09 A
ATOM 16 CD1 PHE A 156 75.372 67.121 63.306 1.00 22.60 A
ATOM 17 CD2 PHE A 156 75.786 66.858 65.641 1.00 23.79 A
ATOM 18 CE1 PHE A 156 74.550 66.009 63.325 1.00 23.14 A
ATOM 19 CE2 PHE A 156 74.966 65.747 65.667 1.00 23.39 A
ATOM 20 CZ PHE A 156 74.348 65.322 64.506 1.00 23.54 A
ATOM 21 C PHE A 156 78.966 69.728 63.423 1.00 23.31 A
ATOM 22 0 PHE A 156 78.749 70.177 62.300 1.00 23.75 A
ATOM 23 N GLY A 157 79.776 70.321 64.295 1.00 22.97 A
ATOM 24 CA GLY A 157 80.459 71.552 63.943 1.00 24.05 A
ATOM 25 C GLY A 157 79.526 72.691 63.579 1.00 24.76 A
ATOM 26 0 GLY A 157 79.888 73.562 62.786 1.00 26.86 A
ATOM 27 N VAL A 158 78.328 72.693 64.157 1.00 25.15 A
ATOM 28 CA VAL A 158 77.349 73.742 63.880 1.00 24.88 A
ATOM 29 CB VAL A 158 76.019 73.475 64.612 1.00 24.20 A
ATOM 30 CG1 VAL A 158 75.028 74.583 64.304 1.00 23.49 A
ATOM 31 CG2 VAL A 158 75.453 72.128 64.191 1.00 25.00 A
ATOM 32 C VAL A 158 77.876 75.105 64.319 1.00 26.65 A
ATOM 33 0 VAL A 158 78.278 75,286 65.471 1.00 27.38 A
ATOM 34 N ASN A 159 77.872 76.059 63.392 1.00 27.36 A
ATOM 35 CA ASN A 159 78.351 77.407 63.669 1.00 28.38 A
ATOM 36 C ASN A 159 77.553 78.037 64.806 1.00 29.46 A
ATOM 37 0 ASN A 159 76.329 77.907 64.867 1.00 29.12 A
ATOM 38 CB ASN A 159 78.275 78.271 62.410 1.00 28.09 A
ATOM 39 CG ASN A 159 79.112 77.719 61.273 1.00 20.00 A
ATOM 40 OD1 ASN A 159 80.222 77.234 61.486 1.00 20.00 A
ATOM 41 ND2 ASN A 159 78.581 77.793 60.059 1.00 20.00 A
ATOM 42 N THR A 160 78.258 78.720 65.701 1.00 27.70 A
ATOM 43 CA THR A 160 77.631 79.364 66.849 1.00 27.94 A
ATOM 44 C THR A 160 76.431 80.212 66.451 1.00 28.14 A
ATOM 45 0 THR A 160 75.420 80.236 67.152 1.00 28.28 A
ATOM 46 CB THR A 160 78.615 80.269 67.590 1.00 26.10 A
ATOM 47 001 THR A 160 79.724 79.492 68.058 1.00 20.00 A
ATOM 48 CG2 THR A 160 77.938 80.929 68.782 1.00 20.00 A
ATOM 49 N GLU A 161 76.543 80.906 65.323 1.00 27.39 A
ATOM 50 CA GLU A 161 75.466 81.765 64.847 1.00 28.03 A
ATOM 51 C GLU A 161 74.271 80.974 64.322 1.00 27.84 A
ATOM 52 0 GLU A 161 73.218 81.546 64.041 1.00 28.36 A
ATOM 53 CB GLU A 161 75.970 82.692 63.740 1.00 28.29 A
ATOM 54 CG GLU A 161 77.100 83.612 64.168 1.00 20.00 A
ATOM 55 CD GLU A 161 77.573 84.515 63.046 1.00 20.00 A

ATOM 56 OE1 GLU A 161 77.016 84.420 61.933 1.00 20.00 A
ATOM 57 OE2 GLU A 161 78.500 85.317 63.280 1.00 20.00 A
ATOM 58 N ASN A 162 74.431 79.662 64.191 1.00 26.56 A
ATOM 59 CA ASN A 162 73.348 78.824 63.688 1.00 26.62 A
ATOM 60 CB ASN A 162 73.838 77.996 62.497 1.00 25.54 A
ATOM 61 CG ASN A 162 74.307 78.863 61.343 1.00 25.39 A
ATOM 62 OD1 ASN A 162 73.613 79.796 60.935 1.00 27.84 A
ATOM 63 ND2 ASN A 162 75.486 78.561 60.811 1.00 27.27 A
ATOM 64 C ASN A 162 72.762 77.911 64.757 1.00 26.85 A
ATOM 65 0 ASN A 162 71.782 77.202 64.512 1.00 27.85 A
ATOM 66 N GLU A 163 73.356 77.938 65.945 1.00 26.44 A
ATOM 67 CA GLU A 163 72.887 77.118 67.054 1.00 26.49 A
ATOM 68 CB GLU A 163 73.726 77.382 68.306 1.00 26.50 A
ATOM 69 CG GLU A 163 74.517 76.184 68.788 1.00 28.58 A
ATOM 70 CD GLU A 163 75.856 76.049 68.098 1,00 29.98 A
ATOM 71 OE1 GLU A 163 75.896 76.082 66.849 1.00 32.90 A
ATOM 72 OE2 GLU A 163 76.874 75.904 68.810 1.00 31.77 A
ATOM 73 C GLU A 163 71.425 77.396 67.375 1.00 25.70 A
ATOM 74 0 GLU A 163 70.598 76.481 67.392 1.00 25.71 A
ATOM 75 N ASP A 164 71.116 78.665 67.636 1.00 25.48 A
ATOM 76 CA ASP A 164 69.759 79.084 67.968 1.00 25.27 A
ATOM 77 CB ASP A 164 69.740 80.566 68.359 1.00 27.36 A
ATOM 78 CG ASP A 164 70.568 80.854 69.597 1.00 29.80 A
ATOM 79 OD1 ASP A 164 70.276 80.264 70.659 1.00 32.25 A
ATOM 80 OD2 ASP A 164 71.508 81.674 69.508 1.00 32.26 A
ATOM 81 C ASP A 164 68.777 78.862 66,827 1.00 24.21 A
ATOM 82 0 ASP A 164 67.622 78.506 67.056 1.00 24.88 A
ATOM 83 N HIS A 165 69.234 79.081 65.599 1.00 22.75 A
ATOM 84 CA HIS A 165 68.371 78.906 64.438 1.00 23.18 A
ATOM 85 CB HIS A 165 69.054 79.453 63.184 1.00 24.09 A
ATOM 86 CG HIS A 165 69.299 80.930 63.228 1.00 25.46 A
ATOM 87 CD2 HIS A 165 70.440 81.648 63.102 1.00 26.59 A
ATOM 88 ND1 HIS A 165 68.291 81.846 63.443 1.00 27.36 A
ATOM 89 CE1 HIS A 165 68.802 83.065 63.449 1.00 26.84 A
ATOM 90 NE2 HIS A 165 70.104 82.972 63.245 1.00 27.21 A
ATOM 91 C HIS A 165 68.014 77.438 64.244 1.00 22.85 A
ATOM 92 0 HIS A 165 66.886 77.110 63.880 1.00 23.63 A
ATOM 93 N LEU A 166 68.977 76.558 64.499 1.00 22.36 A
ATOM 94 CA LEU A 166 68.755 75.122 64.360 1.00 22.41 A
ATOM 95 CB LEU A 166 70.078 74.362 64.487 1.00 22.69 A
ATOM 96 CG LEU A 166 70.342 73.168 63.563 1.00 24.72 A
ATOM 97 CD1 LEU A 166 71.318 72.236 64.258 1.00 24.38 A
ATOM 98 CD2 LEU A 166 69.052 72.430 63.226 1.00 23.56 A
ATOM 99 C LEU A 166 67.793 74.633 65.446 1.00 22.38 A
ATOM 100 0 LEU A 166 66.851 73.896 65.165 1.00 21.57 A
ATOM 101 N ALA A 167 68.032 75.042 66.689 1.00 22.78 A
ATOM 102 CA ALA A 167 67.175 74.624 67.796 1.00 22.78 A
ATOM 103 CB ALA A 167 67.727 75.156 69.117 1.00 23.00 A
ATOM 104 C ALA A 167 65.739 75.104 67.595 1.00 22.72 A
ATOM 105 0 ALA A 167 64.784 74.422 67.968 1.00 23.75 A
ATOM 106 N LYS A 168 65.595 76.287 67.006 1.00 23.26 A
ATOM 107 CA LYS A 168 64.281 76.857 66.738 1.00 24.03 A
ATOM 108 CB LYS A 168 64.444 78.258 66.145 1.00 23.60 A
ATOM 109 CG LYS A 168 63.155 78.955 65.751 1.00 25.93 A
ATOM 110 CD LYS A 168 63.468 80.312 65.117 1.00 27.10 A
ATOM 111 CE LYS A 168 64.440 80.154 63.949 1.00 29.33 A
ATOM 112 NZ LYS A 168 64.847 81.450 63.326 1.00 29.14 A
ATOM 113 C LYS A 168 63.524 75.958 65.760 1.00 22.83 A
ATOM 114 0 LYS A 168 62.334 75.700 65.932 1.00 24.33 A
ATOM 115 N GLU A 169 64.225 75.483 64.735 1.00 23.15 A
ATOM 116 CA GLU A 169 63.614 74.611 63.735 1.00 22.21 A
ATOM 117 CB GLU A 169 64.583 74.350 62.572 1.00 21.88 A
ATOM 118 CG GLU A 169 65.003 75.569 61.750 1.00 25.15 A
ATOM 119 CD GLU A 169 63.844 76.254 61.043 1.00 25.44 A
ATOM 120 OE1 GLU A 169 62.900 75.557 60.607 1.00 28.94 A
ATOM 121 OE2 GLU A 169 63.889 77.497 60.904 1.00 28.04 A
ATOM 122 C GLU A 169 63.229 73.272 64.356 1.00 22.48 A

ATOM 123 0 GLU A 169 62.154 72.737 64.088 1.00 23.05 A
ATOM 124 N LEU A 170 64.114 72.736 65.192 1.00 21.62 A
ATOM 125 CA LEU A 170 63.878 71.447 65.824 1.00 21.58 A
ATOM 126 CB LEU A 170 65.175 70.934 66.457 1.00 20.64 A
ATOM 127 CG LEU A 170 66.293 70.607 65.460 1.00 21.61 A
ATOM 128 CD1 LEU A 170 67.567 70.244 66.211 1.00 21.59 A
ATOM 129 CD2 LEU A 170 65.853 69.461 64.554 1.00 21.25 A
ATOM 130 C LEU A 170 62.742 71.415 66.843 1.00 21.28 A
ATOM 131 0 LEU A 170 62.343 70.342 67.293 1.00 22.17 A
ATOM 132 N GLU A 171 62.216 72.578 67.214 1.00 21.88 A
ATOM 133 CA GLU A 171 61.105 72.602 68.160 1.00 22.83 A
ATOM 134 CB GLU A 171 60.757 74.035 68.572 1.00 23.48 A
ATOM 135 CG GLU A 171 61.792 74.710 69.453 1.00 25.17 A
ATOM 136 CD GLU A 171 61.249 75.956 70.128 1.00 28.89 A
ATOM 137 OE1 GLU A 171 60.748 76.853 69.415 1.00 31.29 A
ATOM 138 OE2 GLU A 171 61.322 76.037 71.373 1.00 31.71 A
ATOM 139 C GLU A 171 59.887 71.959 67.501 1.00 22.17 A
ATOM 140 0 GLU A 171 58.942 71.553 68.175 1.00 22.98 A
ATOM 141 N ASP A 172 59.928 71.865 66.175 1.00 21_.31 A
ATOM 142 CA ASP A 172 58.838 71.278 65.404 1.00 20.32 A
ATOM 143 CB ASP A 172 58.596 72.116 64.149 1.00 21.90 A
ATOM 144 CG ASP A 172 58.305 73.568 64.476 1.00 26.96 A
ATOM 145 OD1 ASP A 172 57.361 73.815 65.257 1.00 27.29 A
ATOM 146 OD2 ASP A 172 59.021 74.456 63.962 1.00 29.99 A
ATOM 147 C ASP A 172 59.128 69.826 65.017 1.00 18.25 A
ATOM 148 0 ASP A 172 58.494 69.274 64.122 1.00 19.05 A
ATOM 149 N LEU A 173 60.078 69.214 65.714 1.00 17.76 A
ATOM 150 CA LEU A 173 60.468 67.831 65.458 1.00 17.09 A
ATOM 151 CB LEU A 173 61.434 67.353 66.546 1.00 17.23 A
ATOM 152 CG LEU A 173 61.953 65.918 66.406 1.00 18.22 A
ATOM 153 CD1 LEU A 173 62.842 65.820 65.174 1.00 18.47 A
ATOM 154 CD2 LEU A 173 62.726 65.525 67.663 1.00 19.38 A
ATOM 155 C LEU A 173 59.290 66.863 65.380 1.00 17.63 A
ATOM 156 0 LEU A 173 59.252 65.991 64.511 1.00 18.15 A
ATOM 157 N ASN A 174 58.328 67.013 66.285 1.00 17.68 A
ATOM 158 CA ASN A 174 57.177 66.118 66.310 1.00 18.34 A
ATOM 159 CB ASN A 174 56.713 65.899 67.754 1.00 20.61 A
ATOM 160 CG ASN A 174 57.847 65.512 68.682 1.00 22.17 A
ATOM 161 OD1 ASN A 174 58.672 64.667 68.347 1.00 22.67 A
ATOM 162 ND2 ASN A 174 57.886 66.124 69.865 1.00 23.82 A
ATOM 163 C ASN A 174 55.985 66.591 65.481 1.00 18.49 A
ATOM 164 0 ASN A 174 54.876 66.089 65.657 1.00 19.70 A
ATOM 165 N LYS A 175 56.212 67.534 64.571 1.00 18.41 A
ATOM 166 CA LYS A 175 55.131 68.078 63.748 1.00 18.38 A
ATOM 167 CB LYS A 175 55.036 69.591 63.945 1.00 20.31 A
ATOM 168 CG LYS A 175 54.789 70.030 65.379 1.00 22.09 A
ATOM 169 CD LYS A 175 54.812 71.545 65.469 1.00 23.35 A
ATOM 170 CE LYS A 175 54.558 72.034 66.883 1.00 26.61 A
ATOM 171 NZ LYS A 175 54.612 73.524 66.946 1.00 28.25 A
ATOM 172 C LYS A 175 55.310 67.790 62.263 1.00 17.31 A
ATOM 173 0 LYS A 175 56.430 67.706 61.768 1.00 17.36 A
ATOM 174 N TRP A 176 54.191 67.657 61.556 1.00 18.04 A
ATOM 175 CA TRP A 176 54.206 67.390 60.122 1.00 17.86 A
ATOM 176 CB TRP A 176 52.776 67.092 59.648 1.00 17.14 A
ATOM 177 CG TRP A 176 52.654 66.297 58.372 1.00 15.60 A
ATOM 178 CD2 TRP A 176 53.051 64.933 58.162 1.00 15.09 A
ATOM 179 CE2 TRP A 176 52.659 64.579 56.855 1.00 14.34 A
ATOM 180 CE3 TRP A 176 53.697 63.977 58.953 1.00 15.37 A
ATOM 181 CD1 TRP A 176 52.060 66.703 57.208 1.00 15.68 A
ATOM 182 NE1 TRP A 176 52.057 65.676 56.295 1.00 14.60 A
ATOM 183 CZ2 TRP A 176 52.889 63.311 56.323 1.00 14.99 A
ATOM 184 CZ3 TRP A 176 53.925 62.717 58.423 1.00 16.21 A
ATOM 185 CH2 TRP A 176 53.521 62.395 57.121 1.00 15.03 A
ATOM 186 C TRP A 176 54.757 68.622 59.402 1.00 18.06 A
ATOM 187 0 TRP A 176 55.260 68.525 58.283 1.00 18.52 A
ATOM 188 N GLY A 177 54.686 69.773 60.068 1.00 19.48 A
ATOM 189 CA GLY A 177 55.153 71.016 59.477 1.00 20.36 A

ATOM 190 C GLY A 177 56.632 71.350 59.577 1.00 20.93 A
ATOM 191 0 GLY A 177 57.061 72.396 59.094 1.00 22.82 A
ATOM 192 N LEU A 178 57.419 70.482 60.202 1.00 19.75 A
ATOM 193 CA LEU A 178 58.854 70.724 60.323 1.00 18.09 A
ATOM 194 CB LEU A 178 59.529 69.499 60.950 1.00 18.68 A
ATOM 195 CG LEU A 178 61.044 69.487 61.175 1.00 18.87 A
ATOM 196 CD1 LEU A 178 61.380 68.476 62.253 1.00 22.51 A
ATOM 197 CD2 LEU A 178 61.767 69.143 59.886 1.00 19.56 A
ATOM 198 C LEU A 178 59.450 71.012 58.945 1.00 17.28 A
ATOM 199 0 LEU A 178 59.044 70.407 57.953 1.00 19.08 A
ATOM 200 N ASN A 179 60.402 71.940 58.879 1.00 17.34 A
ATOM 201 CA ASN A 179 61.037 72.285 57.607 1.00 17.56 A
ATOM 202 CB ASN A 179 61.126 73.803 57.440 1.00 18.95 A
ATOM 203 CG ASN A 179 61.540 74.204 56.042 1.00 19.70 A
ATOM 204 OD1 ASN A 179 62.490 73.654 55.486 1.00 21.00 A
ATOM 205 ND2 ASN A 179 60.831 75.167 55.464 1.00 21.60 A
ATOM 206 C ASN A 179 62.436 71.696 57.566 1.00 17.32 A
ATOM 207 0 ASN A 179 63.385 72.287 58.085 1.00 17.55 A
ATOM 208 N ILE A 180 62.563 70.537 56.930 1.00 17.15 A
ATOM 209 CA ILE A 180 63.845 69.851 56.859 1.00 16.59 A
ATOM 210 CB ILE A 180 63.655 68.418 56.316 1.00 16.15 A
ATOM 211 CG2 ILE A 180 63.407 68.457 54.818 1.00 16.84 A
ATOM 212 CG1 ILE A 180 64.876 67.563 56.658 1.00 16.30 A
ATOM 213 CD1 ILE A 180 65.048 67.323 58.152 1.00 18.04 A
ATOM 214 C ILE A 180 64.883 70.605 56.027 1.00 15.54 A
ATOM 215 0 ILE A 180 66.085 70.429 56.223 1.00 16.30 A
ATOM 216 N PHE A 181 64.416 71.449 55.110 1.00 15.72 A
ATOM 217 CA PHE A 181 65.307 72.244 54.271 1.00 16.55 A
ATOM 218 CB PHE A 181 64.499 73.038 53.239 1.00 16.12 A
ATOM 219 CG PHE A 181 63.813 72.176 52.212 1.00 17.03 A
ATOM 220 CD1 PHE A 181 64.529 71.609 51.170 1.00 18.63 A
ATOM 221 CD2 PHE A 181 62.459 71.907 52.313 1.00 17.82 A
ATOM 222 CE1 PHE A 181 63.903 70.786 50.247 1.00 20.00 A
ATOM 223 CE2 PHE A 181 61.829 71.085 51.395 1.00 17.51 A
ATOM 224 CZ PHE A 181 62.554 70.523 50.360 1.00 17.36 A
ATOM 225 C PHE A 181 66.109 73.206 55.146 1.00 15.85 A
ATOM 226 0 PHE A 181 67.302 73.410 54.924 1.00 17.31 A
ATOM 227 N ASN A 182 65.456 73.794 56.143 1.00 17.65 A
ATOM 228 CA ASN A 182 66.161 74.713 57.027 1.00 18.69 A
ATOM 229 CB ASN A 182 65.183 75.544 57.854 1.00 19.86 A
ATOM 230 CG ASN A 182 64.335 76.457 56.997 1.00 21.86 A
ATOM 231 OD1 ASN A 182 64.759 76.892 55.924 1.00 23.94 A
ATOM 232 ND2 ASN A 182 63.134 76.765 57.473 1.00 23.53 A
ATOM 233 C ASN A 182 67.089 73.935 57.944 1.00 19.19 A
ATOM 234 0 ASN A 182 68.163 74.417 58.302 1.00 20.79 A
ATOM 235 N VAL A 183 66.678 72.731 58.328 1.00 18.83 A
ATOM 236 CA VAL A 183 67.522 71.901 59.184 1.00 17.82 A
ATOM 237 CB VAL A 183 66.815 70.580 59.555 1.00 17.94 A
ATOM 238 CG1 VAL A 183 67.799 69.624 60.219 1.00 18.47 A
ATOM 239 CG2 VAL A 183 65.639 70.868 60.473 1.00 17.83 A
ATOM 240 C VAL A 183 68.829 71.585 58.457 1.00 17.58 A
ATOM 241 0 VAL A 183 69.907 71.628 59.048 1.00 18.65 A
ATOM 242 N ALA A 184 68.734 71.274 57.167 1.00 17.48 A
ATOM 243 CA ALA A 184 69.918 70.959 56.377 1.00 17.45 A
ATOM 244 CB ALA A 184 69.505 70.479 54.985 1.00 16.66 A
ATOM 245 C ALA A 184 70.819 72.182 56.262 1.00 17.80 A
ATOM 246 0 ALA A 184 72.047 72.079 56.329 1.00 19.06 A
ATOM 247 N GLY A 185 70.201 73.347 56.104 1.00 18.60 A
ATOM 248 CA GLY A 185 70.965 74.571 55.971 1.00 19.69 A
ATOM 249 C GLY A 185 71.758 74.958 57.203 1.00 20.68 A
ATOM 250 0 GLY A 185 72.854 75.502 57.087 1.00 22.71 A
ATOM 251 N TYR A 186 71.217 74.678 58.383 1.00 20.97 A
ATOM 252 CA TYR A 186 71.897 75.031 59.627 1.00 21.00 A
ATOM 253 CB TYR A 186 70.870 75.458 60.685 1.00 20.43 A
ATOM 254 CG TYR A 186 70.115 76.730 60.348 1.00 21.83 A
ATOM 255 CD1 TYR A 186 70.793 77.917 60.100 1.00 21.59 A
ATOM 256 CE1 TYR A 186 70.110 79.085 59.798 1.00 24.25 A

ATOM 257 CD2 TYR A 186 68.730 76.745 60.286 1.00 22.13 A
ATOM 258 CE2 TYR A 186 68.035 77.910 59.984 1.00 23.32 A
ATOM 259 CZ TYR A 186 68.733 79.076 59.741 1.00 24.49 A
ATOM 260 OH TYR A 186 68.055 80.239 59.441 1.00 27.87 A
ATOM 261 C TYR A 186 72.776 73.918 60.195 1.00 21.63 A
ATOM 262 0 TYR A 186 73.418 74.103 61.229 1.00 23.64 A
ATOM 263 N SER A 187 72.819 72.769 59.526 1.00 20.69 A
ATOM 264 CA SER A 187 73.631 71.655 60.017 1.00 19.57 A
ATOM 265 CB SER A 187 72.745 70.432 60.265 1.00 18.38 A
ATOM 266 OG SER A 187 72.187 69.951 59.048 1.00 18.57 A
ATOM 267 C SER A 187 74.767 71.261 59.081 1.00 20.33 A
ATOM 268 0 SER A 187 75.240 70.126 59.120 1.00 20.31 A
ATOM 269 N HIS A 188 75.216 72.195 58.248 1.00 20.86 A
ATOM 270 CA HIS A 188 76.293 71.917 57.304 1.00 21.53 A
ATOM 271 CB HIS A 188 77.581 71.576 58.051 1.00 22.29 A
ATOM 272 CG HIS A 188 78.127 72.719 58.844 1.00 23.94 A
ATOM 273 CD2 HIS A 188 78.417 72.835 60.162 1.00 24.48 A
ATOM 274 ND1 HIS A 188 78.440 73.934 58.273 1.00 25.01 A
ATOM 275 CE1 HIS A 188 78.899 74.750 59.205 1.00 25.13 A
ATOM 276 NE2 HIS A 188 78.896 74.108 60.360 1.00 24.90 A
ATOM 277 C HIS A 188 75.913 70.776 56.373 1.00 19.79 A
ATOM 278 0 HIS A 188 76.726 69.906 56.058 1.00 20.06 A
ATOM 279 N ASN A 189 74.657 70.791 55.949 1.00 19.07 A
ATOM 280 CA ASN A 189 74.116 69.793 55.044 1.00 18.96 A
ATOM 281 CB ASN A 189 74.795 69.895 53.673 1.00 20.86 A
ATOM 282 CG ASN A 189 74.035 69.143 52.591 1.00 24.63 A
ATOM 283 OD1 ASN A 189 72.823 69.312 52.439 1.00 28.24 A
ATOM 284 ND2 ASN A 189 74.745 68.312 51.831 1.00 26.51 A
ATOM 285 C ASN A 189 74.207 68.365 55.569 1.00 17.10 A
ATOM 286 0 ASN A 189 74.555 67.442 54.834 1.00 18.62 A
ATOM 287 N ARG A 190 73.893 68.191 56.849 1.00 16.40 A
ATOM 288 CA ARG A 190 73.878 66.870 57.466 1.00 15.56 A
ATOM 289 CB ARG A 190 74.990 66.743 58.513 1.00 17.82 A
ATOM 290 CG ARG A 190 76.401 66.702 57.926 1.00 19.21 A
ATOM 291 CD ARG A 190 76.527 65.600 56.875 1.00 22.29 A
ATOM 292 NE ARG A 190 77.918 65.295 56.546 1.00 24.25 A
ATOM 293 CZ ARG A 190 78.295 64.497 55.551 1.00 23.85 A
ATOM 294 NH1 ARG A 190 77.383 63.920 54.773 1.00 23.57 A
ATOM 295 NH2 ARG A 190 79.586 64.270 55.336 1.00 25.08 A
ATOM 296 C ARG A 190 72.507 66.686 58.126 1.00 14.09 A
ATOM 297 0 ARG A 190 72.404 66.341 59.302 1.00 14.55 A
ATOM 298 N PRO A 191 71.426 66.894 57.356 1.00 12.43 A
ATOM 299 CD PRO A 191 71.393 67.152 55.906 1.00 12.36 A
ATOM 300 CA PRO A 191 70.068 66.756 57.887 1.00 12.25 A
ATOM 301 CB PRO A 191 69.193 67.206 56.719 1.00 10.84 A
ATOM 302 CG PRO A 191 69.984 66.732 55.536 1.00 11.01 A
ATOM 303 C PRO A 191 69.681 65.373 58.391 1.00 11.81 A
ATOM 304 0 PRO A 191 68.944 65.260 59.363 1.00 12.80 A
ATOM 305 N LEU A 192 70.160 64.320 57.739 1.00 12.56 A
ATOM 306 CA LEU A 192 69.799 62.981 58.186 1.00 12.47 A
ATOM 307 CB LEU A 192 70.233 61.936 57.154 1.00 12.79 A
ATOM 308 CG LEU A 192 69.905 60.480 57.487 1.00 13.72 A
ATOM 309 CD1 LEU A 192 68.399 60.294 57.658 1.00 12.53 A
ATOM 310 CD2 LEU A 192 70.437 59.592 56.376 1.00 15.24 A
ATOM 311 C LEU A 192 70.408 62.659 59.544 1.00 12.15 A
ATOM 312 0 LEU A 192 69.730 62.130 60.422 1.00 13.07 A
ATOM 313 N THR A 193 71.685 62.983 59.724 1.00 13.19 A
ATOM 314 CA THR A 193 72.336 62.711 60.998 1.00 13.74 A
ATOM 315 CB THR A 193 73.855 62.968 60.907 1.00 14.07 A
ATOM 316 OG1 THR A 193 74.427 62.083 59.936 1.00 15.87 A
ATOM 317 CG2 THR A 193 74.524 62.717 62.253 1.00 15.95 A
ATOM 318 C THR A 193 71.725 63.584 62.088 1.00 13.16 A
ATOM 319 0 THR A 193 71.447 63.114 63.191 1.00 13.24 A
ATOM 320 N CYS A 194 71.505 64.854 61.768 1.00 13.77 A
ATOM 321 CA CYS A 194 70.920 65.789 62.716 1.00 14.71 A
ATOM 322 CB CYS A 194 70.856 67.184 62.089 1.00 16.72 A
ATOM 323 SG CYS A 194 70.205 68.462 63.179 1.00 21.33 A

ATOM 324 C CYS A 194 69.519 65.355 63.150 1.00 14.33 A
ATOM 325 0 CYS A 194 69.229 65.250 64.344 1.00 14.68 A
ATOM 326 N ILE A 195 68.646 65.090 62.184 1.00 13.98 A
ATOM 327 CA ILE A 195 67.286 64.701 62.520 1.00 14.07 A
ATOM 328 CB ILE A 195 66.361 64.719 61.264 1.00 14.75 A
ATOM 329 CG2 ILE A 195 66.402 63.384 60.535 1.00 16.38 A
ATOM 330 CG1 ILE A 195 64.926 65.024 61.693 1.00 17.02 A
ATOM 331 CD1 ILE A 195 64.765 66.403 62.288 1.00 18.15 A
ATOM 332 C ILE A 195 67.230 63.335 63.202 1.00 12.83 A
ATOM 333 0 ILE A 195 66.420 63.126 64.107 1.00 14.09 A
ATOM 334 N MET A 196 68.098 62.407 62.801 1.00 13.45 A
ATOM 335 CA MET A 196 68.081 61.091 63.431 1.00 11.19 A
ATOM 336 CB MET A 196 68.973 60.106 62.669 1.00 12.34 A
ATOM 337 CG MET A 196 68.335 59.567 61.381 1.00 12.99 A
ATOM 338 SD MET A 196 66.881 58.516 61.676 1.00 14.52 A
ATOM 339 CE MET A 196 67.676 56.931 62.036 1.00 15.62 A
ATOM 340 C MET A 196 68.519 61.175 64.894 1.00 12.03 A
ATOM 341 0 MET A 196 67.968 60.485 65.749 1.00 14.36 A
ATOM 342 N TYR A 197 69.511 62.011 65.179 1.00 12.60 A
ATOM 343 CA TYR A 197 69.977 62.169 66.556 1.00 13.57 A
ATOM 344 CB TYR A 197 71.221 63.060 66.590 1.00 15.15 A
ATOM 345 CG TYR A 197 71.881 63.190 67.946 1.00 18.18 A
ATOM 346 CD1 TYR A 197 72.287 62.070 68.656 1.00 20.05 A
ATOM 347 CE1 TYR A 197 72.937 62.193 69.878 1.00 21.76 A
ATOM 348 CD2 TYR A 197 72.137 64.440 68.491 1.00 21.37 A
ATOM 349 CE2 TYR A 197 72.784 64.573 69.704 1.00 23.24 A
ATOM 350 CZ TYR A 197 73.181 63.449 70.391 1.00 23.94 A
ATOM 351 OH TYR A 197 73.832 63.589 71.597 1.00 26.71 A
ATOM 352 C TYR A 197 68.852 62.785 67.389 1.00 13.72 A
ATOM 353 0 TYR A 197 68.581 62.343 68.504 1.00 16.14 A
ATOM 354 N ALA A 198 68.194 63.801 66.835 1.00 14.59 A
ATOM 355 CA ALA A 198 67.086 64.466 67.526 1.00 15.09 A
ATOM 356 CB ALA A 198 66.577 65.638 66.687 1.00 14.63 A
ATOM 357 C ALA A 198 65.947 63.484 67.802 1.00 15.02 A
ATOM 358 0 ALA A 198 65.384 63,.454 68.905 1.00 16.63 A
ATOM 359 N ILE A 199 65.604 62.684 66.798 1.00 14.17 A
ATOM 360 CA ILE A 199 64.536 61.699 66.931 1.00 14.10 A
ATOM 361 CB ILE A 199 64.274 60.980 65.585 1.00 15.41 A
ATOM 362 CG2 ILE A 199 63.413 59.741 65.808 1.00 15.83 A
ATOM 363 CG1 ILE A 199 63.595 61.939 64.606 1.00 14.55 A
ATOM 364 CD1 ILE A 199 63.509 61.406 63.190 1.00 16.04 A
ATOM 365 C ILE A 199 64.886 60.660 67.985 1.00 14.55 A
ATOM 366 0 ILE A 199 64.077 60.351 68.856 1.00 14.78 A
ATOM 367 N PHE A 200 66.100 60.127 67.914 1.00 15.53 A
ATOM 368 CA PHE A 200 66.518 59.121 68.878 1.00 16.28 A
ATOM 369 CB PHE A 200 67.865 58.523 68.475 1.00 16.28 A
ATOM 370 CG PHE A 200 67.737 57.332 67.566 1.00 15.69 A
ATOM 371 CD1 PHE A 200 67.066 57.441 66.358 1.00 16.24 A
ATOM 372 CD2 PHE A 200 68.259 56.098 67.929 1.00 17.07 A
ATOM 373 CE1 PHE A 200 66.916 56.343 65.531 1.00 17.37 A
ATOM 374 CE2 PHE A 200 68.112 54.998 67.105 1.00 16.68 A
ATOM 375 CZ PHE A 200 67.439 55.120 65.903 1.00 17.14 A
ATOM 376 C PHE A 200 66.565 59.654 70.305 1.00 17.38 A
ATOM 377 0 PHE A 200 66.274 58.918 71.249 1.00 19.01 A
ATOM 378 N GLN A 201 66.927 60.923 70.469 1.00 17.67 A
ATOM 379 CA GLN A 201 66.955 61.512 71.805 1.00 18.69 A
ATOM 380 CB GLN A 201 67.711 62.845 71.810 1.00 19.36 A
ATOM 381 CG GLN A 201 69.223 62.701 71.788 1.00 22.43 A
ATOM 382 CD GLN A 201 69.925 63.945 72.299 1.00 27.24 A
ATOM 383 OE1 GLN A 201 71.139 63.946 72.512 1.00 30.40 A
ATOM 384 NE2 GLN A 201 69.162 65.014 72.501 1.00 28.68 A
ATOM 385 C GLN A 201 65.521 61.733 72.282 1.00 18.52 A
ATOM 386 0 GLN A 201 65.193 61.460 73.433 1.00 19.29 A
ATOM 387 N GLU A 202 64.670 62.211 71.378 1.00 18.30 A
ATOM 388 CA GLU A 202 63.263 62.473 71.680 1.00 18.52 A
ATOM 389 CB GLU A 202 62.572 63.031 70.432 1.00 19.84 A
ATOM 390 CG GLU A 202 61.058 63.185 70.517 1.00 22.00 A

ATOM 391 CD GLU A 202 60.620 64.238 71.514 1.00 25.77 A
ATOM 392 OE1 GLU A 202 61.382 65.203 71.735 1.00 27.93 A
ATOM 393 OE2 GLU A 202 59.502 64.107 72.063 1.00 27.98 A
ATOM 394 C GLU A 202 62.531 61.219 72.155 1.00 18.79 A
ATOM 395 0 GLU A 202 61.699 61.285 73.061 1.00 20.57 A
ATOM 396 N ARG A 203 62.850 60.080 71.546 1.00 17.76 A
ATOM 397 CA ARG A 203 62.215 58.810 71.888 1.00 17.84 A
ATOM 398 CB ARG A 203 61.961 57.990 70.619 1.00 17.24 A
ATOM 399 CG ARG A 203 60.875 58.568 69.718 1.00 16.56 A
ATOM 400 CD ARG A 203 60.683 57.741 68.456 1.00 16.79 A
ATOM 401 NE ARG A 203 59.524 58.200 67.691 1.00 15.23 A
ATOM 402 CZ ARG A 203 58.259 57.935 68.009 1.00 17.65 A
ATOM 403 NH1 ARG A 203 57.974 57.201 69.078 1.00 16.96 A
ATOM 404 NH2 ARG A 203 57.275 58.417 67.262 1.00 16.00 A
ATOM 405 C ARG A 203 63.019 57.971 72.875 1.00 17.96 A
ATOM 406 0 ARG A 203 62.615 56.861 73.218 1.00 18.80 A
ATOM 407 N ASP A 204 64.155 58.499 73.319 1.00 18.40 A
ATOM 408 CA ASP A 204 65.011 57.792 74.270 1.00 18.83 A
ATOM 409 CB ASP A 204 64.275 57.597 75.599 1.00 21.85 A
ATOM 410 CG ASP A 204 64.028 58.903 76.325 1.00 26.06 A
ATOM 411 OD1 ASP A 204 65.015 59.614 76.615 1.00 28.65 A
ATOM 412 OD2 ASP A 204 62.851 59.218 76.610 1.00 29.13 A
ATOM 413 C ASP A 204 65.474 56.437 73.748 1.00 17.57 A
ATOM 414 0 ASP A 204 65.700 55.507 74.521 1.00 18.95 A
ATOM 415 N LEU A 205 65.628 56.323 72.433 1.00 16.01 A
ATOM 416 CA LEU A 205 66.060 55.064 71.843 1.00 16.61 A
ATOM 417 CB LEU A 205 65.874 55.103 70.324 1.00 17.33 A
ATOM 418 CG LEU A 205 64.421 55.006 69.843 1.00 19.09 A
ATOM 419 CD1 LEU A 205 64.346 55.298 68.346 1.00 17.19 A
ATOM 420 CD2 LEU A 205 63.877 53.617 70.141 1.00 18.49 A
ATOM 421 C LEU A 205 67.506 54.723 72.188 1.00 16.22 A
ATOM 422 0 LEU A 205 67.863 53.548 72.287 1.00 17.42 A
ATOM 423 N LEU A 206 68.338 55.745 72.373 1.00 17.08 A
ATOM 424 CA LEU A 206 69.739 55.510 72.717 1.00 17.81 A
ATOM 425 CB LEU A 206 70.525 56.826 72.711 1.00 18.79 A
ATOM 426 CG LEU A 206 70.599 57.603 71.394 1.00 19.14 A
ATOM 427 CD1 LEU A 206 71.435 58.856 71.591 1.00 20.05 A
ATOM 428 CD2 LEU A 206 71.207 56.726 70.308 1.00 20.42 A
ATOM 429 C LEU A 206 69.825 54.865 74.098 1.00 18.22 A
ATOM 430 0 LEU A 206 70.646 53.980 74.331 1.00 18.94 A
ATOM 431 N LYS A 207 68.966 55.306 75.012 1.00 18.45 A
ATOM 432 CA LYS A 207 68.953 54.757 76.364 1.00 18.71 A
ATOM 433 CB LYS A 207 68.138 55.653 77.296 1.00 19.50 A
ATOM 434 CG LYS A 207 68.716 57.040 77.483 1.00 22.32 A
ATOM 435 CD LYS A 207 67.788 57.909 78.313 1.00 25.07 A
ATOM 436 CE LYS A 207 67.516 57.281 79.671 1.00 25.35 A
ATOM 437 NZ LYS A 207 68.773 57.077 80.444 1.00 28.80 A
ATOM 438 C LYS A 207 68.360 53.355 76.382 1.00 18.50 A
ATOM 439 0 LYS A 207 68.910 52.444 76.999 1.00 20.56 A
ATOM 440 N THR A 208 67.234 53.190 75.697 1.00 17.44 A
ATOM 441 CA THR A 208 66.542 51.911 75.641 1.00 17.30 A
ATOM 442 CB THR A 208 65.281 52.018 74.767 1.00 18.22 A
ATOM 443 001 THR A 208 64.394 52.993 75.329 1.00 21.21 A
ATOM 444 CG2 THR A 208 64.575 50.677 74.688 1.00 19.32 A
ATOM 445 C THR A 208 67.398 50.770 75.111 1.00 17.28 A
ATOM 446 0 THR A 208 67.307 49.635 75.588 1.00 18.99 A
ATOM 447 N PHE A 209 68.236 51.069 74.127 1.00 16.89 A
ATOM 448 CA PHE A 209 69.074 50.044 73.527 1.00 17.34 A
ATOM 449 CB PHE A 209 68.799 50.001 72.020 1.00 15.74 A
ATOM 450 CG PHE A 209 67.400 49.568 71.686 1.00 15.73 A
ATOM 451 CD1 PHE A 209 67.016 48.246 71.851 1.00 16.26 A
ATOM 452 CD2 PHE A 209 66.456 50.488 71.253 1.00 14.45 A
ATOM 453 CE1 PHE A 209 65.715 47.845 71.592 1.00 16.69 A
ATOM 454 CE2 PHE A 209 65.150 50.095 70.991 1.00 15.08 A
ATOM 455 CZ PHE A 209 64.779 48.773 71.161 1.00 16.27 A
ATOM 456 C PHE A 209 70.563 50.227 73.811 1.00 17.36 A
ATOM 457 0 PHE A 209 71.407 49.653 73.123 1.00 18.33 A

ATOM 458 N ARG A 210 70.873 51.018 74.836 1.00 17.44 A
ATOM 459 CA ARG A 210 72.255 51.279 75.234 1.00 17.78 A
ATOM 460 CB ARG A 210 72.816 50.085 76.015 1.00 19.77 A
ATOM 461 CG ARG A 210 72.240 49.913 77.422 1.00 21.38 A
ATOM 462 CD ARG A 210 72.813 50.926 78.405 1.00 21.06 A
ATOM 463 NE ARG A 210 72.315 50.709 79.763 1.00 22.04 A
ATOM 464 CZ ARG A 210 72.738 51.373 80.835 1.00 21.09 A
ATOM 465 NH1 ARG A 210 73.676 52.306 80.720 1.00 20.83 A
ATOM 466 NH2 ARG A 210 72.222 51.105 82.028 1.00 23.00 A
ATOM 467 C ARG A 210 73.171 51.592 74.057 1.00 17.17 A
ATOM 468 0 ARG A 210 74.240 50.995 73.904 1.00 17.95 A
ATOM 469 N ILE A 211 72.751 52.533 73.223 1.00 16.87 A
ATOM 470 CA ILE A 211 73.558 52.924 72.078 1.00 15.09 A
ATOM 471 CB ILE A 211 72.683 53.313 70.867 1.00 16.27 A
ATOM 472 CG2 ILE A 211 73.570 53.588 69.663 1.00 16.35 A
ATOM 473 CG1 ILE A 211 71.685 52.201 70.560 1.00 15.77 A
ATOM 474 CD1 ILE A 211 70.651 52.584 69.501 1.00 13.41 A
ATOM 475 C ILE A 211 74.350 54.148 72.488 1.00 16.35 A
ATOM 476 0 ILE A 211 73.769 55.157 72.885 1.00 17.22 A
ATOM 477 N SER A 212 75.673 54.063 72.413 1.00 15.50 A
ATOM 478 CA SER A 212 76.496 55.205 72.769 1.00 15.44 A
ATOM 479 CB SER A 212 77.963 54.790 72.866 1.00 17.53 A
ATOM 480 OG SER A 212 78.147 53.929 73.976 1.00 17.99 A
ATOM 481 C SER A 212 76.315 56.287 71.710 1.00 16.65 A
ATOM 482 0 SER A 212 76.189 55.992 70.520 1.00 15.73 A
ATOM 483 N SER A 213 76.287 57.539 72.145 1.00 17.02 A
ATOM 484 CA SER A 213 76.105 58.652 71.226 1.00 18.62 A
ATOM 485 CB SER A 213 76.047 59.971 71.997 1.00 20.29 A
ATOM 486 OG SER A 213 74.867 60.041 72.778 1.00 26.41 A
ATOM 487 C SER A 213 77.191 58.729 70.161 1.00 17.78 A
ATOM 488 0 SER A 213 76.901 59.044 69.002 1.00 18.48 A
ATOM 489 N ASP A 214 78.438 58.453 70.534 1.00 17.39 A
ATOM 490 CA ASP A 214 79.500 58.520 69.544 1.00 16.63 A
ATOM 491 CB ASP A 214 80.897 58.424 70.190 1.00 17.62 A
ATOM 492 CG ASP A 214 81.013 57.330 71.239 1.00 18.25 A
ATOM 493 OD1 ASP A 214 80.167 56.413 71.280 1.00 20.25 A
ATOM 494 OD2 ASP A 214 81.990 57.390 72.022 1.00 21.51 A
ATOM 495 C ASP A 214 79.320 57.455 68.470 1.00 15.62 A
ATOM 496 0 ASP A 214 79.527 57.725 67.288 1.00 16.48 A
ATOM 497 N THR A 215 78.923 56.253 68.876 1.00 14.14 A
ATOM 498 CA THR A 215 78.694 55.165 67.931 1.00 14.08 A
ATOM 499 CB THR A 215 78.267 53.882 68.662 1.00 16.27 A
ATOM 500 OG1 THR A 215 79.252 53.551 69.650 1.00 17.56 A
ATOM 501 CG2 THR A 215 78.129 52.729 67.676 1.00 16.78 A
ATOM 502 C THR A 215 77.575 55.566 66.978 1.00 12.71 A
ATOM 503 0 THR A 215 77.692 55.430 65.753 1.00 13.36 A
ATOM 504 N PHE A 216 76.485 56.068 67.548 1.00 12.54 A
ATOM 505 CA PHE A 216 75.334 56.474 66.750 1.00 11.93 A
ATOM 506 CB PHE A 216 74.229 57.026 67.657 1.00 11.50 A
ATOM 507 CG PHE A 216 72.953 57.335 66.928 1.00 13.94 A
ATOM 508 CD1 PHE A 216 72.127 56.314 66.500 1.00 15.73 A
ATOM 509 CD2 PHE A 216 72.596 58.640 66.654 1.00 14.80 A
ATOM 510 CE1 PHE A 216 70.958 56.590 65.803 1.00 19.14 A
ATOM 511 CE2 PHE A 216 71.430 58.924 65.959 1.00 17.38 A
ATOM 512 CZ PHE A 216 70.613 57.896 65.535 1.00 15.37 A
ATOM 513 C PHE A 216 75.702 57.529 65.717 1.00 11.99 A
ATOM 514 0 PHE A 216 75.375 57.398 64.536 1.00 12.76 A
ATOM 515 N ILE A 217 76.388 58.577 66.159 1.00 13.73 A
ATOM 516 CA ILE A 217 76.769 59.645 65.251 1.00 14.45 A
ATOM 517 CB ILE A 217 77.380 60.837 66.017 1.00 16.01 A
ATOM 518 CG2 ILE A 217 77.907 61.877 65.036 1.00 17.64 A
ATOM 519 CG1 ILE A 217 76.312 61.464 66.917 1.00 17.42 A
ATOM 520 CD1 ILE A 217 76.810 62.595 67.780 1.00 20.00 A
ATOM 521 C ILE A 217 77.735 59.153 64.179 1.00 12.59 A
ATOM 522 0 ILE A 217 77.615 59.529 63.014 1.00 14.27 A
ATOM 523 N THR A 218 78.685 58.302 64.556 1.00 12.06 A
ATOM 524 CA THR A 218 79.627 57.791 63.569 1.00 11.98 A

ATOM 525 CB THR A 218 80.711 56.909 64.221 1.00 12.77 A
ATOM 526 OG1 THR A 218 81.393 57.666 65.232 1.00 15.65 A
ATOM 527 CG2 THR A 218 81.725 56.456 63.184 1.00 11.72 A
ATOM 528 C THR A 218 78.875 56.986 62.503 1.00 11.22 A
ATOM 529 0 THR A 218 79.144 57.129 61.315 1.00 12.99 A
ATOM 530 N TYR A 219 77.925 56.154 62.918 1.00 11.49 A
ATOM 531 CA TYR A 219 77.168 55.379 61.936 1.00 10.29 A
ATOM 532 CB TYR A 219 76.205 54.382 62.593 1.00 11.04 A
ATOM 533 CG TYR A 219 75.303 53.713 61.568 1.00 12.11 A
ATOM 534 CD1 TYR A 219 75.751 52.638 60.819 1.00 12.70 A
ATOM 535 CE1 TYR A 219 74.979 52.080 59.820 1.00 10.61 A
ATOM 536 CD2 TYR A 219 74.040 54.216 61.291 1.00 12.48 A
ATOM 537 CE2 TYR A 219 73.260 53.670 60.287 1.00 13.01 A
ATOM 538 CZ TYR A 219 73.738 52.602 59.557 1.00 10.75 A
ATOM 539 OH TYR A 219 72.981 52.054 58.548 1.00 13.24 A
ATOM 540 C TYR A 219 76.350 56.294 61.038 1.00 10.64 A
ATOM 541 0 TYR A 219 76.395 56.167 59.817 1.00 11.75 A
ATOM 542 N MET A 220 75.608 57.218 61.643 1.00 10.51 A
ATOM 543 CA MET A 220 74.766 58.118 60.859 1.00 11.01 A
ATOM 544 CB MET A 220 73.922 59.004 61.778 1.00 12.90 A
ATOM 545 CG MET A 220 72.818 58.250 62.513 1.00 14.37 A
ATOM 546 SD MET A 220 71.716 57.345 61.396 1.00 14.85 A
ATOM 547 CE MET A 220 71.340 58.626 60.167 1.00 16.89 A
ATOM 548 C MET A 220 75.550 58.978 59.882 1.00 11.47 A
ATOM 549 0 MET A 220 75.116 59.177 58.746 1.00 12.12 A
ATOM 550 N MET A 221 76.704 59.485 60.312 1.00 12.71 A
ATOM 551 CA MET A 221 77.538 60.307 59.441 1.00 12.31 A
ATOM 552 CB MET A 221 78.717 60.891 60.229 1.00 11.59 A
ATOM 553 CG MET A 221 78.327 62.007 61.177 1.00 14.51 A
ATOM 554 SD MET A 221 77.825 63.499 60.295 1.00 18.63 A
ATOM 555 CE MET A 221 77.867 64.734 61.640 1.00 19.55 A
ATOM 556 C MET A 221 78.058 59.463 58.285 1.00 12.18 A
ATOM 557 0 MET A 221 78.195 59.950 57.161 1.00 13.98 A
ATOM 558 N THR A 222 78.345 58.196 58.567 1.00 11.39 A
ATOM 559 CA THR A 222 78.845 57.280 57.542 1.00 11.46 A
ATOM 560 CB THR A 222 79.353 55.984 58.195 1.00 11.09 A
ATOM 561 OG1 THR A 222 80.460 56.301 59.050 1.00 12.56 A
ATOM 562 CG2 THR A 222 79.794 54.979 57.142 1.00 13.19 A
ATOM 563 C THR A 222 77.721 56.976 56.553 1.00 11.41 A
ATOM 564 0 THR A 222 77.924 56.992 55.338 1.00 12.33 A
ATOM 565 N LEU A 223 76.529 56.730 57.083 1.00 12.04 A
ATOM 566 CA LEU A 223 75.361 56.438 56.255 1.00 11.51 A
ATOM 567 CB LEU A 223 74.159 56.111 57.147 1.00 11.24 A
ATOM 568 CG LEU A 223 72.842 55.825 56.417 1.00 14.00 A
ATOM 569 CD1 LEU A 223 72.962 54.525 55.643 1.00 13.15 A
ATOM 570 CD2 LEU A 223 71.706 55.725 57.429 1.00 14.69 A
ATOM 571 C LEU A 223 75.028 57.647 55.385 1.00 11.28 A
ATOM 572 0 LEU A 223 74.789 57.514 54.183 1.00 11.76 A
ATOM 573 N GLU A 224 75.016 58.831 55.993 1.00 12.39 A
ATOM 574 CA GLU A 224 74.692 60.046 55.252 1.00 13.02 A
ATOM 575 CB GLU A 224 74.654 61.253 56.197 1.00 16.60 A
ATOM 576 CG GLU A 224 74.243 62.561 55.527 1.00 19.20 A
ATOM 577 CD GLU A 224 73.410 63.458 56.431 1.00 19.14 A
ATOM 578 OE1 GLU A 224 73.662 63.486 57.654 1.00 19.02 A
ATOM 579 OE2 GLU A 224 72.508 64.153 55.915 1.00 17.18 A
ATOM 580 C GLU A 224 75.677 60.281 54.108 1.00 12.19 A
ATOM 581 0 GLU A 224 75.289 60.755 53.035 1.00 12.42 A
ATOM 582 N ASP A 225 76.941 59.931 54.327 1.00 11.04 A
ATOM 583 CA ASP A 225 77.967 60.090 53.302 1.00 14.95 A
ATOM 584 CB ASP A 225 79.363 59.868 53.884 1.00 19.81 A
ATOM 585 CG ASP A 225 80.041 61.159 54.273 1.00 25.16 A
ATOM 586 OD1 ASP A 225 79.746 62.198 53.643 1.00 28.81 A
ATOM 587 OD2 ASP A 225 80.886 61.135 55.192 1.00 25.73 A
ATOM 588 C ASP A 225 77.779 59.137 52.133 1.00 13.03 A
ATOM 589 0 ASP A 225 78.395 59.310 51.081 1.00 16.08 A
ATOM 590 N HIS A 226 76.947 58.118 52.316 1.00 12.65 A
ATOM 591 CA HIS A 226 76.710 57.165 51.250 1.00 12.26 A

ATOM 592 CB HIS A 226 76.582 55.748 51.807 1.00 14.14 A
ATOM 593 CG HIS A 226 77.902 55.110 52.111 1.00 16.55 A
ATOM 594 CD2 HIS A 226 78.603 54.155 51.455 1.00 16.59 A
ATOM 595 ND1 HIS A 226 78.686 55.496 53.177 1.00 17.29 A
ATOM 596 CE1 HIS A 226 79.815 54.808 53.163 1.00 18.97 A
ATOM 597 NE2 HIS A 226 79.790 53.988 52.128 1.00 17.99 A
ATOM 598 C HIS A 226 75.521 57.536 50.380 1.00 12.17 A
ATOM 599 0 HIS A 226 75.175 56.811 49.453 1.00 12.03 A
ATOM 600 N TYR A 227 74.885 58.660 50.701 1.00 11.39 A
ATOM 601 CA TYR A 227 73.812 59.172 49.861 1.00 11.26 A
ATOM 602 CB TYR A 227 72.777 59.980 50.656 1.00 12.86 A
ATOM 603 CG TYR A 227 71.664 59.146 51.251 1.00 9.91 A
ATOM 604 CD1 TYR A 227 71.803 58.540 52.494 1.00 10.75 A
ATOM 605 CE1 TYR A 227 70.802 57.748 53.016 1.00 8.76 A
ATOM 606 CD2 TYR A 227 70.482 58.930 50.549 1.00 10.92 A
ATOM 607 CE2 TYR A 227 69.477 58.133 51.060 1.00 9.51 A
ATOM 608 CZ TYR A 227 69.641 57.543 52.297 1.00 7.54 A
ATOM 609 OH TYR A 227 68.643 56.746 52.805 1.00 9.94 A
ATOM 610 C TYR A 227 74.594 60.104 48.939 1.00 12.03 A
ATOM 611 0 TYR A 227 75.581 60.707 49.358 1.00 15.36 A
ATOM 612 N HIS A 228 74.174 60.205 47.685 1.00 13.80 A
ATOM 613 CA HIS A 228 74.853 61.061 46.721 1.00 15.41 A
ATOM 614 CB HIS A 228 74.696 60.473 45.321 1.00 16.36 A
ATOM 615 CG HIS A 228 75.417 59.178 45.132 1.00 20.91 A
ATOM 616 CD2 HIS A 228 74.962 57.904 45.072 1.00 22.59 A
ATOM 617 ND1 HIS A 228 76.788 59.102 45.002 1.00 24.71 A
ATOM 618 CE1 HIS A 228 77.146 57.837 44.869 1.00 24.66 A
ATOM 619 NE2 HIS A 228 76.056 57.090 44.908 1.00 24.98 A
ATOM 620 C HIS A 228 74.288 62.476 46.753 1.00 16.73 A
ATOM 621 0 HIS A 228 73.129 62.700 46.410 1.00 18.52 A
ATOM 622 N SER A 229 75.111 63.441 47.141 1.00 18.38 A
ATOM 623 CA SER A 229 74.640 64.818 47.217 1.00 19.25 A
ATOM 624 CB SER A 229 75.646 65.677 47.984 1.00 21.64 A
ATOM 625 OG SER A 229 76.889 65.726 47.311 1.00 24.89 A
ATOM 626 C SER A 229 74.361 65.448 45.855 1.00 18.22 A
ATOM 627 0 SER A 229 73.638 66.440 45.772 1.00 20.39 A
ATOM 628 N ASP A 230 74.918 64.869 44.793 1.00 17.49 A
ATOM 629 CA ASP A 230 74.721 65.392 43.443 1.00 18.82 A
ATOM 630 CB ASP A 230 75.918 65.044 42.555 1.00 22.61 A
ATOM 631 CG ASP A 230 76.154 63.552 42.462 1.00 24.85 A
ATOM 632 OD1 ASP A 230 76.454 62.934 43.506 1.00 28.07 A
ATOM 633 OD2 ASP A 230 76.039 62.998 41.348 1.00 30.23 A
ATOM 634 C ASP A 230 73.438 64.888 42.787 1.00 18.02 A
ATOM 635 0 ASP A 230 73.090 65.314 41.683 1.00 19.13 A
ATOM 636 N VAL A 231 72.742 63.970 43.451 1.00 16.44 A
ATOM 637 CA VAL A 231 71.474 63.471 42.922 1.00 15.58 A
ATOM 638 CB VAL A 231 71.159 62.059 43.467 1.00 16.63 A
ATOM 639 CG1 VAL A 231 69.745 61.646 43.094 1.00 18.43 A
ATOM 640 CG2 VAL A 231 72.158 61.062 42.890 1.00 17.55 A
ATOM 641 C VAL A 231 70.453 64.495 43.413 1.00 14.08 A
ATOM 642 0 VAL A 231 70.442 64.845 44.586 1.00 15.21 A
ATOM 643 N ALA A 232 69.601 64.978 42.517 1.00 13.51 A
ATOM 644 CA ALA A 232 68.643 66.020 42.871 1.00 12.81 A
ATOM 645 CB ALA A 232 67.901 66.476 41.619 1.00 14.69 A
ATOM 646 C ALA A 232 67.637 65.737 43.974 1.00 11.78 A
ATOM 647 0 ALA A 232 67.407 66.584 44.839 1.00 14.83 A
ATOM 648 N TYR A 233 67.024 64.565 43.947 1.00 10.72 A
ATOM 649 CA TYR A 233 66.010 64.254 44.946 1.00 8.52 A
ATOM 650 CB TYR A 233 64.720 63.832 44.241 1.00 10.59 A
ATOM 651 CG TYR A 233 63.578 63.475 45.165 1.00 11.48 A
ATOM 652 CD1 TYR A 233 62.720 64.452 45.656 1.00 13.78 A
ATOM 653 CE1 TYR A 233 61.646 64.119 46.467 1.00 11.86 A
ATOM 654 CD2 TYR A 233 63.336 62.156 45.517 1.00 10.75 A
ATOM 655 CE2 TYR A 233 62.272 61.812 46.324 1.00 11.05 A
ATOM 656 CZ TYR A 233 61.425 62.796 46.794 1.00 9.98 A
ATOM 657 OH TYR A 233 60.333 62.438 47.555 1.00 10.85 A
ATOM 658 C TYR A 233 66.417 63.186 45.942 1.00 10.01 A

ATOM 659 0 TYR A 233 66.295 63.384 47.147 1.00 11.44 A
ATOM 660 N HIS A 234 66.900 62.058 45.437 1.00 10.22 A
ATOM 661 CA HIS A 234 67.281 60.962 46.312 1.00 10.33 A
ATOM 662 CB HIS A 234 67.209 59.639 45.547 1.00 11.41 A
ATOM 663 CG HIS A 234 65.811 59.245 45.176 1.00 9.67 A
ATOM 664 CD2 HIS A 234 64.793 58.773 45.934 1.00 10.89 A
ATOM 665 ND1 HIS A 234 65.317 59.348 43.893 1.00 12.98 A
ATOM 666 CE1 HIS A 234 64.056 58.953 43.877 1.00 11.92 A
ATOM 667 NE2 HIS A 234 63.713 58.600 45.102 1.00 12.52 A
ATOM 668 C HIS A 234 68.619 61.116 47.015 1.00 11.24 A
ATOM 669 0 HIS A 234 69.513 60.281 46.883 1.00 13.22 A
ATOM 670 N ASN A 235 68.735 62,197 47.776 1.00 11.39 A
ATOM 671 CA ASN A 235 69.936 62.473 48.552 1.00 10.27 A
ATOM 672 CB ASN A 235 70.496 63.855 48.209 1.00 11.41 A
ATOM 673 CG ASN A 235 69.437 64.928 48.228 1.00 12.84 A
ATOM 674 OD1 ASN A 235 68.772 65.133 49.239 1.00 14.78 A
ATOM 675 ND2 ASN A 235 69.268 65.617 47.103 1.00 12.83 A
ATOM 676 C ASN A 235 69.537 62.386 50.025 1.00 10.06 A
ATOM 677 0 ASN A 235 68.392 62.053 50.335 1.00 10.99 A
ATOM 678 N SER A 236 70.462 62.694 50.930 1.00 10.36 A
ATOM 679 CA SER A 236 70.165 62.568 52.357 1.00 9.20 A
ATOM 680 CB SER A 236 71.433 62.785 53.202 1.00 9.86 A
ATOM 681 OG SER A 236 71.790 64.147 53.303 1.00 13.00 A
ATOM 682 C SER A 236 69.031 63.440 52.878 1.00 10.04 A
ATOM 683 0 SER A 236 68.472 63.150 53.930 1.00 10.71 A
ATOM 684 N LEU A 237 68.679 64.495 52.151 1.00 10.84 A
ATOM 685 CA LEU A 237 67.580 65.356 52.589 1.00 11.27 A
ATOM 686 CB LEU A 237 67.501 66.615 51.719 1.00 13.43 A
ATOM 687 CG LEU A 237 66.576 67.705 52.261 1.00 18.43 A
ATOM 688 CD1 LEU A 237 67.091 68.191 53.609 1.00 18.80 A
ATOM 689 CD2 LEU A 237 66.513 68.859 51.272 1.00 19.22 A
ATOM 690 C LEU A 237 66.264 64.578 52.507 1.00 10.32 A
ATOM 691 0 LEU A 237 65.404 64.686 53.382 1.00 11.99 A
ATOM 692 N HIS A 238 66.103 63.799 51.443 1.00 9.70 A
ATOM 693 CA HIS A 238 64.898 62.997 51.302 1.00 9,52 A
ATOM 694 CB HIS A 238 64.879 62.342 49.917 1.00 9.43 A
ATOM 695 CG HIS A 238 63.864 61.250 49.774 1.00 10.25 A
ATOM 696 CD2 HIS A 238 64.018 59.922 49.569 1.00 10.24 A
ATOM 697 ND1 HIS A 238 62.504 61.472 49.845 1.00 9.33 A
ATOM 698 CE1 HIS A 238 61.867 60.326 49.689 1.00 9.88 A
ATOM 699 NE2 HIS A 238 62.762 59.371 49.520 1.00 10.04 A
ATOM 700 C HIS A 238 64.837 61.941 52.412 1.00 9.29 A
ATOM 701 0 HIS A 238 63.782 61.712 53.005 1.00 9.64 A
ATOM 702 N ALA A 239 65.970 61.308 52.707 1.00 9.53 A
ATOM 703 CA ALA A 239 66.008 60.297 53.759 1.00 9.61 A
ATOM 704 CB ALA A 239 67.396 59.666 53.835 1.00 10.48 A
ATOM 705 C ALA A 239 65.644 60.916 55.111 1.00 8.43 A
ATOM 706 0 ALA A 239 64.906 60.317 55.892 1.00 10.38 A
ATOM 707 N ALA A 240 66.172 62.105 55.384 1.00 10.61 A
ATOM 708 CA ALA A 240 65.873 62.787 56.644 1.00 9.33 A
ATOM 709 CB ALA A 240 66.664 64.093 56.738 1.00 11.23 A
ATOM 710 C ALA A 240 64.371 63.069 56.717 1.00 10.00 A
ATOM 711 0 ALA A 240 63.738 62.864 57.746 1.00 11.21 A
ATOM 712 N ASP A 241 63.808 63.539 55.608 1.00 9.95 A
ATOM 713 CA ASP A 241 62.388 63.837 55.532 1.00 10.00 A
ATOM 714 CB ASP A 241 62.065 64.407 54.151 1.00 11.71 A
ATOM 715 CG ASP A 241 60.588 64.699 53.969 1.00 14.13 A
ATOM 716 OD1 ASP A 241 59.972 65.281 54.888 1.00 16.35 A
ATOM 717 OD2 ASP A 241 60.049 64.351 52.903 1.00 15.16 A
ATOM 718 C ASP A 241 61.541 62.596 55.801 1.00 9.21 A
ATOM 719 0 ASP A 241 60.597 62.639 56.593 1.00 10.80 A
ATOM 720 N VAL A 242 61.884 61.488 55.151 1.00 9.21 A
ATOM 721 CA VAL A 242 61.127 60.260 55.346 1.00 9.40 A
ATOM 722 CB VAL A 242 61.567 59.161 54.343 1.00 8.76 A
ATOM 723 CG1 VAL A 242 60.802 57.869 54.602 1.00 9.67 A
ATOM 724 CG2 VAL A 242 61.277 59.624 52.927 1.00 9.44 A
ATOM 725 C VAL A 242 61.249 59.763 56.790 1.00 9.12 A

ATOM 726 0 VAL A 242 60.271 59.277 57.358 1.00 11.25 A
ATOM 727 N ALA A 243 62.433 59.899 57.386 1.00 11.03 A
ATOM 728 CA ALA A 243 62.636 59.475 58.769 1.00 10.54 A
ATOM 729 CB ALA A 243 64.118 59.564 59.144 1.00 13.23 A
ATOM 730 C ALA A 243 61.806 60.329 59.732 1.00 10.95 A
ATOM 731 0 ALA A 243 61.154 59.806 60.637 1.00 12.73 A
ATOM 732 N GLN A 244 61.836 61.644 59.537 1.00 11.47 A
ATOM 733 CA GLN A 244 61.085 62.551 60.402 1.00 11.53 A
ATOM 734 CB GLN A 244 61.472 64.000 60.102 1.00 11.85 A
ATOM 735 CG GLN A 244 60.948 65.042 61.096 1.00 13.27 A
ATOM 736 CD GLN A 244 59.581 65.569 60.730 1.00 13.24 A
ATOM 737 OE1 GLN A 244 59.236 65.675 59.552 1.00 15.07 A
ATOM 738 NE2 GLN A 244 58.803 65.934 61.739 1.00 12.75 A
ATOM 739 C GLN A 244 59.587 62.346 60.209 1.00 9.52 A
ATOM 740 0 GLN A 244 58.817 62.433 61.162 1.00 12.02 A
ATOM 741 N SER A 245 59.171 62.060 58.978 1.00 10.84 A
ATOM 742 CA SER A 245 57.757 61.828 58.708 1.00 11.18 A
ATOM 743 CB SER A 245 57.502 61.773 57.202 1.00 9.69 A
ATOM 744 OG SER A 245 57.856 63.008 56.601 1.00 12.01 A
ATOM 745 C SER A 245 57.296 60.530 59.366 1.00 10.42 A
ATOM 746 0 SER A 245 56.184 60.455 59.876 1.00 12.31 A
ATOM 747 N THR A 246 58.157 59.513 59.358 1.00 11.70 A
ATOM 748 CA THR A 246 57.828 58.233 59.979 1.00 12.28 A
ATOM 749 CB THR A 246 58.913 57.174 59.650 1.00 12.43 A
ATOM 750 OG1 THR A 246 58.922 56.944 58.238 1.00 12.38 A
ATOM 751 CG2 THR A 246 58.633 55.851 60.358 1.00 12.81 A
ATOM 752 C THR A 246 57.726 58.449 61.487 1.00 12.48 A
ATOM 753 0 THR A 246 56.854 57.882 62.154 1.00 12.79 A
ATOM 754 N HIS A 247 58.618 59.285 62.012 1.00 12.34 A
ATOM 755 CA HIS A 247 58.639 59.614 63.432 1.00 12.74 A
ATOM 756 CB HIS A 247 59.770 60.609 63.709 1.00 14.22 A
ATOM 757 CG HIS A 247 59.662 61.298 65.033 1.00 15.48 A
ATOM 758 CD2 HIS A 247 59.492 62.605 65.344 1.00 16.84 A
ATOM 759 ND1 HIS A 247 59.725 60.623 66.233 1.00 15.80 A
ATOM 760 CE1 HIS A 247 59.598 61.485 67.226 1.00 17.07 A
ATOM 761 NE2 HIS A 247 59.455 62.694 66.715 1.00 16.99 A
ATOM 762 C HIS A 247 57.291 60.205 63.854 1.00 13.52 A
ATOM 763 0 HIS A 247 56.757 59.869 64.910 1.00 16.35 A
ATOM 764 N VAL A 248 56.734 61.073 63.018 1.00 13.83 A
ATOM 765 CA VAL A 248 55.447 61.685 63.326 1.00 14.65 A
ATOM 766 CB VAL A 248 55.154 62.883 62.397 1.00 15.01 A
ATOM 767 CG1 VAL A 248 53.755 63.416 62.670 1.00 14.97 A
ATOM 768 CG2 VAL A 248 56.172 63.980 62.622 1.00 15.35 A
ATOM 769 C VAL A 248 54.306 60.674 63.204 1.00 15.22 A
ATOM 770 0 VAL A 248 53.427 60.619 64.068 1.00 16.97 A
ATOM 771 N LEU A 249 54.316 59.880 62.135 1.00 15.01 A
ATOM 772 CA LEU A 249 53.271 58.882 61.922 1.00 14.68 A
ATOM 773 CB LEU A 249 53.463 58.176 60.573 1.00 15.30 A
ATOM 774 CG LEU A 249 53.177 59.045 59.344 1.00 14.95 A
ATOM 775 CD1 LEU A 249 53.534 58.297 58.065 1.00 16.15 A
ATOM 776 CD2 LEU A 249 51.706 59.434 59.339 1.00 16.99 A
ATOM 777 C LEU A 249 53.231 57.852 63.048 1.00 15.22 A
ATOM 778 0 LEU A 249 52.163 57.355 63.408 1.00 16.74 A
ATOM 779 N LEU A 250 54.390 57.530 63.610 1.00 16.48 A
ATOM 780 CA LEU A 250 54.440 56.560 64.698 1.00 16.69 A
ATOM 781 CB LEU A 250 55.892 56.221 65.042 1.00 16.80 A
ATOM 782 CG LEU A 250 56.590 55.275 64.063 1.00 17.60 A
ATOM 783 CD1 LEU A 250 58.092 55.365 64.239 1.00 17.25 A
ATOM 784 CD2 LEU A 250 56.092 53.853 64.294 1.00 19.47 A
ATOM 785 C LEU A 250 53.727 57.060 65.951 1.00 17.62 A
ATOM 786 0 LEU A 250 53,306 56.261 66.788 1.00 19.32 A
ATOM 787 N SER A 251 53.591 58.376 66.078 1.00 18.59 A
ATOM 788 CA SER A 251 52.937 58.949 67.249 1.00 19.38 A
ATOM 789 CB SER A 251 53.673 60.212 67.695 1,00 20.39 A
ATOM 790 OG SER A 251 54.932 59.888 68.259 1.00 24.83 A
ATOM 791 C SER A 251 51.457 59.266 67.058 1.00 19.49 A
ATOM 792 0 SER A 251 50.855 59.945 67.892 1.00 21.68 A

ATOM 793 N THR A 252 50.865 58.781 65.972 1.00 19.34 A
ATOM 794 CA THR A 252 49.446 59.025 65.737 1.00 20.18 A
ATOM 795 CB THR A 252 48.974 58.393 64.421 1.00 22.38 A
ATOM 796 OGl THR A 252 49.063 56.969 64.513 1.00 26.69 A
ATOM 797 CG2 THR A 252 49.839 58.864 63.273 1.00 20.24 A
ATOM 798 C THR A 252 48.662 58.400 66.884 1.00 19.56 A
ATOM 799 0 THR A 252 48.908 57.253 67.266 1.00 20.24 A
ATOM 800 N PRO A 253 47.706 59.147 67.453 1.00 20.06 A
ATOM 801 CD PRO A 253 47.332 60.525 67.093 1.00 20.20 A
ATOM 802 CA PRO A 253 46.884 58.662 68.567 1.00 19.84 A
ATOM 803 CB PRO A 253 45.778 59.709 68.649 1.00 20.71 A
ATOM 804 CG PRO A 253 46.500 60.954 68.293 1.00 19.86 A
ATOM 805 C PRO A 253 46.332 57.253 68.375 1.00 19.89 A
ATOM 806 0 PRO A 253 46.320 56.450 69.306 1.00 20.48 A
ATOM 807 N ALA A 254 45.879 56.956 67.163 1.00 19.86 A
ATOM 808 CA ALA A 254 45.319 55.648 66.860 1.00 20.86 A
ATOM 809 CB ALA A 254 44.783 55.635 65.439 1.00 21.87 A
ATOM 810 C ALA A 254 46.309 54.502 67.052 1.00 21.49 A
ATOM 811 0 ALA A 254 45.906 53.363 67.264 1.00 22.33 A
ATOM 812 N LEU A 255 47.602 54.800 66.983 1.00 21.67 A
ATOM 813 CA LEU A 255 48.619 53.763 67.134 1.00 22.22 A
ATOM 814 CB LEU A 255 49.622 53.858 65.981 1.00 21.71 A
ATOM 815 CG LEU A 255 49.038 53.721 64.571 1.00 21.23 A
ATOM 816 CD1 LEU A 255 50.117 53.999 63.535 1.00 20.15 A
ATOM 817 CD2 LEU A 255 48.468 52.330 64.384 1.00 21.31 A
ATOM 818 C LEU A 255 49.360 53.836 68.468 1.00 22.79 A
ATOM 819 0 LEU A 255 50.472 53.323 68.597 1.00 23.35 A
ATOM 820 N ASP A 256 48.745 54.462 69.464 1.00 23.64 A
ATOM 821 CA ASP A 256 49.384 54.591 70.767 1.00 24.21 A
ATOM 822 CB ASP A 256 48.577 55.527 71.672 1.00 25.02 A
ATOM 823 CG ASP A 256 49.185 55.662 73.058 1.00 28.18 A
ATOM 824 OD1 ASP A 256 50.372 56.044 73.158 1.00 29.88 A
ATOM 825 OD2 ASP A 256 48.477 55.388 74.051 1.00 30.80 A
ATOM 826 C ASP A 256 49.578 53.253 71.475 1.00 24.00 A
ATOM 827 0 ASP A 256 48.650 52.453 71.578 1.00 24.48 A
ATOM 828 N ALA A 257 50.798 53.026 71.954 1.00 23.88 A
ATOM 829 CA ALA A 257 51.153 51.811 72.681 1.00 24.43 A
ATOM 830 CB ALA A 257 50.301 51.701 73.940 1.00 25.23 A
ATOM 831 C ALA A 257 51.033 50.529 71.865 1.00 24.05 A
ATOM 832 0 ALA A 257 51.139 49.428 72.411 1.00 25.34 A
ATOM 833 N VAL A 258 50.821 50.663 70.561 1.00 21.49 A
ATOM 834 CA VAL A 258 50.685 49.498 69.696 1.00 21.01 A
ATOM 835 CB VAL A 258 49.992 49.877 68.368 1.00 20.43 A
ATOM 836 CGl VAL A 258 50.111 48.739 67.370 1.00 22.15 A
ATOM 837 CG2 VAL A 258 48.523 50.197 68.621 1.00 22.44 A
ATOM 838 C VAL A 258 52.007 48.806 69.366 1.00 19.79 A
ATOM 839 0 VAL A 258 52.084 47.576 69.348 1.00 20.97 A
ATOM 840 N PHE A 259 53.051 49.591 69.118 1.00 19.04 A
ATOM 841 CA PHE A 259 54.342 49.018 68.744 1.00 18.35 A
ATOM 842 CB PHE A 259 54.939 49.823 67.587 1.00 18.36 A
ATOM 843 CG PHE A 259 54.085 49.830 66.352 1.00 17.46 A
ATOM 844 CD1 PHE A 259 53.876 48.668 65.630 1.00 17.34 A
ATOM 845 CD2 PHE A 259 53.477 50.996 65.925 1.00 18.87 A
ATOM 846 CE1 PHE A 259 53.071 48.668 64.500 1.00 17.24 A
ATOM 847 CE2 PHE A 259 52.671 51.006 64.796 1.00 19.53 A
ATOM 848 CZ PHE A 259 52.468 49.839 64.084 1.00 19.84 A
ATOM 849 C PHE A 259 55.371 48.914 69.857 1.00 17.62 A
ATOM 850 0 PHE A 259 55.384 49.719 70.787 1.00 19.41 A
ATOM 851 N THR A 260 56.237 47.912 69.745 1.00 17.89 A
ATOM 852 CA THR A 260 57.303 47.701 70.718 1.00 17.06 A
ATOM 853 CB THR A 260 57.888 46.283 70.623 1.00 19.51 A
ATOM 854 001 THR A 260 58.512 46.115 69.344 1.00 20.56 A
ATOM 855 CG2 THR A 260 56.794 45.238 70.780 1.00 20.22 A
ATOM 856 C THR A 260 58.419 48.683 70.389 1.00 16.31 A
ATOM 857 0 THR A 260 58.455 49.250 69.290 1.00 15.49 A
ATOM 858 N ASP A 261 59.331 48.890 71.328 1.00 15.94 A
ATOM 859 CA ASP A 261 60.432 49.806 71.074 1.00 17.24 A

ATOM 860 CB ASP A 261 61.286 49.996 72.331 1.00 20.48 A
ATOM 861 CG ASP A 261 60.523 50.682 73.452 1.00 24.85 A
ATOM 862 0D1 ASP A 261 59.665 51.539 73.151 1.00 26.79 A
ATOM 863 OD2 ASP A 261 60.790 50.374 74.632 1.00 26.08 A
ATOM 864 C ASP A 261 61.290 49.298 69.913 1.00 16.71 A
ATOM 865 0 ASP A 261 61.841 50.095 69.156 1.00 17.84 A
ATOM 866 N LEU A 262 61.385 47.979 69.759 1.00 16.16 A
ATOM 867 CA LEU A 262 62.175 47.404 68.672 1.00 15.49 A
ATOM 868 CB LEU A 262 62.284 45.879 68.817 1.00 16.17 A
ATOM 869 CG LEU A 262 63.203 45.190 67.798 1.00 17.54 A
ATOM 870 CD1 LEU A 262 64.645 45.594 68.053 1.00 17.57 A
ATOM 871 CD2 LEU A 262 63.051 43.684 67.889 1.00 20.05 A
ATOM 872 C LEU A 262 61.553 47.743 67.316 1.00 15.26 A
ATOM 873 0 LEU A 262 62.263 48.048 66.363 1.00 15.71 A
ATOM 874 N GLU A 263 60.229 47.683 67.233 1.00 15.63 A
ATOM 875 CA GLU A 263 59.532 47.988 65.988 1.00 15.51 A
ATOM 876 CB GLU A 263 58.055 47.580 66.110 1.00 17.38 A
ATOM 877 CG GLU A 263 57.912 46.087 66.432 1.00 19.32 A
ATOM 878 CD GLU A 263 56.487 45.630 66.695 1.00 21.46 A
ATOM 879 OE1 GLU A 263 55.721 46.366 67.349 1.00 21.68 A
ATOM 880 OE2 GLU A 263 56.141 44.511 66.264 1.00 23.34 A
ATOM 881 C GLU A 263 59.690 49.474 65.659 1.00 14.63 A
ATOM 882 0 GLU A 263 59.839 49.848 64.492 1.00 15.41 A
ATOM 883 N ILE A 264 59.664 50.321 66.685 1.00 14.27 A
ATOM 884 CA ILE A 264 59.844 51.758 66.491 1.00 14.47 A
ATOM 885 CB ILE A 264 59.605 52.512 67.822 1.00 15.75 A
ATOM 886 CG2 ILE A 264 60.112 53.944 67.733 1.00 14.96 A
ATOM 887 CG1 ILE A 264 58.112 52.470 68.151 1.00 15.55 A
ATOM 888 CD1 ILE A 264 57.764 52.974 69.534 1.00 17.94 A
ATOM 889 C ILE A 264 61.265 52.002 65.964 1.00 14.06 A
ATOM 890 0 ILE A 264 61.464 52.743 64.993 1.00 14.36 A
ATOM 891 N LEU A 265 62.245 51.366 66.599 1.00 14.04 A
ATOM 892 CA LEU A 265 63.642 51.479 66.183 1.00 13.62 A
ATOM 893 CB LEU A 265 64.525 50.583 67.056 1.00 14.83 A
ATOM 894 CG LEU A 265 66.014 50.518 66.700 1.00 16.23 A
ATOM 895 CD1 LEU A 265 66.685 51.832 67.054 1.00 17.52 A
ATOM 896 CD2 LEU A 265 66.672 49.370 67.466 1.00 15.63 A
ATOM 897 C LEU A 265 63.787 51.039 64.727 1.00 12.86 A
ATOM 898 0 LEU A 265 64.416 51.722 63.916 1.00 14.63 A
ATOM 899 N ALA A 266 63.193 49.895 64.403 1.00 13.09 A
ATOM 900 CA ALA A 266 63.270 49.348 63.050 1.00 12.95 A
ATOM 901 CB ALA A 266 62.581 47.989 62.997 1.00 13.61 A
ATOM 902 C ALA A 266 62.661 50.273 62.007 1.00 12.42 A
ATOM 903 0 ALA A 266 63.234 50.464 60.932 1.00 13.61 A
ATOM 904 N ALA A 267 61.508 50.849 62.328 1.00 12.74 A
ATOM 905 CA ALA A 267 60.822 51.745 61.402 1.00 12.46 A
ATOM 906 CB ALA A 267 59.456 52.143 61.970 1.00 13.39 A
ATOM 907 C ALA A 267 61.637 52.993 61.084 1.00 12.23 A
ATOM 908 0 ALA A 267 61.765 53.376 59.922 1.00 14.13 A
ATOM 909 N ILE A 268 62.190 53.631 62.111 1.00 12.39 A
ATOM 910 CA ILE A 268 62.977 54.835 61.890 1.00 12.16 A
ATOM 911 CB ILE A 268 63.218 55.585 63.220 1.00 13,76 A
ATOM 912 CG2 ILE A 268 64.085 56.814 62.983 1.00 16.04 A
ATOM 913 CG1 ILE A 268 61.865 56.008 63.809 1.00 16.47 A
ATOM 914 CD1 ILE A 268 61.951 56.633 65.197 1.00 17.84 A
ATOM 915 C ILE A 268 64.297 54.505 61.189 1.00 10.99 A
ATOM 916 0 ILE A 268 64.751 55.251 60.321 1.00 12.73 A
ATOM 917 N PHE A 269 64.906 53.382 61.556 1.00 10.98 A
ATOM 918 CA PHE A 269 66.148 52.964 60.915 1.00 11.22 A
ATOM 919 CB PHE A 269 66.684 51.688 61.573 1.00 12.29 A
ATOM 920 CG PHE A 269 67.942 51.163 60.942 1.00 13.65 A
ATOM 921 CD1 PHE A 269 69.153 51.809 61.131 1.00 14.62 A
ATOM 922 CD2 PHE A 269 67.912 50.024 60.154 1.00 14.97 A
ATOM 923 CE1 PHE A 269 70.313 51.326 60.547 1.00 15.18 A
ATOM 924 CE2 PHE A 269 69.065 49.538 59.567 1.00 13.83 A
ATOM 925 CZ PHE A 269 70.266 50.191 59.766 1.00 15.15 A
ATOM 926 C PHE A 269 65.866 52.696 59.436 1.00 10.84 A

ATOM 927 0 PHE A 269 66.619 53.126 58.557 1.00 11.96 A
ATOM 928 N ALA A 270 64.781 51.979 59.164 1.00 10.96 A
ATOM 929 CA ALA A 270 64.416 51.660 57.786 1.00 10.59 A
ATOM 930 CB ALA A 270 63.133 50.837 57.758 1.00 12.22 A
ATOM 931 C ALA A 270 64.245 52.936 56.964 1.00 9.83 A
ATOM 932 0 ALA A 270 64.720 53.019 55.832 1.00 10.53 A
ATOM 933 N ALA A 271 63.565 53.932 57.527 1.00 10.99 A
ATOM 934 CA ALA A 271 63.362 55.184 56.807 1.00 9.90 A
ATOM 935 CB ALA A 271 62.463 56.115 57.623 1.00 13.45 A
ATOM 936 C ALA A 271 64.702 55.857 56.513 1.00 9.51 A
ATOM 937 0 ALA A 271 64.928 56.378 55.420 1.00 10.70 A
ATOM 938 N ALA A 272 65.608 55.836 57.486 1.00 9.89 A
ATOM 939 CA ALA A 272 66.914 56.458 57.302 1.00 8.82 A
ATOM 940 CB ALA A 272 67.688 56.434 58.619 1.00 10.73 A
ATOM 941 C ALA A 272 67.762 55.822 56.193 1.00 8.10 A
ATOM 942 0 ALA A 272 68.461 56.527 55.461 1.00 10.85 A
ATOM 943 N ILE A 273 67.705 54.499 56.067 1.00 8.73 A
ATOM 944 CA ILE A 273 68.515 53.817 55.055 1.00 8.22 A
ATOM 945 CB ILE A 273 69.077 52.450 55.559 1.00 8.56 A
ATOM 946 CG2 ILE A 273 69.778 52.619 56.890 1.00 11.42 A
ATOM 947 CG1 ILE A 273 67.946 51.423 55.677 1.00 10.89 A
ATOM 948 CD1 ILE A 273 68.441 49.992 55.779 1.00 11.69 A
ATOM 949 C ILE A 273 67.775 53.482 53.776 1.00 9.67 A
ATOM 950 0 ILE A 273 68.398 53.016 52.830 1.00 11.43 A
ATOM 951 N HIS A 274 66.469 53.745 53.723 1.00 9.87 A
ATOM 952 CA HIS A 274 65.680 53.317 52.575 1.00 9.09 A
ATOM 953 CB HIS A 274 64.196 53.638 52.800 1.00 9.86 A
ATOM 954 CG HIS A 274 63.757 54.929 52.195 1.00 11.74 A
ATOM 955 CD2 HIS A 274 63.070 55.188 51.057 1.00 9.64 A
ATOM 956 ND1 HIS A 274 64.048 56.152 52.757 1.00 10.74 A
ATOM 957 CE1 HIS A 274 63.561 57.111 51.991 1.00 11.82 A
ATOM 958 NE2 HIS A 274 62.962 56.552 50.952 1.00 7.53 A
ATOM 959 C HIS A 274 66.086 53.692 51.151 1.00 8.60 A
ATOM 960 0 HIS A 274 65.671 53.007 50.216 1.00 9.73 A
ATOM 961 N ASP A 275 66.882 54.746 50.968 1.00 7.82 A
ATOM 962 CA ASP A 275 67.336 55.133 49.619 1.00 7.44 A
ATOM 963 CB ASP A 275 66.679 56.443 49.154 1.00 9.89 A
ATOM 964 CG ASP A 275 65.359 56.223 48.460 1.00 8.90 A
ATOM 965 OD1 ASP A 275 65.178 55.155 47.849 1.00 9.38 A
ATOM 966 OD2 ASP A 275 64.507 57.130 48.504 1.00 8.55 A
ATOM 967 C ASP A 275 68.848 55.332 49.566 1.00 7.74 A
ATOM 968 0 ASP A 275 69.358 55.982 48.660 1.00 9.34 A
ATOM 969 N VAL A 276 69.580 54.771 50.522 1.00 7.75 A
ATOM 970 CA VAL A 276 71.017 55.001 50.532 1.00 8.61 A
ATOM 971 CB VAL A 276 71.661 54.414 51.810 1.00 10.03 A
ATOM 972 CG1 VAL A 276 71.584 52.914 51.787 1.00 11.00 A
ATOM 973 CG2 VAL A 276 73.099 54.910 51.942 1.00 9.13 A
ATOM 974 C VAL A 276 71.737 54.495 49.282 1.00 8.35 A
ATOM 975 0 VAL A 276 71.423 53.435 48.746 1.00 10.58 A
ATOM 976 N ASP A 277 72.686 55.301 48.817 1.00 9.55 A
ATOM 977 CA ASP A 277 73.485 55.013 47.629 1.00 9.67 A
ATOM 978 CB ASP A 277 74.290 53.720 47.812 1.00 12.86 A
ATOM 979 CG ASP A 277 75.454 53.614 46.835 1.00 15.64 A
ATOM 980 OD1 ASP A 277 76.021 54.666 46.464 1.00 17.72 A
ATOM 981 OD2 ASP A 277 75.818 52.483 46.452 1.00 18.03 A
ATOM 982 C ASP A 277 72.625 54.932 46.371 1.00 9.98 A
ATOM 983 0 ASP A 277 72.902 54.157 45.462 1.00 12.40 A
ATOM 984 N HIS A 278 71.576 55.745 46.332 1.00 10.37 A
ATOM 985 CA HIS A 278 70.679 55.792 45.181 1.00 10.87 A
ATOM 986 CE HIS A 278 69.407 56.557 45.564 1.00 10.73 A
ATOM 987 CG HIS A 278 68.266 56.351 44.617 1.00 12.83 A
ATOM 988 CD2 HIS A 278 67.054 55.773 44.796 1.00 12.74 A
ATOM 989 ND1 HIS A 278 68.300 56.772 43.306 1.00 11.18 A
ATOM 990 CE1 HIS A 278 67.158 56.462 42.717 1.00 12.18 A
ATOM 991 NE2 HIS A 278 66.385 55.854 43.599 1.00 12.59 A
ATOM 992 C HIS A 278 71.437 56.527 44.071 1.00 10.98 A
ATOM 993 0 HIS A 278 71.980 57.600 44.301 1.00 11.48 A

ATOM 994 N PRO A 279 71.499 55.945 42.858 1.00 12.29 A
ATOM 995 CD PRO A 279 71.055 54.576 42.541 1.00 13.89 A
ATOM 996 CA PRO A 279 72.199 56.544 41.714 1.00 12.49 A
ATOM 997 CB PRO A 279 72.476 55.341 40.820 1.00 13.95 A
ATOM 998 CG PRO A 279 71.268 54.499 41.033 1.00 14.11 A
ATOM 999 C PRO A 279 71.461 57.654 40.966 1.00 12.54 A
ATOM 1000 0 PRO A 279 72.026 58.279 40.061 1.00 14.01 A
ATOM 1001 N GLY A 280 70.209 57.902 41.335 1.00 12.41 A
ATOM 1002 CA GLY A 280 69.459 58.951 40.669 1.00 13.96 A
ATOM 1003 C GLY A 280 68.738 58.498 39.417 1.00 15.21 A
ATOM 1004 0 GLY A 280 68.302 59.324 38.613 1.00 15.91 A
ATOM 1005 N VAL A 281 68.634 57.187 39.237 1.00 14.73 A
ATOM 1006 CA VAL A 281 67.923 56.619 38.101 1.00 13.66 A
ATOM 1007 CB VAL A 281 68.885 56.032 37.042 1.00 13.41 A
ATOM 1008 CG1 VAL A 281 69.755 57.147 36.458 1.00 14.56 A
ATOM 1009 CG2 VAL A 281 69.738 54.936 37.654 1.00 15.72 A
ATOM 1010 C VAL A 281 67.010 55.522 38.628 1.00 13.42 A
ATOM 1011 0 VAL A 281 67.209 55.006 39.735 1.00 14.31 A
ATOM 1012 N SER A 282 66.015 55.164 37.827 1.00 12.82 A
ATOM 1013 CA SER A 282 65.027 54.158 38.202 1.00 11.61 A
ATOM 1014 CB SER A 282 63.785 54.334 37.340 1.00 14.29 A
ATOM 1015 OG SER A 282 64.083 53.957 36.006 1.00 14.59 A
ATOM 1016 C SER A 282 65.494 52.713 38.058 1.00 9.29 A
ATOM 1017 0 SER A 282 66.514 52.434 37.426 1.00 10.99 A
ATOM 1018 N ASN A 283 64.716 51.790 38.622 1.00 10.33 A
ATOM 1019 CA ASN A 283 65.043 50.376 38.511 1.00 10.12 A
ATOM 1020 CB ASN A 283 64.017 49.513 39.252 1.00 10.66 A
ATOM 1021 CG ASN A 283 64.278 49.452 40.730 1.00 12.46 A
ATOM 1022 OD1 ASN A 283 65.422 49.554 41.168 1.00 14.88 A
ATOM 1023 ND2 ASN A 283 63.221 49.262 41.513 1.00 12.08 A
ATOM 1024 C ASN A 283 65.065 49.969 37.044 1.00 10.47 A
ATOM 1025 0 ASN A 283 65.911 49.190 36.623 1.00 12.66 A
ATOM 1026 N GLN A 284 64.125 50.498 36.269 1.00 12.15 A
ATOM 1027 CA GLN A 284 64.053 50.161 34.853 1.00 11.81 A
ATOM 1028 CB GLN A 284 62.813 50.790 34.214 1.00 15.62 A
ATOM 1029 CG GLN A 284 62.480 50.202 32.855 1.00 22.18 A
ATOM 1030 CD GLN A 284 62.331 48.691 32.910 1.00 24.03 A
ATOM 1031 OE1 GLN A 284 61.589 48.157 33.741 1.00 25.53 A
ATOM 1032 NE2 GLN A 284 63.036 47.994 32.026 1.00 26.67 A
ATOM 1033 C GLN A 284 65.309 50.611 34.114 1.00 12.41 A
ATOM 1034 0 GLN A 284 65.772 49.940 33.196 1.00 13.25 A
ATOM 1035 N PHE A 285 65.870 51.744 34.522 1.00 12.41 A
ATOM 1036 CA PHE A 285 67.077 52.242 33.879 1.00 11.68 A
ATOM 1037 CB PHE A 285 67.435 53.624 34.437 1.00 11.83 A
ATOM 1038 CG PHE A 285 68.660 54.237 33.814 1.00 10.72 A
ATOM 1039 CD1 PHE A 285 69.927 53.892 34.257 1.00 10.38 A
ATOM 1040 CD2 PHE A 285 68.546 55.147 32.772 1.00 11.44 A
ATOM 1041 CE1 PHE A 285 71.062 54.442 33.678 1.00 13.14 A
ATOM 1042 CE2 PHE A 285 69.680 55.703 32.186 1.00 12.75 A
ATOM 1043 CZ PHE A 285 70.938 55.349 32.641 1.00 13.30 A
ATOM 1044 C PHE A 285 68.214 51.253 34.108 1.00 10.36 A
ATOM 1045 0 PHE A 285 68.955 50.918 33.181 1.00 12.82 A
ATOM 1046 N LEU A 286 68.346 50.779 35.345 1.00 11.39 A
ATOM 1047 CA LEU A 286 69.392 49.821 35.696 1.00 12.33 A
ATOM 1048 CB LEU A 286 69.391 49.565 37.204 1.00 14.20 A
ATOM 1049 CG LEU A 286 69.708 50.785 38.066 1.00 15.06 A
ATOM 1050 CD1 LEU A 286 69.510 50.430 39.528 1.00 14.27 A
ATOM 1051 CD2 LEU A 286 71.126 51.248 37.798 1.00 15.73 A
ATOM 1052 C LEU A 286 69.192 48.501 34.963 1.00 12.49 A
ATOM 1053 0 LEU A 286 70.153 47.840 34.574 1.00 14.41 A
ATOM 1054 N ILE A 287 67.938 48.112 34.781 1.00 13.50 A
ATOM 1055 CA ILE A 287 67.635 46.870 34.095 1.00 14.65 A
ATOM 1056 CB ILE A 287 66.141 46.517 34.267 1.00 14.38 A
ATOM 1057 CG2 ILE A 287 65.738 45.406 33.305 1.00 16.15 A
ATOM 1058 CG1 ILE A 287 65.888 46.104 35.720 1.00 14.21 A
ATOM 1059 CD1 ILE A 287 64.425 45.994 36.082 1.00 14.40 A
ATOM 1060 C ILE A 287 67.993 46.986 32.619 1.00 14.47 A

ATOM 1061 0 ILE A 287 68.615 46.085 32.049 1.00 14.94 A
ATOM 1062 N ASN A 288 67.623 48.111 32.012 1.00 13.61 A
ATOM 1063 CA ASN A 288 67.896 48.342 30.597 1.00 15.19 A
ATOM 1064 CB ASN A 288 67.163 49.592 30.114 1.00 15.01 A
ATOM 1065 CG ASN A 288 65.663 49.419 30.125 1.00 19.03 A
ATOM 1066 OD1 ASN A 288 65.159 48.298 30.051 1.00 20.46 A
ATOM 1067 ND2 ASN A 288 64.936 50.530 30.204 1.00 21.63 A
ATOM 1068 C ASN A 288 69.377 48.466 30.265 1.00 14.72 A
ATOM 1069 0 ASN A 288 69.789 48.171 29.141 1.00 18.40 A
ATOM 1070 N THR A 289 70.180 48.903 31.229 1.00 14.21 A
ATOM 1071 CA THR A 289 71.614 49.052 30.991 1.00 13.57 A
ATOM 1072 CB THR A 289 72.180 50.284 31.732 1.00 14.64 A
ATOM 1073 001 THR A 289 71.852 50.201 33.124 1.00 16.00 A
ATOM 1074 CG2 THR A 289 71.608 51.568 31.151 1.00 14.01 A
ATOM 1075 C THR A 289 72.418 47.816 31.398 1.00 13.31 A
ATOM 1076 0 THR A 289 73.646 47.800 31.288 1.00 14.55 A
ATOM 1077 N ASN A 290 71.721 46.773 31.837 1.00 13.87 A
ATOM 1078 CA ASN A 290 72.374 45.541 32.272 1.00 15.12 A
ATOM 1079 CB ASN A 290 73.137 44.881 31.121 1.00 17.91 A
ATOM 1080 CG ASN A 290 72.225 44.433 30.010 1.00 23.51 A
ATOM 1081 OD1 ASN A 290 71.158 43.867 30.259 1.00 27.23 A
ATOM 1082 ND2 ASN A 290 72.641 44.668 28.771 1.00 26.45 A
ATOM 1083 C ASN A 290 73.336 45.810 33.415 1.00 15.07 A
ATOM 1084 0 ASN A 290 74.436 45.258 33.459 1.00 16.56 A
ATOM 1085 N SER A 291 72.909 46.664 34.338 1.00 15.07 A
ATOM 1086 CA SER A 291 73.707 47.006 35.506 1.00 14.92 A
ATOM 1087 CB SER A 291 72.971 48.038 36.361 1.00 17.76 A
ATOM 1088 OG SER A 291 73.583 48.166 37.636 1.00 25.05 A
ATOM 1089 C SER A 291 73.997 45.775 36.353 1.00 13.85 A
ATOM 1090 0 SER A 291 73.175 44.865 36.447 1.00 13.51 A
ATOM 1091 N GLU A 292 75.172 45.758 36.974 1.00 15.67 A
ATOM 1092 CA GLU A 292 75.570 44.645 37.830 1.00 16.78 A
ATOM 1093 CB GLU A 292 76.915 44.951 38.493 1.00 19.65 A
ATOM 1094 CG GLU A 292 77.358 43.921 39.521 1.00 24.88 A
ATOM 1095 CD GLU A 292 78.737 44.217 40.087 1.00 27.13 A
ATOM 1096 OE1 GLU A 292 78.952 45.347 40.575 1.00 29.29 A
ATOM 1097 OE2 GLU A 292 79.606 43.320 40.046 1.00 30.31 A
ATOM 1098 C GLU A 292 74.516 44.389 38.905 1.00 15.63 A
ATOM 1099 0 GLU A 292 74.250 43.239 39.263 1.00 15.23 A
ATOM 1100 N LEU A 293 73.917 45.465 39.411 1.00 15.27 A
ATOM 1101 CA LEU A 293 72.891 45.349 40.446 1.00 14.59 A
ATOM 1102 CB LEU A 293 72.475 46.734 40.947 1.00 16.52 A
ATOM 1103 CG LEU A 293 73.424 47.464 41.899 1.00 19.49 A
ATOM 1104 CD1 LEU A 293 74.779 47.650 41.257 1.00 24.10 A
ATOM 1105 CD2 LEU A 293 72.817 48.805 42.269 1.00 20.66 A
ATOM 1106 C LEU A 293 71.659 44.608 39.940 1.00 13.51 A
ATOM 1107 0 LEU A 293 71.095 43.768 40.639 1.00 14.29 A
ATOM 1108 N ALA A 294 71.240 44.916 38.720 1.00 13.49 A
ATOM 1109 CA ALA A 294 70.067 44.264 38.161 1.00 14.10 A
ATOM 1110 CB ALA A 294 69.683 44.915 36.832 1.00 15.46 A
ATOM 1111 C ALA A 294 70.334 42.775 37.970 1.00 14.31 A
ATOM 1112 0 ALA A 294 69.462 41.937 38.207 1.00 15.96 A
ATOM 1113 N LEU A 295 71.546 42.441 37.544 1.00 14.87 A
ATOM 1114 CA LEU A 295 71.889 41.045 37.337 1.00 15.39 A
ATOM 1115 CB LEU A 295 73.251 40.941 36.644 1.00 16.41 A
ATOM 1116 CG LEU A 295 73.257 41.520 35.225 1.00 20.94 A
ATOM 1117 CD1 LEU A 295 74.638 41.376 34.618 1.00 21.57 A
ATOM 1118 CD2 LEU A 295 72.221 40.801 34.372 1.00 22.86 A
ATOM 1119 C LEU A 295 71.895 40.294 38.668 1.00 16.39 A
ATOM 1120 0 LEU A 295 71.376 39.185 38.765 1.00 18.29 A
ATOM 1121 N MET A 296 72.462 40.912 39.697 1.00 16.33 A
ATOM 1122 CA MET A 296 72.530 40.296 41.020 1.00 17.98 A
ATOM 1123 CB MET A 296 73.257 41.220 41.996 1.00 24.29 A
ATOM 1124 CG MET A 296 74.705 41.495 41.691 1.00 29.41 A
ATOM 1125 SD MET A 296 75.292 42.798 42.788 1.00 31.48 A
ATOM 1126 CE MET A 296 74.627 42.251 44.337 1.00 29.53 A
ATOM 1127 C MET A 296 71.156 39.997 41.616 1.00 15.96 A

ATOM 1128 0 MET A 296 70.931 38.932 42.203 1.00 16.23 A
ATOM 1129 N TYR A 297 70.243 40.950 41.473 1.00 14.24 A
ATOM 1130 CA TYR A 297 68.918 40.809 42.051 1.00 12.86 A
ATOM 1131 CB TYR A 297 68.525 42.131 42.725 1.00 13.61 A
ATOM 1132 CG TYR A 297 69.411 42.453 43.903 1.00 13.90 A
ATOM 1133 CD1 TYR A 297 69.460 41.607 45.002 1.00 13.95 A
ATOM 1134 CE1 TYR A 297 70.315 41.855 46.057 1.00 15.30 A
ATOM 1135 CD2 TYR A 297 70.240 43.567 43.892 1.00 14.98 A
ATOM 1136 CE2 TYR A 297 71.101 43.824 44.947 1.00 18.07 A
ATOM 1137 CZ TYR A 297 71.131 42.962 46.023 1.00 16.45 A
ATOM 1138 OH TYR A 297 71.985 43.209 47.075 1.00 20.50 A
ATOM 1139 C TYR A 297 67.829 40.340 41.095 1.00 11.93 A
ATOM 1140 0 TYR A 297 66.642 40.446 41.392 1.00 12.79 A
ATOM 1141 N ASN A 298 68.244 39.803 39.954 1.00 11.89 A
ATOM 1142 CA ASN A 298 67.305 39.289 38.970 1.00 11.45 A
ATOM 1143 CB ASN A 298 66.748 37.945 39.452 1.00 12.91 A
ATOM 1144 CG ASN A 298 67.845 36.906 39.619 1.00 14.01 A
ATOM 1145 OD1 ASN A 298 68.662 36.721 38.723 1.00 15.20 A
ATOM 1146 ND2 ASN A 298 67.874 36.235 40.767 1.00 14.19 A
ATOM 1147 C ASN A 298 66.186 40.257 38.609 1.00 10.25 A
ATOM 1148 0 ASN A 298 65.016 39.885 38.507 1.00 12.39 A
ATOM 1149 N ASP A 299 66.579 41.512 38.414 1.00 12.68 A
ATOM 1150 CA ASP A 299 65.676 42.585 38.013 1.00 12.14 A
ATOM 1151 CB ASP A 299 65.122 42.307 36.606 1.00 13.25 A
ATOM 1152 CG ASP A 299 66.218 42.165 35.551 1.00 16.79 A
ATOM 1153 OD1 ASP A 299 67.349 42.648 35.768 1.00 16.70 A
ATOM 1154 OD2 ASP A 299 65.936 41.582 34.480 1.00 17.81 A
ATOM 1155 C ASP A 299 64.512 42.875 38.960 1.00 12.75 A
ATOM 1156 0 ASP A 299 63.593 43.609 38.598 1.00 15.43 A
ATOM 1157 N GLU A 300 64.555 42.328 40.170 1.00 13.14 A
ATOM 1158 CA GLU A 300 63.466 42.543 41.131 1.00 14.50 A
ATOM 1159 CB GLU A 300 63.027 41.203 41.722 1.00 16.17 A
ATOM 1160 CG GLU A 300 62.008 41.337 42.845 1.00 21.34 A
ATOM 1161 CD GLU A 300 60.614 41.692 42.349 1.00 27.09 A
ATOM 1162 OE1 GLU A 300 60.483 42.598 41.496 1.00 29.87 A
ATOM 1163 OE2 GLU A 300 59.641 41.069 42.825 1.00 30.80 A
ATOM 1164 C GLU A 300 63.831 43.498 42.268 1.00 13.55 A
ATOM 1165 0 GLU A 300 64.785 43.247 43.006 1.00 15.42 A
ATOM 1166 N SER A 301 63.054 44.574 42.419 1.00 12.92 A
ATOM 1167 CA SER A 301 63.301 45.578 43.465 1.00 11.80 A
ATOM 1168 CB SER A 301 62.759 45.093 44.816 1.00 13.57 A
ATOM 1169 OG SER A 301 61.351 44.933 44.774 1.00 14.63 A
ATOM 1170 C SER A 301 64.804 45.810 43.558 1.00 10.78 A
ATOM 1171 0 SER A 301 65.394 45.807 44.637 1.00 12.22 A
ATOM 1172 N VAL A 302 65.418 46.045 42.407 1.00 11.02 A
ATOM 1173 CA VAL A 302 66.861 46.211 42.354 1.00 10.12 A
ATOM 1174 CB VAL A 302 67.319 46.536 40.926 1.00 11.34 A
ATOM 1175 CG1 VAL A 302 68.837 46.697 40.891 1.00 11.71 A
ATOM 1176 CG2 VAL A 302 66.869 45.422 39.985 1.00 13.02 A
ATOM 1177 C VAL A 302 67.452 47.236 43.308 1.00 9.28 A
ATOM 1178 0 VAL A 302 68.333 46.909 44.108 1.00 12.15 A
ATOM 1179 N LEU A 303 66.980 48.473 43.234 1.00 11.21 A
ATOM 1180 CA LEU A 303 67.523 49.510 44.098 1.00 10.85 A
ATOM 1181 CB LEU A 303 67.016 50.890 43.664 1.00 14.89 A
ATOM 1182 CG LEU A 303 67.774 51.471 42.467 1.00 18.24 A
ATOM 1183 CD1 LEU A 303 67.149 52.779 42.024 1.00 21.91 A
ATOM 1184 CD2 LEU A 303 69.235 51.673 42.861 1.00 17.26 A
ATOM 1185 C LEU A 303 67.209 49.286 45.565 1.00 10.37 A
ATOM 1186 0 LEU A 303 68.065 49.495 46.418 1.00 11.02 A
ATOM 1187 N GLU A 304 65.986 48.856 45.854 1.00 10.15 A
ATOM 1188 CA GLU A 304 65.576 48.620 47.232 1.00 10.99 A
ATOM 1189 CB GLU A 304 64.098 48.222 47.275 1.00 13.64 A
ATOM 1190 CG GLU A 304 63.117 49.329 46.828 1.00 14.71 A
ATOM 1191 CD GLU A 304 63.172 49.652 45.334 1.00 15.64 A
ATOM 1192 OE1 GLU A 304 63.510 48.760 44.526 1.00 13.16 A
ATOM 1193 OE2 GLU A 304 62.847 50.802 44.960 1.00 15.63 A
ATOM 1194 C GLU A 304 66.451 47.546 47.891 1.00 10.98 A

ATOM 1195 0 GLU A 304 66.809 47.662 49.066 1.00 12.37 A
ATOM 1196 N ASN A 305 66.801 46.504 47.144 1.00 10.94 A
ATOM 1197 CA ASN A 305 67.666 45.463 47.697 1.00 10.24 A
ATOM 1198 CB ASN A 305 67.753 44.273 46.736 1.00 11.47 A
ATOM 1199 CG ASN A 305 66.638 43.276 46.959 1.00 13.72 A
ATOM 1200 OD1 ASN A 305 66.591 42.613 47.992 1.00 16.46 A
ATOM 1201 ND2 ASN A 305 65.729 43.173 46.001 1.00 15.40 A
ATOM 1202 C ASN A 305 69.054 46.038 47.950 1.00 9.96 A
ATOM 1203 0 ASN A 305 69.711 45.705 48.938 1.00 12.28 A
ATOM 1204 N HIS A 306 69.509 46.904 47.054 1.00 12.09 A
ATOM 1205 CA HIS A 306 70.812 47.525 47.230 1.00 11.27 A
ATOM 1206 CB HIS A 306 71.189 48.331 45.986 1.00 13.44 A
ATOM 1207 CG HIS A 306 72.535 48,975 46.079 1.00 13.87 A
ATOM 1208 CD2 HIS A 306 72.908 50.276 46.038 1.00 15.76 A
ATOM 1209 ND1 HIS A 306 73.691 48.249 46.270 1.00 15.82 A
ATOM 1210 CE1 HIS A 306 74.718 49.076 46.345 1.00 17.17 A
ATOM 1211 NE2 HIS A 306 74.271 50.312 46.208 1.00 15.59 A
ATOM 1212 C HIS A 306 70.816 48.438 48.459 1.00 10.63 A
ATOM 1213 0 HIS A 306 71.772 48.435 49.229 1.00 10.64 A
ATOM 1214 N HIS A 307 69.745 49.211 48.651 1.00 10.21 A
ATOM 1215 CA HIS A 307 69.676 50.112 49.807 1.00 10.36 A
ATOM 1216 CB HIS A 307 68.350 50.894 49.830 1.00 10.66 A
ATOM 1217 CG HIS A 307 68.074 51.673 48.578 1.00 8.93 A
ATOM 1218 CD2 HIS A 307 66,937 51.816 47.855 1.00 9.54 A
ATOM 1219 ND1 HIS A 307 69.032 52.432 47.941 1.00 10.97 A
ATOM 1220 CE1 HIS A 307 68.498 53.006 46.875 1.00 10.16 A
ATOM 1221 NE2 HIS A 307 67.229 52.648 46.801 1.00 10.25 A
ATOM 1222 C HIS A 307 69.806 49.309 51.101 1.00 9.63 A
ATOM 1223 0 HIS A 307 70.486 49.720 52.040 1.00 10.40 A
ATOM 1224 N LEU A 308 69.138 48.163 51.149 1.00 9.63 A
ATOM 1225 CA LEU A 308 69.205 47.294 52.318 1.00 10.46 A
ATOM 1226 CB LEU A 308 68.249 46.112 52.144 1.00 11.26 A
ATOM 1227 CG LEU A 308 66.771 46.427 52.386 1.00 11.84 A
ATOM 1228 CD1 LEU A 308 65.916 45.314 51.814 1.00 14.13 A
ATOM 1229 CD2 LEU A 308 66.521 46.596 53.883 1.00 14.79 A
ATOM 1230 C LEU A 308 70.622 46.774 52.531 1.00 10.29 A
ATOM 1231 0 LEU A 308 71.152 46.828 53.642 1.00 12.51 A
ATOM 1232 N ALA A 309 71.240 46.287 51.458 1.00 10.48 A
ATOM 1233 CA ALA A 309 72.595 45.750 51.543 1.00 11.75 A
ATOM 1234 CB ALA A 309 73.062 45.274 50.165 1.00 11.23 A
ATOM 1235 C ALA A 309 73.571 46.782 52.091 1.00 10.51 A
ATOM 1236 0 ALA A 309 74.357 46.482 52.991 1.00 13.69 A
ATOM 1237 N VAL A 310 73.518 47.996 51.552 1.00 10.84 A
ATOM 1238 CA VAL A 310 74.413 49.065 51.987 1.00 11.11 A
ATOM 1239 CB VAL A 310 74.295 50.297 51.062 1.00 11.70 A
ATOM 1240 CG1 VAL A 310 75.127 51.440 51.612 1.00 10.20 A
ATOM 1241 CG2 VAL A 310 74.759 49.933 49.659 1.00 11.55 A
ATOM 1242 C VAL A 310 74.138 49.492 53.423 1.00 11.21 A
ATOM 1243 0 VAL A 310 75.052 49.572 54.246 1.00 12.91 A
ATOM 1244 N GLY A 311 72.872 49.767 53,720 1.00 10.62 A
ATOM 1245 CA GLY A 311 72.502 50.173 55.066 1.00 11.56 A
ATOM 1246 C GLY A 311 72.999 49.224 56.145 1.00 12.62 A
ATOM 1247 0 GLY A 311 73.504 49.665 57.186 1.00 14.88 A
ATOM 1248 N PHE A 312 72.860 47.921 55.916 1.00 12.70 A
ATOM 1249 CA PHE A 312 73.317 46.949 56.905 1.00 13.65 A
ATOM 1250 CB PHE A 312 72.570 45.624 56.728 1.00 15.16 A
ATOM 1251 CG PHE A 312 71.208 45.619 57.365 1.00 18.03 A
ATOM 1252 CD1 PHE A 312 71.081 45.581 58.744 1.00 19.84 A
ATOM 1253 CD2 PHE A 312 70.062 45.688 56.592 1.00 20.32 A
ATOM 1254 CE1 PHE A 312 69.836 45.613 59.344 1.00 22.38 A
ATOM 1255 CE2 PHE A 312 68.812 45.721 57.186 1.00 20.03 A
ATOM 1256 CZ PHE A 312 68.700 45.684 58.563 1.00 20.47 A
ATOM 1257 C PHE A 312 74.826 46.729 56.884 1.00 13.35 A
ATOM 1258 0 PHE A 312 75.441 46.517 57,929 1.00 14.74 A
ATOM 1259 N LYS A 313 75.423 46.789 55.700 1.00 12.71 A
ATOM 1260 CA LYS A 313 76.868 46.613 55.566 1,00 13.38 A
ATOM 1261 CB LYS A 313 77.250 46.689 54,083 1.00 15.99 A

ATOM 1262 CG LYS A 313 78.730 46.724 53.785 1.00 22.93 A
ATOM 1263 CD LYS A 313 78.959 46.889 52.282 1.00 25.29 A
ATOM 1264 CE LYS A 313 78.294 48.160 51.758 1,00 25.27 A
ATOM 1265 NZ LYS A 313 78.389 48.302 50.274 1.00 29.05 A
ATOM 1266 C LYS A 313 77.642 47.672 56.361 1.00 12.46 A
ATOM 1267 0 LYS A 313 78.683 47.381 56.965 1.00 12.84 A
ATOM 1268 N LEU A 314 77.131 48.900 56.374 1.00 12.42 A
ATOM 1269 CA LEU A 314 77.805 49.989 57.070 1.00 13.48 A
ATOM 1270 CB LEU A 314 77.144 51.325 56.724 1.00 13.87 A
ATOM 1271 CG LEU A 314 77.324 51.741 55.259 1.00 14.33 A
ATOM 1272 CD1 LEU A 314 76.597 53.052 55.003 1.00 16.01 A
ATOM 1273 CD2 LEU A 314 78.807 51.878 54.944 1.00 13.72 A
ATOM 1274 C LEU A 314 77.881 49.817 58.580 1.00 12.65 A
ATOM 1275 0 LEU A 314 78.688 50.479 59.231 1.00 13.97 A
ATOM 1276 N LEU A 315 77.055 48.932 59.132 1.00 14.31 A
ATOM 1277 CA LEU A 315 77.080 48.676 60.569 1.00 14.85 A
ATOM 1278 CB LEU A 315 75.934 47.745 60.968 1.00 15.84 A
ATOM 1279 CG LEU A 315 74.518 48.319 60.892 1.00 15.66 A
ATOM 1280 CD1 LEU A 315 73.509 47.194 61.031 1.00 17.27 A
ATOM 1281 CD2 LEU A 315 74.332 49.363 61.981 1.00 15.76 A
ATOM 1282 C LEU A 315 78.406 48.022 60.929 1.00 15.95 A
ATOM 1283 0 LEU A 315 78.816 48.025 62.093 1.00 17.54 A
ATOM 1284 N GLN A 316 79.079 47.474 59.920 1.00 15.51 A
ATOM 1285 CA GLN A 316 80.358 46.796 60.119 1.00 16.62 A
ATOM 1286 CB GLN A 316 80.480 45.629 59.139 1.00 17.82 A
ATOM 1287 CG GLN A 316 79.479 44.516 59.391 1.00 21.80 A
ATOM 1288 CD GLN A 316 79.541 43.438 58.331 1.00 23.44 A
ATOM 1289 0E1 GLN A 316 80.623 43.073 57.870 1.00 27.65 A
ATOM 1290 NE2 GLN A 316 78.381 42.913 57.944 1.00 26.55 A
ATOM 1291 C GLN A 316 81.594 47.680 60.014 1.00 16.29 A
ATOM 1292 0 GLN A 316 82.711 47.213 60.259 1.00 17.60 A
ATOM 1293 N GLU A 317 81.421 48.946 59.646 1.00 15.99 A
ATOM 1294 CA GLU A 317 82.577 49.826 59.575 1.00 15.21 A
ATOM 1295 CB GLU A 317 82.250 51.121 58.820 1.00 16.42 A
ATOM 1296 CG GLU A 317 81.944 50.874 57.336 1.00 19.70 A
ATOM 1297 CD GLU A 317 82.405 51.999 56.411 1.00 21.20 A
ATOM 1298 OE1 GLU A 317 82.750 53.094 56.899 1.00 21.55 A
ATOM 1299 OE2 GLU A 317 82.412 51.784 55.180 1.00 22.14 A
ATOM 1300 C GLU A 317 83.020 50.103 61.011 1.00 15.05 A
ATOM 1301 0 GLU A 317 82.270 49.861 61.961 1.00 15.02 A
ATOM 1302 N GLU A 318 84.242 50.590 61.170 1.00 16.70 A
ATOM 1303 CA GLU A 318 84.783 50.830 62.502 1.00 17.84 A
ATOM 1304 CB GLU A 318 86.208 51.374 62.387 1.00 21.34 A
ATOM 1305 CG GLU A 318 87.077 51.104 63.611 1.00 24.40 A
ATOM 1306 CD GLU A 318 87.429 49.633 63.775 1.00 26.61 A
ATOM 1307 OE1 GLU A 318 86.513 48.813 64.004 1.00 30.14 A
ATOM 1308 OE2 GLU A 318 88.630 49.295 63.671 1.00 30.43 A
ATOM 1309 C GLU A 318 83.937 51.758 63.367 1.00 17.27 A
ATOM 1310 0 GLU A 318 83.556 52.843 62.935 1.00 17.34 A
ATOM 1311 N HIS A 319 83.651 51.301 64.588 1.00 17.50 A
ATOM 1312 CA HIS A 319 82.875 52.048 65.580 1.00 17.25 A
ATOM 1313 CB HIS A 319 83.651 53.298 65.990 1.00 18.02 A
ATOM 1314 CG HIS A 319 85.082 53.025 66.332 1.00 20.07 A
ATOM 1315 CD2 HIS A 319 85.640 52.129 67.181 1.00 20.84 A
ATOM 1316 ND1 HIS A 319 86.131 53.691 65.735 1.00 22.42 A
ATOM 1317 CE1 HIS A 319 87.273 53.215 66.199 1.00 22.47 A
ATOM 1318 NE2 HIS A 319 87.003 52.266 67.076 1.00 21.94 A
ATOM 1319 C HIS A 319 81.503 52.443 65.065 1.00 16.26 A
ATOM 1320 0 HIS A 319 80.958 53.479 65.442 1.00 18.84 A
ATOM 1321 N CYS A 320 80.924 51.586 64.236 1.00 17.29 A
ATOM 1322 CA CYS A 320 79.641 51.885 63.625 1.00 20.46 A
ATOM 1323 CB CYS A 320 79.839 51.855 62.107 1.00 26.05 A
ATOM 1324 SG CYS A 320 78.587 52.657 61.131 1.00 39.75 A
ATOM 1325 C CYS A 320 78.481 50.965 64.015 1.00 16.87 A
ATOM 1326 0 CYS A 320 77.366 51.150 63.535 1.00 17.20 A
ATOM 1327 N ASP A 321 78.710 49.989 64.890 1.00 16.43 A
ATOM 1328 CA ASP A 321 77.622 49.073 65.228 1,00 16.32 A

ATOM 1329 CB ASP A 321 78.175 47.711 65.650 1.00 18.66 A
ATOM 1330 CG ASP A 321 77.087 46.654 65.750 1.00 21.06 A
ATOM 1331 OD1 ASP A 321 75.950 46.924 65.300 1.00 20.72 A
ATOM 1332 OD2 ASP A 321 77.365 45.553 66.265 1.00 23.53 A
ATOM 1333 C ASP A 321 76.644 49.572 66.281 1.00 15.84 A
ATOM 1334 0 ASP A 321 76.808 49.322 67.479 1.00 17.01 A
ATOM 1335 N ILE A 322 75.605 50.263 65.827 1.00 15.83 A
ATOM 1336 CA ILE A 322 74.606 50.790 66.744 1.00 15.80 A
ATOM 1337 CB ILE A 322 73.722 51.855 66.068 1.00 15.39 A
ATOM 1338 CG2 ILE A 322 74.569 53.053 65.691 1.00 13.98 A
ATOM 1339 CG1 ILE A 322 73.023 51.265 64.846 1.00 15.35 A
ATOM 1340 CD1 ILE A 322 72.015 52.209 64.217 1.00 16.08 A
ATOM 1341 C ILE A 322 73.703 49.708 67.323 1.00 15.57 A
ATOM 1342 0 ILE A 322 72.931 49.972 68.245 1.00 17.56 A
ATOM 1343 N PHE A 323 73.797 48.490 66.797 1.00 16.02 A
ATOM 1344 CA PHE A 323 72.962 47.406 67.312 1.00 15.98 A
ATOM 1345 CB PHE A 323 72.281 46.659 66.164 1.00 16.58 A
ATOM 1346 CG PHE A 323 71.388 47.526 65.317 1.00 16.40 A
ATOM 1347 CD1 PHE A 323 70.567 48.476 65.898 1.00 16.29 A
ATOM 1348 CD2 PHE A 323 71.347 47.365 63.940 1.00 16.51 A
ATOM 1349 CE1 PHE A 323 69.720 49.250 65.124 1.00 18.27 A
ATOM 1350 CE2 PHE A 323 70.501 48.137 63.160 1.00 17.82 A
ATOM 1351 CZ PHE A 323 69.688 49.078 63.751 1.00 16.29 A
ATOM 1352 C PHE A 323 73.748 46.423 68.173 1.00 17.19 A
ATOM 1353 0 PHE A 323 73.273 45.331 68.483 1.00 17.72 A
ATOM 1354 N MET A 324 74.947 46.831 68.570 1.00 18.03 A
ATOM 1355 CA MET A 324 75.816 46.000 69.388 1.00 19.94 A
ATOM 1356 CB MET A 324 77.042 46.803 69.827 1.00 21.36 A
ATOM 1357 CG MET A 324 77.957 46.047 70.774 1.00 24.50 A
ATOM 1358 SD MET A 324 79.345 47.044 71.328 1.00 26.18 A
ATOM 1359 CE MET A 324 78.534 48.149 72.480 1.00 27.81 A
ATOM 1360 C MET A 324 75.143 45.410 70.624 1.00 19.21 A
ATOM 1361 0 MET A 324 75.384 44.256 70.970 1.00 21.99 A
ATOM 1362 N ASN A 325 74.298 46.188 71.292 1.00 19.81 A
ATOM 1363 CA ASN A 325 73.662 45.678 72.495 1.00 19.35 A
ATOM 1364 CB ASN A 325 73.616 46.774 73.558 1.00 20.17 A
ATOM 1365 CG ASN A 325 75.001 47.113 74.081 1.00 21.13 A
ATOM 1366 0D1 ASN A 325 75.766 46.220 74.451 1.00 23.54 A
ATOM 1367 ND2 ASN A 325 75.333 48.396 74.110 1.00 24.33 A
ATOM 1368 C ASN A 325 72.301 45.017 72.327 1.00 19.91 A
ATOM 1369 0 ASN A 325 71.643 44.676 73.311 1.00 21.09 A
ATOM 1370 N LEU A 326 71.870 44.837 71.084 1.00 18.05 A
ATOM 1371 CA LEU A 326 70.614 44.144 70.841 1.00 17.42 A
ATOM 1372 CB LEU A 326 70.075 44.455 69.442 1.00 19.27 A
ATOM 1373 CG LEU A 326 69.235 45.724 69.239 1.00 21.05 A
ATOM 1374 CD1 LEU A 326 67.943 45.606 70.015 1.00 24.37 A
ATOM 1375 CD2 LEU A 326 70.017 46.947 69.681 1.00 21.56 A
ATOM 1376 C LEU A 326 70.984 42.666 70.926 1.00 16.65 A
ATOM 1377 0 LEU A 326 72.110 42.290 70.594 1.00 17.37 A
ATOM 1378 N THR A 327 70.060 41.833 71.390 1.00 16.29 A
ATOM 1379 CA THR A 327 70.334 40.405 71.468 1.00 17.21 A
ATOM 1380 CB THR A 327 69.244 39.651 72.261 1.00 17.28 A
ATOM 1381 OG1 THR A 327 67.975 39.818 71.616 1.00 19.02 A
ATOM 1382 CG2 THR A 327 69.157 40.179 73.683 1.00 18.40 A
ATOM 1383 C THR A 327 70.330 39.893 70.037 1.00 17.93 A
ATOM 1384 0 THR A 327 69.813 40.566 69.144 1.00 16.92 A
ATOM 1385 N LYS A 328 70.912 38.719 69.808 1.00 19.42 A
ATOM 1386 CA LYS A 328 70.933 38.152 68.462 1.00 20.02 A
ATOM 1387 CB LYS A 328 71.576 36.762 68.465 1.00 21.62 A
ATOM 1388 CG LYS A 328 73.087 36.776 68.551 1.00 23.65 A
ATOM 1389 CD LYS A 328 73.709 37.382 67.301 1.00 26.20 A
ATOM 1390 CE LYS A 328 75.228 37.386 67.393 1.00 26.95 A
ATOM 1391 NZ LYS A 328 75.879 37.925 66.164 1.00 28.12 A
ATOM 1392 C LYS A 328 69.506 38.052 67.941 1.00 19.58 A
ATOM 1393 0 LYS A 328 69.239 38.330 66.775 1.00 19.40 A
ATOM 1394 N LYS A 329 68.594 37.658 68.824 1.00 19.59 A
ATOM 1395 CA LYS A 329 67.186 37.523 68.479 1.00 19.05 A

ATOM 1396 CB LYS A 329 66.411 36.990 69.683 1.00 19.28 A
ATOM 1397 CG LYS A 329 64.908 36.962 69.507 1.00 20.82 A
ATOM 1398 CD LYS A 329 64.484 35.930 68.485 1.00 22.28 A
ATOM 1399 CE LYS A 329 62.974 35.898 68.358 1.00 23.96 A
ATOM 1400 NZ LYS A 329 62.320 35.634 69.674 1.00 27.59 A
ATOM 1401 C LYS A 329 66.592 38.858 68.030 1.00 18.15 A
ATOM 1402 0 LYS A 329 65.920 38.931 66.999 1.00 19.42 A
ATOM 1403 N GLN A 330 66.828 39.911 68.807 1.00 16.59 A
ATOM 1404 CA GLN A 330 66.309 41.230 68.450 1.00 16.88 A
ATOM 1405 CB GLN A 330 66.648 42.262 69.526 1.00 16.08 A
ATOM 1406 CG GLN A 330 65.909 42.082 70.841 1.00 16.71 A
ATOM 1407 CD GLN A 330 66.338 43.108 71.867 1.00 14.27 A
ATOM 1408 OE1 GLN A 330 67.525 43.260 72.136 1.00 15.49 A
ATOM 1409 NE2 GLN A 330 65.374 43.820 72.445 1.00 17.15 A
ATOM 1410 C GLN A 330 66.897 41.685 67.116 1.00 15.88 A
ATOM 1411 0 GLN A 330 66.201 42.277 66.299 1.00 16.92 A
ATOM 1412 N ARG A 331 68.182 41.414 66.907 1.00 16.85 A
ATOM 1413 CA ARG A 331 68.844 41.796 65.665 1.00 17.39 A
ATOM 1414 CB ARG A 331 70.321 41.396 65.688 1.00 17.37 A
ATOM 1415 CG ARG A 331 71.218 42.258 66.540 1.00 20.72 A
ATOM 1416 CD ARG A 331 72.345 42.808 65.683 1.00 23.49 A
ATOM 1417 NE ARG A 331 73.583 42.991 66.427 1.00 27.97 A
ATOM 1418 CZ ARG A 331 74.657 43.600 65.935 1.00 23.57 A
ATOM 1419 NH1 ARG A 331 74.637 44.088 64.699 1.00 28.18 A
ATOM 1420 NH2 ARG A 331 75.753 43.706 66.669 1.00 27.70 A
ATOM 1421 C ARG A 331 68.180 41.121 64.472 1.00 17.42 A
ATOM 1422 0 ARG A 331 67.972 41.745 63.430 1.00 17.91 A
ATOM 1423 N GLN A 332 67.862 39.838 64.625 1.00 17.79 A
ATOM 1424 CA GLN A 332 67.220 39.079 63.559 1.00 19.81 A
ATOM 1425 CB GLN A 332 67.100 37.605 63.947 1.00 20.35 A
ATOM 1426 CG GLN A 332 68.418 36.866 64.027 1.00 24.70 A
ATOM 1427 CD GLN A 332 68.222 35.378 64.239 1.00 26.39 A
ATOM 1428 OE1 GLN A 332 69.186 34.620 64.341 1.00 29.23 A
ATOM 1429 NE2 GLN A 332 66.963 34.951 64.300 1.00 28.58 A
ATOM 1430 C GLN A 332 65.834 39.628 63.264 1.00 18.60 A
ATOM 1431 0 GLN A 332 65.459 39.805 62.105 1.00 19.38 A
ATOM 1432 N THR A 333 65.076 39.888 64.323 1.00 17.51 A
ATOM 1433 CA THR A 33=3 63.726 40.416 64.187 1.00 17.83 A
ATOM 1434 CB THR A 333 63.046 40.546 65,561 1.00 18.75 A
ATOM 1435 001 THR A 333 63.038 39.267 66.208 1.00 20.67 A
ATOM 1436 CG2 THR A 333 61.615 41.037 65.404 1.00 19.79 A
ATOM 1437 C THR A 333 63.756 41.783 63.520 1.00 15.62 A
ATOM 1438 0 THR A 333 62.942 42.070 62.639 1.00 15.43 A
ATOM 1439 N LEU A 334 64.696 42.625 63.938 1.00 15.88 A
ATOM 1440 CA LEU A 334 64.812 43.953 63.359 1.00 14.77 A
ATOM 1441 CB LEU A 334 65.869 44.779 64.099 1.00 15.78 A
ATOM 1442 CG LEU A 334 66.103 46.180 63.524 1.00 17.62 A
ATOM 1443 CD1 LEU A 334 66.384 47.165 64.643 1.00 18.10 A
ATOM 1444 CD2 LEU A 334 67.250 46.131 62.524 1.00 18.31 A
ATOM 1445 C LEU A 334 65.167 43.861 61.878 1.00 14.30 A
ATOM 1446 0 LEU A 334 64.592 44.574 61.058 1.00 15.17 A
ATOM 1447 N ARG A 335 66.107 42.981 61.535 1.00 14.04 A
ATOM 1448 CA ARG A 335 66.504 42.826 60.135 1.00 14.97 A
ATOM 1449 CB ARG A 335 67.600 41.767 60.001 1.00 17.59 A
ATOM 1450 CG ARG A 335 68.266 41.758 58.632 1.00 21.60 A
ATOM 1451 CD ARG A 335 69.141 40.530 58.433 1.00 25.47 A
ATOM 1452 NE ARG A 335 69.938 40.628 57.211 1.00 27.46 A
ATOM 1453 CZ ARG A 335 71.033 41.374 57.095 1.00 27.50 A
ATOM 1454 NH1 ARG A 335 71.464 42.084 58.129 1.00 28.88 A
ATOM 1455 NH2 ARG A 335 71.698 41.413 55.949' 1.00 28.61 A
ATOM 1456 C ARG A 335 65.299 42.427 59.278 1.00 14.29 A
ATOM 1457 0 ARG A 335 65.075 42.988 58.207 1.00 14.75 A
ATOM 1458 N LYS A 336 64.520 41.462 59.754 1.00 14.67 A
ATOM 1459 CA LYS A 336 63.347 41.012 59.014 1.00 15.65 A
ATOM 1460 CB LYS A 336 62.658 39.865 59.758 1.00 18.14 A
ATOM 1461 CG LYS A 336 61.402 39.339 59.077 1.00 21.81 A
ATOM 1462 CD LYS A 336 60.729 38.258 59.920 1.00 25.00 A

ATOM 1463 CE LYS A 336 60.349 38.790 61.297 1.00 26.92 A
ATOM 1464 NZ LYS A 336 59.713 37.760 62.169 1.00 28.12 A
ATOM 1465 C LYS A 336 62.358 42.159 58.803 1.00 14.53 A
ATOM 1466 0 LYS A 336 61.824 42.334 57.706 1.00 16.71 A
ATOM 1467 N MET A 337 62.120 42.945 59.849 1.00 13.58 A
ATOM 1468 CA MET A 337 61.191 44.068 59.749 1.00 13.55 A
ATOM 1469 CB MET A 337 60.914 44.654 61.135 1.00 15.02 A
ATOM 1470 CG MET A 337 60.264 43.663 62.078 1.00 16.48 A
ATOM 1471 SD MET A 337 59.612 44.466 63.545 1.00 19.72 A
ATOM 1472 CE MET A 337 61.104 44.844 64.429 1.00 16.64 A
ATOM 1473 C MET A 337 61.691 45.165 58.812 1.00 12.92 A
ATOM 1474 0 MET A 337 60.917 45.711 58.016 1.00 14.56 A
ATOM 1475 N VAL A 338 62.975 45.490 58.905 1.00 13.04 A
ATOM 1476 CA VAL A 338 63.551 46.532 58.056 1.00 12.15 A
ATOM 1477 CB VAL A 338 65.013 46.827 58.463 1.00 11.84 A
ATOM 1478 CG1 VAL A 338 65.638 47.829 57.503 1.00 12.54 A
ATOM 1479 CG2 VAL A 338 65.039 47.387 59.876 1.00 12.66 A
ATOM 1480 C VAL A 338 63.489 46.116 56.592 1.00 10.73 A
ATOM 1481 0 VAL A 338 63.187 46.928 55.720 1.00 11.90 A
ATOM 1482 N ILE A 339 63.763 44.846 56.322 1.00 11.30 A
ATOM 1483 CA ILE A 339 63.704 44.358 54.950 1.00 13.40 A
ATOM 1484 CB ILE A 339 64.062 42.861 54.873 1.00 13.14 A
ATOM 1485 CG2 ILE A 339 63.655 42.289 53.514 1.00 14.90 A
ATOM 1486 CG1 ILE A 339 65.559 42.680 55.121 1.00 13.93 A
ATOM 1487 CD1 ILE A 339 65.997 41.236 55.167 1.00 16.14 A
ATOM 1488 C ILE A 339 62.299 44.569 54.386 1.00 13.56 A
ATOM 1489 0 ILE A 339 62.135 45.103 53.288 1.00 15.35 A
ATOM 1490 N ASP A 340 61.280 44.170 55.143 1.00 13.87 A
ATOM 1491 CA ASP A 340 59.908 44.331 54.669 1.00 14.97 A
ATOM 1492 CB ASP A 340 58.922 43.698 55.654 1.00 19.82 A
ATOM 1493 CG ASP A 340 59.130 42.203 55.801 1.00 25.42 A
ATOM 1494 OD1 ASP A 340 59.288 41.520 54.765 1.00 26.97 A
ATOM 1495 OD2 ASP A 340 59.129 41.707 56.950 1.00 28.82 A
ATOM 1496 C ASP A 340 59.539 45.795 54.441 1.00 13.75 A
ATOM 1497 0 ASP A 340 58.844 46.117 53.484 1.00 15.77 A
ATOM 1498 N MET A 341 60.011 46.686 55.307 1.00 12.85 A
ATOM 1499 CA MET A 341 59.680 48.098 55.153 1.00 12.32 A
ATOM 1500 CB MET A 341 59.991 48.865 56.438 1.00 14.73 A
ATOM 1501 CG MET A 341 58.991 48.565 57.543 1.00 15.86 A
ATOM 1502 SD MET A 341 59.246 49.537 59.028 1.00 21.36 A
ATOM 1503 CE MET A 341 60.489 48.572 59.825 1.00 18.20 A
ATOM 1504 C MET A 341 60.369 48.756 53.962 1.00 12.29 A
ATOM 1505 0 MET A 341 59.743 49.517 53.227 1.00 12.72 A
ATOM 1506 N VAL A 342 61.648 48.463 53.759 1.00 12.16 A
ATOM 1507 CA VAL A 342 62.351 49.061 52.634 1.00 11.71 A
ATOM 1508 CB VAL A 342 63.875 48.852 52.751 1.00 11.04 A
ATOM 1509 CG1 VAL A 342 64.575 49.416 51.518 1.00 11.66 A
ATOM 1510 CG2 VAL A 342 64.392 49.555 53.992 1.00 10.71 A
ATOM 1511 C VAL A 342 61.844 48.497 51.306 1.00 11.79 A
ATOM 1512 0 VAL A 342 61.702 49.232 50.325 1.00 12.78 A
ATOM 1513 N LEU A 343 61.555 47.202 51.262 1.00 12.33 A
ATOM 1514 CA LEU A 343 61.051 46.620 50.024 1.00 13.51 A
ATOM 1515 CB LEU A 343 60.976 45.092 50.129 1.00 14.60 A
ATOM 1516 CG LEU A 343 62.313 44.349 50.121 1.00 17.95 A
ATOM 1517 CD1 LEU A 343 62.067 42.851 50.148 1.00 16.78 A
ATOM 1518 CD2 LEU A 343 63.099 44.733 48.880 1.00 19.17 A
ATOM 1519 C LEU A 343 59.673 47.189 49.695 1.00 12.86 A
ATOM 1520 0 LEU A 343 59.256 47.200 48.537 1.00 14.40 A
ATOM 1521 N ALA A 344 58.967 47.671 50.713 1.00 13.89 A
ATOM 1522 CA ALA A 344 57.639 48.239 50.503 1.00 14.80 A
ATOM 1523 CB ALA A 344 56.879 48.312 51.829 1.00 16.07 A
ATOM 1524 C ALA A 344 57.710 49.622 49.859 1.00 14.06 A
ATOM 1525 0 ALA A 344 56.681 50.194 49.489 1.00 16.21 A
ATOM 1526 N THR A 345 58.917 50.166 49.718 1.00 13.00 A
ATOM 1527 CA THR A 345 59.055 51.475 49.091 1.00 13.04 A
ATOM 1528 CE THR A 345 60.222 52.305 49.695 1.00 11.75 A
ATOM 1529 OG1 THR A 345 61.466 51.626 49.496 1.00 14.32 A

ATOM 1530 CG2 THR A 345 59.995 52.529 51.191 1.00 12.17 A
ATOM 1531 C THR A 345 59.238 51.344 47.578 1.00 13.13 A
ATOM 1532 0 THR A 345 59.377 52.343 46.877 1.00 15.04 A
ATOM 1533 N ASP A 346 59.235 50.111 47.080 1.00 15.13 A
ATOM 1534 CA ASP A 346 59.343 49.871 45.641 1.00 16.12 A
ATOM 1535 CB ASP A 346 59.647 48.394 45.361 1.00 17.10 A
ATOM 1536 CG ASP A 346 59.748 48.080 43.872 1.00 19.43 A
ATOM 1537 OD1 ASP A 346 59.300 48.898 43.046 1.00 20.70 A
ATOM 1538 OD2 ASP A 346 60.271 47.002 43.525 1.00 18.76 A
ATOM 1539 C ASP A 346 57.969 50.225 45.071 1.00 16.26 A
ATOM 1540 0 ASP A 346 56.984 49.537 45.342 1.00 16.33 A
ATOM 1541 N MET A 347 57.903 51.298 44.290 1.00 18.28 A
ATOM 1542 CA MET A 347 56.640 51.741 43.705 1.00 21.29 A
ATOM 1543 CB MET A 347 56.858 52.997 42.861 1.00 25.96 A
ATOM 1544 CG MET A 347 56.796 54.281 43.657 1.00 31.87 A
ATOM 1545 SD MET A 347 55.132 54.562 44.281 1.00 34.25 A
ATOM 1546 CE MET A 347 54.435 55.496 42.943 1.00 34.55 A
ATOM 1547 C MET A 347 55.913 50.700 42.864 1.00 21.27 A
ATOM 1548 0 MET A 347 54.694 50.768 42.719 1.00 21.62 A
ATOM 1549 N SER A 348 56.648 49.740 42.311 1.00 21.21 A
ATOM 1550 CA SER A 348 56.023 48.712 41.486 1.00 22.59 A
ATOM 1551 CB SER A 348 57.092 47.866 40.787 1.00 22.39 A
ATOM 1552 OG SER A 348 57.782 47.038 41.705 1.00 26.83 A
ATOM 1553 C SER A 348 55.105 47.810 42.313 1.00 23.21 A
ATOM 1554 0 SER A 348 54.302 47.051 41.762 1.00 24.23 A
ATOM 1555 N LYS A 349 55.214 47.905 43.635 1.00 22.03 A
ATOM 1556 CA LYS A 349 54.401 47.091 44.534 1.00 21.97 A
ATOM 1557 CB LYS A 349 55.267 46.529 45.665 1.00 22.74 A
ATOM 1558 CG LYS A 349 56.581 45.901 45.223 1.00 23.77 A
ATOM 1559 CD LYS A 349 56.363 44.711 44.308 1.00 24.27 A
ATOM 1560 CE LYS A 349 57.693 44.118 43.855 1.00 24.59 A
ATOM 1561 NZ LYS A 349 57.493 42.948 42.957 1.00 25.73 A
ATOM 1562 C LYS A 349 53.262 47.897 45.157 1.00 20.67 A
ATOM 1563 0 LYS A 349 52.535 47.394 46.009 1.00 21.69 A
ATOM 1564 N HIS A 350 53.107 49.145 44.730 1.00 19.53 A
ATOM 1565 CA HIS A 350 52.074 50.008 45.287 1.00 19.27 A
ATOM 1566 CB HIS A 350 52.067 51.360 44.579 1.00 19.99 A
ATOM 1567 CG HIS A 350 50.965 52.263 45.036 1.00 22.05 A
ATOM 1568 CD2 HIS A 350 49.843 52.689 44.408 1.00 22.13 A
ATOM 1569 ND1 HIS A 350 50.915 52.791 46.308 1.00 24.88 A
ATOM 1570 CE1 HIS A 350 49.810 53.504 46.445 1.00 24.84 A
ATOM 1571 NE2 HIS A 350 49.142 53.458 45.307 1.00 25.02 A
ATOM 1572 C HIS A 350 50.650 49.455 45.288 1.00 19.80 A
ATOM 1573 0 HIS A 350 49.968 49.511 46.308 1.00 20.38 A
ATOM 1574 N MET A 351 50.198 48.933 44.154 1.00 20.02 A
ATOM 1575 CA MET A 351 48.838 48.412 44.063 1.00 21.53 A
ATOM 1576 CB MET A 351 48.521 48.004 42.621 1.00 24.75 A
ATOM 1577 CG MET A 351 48.551 49.148 41.617 1.00 28.47 A
ATOM 1578 SD MET A 351 47.436 50.522 42.021 1.00 31.24 A
ATOM 1579 CE MET A 351 48.411 51.925 41.465 1.00 33.64 A
ATOM 1580 C MET A 351 48.576 47.237 44.999 1.00 21.50 A
ATOM 1581 0 MET A 351 47.512 47.159 45.619 1.00 22.29 A
ATOM 1582 N SER A 352 49.536 46.324 45.105 1.00 22.13 A
ATOM 1583 CA SER A 352 49.373 45.163 45.977 1.00 23.14 A
ATOM 1584 CB SER A 352 50.514 44.159 45.764 1.00 23.45 A
ATOM 1585 OG SER A 352 51.763 44.714 46.134 1.00 27.89 A
ATOM 1586 C SER A 352 49.342 45.608 47.434 1.00 23.14 A
ATOM 1587 0 SER A 352 48.574 45.080 48.240 1.00 25.17 A
ATOM 1588 N LEU A 353 50.183 46.583 47.768 1.00 21.80 A
ATOM 1589 CA LEU A 353 50.243 47.110 49.125 1.00 20.98 A
ATOM 1590 CB LEU A 353 51.374 48.126 49.258 1.00 21.58 A
ATOM 1591 CG LEU A 353 52.727 47.642 49.770 1.00 24.48 A
ATOM 1592 CD1 LEU A 353 53.639 48.848 49.925 1.00 23.12 A
ATOM 1593 CD2 LEU A 353 52.559 46.925 51.107 1.00 23.65 A
ATOM 1594 C LEU A 353 48.942 47.785 49.533 1.00 20.87 A
ATOM 1595 0 LEU A 353 48.447 47.573 50.642 1.00 22.05 A
ATOM 1596 N LEU A 354 48.401 48.605 48.636 1.00 20.35 A

ATOM 1597 CA LEU A 354 47.160 49.328 48.899 1.00 19.81 A
ATOM 1598 CB LEU A 354 46.850 50.285 47.748 1.00 19.95 A
ATOM 1599 CG LEU A 354 45.583 51.122 47.927 1.00 19.89 A
ATOM 1600 CD1 LEU A 354 45.732 51.995 49.160 1.00 21.73 A
ATOM 1601 CD2 LEU A 354 45.344 51.966 46.687 1.00 22.26 A
ATOM 1602 C LEU A 354 45.996 48.369 49.074 1.00 20.65 A
ATOM 1603 0 LEU A 354 45.183 48.526 49.985 1.00 21.97 A
ATOM 1604 N ALA A 355 45.920 47.379 48.191 1.00 22.24 A
ATOM 1605 CA ALA A 355 44.848 46.391 48.246 1.00 22.74 A
ATOM 1606 CB ALA A 355 45.022 45.370 47.129 1.00 23.99 A
ATOM 1607 C ALA A 355 44.844 45.693 49.597 1.00 23.62 A
ATOM 1608 0 ALA A 355 43.791 45.512 50.212 1.00 24.76 A
ATOM 1609 N ASP A 356 46.024 45.304 50.065 1.00 25.25 A
ATOM 1610 CA ASP A 356 46.129 44.623 51.345 1.00 25.56 A
ATOM 1611 CB ASP A 356 47.510 43.977 51.489 1.00 27.12 A
ATOM 1612 CG ASP A 356 47.688 42.775 50.572 1.00 28.99 A
ATOM 1613 OD1 ASP A 356 46.830 41.864 50.609 1.00 31.04 A
ATOM 1614 OD2 ASP A 356 48.682 42.735 49.817 1.00 32.31 A
ATOM 1615 C ASP A 356 45.840 45.554 52.520 1.00 25.60 A
ATOM 1616 0 ASP A 356 45.326 45.116 53.549 1.00 27.10 A
ATOM 1617 N LEU A 357 46.157 46.837 52.369 1.00 23.87 A
ATOM 1618 CA LEU A 357 45.892 47.798 53.437 1.00 23.13 A
ATOM 1619 CB LEU A 357 46.545 49.150 53.130 1.00 21.87 A
ATOM 1620 CG LEU A 357 46.349 50.231 54.199 1.00 21.26 A
ATOM 1621 CD1 LEU A 357 46.965 49.768 55.510 1.00 22.31 A
ATOM 1622 CD2 LEU A 357 46.980 51.539 53.747 1.00 22.11 A
ATOM 1623 C LEU A 357 44.381 47.980 53.566 1.00 22.69 A
ATOM 1624 0 LEU A 357 43.856 48.161 54.662 1.00 23.96 A
ATOM 1625 N LYS A 358 43.687 47.933 52.436 1.00 22.86 A
ATOM 1626 CA LYS A 358 42.240 48.082 52.430 1.00 23.64 A
ATOM 1627 CB LYS A 358 41.726 48.186 50.994 1.00 21.78 A
ATOM 1628 CG LYS A 358 41.931 49.542 50.354 1.00 22.04 A
ATOM 1629 CD LYS A 358 41.424 49.537 48.922 1.00 23.04 A
ATOM 1630 CE LYS A 358 41.465 50.927 48.302 1.00 24.68 A
ATOM 1631 NZ LYS A 358 40.536 51.871 48.981 1.00 27.46 A
ATOM 1632 C LYS A 358 41.564 46.912 53.133 1.00 24.81 A
ATOM 1633 0 LYS A 358 40.571 47.095 53.838 1.00 26.10 A
ATOM 1634 N THR A 359 42.101 45.711 52.944 1.00 25.35 A
ATOM 1635 CA THR A 359 41.530 44.527 53.579 1.00 26.13 A
ATOM 1636 CB THR A 359 42.245 43.237 53.135 1.00 25.56 A
ATOM 1637 OG1 THR A 359 42.075 43.053 51.723 1.00 27.97 A
ATOM 1638 CG2 THR A 359 41.665 42.033 53.870 1.00 25.58 A
ATOM 1639 C THR A 359 41.664 44.660 55.089 1.00 26.66 A
ATOM 1640 0 THR A 359 40.722 44.382 55.834 1.00 27.49 A
ATOM 1641 N MET A 360 42.843 45.086 55.531 1.00 26.06 A
ATOM 1642 CA MET A 360 43.116 45.281 56.948 1.00 26.88 A
ATOM 1643 CB MET A 360 44.524 45.852 57.131 1.00 29.78 A
ATOM 1644 CG MET A 360 44.905 46.149 58.571 1.00 32.95 A
ATOM 1645 SD MET A 360 45.119 44.644 59.542 1.00 35.82 A
ATOM 1646 CE MET A 360 46.859 44.343 59.301 1.00 34.82 A
ATOM 1647 C MET A 360 42.091 46.256 57.515 1.00 26.89 A
ATOM 1648 0 MET A 360 41.520 46.033 58.584 1.00 27.79 A
ATOM 1649 N VAL A 361 41.861 47.337 56.777 1.00 25.45 A
ATOM 1650 CA VAL A 361 40.914 48.372 57.172 1.00 25.13 A
ATOM 1651 CB VAL A 361 40.822 49.462 56.099 1.00 24.50 A
ATOM 1652 CG1 VAL A 361 39.820 50.527 56.518 1.00 24.68 A
ATOM 1653 CG2 VAL A 361 42.194 50.049 55.858 1.00 26.07 A
ATOM 1654 C VAL A 361 39.513 47.831 57.404 1.00 24.97 A
ATOM 1655 0 VAL A 361 38.875 48.147 58.406 1.00 25.84 A
ATOM 1656 N GLU A 362 39.031 47.021 56.468 1.00 25.15 A
ATOM 1657 CA GLU A 362 37.694 46.456 56.579 1.00 25.35 A
ATOM 1658 CB GLU A 362 37.391 45.588 55.358 1.00 24.73 A
ATOM 1659 CG GLU A 362 37.456 46.347 54.045 1.00 26.70 A
ATOM 1660 CD GLU A 362 36.929 45.539 52.877 1.00 28.52 A
ATOM 1661 OE1 GLU A 362 37.434 44.419 52.647 1.00 31.92 A
ATOM 1662 OE2 GLU A 362 36.009 46.029 52.187 1.00 31.50 A
ATOM 1663 C GLU A 362 37.498 45.646 57.857 1.00 26.32 A

ATOM 1664 0 GLU A 362 36.404 45.631 58.422 1.00 27.35 A
ATOM 1665 N THR A 363 38.551 44.973 58.308 1.00 26.81 A
ATOM 1666 CA THR A 363 38.477 44.173 59.527 1.00 27.78 A
ATOM 1667 CB THR A 363 39.234 42.836 59.374 1.00 27.80 A
ATOM 1668 OG1 THR A 363 40.613 43.097 59.083 1.00 30.57 A
ATOM 1669 CG2 THR A 363 38.628 42.003 58.254 1.00 27.64 A
ATOM 1670 C THR A 363 39.076 44.936 60.704 1.00 27.86 A
ATOM 1671 0 THR A 363 39.502 44.337 61.693 1.00 28.85 A
ATOM 1672 N LYS A 364 39.102 46.260 60.586 1.00 26.86 A
ATOM 1673 CA LYS A 364 39.643 47.133 61.623 1.00 26.45 A
ATOM 1674 CB LYS A 364 39.460 48.598 61.215 1.00 24.94 A
ATOM 1675 CG LYS A 364 39.867 49.609 62.276 1.00 25.85 A
ATOM 1676 CD LYS A 364 39.610 51.036 61.812 1.00 25.30 A
ATOM 1677 CE LYS A 364 40.031 52.047 62.871 1.00 26.07 A
ATOM 1678 NZ LYS A 364 39.796 53.458 62.446 1.00 27.15 A
ATOM 1679 C LYS A 364 38.983 46.899 62.980 1.00 27.09 A
ATOM 1680 0 LYS A 364 37.763 46.752 63.075 1.00 27.70 A
ATOM 1681 N LYS A 365 39.803 46.867 64.026 1.00 26.80 A
ATOM 1682 CA LYS A 365 39.318 46.671 65.387 1.00 27.07 A
ATOM 1683 CB LYS A 365 39.648 45.260 65.878 1.00 26.82 A
ATOM 1684 CG LYS A 365 39.000 44.149 65.068 1.00 27.96 A
ATOM 1685 CD LYS A 365 39.389 42.780 65.608 1.00 28.82 A
ATOM 1686 CE LYS A 365 38.814 41.661 64.755 1.00 28.89 A
ATOM 1687 NZ LYS A 365 39.274 40.320 65.220 1.00 29.93 A
ATOM 1688 C LYS A 365 40.002 47.694 66.284 1.00 27.63 A
ATOM 1689 0 LYS A 365 41.222 47.864 66.218 1.00 27.67 A
ATOM 1690 N VAL A 366 39.220 48.378 67.113 1.00 26.47 A
ATOM 1691 CA VAL A 366 39.776 49.380 68.011 1.00 25.96 A
ATOM 1692 CB VAL A 366 39.481 50.811 67.508 1.00 24.10 A
ATOM 1693 CG1 VAL A 366 40.035 50.994 66.103 1.00 24.03 A
ATOM 1694 CG2 VAL A 366 37.984 51.075 67.530 1.00 24.00 A
ATOM 1695 C VAL A 366 39.246 49.256 69.434 1.00 26.37 A
ATOM 1696 0 VAL A 366 38.172 48.694 69.664 1.00 28.22 A
ATOM 1697 N THR A 367 40.017 49.781 70.383 1.00 26.43 A
ATOM 1698 CA THR A 367 39.653 49.771 71.798 1.00 27.54 A
ATOM 1699 CB THR A 367 40.452 48.719 72.598 1.00 27.71 A
ATOM 1700 OGl THR A 367 41.857 48.908 72.377 1.00 28.47 A
ATOM 1701 CG2 THR A 367 40.049 47.314 72.186 1.00 28.59 A
ATOM 1702 C THR A 367 39.955 51.138 72.397 1.00 26.70 A
ATOM 1703 0 THR A 367 41.101 51.589 72.377 1.00 26.71 A
ATOM 1704 N GLY A 370 40.510 55.504 71.435 1.00 25.74 A
ATOM 1705 CA GLY A 370 40.309 55.041 70.077 1.00 27.88 A
ATOM 1706 C GLY A 370 41.547 54.357 69.535 1.00 28.02 A
ATOM 1707 0 GLY A 370 41.620 54.032 68.351 1.00 29.88 A
ATOM 1708 N VAL A 371 41.968 53.301 69.825 1.00 27.21 A
ATOM 1709 CA VAL A 371 43.286 52.750 69.527 1.00 25.97 A
ATOM 1710 CB VAL A 371 44.092 52.494 70.818 1.00 24.86 A
ATOM 1711 CG1 VAL A 371 45.403 51.799 70.481 1.00 23.86 A
ATOM 1712 CG2 VAL A 371 44.360 53.808 71.534 1.00 25.07 A
ATOM 1713 C VAL A 371 43.163 51.438 68.764 1.00 25.29 A
ATOM 1714 0 VAL A 371 42.406 50.548 69.153 1.00 25.40 A
ATOM 1715 N LEU A 372 43.917 51.321 67.677 1.00 24.10 A
ATOM 1716 CA LEU A 372 43.888 50.120 66.856 1.00 24.56 A
ATOM 1717 CB LEU A 372 44.783 50.301 65.627 1.00 24.29 A
ATOM 1718 CG LEU A 372 44.276 51.289 64.579 1.00 24.78 A
ATOM 1719 CD1 LEU A 372 45.336 51.496 63.512 1.00 25.56 A
ATOM 1720 CD2 LEU A 372 42.996 50.758 63.965 1.00 24.78 A
ATOM 1721 C LEU A 372 44.326 48.877 67.614 1.00 24.79 A
ATOM 1722 0 LEU A 372 45.168 48.942 68.512 1.00 25.30 A
ATOM 1723 N LEU A 373 43.741 47.744 67.243 1.00 24.44 A
ATOM 1724 CA LEU A 373 44.075 46.468 67.853 1.00 25.16 A
ATOM 1725 CB LEU A 373 42.808 45.717 68.271 1.00 25.33 A
ATOM 1726 CG LEU A 373 42.048 46.268 69.475 1.00 24.87 A
ATOM 1727 CD1 LEU A 373 40.831 45.395 69.737 1.00 26.12 A
ATOM 1728 CD2 LEU A 373 42.962 46.292 70.695 1.00 26.02 A
ATOM 1729 C LEU A 373 44.856 45.628 66.853 1.00 25.04 A
ATOM 1730 0 LEU A 373 44.280 45.046 65.931 1.00 25.70 A

ATOM 1731 N LEU A 374 46.171 45.590 67.039 1.00 24.97 A
ATOM 1732 CA LEU A 374 47.066 44.823 66.183 1.00 25.46 A
ATOM 1733 CB LEU A 374 48.139 45.736 65.594 1.00 23.98 A
ATOM 1734 CG LEU A 374 47.590 46.881 64.741 1.00 23.68 A
ATOM 1735 CD1 LEU A 374 48.740 47.699 64.173 1.00 24.31 A
ATOM 1736 CD2 LEU A 374 46.739 46.314 63.620 1.00 22.68 A
ATOM 1737 C LEU A 374 47.700 43.750 67.059 1.00 26.45 A
ATOM 1738 0 LEU A 374 48.868 43.837 67.444 1.00 27.79 A
ATOM 1739 N ASP A 375 46.897 42.738 67.363 1.00 26.61 A
ATOM 1740 CA ASP A 375 47.284 41.621 68.213 1.00 27.28 A
ATOM 1741 CB ASP A 375 46.082 40.688 68.379 1.00 27.38 A
ATOM 1742 CG ASP A 375 45.435 40.336 67.052 1.00 28.74 A
ATOM 1743 OD1 ASP A 375 46.094 39.674 66.223 1.00 31.27 A
ATOM 1744 OD2 ASP A 375 44.269 40.731 66.832 1.00 31.82 A
ATOM 1745 C ASP A 375 48.499 40.803 67.789 1.00 27.98 A
ATOM 1746 0 ASP A 375 49.298 40.403 68.637 1.00 28.80 A
ATOM 1747 N ASN A 376 48.641 40.542 66.493 1.00 26.01 A
ATOM 1748 CA ASN A 376 49.764 39.737 66.027 1.00 24.81 A
ATOM 1749 CB ASN A 376 49.255 38.518 65.251 1.00 25.46 A
ATOM 1750 CG ASN A 376 48.438 38.898 64.030 1.00 26.39 A
ATOM 1751 0D1 ASN A 376 48.913 39.613 63.149 1.00 30.51 A
ATOM 1752 ND2 ASN A 376 47.201 38.414 63.971 1.00 28.24 A
ATOM 1753 C ASN A 376 50.785 40.483 65.179 1.00 25.28 A
ATOM 1754 0 ASN A 376 50.506 41.552 64.633 1.00 25.65 A
ATOM 1755 N TYR A 377 51.972 39.892 65.081 1.00 23.91 A
ATOM 1756 CA TYR A 377 53.079 40.451 64.312 1.00 23.31 A
ATOM 1757 CB TYR A 377 54.264 39.486 64.344 1.00 22.27 A
ATOM 1758 CG TYR A 377 55.371 39.853 63.385 1.00 21.76 A
ATOM 1759 CD1 TYR A 377 56.234 40.904 63.661 1.00 22.68 A
ATOM 1760 CE1 TYR A 377 57.222 41.271 62.764 1.00 22.82 A
ATOM 1761 CD2 TYR A 377 55.527 39.173 62.183 1.00 20.93 A
ATOM 1762 CE2 TYR A 377 56.510 39.534 61.280 1.00 22.74 A
ATOM 1763 CZ TYR A 377 57.354 40.585 61.575 1.00 22.76 A
ATOM 1764 OH TYR A 377 58.319 40.967 60.669 1.00 24.97 A
ATOM 1765 C TYR A 377 52.692 40.718 62.860 1.00 22.84 A
ATOM 1766 0 TYR A 377 52.975 41.788 62.315 1.00 23.96 A
ATOM 1767 N THR A 378 52.051 39.736 62.237 1.00 23.38 A
ATOM 1768 CA THR A 378 51.636 39.846 60.845 1.00 23.80 A
ATOM 1769 CB THR A 378 50.734 38.661 60.446 1.00 25.48 A
ATOM 1770 OG1 THR A 378 51.446 37.433 60.651 1.00 29.10 A
ATOM 1771 CG2 THR A 378 50.325 38.772 58.980 1.00 26.98 A
ATOM 1772 C THR A 378 50.897 41.145 60.543 1.00 22.83 A
ATOM 1773 0 THR A 378 51.166 41.798 59.533 1.00 24.33 A
ATOM 1774 N ASP A 379 49.963 41.516 61.412 1.00 22.53 A
ATOM 1775 CA ASP A 379 49.197 42.742 61.220 1.00 22.43 A
ATOM 1776 CB ASP A 379 47.948 42.736 62.108 1.00 24.36 A
ATOM 1777 CG ASP A 379 46.893 41.756 61.626 1.00 27.03 A
ATOM 1778 0D1 ASP A 379 47.198 40.550 61.518 1.00 30.79 A
ATOM 1779 OD2 ASP A 379 45.756 42.194 61.355 1.00 30.32 A
ATOM 1780 C ASP A 379 50.028 43.985 61.519 1.00 21.23 A
ATOM 1781 0 ASP A 379 49.892 45.006 60.845 1.00 21.56 A
ATOM 1782 N ARG A 380 50.889 43.902 62.527 1.00 20.84 A
ATOM 1783 CA ARG A 380 51.716 45.042 62.885 1.00 19.89 A
ATOM 1784 CB ARG A 380 52.470 44.779 64.195 1.00 20.64 A
ATOM 1785 CG ARG A 380 51.550 44.664 65.414 1.00 22.66 A
ATOM 1786 CD ARG A 380 52.262 45.041 66.703 1.00 23.15 A
ATOM 1787 NE ARG A 380 53.374 44.152 67.013 1.00 25.09 A
ATOM 1788 CZ ARG A 380 53.245 42.870 67.342 1.00 24.87 A
ATOM 1789 NH1 ARG A 380 52.041 42.316 67.408 1.00 25.24 A
ATOM 1790 NH2 ARG A 380 54.322 42.143 67.608 1.00 27.04 A
ATOM 1791 C ARG A 380 52.697 45.390 61.773 1.00 18.86 A
ATOM 1792 0 ARG A 380 52.841 46.557 61.418 1.00 19.07 A
ATOM 1793 N ILE A 381 53.361 44.386 61.212 1.00 18.19 A
ATOM 1794 CA ILE A 381 54.318 44.647 60.140 1.00 19.09 A
ATOM 1795 CB ILE A 381 55.142 43.382 59.787 1.00 18.39 A
ATOM 1796 CG2 ILE A 381 54.259 42.347 59.108 1.00 20.45 A
ATOM 1797 CG1 ILE A 381 56.316 43.769 58.885 1.00 19.91 A

ATOM 1798 CD1 ILE A 381 57.230 44.806 59.501 1.00 21.55 A
ATOM 1799 C ILE A 381 53.608 45.166 58.889 1.00 18.80 A
ATOM 1800 0 ILE A 381 54.187 45.918 58.106 1.00 19.63 A
ATOM 1801 N GLN A 382 52.353 44.773 58.705 1.00 19.73 A
ATOM 1802 CA GLN A 382 51.581 45.232 57.553 1.00 20.03 A
ATOM 1803 CB GLN A 382 50.231 44.515 57.498 1.00 21.35 A
ATOM 1804 CG GLN A 382 49.439 44.788 56.232 1.00 24.59 A
ATOM 1805 CD GLN A 382 50.016 44.085 55.021 1.00 27.65 A
ATOM 1806 OE1 GLN A 382 49.460 44.157 53.926 1.00 31.64 A
ATOM 1807 NE2 GLN A 382 51.136 43.396 55.211 1.00 29.16 A
ATOM 1808 C GLN A 382 51.354 46.735 57.692 1.00 20.05 A
ATOM 1809 0 GLN A 382 51.412 47.480 56.719 1.00 21.36 A
ATOM 1810 N VAL A 383 51.092 47.177 58.916 1.00 19.07 A
ATOM 1811 CA VAL A 383 50.872 48.591 59.168 1.00 18.28 A
ATOM 1812 CB VAL A 383 50.283 48.819 60.579 1.00 18.81 A
ATOM 1813 CG1 VAL A 383 50.235 50.306 60.891 1.00 18.02 A
ATOM 1814 CG2 VAL A 383 48.879 48.225 60.645 1.00 20.64 A
ATOM 1815 C VAL A 383 52.185 49.361 59.019 1.00 17.24 A
ATOM 1816 0 VAL A 383 52.222 50.426 58.406 1.00 17.10 A
ATOM 1817 N LEU A 384 53.265 48.808 59.559 1.00 16.44 A
ATOM 1818 CA LEU A 384 54.569 49.457 59.471 1.00 16.62 A
ATOM 1819 CB LEU A 384 55.603 48.692 60.301 1.00 18.15 A
ATOM 1820 CG LEU A 384 55.409 48.726 61.818 1.00 20.02 A
ATOM 1821 CD1 LEU A 384 56.434 47.821 62.472 1.00 22.12 A
ATOM 1822 CD2 LEU A 384 55.550 50.155 62.332 1.00 21.97 A
ATOM 1823 C LEU A 384 55.051 49.580 58.032 1.00 15.97 A
ATOM 1824 0 LEU A 384 55.636 50.595 57.659 1.00 18.19 A
ATOM 1825 N ARG A 385 54.816 48.555 57.218 1.00 15.80 A
ATOM 1826 CA ARG A 385 55.245 48.620 55.824 1.00 16.02 A
ATOM 1827 CB ARG A 385 55.020 47.278 55.117 1.00 18.31 A
ATOM 1828 CG ARG A 385 53.577 46.903 54.890 1.00 24.03 A
ATOM 1829 CD ARG A 385 53.460 45.456 54.414 1.00 26.25 A
ATOM 1830 NE ARG A 385 54.272 45.201 53.229 1.00 27.51 A
ATOM 1831 CZ ARG A 385 54.291 44.047 52.569 1.00 27.90 A
ATOM 1832 NH1 ARG A 385 53.542 43.032 52.980 1.00 30.71 A
ATOM 1833 NH2 ARG A 385 55.051 43.911 51.490 1.00 28.89 A
ATOM 1834 C ARG A 385 54.482 49.733 55.110 1.00 15.36 A
ATOM 1835 0 ARG A 385 55.051 50.477 54.314 1.00 16.46 A
ATOM 1836 N ASN A 386 53.195 49.864 55.405 1.00 16.28 A
ATOM 1837 CA ASN A 386 52.400 50.910 54.777 1.00 16.79 A
ATOM 1838 CB ASN A 386 50.910 50.605 54.941 1.00 18.73 A
ATOM 1839 CG ASN A 386 50.412 49.613 53.905 1.00 22.47 A
ATOM 1840 OD1 ASN A 386 50.233 49.960 52.737 1.00 24.09 A
ATOM 1841 ND2 ASN A 386 50.207 48.369 54.321 1.00 24.87 A
ATOM 1842 C ASN A 386 52.737 52.293 55.327 1.00 15.78 A
ATOM 1843 0 ASN A 386 52.603 53.298 54.627 1.00 15.65 A
ATOM 1844 N MET A 387 53.190 52.344 56.576 1.00 16.58 A
ATOM 1845 CA MET A 387 53.551 53.613 57.191 1.00 15.91 A
ATOM 1846 CB MET A 387 53.853 53.425 58.675 1.00 17.75 A
ATOM 1847 CG MET A 387 54.162 54.722 59.398 1.00 18.40 A
ATOM 1848 SD MET A 387 54.685 54.438 61.097 1.00 21.99 A
ATOM 1849 CE MET A 387 53.127 54.046 61.856 1.00 19.64 A
ATOM 1850 C MET A 387 54.765 54.212 56.502 1.00 15.41 A
ATOM 1851 0 MET A 387 54.766 55.390 56.146 1.00 15.59 A
ATOM 1852 N VAL A 388 55.804 53.402 56.315 1.00 16.37 A
ATOM 1853 CA VAL A 388 57.016 53.888 55.663 1.00 15.81 A
ATOM 1854 CB VAL A 388 58.155 52.853 55.749 1.00 17.43 A
ATOM 1855 CG1 VAL A 388 59.375 53.356 54.990 1.00 18.43 A
ATOM 1856 CG2 VAL A 388 58.521 52.618 57.208 1.00 18.17 A
ATOM 1857 C VAL A 388 56.719 54.211 54.203 1.00 15.38 A
ATOM 1858 0 VAL A 388 57.279 55.153 53.641 1.00 14.00 A
ATOM 1859 N HIS A 389 55.828 53.433 53.594 1.00 14.57 A
ATOM 1860 CA HIS A 389 55.440 53.670 52.207 1.00 15.93 A
ATOM 1861 CB HIS A 389 54.499 52.555 51.733 1.00 17.19 A
ATOM 1862 CG HIS A 389 53.998 52.730 50.332 1.00 18.34 A
ATOM 1863 CD2 HIS A 389 52.760 52.575 49.805 1.00 19.82 A
ATOM 1864 ND1 HIS A 389 54.818 53.086 49.284 1.00 19.86 A

ATOM 1865 CE1 HIS A 389 54.107 53.144 48.171 1.00 21.07 A
ATOM 1866 NE2 HIS A 389 52.855 52.838 48.461 1.00 19.96 A
ATOM 1867 C HIS A 389 54.758 55.040 52.124 1.00 14.29 A
ATOM 1868 0 HIS A 389 55.019 55.819 51.206 1.00 15.16 A
ATOM 1869 N CYS A 390 53.896 55.334 53.096 1.00 14.65 A
ATOM 1870 CA CYS A 390 53.203 56.624 53.146 1.00 14.68 A
ATOM 1871 CB CYS A 390 52.181 56.641 54.284 1.00 15.22 A
ATOM 1872 SG CYS A 390 50.637 55.757 53.923 1.00 23.86 A
ATOM 1873 C CYS A 390 54.195 57.764 53.358 1.00 12.43 A
ATOM 1874 0 CYS A 390 54.095 58.819 52.733 1.00 12.71 A
ATOM 1875 N ALA A 391 55.149 57.550 54.257 1.00 12.70 A
ATOM 1876 CA ALA A 391 56.159 58.558 54.539 1.00 11.72 A
ATOM 1877 CB ALA A 391 57.104 58.053 55.621 1.00 10.94 A
ATOM 1878 C ALA A 391 56.941 58.873 53.258 1.00 11.76 A
ATOM 1879 0 ALA A 391 57.231 60.032 52.967 1.00 13.79 A
ATOM 1880 N ASP A 392 57.264 57.832 52.495 1.00 12.11 A
ATOM 1881 CA ASP A 392 58.008 57.982 51.246 1.00 12.04 A
ATOM 1882 CB ASP A 392 58.327 56.598 50.669 1.00 12.06 A
ATOM 1883 CG ASP A 392 59.425 56.629 49.616 1.00 13.56 A
ATOM 1884 OD1 ASP A 392 59.398 55.754 48.724 1.00 17.06 A
ATOM 1885 OD2 ASP A 392 60.318 57.500 49.687 1.00 9.21 A
ATOM 1886 C ASP A 392 57.190 58.789 50.234 1.00 12.05 A
ATOM 1887 0 ASP A 392 57.744 59.526 49.419 1.00 14.41 A
ATOM 1888 N LEU A 393 55.867 58.649 50.298 1.00 11.58 A
ATOM 1889 CA LEU A 393 54.974 59.363 49.386 1.00 13.19 A
ATOM 1890 CB LEU A 393 53.978 58.383 48.755 1.00 15.37 A
ATOM 1891 CG LEU A 393 54.579 57.277 47.883 1.00 17.56 A
ATOM 1892 CD1 LEU A 393 53.466 56.395 47.334 1.00 18.60 A
ATOM 1893 CD2 LEU A 393 55.377 57.896 46.748 1.00 21.35 A
ATOM 1894 C LEU A 393 54.216 60.465 50.120 1.00 12.25 A
ATOM 1895 0 LEU A 393 53.040 60.714 49.845 1.00 15.76 A
ATOM 1896 N SER A 394 54.897 61.136 51.044 1.00 11.49 A
ATOM 1897 CA SER A 394 54.271 62.192 51.830 1.00 11.54 A
ATOM 1898 CB SER A 394 54.760 62.113 53.279 1.00 13.05 A
ATOM 1899 OG SER A 394 56.151 62.373 53.371 1.00 13.21 A
ATOM 1900 C SER A 394 54.470 63.617 51.310 1.00 11.78 A
ATOM 1901 0 SER A 394 53.823 64.540 51.802 1.00 13.70 A
ATOM 1902 N ASN A 395 55.344 63.800 50.322 1.00 12.05 A
ATOM 1903 CA ASN A 395 55.609 65.134 49.782 1.00 12.13 A
ATOM 1904 CB ASN A 395 56.431 65.055 48.487 1.00 13.30 A
ATOM 1905 CG ASN A 395 57.866 64.607 48.710 1.00 14.95 A
ATOM 1906 001 ASN A 395 58.606 64.412 47.746 1.00 15.49 A
ATOM 1907 ND2 ASN A 395 58.269 64.455 49.969 1.00 15.39 A
ATOM 1908 C ASN A 395 54.347 65.957 49.498 1.00 11.87 A
ATOM 1909 0 ASN A 395 54.237 67.099 49.937 1.00 11.96 A
ATOM 1910 N PRO A 396 53.378 65.383 48.765 1.00 12.62 A
ATOM 1911 CD PRO A 396 53.404 64.106 48.035 1.00 14.02 A
ATOM 1912 CA PRO A 396 52.150 66.126 48.452 1.00 12.71 A
ATOM 1913 CB PRO A 396 51.441 65.218 47.448 1.00 14.17 A
ATOM 1914 CG PRO A 396 52.571 64.431 46.833 1.00 14.79 A
ATOM 1915 C PRO A 396 51.251 66.464 49.633 1.00 12.40 A
ATOM 1916 0 PRO A 396 50.334 67.275 49.492 1.00 13.43 A
ATOM 1917 N THR A 397 51.496 65.845 50.785 1.00 13.56 A
ATOM 1918 CA THR A 397 50.681 66.093 51.972 1.00 12.40 A
ATOM 1919 CB THR A 397 50.506 64.813 52.816 1.00 13.47 A
ATOM 1920 OG1 THR A 397 51.730 64.516 53.502 1.00 14.03 A
ATOM 1921 CG2 THR A 397 50.120 63.644 51.935 1.00 13.51 A
ATOM 1922 C THR A 397 51.284 67.163 52.876 1.00 11.50 A
ATOM 1923 0 THR A 397 50.691 67.541 53.884 1.00 12.93 A
ATOM 1924 N LYS A 398 52.469 67.642 52.521 1.00 12.26 A
ATOM 1925 CA LYS A 398 53.142 68.662 53.312 1.00 13.58 A
ATOM 1926 CB LYS A 398 54.661 68.560 53.125 1.00 14.82 A
ATOM 1927 CG LYS A 398 55.246 67.231 53.555 1.00 15.74 A
ATOM 1928 CD LYS A 398 55.033 66.994 55.042 1.00 16.62 A
ATOM 1929 CE LYS A 398 55.539 65.623 55.481 1.00 17.02 A
ATOM 1930 NZ LYS A 398 57.019 65.472 55.402 1.00 15.70 A
ATOM 1931 C LYS A 398 52.689 70.055 52.912 1.00 14.25 A

ATOM 1932 0 LYS A 398 52.058 70.240 51.872 1.00 14.15 A
ATOM 1933 N SER A 399 53.022 71.035 53.743 1.00 15.69 A
ATOM 1934 CA SER A 399 52.673 72.418 53.454 1.00 17.71 A
ATOM 1935 CB SER A 399 53.396 73.346 54.431 1.00 20.09 A
ATOM 1936 OG SER A 399 52.806 74.634 54.445 1.00 26.34 A
ATOM 1937 C SER A 399 53.136 72.676 52.020 1.00 16.54 A
ATOM 1938 0 SER A 399 54.171 72.164 51.598 1.00 16.94 A
ATOM 1939 N LEU A 400 52.371 73.462 51.271 1.00 16.60 A
ATOM 1940 CA LEU A 400 52.700 73.741 49.879 1.00 16.02 A
ATOM 1941 CB LEU A 400 51.666 74.702 49.282 1.00 16.60 A
ATOM 1942 CG LEU A 400 51.808 74.998 47.785 1.00 17.23 A
ATOM 1943 CD1 LEU A 400 51.687 73.715 46.978 1.00 18.46 A
ATOM 1944 CD2 LEU A 400 50.734 75.987 47.367 1.00 16.33 A
ATOM 1945 C LEU A 400 54.112 74.271 49.615 1.00 15.55 A
ATOM 1946 0 LEU A 400 54.708 73.933 48.589 1.00 16.70 A
ATOM 1947 N GLU A 401 54.658 75.082 50.520 1.00 15.65 A
ATOM 1948 CA GLU A 401 56.001 75.618 50.302 1.00 16.97 A
ATOM 1949 CB GLU A 401 56.358 76.679 51.355 1.00 19.54 A
ATOM 1950 CG GLU A 401 56.793 76.125 52.699 1.00 21.98 A
ATOM 1951 CD GLU A 401 55.632 75.885 53.636 1.00 24.19 A
ATOM 1952 OE1 GLU A 401 54.513 75.664 53.137 1.00 26.88 A
ATOM 1953 OE2 GLU A 401 55.842 75.906 54.870 1.00 26.57 A
ATOM 1954 C GLU A 401 57.044 74.499 50.311 1.00 15.71 A
ATOM 1955 0 GLU A 401 58.086 74.607 49.661 1.00 17.68 A
ATOM 1956 N LEU A 402 56.759 73.424 51.041 1.00 14.61 A
ATOM 1957 CA LEU A 402 57.677 72.293 51.106 1.00 14.28 A
ATOM 1958 CB LEU A 402 57.445 71.482 52.385 1.00 15.31 A
ATOM 1959 CG LEU A 402 57.764 72.187 53.708 1.00 18.56 A
ATOM 1960 CD1 LEU A 402 57.346 71.313 54.887 1.00 20.28 A
ATOM 1961 CD2 LEU A 402 59.257 72.492 53.761 1.00 22.05 A
ATOM 1962 C LEU A 402 57.455 71.406 49.889 1.00 12.37 A
ATOM 1963 0 LEU A 402 58.402 71.000 49.217 1.00 13.91 A
ATOM 1964 N TYR A 403 56.189 71.120 49.601 1.00 13.12 A
ATOM 1965 CA TYR A 403 55.833 70.280 48.467 1.00 13.06 A
ATOM 1966 CB TYR A 403 54.308 70.151 48.413 1.00 11.94 A
ATOM 1967 CG TYR A 403 53.740 69.369 47.248 1.00 12.99 A
ATOM 1968 CD1 TYR A 403 54.446 68.332 46.648 1.00 12.79 A
ATOM 1969 CE1 TYR A 403 53.885 67.584 45.618 1.00 14.80 A
ATOM 1970 CD2 TYR A 403 52.460 69.639 46.787 1.00 13.78 A
ATOM 1971 CE2 TYR A 403 51.892 68.901 45.769 1.00 14.19 A
ATOM 1972 CZ TYR A 403 52.604 67.877 45.187 1.00 14.54 A
ATOM 1973 OH TYR A 403 52.023 67.143 44.171 1.00 16.68 A
ATOM 1974 C TYR A 403 56.402 70.846 47,162 1.00 12.98 A
ATOM 1975 0 TYR A 403 56.929 70.098 46.333 1.00 15.47 A
ATOM 1976 N ARG A 404 56.310 72.164 46.992 1.00 13.53 A
ATOM 1977 CA ARG A 404 56.838 72.832 45.802 1.00 14.31 A
ATOM 1978 CB ARG A 404 56.656 74.350 45.923 1.00 17.96 A
ATOM 1979 CG ARG A 404 55.242 74.858 45.681 1.00 22.47 A
ATOM 1980 CD ARG A 404 54.940 75.064 44.198 1.00 24.43 A
ATOM 1981 NE ARG A 404 55.787 76.089 43.587 1.00 27.85 A
ATOM 1982 CZ ARG A 404 57.029 75.886 43.153 1.00 28.36 A
ATOM 1983 NH1 ARG A 404 57.589 74.687 43.251 1.00 31.22 A
ATOM 1984 NH2 ARG A 404 57.717 76.887 42.623 1.00 29.23 A
ATOM 1985 C ARG A 404 58.328 72.533 45.611 1.00 13.28 A
ATOM 1986 0 ARG A 404 58.783 72.298 44.495 1.00 14.00 A
ATOM 1987 N GLN A 405 59.084 72.563 46.704 1.00 12.67 A
ATOM 1988 CA GLN A 405 60.516 72.298 46.639 1.00 14.09 A
ATOM 1989 CB GLN A 405 61.190 72.704 47.955 1.00 16.67 A
ATOM 1990 CG GLN A 405 61.106 74.202 48.232 1.00 19.03 A
ATOM 1991 CD GLN A 405 61.787 74.606 49.527 1.00 19.85 A
ATOM 1992 OE1 GLN A 405 63.013 74.578 49.632 1.00 21.13 A
ATOM 1993 NE2 GLN A 405 60.991 74.977 50.521 1.00 21.41 A
ATOM 1994 C GLN A 405 60.811 70.837 46.318 1.00 12.92 A
ATOM 1995 0 GLN A 405 61.751 70.535 45.573 1.00 14.19 A
ATOM 1996 N TRP A 406 60.018 69.928 46.875 1.00 11.59 A
ATOM 1997 CA TRP A 406 60.218 68.513 46.603 1.00 11.37 A
ATOM 1998 CB TRP A 406 59.309 67.649 47.481 1.00 10.72 A

ATOM 1999 CG TRP A 406 59.683 67.635 48.931 1.00 11.12 A
ATOM 2000 CD2 TRP A 406 60.949 67.261 49.497 1.00 13.39 A
ATOM 2001 CE2 TRP A 406 60.839 67.402 50.894 1.00 13.47 A
ATOM 2002 CE3 TRP A 406 62.161 66.821 48.958 1.00 13.67 A
ATOM 2003 CD1 TRP A 406 58.882 67.976 49.980 1.00 11.18 A
ATOM 2004 NE1 TRP A 406 59.568 67.839 51.166 1.00 12.80 A
ATOM 2005 CZ2 TRP A 406 61.895 67.119 51.759 1.00 14.28 A
ATOM 2006 CZ3 TRP A 406 63.210 66.540 49.819 1.00 14.66 A
ATOM 2007 CH2 TRP A 406 63.070 66.691 51.204 1.00 12.75 A
ATOM 2008 C TRP A 406 59.922 68.224 45.137 1.00 11.97 A
ATOM 2009 0 TRP A 406 60.637 67.460 44.497 1.00 14.63 A
ATOM 2010 N THR A 407 58.867 68.840 44.610 1.00 12.63 A
ATOM 2011 CA THR A 407 58.479 68.635 43.217 1.00 13.93 A
ATOM 2012 CB THR A 407 57.145 69.366 42.908 1.00 14.39 A
ATOM 2013 OG1 THR A 407 56.092 68.785 43.686 1.00 17.39 A
ATOM 2014 CG2 THR A 407 56.790 69.243 41.439 1.00 16.94 A
ATOM 2015 C THR A 407 59.562 69.109 42.254 1.00 13.54 A
ATOM 2016 0 THR A 407 59.867 68.438 41.265 1.00 14.07 A
ATOM 2017 N ASP A 408 60.156 70.258 42.543 1.00 15.33 A
ATOM 2018 CA ASP A 408 61.213 70.778 41.683 1.00 16.70 A
ATOM 2019 CB ASP A 408 61.642 72.171 42.148 1.00 20.83 A
ATOM 2020 CG ASP A 408 60.554 73.211 41.937 1.00 27.28 A
ATOM 2021 OD1 ASP A 408 60.054 73.320 40.798 1.00 30.78 A
ATOM 2022 OD2 ASP A 408 60.196 73.918 42.903 1.00 31.50 A
ATOM 2023 C ASP A 408 62.406 69.826 41.675 1.00 15.96 A
ATOM 2024 0 ASP A 408 63.041 69.628 40.641 1.00 17.76 A
ATOM 2025 N ARG A 409 62.694 69.225 42.826 1.00 14.51 A
ATOM 2026 CA ARG A 409 63.806 68.287 42.926 1.00 13.79 A
ATOM 2027 CB ARG A 409 64.131 67.991 44.391 1.00 14.84 A
ATOM 2028 CG ARG A 409 64.785 69.160 45.097 1.00 15.68 A
ATOM 2029 CD ARG A 409 65.115 68.849 46.538 1.00 17.95 A
ATOM 2030 NE ARG A 409 65.868 69.949 47.134 1.00 18.15 A
ATOM 2031 CZ ARG A 409 66.935 69.786 47.904 1.00 18.97 A
ATOM 2032 NH1 ARG A 409 67.373 68.563 48.174 1.00 19.30 A
ATOM 2033 NH2 ARG A 409 67.574 70.843 48.392 1.00 21.46 A
ATOM 2034 C ARG A 409 63.544 66.984 42.191 1.00 12.51 A
ATOM 2035 0 ARG A 409 64.435 66.456 41.530 1.00 13.34 A
ATOM 2036 N ILE A 410 62.330 66.455 42.294 1.00 12.18 A
ATOM 2037 CA ILE A 410 62.047 65.209 41.607 1.00 14.64 A
ATOM 2038 CB ILE A 410 60.692 64.609 42.052 1.00 17.58 A
ATOM 2039 CG2 ILE A 410 59.542 65.387 41.449 1.00 20.89 A
ATOM 2040 CG1 ILE A 410 60.619 63.145 41.626 1.00 19.91 A
ATOM 2041 CD1 ILE A 410 61.793 62.313 42.112 1.00 22.03 A
ATOM 2042 C ILE A 410 62.058 65.457 40.098 1.00 13.55 A
ATOM 2043 0 ILE A 410 62.538 64.627 39.331 1.00 13.62 A
ATOM 2044 N MET A 411 61.560 66.617 39.681 1.00 13.62 A
ATOM 2045 CA MET A 411 61.543 66.951 38.261 1.00 15.59 A
ATOM 2046 CB MET A 411 60.853 68.299 38.030 1.00 20.43 A
ATOM 2047 CG MET A 411 59.342 68.238 38.164 1.00 29.99 A
ATOM 2048 SD MET A 411 58.603 66.797 37.357 1.00 33.22 A
ATOM 2049 CE MET A 411 58.946 67.148 35.632 1.00 34.66 A
ATOM 2050 C MET A 411 62.976 67.017 37.751 1.00 14.15 A
ATOM 2051 0 MET A 411 63.280 66.517 36.669 1.00 14.88 A
ATOM 2052 N GLU A 412 63.862 67.626 38.538 1.00 13.96 A
ATOM 2053 CA GLU A 412 65.261 67.732 38.134 1.00 15.05 A
ATOM 2054 CB GLU A 412 66.058 68.567 39.137 1.00 17.37 A
ATOM 2055 CG GLU A 412 67.528 68.746 38.765 1.00 22.75 A
ATOM 2056 CD GLU A 412 67.723 69.271 37.350 1.00 28.08 A
ATOM 2057 OE1 GLU A 412 67.911 68.451 36.418 1.00 29.97 A
ATOM 2058 OE2 GLU A 412 67.680 70.510 37.167 1.00 31.57 A
ATOM 2059 C GLU A 412 65.892 66.351 37.999 1.00 14.34 A
ATOM 2060 0 GLU A 412 66.659 66.107 37.069 1.00 15.05 A
ATOM 2061 N GLU A 413 65.567 65.441 38.913 1.00 14.81 A
ATOM 2062 CA GLU A 413 66.133 64.103 38.829 1.00 15.36 A
ATOM 2063 CB GLU A 413 65.848 63.303 40.105 1.00 16.73 A
ATOM 2064 CG GLU A 413 66.713 62.050 40.208 1.00 17.93 A
ATOM 2065 CD GLU A 413 66.647 61.385 41.565 1.00 17.74 A

ATOM 2066 OE1 GLU A 413 66.896 62.075 42.574 1.00 15.92 A
ATOM 2067 OE2 GLU A 413 66.365 60.170 41.616 1.00 18.42 A
ATOM 2068 C GLU A 413 65.576 63.383 37.598 1.00 14.66 A
ATOM 2069 0 GLU A 413 66.294 62.632 36.940 1.00 15.21 A
ATOM 2070 N PHE A 414 64.304 63.623 37.283 1.00 14.54 A
ATOM 2071 CA PHE A 414 63.681 63.017 36.102 1.00 15.60 A
ATOM 2072 CB PHE A 414 62.245 63.520 35.913 1.00 16.81 A
ATOM 2073 CG PHE A 414 61.180 62.567 36.377 1.00 22.22 A
ATOM 2074 CDl PHE A 414 60.802 62.514 37.709 1.00 23.33 A
ATOM 2075 CD2 PHE A 414 60.512 61.765 35.465 1.00 22.65 A
ATOM 2076 CE1 PHE A 414 59.772 61.682 38.122 1.00 23.93 A
ATOM 2077 CE2 PHE A 414 59.482 60.929 35.871 1.00 24.24 A
ATOM 2078 CZ PHE A 414 59.110 60.889 37.200 1.00 23.76 A
ATOM 2079 C PHE A 414 64.476 63.438 34.868 1.00 15.94 A
ATOM 2080 0 PHE A 414 64.818 62.616 34.016 1.00 16.80 A
ATOM 2081 N PHE A 415 64.747 64.737 34.776 1.00 15.83 A
ATOM 2082 CA PHE A 415 65.496 65.284 33.649 1.00 18.64 A
ATOM 2083 CB PHE A 415 65.536 66.815 33.711 1.00 19.60 A
ATOM 2084 CG PHE A 415 64.189 67.462 33.589 1.00 22.12 A
ATOM 2085 CD1 PHE A 415 63.137 66.797 32.983 1.00 22.28 A
ATOM 2086 CD2 PHE A 415 63.979 68.749 34.067 1.00 24.06 A
ATOM 2087 CE1 PHE A 415 61.897 67.400 32.854 1.00 23.90 A
ATOM 2088 CE2 PHE A 415 62.743 69.358 33.940 1.00 21.91 A
ATOM 2089 CZ PHE A 415 61.699 68.681 33.332 1.00 23.33 A
ATOM 2090 C PHE A 415 66.916 64.747 33.616 1.00 19.13 A
ATOM 2091 0 PHE A 415 67.486 64.570 32.537 1.00 20.54 A
ATOM 2092 N GLN A 416 67.491 64.502 34.792 1.00 18.16 A
ATOM 2093 CA GLN A 416 68.846 63.965 34.876 1.00 18.89 A
ATOM 2094 CB GLN A 416 69.304 63.810 36.333 1.00 19.84 A
ATOM 2095 CG GLN A 416 69.381 65.098 37.147 1.00 24.89 A
ATOM 2096 CD GLN A 416 70.277 64.963 38.374 1.00 25.70 A
ATOM 2097 OE1 GLN A 416 71.455 65.328 38.335 1.00 30.88 A
ATOM 2098 NE2 GLN A 416 69.729 64.419 39.462 1.00 21.09 A
ATOM 2099 C GLN A 416 68.866 62.594 34.211 1.00 17.56 A
ATOM 2100 0 GLN A 416 69.767 62.283 33.435 1.00 17.86 A
ATOM 2101 N GLN A 417 67.876 61.765 34.525 1.00 15.75 A
ATOM 2102 CA GLN A 417 67.818 60.434 33.935 1.00 14.60 A
ATOM 2103 CB GLN A 417 66.685 59.603 34.546 1.00 13.97 A
ATOM 2104 CG GLN A 417 66.639 58.182 33.995 1.00 14.64 A
ATOM 2105 CD GLN A 417 65.635 57.293 34.700 1.00 13.58 A
ATOM 2106 OE1 GLN A 417 65.527 57.313 35.925 1.00 16.51 A
ATOM 2107 NE2 GLN A 417 64.903 56.495 33.931 1.00 16.85 A
ATOM 2108 C GLN A 417 67.609 60.557 32.432 1.00 15.37 A
ATOM 2109 0 GLN A 417 68.182 59.799 31.651 1.00 17.14 A
ATOM 2110 N GLY A 418 66.781 61.515 32.030 1.00 15.76 A
ATOM 2111 CA GLY A 418 66.536 61.715 30.614 1.00 16.90 A
ATOM 2112 C GLY A 418 67.838 62.025 29.898 1.00 16.88 A
ATOM 2113 0 GLY A 418 68.061 61.566 28.776 1.00 19.30 A
ATOM 2114 N ASP A 419 68.703 62.798 30.545 1.00 17.22 A
ATOM 2115 CA ASP A 419 69.987 63.141 29.947 1.00 18.10 A
ATOM 2116 CB ASP A 419 70.727 64.178 30.795 1.00 22.23 A
ATOM 2117 CG ASP A 419 70.039 65.535 30.792 1.00 26.68 A
ATOM 2118 001 ASP A 419 69.594 65.977 29.713 1.00 29.27 A
ATOM 2119 OD2 ASP A 419 69.954 66.167 31.865 1.00 29.97 A
ATOM 2120 C ASP A 419 70.847 61.889 29.799 1.00 17.38 A
ATOM 2121 0 ASP A 419 71.489 61.698 28.770 1.00 17.54 A
ATOM 2122 N LYS A 420 70.850 61.033 30.818 1.00 16.19 A
ATOM 2123 CA LYS A 420 71.647 59.809 30.763 1.00 15.05 A
ATOM 2124 CB LYS A 420 71.631 59.084 32.116 1.00 14.57 A
ATOM 2125 CG LYS A 420 72.204 59.882 33.283 1.00 17.19 A
ATOM 2126 CD LYS A 420 72.174 59.057 34.569 1.00 18.24 A
ATOM 2127 CE LYS A 420 72.356 59.928 35.807 1.00 22.24 A
ATOM 2128 NZ LYS A 420 73.628 60.690 35.777 1.00 24.88 A
ATOM 2129 C LYS A 420 71.142 58.862 29.680 1.00 13.72 A
ATOM 2130 0 LYS A 420 71.937 58.226 28.982 1.00 15.98 A
ATOM 2131 N GLU A 421 69.825 58.767 29.530 1.00 13.82 A
ATOM 2132 CA GLU A 421 69.262 57.879 28.522 1.00 15.82 A

ATOM 2133 CB GLU A 421 67.734 57.855 28.630 1.00 16.66 A
ATOM 2134 CG GLU A 421 67.254 57.462 30.023 1.00 18.54 A
ATOM 2135 CD GLU A 421 65.744 57.420 30.150 1.00 22.50 A
ATOM 2136 OE1 GLU A 421 65.068 58.272 29.531 1.00 25.22 A
ATOM 2137 OE2 GLU A 421 65.234 56.542 30.883 1.00 23.66 A
ATOM 2138 C GLU A 421 69.712 58.328 27.133 1.00 16.59 A
ATOM 2139 0 GLU A 421 70.044 57.499 26.284 1.00 18.60 A
ATOM 2140 N ARG A 422 69.744 59.639 26.912 1.00 18.06 A
ATOM 2141 CA ARG A 422 70.187 60.169 25.625 1.00 19.52 A
ATOM 2142 CB ARG A 422 69.891 61.666 25.531 1.00 21.06 A
ATOM 2143 CG ARG A 422 70.514 62.339 24.316 1.00 24.66 A
ATOM 2144 CD ARG A 422 71.685 63.207 24.731 1.00 24.97 A
ATOM 2145 NE ARG A 422 71.242 64.306 25.584 1.00 25.82 A
ATOM 2146 CZ ARG A 422 72.052 65.087 26.291 1.00 24.05 A
ATOM 2147 NH1 ARG A 422 73.368 64.901 26.256 1.00 22.03 A
ATOM 2148 NH2 ARG A 422 71.541 66.053 27.039 1.00 26.82 A
ATOM 2149 C ARG A 422 71.683 59.928 25.440 1.00 18.68 A
ATOM 2150 0 ARG A 422 72.124 59.477 24.379 1.00 21.26 A
ATOM 2151 N GLU A 423 72.462 60.221 26.477 1.00 17.69 A
ATOM 2152 CA GLU A 423 73.909 60.034 26.422 1.00 16.75 A
ATOM 2153 CB GLU A 423 74.547 60.401 27.770 1.00 18.92 A
ATOM 2154 CG GLU A 423 74.351 61.853 28.183 1.00 20.70 A
ATOM 2155 CD GLU A 423 74.874 62.138 29.578 1.00 22.80 A
ATOM 2156 OEl GLU A 423 74.815 61.225 30.429 1.00 23.73 A
ATOM 2157 OE2 GLU A 423 75.332 63.274 29.831 1.00 24.67 A
ATOM 2158 C GLU A 423 74.280 58.595 26.073 1.00 16.25 A
ATOM 2159 0 GLU A 423 75.246 58.356 25.351 1.00 16.81 A
ATOM 2160 N ARG A 424 73.506 57.642 26.582 1.00 15.35 A
ATOM 2161 CA ARG A 424 73.785 56.230 26.351 1.00 16.43 A
ATOM 2162 CB ARG A 424 73.517 55.443 27.634 1.00 15.39 A
ATOM 2163 CG ARG A 424 74.309 55.977 28.812 1.00 16.51 A
ATOM 2164 CD ARG A 424 74.112 55.142 30.052 1.00 16.22 A
ATOM 2165 NE ARG A 424 74.601 55.843 31.236 1.00 16.27 A
ATOM 2166 CZ ARG A 424 74.771 55.273 32.423 1.00 15.17 A
ATOM 2167 NH1 ARG A 424 74.501 53.985 32.587 1.00 16.11 A
ATOM 2168 NH2 ARG A 424 75.184 56.000 33.452 1.00 13.90 A
ATOM 2169 C ARG A 424 73.015 55.610 25.192 1.00 17.91 A
ATOM 2170 0 ARG A 424 73.048 54.392 25.002 1.00 19.16 A
ATOM 2171 N GLY A 425 72.323 56.451 24.431 1.00 20.78 A
ATOM 2172 CA GLY A 425 71.554 55.981 23.288 1.00 22.94 A
ATOM 2173 C GLY A 425 70.571 54.867 23.598 1.00 24.03 A
ATOM 2174 0 GLY A 425 70.403 53.943 22.800 1.00 24.64 A
ATOM 2175 N MET A 426 69.908 54.958 24.748 1.00 22.23 A
ATOM 2176 CA MET A 426 68.943 53.945 25.174 1.00 23.17 A
ATOM 2177 C MET A 426 67.724 53.866 24.256 1.00 23.48 A
ATOM 2178 0 MET A 426 67.153 54.886 23.871 1.00 24.45 A
ATOM 2179 CB MET A 426 68.608 54.189 26.638 1.00 22.64 A
ATOM 2180 CG MET A 426 69.821 54.381 27.547 1.00 20.00 A
ATOM 2181 SD MET A 426 69.405 54.178 29.293 1.00 20.00 A
ATOM 2182 CE MET A 426 67.864 53.279 29.170 1.00 20.00 A
ATOM 2183 N GLU A 427 67.471 52.497 23.897 1.00 23.43 A
ATOM 2184 CA GLU A 427 66.410 52.171 22.949 1.00 24.49 A
ATOM 2185 C GLU A 427 65.011 52.254 23.557 1.00 25.24 A
ATOM 2186 0 GLU A 427 64.314 51.245 23.673 1.00 26.50 A
ATOM 2187 CB GLU A 427 66.611 50.765 22.383 1.00 24.34 A
ATOM 2188 CG GLU A 427 67.924 50.576 21.644 1.00 20.00 A
ATOM 2189 CD GLU A 427 68.088 49.171 21.098 1.00 20.00 A
ATOM 2190 OE1 GLU A 427 67.134 48.661 20.475 1.00 20.00 A
ATOM 2191 OE2 GLU A 427 69.171 48.581 21.293 1.00 20.00 A
ATOM 2192 N ILE A 428 64.608 53.461 23.939 1.00 25.29 A
ATOM 2193 CA ILE A 428 63.290 53.695 24.523 1.00 25.05 A
ATOM 2194 CB ILE A 428 63.295 53.512 26.060 1.00 24.97 A
ATOM 2195 CG2 ILE A 428 63.583 52.064 26.416 1.00 24.93 A
ATOM 2196 CG1 ILE A 428 64.327 54.444 26.696 1.00 22.21 A
ATOM 2197 CD1 ILE A 428 64.427 54.289 28.202 1.00 24.76 A
ATOM 2198 C ILE A 428 62.845 55.118 24.211 1.00 26.65 A
ATOM 2199 0 ILE A 428 63.651 55.943 23.780 1.00 26.59 A

ATOM 2200 N ALA A 437 56.662 70.439 30.126 1.00 27.00 A
ATOM 2201 CA ALA A 437 56.829 69.117 30.715 1.00 26.16 A
ATOM 2202 CB ALA A 437 58.028 69.115 31.650 1.00 24.91 A
ATOM 2203 C ALA A 437 55.572 68.702 31.472 1.00 26.13 A
ATOM 2204 0 ALA A 437 55.456 67.560 31.927 1.00 27.05 A
ATOM 2205 N SER A 438 54.634 69.637 31.601 1.00 25.02 A
ATOM 2206 CA SER A 438 53.380 69.387 32.298 1.00 24.02 A
ATOM 2207 CB SER A 438 52.609 68.256 31.615 1.00 24.69 A
ATOM 2208 OG SER A 438 52.322 68.578 30.264 1.00 27.74 A
ATOM 2209 C SER A 438 53.611 69.029 33.760 1.00 22.54 A
ATOM 2210 0 SER A 438 53.019 68.080 34.270 1.00 22.65 A
ATOM 2211 N VAL A 439 54.467 69.793 34.434 1.00 21.81 A
ATOM 2212 CA VAL A 439 54.764 69.543 35.840 1.00 22.02 A
ATOM 2213 CB VAL A 439 55.733 70.601 36.412 1.00 22.20 A
ATOM 2214 CG1 VAL A 439 55.988 70.328 37.886 1.00 23.44 A
ATOM 2215 CG2 VAL A 439 57.030 70.589 35.636 1.00 23.59 A
ATOM 2216 C VAL A 439 53.496 69.560 36.683 1.00 21.35 A
ATOM 2217 0 VAL A 439 53.251 68.646 37.472 1.00 20.99 A
ATOM 2218 N GLU A 440 52.691 70.604 36.508 1.00 21.61 A
ATOM 2219 CA GLU A 440 51.450 70.745 37.262 1.00 21.11 A
ATOM 2220 CB GLU A 440 50.753 72.060 36.897 1.00 22.04 A
ATOM 2221 CG GLU A 440 51.607 73.293 37.145 1.00 23.80 A
ATOM 2222 CD GLU A 440 52.283 73.804 35.886 1.00 27.53 A
ATOM 2223 OE1 GLU A 440 52.820 72.981 35.115 1.00 26.96 A
ATOM 2224 OE2 GLU A 440 52.280 75.037 35.670 1.00 31.26 A
ATOM 2225 C GLU A 440 50.499 69.580 37.016 1.00 21.18 A
ATOM 2226 0 GLU A 440 50.033 68.940 37.959 1.00 21.38 A
ATOM 2227 N LYS A 441 50.212 69.307 35.747 1.00 20.82 A
ATOM 2228 CA LYS A 441 49.312 68.217 35.400 1.00 21.66 A
ATOM 2229 CB LYS A 441 49.110 68.165 33.881 1.00 22.41 A
ATOM 2230 CG LYS A 441 48.183 67.057 33.409 1.00 24.36 A
ATOM 2231 CD LYS A 441 47.424 67.480 32.161 1.00 24.61 A
ATOM 2232 CE LYS A 441 48.344 68.123 31.133 1.00 25.36 A
ATOM 2233 NZ LYS A 441 49.377 67.187 30.605 1.00 27.46 A
ATOM 2234 C LYS A 441 49.853 66.887 35.912 1.00 21.45 A
ATOM 2235 0 LYS A 441 49.088 66.003 36.305 1.00 22.14 A
ATOM 2236 N SER A 442 51.174 66.753 35.915 1.00 21.15 A
ATOM 2237 CA SER A 442 51.806 65.532 36.388 1.00 20.90 A
ATOM 2238 CB SER A 442 53.318 65.590 36.161 1.00 21.81 A
ATOM 2239 OG SER A 442 53.946 66.320 37.200 1.00 27.97 A
ATOM 2240 C SER A 442 51.514 65.292 37.863 1.00 19.02 A
ATOM 2241 0 SER A 442 51.263 64.161 38.270 1.00 19.71 A
ATOM 2242 N GLN A 443 51.541 66.351 38.663 1.00 19.25 A
ATOM 2243 CA GLN A 443 51.272 66.208 40.089 1.00 17.82 A
ATOM 2244 CB GLN A 443 51.636 67.492 40.836 1.00 19.26 A
ATOM 2245 CG GLN A 443 53.130 67.760 40.874 1.00 17.81 A
ATOM 2246 CD GLN A 443 53.924 66.544 41.316 1.00 18.23 A
ATOM 2247 OE1 GLN A 443 53.732 66.024 42.416 1.00 19.26 A
ATOM 2248 NE2 GLN A 443 54.824 66.084 40.455 1.00 21.49 A
ATOM 2249 C GLN A 443 49.816 65.844 40.350 1.00 17.57 A
ATOM 2250 0 GLN A 443 49.514 65.045 41.233 1.00 18.11 A
ATOM 2251 N VAL A 444 48.904 66.438 39.589 1.00 17.31 A
ATOM 2252 CA VAL A 444 47.497 66.119 39.763 1.00 17.48 A
ATOM 2253 CB VAL A 444 46.611 66.953 38.811 1.00 17.80 A
ATOM 2254 CG1 VAL A 444 45.149 66.553 38.975 1.00 18.89 A
ATOM 2255 CG2 VAL A 444 46.791 68.434 39.109 1.00 18.60 A
ATOM 2256 C VAL A 444 47.309 64.633 39.470 1.00 17.11 A
ATOM 2257 0 VAL A 444 46.638 63.919 40.218 1.00 20.12 A
ATOM 2258 N GLY A 445 47.920 64.174 38.383 1.00 17.96 A
ATOM 2259 CA GLY A 445 47.814 62.774 38.007 1.00 18.54 A
ATOM 2260 C GLY A 445 48.439 61.850 39.035 1.00 18.10 A
ATOM 2261 0 GLY A 445 47.877 60.810 39.379 1.00 19.74 A
ATOM 2262 N PHE A 446 49.611 62.232 39.528 1.00 18.18 A
ATOM 2263 CA PHE A 446 50.319 61.438 40.525 1.00 17.21 A
ATOM 2264 CB PHE A 446 51.666 62.099 40.850 1.00 16.65 A
ATOM 2265 CG PHE A 446 52.487 61.352 41.868 1.00 16.58 A
ATOM 2266 CD1 PHE A 446 52.708 59.991 41.736 1.00 16.98 A

ATOM 2267 CD2 PHE A 446 53.053 62.019 42.944 1.00 17.14 A
ATOM 2268 CEl PHE A 446 53.481 59.302 42.661 1.00 17.13 A
ATOM 2269 CE2 PHE A 446 53.828 61.336 43.874 1.00 18.97 A
ATOM 2270 CZ PHE A 446 54.040 59.977 43.730 1.00 17.95 A
ATOM 2271 C PHE A 446 49.475 61.305 41.786 1.00 16.79 A
ATOM 2272 0 PHE A 446 49.341 60.221 42.353 1.00 15.56 A
ATOM 2273 N ILE A 447 48.897 62.416 42.223 1.00 16.42 A
ATOM 2274 CA ILE A 447 48.069 62.415 43.416 1.00 16.48 A
ATOM 2275 CB ILE A 447 47.676 63.859 43.808 1.00 17.35 A
ATOM 2276 CG2 ILE A 447 46.676 63.845 44.950 1.00 16.77 A
ATOM 2277 CG1 ILE A 447 48.931 64.645 44.199 1.00 17.64 A
ATOM 2278 CD1 ILE A 447 48.667 66.095 44.531 1.00 19.00 A
ATOM 2279 C ILE A 447 46.809 61.566 43.234 1.00 15.76 A
ATOM 2280 0 ILE A 447 46.490 60.726 44.071 1.00 16.55 A
ATOM 2281 N ASP A 448 46.094 61.771 42.133 1.00 16.95 A
ATOM 2282 CA ASP A 448 44.875 61.003 41.904 1.00 18.92 A
ATOM 2283 CB ASP A 448 44.108 61.558 40.700 1.00 20.70 A
ATOM 2284 CG ASP A 448 43.544 62.943 40.951 1.00 23.82 A
ATOM 2285 OD1 ASP A 448 43.289 63.285 42.125 1.00 24.89 A
ATOM 2286 OD2 ASP A 448 43.333 63.686 39.970 1.00 25.78 A
ATOM 2287 C ASP A 448 45.113 59.511 41.692 1.00 19.53 A
ATOM 2288 0 ASP A 448 44.313 58.679 42.124 1.00 22.34 A
ATOM 2289 N TYR A 449 46.216 59.175 41.036 1.00 19.46 A
ATOM 2290 CA TYR A 449 46.535 57.786 40.734 1.00 20.81 A
ATOM 2291 CB TYR A 449 47.400 57.729 39.472 1.00 22.34 A
ATOM 2292 CG TYR A 449 47.595 56.337 38.908 1.00 25.79 A
ATOM 2293 CD1 TYR A 449 46.512 55.591 38.457 1.00 25.80 A
ATOM 2294 CE1 TYR A 449 46.686 54.326 37.926 1.00 27.69 A
ATOM 2295 CD2 TYR A 449 48.863 55.776 38.812 1.00 25.27 A
ATOM 2296 CE2 TYR A 449 49.047 54.509 38.281 1.00 27.63 A
ATOM 2297 CZ TYR A 449 47.955 53.790 37.840 1.00 27.04 A
ATOM 2298 OH TYR A 449 48.125 52.527 37.319 1.00 31.23 A
ATOM 2299 C TYR A 449 47.228 56.999 41.844 1.00 20.67 A
ATOM 2300 0 TYR A 449 46.946 55.813 42.032 1.00 20.46 A
ATOM 2301 N ILE A 450 48.119 57.653 42.585 1.00 19.13 A
ATOM 2302 CA ILE A 450 48.882 56.966 43.626 1.00 17.88 A
ATOM 2303 CB ILE A 450 50.393 56.992 43.294 1.00 19.39 A
ATOM 2304 CG2 ILE A 450 51.168 56.209 44.346 1.00 19.89 A
ATOM 2305 CG1 ILE A 450 50.649 56.412 41.902 1.00 20.97 A
ATOM 2306 CD1 ILE A 450 50.374 54.937 41.800 1.00 22.24 A
ATOM 2307 C ILE A 450 48.757 57.493 45.053 1.00 15.95 A
ATOM 2308 0 ILE A 450 48.404 56.759 45.980 1.00 17.81 A
ATOM 2309 N VAL A 451 49.065 58.772 45.225 1.00 15.28 A
ATOM 2310 CA VAL A 451 49.082 59.384 46.544 1.00 14.64 A
ATOM 2311 CB VAL A 451 49.722 60.768 46.456 1.00 13.97 A
ATOM 2312 CG1 VAL A 451 50.050 61.273 47.844 1.00 13.25 A
ATOM 2313 CG2 VAL A 451 50.984 60.689 45.595 1.00 14.44 A
ATOM 2314 C VAL A 451 47.760 59.479 47.292 1.00 14.72 A
ATOM 2315 0 VAL A 451 47.676 59.082 48.455 1.00 15.41 A
ATOM 2316 N HIS A 452 46.730 59.998 46.636 1.00 16.82 A
ATOM 2317 CA HIS A 452 45.429 60.127 47.274 1.00 17.91 A
ATOM 2318 CB HIS A 452 44.468 60.882 46.357 1.00 19.59 A
ATOM 2319 CG HIS A 452 43.125 61.129 46.968 1.00 22.77 A
ATOM 2320 CD2 HIS A 452 41.883 60.776 46.562 1.00 23.93 A
ATOM 2321 ND1 HIS A 452 42.961 61.825 48.146 1.00 25.09 A
ATOM 2322 CE1 HIS A 452 41.674 61.892 48.438 1.00 26.34 A
ATOM 2323 NE2 HIS A 452 40.999 61.263 47.493 1.00 25.97 A
ATOM 2324 C HIS A 452 44.820 58.779 47.680 1.00 17.05 A
ATOM 2325 0 HIS A 452 44.364 58.618 48.809 1.00 19.16 A
ATOM 2326 N PRO A 453 44.803 57.795 46.767 1.00 18.47 A
ATOM 2327 CD PRO A 453 45.173 57.836 45.341 1.00 19.12 A
ATOM 2328 CA PRO A 453 44.230 56.490 47.121 1.00 18.21 A
ATOM 2329 CB PRO A 453 44.498 55.650 45.877 1.00 18.94 A
ATOM 2330 CG PRO A 453 44.411 56.660 44.776 1.00 19.04 A
ATOM 2331 C PRO A 453 44.891 55.900 48.365 1.00 17.61 A
ATOM 2332 0 PRO A 453 44.230 55.324 49.232 1.00 18.57 A
ATOM 2333 N LEU A 454 46.206 56.057 48.444 1.00 16.41 A

ATOM 2334 CA LEU A 454 46.970 55.545 49.567 1.00 16.49 A
ATOM 2335 CB LEU A 454 48.468 55.670 49.272 1.00 16.31 A
ATOM 2336 CG LEU A 454 49.424 55.307 50.410 1.00 18.71 A
ATOM 2337 CD1 LEU A 454 49.218 53.853 50.808 1.00 20.00 A
ATOM 2338 CD2 LEU A 454 50.865 55.547 49.969 1.00 18.02 A
ATOM 2339 C LEU A 454 46.645 56.259 50.876 1.00 15.50 A
ATOM 2340 0 LEU A 454 46.304 55.618 51.871 1.00 16.68 A
ATOM 2341 N TRP A 455 46.749 57.584 50.879 1.00 16.34 A
ATOM 2342 CA TRP A 455 46.483 58.348 52.092 1.00 15.60 A
ATOM 2343 CB TRP A 455 46.938 59.797 51.924 1.00 16.50 A
ATOM 2344 CG TRP A 455 48.416 59.950 52.113 1.00 15.47 A
ATOM 2345 CD2 TRP A 455 49.112 60.002 53.360 1.00 15.27 A
ATOM 2346 CE2 TRP A 455 50.485 60.114 53.068 1.00 14.82 A
ATOM 2347 CE3 TRP A 455 48.706 59.964 54.698 1.00 16.51 A
ATOM 2348 CDl TRP A 455 49.368 60.030 51.138 1.00 15.88 A
ATOM 2349 NE1 TRP A 455 50.616 60.130 51.704 1.00 15.63 A
ATOM 2350 CZ2 TRP A 455 51.456 60.190 54.065 1.00 16.36 A
ATOM 2351 CZ3 TRP A 455 49.673 60.040 55.688 1.00 18.58 A
ATOM 2352 CH2 TRP A 455 51.031 60.152 55.364 1.00 18.21 A
ATOM 2353 C TRP A 455 45.038 58.305 52.554 1.00 16.69 A
ATOM 2354 0 TRP A 455 44.764 58.412 53.747 1.00 18.08 A
ATOM 2355 N GLU A 456 44.118 58.150 51.613 1.00 17.85 A
ATOM 2356 CA GLU A 456 42.707 58.071 51.959 1.00 19.00 A
ATOM 2357 CB GLU A 456 41.861 58.044 50.685 1.00 19.73 A
ATOM 2358 CG GLU A 456 40.366 58.016 50.916 1.00 24.82 A
ATOM 2359 CD GLU A 456 39.591 58.123 49.618 1.00 27.41 A
ATOM 2360 OE1 GLU A 456 39.805 57.270 48.729 1.00 29.58 A
ATOM 2361 OE2 GLU A 456 38.771 59.060 49.486 1.00 31.04 A
ATOM 2362 C GLU A 456 42.490 56.795 52.766 1.00 18.98 A
ATOM 2363 0 GLU A 456 41.706 56.770 53.714 1.00 21.23 A
ATOM 2364 N THR A 457 43.204 55.741 52.389 1.00 19.38 A
ATOM 2365 CA THR A 457 43.091 54.460 53.072 1.00 19.63 A
ATOM 2366 CB THR A 457 43.706 53.338 52.220 1.00 20.45 A
ATOM 2367 OG1 THR A 457 43.095 53.345 50.923 1.00 21.84 A
ATOM 2368 CG2 THR A 457 43.488 51.985 52.878 1.00 21.06 A
ATOM 2369 C THR A 457 43.779 54.517 54.434 1.00 19.60 A
ATOM 2370 0 THR A 457 43.281 53.962 55.413 1.00 20.03 A
ATOM 2371 N TRP A 458 44.926 55.185 54.502 1.00 18.12 A
ATOM 2372 CA TRP A 458 45.631 55.306 55.768 1.00 17.21 A
ATOM 2373 CB TRP A 458 46.987 55.995 55.565 1.00 17.17 A
ATOM 2374 CG TRP A 458 47.695 56.298 56.858 1.00 17.09 A
ATOM 2375 CD2 TRP A 458 48.540 55.408 57.596 1.00 17.34 A
ATOM 2376 CE2 TRP A 458 48.948 56.088 58.762 1.00 17.95 A
ATOM 2377 CE3 TRP A 458 48.990 54.101 57.384 1.00 18.88 A
ATOM 2378 CD1 TRP A 458 47.626 57.449 57.583 1.00 17.40 A
ATOM 2379 NE1 TRP A 458 48.378 57.333 58.732 1.00 16.65 A
ATOM 2380 CZ2 TRP A 458 49.785 55.504 59.714 1.00 18.09 A
ATOM 2381 CZ3 TRP A 458 49.823 53.522 58.331 1.00 19.06 A
ATOM 2382 CH2 TRP A 458 50.210 54.223 59.479 1.00 18.64 A
ATOM 2383 C TRP A 458 44.763 56.117 56.731 1.00 16.76 A
ATOM 2384 0 TRP A 458 44.670 55.805 57.917 1.00 16.18 A
ATOM 2385 N ALA A 459 44.117 57.151 56.203 1.00 17.73 A
ATOM 2386 CA ALA A 459 43.247 58.007 57.000 1.00 18.94 A
ATOM 2387 CB ALA A 459 42.693 59.130 56.136 1.00 19.79 A
ATOM 2388 C ALA A 459 42.102 57.203 57.606 1.00 20.49 A
ATOM 2389 0 ALA A 459 41.685 57.459 58.734 1.00 21.34 A
ATOM 2390 N ASP A 460 41.593 56.230 56.856 1.00 21.63 A
ATOM 2391 CA ASP A 460 40.500 55.402 57.353 1.00 22.89 A
ATOM 2392 CB ASP A 460 39.924 54.523 56.234 1.00 25.00 A
ATOM 2393 CG ASP A 460 39.316 55.328 55.100 1.00 28.11 A
ATOM 2394 OD1 ASP A 460 38.497 56.233 55.375 1.00 30.44 A
ATOM 2395 OD2 ASP A 460 39.648 55.046 53.927 1.00 32.29 A
ATOM 2396 C ASP A 460 40.996 54.506 58.485 1.00 23.03 A
ATOM 2397 0 ASP A 460 40.273 54.241 59.444 1.00 23.84 A
ATOM 2398 N LEU A 461 42.240 54.048 58.372 1.00 21.32 A
ATOM 2399 CA LEU A 461 42.825 53.170 59.376 1.00 20.64 A
ATOM 2400 CB LEU A 461 44.184 52.647 58.900 1.00 19.34 A

ATOM 2401 CG LEU A 461 44.865 51.648 59.840 1.00 20.72 A
ATOM 2402 CD1 LEU A 461 44.074 50.340 59.837 1.00 19.53 A
ATOM 2403 CD2 LEU A 461 46.303 51.403 59.397 1.00 21.88 A
ATOM 2404 C LEU A 461 43.004 53.852 60.725 1.00 20.88 A
ATOM 2405 0 LEU A 461 42.713 53.264 61.763 1.00 21.41 A
ATOM 2406 N VAL A 462 43.481 55.092 60.705 1.00 20.72 A
ATOM 2407 CA VAL A 462 43.726 55.836 61.933 1.00 20.37 A
ATOM 2408 CB VAL A 462 45.153 56.419 61.930 1.00 20.38 A
ATOM 2409 CG1 VAL A 462 46.168 55.299 61.778 1.00 20.34 A
ATOM 2410 CG2 VAL A 462 45.302 57.418 60.792 1,00 19.63 A
ATOM 2411 C VAL A 462 42.746 56.982 62.175 1.00 21.10 A
ATOM 2412 0 VAL A 462 43.007 57.857 62.999 1.00 22.25 A
ATOM 2413 N GLN A 463 41.617 56.975 61.474 1.00 21.86 A
ATOM 2414 CA GLN A 463 40.632 58.042 61.629 1.00 22.64 A
ATOM 2415 CB GLN A 463 39.306 57.654 60.971 1.00 23.80 A
ATOM 2416 CG GLN A 463 38.686 56.379 61.509 1.00 26.52 A
ATOM 2417 CD GLN A 463 37.270 56.171 61.007 1.00 27.52 A
ATOM 2418 OE1 GLN A 463 36.659 55.126 61.246 1.00 31.57 A
ATOM 2419 NE2 GLN A 463 36.735 57.172 60.314 1.00 28.38 A
ATOM 2420 C GLN A 463 40.385 58.405 63.090 1.00 23.04 A
ATOM 2421 0 GLN A 463 40.306 57.533 63.956 1.00 23.18 A
ATOM 2422 N PRO A 464 40.255 59.708 63.378 1.00 23.68 A
ATOM 2423 CD PRO A 464 39.630 60.181 64.629 1.00 23.90 A
ATOM 2424 CA PRO A 464 40.334 60.802 62.405 1.00 23.00 A
ATOM 2425 CB PRO A 464 39.180 61.688 62.828 1.00 23.85 A
ATOM 2426 CG PRO A 464 39.337 61.652 64.325 1.00 24.65 A
ATOM 2427 C PRO A 464 41.665 61.550 62.507 1.00 23.08 A
ATOM 2428 0 PRO A 464 41.746 62.735 62.188 1.00 24.27 A
ATOM 2429 N ASP A 465 42.707 60.855 62.946 1.00 22.95 A
ATOM 2430 CA ASP A 465 44.011 61.477 63.132 1.00 23.05 A
ATOM 2431 CB ASP A 465 44.931 60.512 63.880 1.00 25.13 A
ATOM 2432 CG ASP A 465 44.428 60.205 65.274 1.00 27.12 A
ATOM 2433 OD1 ASP A 465 44.205 61.163 66.046 1.00 30.09 A
ATOM 2434 OD2 ASP A 465 44.253 59.013 65.596 1.00 29.50 A
ATOM 2435 C ASP A 465 44.726 62.008 61.893 1.00 23.27 A
ATOM 2436 0 ASP A 465 45.592 62.873 62.006 1.00 24.33 A
ATOM 2437 N ALA A 466 44.366 61.515 60.715 1.00 21.56 A
ATOM 2438 CA ALA A 466 45.025 61.974 59.497 1.00 21.08 A
ATOM 2439 CB ALA A 466 45.517 60.774 58.691 1.00 19.38 A
ATOM 2440 C ALA A 466 44.151 62.865 58.621 1.00 21.07 A
ATOM 2441 0 ALA A 466 44.452 63.067 57.445 1.00 22.35 A
ATOM 2442 N GLN A 467 43.077 63.409 59.184 1.00 21.16 A
ATOM 2443 CA GLN A 467 42.196 64.266 58.401 1.00 20.54 A
ATOM 2444 CB GLN A 467 40.965 64.668 59.213 1.00 21.53 A
ATOM 2445 CG GLN A 467 39.902 65.358 58.370 1.00 23.82 A
ATOM 2446 CD GLN A 467 39.518 64.538 57.147 1.00 25.04 A
ATOM 2447 OE1 GLN A 467 39.103 63.385 57.265 1.00 27.25 A
ATOM 2448 NE2 GLN A 467 39.657 65.131 55.966 1.00 26.57 A
ATOM 2449 C GLN A 467 42.903 65.520 57.900 1.00 20.69 A
ATOM 2450 0 GLN A 467 42.684 65.944 56.766 1.00 21.24 A
ATOM 2451 N ASP A 468 43.740 66.118 58.744 1.00 20.82 A
ATOM 2452 CA ASP A 468 44.469 67.327 58.370 1.00 21.77 A
ATOM 2453 CB ASP A 468 45.255 67.864 59.566 1.00 23.31 A
ATOM 2454 CG ASP A 468 44.364 68.190 60.747 1.00 27.59 A
ATOM 2455 OD1 ASP A 468 43.435 69.009 60.585 1.00 30.16 A
ATOM 2456 OD2 ASP A 468 44.595 67.628 61.838 1.00 29.99 A
ATOM 2457 C ASP A 468 45.426 67.054 57.213 1.00 20.85 A
ATOM 2458 0 ASP A 468 45.630 67.905 56.347 1.00 21.91 A
ATOM 2459 N ILE A 469 46.011 65.863 57.206 1.00 20.82 A
ATOM 2460 CA ILE A 469 46.929 65.480 56.144 1.00 20.02 A
ATOM 2461 CB ILE A 469 47.619 64.134 56.470 1.00 19.73 A
ATOM 2462 CG2 ILE A 469 48.365 63.616 55.246 1.00 22.81 A
ATOM 2463 CG1 ILE A 469 48.580 64.320 57.649 1.00 21.01 A
ATOM 2464 CD1 ILE A 469 49.276 63.038 58.093 1.00 19.17 A
ATOM 2465 C ILE A 469 46.151 65.368 54.830 1.00 19.76 A
ATOM 2466 0 ILE A 469 46.618 65.817 53.781 1.00 19.64 A
ATOM 2467 N LEU A 470 44.958 64.785 54.898 1.00 19.07 A

ATOM 2468 CA LEU A 470 44.113 64.634 53.718 1.00 19.45 A
ATOM 2469 CB LEU A 470 42.880 63.785 54.035 1.00 20.05 A
ATOM 2470 CG LEU A 470 43.013 62.262 54.008 1.00 20.72 A
ATOM 2471 CD1 LEU A 470 41.642 61.650 54.265 1.00 22.80 A
ATOM 2472 CD2 LEU A 470 43.539 61.806 52.654 1.00 22.28 A
ATOM 2473 C LEU A 470 43.654 65.990 53.190 1.00 18.98 A
ATOM 2474 0 LEU A 470 43.591 66.203 51.977 1.00 18.71 A
ATOM 2475 N ASP A 471 43.325 66.902 54.101 1.00 19.76 A
ATOM 2476 CA ASP A 471 42.873 68.231 53.703 1.00 19.53 A
ATOM 2477 CB ASP A 471 42.417 69.040 54.920 1.00 21.75 A
ATOM 2478 CG ASP A 471 41.141 68.503 55.532 1.00 24.97 A
ATOM 2479 OD1 ASP A 471 40.375 67.823 54.815 1.00 27.27 A
ATOM 2480 OD2 ASP A 471 40.894 68.776 56.725 1.00 27.42 A
ATOM 2481 C ASP A 471 43.964 68.997 52.963 1.00 18.18 A
ATOM 2482 0 ASP A 471 43.693 69.698 51.992 1.00,18.06 A
ATOM 2483 N THR A 472 45.202 68.862 53.424 1.00 17.25 A
ATOM 2484 CA THR A 472 46.316 69.539 52.778 1.00 15.40 A
ATOM 2485 CB THR A 472 47.590 69.443 53.630 1.00 15.27 A
ATOM 2486 001 THR A 472 47.367 70.089 54.888 1.00 17.72 A
ATOM 2487 CG2 THR A 472 48.757 70.115 52.919 1.00 14.24 A
ATOM 2488 C THR A 472 46.586 68.915 51.413 1.00 14.97 A
ATOM 2489 0 THR A 472 46.874 69.620 50.445 1.00 16.05 A
ATOM 2490 N LEU A 473 46.488 67.590 51.339 1.00 15.60 A
ATOM 2491 CA LEU A 473 46.714 66.888 50.084 1.00 15.63 A
ATOM 2492 CB LEU A 473 46.537 65.380 50.273 1.00 16.14 A
ATOM 2493 CG LEU A 473 46.760 64.530 49.020 1.00 15.16 A
ATOM 2494 CDl LEU A 473 48.170 64.754 48.486 1.00 15.86 A
ATOM 2495 CD2 LEU A 473 46.538 63.063 49.349 1.00 18.27 A
ATOM 2496 C LEU A 473 45.722 67.398 49.048 1.00 15.41 A
ATOM 2497 0 LEU A 473 46.086 67.677 47.908 1.00 16.02 A
ATOM 2498 N GLU A 474 44.467 67.525 49.470 1.00 17.29 A
ATOM 2499 CA GLU A 474 43.395 68.006 48.608 1.00 17.12 A
ATOM 2500 CB GLU A 474 42.066 67.936 49.361 1.00 19.50 A
ATOM 2501 CG GLU A 474 40.859 68.470 48.605 1.00 23.04 A
ATOM 2502 CD GLU A 474 40.510 67.650 47.377 1.00 26.60 A
ATOM 2503 OE1 GLU A 474 40.794 66.433 47.364 1.00 28.66 A
ATOM 2504 OE2 GLU A 474 39.932 68.221 46.428 1.00 29.77 A
ATOM 2505 C GLU A 474 43.667 69.438 48.152 1.00 16.00 A
ATOM 2506 0 GLU A 474 43.547 69.749 46.965 1.00 16.27 A
ATOM 2507 N ASP A 475 44.032 70.305 49.094 1.00 16.49 A
ATOM 2508 CA ASP A 475 44.328 71.698 48.767 1.00 16.91 A
ATOM 2509 CB ASP A 475 44.652 72.509 50.027 1.00 18.92 A
ATOM 2510 CG ASP A 475 43.435 72.753 50.904 1.00 22.25 A
ATOM 2511 OD1 ASP A 475 42.295 72.576 50.421 1.00 24.34 A
ATOM 2512 OD2 ASP A 475 43.625 73.137 52.080 1.00 25.35 A
ATOM 2513 C ASP A 475 45.499 71.815 47.794 1.00 16.28 A
ATOM 2514 0 ASP A 475 45.434 72.577 46.829 1.00 17.72 A
ATOM 2515 N ASN A 476 46.573 71.069 48.046 1.00 14.25 A
ATOM 2516 CA ASN A 476 47.732 71.124 47.163 1.00 13.50 A
ATOM 2517 CB ASN A 476 48.905 70.322 47.743 1.00 13.93 A
ATOM 2518 CG ASN A 476 49.477 70.947 49.003 1.00 14.86 A
ATOM 2519 OD1 ASN A 476 49.270 72.127 49.275 1.00 16.38 A
ATOM 2520 ND2 ASN A 476 50.220 70.154 49.773 1.00 13.48 A
ATOM 2521 C ASN A 476 47.395 70.601 45.770 1.00 13.52 A
ATOM 2522 0 ASN A 476 47.861 71.143 44.771 1.00 14.91 A
ATOM 2523 N ARG A 477 46.592 69.543 45.709 1.00 13.19 A
ATOM 2524 CA ARG A 477 46.193 68.971 44.427 1.00 14.37 A
ATOM 2525 CB ARG A 477 45.291 67.757 44.637 1.00 17.12 A
ATOM 2526 CG ARG A 477 44.700 67.224 43.338 1.00 18.32 A
ATOM 2527 CD ARG A 477 43.466 66.371 43.596 1.00 21.12 A
ATOM 2528 NE ARG A 477 42.925 65.816 42.357 1.00 22.13 A
ATOM 2529 CZ ARG A 477 42.224 66.501 41.456 1.00 22.06 A
ATOM 2530 NH1 ARG A 477 41.952 67.787 41.641 1.00 21.09 A
ATOM 2531 NH2 ARG A 477 41.808 65.894 40.354 1.00 22.91 A
ATOM 2532 C ARG A 477 45.423 70.009 43.616 1.00 15.11 A
ATOM 2533 0 ARG A 477 45.706 70.235 42.437 1.00 15.63 A
ATOM 2534 N ASN A 478 44.442 70.630 44.259 1.00 15.73 A

ATOM 2535 CA ASN A 478 43.621 71.631 43.593 1.00 16.96 A
ATOM 2536 CB ASN A 478 42.434 72.021 44.478 1.00 18.15 A
ATOM 2537 CG ASN A 478 41.481 70.863 44.707 1.00 18.54 A
ATOM 2538 OD1 ASN A 478 41.353 69.975 43.862 1.00 20.76 A
ATOM 2539 ND2 ASN A 478 40.797 70.872 45.845 1.00 22.28 A
ATOM 2540 C ASN A 478 44.429 72.861 43.207 1.00 16.90 A
ATOM 2541 0 ASN A 478 44.149 73.500 42.193 1.00 18.68 A
ATOM 2542 N TRP A 479 45.437 73.194 44.006 1.00 17.18 A
ATOM 2543 CA TRP A 479 46.268 74.345 43.694 1.00 17.07 A
ATOM 2544 CB TRP A 479 47.273 74.618 44.815 1.00 17.06 A
ATOM 2545 CG TRP A 479 48.108 75.835 44.549 1.00 18.72 A
ATOM 2546 CD2 TRP A 479 49.377 75.875 43.887 1.00 20.43 A
ATOM 2547 CE2 TRP A 479 49.758 77.230 43.793 1.00 20.82 A
ATOM 2548 CE3 TRP A 479 50.228 74.897 43.360 1.00 20.34 A
ATOM 2549 CD1 TRP A 479 47.781 77.131 44.827 1.00 19.83 A
ATOM 2550 NE1 TRP A 479 48.767 77.977 44.375 1.00 20.62 A
ATOM 2551 CZ2 TRP A 479 50.953 77.631 43.194 1.00 20.76 A
ATOM 2552 CZ3 TRP A 479 51.413 75.298 42.765 1.00 21.63 A
ATOM 2553 CH2 TRP A 479 51.763 76.653 42.688 1.00 20.93 A
ATOM 2554 C TRP A 479 47.022 74.058 42.401 1.00 16.94 A
ATOM 2555 0 TRP A 479 47.030 74.875 41.480 1.00 18.18 A
ATOM 2556 N TYR A 480 47.659 72.892 42.333 1.00 16.87 A
ATOM 2557 CA TYR A 480 48.406 72.523 41.140 1.00 17.45 A
ATOM 2558 CB TYR A 480 49.139 71.194 41.356 1.00 17.97 A
ATOM 2559 CG TYR A 480 50.594 71.365 41.729 1.00 18.22 A
ATOM 2560 CD1 TYR A 480 51.489 71.951 40.845 1.00 19.46 A
ATOM 2561 CE1 TYR A 480 52.825 72.100 41.170 1.00 20.17 A
ATOM 2562 CD2 TYR A 480 51.076 70.933 42.958 1.00 19.67 A
ATOM 2563 CE2 TYR A 480 52.414 71.078 43.291 1.00 20.51 A
ATOM 2564 CZ TYR A 480 53.281 71.660 42.392 1.00 20.12 A
ATOM 2565 OH TYR A 480 54.613 71.789 42.707 1.00 22.48 A
ATOM 2566 C TYR A 480 47.518 72.436 39.906 1.00 17.60 A
ATOM 2567 0 TYR A 480 47.934 72.811 38.809 1.00 19.22 A
ATOM 2568 N GLN A 481 46.295 71.948 40.078 1.00 17.95 A
ATOM 2569 CA GLN A 481 45.387 71.840 38.943 1.00 18.98 A
ATOM 2570 CB GLN A 481 44.106 71.102 39.339 1.00 17.87 A
ATOM 2571 CG GLN A 481 43.209 70.782 38.146 1.00 19.28 A
ATOM 2572 CD GLN A 481 42.073 69.844 38.492 1.00 19.66 A
ATOM 2573 OE1 GLN A 481 41.888 68.810 37.845 1.00 21.59 A
ATOM 2574 NE2 GLN A 481 41.303 70.198 39.511 1.00 18.91 A
ATOM 2575 C GLN A 481 45.045 73.230 38.418 1.00 19.80 A
ATOM 2576 0 GLN A 481 44.938 73.434 37.209 1.00 21.26 A
ATOM 2577 N SER A 482 44.893 74.189 39.325 1.00 20.93 A
ATOM 2578 CA SER A 482 44.557 75.557 38.932 1.00 22.05 A
ATOM 2579 CB SER A 482 44.059 76.357 40.139 1.00 20.51 A
ATOM 2580 OG SER A 482 45.125 76.687 41.015 1.00 23.08 A
ATOM 2581 C SER A 482 45.741 76.286 38.306 1.00 22.53 A
ATOM 2582 0 SER A 482 45.570 77.337 37.683 1.00 24.61 A
ATOM 2583 N MET A 483 46.938 75.730 38.465 1.00 22.77 A
ATOM 2584 CA MET A 483 48.143 76.345 37.915 1.00 22.53 A
ATOM 2585 CB MET A 483 49.306 76.203 38.898 1.00 21.83 A
ATOM 2586 CG MET A 483 49.141 76.998 40.182 1.00 24.67 A
ATOM 2587 SD MET A 483 49.123 78.781 39.907 1.00 24.46 A
ATOM 2588 CE MET A 483 50.853 79.097 39.558 1.00 27.42 A
ATOM 2589 C MET A 483 48.545 75.756 36.570 1.00 23.15 A
ATOM 2590 0 MET A 483 49.557 76.155 35.989 1.00 24.17 A
ATOM 2591 N ILE A 484 47.767 74.798 36.080 1.00 22.82 A
ATOM 2592 CA ILE A 484 48.064 74.194 34.787 1.00 24.95 A
ATOM 2593 CB ILE A 484 47.072 73.058 34.444 1.00 23.62 A
ATOM 2594 CG2 ILE A 484 47.307 72.572 33.026 1.00 24.43 A
ATOM 2595 CG1 ILE A 484 47.238 71.899 35.428 1.00 22.63 A
ATOM 2596 CD1 ILE A 484 46.251 70.775 35.221 1.00 20.73 A
ATOM 2597 C ILE A 484 47.943 75.295 33.739 1.00 25.95 A
ATOM 2598 0 ILE A 484 46.938 76.000 33.682 1.00 25.99 A
ATOM 2599 N PRO A 485 48.982 75.468 32.909 1.00 25.89 A
ATOM 2600 CD PRO A 485 50.237 74.695 32.922 1.00 26.30 A
ATOM 2601 CA PRO A 485 49.007 76.487 31.855 1.00 26.49 A

ATOM 2602 CB PRO A 485 50.229 76.092 31.035 1.00 26.74 A
ATOM 2603 CG PRO A 485 51.158 75.560 32.087 1.00 26.49 A
ATOM 2604 C PRO A 485 47.729 76.519 31.019 1.00 27.33 A
ATOM 2605 0 PRO A 485 47.446 75.591 30.257 1.00 28.98 A
ATOM 2606 N GLN A 500 57.823 61.166 25.823 1.00 27.23 A
ATOM 2607 CA GLN A 500 56.906 60.889 26.925 1.00 27.11 A
ATOM 2608 CB GLN A 500 57.203 59.535 27.562 1.00 26.34 A
ATOM 2609 CG GLN A 500 56.914 58.342 26.657 1.00 26.66 A
ATOM 2610 CD GLN A 500 57.025 57.026 27.409 1.00 28.35 A
ATOM 2611 OE1 GLN A 500 57.979 56.804 28.133 1.00 28.45 A
ATOM 2612 NE2 GLN A 500 56.166 56.009 27.357 1.00 29.09 A
ATOM 2613 C GLN A 500 56.949 61.974 27.995 1.00 27.94 A
ATOM 2614 0 GLN A 500 58.022 62.391 28.427 1.00 28.09 A
ATOM 2615 N GLY A 501 55.770 62.423 28.419 1.00 27.36 A
ATOM 2616 CA GLY A 501 55.688 63.453 29.439 1.00 27.32 A
ATOM 2617 C GLY A 501 56.098 62.919 30.799 1.00 27.53 A
ATOM 2618 0 GLY A 501 56.269 61.721 30.950 1.00 27.48 A
ATOM 2619 N LEU A 502 56.218 63.771 31.790 1.00 25.91 A
ATOM 2620 CA LEU A 502 56.512 63.326 33.137 1.00 25.00 A
ATOM 2621 C LEU A 502 55.686 62.081 33.495 1.00 25.16 A
ATOM 2622 0 LEU A 502 56.218 60.995 33.676 1.00 25.33 A
ATOM 2623 CB LEU A 502 56.189 64.430 34.157 1.00 24.37 A
ATOM 2624 CG LEU A 502 57.152 65.640 34.297 1.00 20.00 A
ATOM 2625 CD1 LEU A 502 56.648 66.625 35.340 1.00 20.00 A
ATOM 2626 CD2 LEU A 502 58.549 65.154 34.650 1.00 20.00 A
ATOM 2627 N MET A 503 54.414 62.336 33.603 1.00 24.22 A
ATOM 2628 CA MET A 503 53.511 61.306 33.995 1.00 24.40 A
ATOM 2629 CB MET A 503 52.078 61.835 34.086 1.00 26.37 A
ATOM 2630 CG MET A 503 51.082 60.804 34.575 1.00 29.12 A
ATOM 2631 SD MET A 503 51.581 60.069 36.144 1.00 29.58 A
ATOM 2632 CE MET A 503 51.097 61.354 37.299 1.00 31.34 A
ATOM 2633 C MET A 503 53.554 60.104 33.057 1.00 24.49 A
ATOM 2634 0 MET A 503 53.471 58.960 33.502 1.00 24.24 A
ATOM 2635 N GLU A 504 53.680 60.363 31.759 1.00 24.16 A
ATOM 2636 CA GLU A 504 53.740 59.279 30.783 1.00 24.81 A
ATOM 2637 C GLU A 504 54.938 58.385 31.090 1.00 25.80 A
ATOM 2638 0 GLU A 504 54.813 57.160 31.152 1.00 26.68 A
ATOM 2639 CB GLU A 504 53.788 59.840 29.362 1.00 24.38 A
ATOM 2640 CG GLU A 504 52.586 60.691 28.990 1.00 20.00 A
ATOM 2641 CD GLU A 504 52.669 61.230 27.575 1.00 20.00 A
ATOM 2642 OE1 GLU A 504 53.667 60.937 26.886 1.00 20.00 A
ATOM 2643 OE2 GLU A 504 51.726 61.928 27.150 1.00 20.00 A
ATOM 2644 N LYS A 505 56.030 58.881 31.278 1.00 25.07 A
ATOM 2645 CA LYS A 505 57.326 58.286 31.598 1.00 24.86 A
ATOM 2646 C LYS A 505 57.176 57.297 32.750 1.00 24.44 A
ATOM 2647 0 LYS A 505 57.437 56.100 32.603 1.00 24.95 A
ATOM 2648 CB LYS A 505 58.352 59.375 31.913 1.00 23.71 A
ATOM 2649 CG LYS A 505 58.594 60.348 30.772 1.00 20.00 A
ATOM 2650 CD LYS A 505 59.787 61.245 31.055 1.00 20.00 A
ATOM 2651 CE LYS A 505 60.028 62.218 29.914 1.00 20.00 A
ATOM 2652 NZ LYS A 505 61.195 63.104 30.177 1.00 20.00 A
ATOM 2653 N PHE A 506 56.589 58.121 33.897 1.00 23.30 A
ATOM 2654 CA PHE A 506 56.349 57.467 35.176 1.00 23.05 A
ATOM 2655 CB PHE A 506 55.595 58.411 36.113 1.00 22.73 A
ATOM 2656 CG PHE A 506 55.268 57.805 37.447 1.00 22.31 A
ATOM 2657 CD1 PHE A 506 56.253 57.627 38.404 1.00 22.89 A
ATOM 2658 CD2 PHE A 506 53.977 57.394 37.735 1.00 22.89 A
ATOM 2659 CE1 PHE A 506 55.957 57.048 39.625 1.00 23.23 A
ATOM 2660 CE2 PHE A 506 53.674 56.815 38.951 1.00 22.14 A
ATOM 2661 CZ PHE A 506 54.666 56.642 39.896 1.00 22.20 A
ATOM 2662 C PHE A 506 55.527 56.196 34.970 1.00 24.17 A
ATOM 2663 0 PHE A 506 55.915 55.114 35.415 1.00 24.62 A
ATOM 2664 N GLN A 507 54.394 56.336 34.288 1.00 24.03 A
ATOM 2665 CA GLN A 507 53.512 55.205 34.023 1.00 25.65 A
ATOM 2666 C GLN A 507 54.185 54.150 33.149 1.00 26.46 A
ATOM 2667 0 GLN A 507 54.005 52.951 33.364 1.00 28.00 A
ATOM 2668 CB GLN A 507 52.215 55.680 33.366 1.00 26.09 A

ATOM 2669 CG GLN A 507 51.260 56.383 34.315 1.00 20.00 A
ATOM 2670 CD GLN A 507 50.230 55.442 34.907 1.00 20.00 A
ATOM 2671 OE1 GLN A 507 50.222 54.248 34.610 1.00 20.00 A
ATOM 2672 NE2 GLN A 507 49.269 55.764 35.764 1.00 20.00 A
ATOM 2673 N PHE A 508 54.962 54.601 32.168 1.00 26.64 A
ATOM 2674 CA PHE A 508 55.662 53.700 31.257 1.00 26.96 A
ATOM 2675 C PHE A 508 56.637 52.780 31.990 1.00 26.74 A
ATOM 2676 0 PHE A 508 56.581 51.559 31.836 1.00 27.49 A
ATOM 2677 CB PHE A 508 56.426 54.496 30.198 1.00 25.48 A
ATOM 2678 CG PHE A 508 56.589 53.770 28.895 1.00 20.00 A
ATOM 2679 CD1 PHE A 508 55.486 53.433 28.131 1.00 20.00 A
ATOM 2680 CD2 PHE A 508 57.859 53.423 28.434 1.00 20.00 A
ATOM 2681 CE1 PHE A 508 55.636 52.765 26.932 1.00 20.00 A
ATOM 2682 CE2 PHE A 508 58.009 52.755 27.235 1.00 20.00 A
ATOM 2683 CZ PHE A 508 56.898 52.426 26.484 1.00 20.00 A
ATOM 2684 N GLU A 509 57.375 53.138 32.892 1.00 26.57 A
ATOM 2685 CA GLU A 509 58.236 52.244 33.658 1.00 26.37 A
ATOM 2686 CB GLU A 509 59.300 53.031 34.426 1.00 25.10 A
ATOM 2687 CG GLU A 509 60.275 53.824 33.575 1.00 26.10 A
ATOM 2688 CD GLU A 509 61.477 54.282 34.382 1.00 25.19 A
ATOM 2689 OE1 GLU A 509 61.303 54.568 35.584 1.00 29.75 A
ATOM 2690 OE2 GLU A 509 62.588 54.366 33.818 1.00 27.78 A
ATOM 2691 C GLU A 509 57.449 51.401 34.651 1.00 28.55 A
ATOM 2692 0 GLU A 509 57.540 50.173 34.643 1.00 29.93 A
ATOM 2693 N THR A 510 56.686 52.064 35.515 1.00 28.05 A
ATOM 2694 CA THR A 510 55.895 51.362 36.521 1.00 28.54 A
ATOM 2695 CB THR A 510 55.042 52.346 37.356 1.00 27.27 A
ATOM 2696 OG1 THR A 510 54.443 51.649 38.457 1.00 29.38 A
ATOM 2697 CG2 THR A 510 53.952 52.964 36.504 1.00 26.53 A
ATOM 2698 C THR A 510 54.982 50.322 35.872 1.00 29.49 A
ATOM 2699 0 THR A 510 54.894 49.202 36.416 1.00 29.59 A
ATOM 2700 OXT THR A 510 54.364 50.638 34.832 1.00 28.89 A
ATOM 2701 CB ARG B 155 85.015 19.003 21.577 1.00 24.19 B
ATOM 2702 CG ARG B 155 84.934 20.238 22.448 1.00 23.50 B
ATOM 2703 CD ARG B 155 84.822 21.492 21.606 1,00 21.83 B
ATOM 2704 NE ARG B 155 84.724 22.687 22.435 1.00 19.82 B
ATOM 2705 CZ ARG B 155 84.571 23.918 21.957 1.00 19.67 B
ATOM 2706 NH1 ARG B 155 84.498 24.120 20.645 1.00 21.34 B
ATOM 2707 NH2 ARG B 155 84.491 24.945 22.791 1.00 20.32 B
ATOM 2708 C ARG B 155 84.244 17.539 23.451 1.00 24.29 B
ATOM 2709 0 ARG B 155 84.400 18.070 24.551 1.00 25.28 B
ATOM 2710 N ARG B 155 85.300 16.557 21.415 1.00 26.08 B
ATOM 2711 CA ARG B 155 85.285 17.714 22.350 1.00 24.35 B
ATOM 2712 N PHE B 156 83.188 16.786 23.155 1.00 22.89 B
ATOM 2713 CA PHE B 156 82.122 16.538 24.124 1.00 21.98 B
ATOM 2714 CB PHE B 156 80.862 16.043 23.410 1.00 19.58 B
ATOM 2715 CG PHE B 156 80.162 17.100 22.613 1.00 19.92 B
ATOM 2716 CD1 PHE B 156 79.307 18.002 23.233 1.00 20.93 B
ATOM 2717 CD2 PHE B 156 80.366 17.206 21.249 1.00 22.26 B
ATOM 2718 CE1 PHE B 156 78.671 18.988 22.505 1.00 21.48 B
ATOM 2719 CE2 PHE B 156 79.731 18.193 20.512 1.00 23.50 B
ATOM 2720 CZ PHE B 156 78.883 19.084 21.142 1.00 22.42 B
ATOM 2721 C PHE B 156 82.550 15.509 25.159 1.00 22.86 B
ATOM 2722 0 PHE B 156 81.908 15.357 26.202 1.00 24.31 B
ATOM 2723 N GLY B 157 83.638 14.804 24.868 1.00 23.36 B
ATOM 2724 CA GLY B 157 84.126 13.796 25.790 1.00 23.56 B
ATOM 2725 C GLY B 157 83.679 12.396 25.411 1.00 24.97 B
ATOM 2726 0 GLY B 157 83.722 11.477 26.232 1.00 26.71 B
ATOM 2727 N VAL B 158 83.248 12.229 24.164 1.00 24.24 B
ATOM 2728 CA VAL B 158 82.795 10.927 23.688 1.00 23.64 B
ATOM 2729 CB VAL B 158 81.580 11.067 22.741 1.00 22.81 B
ATOM 2730 CG1 VAL B 158 81.103 9.692 22.295 1.00 22.53 B
ATOM 2731 CG2 VAL B 158 80.457 11.813 23.438 1.00 23.22 B
ATOM 2732 C VAL B 158 83.907 10.189 22.948 1.00 24.16 B
ATOM 2733 0 VAL B 158 84.453 10.686 21.961 1.00 25.45 B
ATOM 2734 N ASN B 159 84.245 9.001 23.438 1.00 23.84 B
ATOM 2735 CA ASN B 159 85.277 8.184 22.812 1.00 24.89 B

ATOM 2736 CB ASN B 159 85.534 6.925 23.645 1.00 25.27 B
ATOM 2737 CG ASN B 159 86.188 7.230 24.984 1.00 27.41 B
ATOM 2738 OD1 ASN B 159 86.201 6.391 25.887 1.00 31.29 B
ATOM 2739 ND2 ASN B 159 86.746 8.429 25.113 1.00 29.14 B
ATOM 2740 C ASN B 159 84.789 7.797 21.420 1.00 24.42 B
ATOM 2741 0 ASN B 159 83.622 7.449 21.238 1.00 24.56 B
ATOM 2742 N THR B 160 85.681 7.867 20.438 1.00 25.41 B
ATOM 2743 CA THR B 160 85.327 7.528 19.065 1.00 25.78 B
ATOM 2744 CB THR B 160 86.583 7.452 18.177 1.00 26.84 B
ATOM 2745 OG1 THR B 160 87.179 8.752 18.086 1.00 30.04 B
ATOM 2746 CG2 THR B 160 86.224 6.965 16.784 1.00 27.12 B
ATOM 2747 C THR B 160 84.571 6.206 18.964 1.00 25.75 B
ATOM 2748 0 THR B 160 83.667 6.056 18.141 1.00 26.05 B
ATOM 2749 N GLU B 161 84.940 5.254 19.814 1.00 24.76 B
ATOM 2750 CA GLU B 161 84.313 3.938 19.817 1.00 25.15 B
ATOM 2751 C GLU B 161 82.869 3.935 20.310 1.00 25.80 B
ATOM 2752 0 GLU B 161 82.123 2.994 20.035 1.00 25.86 B
ATOM 2753 CB GLU B 161 85.098 2.973 20.706 1.00 24.94 B
ATOM 2754 CG GLU B 161 86.539 2.764 20.274 1.00 20.00 B
ATOM 2755 CD GLU B 161 87.285 1.803 21.178 1.00 20.00 B
ATOM 2756 OE1 GLU B 161 86.674 1.300 22.143 1.00 20.00 B
ATOM 2757 OE2 GLU B 161 88.480 1.553 20.921 1.00 20.00 B
ATOM 2758 N ASN B 162 82.469 4.977 21.034 1.00 24.54 B
ATOM 2759 CA ASN B 162 81.106 5.050 21.558 1.00 23.42 B
ATOM 2760 CB ASN B 162 81.113 5.526 23.013 1.00 24.01 B
ATOM 2761 CG ASN B 162 81.716 4.510 23.959 1.00 24.31 B
ATOM 2762 OD1 ASN B 162 81.408 3.321 23.888 1.00 28.88 B
ATOM 2763 ND2 ASN B 162 82.569 4.977 24.864 1.00 25.80 B
ATOM 2764 C ASN B 162 80.167 5.943 20.752 1.00 23.07 B
ATOM 2765 0 ASN B 162 79.006 6.115 21.123 1.00 23.17 B
ATOM 2766 N GLU B 163 80.656 6.502 19.651 1.00 22.51 B
ATOM 2767 CA GLU B 163 79.832 7.371 18.817 1.00 24.08 B
ATOM 2768 C GLU B 163 78.535 6.686 18.391 1.00 25.06 B
ATOM 2769 0 GLU B 163 77.478 7.320 18.335 1.00 25.48 B
ATOM 2770 CB GLU B 163 80.613 7.828 17.585 1.00 23.35 B
ATOM 2771 CG GLU B 163 81.746 8.793 17.892 1.00 20.00 B
ATOM 2772 CD GLU B 163 81.255 10.201 18.162 1.00 20.00 B
ATOM 2773 OE1 GLU B 163 80.028 10.422 18.110 1.00 20.00 B
ATOM 2774 OE2 GLU B 163 82.098 11.083 18.427 1.00 20.00 B
ATOM 2775 N ASP B 164 78.618 5.391 18.097 1.00 24.90 B
ATOM 2776 CA ASP B 164 77.451 4.621 17.673 1.00 26.01 B
ATOM 2777 CB ASP B 164 77.893 3.295 17.049 1.00 27.20 B
ATOM 2778 CG ASP B 164 78.609 3.488 15.728 1.00 29.72 B
ATOM 2779 OD1 ASP B 164 78.016 4.110 14.819 1.00 32.07 B
ATOM 2780 OD2 ASP B 164 79.761 3.022 15.596 1.00 32.92 B
ATOM 2781 C ASP B 164 76.457 4.351 18.799 1.00 25.49 B
ATOM 2782 0 ASP B 164 75.246 4.438 18.597 1.00 27.12 B
ATOM 2783 N HIS B 165 76.965 4.010 19.979 1.00 24.39 B
ATOM 2784 CA HIS B 165 76.098 3.743 21.120 1.00 24.02 B
ATOM 2785 CB HIS B 165 76.912 3.225 22.308 1.00 24.80 B
ATOM 2786 CG HIS B 165 77.365 1.806 22.158 1.00 26.39 B
ATOM 2787 CD2 HIS B 165 77.176 0.911 21.161 1.00 26.51 B
ATOM 2788 ND1 HIS B 165 78.095 1.152 23.128 1.00 26.98 B
ATOM 2789 CE1 HIS B 165 78.333 -0.086 22.735 1.00 25.33 B
ATOM 2790 NE2 HIS B 165 77.786 -0.259 21.545 1.00 26.97 B
ATOM 2791 C HIS B 165 75.376 5.020 21.527 1.00 23.78 B
ATOM 2792 0 HIS B 165 74.185 5.008 21.842 1.00 23.93 B
ATOM 2793 N LEU B 166 76.108 6.126 21.521 1.00 23.16 B
ATOM 2794 CA LEU B 166 75.531 7.408 21.895 1.00 22.75 B
ATOM 2795 CB LEU B 166 76.631 8.470 21.976 1.00 22.64 B
ATOM 2796 CG LEU B 166 76.197 9.870 22.418 1.00 21.89 B
ATOM 2797 CD1 LEU B 166 75.537 10.584 21.261 1.00 25.50 B
ATOM 2798 CD2 LEU B 166 75.257 9.764 23.612 1.00 22.26 B
ATOM 2799 C LEU B 166 74.453 7.825 20.900 1.00 22.78 B
ATOM 2800 0 LEU B 166 73.352 8.203 21.295 1.00 21.99 B
ATOM 2801 N ALA B 167 74.766 7.751 19.610 1.00 22.22 B
ATOM 2802 CA ALA B 167 73.802 8.128 18.581 1.00 22.86 B

ATOM 2803 CB ALA B 167 74.422 7.972 17.198 1.00 21.86 B
ATOM 2804 C ALA B 167 72.548 7.267 18.699 1.00 23.08 B
ATOM 2805 0 ALA B 167 71.435 7.728 18.450 1.00 24.20 B
ATOM 2806 N LYS B 168 72.743 6.012 19.087 1.00 23.38 B
ATOM 2807 CA LYS B 168 71.649 5.065 19.258 1.00 24.21 B
ATOM 2808 CB LYS B 168 72.215 3.706 19.676 1.00 24.39 B
ATOM 2809 CG LYS B 168 71.192 2.712 20.194 1.00 24.92 B
ATOM 2810 CD LYS B 168 71.832 1.839 21.269 1.00 27.56 B
ATOM 2811 CE LYS B 168 72.373 2.709 22.404 1.00 25.91 B
ATOM 2812 NZ LYS B 168 73.151 1.960 23.423 1.00 29.53 B
ATOM 2813 C LYS B 168 70.681 5.575 20.321 1.00 23.73 B
ATOM 2814 0 LYS B 168 69.471 5.630 20.098 1.00 25.71 B
ATOM 2815 N GLU B 169 71.223 5.949 21.476 1.00 23.38 B
ATOM 2816 CA GLU B 169 70.408 6.463 22.571 1.00 22.17 B
ATOM 2817 CB GLU B 169 71.275 6.741 23.800 1.00 22.63 B
ATOM 2818 CG GLU B 169 71.129 5.730 24.928 1.00 26.70 B
ATOM 2819 CD GLU B 169 69.678 5.436 25.276 1.00 26.56 B
ATOM 2820 OE1 GLU B 169 68.859 6.381 25.325 1.00 30.37 B
ATOM 2821 OE2 GLU B 169 69.356 4.252 25.509 1.00 29.65 B
ATOM 2822 C GLU B 169 69.693 7.748 22.170 1.00 20.80 B
ATOM 2823 0 GLU B 169 68.511 7.929 22.461 1.00 21.00 B
ATOM 2824 N LEU B 170 70.414 8.642 21.502 1.00 21.11 B
ATOM 2825 CA LEU B 170 69.840 9.911 21.081 1.00 21.11 B
ATOM 2826 CB LEU B 170 70.929 10.822 20.514 1.00 20.36 B
ATOM 2827 CG LEU B 170 71.924 11.313 21.567 1.00 20.03 B
ATOM 2828 CD1 LEU B 170 72.894 12.289 20.933 1.00 19.84 B
ATOM 2829 CD2 LEU B 170 71.168 11.978 22.712 1.00 20.21 B
ATOM 2830 C LEU B 170 68.705 9.761 20.078 1.00 21.80 B
ATOM 2831 0 LEU B 170 68.011 10.731 19.767 1.00 22.98 B
ATOM 2832 N GLU B 171 68.510 8.551 19.566 1.00 22.07 B
ATOM 2833 CA GLU B 171 67.422 8.323 18.625 1.00 22.54 B
ATOM 2834 CB GLU B 171 67.398 6.866 18.157 1.00 23.58 B
ATOM 2835 CG GLU B 171 68.431 6.527 17.104 1.00 25.26 B
ATOM 2836 CD GLU B 171 68.258 5.118 16.564 1.00 28.30 B
ATOM 2837 OE1 GLU B 171 67.140 4.790 16.112 1.00 30.60 B
ATOM 2838 OE2 GLU B 171 69.238 4.341 16.590 1.00 31,63 B
ATOM 2839 C GLU B 171 66.109 8.643 19.326 1.00 21.58 B
ATOM 2840 0 GLU B 171 65.120 8.996 18.686 1.00 22.06 B
ATOM 2841 N ASP B 172 66.114 8.523 20.650 1.00 21.60 B
ATOM 2842 CA ASP B 172 64.924 8.787 21.450 1.00 20.55 B
ATOM 2843 CB ASP B 172 64.809 7.756 22.575 1.00 23.36 B
ATOM 2844 CG ASP B 172 64.723 6.333 22.057 1.00 27.58 B
ATOM 2845 OD1 ASP B 172 63.862 6.062 21.193 1.00 30.94 B
ATOM 2846 OD2 ASP B 172 65.514 5.484 22.521 1.00 30.56 B
ATOM 2847 C ASP B 172 64.901 10.192 22.049 1.00 19.21 B
ATOM 2848 0 ASP B 172 64.195 10.443 23.025 1.00 20.00 B
ATOM 2849 N LEU B 173 65.664 11.103 21.453 1,00 18.97 B
ATOM 2850 CA LEU B 173 65.733 12.481 21.927 1.00 17.67 B
ATOM 2851 CB LEU B 173 66.581 13.322 20.974 1.00 18.94 B
ATOM 2852 CG LEU B 173 66.725 14.793 21.363 1.00 19.48 B
ATOM 2853 CD1 LEU B 173 67.480 14.884 22.676 1.00 20.01 B
ATOM 2854 CD2 LEU B 173 67.453 15.554 20.267 1.00 21.37 B
ATOM 2855 C LEU B 173 64.364 13.143 22.076 1.00 17.35 B
ATOM 2856 0 LEU B 173 64.129 13.887 23.024 1.00 17.56 B
ATOM 2857 N ASN B 174 63.461 12.874 21.140 1.00 17.52 B
ATOM 2858 CA ASN B 174 62.140 13.491 21.185 1.00 18.76 B
ATOM 2859 CB ASN B 174 61.650 13.783 19,760 1.00 18.55 B
ATOM 2860 CG ASN B 174 62.678 14.535 18.928 1.00 19.92 B
ATOM 2861 OD1 ASN B 174 63.334 15.457 19.411 1.00 21.88 B
ATOM 2862 ND2 ASN B 174 62.813 14.148 17.665 1.00 24.47 B
ATOM 2863 C ASN B 174 61.099 12.657 21.922 1.00 18.50 B
ATOM 2864 0 ASN B 174 59.901 12.928 21.830 1.00 19.21 B
ATOM 2865 N LYS B 175 61.557 11.661 22.673 1.00 19.03 B
ATOM 2866 CA LYS B 175 60.646 10.791 23.404 1.00 19.39 B
ATOM 2867 CB LYS B 175 60.831 9.341 22.960 1.00 21.10 B
ATOM 2868 CG LYS B 175 60.706 9.132 21.466 1.00 22.98 B
ATOM 2869 CD LYS B 175 60.873 7.669 21.121 1.00 24.14 B

ATOM 2870 CE LYS B 175 60.807 7.439 19.623 1.00 25.74 B
ATOM 2871 NZ LYS B 175 60.977 5.994 19.308 1.00 28.38 B
ATOM 2872 C LYS B 175 60.824 10.872 24.910 1.00 17.95 B
ATOM 2873 0 LYS B 175 61.913 11.147 25.413 1.00 19.34 B
ATOM 2874 N TRP B 176 59.735 10.613 25.622 1.00 18.25 B
ATOM 2875 CA TRP B 176 59.715 10.639 27.077 1.00 17.79 B
ATOM 2876 CB TRP B 176 58.267 10.496 27.553 1.00 16.96 B
ATOM 2877 CG TRP B 176 57.983 11.021 28.929 1.00 16.64 B
ATOM 2878 CD2 TRP B 176 58.033 12.390 29.351 1.00 16.66 B
ATOM 2879 CE2 TRP B 176 57.610 12.427 30.696 1.00 16.99 B
ATOM 2880 CE3 TRP B 176 58.392 13.588 28.721 1.00 17.54 B
ATOM 2881 CD1 TRP B 176 57.553 10.305 30.007 1.00 17.09 B
ATOM 2882 NE1 TRP B 176 57.324 11.141 31.072 1.00 17.25 B
ATOM 2883 CZ2 TRP B 176 57.535 13.613 31.422 1.00 15.39 B
ATOM 2884 CZ3 TRP B 176 58.317 14.766 29.443 1.00 17.16 B
ATOM 2885 CH2 TRP B 176 57.891 14.770 30.780 1.00 14,84 B
ATOM 2886 C TRP B 176 60.567 9.496 27.625 1.00 18.34 B
ATOM 2887 0 TRP B 176 61.042 9.550 28.757 1.00 20.26 B
ATOM 2888 N GLY B 177 60.778 8.474 26.801 1.00 20.21 B
ATOM 2889 CA GLY B 177 61.549 7.323 27.233 1.00 20.19 B
ATOM 2890 C GLY B 177 63.059 7.386 27.109 1.00 20.06 B
ATOM 2891 0 GLY B 177 63.735 6.392 27.362 1.00 22.63 B
ATOM 2892 N LEU B 178 63.603 8.530 26.710 1.00 19.92 B
ATOM 2893 CA LEU B 178 65.051 8.643 26.591 1.00 18.61 B
ATOM 2894 CB LEU B 178 65.445 10.059 26.161 1.00 18.69 B
ATOM 2895 CG LEU B 178 66.950 10.318 26.027 1.00 20.59 B
ATOM 2896 CD1 LEU B 178 67.224 11.134 24.785 1.00 21.10 B
ATOM 2897 CD2 LEU B 178 67.465 11.032 27.266 1.00 18.71 B
ATOM 2898 C LEU B 178 65.695 8.311 27.932 1.00 18.99 B
ATOM 2899 0 LEU B 178 65.155 8.647 28.986 1.00 20.22 B
ATOM 2900 N ASN B 179 66.839 7.633 27.892 1.00 19.59 B
ATOM 2901 CA ASN B 179 67.554 7.268 29.112 1.00 19.00 B
ATOM 2902 CB ASN B 179 68.039 5.818 29.042 1.00 19.91 B
ATOM 2903 CG ASN B 179 68.558 5.312 30.378 1.00 20.81 B
ATOM 2904 ODl ASN B 179 69.379 5.960 31.022 1.00 20.85 B
ATOM 2905 ND2 ASN B 179 68.079 4.147 30.799 1.00 22.81 B
ATOM 2906 C ASN B 179 68.749 8.200 29.243 1.00 18.92 B
ATOM 2907 0 ASN B 179 69.782 7.994 28.605 1.00 19.10 B
ATOM 2908 N ILE B 180 68.610 9.219 30.082 1.00 18.12 B
ATOM 2909 CA ILE B 180 69.678 10.193 30.260 1.00 16.99 B
ATOM 2910 CB ILE B 180 69.154 11.453 31.008 1.00 17.99 B
ATOM 2911 CG2 ILE B 180 68.982 11.156 32.483 1.00 17.57 B
ATOM 2912 CG1 ILE B 180 70.119 12.623 30.801 1.00 17.99 B
ATOM 2913 CD1 ILE B 180 70.199 13.104 29.350 1.00 20.60 B
ATOM 2914 C ILE B 180 70.888 9.603 30.989 1.00 15.91 B
ATOM 2915 0 ILE B 180 72.007 10.095 30.845 1.00 17.37 B
ATOM 2916 N PHE B 181 70.668 8.544 31.762 1.00 17.29 B
ATOM 2917 CA PHE B 181 71.760 7.901 32.488 1.00 17.59 B
ATOM 2918 CB PHE B 181 71.219 6.787 33.383 1.00 17.20 B
ATOM 2919 CG PHE B 181 70.374 7.286 34.513 1.00 18.19 B
ATOM 2920 CD1 PHE B 181 70.959 7.796 35.660 1.00 19.64 B
ATOM 2921 CD2 PHE B 181 68.995 7.287 34.411 1.00 20.43 B
ATOM 2922 CE1 PHE B 181 70.183 8.301 36.682 1.00 20.68 B
ATOM 2923 CE2 PHE B 181 68.212 7.790 35.430 1.00 20.48 B
ATOM 2924 CZ PHE B 181 68.809 8.299 36.567 1.00 19.24 B
ATOM 2925 C PHE B 181 72.763 7.326 31.498 1.00 18.24 B
ATOM 2926 0 PHE B 181 73.970 7.374 31.729 1.00 20.46 B
ATOM 2927 N ASN B 182 72.260 6.784 30.390 1.00 19.24 B
ATOM 2928 CA ASN B 182 73.141 6.219 29.376 1.00 19.37 B
ATOM 2929 CB ASN B 182 72.349 5.398 28.358 1.00 19.84 B
ATOM 2930 CG ASN B 182 71.730 4.158 28.970 1.00 22.83 B
ATOM 2931 OD1 ASN B 182 72.260 3.596 29.930 1.00 24.83 B
ATOM 2932 ND2 ASN B 182 70.610 3.716 28.408 1.00 24.12 B
ATOM 2933 C ASN B 182 73.904 7.322 28.661 1.00 19.41 B
ATOM 2934 0 ASN B 182 75.088 7.180 28.380 1.00 19.81 B
ATOM 2935 N VAL B 183 73.219 8.424 28.366 1.00 19.07 B
ATOM 2936 CA VAL B 183 73.864 9.547 27.699 1.00 18.04 B

ATOM 2937 CB VAL B 183 72.884 10.730 27.517 1.00 18.09 B
ATOM 2938 CG1 VAL B 183 73.593 11.888 26.820 1.00 20.77 B
ATOM 2939 CG2 VAL B 183 71.664 10.282 26.712 1.00 17.63 B
ATOM 2940 C VAL B 183 75.064 10.019 28.518 1.00 17.93 B
ATOM 2941 0 VAL B 183 76.127 10.315 27.972 1.00 19.33 B
ATOM 2942 N ALA B 184 74.894 10.084 29.835 1.00 17.20 B
ATOM 2943 CA ALA B 184 75.966 10.521 30.723 1.00 18.84 B
ATOM 2944 CB ALA B 184 75.456 10.602 32.159 1.00 20.06 B
ATOM 2945 C ALA B 184 77.150 9.563 30.645 1.00 19.77 B
ATOM 2946 0 ALA B 184 78.303 9.988 30.590 1.00 19.55 B
ATOM 2947 N GLY B 185 76.857 8.268 30.631 1.00 19.84 B
ATOM 2948 CA GLY B 185 77.917 7.279 30.560 1.00 20.49 B
ATOM 2949 C GLY B 185 78.765 7.358 29.303 1.00 21.38 B
ATOM 2950 0 GLY B 185 79.970 7.119 29.350 1.00 23.91 B
ATOM 2951 N TYR B 186 78.146 7.692 28.176 1.00 21.11 B
ATOM 2952 CA TYR B 186 78.873 7.785 26.915 1.00 21.18 B
ATOM 2953 CB TYR B 186 77.938 7.508 25.734 1.00 21.47 B
ATOM 2954 CG TYR B 186 77.361 6.113 25.709 1.00 22.97 B
ATOM 2955 CD1 TYR B 186 78.181 4.999 25.830 1.00 24.76 B
ATOM 2956 CE1 TYR B 186 77.658 3.716 25.782 1.00 25.32 B
ATOM 2957 CD2 TYR B 186 75.997 5.909 25.539 1.00 24.05 B
ATOM 2958 CE2 TYR B 186 75.465 4.633 25.490 1.00 24.48 B
ATOM 2959 CZ TYR B 186 76.300 3.541 25.612 1.00 26.33 B
ATOM 2960 OH TYR B 186 75.774 2.270 25.561 1.00 28.76 B
ATOM 2961 C TYR B 186 79.552 9.127 26.686 1.00 21.96 B
ATOM 2962 0 TYR B 186 80.395 9.249 25.801 1.00 20.82 B
ATOM 2963 N SER B 187 79.198 10.132 27.481 1.00 20.97 B
ATOM 2964 CA SER B 187 79.772 11.464 27.303 1.00 22.17 B
ATOM 2965 CB SER B 187 78.647 12.491 27.171 1.00 21.61 B
ATOM 2966 OG SER B 187 77.867 12.524 28.354 1.00 21.70 B
ATOM 2967 C SER B 187 80.741 11.933 28.387 1.00 22.18 B
ATOM 2968 0 SER B 187 80.903 13.137 28.587 1.00 21.91 B
ATOM 2969 N HIS B 188 81.385 11.002 29.082 1.00 22.65 B
ATOM 2970 CA HIS B 188 82.332 11.380 30.129 1.00 22.66 B
ATOM 2971 CB HIS B 188 83.542 12.087 29.501 1.00 24.04 B
ATOM 2972 CG HIS B 188 84.709 12.240 30.427 1.00 26.08 B
ATOM 2973 CD2 HIS B 188 85.953 11.706 30.387 1.00 26.32 B
ATOM 2974 ND1 HIS B 188 84.665 13.019 31.564 1.00 27.31 B
ATOM 2975 CE1 HIS B 188 85.830 12.957 32.184 1.00 27.44 B
ATOM 2976 NE2 HIS B 188 86.629 12.167 31.490 1.00 27.75 B
ATOM 2977 C HIS B 188 81.644 12.306 31.138 1.00 22.22 B
ATOM 2978 0 HIS B 188 82.181 13.344 31.521 1.00 23.24 B
ATOM 2979 N ASN B 189 80.443 11.919 31.552 1.00 21.13 B
ATOM 2980 CA ASN B 189 79.657 12.678 32.517 1.00 20.73 B
ATOM 2981 CB ASN B 189 80.296 12.602 33.903 1.00 22.75 B
ATOM 2982 CG ASN B 189 79.963 11.314 34.618 1.00 24.95 B
ATOM 2983 OD1 ASN B 189 78.793 10.936 34.720 1.00 28.51 B
ATOM 2984 ND2 ASN B 189 80.984 10.633 35.123 1.00 28.38 B
ATOM 2985 C ASN B 189 79.378 14.131 32.168 1.00 20.22 B
ATOM 2986 0 ASN B 189 79.447 15.010 33.025 1.00 20.71 B
ATOM 2987 N ARG B 190 79.064 14.379 30.901 1.00 18.69 B
ATOM 2988 CA ARG B 190 78.712 15.717 30.445 1.00 17.54 B
ATOM 2989 CB ARG B 190 79.756 16.267 29.473 1.00 18.78 B
ATOM 2990 CG ARG B 190 81.078 16.665 30.112 1.00 20.73 B
ATOM 2991 CD ARG B 190 80.874 17.610 31.293 1.00 20.81 B
ATOM 2992 NE ARG B 190 82.112 18.302 31.637 1.00 21.75 B
ATOM 2993 CZ ARG B 190 82.312 18.981 32.761 1.00 23.62 B
ATOM 2994 NH1 ARG B 190 81.353 19.064 33.673 1.00 23.18 B
ATOM 2995 NH2 ARG B 190 83.475 19.588 32.966 1.00 24.69 B
ATOM 2996 C ARG B 190 77.372 15.587 29.731 1.00 15.48 B
ATOM 2997 0 ARG B 190 77.232 15.972 28.573 1.00 15.70 B
ATOM 2998 N PRO B 191 76.364 15.036 30.423 1.00 15.29 B
ATOM 2999 CD PRO B 191 76.361 14.613 31.837 1.00 15.47 B
ATOM 3000 CA PRO B 191 75.040 14.859 29.825 1.00 14.51 B
ATOM 3001 CB PRO B 191 74.293 14.048 30.877 1.00 13.89 B
ATOM 3002 CG PRO B 191 74.884 14.554 32.161 1.00 16.30 B
ATOM 3003 C PRO B 191 74.309 16.143 29.481 1.00 13.52 B

ATOM 3004 0 PRO B 191 73.562 16.192 28.506 1.00 15.32 B
ATOM 3005 N LEU B 192 74.508 17.181 30.282 1.00 12.45 B
ATOM 3006 CA LEU B 192 73.811 18.431 30.018 1.00 11.68 B
ATOM 3007 CB LEU B 192 73.994 19.402 31.186 1.00 13.81 B
ATOM 3008 CG LEU B 192 73.285 20.750 31.029 1.00 14.64 B
ATOM 3009 CD1 LEU B 192 71.785 20.551 30.853 1.00 15.86 B
ATOM 3010 CD2 LEU B 192 73.568 21.591 32.255 1.00 13.92 B
ATOM 3011 C LEU B 192 74.281 19.071 28.727 1.00 10.06 B
ATOM 3012 0 LEU B 192 73.462 19.467 27.902 1.00 11.66 B
ATOM 3013 N THR B 193 75.595 19.175 28.543 1.00 11.24 B
ATOM 3014 CA THR B 193 76.122 19.775 27.321 1.00 11.61 B
ATOM 3015 CB THR B 193 77.658 19.919 27.378 1.00 12.91 B
ATOM 3016 001 THR B 193 78.012 20.820 28.435 1.00 16.19 B
ATOM 3017 CG2 THR B 193 78.190 20.466 26.060 1.00 13.79 B
ATOM 3018 C THR B 193 75.744 18.926 26.112 1.00 12.24 B
ATOM 3019 0 THR B 193 75.334 19.449 25.071 1.00 13.82 B
ATOM 3020 N CYS B 194 75.866 17.611 26.259 1.00 12.71 B
ATOM 3021 CA CYS B 194 75.540 16.698 25.173 1.00 13.72 B
ATOM 3022 CB CYS B 194 75.856 15.257 25.587 1.00 16.41 B
ATOM 3023 SG CYS B 194 75.559 14.034 24.288 1.00 21.35 B
ATOM 3024 C CYS B 194 74.078 16.791 24.749 1.00 13.33 B
ATOM 3025 0 CYS B 194 73.777 16.995 23.571 1.00 13.97 B
ATOM 3026 N ILE B 195 73.169 16.649 25.709 1.00 13.46 B
ATOM 3027 CA ILE B 195 71.750 16.695 25.392 1.00 13.88 B
ATOM 3028 CB ILE B 195 70.881 16.242 26.604 1.00 15.55 B
ATOM 3029 CG2 ILE B 195 70.542 17.420 27.507 1.00 17.85 B
ATOM 3030 CG1 ILE B 195 69.585 15.610 26.092 1.00 18.57 B
ATOM 3031 CD1 ILE B 195 69.809 14.372 25.243 1.00 19.03 B
ATOM 3032 C ILE B 195 71.310 18.075 24.910 1.00 13.24 B
ATOM 3033 0 ILE B 195 70.450 18.176 24.036 1.00 14.65 B
ATOM 3034 N MET B 196 71.899 19.137 25.456 1.00 11.86 B
ATOM 3035 CA MET B 196 71.522 20.482 25.031 1.00 11.58 B
ATOM 3036 CB MET B 196 72.135 21.544 25.954 1.00 11.62 B
ATOM 3037 CG MET B 196 71.402 21.676 27.292 1.00 13.56 B
ATOM 3038 SD MET B 196 69.755 22.411 27.117 1.00 14.36 B
ATOM 3039 CE MET B 196 70.219 24.137 26.857 1.00 12.74 B
ATOM 3040 C MET B 196 71.959 20.717 23.594 1.00 11.56 B
ATOM 3041 0 MET B 196 71.246 21.349 22.819 1.00 13.66 B
ATOM 3042 N TYR B 197 73.131 20.204 23.232 1.00 12.14 B
ATOM 3043 CA TYR B 197 73.597 20.370 21.865 1.00 13.19 B
ATOM 3044 CB TYR B 197 75.029 19.845 21.704 1.00 16.11 B
ATOM 3045 CG TYR B 197 75.664 20.235 20.384 1.00 19.16 B
ATOM 3046 CD1 TYR B 197 76.023 21.553 20.129 1.00 19.15 B
ATOM 3047 CE1 TYR B 197 76.598 21.922 18.927 1.00 20.63 B
ATOM 3048 CD2 TYR B 197 75.895 19.291 19.396 1.00 21.73 B
ATOM 3049 CE2 TYR B 197 76.473 19.651 18.184 1.00 22.58 B
ATOM 3050 CZ TYR B 197 76.821 20.969 17.961 1.00 22.00 B
ATOM 3051 OH TYR B 197 77.394 21.338 16.766 1.00 25.29 B
ATOM 3052 C TYR B 197 72.660 19.601 20.935 1.00 12.79 B
ATOM 3053 0 TYR B 197 72.290 20.092 19.871 1.00 14.13 B
ATOM 3054 N ALA B 198 72.268 18.397 21.337 1.00 12.79 B
ATOM 3055 CA ALA B 198 71.370 17.589 20.511 1.00 13.49 B
ATOM 3056 CB ALA B 198 71.183 16.202 21.137 1.00 15.13 B
ATOM 3057 C ALA B 198 70.018 18.276 20.323 1.00 13.07 B
ATOM 3058 0 ALA B 198 69.467 18.293 19.219 1.00 14.87 B
ATOM 3059 N ILE B 199 69.490 18.848 21.400 1.00 14.66 B
ATOM 3060 CA ILE B 199 68.208 19.544 21.360 1.00 13.61 B
ATOM 3061 CB ILE B 199 67.773 19.963 22.781 1.00 13.70 B
ATOM 3062 CG2 ILE B 199 66.611 20.943 22.704 1.00 14.17 B
ATOM 3063 CGl ILE B 199 67.415 18.720 23.603 1.00 13.10 B
ATOM 3064 CDl ILE B 199 67.159 19.016 25.079 1.00 15.18 B
ATOM 3065 C ILE B 199 68.277 20.788 20.482 1.00 14.02 B
ATOM 3066 0 ILE B 199 67.387 21.037 19.661 1.00 15.67 B
ATOM 3067 N PHE B 200 69.330 21.577 20.646 1.00 13.42 B
ATOM 3068 CA PHE B 200 69.457 22.782 19.839 1.00 14.38 B
ATOM 3069 CB PHE B 200 70.610 23.638 20.355 1.00 14.15 B
ATOM 3070 CG PHE B 200 70.203 24.591 21.444 1.00 14.10 B

ATOM 3071 CD1 PHE B 200 69.589 24.126 22.595 1.00 14.95 B
ATOM 3072 CD2 PHE B 200 70.420 25.951 21.307 1.00 15.05 B
ATOM 3073 CEl PHE B 200 69.196 25.003 23.592 1.00 16.14 B
ATOM 3074 CE2 PHE B 200 70.030 26.836 22.301 1.00 16.54 B
ATOM 3075 CZ PHE B 200 69.418 26.361 23.443 1.00 15.94 B
ATOM 3076 C PHE B 200 69.615 22.467 18.354 1.00 15.97 B
ATOM 3077 0 PHE B 200 69.102 23.198 17.503 1.00 17.75 B
ATOM 3078 N GLN B 201 70.316 21.382 18.041 1.00 16.45 B
ATOM 3079 CA GLN B 201 70.492 20.975 16.648 1.00 17.39 B
ATOM 3080 CB GLN B 201 71.487 19.814 16.556 1.00 18.95 B
ATOM 3081 CG GLN B 201 72.933 20.219 16.764 1.00 21.95 B
ATOM 3082 CD GLN B 201 73.575 20,741 15.496 1.00 25.50 B
ATOM 3083 OE1 GLN B 201 73.034 21.624 14.827 1.00 27.92 B
ATOM 3084 NE2 GLN B 201 74.738 20.195 15.157 1.00 28.48 B
ATOM 3085 C GLN B 201 69.142 20.530 16.092 1.00 18.10 B
ATOM 3086 0 GLN B 201 68.764 20.889 14.975 1.00 18.77 B
ATOM 3087 N GLU B 202 68.413 19.753 16.890 1.00 16.96 B
ATOM 3088 CA GLU B 202 67.104 19.243 16.494 1.00 18.57 B
ATOM 3089 CB GLU B 202 66.542 18.353 17.614 1.00 19.51 B
ATOM 3090 CG GLU B 202 65.111 17.844 17.418 1.00 21.80 B
ATOM 3091 CD GLU B 202 64.980 16.823 16.302 1.00 26.27 B
ATOM 3092 OE1 GLU B 202 65.882 15.971 16.157 1.00 28.81 B
ATOM 3093 OE2 GLU B 202 63.959 16.859 15.579 1.00 28.69 B
ATOM 3094 C GLU B 202 66.125 20.373 16.189 1.00 18.30 B
ATOM 3095 0 GLU B 202 65.340 20.285 15.243 1.00 19.71 B
ATOM 3096 N ARG B 203 66.172 21.435 16.987 1.00 17.58 B
ATOM 3097 CA ARG B 203 65.267 22.565 16.803 1.00 16.55 B
ATOM 3098 CB ARG B 203 64.840 23.111 18.167 1.00 16.38 B
ATOM 3099 CG ARG B 203 63.924 22.170 18.935 1.00 17.22 B
ATOM 3100 CD ARG B 203 63.510 22.753 20.277 1.00 15.58 B
ATOM 3101 NE ARG B 203 62.545 21.897 20.955 1.00 14.62 B
ATOM 3102 CZ ARG B 203 61.255 21.817 20.635 1.00 15.74 B
ATOM 3103 NH1 ARG B 203 60.763 22.555 19.648 1.00 18.09 B
ATOM 3104 NH2 ARG B 203 60.459 20.976 21.282 1.00 16.38 B
ATOM 3105 C ARG B 203 65.834 23.694 15.946 1.00 15.84 B
ATOM 3106 0 ARG B 203 65.185 24.723 15.759 1.00 16.66 B
ATOM 3107 N ASP B 204 67.040 23.488 15.424 1.00 16.75 B
ATOM 3108 CA ASP B 204 67.716 24.466 14.577 1.00 16.33 B
ATOM 3109 CB ASP B 204 66.920 24.691 13.288 1.00 18.57 B
ATOM 3110 CG ASP B 204 67.782 25.213 12.158 1.00 22.21 B
ATOM 3111 OD1 ASP B 204 67.233 25.841 11.228 1.00 27.95 B
ATOM 3112 OD2 ASP B 204 69.009 24.984 12.193 1.00 25.64 B
ATOM 3113 C ASP B 204 67.903 25.800 15.294 1.00 17.15 B
ATOM 3114 0 ASP B 204 67.976 26.856 14.661 1.00 16.89 B
ATOM 3115 N LEU B 205 67.995 25.749 16.618 1.00 15.64 B
ATOM 3116 CA LEU B 205 68.159 26.963 17.408 1.00 16.23 B
ATOM 3117 CB LEU B 205 67.936 26.656 18.891 1.00 15.88 B
ATOM 3118 CG LEU B 205 66.484 26.412 19.312 1.00 16.04 B
ATOM 3119 CD1 LEU B 205 66.455 25.906 20.749 1.00 17.66 B
ATOM 3120 CD2 LEU B 205 65.680 27.705 19.170 1.00 17.91 B
ATOM 3121 C LEU B 205 69.499 27.671 17.226 1.00 14.59 B
ATOM 3122 0 LEU B 205 69.571 28.895 17.356 1.00 14.90 B
ATOM 3123 N LEU B 206 70.561 26.924 16.932 1.00 15.05 B
ATOM 3124 CA LEU B 206 71.859 27.563 16.743 1.00 13.63 B
ATOM 3125 CB LEU B 206 72.971 26.524 16.569 1.00 14.65 B
ATOM 3126 CG LEU B 206 73.267 25.588 17.741 1.00 17.27 B
ATOM 3127 CD1 LEU B 206 74.465 24.720 17.381 1.00 18.31 B
ATOM 3128 CD2 LEU B 206 73.557 26.388 18.999 1.00 18.18 B
ATOM 3129 C LEU B 206 71.823 28.477 15.521 1.00 14.30 B
ATOM 3130 0 LEU B 206 72.265 29.622 15.576 1.00 15.32 B
ATOM 3131 N LYS B 207 71.280 27.974 14.419 1.00 15.44 B
ATOM 3132 CA LYS B 207 71.196 28.765 13.194 1.00 17.00 B
ATOM 3133 CB LYS B 207 70.775 27.880 12.021 1.00 20.27 B
ATOM 3134 CG LYS B 207 71.628 26.645 11.807 1.00 22.82 B
ATOM 3135 CD LYS B 207 73.060 26.994 11.474 1.00 23.24 B
ATOM 3136 CE LYS B 207 73.611 26.057 10.401 1.00 25.60 B
ATOM 3137 NZ LYS B 207 73.358 24.623 10.717 1.00 27.60 B

ATOM 3138 C LYS B 207 70.192 29.907 13.327 1.00 15.29 B
ATOM 3139 0 LYS B 207 70.448 31.028 12.897 1.00 16.50 B
ATOM 3140 N THR B 208 69.043 29.613 13.922 1.00 15.28 B
ATOM 3141 CA THR B 208 68.001 30.618 14.090 1.00 15.47 B
ATOM 3142 CB THR B 208 66.784 30.030 14.838 1.00 14.55 B
ATOM 3143 001 THR B 208 66.194 28.997 14.041 1.00 17.60 B
ATOM 3144 CG2 THR B 208 65.742 31.108 15.114 1.00 17.27 B
ATOM 3145 C THR B 208 68.477 31.864 14.829 1.00 15.11 B
ATOM 3146 0 THR B 208 68.185 32.989 14.418 1.00 17.42 B
ATOM 3147 N PHE B 209 69.218 31.665 15.916 1.00 15.59 B
ATOM 3148 CA PHE B 209 69.704 32.783 16.715 1.00 15.09 B
ATOM 3149 CB PHE B 209 69.387 32.526 18.189 1.00 15.53 B
ATOM 3150 CG PHE B 209 67.923 32.562 18.491 1.00 13.57 B
ATOM 3151 CD1 PHE B 209 67.234 33.762 18.480 1.00 15.65 B
ATOM 3152 CD2 PHE B 209 67.221 31.394 18.732 1.00 14.27 B
ATOM 3153 CE1 PHE B 209 65.871 33.799 18.702 1.00 16.18 B
ATOM 3154 CE2 PHE B 209 65.856 31.423 18.953 1.00 14.95 B
ATOM 3155 CZ PHE B 209 65.181 32.629 18.938 1.00 16.57 B
ATOM 3156 C PHE B 209 71.188 33.084 16.545 1.00 14.79 B
ATOM 3157 0 PHE B 209 71.777 33.805 17.348 1.00 14.76 B
ATOM 3158 N ARG B 210 71.776 32.531 15.490 1.00 14.65 B
ATOM 3159 CA ARG B 210 73.186 32.724 15.175 1.00 15.09 B
ATOM 3160 CB ARG B 210 73.413 34.124 14.598 1.00 16.89 B
ATOM 3161 CG ARG B 210 72.658 34.367 13.295 1.00 21.59 B
ATOM 3162 CD ARG B 210 73.165 33.451 12.184 1.00 23.89 B
ATOM 3163 NE ARG B 210 72.395 33.593 10.950 1.00 27.05 B
ATOM 3164 CZ ARG B 210 72.737 33.043 9.788 1.00 26.76 B
ATOM 3165 NH1 ARG B 210 73.840 32.314 9.697 1.00 28.04 B
ATOM 3166 NH2 ARG B 210 71.977 33.222 8.715 1.00 26.92 B
ATOM 3167 C ARG B 210 74.117 32.491 16.356 1.00 14.01 B
ATOM 3168 0 ARG B 210 74.960 33.328 16.672 1.00 16.29 B
ATOM 3169 N ILE B 211 73.948 31.345 17.002 1.00 14.62 B
ATOM 3170 CA ILE B 211 74.794 30.961 18.121 1.00 13.55 B
ATOM 3171 CB ILE B 211 74.008 30.211 19.204 1.00 13.69 B
ATOM 3172 CG2 ILE B 211 74.885 30.028 20.436 1.00 12.94 B
ATOM 3173 CG1 ILE B 211 72.746 30.991 19.577 1.00 15.10 B
ATOM 3174 CD1 ILE B 211 71.828 30.235 20.510 1.00 14.24 B
ATOM 3175 C ILE B 211 75.797 29.988 17.515 1.00 13.29 B
ATOM 3176 0 ILE B 211 75.413 28.912 17.057 1.00 14.84 B
ATOM 3177 N SER B 212 77.071 30.366 17.485 1.00 12.75 B
ATOM 3178 CA SER B 212 78.079 29.476 16.930 1.00 13.87 B
ATOM 3179 CB SER B 212 79.441 30.170 16.894 1.00 17.08 B
ATOM 3180 OG SER B 212 79.395 31.331 16.084 1.00 25.78 B
ATOM 3181 C SER B 212 78.162 28.223 17.791 1.00 14.02 B
ATOM 3182 0 SER B 212 77.945 28.279 19.004 1.00 13.97 B
ATOM 3183 N SER B 213 78.467 27.086 17.173 1.00 14.06 B
ATOM 3184 CA SER B 213 78.587 25.860 17.944 1.00 14.77 B
ATOM 3185 CB SER B 213 78.908 24.677 17.034 1.00 14.51 B
ATOM 3186 OG SER B 213 77.763 24.305 16.287 1.00 19.92 B
ATOM 3187 C SER B 213 79.675 26.017 18.998 1.00 13.32 B
ATOM 3188 0 SER B 213 79.532 25.523 20.116 1.00 13.15 B
ATOM 3189 N ASP B 214 80.759 26.709 18.652 1.00 13.09 B
ATOM 3190 CA ASP B 214 81.844 26.903 19.607 1.00 13.63 B
ATOM 3191 CB ASP B 214 83.012 27.666 18.978 1.00 16.13 B
ATOM 3192 CG ASP B 214 84.123 27.934 19.975 1.00 18.90 B
ATOM 3193 001 ASP B 214 84.774 26.964 20.420 1.00 22.48 B
ATOM 3194 OD2 ASP B 214 84.336 29.113 20.323 1.00 22.40 B
ATOM 3195 C ASP B 214 81.371 27.662 20.835 1.00 12.29 B
ATOM 3196 0 ASP B 214 81.677 27.271 21.961 1.00 14.25 B
ATOM 3197 N THR B 215 80.621 28.742 20.622 1.00 11.07 B
ATOM 3198 CA THR B 215 80.125 29.542 21.741 1.00 11.42 B
ATOM 3199 CB THR B 215 79.372 30.786 21.249 1.00 11.51 B
ATOM 3200 001 THR B 215 80.233 31.547 20.400 1.00 14.55 B
ATOM 3201 CG2 THR B 215 78.936 31.656 22.424 1.00 13.98 B
ATOM 3202 C THR B 215 79.168 28.728 22.584 1.00 9.02 B
ATOM 3203 0 THR B 215 79.232 28.741 23.810 1.00 10.90 B
ATOM 3204 N PHE B 216 78.267 28.014 21.923 1.00 10.45 B

ATOM 3205 CA PHE B 216 77.296 27.213 22.647 1.00 8.91 B
ATOM 3206 CB PHE B 216 76.332 26.533 21.681 1.00 12.68 B
ATOM 3207 CG PHE B 216 75.185 25.860 22.366 1.00 16.02 B
ATOM 3208 CD1 PHE B 216 74.181 26.614 22.953 1.00 17.00 B
ATOM 3209 CD2 PHE B 216 75.121 24.483 22.452 1.00 17.44 B
ATOM 3210 CE1 PHE B 216 73.131 26.004 23.617 1.00 18.37 B
ATOM 3211 CE2 PHE B 216 74.072 23.868 23.115 1.00 17.00 B
ATOM 3212 CZ PHE B 216 73.081 24.630 23.695 1.00 15.22 B
ATOM 3213 C PHE B 216 77.965 26.149 23.505 1.00 9.66 B
ATOM 3214 0 PHE B 216 77.617 25.970 24.667 1.00 11.34 B
ATOM 3215 N ILE B 217 78.927 25.438 22.926 1.00 9.00 B
ATOM 3216 CA ILE B 217 79.620 24.397 23.663 1.00 9.44 B
ATOM 3217 CB ILE B 217 80.524 23.569 22.730 1.00 12.02 B
ATOM 3218 CG2 ILE B 217 81.324 22.551 23.542 1.00 13.95 B
ATOM 3219 CG1 ILE B 217 79.653 22.843 21.699 1.00 14.93 B
ATOM 3220 CD1 ILE B 217 80.434 22.217 20.552 1.00 17.33 B
ATOM 3221 C ILE B 217 80.444 24.976 24.811 1.00 9.65 B
ATOM 3222 0 ILE B 217 80.468 24.410 25.909 1.00 11.30 B
ATOM 3223 N THR B 218 81.096 26.109 24.575 1.00 9.00 B
ATOM 3224 CA THR B 218 81.902 26.728 25.617 1.00 9.70 B
ATOM 3225 CB THR B 218 82.638 27.971 25.082 1.00 10.90 B
ATOM 3226 OG1 THR B 218 83.504 27.581 24.010 1.00 13.11 B
ATOM 3227 CG2 THR B 218 83.460 28.624 26.183 1.00 12.91 B
ATOM 3228 C THR B 218 81.018 27.121 26.794 1.00 7.92 B
ATOM 3229 0 THR B 218 81.350 26.844 27.950 1.00 9.92 B
ATOM 3230 N TYR B 219 79.885 27.754 26.508 1.00 8.85 B
ATOM 3231 CA TYR B 219 78.969 28.153 27.575 1.00 7.47 B
ATOM 3232 CB TYR B 219 77.785 28.973 27.034 1.00 8.03 B
ATOM 3233 CG TYR B 219 76.761 29.259 28.120 1.00 9.13 B
ATOM 3234 CD1 TYR B 219 76.955 30.288 29.035 1.00 11.36 B
ATOM 3235 CE1 TYR B 219 76.068 30.488 30.091 1.00 11.08 B
ATOM 3236 CD2 TYR B 219 75.647 28.438 28.287 1.00 11.17 B
ATOM 3237 CE2 TYR B 219 74.759 28.627 29.337 1.00 10.37 B
ATOM 3238 CZ TYR B 219 74.975 29.652 30.235 1.00 9.68 B
ATOM 3239 OH TYR B 219 74.103 29.831 31.286 1.00 10.38 B
ATOM 3240 C TYR B 219 78.408 26.935 28.299 1.00 8.12 B
ATOM 3241 0 TYR B 219 78.413 26.883 29.526 1.00 11.39 B
ATOM 3242 N MET B 220 77.923 25.952 27.547 1.00 8.60 B
ATOM 3243 CA MET B 220 77.342 24.771 28.178 1.00 9.02 B
ATOM 3244 CB MET B 220 76.727 23.847 27.123 1.00 10.53 B
ATOM 3245 CG MET B 220 75.423 24.381 26.515 1.00 14.14 B
ATOM 3246 SD MET B 220 74.151 24.829 27.748 1.00 13.79 B
ATOM 3247 CE MET B 220 74.167 23.361 28.781 1.00 18.84 B
ATOM 3248 C MET B 220 78.327 24.004 29.050 1.00 8.93 B
ATOM 3249 0 MET B 220 77.965 23.557 30.135 1.00 10.27 B
ATOM 3250 N MET B 221 79.569 23.869 28.587 1.00 10.22 B
ATOM 3251 CA MET B 221 80.592 23.170 29.364 1.00 10.08 B
ATOM 3252 CB MET B 221 81.863 22.989 28.528 1.00 12.14 B
ATOM 3253 CG MET B 221 81.739 21.903 27.468 1.00 14.95 B
ATOM 3254 SD MET B 221 81.567 20.264 28.205 1.00 19.05 B
ATOM 3255 CE MET B 221 81.815 19.218 26.735 1.00 19.61 B
ATOM 3256 C MET B 221 80.905 23.955 30.636 1.00 9.04 B
ATOM 3257 0 MET B 221 81.157 23.369 31.693 1.00 12.49 B
ATOM 3258 N THR B 222 80.889 25.279 30.529 1.00 9.15 B
ATOM 3259 CA THR B 222 81.141 26.143 31.673 1.00 9.03 B
ATOM 3260 CB THR B 222 81.285 27.617 31.211 1.00 8.98 B
ATOM 3261 OG1 THR B 222 82.397 27.723 30.312 1.00 11.02 B
ATOM 3262 CG2 THR B 222 81.515 28.538 32.396 1.00 10.19 B
ATOM 3263 C THR B 222 79.972 25.999 32.659 1.00 9.26 B
ATOM 3264 0 THR B 222 80.174 25.817 33.864 1.00 9.87 B
ATOM 3265 N LEU B 223 78.748 26.055 32.141 1.00 10.23 B
ATOM 3266 CA LEU B 223 77.565 25.913 32.987 1.00 8.74 B
ATOM 3267 CB LEU B 223 76.286 26.047 32.149 1.00 9.44 B
ATOM 3268 CG LEU B 223 74.963 25.876 32.908 1.00 10.37 B
ATOM 3269 CD1 LEU B 223 74.679 27.117 33.738 1.00 13.51 B
ATOM 3270 CD2 LEU B 223 73.823 25.633 31.922 1.00 10.73 B
ATOM 3271 C LEU B 223 77.570 24.553 33.676 1.00 9.68 B

ATOM 3272 0 LEU B 223 77.361 24.464 34.890 1.00 10.59 B
ATOM 3273 N GLU B 224 77.820 23.498 32.904 1.00 11.60 B
ATOM 3274 CA GLU B 224 77.821 22.153 33.461 1.00 12.62 B
ATOM 3275 CB GLU B 224 78.054 21.109 32.362 1.00 14.95 B
ATOM 3276 CG GLU B 224 77.913 19.669 32.858 1.00 15.75 B
ATOM 3277 CD GLU B 224 77.398 18.713 31.794 1.00 16.89 B
ATOM 3278 OE1 GLU B 224 77.634 18.964 30.592 1.00 15.87 B
ATOM 3279 OE2 GLU B 224 76.763 17.698 32.160 1.00 14.54 B
ATOM 3280 C GLU B 224 78.856 22.008 34.567 1.00 12.74 B
ATOM 3281 0 GLU B 224 78.618 21.316 35.558 1.00 15.06 B
ATOM 3282 N ASP B 225 79.998 22.673 34.409 1.00 12.14 B
ATOM 3283 CA ASP B 225 81.048 22.609 35.421 1.00 14.65 B
ATOM 3284 CB ASP B 225 82.337 23.261 34.921 1.00 18.41 B
ATOM 3285 CG ASP B 225 83.400 22.247 34.575 1.00 23.04 B
ATOM 3286 OD1 ASP B 225 83.409 21.168 35.205 1.00 24.72 B
ATOM 3287 OD2 ASP B 225 84.234 22.536 33.691 1.00 24.18 B
ATOM 3288 C ASP B 225 80.639 23.298 36.709 1.00 13.84 B
ATOM 3289 0 ASP B 225 81.281 23.109 37.744 1.00 15.90 B
ATOM 3290 N HIS B 226 79.587 24.109 36.650 1.00 12.52 B
ATOM 3291 CA HIS B 226 79.121 24.803 37.844 1.00 12.52 B
ATOM 3292 CB HIS B 226 78.644 26.214 37.501 1.00 13.34 B
ATOM 3293 CG HIS B 226 79.770 27.188 37.348 1.00 16.73 B
ATOM 3294 CD2 HIS B 226 80.178 28.211 38.135 1.00 18.20 B
ATOM 3295 ND1 HIS B 226 80.683 27.108 36.318 1.00 18.23 B
ATOM 3296 CE1 HIS B 226 81.609 28.037 36.481 1.00 19.31 B
ATOM 3297 NE2 HIS B 226 81.327 28.719 37.576 1.00 20.80 B
ATOM 3298 C HIS B 226 78.066 24.034 38.620 1.00 11.11 B
ATOM 3299 0 HIS B 226 77.548 24.512 39.631 1.00 13.59 B
ATOM 3300 N TYR B 227 77.731 22.848 38.124 1.00 11.83 B
ATOM 3301 CA TYR B 227 76.823 21.965 38.844 1.00 11.89 B
ATOM 3302 CB TYR B 227 76.035 21.059 37.892 1.00 11.76 B
ATOM 3303 CG TYR B 227 74.735 21.667 37.415 1.00 7.86 B
ATOM 3304 CD1 TYR B 227 74.689 22.487 36.289 1.00 11.31 B
ATOM 3305 CE1 TYR B 227 73.498 23.060 35.869 1.00 11.23 B
ATOM 3306 CD2 TYR B 227 73.552 21.439 38.106 1.00 7.51 B
ATOM 3307 CE2 TYR B 227 72.357 22.011 37.693 1.00 9.51 B
ATOM 3308 CZ TYR B 227 72.339 22.821 36.572 1.00 9.15 B
ATOM 3309 OH TYR B 227 71.151 23.395 36.154 1.00 11.04 B
ATOM 3310 C TYR B 227 77.800 21.129 39.679 1.00 14.10 B
ATOM 3311 0 TYR B 227 78.899 20.827 39.220 1.00 16.43 B
ATOM 3312 N HIS B 228 77.421 20.777 40.903 1.00 14.97 B
ATOM 3313 CA HIS B 228 78.304 19.994 41.769 1.00 16.33 B
ATOM 3314 CB HIS B 228 77.999 20.269 43.244 1.00 18.10 B
ATOM 3315 CG HIS B 228 78.094 21.711 43.635 1.00 21.87 B
ATOM 3316 CD2 HIS B 228 78.179 22.837 42.889 1.00 23.53 B
ATOM 3317 ND1 HIS B 228 78.089 22.122 44.951 1.00 24.44 B
ATOM 3318 CE1 HIS B 228 78.169 23.439 44.999 1.00 24.66 B
ATOM 3319 NE2 HIS B 228 78.225 23.898 43.761 1.00 24.22 B
ATOM 3320 C HIS B 228 78.131 18.503 41.513 1.00 15.58 B
ATOM 3321 0 HIS B 228 77.074 17.945 41.802 1.00 17.45 B
ATOM 3322 N SER B 229 79.165 17.845 41.000 1.00 17.16 B
ATOM 3323 CA SER B 229 79.055 16.421 40.713 1.00 18.30 B
ATOM 3324 CB SER B 229 80.221 15.964 39.826 1.00 19.82 B
ATOM 3325 OG SER B 229 81.474 16.185 40.448 1.00 23.03 B
ATOM 3326 C SER B 229 78.983 15.565 41.978 1.00 19.26 B
ATOM 3327 0 SER B 229 78.559 14.412 41.928 1.00 22.10 B
ATOM 3328 N ASP B 230 79.379 16.141 43.110 1.00 18.39 B
ATOM 3329 CA ASP B 230 79.360 15.436 44.390 1.00 20.29 B
ATOM 3330 CB ASP B 230 80.375 16.068 45.351 1.00 23.12 B
ATOM 3331 CG ASP B 230 80.154 17.559 45.536 1.00 26.32 B
ATOM 3332 OD1 ASP B 230 80.282 18.307 44.545 1.00 29.20 B
ATOM 3333 OD2 ASP B 230 79.851 17.986 46.673 1.00 31.52 B
ATOM 3334 C ASP B 230 77.973 15.432 45.032 1.00 19.11 B
ATOM 3335 0 ASP B 230 77.712 14.657 45.956 1.00 21.35 B
ATOM 3336 N VAL B 231 77.086 16.303 44.552 1.00 16.59 B
ATOM 3337 CA VAL B 231 75.723 16.367 45.075 1.00 15.74 B
ATOM 3338 CB VAL B 231 75.062 17.715 44.713 1.00 16.21 B

ATOM 3339 CG1 VAL B 231 73.601 17.717 45.111 1.00 16.50 B
ATOM 3340 CG2 VAL B 231 75.793 18.846 45.432 1.00 16.71 B
ATOM 3341 C VAL B 231 74.981 15.198 44.424 1.00 15.11 B
ATOM 3342 0 VAL B 231 75.015 15.037 43.212 1.00 16.08 B
ATOM 3343 N ALA B 232 74.316 14.383 45.237 1.00 14.78 B
ATOM 3344 CA ALA B 232 73.630 13.190 44.745 1.00 14.78 B
ATOM 3345 CB ALA B 232 73.053 12.410 45.925 1.00 17.07 B
ATOM 3346 C ALA B 232 72.551 13.349 43.673 1.00 12.72 B
ATOM 3347 0 ALA B 232 72.533 12.598 42.694 1.00 14.91 B
ATOM 3348 N TYR B 233 71.653 14.308 43.852 1.00 13.10 B
ATOM 3349 CA TYR B 233 70.571 14.493 42.892 1.00 10.73 B
ATOM 3350 CB TYR B 233 69.228 14.513 43.632 1.00 12.23 B
ATOM 3351 CG TYR B 233 68.015 14.694 42.744 1.00 12.27 B
ATOM 3352 CD1 TYR B 233 67.359 13.598 42.200 1.00 13.49 B
ATOM 3353 CE1 TYR B 233 66.239 13.756 41.407 1.00 13.18 B
ATOM 3354 CD2 TYR B 233 67.519 15.959 42.464 1.00 11.96 B
ATOM 3355 CE2 TYR B 233 66.398 16.129 41.666 1.00 12.76 B
ATOM 3356 CZ TYR B 233 65.763 15.023 41.145 1.00 11.14 B
ATOM 3357 OH TYR B 233 64.639 15.179 40.371 1.00 12.48 B
ATOM 3358 C TYR B 233 70.698 15.752 42.038 1.00 10.96 B
ATOM 3359 0 TYR B 233 70.626 15.681 40.813 1.00 12.54 B
ATOM 3360 N HIS B 234 70.885 16.895 42.687 1.00 11.58 B
ATOM 3361 CA HIS B 234 70.981 18.160 41.972 1.00 11.81 B
ATOM 3362 CB HIS B 234 70.614 19.311 42.909 1.00 12.44 B
ATOM 3363 CG HIS B 234 69.167 19.330 43.286 1.00 11.54 B
ATOM 3364 CD2 HIS B 234 68.069 19.672 42.570 1.00 10.42 B
ATOM 3365 ND1 HIS B 234 68.710 18.933 44.523 1.00 13.88 B
ATOM 3366 CE1 HIS B 234 67.392 19.029 44.554 1.00 12.19 B
ATOM 3367 NE2 HIS B 234 66.979 19.474 43.381 1.00 12.46 B
ATOM 3368 C HIS B 234 72.306 18.446 41.279 1.00 12.18 B
ATOM 3369 0 HIS B 234 72.941 19.473 41.521 1.00 14.48 B
ATOM 3370 N ASN B 235 72.710 17.525 40.412 1.00 11.91 B
ATOM 3371 CA ASN B 235 73.933 17.665 39.634 1.00 11.80 B
ATOM 3372 CB ASN B 235 74.814 16.423 39.795 1.00 13.03 B
ATOM 3373 CG ASN B 235 74.045 15.137 39.605 1.00 15.08 B
ATOM 3374 ODl ASN B 235 73.464 14.910 38.550 1.00 16.15 B
ATOM 3375 ND2 ASN B 235 74.033 14.288 40.632 1.00 14.23 B
ATOM 3376 C ASN B 235 73.499 17.859 38.179 1.00 11.93 B
ATOM 3377 0 ASN B 235 72.302 17.952 37.907 1.00 12.50 B
ATOM 3378 N SER B 236 74.447 17.914 37.247 1.00 11.66 B
ATOM 3379 CA SER B 236 74.076 18.159 35.851 1.00 10.65 B
ATOM 3380 CB SER B 236 75.323 18.424 35.000 1.00 11.10 B
ATOM 3381 OG SER B 236 76.040 17.240 34.710 1.00 15.22 B
ATOM 3382 C SER B 236 73.197 17.101 35.177 1.00 10.03 B
ATOM 3383 0 SER B 236 72.570 17.390 34.155 1.00 11.40 B
ATOM 3384 N LEU B 237 73.136 15.892 35.731 1.00 10.91 B
ATOM 3385 CA LEU B 237 72.288 14.860 35.136 1.00 12.41 B
ATOM 3386 CB LEU B 237 72.541 13.491 35.783 1.00 16.51 B
ATOM 3387 CG LEU B 237 71.830 12.326 35.078 1.00 19.49 B
ATOM 3388 CD1 LEU B 237 72.397 12.165 33.674 1.00 20.59 B
ATOM 3389 CD2 LEU B 237 72.012 11.035 35.868 1.00 23,14 B
ATOM 3390 C LEU B 237 70.823 15.260 35.313 1.00 11.44 B
ATOM 3391 0 LEU B 237 69.999 15.046 34.418 1.00 13.26 B
ATOM 3392 N HIS B 238 70.490 15.834 36.468 1.00 11.16 B
ATOM 3393 CA HIS B 238 69.119 16.278 36.716 1.00 11.09 B
ATOM 3394 CB HIS B 238 68.958 16.735 38.173 1.00 10.62 B
ATOM 3395 CG HIS B 238 67.696 17.500 38.436 1.00 10.96 B
ATOM 3396 CD2 HIS B 238 67.503 18.777 38.840 1.00 12.08 B
ATOM 3397 ND1 HIS B 238 66.438 16.958 38.270 1.00 9.61 B
ATOM 3398 CE1 HIS B 238 65.527 17.869 38.558 1.00 11.19 B
ATOM 3399 NE2 HIS B 238 66.147 18.982 38.905 1.00 11.36 B
ATOM 3400 C HIS B 238 68.770 17.417 35.759 1.00 9.75 B
ATOM 3401 0 HIS B 238 67.681 17.447 35.188 1.00 12.78 B
ATOM 3402 N ALA B 239 69.698 18.353 35.580 1.00 9.80 B
ATOM 3403 CA ALA B 239 69.462 19.473 34.670 1.00 8.47 B
ATOM 3404 CB ALA B 239 70.647 20.453 34.713 1.00 9.70 B
ATOM 3405 C ALA B 239 69.253 18.958 33.241 1.00 8.87 B

ATOM 3406 0 ALA B 239 68.380 19.441 32.526 1.00 10.68 B
ATOM 3407 N ALA B 240 70.062 17.988 32.831 1.00 7.68 B
ATOM 3408 CA ALA B 240 69.939 17.416 31.487 1.00 8.79 B
ATOM 3409 CB ALA B 240 71.031 16.393 31.262 1.00 9.79 B
ATOM 3410 C ALA B 240 68.570 16.762 31.336 1.00 9.08 B
ATOM 3411 0 ALA B 240 67.928 16.861 30.288 1.00 10.69 B
ATOM 3412 N ASP B 241 68.128 16.096 32.397 1.00 9.20 B
ATOM 3413 CA ASP B 241 66,837 15.428 32.402 1.00 10.07 B
ATOM 3414 CB ASP B 241 66.688 14.630 33.696 1.00 11.27 B
ATOM 3415 CG ASP B 241 65.346 13.950 33.807 1.00 14.16 B
ATOM 3416 OD1 ASP B 241 64.935 13.286 32.829 1.00 15.44 B
ATOM 3417 OD2 ASP B 241 64.710 14.077 34.872 1.00 16.88 B
ATOM 3418 C ASP B 241 65.693 16.432 32.259 1.00 9.75 B
ATOM 3419 0 ASP B 241 64.779 16.235 31.451 1.00 12.81 B
ATOM 3420 N VAL B 242 65.741 17.509 33.035 1.00 10.84 B
ATOM 3421 CA VAL B 242 64.698 18.526 32.969 1.00 9.51 B
ATOM 3422 CB VAL B 242 64.868 19.557 34.114 1.00 10.60 B
ATOM 3423 CG1 VAL B 242 63.853 20.689 33.979 1.00 11.72 B
ATOM 3424 CG2 VAL B 242 64.683 18.852 35.454 1.00 12.98 B
ATOM 3425 C VAL B 242 64.689 19.210 31.594 1.00 9.02 B
ATOM 3426 0 VAL B 242 63.619 19.481 31.037 1.00 12.09 B
ATOM 3427 N ALA B 243 65.869 19.479 31.043 1.00 8.32 B
ATOM 3428 CA ALA B 243 65.959 20.101 29.723 1.00 8.47 B
ATOM 3429 CB ALA B 243 67.405 20.430 29.388 1.00 11.98 B
ATOM 3430 C ALA B 243 65.368 19.203 28.638 1.00 10.40 B
ATOM 3431 0 ALA B 243 64.612 19.671 27.786 1.00 11.59 B
ATOM 3432 N GLN B 244 65.714 17.917 28.665 1.00 11.84 B
ATOM 3433 CA GLN B 244 65.205 16.981 27.665 1.00 11.95 B
ATOM 3434 CB GLN B 244 65.939 15.641 27.794 1.00 13.74 B
ATOM 3435 CG GLN B 244 65.692 14.656 26.655 1.00 14.26 B
ATOM 3436 CD GLN B 244 64.497 13.765 26.899 1.00 14.50 B
ATOM 3437 OE1 GLN B 244 64.195 13.415 28.038 1.00 15.47 B
ATOM 3438 NE2 GLN B 244 63.822 13.367 25.823 1.00 14.81 B
ATOM 3439 C GLN B 244 63.692 16.790 27.812 1.00 10.88 B
ATOM 3440 0 GLN B 244 62,971 16.686 26.816 1.00 13.33 B
ATOM 3441 N SER B 245 63.212 16.768 29.052 1.00 11.10 B
ATOM 3442 CA SER B 245 61.787 16.605 29.314 1.00 10.90 B
ATOM 3443 CB SER B 245 61.532 16.407 30.809 1.00 11.12 B
ATOM 3444 OG SER B 245 62.145 15.218 31.271 1.00 13.86 B
ATOM 3445 C SER B 245 61,027 17.828 28.830 1.00 11.68 B
ATOM 3446 0 SER B 245 59.923 17.715 28.297 1.00 13.98 B
ATOM 3447 N THR B 246 61.619 19.001 29.031 1.00 12.22 B
ATOM 3448 CA THR B 246 61.006 20.245 28.593 1.00 12.39 B
ATOM 3449 CB THR B 246 61.841 21.454 29.065 1.00 12.58 B
ATOM 3450 OG1 THR B 246 61.779 21.536 30.493 1.00 13.59 B
ATOM 3451 CG2 THR B 246 61.307 22.744 28.471 1.00 14.72 B
ATOM 3452 C THR B 246 60.923 20.223 27.066 1.00 12.97 B
ATOM 3453 0 THR B 246 59.929 20.654 26.478 1.00 14.54 B
ATOM 3454 N HIS B 247 61.971 19.703 26.437 1.00 11.92 B
ATOM 3455 CA HIS B 247 62.027 19.593 24.984 1.00 12.49 B
ATOM 3456 CB HIS B 247 63.370 18.977 24.576 1.00 13.03 B
ATOM 3457 CG HIS B 247 63.425 18.521 23.153 1.00 13.55 B
ATOM 3458 CD2 HIS B 247 63.563 17.283 22.624 1.00 13.60 B
ATOM 3459 ND1 HIS B 247 63.324 19.386 22.085 1.00 14.08 B
ATOM 3460 CE1 HIS B 247 63.398 18.700 20.958 1.00 14.38 B
ATOM 3461 NE2 HIS B 247 63.541 17.421 21.257 1.00 15.75 B
ATOM 3462 C HIS B 247 60.862 18.741 24.479 1.00 13.03 B
ATOM 3463 0 HIS B 247 60.205 19.087 23.497 1.00 13.30 B
ATOM 3464 N VAL B 248 60.592 17.633 25.159 1.00 12.52 B
ATOM 3465 CA VAL B 248 59.490 16.771 24.748 1.00 12.82 B
ATOM 3466 CB VAL B 248 59.519 15.432 25.512 1.00 14.39 B
ATOM 3467 CGl VAL B 248 58.285 14.613 25.174 1.00 17.07 B
ATOM 3468 CG2 VAL B 248 60.774 14.658 25.139 1.00 14.54 B
ATOM 3469 C VAL B 248 58.139 17.455 24.974 1.00 12.57 B
ATOM 3470 0 VAL B 248 57.266 17.429 24.105 1.00 14.95 B
ATOM 3471 N LEU B 249 57.963 18.074 26.136 1.00 14.53 B
ATOM 3472 CA LEU B 249 56.703 18.752 26.437 1.00 15.02 B

ATOM 3473 CB LEU B 249 56.728 19.314 27.861 1.00 17.50 B
ATOM 3474 CG LEU B 249 56.742 18.255 28.969 1.00 16.08 B
ATOM 3475 CD1 LEU B 249 56.855 18.931 30.321 1.00 17.15 B
ATOM 3476 CD2 LEU B 249 55.480 17.410 28.900 1.00 19.85 B
ATOM 3477 C LEU B 249 56.387 19.867 25.439 1.00 15.17 B
ATOM 3478 0 LEU B 249 55.228 20.085 25.090 1.00 17.12 B
ATOM 3479 N LEU B 250 57.416 20.571 24.980 1.00 16.33 B
ATOM 3480 CA LEU B 250 57.218 21.649 24.017 1.00 16.67 B
ATOM 3481 CB LEU B 250 58.530 22.400 23.780 1.00 17.65 B
ATOM 3482 CG LEU B 250 58.951 23.425 24.832 1.00 18.41 B
ATOM 3483 CD1 LEU B 250 60.403 23.821 24.611 1.00 19.35 B
ATOM 3484 CD2 LEU B 250 58.040 24.644 24.750 1,00 18.88 B
ATOM 3485 C LEU B 250 56.696 21.126 22.681 1.00 17.21 B
ATOM 3486 0 LEU B 250 56.080 21.873 21.920 1.00 19.20 B
ATOM 3487 N SER B 251 56.932 19.847 22.403 1.00 18.15 B
ATOM 3488 CA SER B 251 56.499 19.253 21.142 1.00 19.87 B
ATOM 3489 CB SER B 251 57.522 18.215 20.669 1.00 21.48 B
ATOM 3490 OG SER B 251 58.756 18.826 20.328 1.00 27.10 B
ATOM 3491 C SER B 251 55.112 18.618 21.162 1.00 20.31 B
ATOM 3492 0 SER B 251 54.655 18.117 20.135 1.00 20.88 B
ATOM 3493 N THR B 252 54.445 18.639 22.312 1.00 19.49 B
ATOM 3494 CA THR B 252 53.108 18.058 22.421 1.00 20.53 B
ATOM 3495 CB THR B 252 52.479 18.349 23.797 1.00 21.36 B
ATOM 3496 OG1 THR B 252 52.292 19.760 23.949 1.00 26.25 B
ATOM 3497 CG2 THR B 252 53.382 17.846 24.907 1.00 22.53 B
ATOM 3498 C THR B 252 52.217 18.659 21.336 1.00 20.15 B
ATOM 3499 0 THR B 252 52.245 19.864 21.099 1.00 20.80 B
ATOM 3500 N PRO B 253 51.418 17.823 20.658 1.00 21.10 B
ATOM 3501 CD PRO B 253 51.260 16.373 20.863 1.00 21.07 B
ATOM 3502 CA PRO B 253 50.526 18.295 19.593 1.00 20.74 B
ATOM 3503 CB PRO B 253 49.643 17.079 19.329 1.00 21.86 B
ATOM 3504 CG PRO B 253 50.575 15.945 19.581 1.00 21.58 B
ATOM 3505 C PRO B 253 49.712 19.535 19.957 1.,00 21.53 B
ATOM 3506 0 PRO B 253 49.552 20.443 19.142 1.00 22.03 B
ATOM 3507 N ALA B 254 49.210 19.573 21.185 1.00 20.69 B
ATOM 3508 CA ALA B 254 48.402 20.693 21.650 1.00 21.13 B
ATOM 3509 CB ALA B 254 47.831 20.379 23.030 1.00 21.75 B
ATOM 3510 C ALA B 254 49.152 22.024 21.688 1.00 21.24 B
ATOM 3511 0 ALA B 254 48.536 23.087 21.714 1.00 22.02 B
ATOM 3512 N LEU B 255 50.479 21.972 21.694 1.00 21.50 B
ATOM 3513 CA LEU B 255 51.264 23.200 21.736 1.00 20.88 B
ATOM 3514 CB LEU B 255 52.281 23.125 22.874 1.00 19.80 B
ATOM 3515 CG LEU B 255 51.654 22.886 24.248 1.00 19.98 B
ATOM 3516 CD1 LEU B 255 52.740 22.640 25.277 1.00 22.12 B
ATOM 3517 CD2 LEU B 255 50.806 24.091 24.633 1.00 17.78 B
ATOM 3518 C LEU B 255 51.981 23.449 20.417 1.00 22.61 B
ATOM 3519 0 LEU B 255 52.959 24.195 20.362 1.00 23.66 B
ATOM 3520 N ASP B 256 51.476 22.828 19.357 1.00 23.28 B
ATOM 3521 CA ASP B 256 52.057 22.956 18.025 1.00 24.28 B
ATOM 3522 CB ASP B 256 51.319 22.022 17.059 1.00 25.93 B
ATOM 3523 CG ASP B 256 51.996 21.918 15.710 1.00 27.96 B
ATOM 3524 OD1 ASP B 256 53.190 21.546 15.668 1.00 31.17 B
ATOM 3525 OD2 ASP B 256 51.333 22.201 14.689 1.00 31.35 B
ATOM 3526 C ASP B 256 51.991 24.397 17.520 1.00 24.79 B
ATOM 3527 0 ASP B 256 50.913 24.985 17.426 1.00 24.58 B
ATOM 3528 N ALA B 257 53.155 24.958 17.205 1.00 24.40 B
ATOM 3529 CA ALA B 257 53.261 26.325 16.699 1.00 25.31 B
ATOM 3530 CB ALA B 257 52.530 26.442 15.356 1.00 25.56 B
ATOM 3531 C ALA B 257 52.737 27.380 17.670 1.00 25.05 B
ATOM 3532 0 ALA B 257 52.572 28.543 17.297 1.00 27.16 B
ATOM 3533 N VAL B 258 52.488 26.981 18.914 1.00 23.37 B
ATOM 3534 CA VAL B 258 51.980 27.904 19.924 1.00 21.82 B
ATOM 3535 CB VAL B 258 51.441 27.148 21.156 1.00 22.18 B
ATOM 3536 CG1 VAL B 258 51.170 28.127 22.288 1.00 22.06 B
ATOM 3537 CG2 VAL B 258 50.172 26.394 20.791 1.00 21.84 B
ATOM 3538 C VAL B 258 53.032 28.891 20.415 1.00 20.88 B
ATOM 3539 0 VAL B 258 52,732 30.056 20.656 1.00 22.53 B

ATOM 3540 N PHE B 259 54.265 28.421 20.570 1.00 19.20 B
ATOM 3541 CA PHE B 259 55.337 29.278 21.060 1.00 18.21 B
ATOM 3542 CB PHE B 259 56.087 28.561 22.184 1.00 18.51 B
ATOM 3543 CG PHE B 259 55.224 28.221 23.361 1.00 16.16 B
ATOM 3544 CDl PHE B 259 54.723 29.220 24.179 1.00 17.57 B
ATOM 3545 CD2 PHE B 259 54.893 26.905 23.639 1.00 16.17 B
ATOM 3546 CE1 PHE B 259 53.906 28.913 25.252 1.00 18.74 B
ATOM 3547 CE2 PHE B 259 54.077 26.592 24.710 1.00 16.38 B
ATOM 3548 CZ PHE B 259 53.583 27.595 25.517 1.00 20.21 B
ATOM 3549 C PHE B 259 56.325 29.704 19.983 1.00 17.88 B
ATOM 3550 0 PHE B 259 56.598 28.961 19.049 1.00 19.56 B
ATOM 3551 N THR B 260 56.858 30.912 20.128 1.00 17.99 B
ATOM 3552 CA THR B 260 57.837 31,442 19.185 1.00 16.82 B
ATOM 3553 CB THR B 260 58.041 32.949 19.376 1.00 16.92 B
ATOM 3554 OG1 THR B 260 58.521 33.188 20.703 1.00 18.74 B
ATOM 3555 CG2 THR B 260 56.739 33.705 19.160 1.00 19.40 B
ATOM 3556 C THR B 260 59.184 30.771 19.440 1.00 15.28 B
ATOM 3557 0 THR B 260 59.373 30.124 20.467 1.00 15.52 B
ATOM 3558 N ASP B 261 60.123 30.936 18.515 1.00 15.66 B
ATOM 3559 CA ASP B 261 61.444 30.341 18.693 1.00 14.73 B
ATOM 3560 CB ASP B 261 62.339 30.607 17.477 1.00 18.37 B
ATOM 3561 CG ASP B 261 61.789 30.000 16.203 1.00 23.43 B
ATOM 3562 OD1 ASP B 261 61.251 28.874 16.267 1.00 24.71 B
ATOM 3563 OD2 ASP B 261 61.908 30.643 15.136 1.00 24.78 B
ATOM 3564 C ASP B 261 62.114 30.912 19.939 1.00 13.61 B
ATOM 3565 0 ASP B 261 62.843 30.209 20.635 1.00 15.14 B
ATOM 3566 N LEU B 262 61.864 32.187 20.218 1.00 13.37 B
ATOM 3567 CA LEU B 262 62.458 32.839 21.381 1.00 13.88 B
ATOM 3568 CB LEU B 262 62.159 34.340 21.363 1.00 13.41 B
ATOM 3569 CG LEU B 262 62.871 35.171 22.439 1.00 17.34 B
ATOM 3570 CD1 LEU B 262 64.375 35.104 22.217 1.00 17.39 B
ATOM 3571 CD2 LEU B 262 62.389 36.613 22.388 1.00 19.73 B
ATOM 3572 C LEU B 262 61.935 32.232 22.677 1.00 12.72 B
ATOM 3573 0 LEU B 262 62.694 32.045 23.630 1.00 14.39 B
ATOM 3574 N GLU B 263 60.640 31.930 22.716 1.00 13.10 B
ATOM 3575 CA GLU B 263 60.032 31.345 23.902 1.00 13.47 B
ATOM 3576 CB GLU B 263 58.502 31.374 23.772 1.00 15.74 B
ATOM 3577 CG GLU B 263 57.949 32.802 23.714 1.00 16.99 B
ATOM 3578 CD GLU B 263 56.461 32.869 23.413 1.00 20.70 B
ATOM 3579 OE1 GLU B 263 55.999 32.134 22.516 1.00 22.06 B
ATOM 3580 OE2 GLU B 263 55.754 33.670 24.063 1.00 22.68 B
ATOM 3581 C GLU B 263 60.548 29.919 24.101 1.00 12.75 B
ATOM 3582 0 GLU B 263 60.744 29.471 25.235 1.00 13.64 B
ATOM 3583 N ILE B 264 60.773 29.209 22.997 1.00 12.68 B
ATOM 3584 CA ILE B 264 61.295 27.850 23.063 1.00 12.55 B
ATOM 3585 CB ILE B 264 61.262 27.177 21.671 1.00 14.12 B
ATOM 3586 CG2 ILE B 264 62.131 25.935 21.659 1.00 14.08 B
ATOM 3587 CG1 ILE B 264 59.817 26.817 21.313 1.00 15.80 B
ATOM 3588 CD1 ILE B 264 59.634 26.352 19.881 1.00 20.14 B
ATOM 3589 C ILE B 264 62.730 27.921 23.582 1.00 12.33 B
ATOM 3590 0 ILE B 264 63.122 27.138 24.451 1.00 12.84 B
ATOM 3591 N LEU B 265 63.500 28.872 23.055 1.00 11.54 B
ATOM 3592 CA LEU B 265 64.887 29.073 23.477 1.00 12.06 B
ATOM 3593 CB LEU B 265 65.497 30.265 22.733 1.00 13.23 B
ATOM 3594 CG LEU B 265 66.881 30.718 23.203 1.00 13.80 B
ATOM 3595 CD1 LEU B 265 67.883 29.587 23.020 1.00 15.33 B
ATOM 3596 CD2 LEU B 265 67.312 31.950 22.431 1.00 13.99 B
ATOM 3597 C LEU B 265 64.933 29.342 24.981 1.00 12.16 B
ATOM 3598 0 LEU B 265 65.746 28.768 25.698 1.00 11.75 B
ATOM 3599 N ALA B 266 64.047 30.217 25.447 1.00 12.67 B
ATOM 3600 CA ALA B 266 63.984 30.573 26.858 1.00 11.14 B
ATOM 3601 CB ALA B 266 62.955 31.686 27.066 1.00 11.89 B
ATOM 3602 C ALA B 266 63.645 29.391 27.757 1.00 10.09 B
ATOM 3603 0 ALA B 266 64.267 29.205 28.802 1.00 11.77 B
ATOM 3604 N ALA B 267 62.653 28.599 27.357 1.00 10.44 B
ATOM 3605 CA ALA B 267 62.225 27.456 28.156 1.00 10.99 B
ATOM 3606 CB ALA B 267 60.982 26.817 27.533 1.00 12.27 B

ATOM 3607 C ALA B 267 63.320 26.410 28.313 1.00 11.14 B
ATOM 3608 0 ALA B 267 63.574 25.916 29.414 1.00 12.10 B
ATOM 3609 N ILE B 268 63.973 26.067 27.211 1.00 11.40 B
ATOM 3610 CA ILE B 268 65.028 25.067 27.269 1.00 12.32 B
ATOM 3611 CB ILE B 268 65.433 24.617 25.852 1.00 14.56 B
ATOM 3612 CG2 ILE B 268 66.489 23.528 25.931 1.00 16.64 B
ATOM 3613 CG1 ILE B 268 64.203 24.054 25.129 1.00 16.27 B
ATOM 3614 CD1 ILE B 268 64.454 23.659 23.680 1.00 19.23 B
ATOM 3615 C ILE B 268 66.234 25.596 28.044 1.00 11.25 B
ATOM 3616 0 ILE B 268 66.850 24.850 28.809 1.00 12.03 B
ATOM 3617 N PHE B 269 66.553 26.878 27.867 1.00 11.57 B
ATOM 3618 CA PHE B 269 67.672 27.492 28.587 1.00 10.70 B
ATOM 3619 CB PHE B 269 67.906 28.925 28.093 1.00 11.45 B
ATOM 3620 CG PHE B 269 68.996 29.658 28.835 1.00 12.99 B
ATOM 3621 CD1 PHE B 269 70.333 29.354 28.624 1.00 14.49 B
ATOM 3622 CD2 PHE B 269 68.677 30.653 29.744 1.00 13.23 B
ATOM 3623 CE1 PHE B 269 71.333 30.038 29.311 1.00 14.66 B
ATOM 3624 CE2 PHE B 269 69.666 31.337 30.434 1.00 13.64 B
ATOM 3625 CZ PHE B 269 70.995 31.029 30.215 1.00 13.99 B
ATOM 3626 C PHE B 269 67.345 27.512 30.080 1.00 10.43 B
ATOM 3627 0 PHE B 269 68.188 27.185 30.922 1.00 9.52 B
ATOM 3628 N ALA B 270 66.116 27.896 30.408 1.00 10.31 B
ATOM 3629 CA ALA B 270 65.703 27.934 31.804 1.00 9.54 B
ATOM 3630 CB ALA B 270 64.247 28.397 31.915 1.00 10.92 B
ATOM 3631 C ALA B 270 65.879 26.552 32.427 1.00 9.80 B
ATOM 3632 0 ALA B 270 66.402 26.425 33.530 1.00 11.73 B
ATOM 3633 N ALA B 271 65.446 25.509 31.721 1.00 10.57 B
ATOM 3634 CA ALA B 271 65.596 24.154 32.242 1.00 10.67 B
ATOM 3635 CB ALA B 271 64.967 23.149 31.268 1.00 13.41 B
ATOM 3636 C ALA B 271 67.072 23.805 32.482 1.00 9.22 B
ATOM 3637 0 ALA B 271 67.422 23.216 33.502 1.00 11.13 B
ATOM 3638 N ALA B 272 67.946 24.173 31.550 1.00 8.45 B
ATOM 3639 CA ALA B 272 69.368 23.865 31.694 1.00 8.29 B
ATOM 3640 CB ALA B 272 70.115 24.252 30.414 1.00 9.71 B
ATOM 3641 C ALA B 272 70.048 24.518 32.902 1.00 9.17 B
ATOM 3642 0 ALA B 272 70.889 23.904 33.544 1.00 11.32 B
ATOM 3643 N ILE B 273 69.676 25.755 33.214 1.00 9.92 B
ATOM 3644 CA ILE B 273 70.299 26.470 34.330 1.00 10.55 B
ATOM 3645 CB ILE B 273 70.510 27.967 34.007 1.00 11.17 B
ATOM 3646 CG2 ILE B 273 71.200 28.134 32.660 1.00 12.80 B
ATOM 3647 CG1 ILE B 273 69.160 28.688 34.000 1.00 10.79 B
ATOM 3648 CD1 ILE B 273 69.282 30.209 34.026 1.00 10.44 B
ATOM 3649 C ILE B 273 69.497 26.471 35.617 1.00 9.63 B
ATOM 3650 0 ILE B 273 69.980 26.958 36.636 1.00 11.91 B
ATOM 3651 N HIS B 274 68.292 25.917 35.591 1.00 10.09 B
ATOM 3652 CA HIS B 274 67.421 25.998 36.758 1.00 8.74 B
ATOM 3653 CB HIS B 274 66.060 25.356 36.447 1.00 9.43 B
ATOM 3654 CG HIS B 274 65.952 23.925 36.864 1.00 10.96 B
ATOM 3655 CD2 HIS B 274 65.392 23.360 37.960 1.00 8.81 B
ATOM 3656 ND1 HIS B 274 66.509 22.891 36.141 1.00 10.16 B
ATOM 3657 CE1 HIS B 274 66.300 21.752 36.777 1.00 10.39 B
ATOM 3658 NE2 HIS B 274 65.625 22.010 37.884 1.00 7.84 B
ATOM 3659 C HIS B 274 67.915 25.539 38.131 1.00 8.71 B
ATOM 3660 0 HIS B 274 67.319 25.929 39.147 1.00 9.75 B
ATOM 3661 N ASP B 275 68.972 24.725 38.182 1.00 8.42 B
ATOM 3662 CA ASP B 275 69.531 24.258 39.466 1.00 8.17 B
ATOM 3663 CB ASP B 275 69.221 22.769 39.711 1.00 8.97 B
ATOM 3664 CG ASP B 275 67.884 22.541 40.381 1.00 10.33 B
ATOM 3665 001 ASP B 275 67.447 23.412 41.148 1.00 10.14 B
ATOM 3666 OD2 ASP B 275 67.274 21.475 40.161 1.00 9.86 B
ATOM 3667 C ASP B 275 71.055 24.433 39.548 1.00 8.46 B
ATOM 3668 0 ASP B 275 71.708 23.825 40.404 1.00 9.88 B
ATOM 3669 N VAL B 276 71.630 25.255 38.675 1.00 9.20 B
ATOM 3670 CA VAL B 276 73.083 25.425 38.680 1.00 8.98 B
ATOM 3671 CB VAL B 276 73.541 26.351 37.523 1.00 10.25 B
ATOM 3672 CG1 VAL B 276 73.086 27.779 37.756 1.00 11.35 B
ATOM 3673 CG2 VAL B 276 75.045 26.265 37.367 1.00 11.42 B

ATOM 3674 C VAL B 276 73.672 25.907 40.011 1.00 8.42 B
ATOM 3675 0 VAL B 276 73.123 26.778 40.680 1.00 8.82 B
ATOM 3676 N ASP B 277 74.794 25.305 40.391 1.00 9.58 B
ATOM 3677 CA ASP B 277 75.500 25.624 41.634 1.00 10.27 B
ATOM 3678 CB ASP B 277 75.943 27.096 41.638 1.00 13.36 B
ATOM 3679 CG ASP B 277 77.058 27.368 42.637 1.00 16.91 B
ATOM 3680 OD1 ASP B 277 77.885 26.458 42.865 1.00 17.94 B
ATOM 3681 OD2 ASP B 277 77.123 28.494 43.181 1.00 17.97 B
ATOM 3682 C ASP B 277 74.672 25.298 42.884 1.00 10.70 B
ATOM 3683 0 ASP B 277 74.793 25.956 43.915 1.00 13.58 B
ATOM 3684 N HIS B 278 73.841 24.266 42.779 1.00 10.79 B
ATOM 3685 CA HIS B 278 73.005 23.806 43.887 1.00 10.24 B
ATOM 3686 CB HIS B 278 71.975 22.790 43.378 1.00 9.80 B
ATOM 3687 CG HIS B 278 70.826 22.562 44.313 1.00 10.59 B
ATOM 3688 CD2 HIS B 278 69.500 22.783 44.158 1.00 12.43 B
ATOM 3689 ND1 HIS B 278 70.981 22.031 45.577 1.00 13.40 B
ATOM 3690 CE1 HIS B 278 69.798 21.935 46.159 1.00 11.93 B
ATOM 3691 NE2 HIS B 278 68.882 22.384 45.319 1.00 12.85 B
ATOM 3692 C HIS B 278 73.946 23.133 44.880 1.00 9.88 B
ATOM 3693 0 HIS B 278 74.720 22.247 44.510 1.00 12.94 B
ATOM 3694 N PRO B 279 73.900 23.552 46.151 1.00 11.89 B
ATOM 3695 CD PRO B 279 73.131 24.699 46.664 1.00 12.25 B
ATOM 3696 CA PRO B 279 74.760 22.985 47.192 1.00 12.36 B
ATOM 3697 CB PRO B 279 74.794 24.094 48.239 1.00 13.43 B
ATOM 3698 CG PRO B 279 73.418 24.651 48.151 1.00 13.68 B
ATOM 3699 C PRO B 279 74.307 21.648 47.774 1.00 12.90 B
ATOM 3700 0 PRO B 279 75.002 21.076 48.612 1.00 14.71 B
ATOM 3701 N GLY B 280 73.155 21.151 47.333 1.00 12.90 B
ATOM 3702 CA GLY B 280 72.670 19.876 47.837 1.00 13.10 B
ATOM 3703 C GLY B 280 71.915 19.957 49.150 1.00 13.99 B
ATOM 3704 0 GLY B 280 71.732 18.942 49.824 1.00 14.55 B
ATOM 3705 N VAL B 281 71.500 21.164 49.523 1.00 14.58 B
ATOM 3706 CA VAL B 281 70.721 21.380 50.741 1.00 14.37 B
ATOM 3707 CB VAL B 281 71.567 22.041 51.861 1.00 15.47 B
ATOM 3708 CG1 VAL B 281 72.657 21.084 52.319 1.00 16.54 B
ATOM 3709 CG2 VAL B 281 72.182 23.336 51.363 1.00 17.92 B
ATOM 3710 C VAL B 281 69.533 22.277 50.392 1.00 14.06 B
ATOM 3711 0 VAL B 281 69.571 23.022 49.409 1.00 15.98 B
ATOM 3712 N SER B 282 68.480 22.204 51.199 1.00 12.90 B
ATOM 3713 CA SER B 282 67.261 22.979 50.972 1.00 12.27 B
ATOM 3714 CB SER B 282 66.129 22.405 51.818 1.00 15.81 B
ATOM 3715 OG SER B 282 66.389 22.624 53.197 1.00 16.87 B
ATOM 3716 C SER B 282 67.393 24.463 51.296 1.00 11.54 B
ATOM 3717 0 SER B 282 68.340 24.886 51.958 1.00 13.10 B
ATOM 3718 N ASN B 283 66.427 25.252 50.832 1.00 12.56 B
ATOM 3719 CA ASN B 283 66.432 26.681 51.111 1.00 10.95 B
ATOM 3720 CB ASN B 283 65.208 27.381 50.504 1.00 12.93 B
ATOM 3721 CG ASN B 283 65.400 27.741 49.044 1.00 13.52 B
ATOM 3722 OD1 ASN B 283 66.492 28.116 48.626 1.00 17.03 B
ATOM 3723 ND2 ASN B 283 64.327 27.655 48.267 1.00 14.50 B
ATOM 3724 C ASN B 283 66.391 26.863 52.623 1.00 11.55 B
ATOM 3725 0 ASN B 283 67.085 27.709 53.174 1.00 12.29 B
ATOM 3726 N GLN B 284 65.577 26.054 53.292 1.00 12.65 B
ATOM 3727 CA GLN B 284 65.453 26.178 54.740 1.00 13.65 B
ATOM 3728 CB GLN B 284 64.423 25.188 55.284 1.00 16.47 B
ATOM 3729 CG GLN B 284 63.922 25.560 56.675 1.00 22.61 B
ATOM 3730 CD GLN B 284 63.376 26.980 56.733 1.00 23.14 B
ATOM 3731 OE1 GLN B 284 62.491 27.351 55.961 1.00 25.00 B
ATOM 3732 NE2 GLN B 284 63.905 27.780 57.649 1.00 23.80 B
ATOM 3733 C GLN B 284 66.794 25.972 55.439 1.00 12.32 B
ATOM 3734 0 GLN B 284 67.103 26.654 56.409 1.00 13.35 B
ATOM 3735 N PHE B 285 67.588 25.030 54.945 1.00 12.52 B
ATOM 3736 CA PHE B 285 68.902 24.781 55.537 1.00 11.32 B
ATOM 3737 CB PHE B 285 69.572 23.593 54.829 1.00 11.69 B
ATOM 3738 CG PHE B 285 70.931 23.237 55.371 1.00 12.84 B
ATOM 3739 CD1 PHE B 285 72.053 23.978 55.026 1.00 15.56 B
ATOM 3740 CD2 PHE B 285 71.094 22.134 56.195 1.00 14.79 B

ATOM 3741 CE1 PHE B 285 73.310 23.623 55.487 1.00 14.41 B
ATOM 3742 CE2 PHE B 285 72.352 21.776 56.660 1.00 12.95 B
ATOM 3743 CZ PHE B 285 73.456 22.519 56.304 1.00 14.80 B
ATOM 3744 C PHE B 285 69.765 26.041 55.407 1.00 11.25 B
ATOM 3745 0 PHE B 285 70.441 26.443 56.351 1.00 12.39 B
ATOM 3746 N LEU B 286 69.745 26.659 54.230 1.00 11.60 B
ATOM 3747 CA LEU B 286 70.527 27.868 53.995 1.00 11.39 B
ATOM 3748 CB LEU B 286 70.412 28.289 52.531 1.00 13.01 B
ATOM 3749 CG LEU B 286 71.005 27.298 51.530 1.00 15.88 B
ATOM 3750 CD1 LEU B 286 70.748 27.802 50.118 1.00 16.02 B
ATOM 3751 CD2 LEU B 286 72.495 27.136 51.775 1.00 16.17 B
ATOM 3752 C LEU B 286 70.056 28.999 54.903 1.00 11.62 B
ATOM 3753 0 LEU B 286 70.861 29.754 55.450 1.00 13.36 B
ATOM 3754 N ILE B 287 68.746 29.114 55.067 1.00 12.47 B
ATOM 3755 CA ILE B 287 68.195 30.156 55.919 1.00 13.31 B
ATOM 3756 CB ILE B 287 66.659 30.190 55.819 1.00 14.28 B
ATOM 3757 CG2 ILE B 287 66.090 31.169 56.843 1.00 13.43 B
ATOM 3758 CG1 ILE B 287 66.242 30.594 54.405 1.00 15.88 B
ATOM 3759 CD1 ILE B 287 64.766 30.403 54.119 1.00 15.36 B
ATOM 3760 C ILE B 287 68.598 29.909 57.377 1.00 12.47 B
ATOM 3761 0 ILE B 287 69.023 30.833 58.073 1.00 14.12 B
ATOM 3762 N ASN B 288 68.488 28.662 57.825 1.00 13.80 B
ATOM 3763 CA ASN B 288 68.821 28.314 59.210 1.00 15.32 B
ATOM 3764 CB ASN B 288 68.394 26.877 59.523 1.00 16.95 B
ATOM 3765 CG ASN B 288 66.888 26.709 59.561 1.00 19.11 B
ATOM 3766 OD1 ASN B 288 66.149 27.676 59.739 1.00 21.37 B
ATOM 3767 ND2 ASN B 288 66.426 25.472 59.411 1.00 22.68 B
ATOM 3768 C ASN B 288 70.296 28.476 59.568 1.00 15.24 B
ATOM 3769 0 ASN B 288 70.634 28.699 60.734 1.00 18.51 B
ATOM 3770 N THR B 289 71.172 28.353 58.577 1.00 14.49 B
ATOM 3771 CA THR B 289 72.603 28.490 58.815 1.00 14.96 B
ATOM 3772 CB THR B 289 73.416 27.516 57.923 1.00 14.70 B
ATOM 3773 001 THR B 289 73.073 27.714 56.546 1.00 14.76 B
ATOM 3774 CG2 THR B 289 73.123 26.082 58.309 1.00 16.87 B
ATOM 3775 C THR B 289 73.097 29.914 58.587 1.00 14.30 B
ATOM 3776 0 THR B 289 74.296 30.186 58.675 1.00 16.29 B
ATOM 3777 N ASN B 290 72.170 30.823 58.295 1.00 15.22 B
ATOM 3778 CA ASN B 290 72.508 32.222 58.058 1.00 16.21 B
ATOM 3779 CB ASN B 290 73.103 32.840 59.327 1.00 18.93 B
ATOM 3780 CG ASN B 290 73.337 34.334 59.200 1.00 23.38 B
ATOM 3781 OD1 ASN B 290 74.347 34.860 59.679 1.00 26.84 B
ATOM 3782 ND2 ASN B 290 72.398 35.028 58.567 1.00 23.94 B
ATOM 3783 C ASN B 290 73.520 32.319 56.924 1.00 15.21 B
ATOM 3784 0 ASN B 290 74.451 33.122 56.972 1.00 16.68 B
ATOM 3785 N SER B 291 73.342 31.492 55.903 1.00 15.31 B
ATOM 3786 CA SER B 291 74.258 31.503 54.771 1.00 16.41 B
ATOM 3787 CB SER B 291 73.915 30.368 53.811 1.00 18.28 B
ATOM 3788 OG SER B 291 72.647 30.585 53.228 1.00 23.19 B
ATOM 3789 C SER B 291 74.217 32.830 54.017 1.00 14.89 B
ATOM 3790 0 SER B 291 73.197 33.522 54.001 1.00 15.57 B
ATOM 3791 N GLU B 292 75.339 33.174 53.391 1.00 16.31 B
ATOM 3792 CA GLU B 292 75.457 34.404 52.614 1.00 17.02 B
ATOM 3793 CB GLU B 292 76.840 34.471 51.962 1.00 19.06 B
ATOM 3794 CG GLU B 292 77.040 35.642 51.018 1.00 23.64 B
ATOM 3795 CD GLU B 292 78.446 35.690 50.450 1.00 26.32 B
ATOM 3796 OE1 GLU B 292 78.893 34.671 49.876 1.00 28.58 B
ATOM 3797 OE2 GLU B 292 79.104 36.745 50.579 1.00 29.58 B
ATOM 3798 C GLU B 292 74.373 34.506 51.546 1.00 16.12 B
ATOM 3799 0 GLU B 292 73.885 35.597 51.259 1.00 17.12 B
ATOM 3800 N LEU B 293 73.985 33.374 50.963 1.00 16.26 B
ATOM 3801 CA LEU B 293 72.944 33.383 49.932 1.00 15.17 B
ATOM 3802 CB LEU B 293 72.841 32.020 49.244 1.00 17.78 B
ATOM 3803 CG LEU B 293 73.959 31.654 48.272 1.00 19.61 B
ATOM 3804 CD1 LEU B 293 73.661 30.292 47.664 1.00 22.08 B
ATOM 3805 CD2 LEU B 293 74.066 32.721 47.190 1.00 23.02 B
ATOM 3806 C LEU B 293 71.580 33.749 50.500 1.00 15.21 B
ATOM 3807 0 LEU B 293 70.816 34.493 49.876 1.00 15.20 B

ATOM 3808 N ALA B 294 71.269 33.222 51.681 1.00 13.95 B
ATOM 3809 CA ALA B 294 69.987 33.518 52.311 1.00 14.53 B
ATOM 3810 CB ALA B 294 69.800 32.674 53.574 1.00 14.29 B
ATOM 3811 C ALA B 294 69.914 34.999 52.649 1.00 15.22 B
ATOM 3812 0 ALA B 294 68.865 35.621 52.509 1.00 16.21 B
ATOM 3813 N LEU B 295 71.029 35.571 53.096 1.00 15.84 B
ATOM 3814 CA LEU B 295 71.045 36.993 53.421 1.00 16.78 B
ATOM 3815 CB LEU B 295 72.331 37.363 54.164 1.00 17.35 B
ATOM 3816 CG LEU B 295 72.378 36.975 55.640 1.00 19.95 B
ATOM 3817 CD1 LEU B 295 73.696 37.422 56.239 1.00 23.50 B
ATOM 3818 CD2 LEU B 295 71.220 37.627 56.369 1.00 22.71 B
ATOM 3819 C LEU B 295 70.943 37.809 52.140 1.00 16.38 B
ATOM 3820 0 LEU B 295 70.273 38.841 52.092 1.00 18.26 B
ATOM 3821 N MET B 296 71.601 37.329 51.094 1.00 16.17 B
ATOM 3822 CA MET B 296 71.591 38.012 49.809 1.00 16.75 B
ATOM 3823 CB MET B 296 72.461 37.243 48.811 1.00 19.70 B
ATOM 3824 CG MET B 296 73.648 38.026 48.285 1.00 28.48 B
ATOM 3825 SD MET B 296 73.137 39.457 47.313 1.00 24.77 B
ATOM 3826 CE MET B 296 73.324 38.831 45.628 1.00 30.08 B
ATOM 3827 C MET B 296 70.190 38.172 49.231 1.00 15.03 B
ATOM 3828 0 MET B 296 69.848 39.229 48.693 1.00 16.71 B
ATOM 3829 N TYR B 297 69.378 37.126 49.353 1.00 14.35 B
ATOM 3830 CA TYR B 297 68.031 37.141 48.799 1.00 12.40 B
ATOM 3831 CB TYR B 297 67.868 35.926 47.876 1.00 12.95 B
ATOM 3832 CG TYR B 297 68.806 36.003 46.694 1.00 12.48 B
ATOM 3833 CD1 TYR B 297 68.680 37.024 45.762 1.00 13.15 B
ATOM 3834 CE1 TYR B 297 69.573 37.158 44.715 1.00 12.70 B
ATOM 3835 CD2 TYR B 297 69.859 35.104 46.543 1.00 12.71 B
ATOM 3836 CE2 TYR B 297 70.766 35.234 45.493 1.00 13.85 B
ATOM 3837 CZ TYR B 297 70.613 36.267 44.585 1.00 12.18 B
ATOM 3838 OH TYR B 297 71.509 36.432 43.548 1.00 15.45 B
ATOM 3839 C TYR B 297 66.885 37.228 49.812 1.00 12.43 B
ATOM 3840 0 TYR B 297 65.746 36.876 49.510 1.00 13.42 B
ATOM 3841 N ASN B 298 67.196 37.719 51.008 1.00 13.10 B
ATOM 3842 CA ASN B 298 66.192 37.891 52.059 1.00 14.27 B
ATOM 3843 CB ASN B 298 65.269 39.064 51.706 1.00 15.38 B
ATOM 3844 CG ASN B 298 66.036 40.317 51.330 1.00 18.06 B
ATOM 3845 OD1 ASN B 298 66.935 40.754 52.052 1.00 21.15 B
ATOM 3846 ND2 ASN B 298 65.675 40.911 50.196 1.00 20.44 B
ATOM 3847 C ASN B 298 65.335 36.658 52.363 1.00 15.06 B
ATOM 3848 0 ASN B 298 64.132 36.775 52.614 1.00 17.05 B
ATOM 3849 N ASP B 299 65.961 35.486 52.345 1.00 15.30 B
ATOM 3850 CA ASP B 299 65.286 34.219 52.627 1.00 16.00 B
ATOM 3851 CB ASP B 299 64.708 34.211 54.051 1.00 17.15 B
ATOM 3852 CG ASP B 299 65.753 34.464 55.124 1.00 20.00 B
ATOM 3853 OD1 ASP B 299 66.954 34.219 54.886 1.00 21.27 B
ATOM 3854 OD2 ASP B 299 65.359 34.895 56.234 1.00 23.31 B
ATOM 3855 C ASP B 299 64.162 33.844 51.661 1.00 15.85 B
ATOM 3856 0 ASP B 299 63.413 32.904 51.927 1.00 17.12 B
ATOM 3857 N GLU B 300 64.043 34.561 50.544 1.00 15.59 B
ATOM 3858 CA GLU B 300 62.987 34.287 49.568 1.00 17.14 B
ATOM 3859 CB GLU B 300 62.287 35.592 49.189 1.00 19.40 B
ATOM 3860 CG GLU B 300 61.459 36.203 50.307 1.00 24.35 B
ATOM 3861 CD GLU B 300 60.060 35.624 50.381 1.00 28.50 B
ATOM 3862 OE1 GLU B 300 59.924 34.396 50.572 1.00 30.02 B
ATOM 3863 OE2 GLU B 300 59.091 36.403 50.242 1.00 31.25 B
ATOM 3864 C GLU B 300 63.538 33.614 48.316 1.00 16.27 B
ATOM 3865 0 GLU B 300 64.393 34.178 47.638 1.00 17.16 B
ATOM 3866 N SER B 301 63.030 32.419 48.010 1.00 15.30 B
ATOM 3867 CA SER B 301 63.492 31.647 46.854 1.00 14.03 B
ATOM 3868 CB SER B 301 62.819 32.138 45.573 1.00 14.54 B
ATOM 3869 OG SER B 301 61.420 31.942 45.648 1.00 17.57 B
ATOM 3870 C SER B 301 64.999 31.799 46.733 1.00 11.76 B
ATOM 3871 0 SER B 301 65.521 32.126 45.671 1.00 13.15 B
ATOM 3872 N VAL B 302 65.696 31.541 47.835 1.00 12.29 B
ATOM 3873 CA VAL B 302 67.142 31.694 47.886 1.00 11.28 B
ATOM 3874 CB VAL B 302 67.683 31.263 49.260 1.00 12.31 B

ATOM 3875 CG1 VAL B 302 69.182 31.475 49.318 1.00 12.16 B
ATOM 3876 CG2 VAL B 302 66.992 32.064 50.358 1.00 13.10 B
ATOM 3877 C VAL B 302 67.899 30.946 46.792 1.00 11.14 B
ATOM 3878 0 VAL B 302 68.670 31.549 46.045 1.00 12.80 B
ATOM 3879 N LEU B 303 67.681 29.639 46.700 1.00 11.29 B
ATOM 3880 CA LEU B 303 68.355 28.820 45.695 1.00 12.12 B
ATOM 3881 CB LEU B 303 67.984 27.344 45.879 1.00 14.58 B
ATOM 3882 CG LEU B 303 68.611 26.624 47.076 1.00 17.98 B
ATOM 3883 CD1 LEU B 303 67.884 25.308 47.324 1.00 19.99 B
ATOM 3884 CD2 LEU B 303 70.080 26.392 46.804 1.00 19.30 B
ATOM 3885 C LEU B 303 68.000 29.242 44.274 1.00 10.31 B
ATOM 3886 0 LEU B 303 68.876 29.414 43.429 1.00 11.10 B
ATOM 3887 N GLU B 304 66.708 29.396 44.014 1.00 10.04 B
ATOM 3888 CA GLU B 304 66.250 29.775 42.687 1.00 9.51 B
ATOM 3889 CB GLU B 304 64.723 29.770 42.671 1.00 11.48 B
ATOM 3890 CG GLU B 304 64.110 28.379 42.943 1.00 12.15 B
ATOM 3891 CD GLU B 304 64.255 27.885 44.387 1.00 14.37 B
ATOM 3892 OE1 GLU B 304 64.325 28.719 45.318 1.00 14.49 B
ATOM 3893 OE2 GLU B 304 64.271 26.646 44.591 1.00 15.25 B
ATOM 3894 C GLU B 304 66.823 31.119 42.216 1.00 9.19 B
ATOM 3895 0 GLU B 304 67.180 31.270 41.049 1.00 12.10 B
ATOM 3896 N ASN B 305 66.919 32.098 43.115 1.00 10.01 B
ATOM 3897 CA ASN B 305 67.508 33.381 42.737 1.00 10.32 B
ATOM 3898 CB ASN B 305 67.394 34.386 43.880 1.00 11.91 B
ATOM 3899 CG ASN B 305 66.110 35.184 43.827 1.00 12.96 B
ATOM 3900 OD1 ASN B 305 65.898 35.976 42.911 1.00 14.58 B
ATOM 3901 ND2 ASN B 305 65.243 34.976 44.810 1.00 14.47 B
ATOM 3902 C ASN B 305 68.986 33.186 42.391 1.00 10.29 B
ATOM 3903 0 ASN B 305 69.514 33.820 41.474 1.00 11.48 B
ATOM 3904 N HIS B 306 69.649 32.309 43.135 1.00 8.94 B
ATOM 3905 CA HIS B 306 71.057 32.027 42.897 1.00 9.84 B
ATOM 3906 CB HIS B 306 71.619 31.185 44.045 1.00 10.55 B
ATOM 3907 CG HIS B 306 73.086 30.927 43.933 1.00 13.66 B
ATOM 3908 CD2 HIS B 306 73.785 29.771 43.854 1.00 15.08 B
ATOM 3909 ND1 HIS B 306 74.015 31.943 43.881 1.00 15.44 B
ATOM 3910 CE1 HIS B 306 75.224 31.423 43.775 1.00 16.32 B
ATOM 3911 NE2 HIS B 306 75.112 30.106 43.756 1.00 14.88 B
ATOM 3912 C HIS B 306 71.293 31.315 41.562 1.00 8.93 B
ATOM 3913 0 HIS B 306 72.241 31.636 40.842 1.00 10.07 B
ATOM 3914 N HIS B 307 70.436 30.351 41.228 1.00 7.61 B
ATOM 3915 CA HIS B 307 70.598 29.623 39.972 1.00 7.78 B
ATOM 3916 CB HIS B 307 69.495 28.569 39.811 1.00 7.06 B
ATOM 3917 CG HIS B 307 69.414 27.598 40.950 1.00 7.95 B
ATOM 3918 CD2 HIS B 307 68.347 27.046 41.577 1.00 8.54 B
ATOM 3919 ND1 HIS B 307 70.532 27.057 41.549 1.00 9.97 B
ATOM 3920 CE1 HIS B 307 70.157 26.214 42.495 1.00 9.57 B
ATOM 3921 NE2 HIS B 307 68.837 26.187 42.531 1.00 9.41 B
ATOM 3922 C HIS B 307 70.540 30.619 38.818 1.00 6.60 B
ATOM 3923 0 HIS B 307 71.338 30.551 37.878 1.00 10.13 B
ATOM 3924 N LEU B 308 69.584 31.539 38.895 1.00 8.24 B
ATOM 3925 CA LEU B 308 69.421 32.566 37.875 1.00 9.36 B
ATOM 3926 CB LEU B 308 68.185 33.418 38.177 1.00 11.02 B
ATOM 3927 CG LEU B 308 66.838 32.790 37.804 1.00 11.82 B
ATOM 3928 CD1 LEU B 308 65.699 33.580 38.437 1.00 11.51 B
ATOM 3929 CD2 LEU B 308 66.694 32.766 36.287 1.00 11.00 B
ATOM 3930 C LEU B 308 70.651 33.462 37.794 1.00 10.02 B
ATOM 3931 0 LEU B 308 71.168 33.722 36.711 1.00 11.77 B
ATOM 3932 N ALA B 309 71.128 33.932 38.941 1.00 10.08 B
ATOM 3933 CA ALA B 309 72.302 34.801 38.953 1.00 9.46 B
ATOM 3934 CB ALA B 309 72.626 35.212 40.386 1.00 13.55 B
ATOM 3935 C ALA B 309 73.518 34.121 38.315 1.00 9.19 B
ATOM 3936 0 ALA B 309 74.223 34.722 37.506 1.00 11.81 B
ATOM 3937 N VAL B 310 73.769 32.874 38.691 1.00 8.85 B
ATOM 3938 CA VAL B 310 74.893 32.126 38.146 1.00 9.84 B
ATOM 3939 CB VAL B 310 75.058 30.783 38.874 1.00 9.88 B
ATOM 3940 CG1 VAL B 310 76.119 29.939 38.182 1.00 12.31 B
ATOM 3941 CG2 VAL B 310 75.447 31.034 40.328 1.00 9.44 B

DEMANDE OU BREVET VOLUMINEUX

LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET COMPREND
PLUS D'UN TOME.

NOTE : Pour les tomes additionels, veuillez contacter le Bureau canadien des brevets JUMBO APPLICATIONS/PATENTS

THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE THAN ONE
VOLUME

NOTE: For additional volumes, please contact the Canadian Patent Office NOM DU FICHIER / FILE NAME:

NOTE POUR LE TOME / VOLUME NOTE:

Claims (99)

1. A compound having the chemical structure all salts, prodrugs, tautomers and isomers thereof, wherein:
k is selected from the group consisting of -CR6R7R19, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14;

Z is O, S, or NR9;
t is CH;
y is N or CH;
v, w and x are CH;
A has a structure selected from the group consisting of R6 and R7 are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylakyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R8 at each occurrence is independently selected from the group consisting of -OR9, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R8 is alkenyl, no alkene carbon thereof is bound to C(Z), optionally substituted lower alkynyl, provided, however, that when R8 is alkynyl, no alkyne carbon thereof is bound to C(Z), optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R9 at each occurrence is independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R9 is alkenyl, no alkene carbon thereof is bound to O, N or S, optionally substituted lower alkynyl, provided, however, that when R9 is alkynyl, no alkyne carbon thereof is bound to O, N or S, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R10 and R11 at each occurrence are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R10 and/or R11 are alkenyl, no alkene carbon thereof is bound to nitrogen, optionally substituted lower alkynyl, provided, however, that when R10 and/or R11 are alkynyl, no alkyne carbon thereof is bound to nitrogen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14; or R10 and R11 together with the nitrogen to which they are attached form a 5-7 membered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring;
R12 and R13 at each occurrence are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R12 and/or R13 are alkenyl, no alkene carbon thereof is bound to nitrogen, optionally substituted lower alkynyl, provided, however, that when R12 and/or R13 are alkynyl, no alkyne carbon thereof is bound to nitrogen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl; or R12 and R13 together with the nitrogen to which they are attached form a 5-7 membered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring;
R14 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R14 is alkenyl, no alkene carbon thereof is bound to -S(O)2-, optionally substituted lower alkynyl, provided, however, that when R14 is alkynyl, no alkyne carbon thereof is bound to -S(O)2-, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted hoteroaralkyl;
R15 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R15 is alkenyl, no alkene carbon thereof is bound to oxygen, optionally substituted lower alkynyl, provided, however, that when R15 is alkynyl, no alkyne carbon thereof is bound to oxygen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, and -C(Z)NR12R13; or each R15, along with the oxygens to which they are bound, combine to form a 5-membered optionally substituted heterocycloalkyl ring fused to the phenyl ring;
R16 is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl provided, however, that when R16 is alkenyl, no alkene carbon thereof is bound to oxygen, optionally substituted lower alkynyl provided, however, that when R16 is alkynyl, no alkyne carbon thereof is bound to oxygen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, and -C(Z)NR12R13;
R17 is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl provided, however, that when R17 is alkenyl, no alkene carbon thereof is bound to N, O, or S, optionally substituted lower alkynyl, provided, however, that when R17 is alkynyl, no alkyne carbon thereof is bound to N, O, or S, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R18 is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, -OR4, -SR9, -NR10R11, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14, and R19 is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
provided, however that the compound is not

2. The compound of claim 1, wherein y is CH.

3. The compound of claim 1, wherein y is N.

4. The compound of either of claims 3 or 99, wherein k is selected from the group consisting of -CH2R19, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14, and wherein R8, R12, R13, R14, and R19 are selected from the group consisting of optionally substituted lower alkyl, aryl and heteroaryl, wherein aryl and heteroaryl are optionally substituted with 1-3 substituents selected from the group consisting of halogen, optionally substituted lower alkyl, optionally substituted lower alkoxy, optionally substituted lower thioalkyl, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted amino, optionally substituted carboxyl, optionally substituted alkylsulfonylamino, cyano and nitro.

5. The compound of claim 3 or 99, wherein A has the structure

6. The compound of claim 5 having the structure

7. The compound of claim 5 having the structure

8. The compound of claim 5 having the structure

9. The compound of claim 8, wherein k is selected from the group consisting of -CH2R19, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14; and wherein R8, R12, R13, R14, and R19 are selected from the group consisting of optionally substituted lower alkyl, aryl and heteroaryl, wherein aryl and heteroaryl are optionally substituted with 1-3 substituents selected from the group consisting of halogen, optionally substituted lower alkyl, optionally substituted lower alkoxy, optionally substituted lower thioalkyl, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted amino, optionally substituted carboxyl, optionally substituted alkylsulfonylamino, cyano and nitro.

10. The compound of claim 9, wherein each occurrence of R15 is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl, or each R15, along with the oxygens to which they are bound, combine to form a 5-7 membered optionally substituted heterocycloalkyl ring fused to the phenyl ring.

11. The compound of claim 10, wherein when R15 is optionally substituted lower alkyl, the lower alkyl is optionally substituted with 1-3 substituents selected from the group consisting of fluoro, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl.

12. A composition comprising:
a compound according to any of claims 1-11, 96-98 or 99; and a pharmaceutically acceptable carrier.

13. A method for treating a subject suffering from or at risk of a disease or condition for which PDE4Y3 modulation provides a therapeutic benefit, comprising:
administering to said subject an effective amount of a compound having a chemical structure of wherein:
k is selected from the group consisting of -CR6R7R19, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14;
Z is O, S, or NR9;
t, u, v, w, x, and y are each independently N or CR1, provided, however, that no more than 1 of u and t are N, and no more than 2 of v, w, x and y are N;
R1 at each occurrence is independently selected from the group consisting of hydrogen, halogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylakyl, optionally substituted aralkyl, optionally substituted heteroaralkyl, -C(Z)R8, -OR9, -SR9, NR10R11, -C(Z)NR12R13, -S(O)2NR12R13, -S(O)2R14, and A, provided, however, that at least one R1 is A;
A is selected from the group consisting of substituted aryl and substituted heteroaryl;
R6 and R7 are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylakyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R8 at each occurrence is independently selected from the group consisting of -OR9, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R8 is alkenyl, no alkene carbon thereof is bound to C(Z), optionally substituted lower alkynyl, provided, however, that when R8 is alkynyl, no alkyne carbon thereof is bound to C(Z), optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R9 at each occurrence is independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R9 is alkenyl, no alkene carbon thereof is bound to O, N or S, optionally substituted lower alkynyl, provided, however, that when R9 is alkynyl, no alkyne carbon thereof is bound to O, N or S, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R10 and R11 are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R10 and/or R11 are alkenyl, no alkene carbon thereof is bound to nitrogen, optionally substituted lower alkynyl, provided, however, that when R10 and/or R11 are alkynyl, no alkyne carbon thereof is bound to nitrogen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14; or R10 and R11 together with the nitrogen to which they are attached form a 5-7 membered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring;
R12 and R13 at each occurrence are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R12 and/or R13 are alkenyl, no alkene carbon thereof is bound to nitrogen, optionally substituted lower alkynyl, provided, however, that when R12 and/or R13 are alkynyl, no alkyne carbon thereof is bound to nitrogen optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl; or R12 and R13 together with the nitrogen to which they are attached form a 5-7 membered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring;

R14 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R14 is alkenyl, no alkene carbon thereof is bound to -S(O)2-, optionally substituted lower alkynyl, provided, however, that when R14 is alkynyl, no alkyne carbon thereof is bound to -S(O)2-, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl; and R19 is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl.

14. The method of claim 13, wherein A has a structure selected from the group consisting of wherein;
R15 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R15 is alkenyl, no alkene carbon thereof is bound to oxygen, optionally substituted lower alkynyl, provided, however, that when R15 is alkynyl, no alkyne carbon thereof is bound to oxygen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted, heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, and -C(Z)NR12R13; or each R1s, along with the oxygens to which they are bound, combine to form a 5-membered optionally substituted heterocycloalkyl ring fused to the phenyl ring;
R16 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl provided, however; that when R16 is alkenyl, no alkene carbon thereof is bound to oxygen, optionally substituted lower alkynyl provided, however, that when R16 is alkynyl, no alkyne carbon thereof is bound to oxygen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R6, and -C(Z)NR12R13;

R17 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl provided, however, that when R17 is alkenyl, no alkene carbon thereof is bound to N, O, or S, optionally substituted lower alkynyl, provided, however, that when R17 is alkynyl, no alkyne carbon thereof is bound to N, O, or S, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl; and R18 is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, -OR9, -SR9, -NR10R11, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14.

15. The method of claim 14, wherein A has the structure

16. The method of claim 15, wherein no more than one of t, u, v, w, x, and y is N.

17. The method of claim 16, wherein t is CH, y is N, one of u, v, w, and x is C-A

and the others of u, v, w and x are CH.

18. The method of claim 17, wherein the compound has the structure

19. The method of claim 13, wherein said compound is approved for administration to a human.

20. The method of claim 19, wherein said disease or condition is a PDE4B-mediated disease or condition.

21. The method of claim 19, wherein said disease or condition is selected from the group consisting of asthma, bronchitis, allergic bronchitis, chronic obstructive pulmonary disease, cystic fibrosis, idiopathic pulmonary fibrosis, sarcoidosis, pulmonary hypertension, allergic bronchitis, emphysema, Alzheimer's disease, Parkinson's disease, Huntington's chorea, multiple sclerosis, rheumatoid arthritis, Crohn's disease, cerebral ischemia, inflammatory bowel disease, ulcerative colitis, atopic dermatitis, osteoporosis, osteopetrosis, Paget's disease, diffuse large-cell B cell lymphoma, chronic lymphocytic leukemia, acute lymphoblastic leukemia, Severe Acute Respiratory Syndrome, and pre-term labor.

22. A kit comprising a composition of claim 12.

23. The kit of claim 22, further comprising a written indication that said composition is approved for administering to a human.

24. The kit of claim 23, wherein said composition is approved for an indication selected from the group consisting of asthma, bronchitis, allergic bronchitis, chronic obstructive pulmonary disease, cystic fibrosis, idiopathic pulmonary fibrosis, sarcoidosis, pulmonary hypertension, allergic bronchitis, emphysema, Alzheimer's disease, Parkinson's disease, Huntington's chorea, multiple sclerosis, rheumatoid arthritis, Crohn's disease, cerebral ischemia, inflammatory bowel disease, ulcerative colitis, atopic dermatitis, osteoporosis, osteopetrosis, Paget's disease, diffuse large-cell B cell lymphoma, chronic lymphocytic leukemia, acute lymphoblastic leukemia, Severe Acute Respiratory Syndrome, and pre-term labor.

25. A method for developing an improved modulator active on PDE4, comprising:
determining whether any of a plurality of test compounds according to Formula I provides an improvement in one or more desired pharmacologic properties relative to a reference compound active on PDE4; and selecting those compounds that have an improvement in said desired pharmacologic property, thereby providing an improved modulator.

26. The method of claim 25, wherein said desired pharmacologic property is at least 10-fold greater activity on PDE4B than on PDE4D.

27. The method of claim 25, wherein said desired pharmacologic property is an of less than 0.10 µM.

28. The method of claim 25, wherein said reference compound is a compound of Formula I.

29. The method of claim 25, wherein at least one derivative of said improved modulator is used as a test compound and said determining and selecting are repeated,

30. A method for developing ligands with improved specificity for PDE4B, comprising:
identifying a compound that binds to a plurality of phosphodiesterases; and determining whether a derivative of said compound has greater specificity for PDE4B than said compound, thereby providing ligands with improved specificity for PDE4B.

31. The method of claim 30, wherein said compound binds to PDE4B with an affinity at least 10-fold greater than for binding to any of said plurality of phosphodiesterases.

32. The method of claim 30, wherein said compound interacts with at least one conserved PDE4B active site residue.

33. The method of claim 30, wherein said compound binds weakly to said plurality of phosphodiesterases.

34. The method of claim 30, wherein said plurality of phosphodiesterases comprises PDE4B and PDE4D.

35. The method of claim 30, wherein said plurality of phosphodiesterases comprises PDE4B and PDE5A.

36. The method of claim 30, wherein said compound is a compound of Formula I.

37. A crystal comprising a crystalline form of PDE4B phosphodiesterase domain, having coordinates as described in Table 1 or 2.

38. The crystal of claim 37, comprising one more heavy metal atoms.

39, The crystal of claim 37, wherein said crystalline form comprises a co-crystal of PDE4B with a binding compound.

40. The crystalline form of claim 39, wherein said binding compound interacts with at least one conserved PDE4B active site residue.

41. The crystalline form of claim 39, wherein said co-crystal is in an X-ray beam.

42. The crystalline form of claim, 37, wherein said crystal is in an X-ray beam.

43. A method for obtaining a crystal of PDE4B, comprising:
subjecting PDE4B protein at 5-20 mg/ml to crystallization condition substantially equivalent to 30% PEG 400, 0.2M MgCl2, 0.1M Tris pH 8.5, 1 mM binding compound, at 4 °C; or 20% PEG 3000, 0.2M Ca(OAc)2, 0.1M Tris pH 7.0, 1 mM binding compound, 15.9 mg/ml protein at 4 °C; or 1.8M-2.0M ammonium sulphate, 0.1 M
CAPS pH 10.0 - 10.5, 0.2M lithium sulphate.

44. The method of claim 43, further comprising optimizing said crystallization conditions.

45. A co-crystal of PDE4B and a PDE4B binding compound, wherein said binding compound comprises a compound of Formula I.

46. The co-crystal of claim 45, wherein said co-crystal is in an X-ray beam.

47. A method for determining a structure of a phosphodiesterase, comprising:
creating a homology model from an electronic representation of a PDE4B
structure, wherein said PDE4B structure represents the atomic coordinates of Table 1 or 2; and equating said homology model with said stucture of a phosphodiesterase.

48. The method of claim 47, wherein said creating comprises:
identifying conserved amino acid residues between PDE4B and said phosphodiesterase;
transferring the atomic coordinates of a plurality of conserved amino acids in said PDE4B
structure to the corresponding amino acids of said phosphodiesterase to provide a rough structure of said phosphodiesterase; and constructing structures representing the remainder of said phosphodiesterase using electronic representations of the structures of the remaining amino acid residues in said phosphodiesterase.

49. The method of claim 48, further comprising fitting said homology model to low resolution x-ray diffraction data from one or more crystals of said phosphodiesterase.

50. The method of claim 48, wherein the coordinates of conserved residues from Table 1 or 2 are utilized.

51. The method of claim 48, wherein said phosphodiesterase is PDE5A.

52. An electronic representation of a crystal structure of PDE4B, containing atomic coordinate representations corresponding to the coordinates listed in Table 1 or 2,

53. The electronic representation of claim 52, comprising a schematic representation.

54. The electronic representation of claim 52, wherein said PDE4B consists essentially of a PDE4B phosphodiesterase domain.

55. An electronic representation of a PDE4B-based homology model for a phosphodiesterase, wherein said homology model utilizes conserved residue atomic coordinates of Table 1 or 2.

56. A method for developing a biological agent, comprising:
analyzing a PDE4B crystal structure and identifying at least one sub-structure for forming said biological agent.

57. The method of claim 56, wherein said substructure comprises an epitope, and said method further comprises developing antibodies against said epitope.

58. The method of claim 56, wherein said sub-structure comprises a mutation site expected to provide altered activity, and said method further comprises creating a mutation at said site thereby providing a modified PDE4B.

59. The method of claim 56, wherein said sub-structure comprises an attachment point for attaching a separate moiety.

60. The method of claim 56, wherein said separate moiety is selected from the group consisting of a peptide, a polypeptide, a solid phase material, a linker, and a label.

61. The method of claim 56, further comprising attaching said separate moiety.

62. A method for identifying compounds with the potential to bind PDE4B, comprising:
fitting at least one electronic representation of a compound in an electronic representation of a PDE4B binding site, wherein said binding site includes electronic representations of coordinates of conserved binding site residues from Table 4.

63. The method of claim 62, wherein said electronic representation further comprises an electronic representation of a binding compound complexed in said binding site.

64. The method of claim 63, comprising:
removing a computer representation of a compound complexed with PDE4B and fitting a computer representation of a compound from a computer database with a computer representation of the active site of PDE4B; and identifying compounds that best fit said active site based on favorable geometric fit and energetically favorable complementary interactions as potential binding compounds.

65. The method of claim 63, comprising:
modifying a computer representation of a compound complexed with PDE4B by the deletion or addition or both of one or more chemical groups;
fitting a computer representation of a compound from a computer database with a computer representation of the active site of PDE4B; and identifying compounds that best fit said active site based on favorable geometric fit and energetically favorable complementary interactions as potential binding compounds.

66. The method of claim 63, comprising:
removing a computer representation of a compound complexed with PDE4B; and searching a database for compounds having structural similarity to said compound using a compound searching computer program or replacing portions of said compound with similar chemical structures using a compound construction computer program.

67. The method of claim 63, wherein said compound complexed with PDE4B is non-hydrolyzable cGMP analog.

68. The method of claim 62, wherein said fitting comprises determining whether said compounds will interact with one or more of conserved PDE4B active site residues.

69. A method for attaching a PDE4B binding compound or derivative thereof to an attachment component so as to maintain substantial PDE4B binding of said PDE4B
binding compound, comprising:
attaching said PDE4B binding compound or derivative thereof to said attachment component at an energetically allowed site for attachment.

70. The method of claim 69, wherein said attachment component is a linker for attachment to a solid phase medium, and said method further comprises attaching said compound or derivative to a solid phase medium through a linker attached at said energetically allowed site.

71. The method of claim 69, wherein said phosphodiesterase comprises conserved residues matching at least one conserved PDE4B active site residues.

72. The method of claim 70, wherein said linker is a traceless linker.

73. The method of claim 70, wherein said phosphodiesterase binding compound or derivative thereof is synthesized on said linker attached to said solid phase medium.

74. The method of claim 73, wherein a plurality of said compounds or derivatives are synthesized in combinatorial synthesis.

75. The method of claim 70, wherein attachment of said compound to said solid phase medium provides an affinity medium.

76. The method of claim 69, wherein said attachment component comprises a label.

77. The method of claim 76, wherein said label comprises a fluorophore.

78. A modified compound, comprising a PDE4B binding compound, with a linker moiety attached thereto at an energetically allowed site for binding of said modified compound to PDE4B.

79. The compound of claim 78, wherein said linker is attached to a solid phase.

80. The compound of claim 78, wherein said linker comprises or is attached to a label.

81. The compound of claim 78, wherein said linker is a traceless linker.

82. The compound of claim 78, wherein said binding compound is a compound for Formula I.

83. A method for developing a ligand for a phosphodiesterase, said phosphodiesterase comprising conserved residues matching one or more conserved active site residues, comprising:
determining whether a PDE4B binding compound which binds to said phosphodiesterase interacts with said one or more conserved PDE4B active site residues in a crystal structure.

84. The method of claim 83, wherein said phosphodiesterase comprises conserved residues matching at least two conserved PDE4B active site residues.

85. The method of claim 83, further comprising determining whether said compound modulates said phosphodiesterase.

86. The method of claim 83, wherein said determining comprises computer fitting said compound in a binding site of said phosphodiesterase.

87. The method of claim 83, further comprising forming a co-crystal of said phosphodiesterase and said compound.

88. The method of claim 83, further comprising determining the binding orientation of said compound with said phosphodiesterase.

89. The method of claim 83, wherein said binding compound is a compound of Formula I.

90. A method for identifying a compound having selectivity between PDE4B and PDE4D, comprising:

analyzing whether a compound differentially interacts in PDE4B and PDE4D in at least one differential site, wherein a differential interaction is indicative of said selectivity.

91. The method of claim 90, Wherein said analyzing comprises:
fitting an electronic representation of said compound in electronic representations of binding sites of PDE4B and PDE4D; and determining whether said compound differentially interacts based on said fitting.

92. The method of claim 90, comprising:
fitting an electronic representation of an initial compound to electronic representations of binding sites of PDE4B and PDE4D, wherein said initial compound binds both and PDE4D;
modifying said electronic representation of said initial compound with at least one moiety that interacts with at least differential site; and determining whether the modified compound differentially binds to PDE4B and PDE4D,

93. The method of claim 92, wherein said modified compound binds differentially to a greater extent than does said initial compound.

94. The method of claim 90, further comprising assaying a compound that differentially interacts for differential activity on PDE4B and PDE4D.

95. The method of claim 90, wherein said compound is a compound of Formula I.

96. A compound having the chemical structure all salts, prodrugs, tautomers and isomers thereof, wherein:
k is selected from the group consisting of -CR6R7R19, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14;

Z is O, S, or NR9;
t is N or CH;
y is N or CH, provided that both t and y are not N;
v, w and x are CH;
A has a structure selected from the group consisting of R6 and R7 are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylakyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;

R8 at each occurrence is independently selected from the group consisting of -OR9, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R8 is alkenyl, no alkene carbon thereof is bound to C(Z), optionally substituted lower alkynyl, provided, however, that when R8 is alkynyl, no alkyne carbon thereof is bound to C(Z), optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R9 at each occurrence is independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R9 is alkenyl, no alkene carbon thereof is bound to O, N or S, optionally substituted lower alkynyl, provided, however, that when R9 is alkynyl, no alkyne carbon thereof is bound to O, N or S, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R10 and R11 at each occurence are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R10 and/or R11 are alkenyl, no alkene carbon thereof is bound to nitrogen, optionally substituted lower alkynyl, provided, however, that when R10 and/or R11 are alkynyl, no alkyne carbon thereof is bound to nitrogen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14; or R10 and R11 together with the nitrogen to which they are attached form a 5-7 membered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring;
R12 and R13 at each occurrence are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R12 and/or R13 are alkenyl, no alkene carbon thereof is bound to nitrogen, optionally substituted lower alkynyl, provided, however, that when R12 and/or R13 are alkynyl, no alkyne carbon thereof is bound to nitrogen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl; or R12 and R13 together with the nitrogen to which they are attached form a 5-7 membered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring;
R14 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R14 is alkenyl, no alkene carbon thereof is bound to -S(O)2-, optionally substituted lower alkynyl, provided, however, that when R14 is alkynyl, no alkyne carbon thereof is bound to -S(O)2-, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R15 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R15 is alkenyl, no alkene carbon thereof is bound to oxygen, optionally substituted lower alkynyl, provided, however, that when R15 is alkynyl, no alkyne carbon thereof is bound to oxygen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, and -C(Z)NR12R13; or each R15, along with the oxygens to which they are bound, combine to form a 5-membered optionally substituted heterocycloalkyl ring fused to the phenyl ring;
R16 is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl provided, however, that when R-16 is alkenyl, no alkene carbon thereof is bound to oxygen, optionally substituted lower alkynyl provided, however, that when R16 is alkynyl, no alkyne carbon thereof is bound to oxygen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, and -C(Z)NR12R13;

R17 is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl provided, however, that when R17 is alkenyl, no alkene carbon thereof is bound to N, O, or S, optionally substituted lower alkynyl, provided, however, that when R17 is alkynyl, no alkyne carbon thereof is bound to N, O, or S, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R18 is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, -OR9, -SR9, -NR10R11, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14; and R19 is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
provided, however that the compound is not

97. A compound having the chemical structure selected from the group consisting of all salts, prodrugs, tautomers and isomers thereof, wherein:
k is selected from the group consisting of -CR6R7R19, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14;
Z is O, S, or NR9;
t is N or CH;
y is N or CH, provided that both t and y are not N;
u, v, w and x are CH;
A has a structure selected from the group consisting of R6 and R7 are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylakyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R8 at each occurrence is independently selected from the group consisting of -OR9, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R8 is alkenyl, no alkene carbon thereof is bound to C(Z), optionally substituted lower alkynyl, provided, however, that when R8 is alkynyl, no alkyne carbon thereof is bound to C(Z), optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R9 at each occurrence is independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R9 is alkenyl, no alkene carbon thereof is bound to O, N or S, optionally substituted lower alkynyl, provided, however, that when R9 is alkynyl, no alkyne carbon thereof is bound to O, N or S, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R10 and R11 at each occurrence are independently selected from, the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R10 and/or R11 are alkenyl, no alkene carbon thereof is bound to nitrogen, optionally substituted lower alkynyl, provided, however, that when R10 and/or R11 are alkynyl, no alkyne carbon thereof is bound to nitrogen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, -C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14; or R10 and R11 together with the nitrogen to which they are attached form a 5-7 membered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring;
R12 and R13 at each occurrence are independently selected from the group consisting of hydrogen, optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R12 and/or R13 are alkenyl, no alkene carbon thereof is bound to nitrogen, optionally substituted lower alkynyl, provided, however, that when R12 and/or R13 are alkynyl, no alkyne carbon thereof is bound to nitrogen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl; or R12 and R13 together with the nitrogen to which they are attached form a 5-7 membered optionally substituted heterocycloalkyl or optionally substituted heteroaryl ring;
R14 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R14 is alkenyl, no alkene carbon thereof is bound to -S(O)2-, optionally substituted lower alkynyl, provided, however, that when R14 is alkynyl, no alkyne carbon thereof is bound to -S(O)2-, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl;
R15 at each occurrence is independently selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, provided, however, that when R15 is alkenyl, no alkene carbon thereof is bound to oxygen, optionally substituted lower alkynyl, provided, however, that when R15 is alkynyl, no alkyne carbon thereof is bound to oxygen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, and -C(Z)NR12R13; or each R15, along with the oxygens to which they are bound, combine to form a 5-membered optionally substituted heterocycloalkyl ring fused to the phenyl ring;
R16 is selected from the goup consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl provided, however, that when R16 is alkenyl, no alkene carbon thereof is bound to oxygen, optionally substituted lower alkynyl provided, however, that when R16 is alkynyl, no alkyne carbon thereof is bound to oxygen, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, and -C(Z)NR12R13;
R17 is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl provided, however, that when R17 is alkenyl, no alkene carbon thereof is bound to N, O, or S, optionally substituted lower alkynyl, provided, however, that when R17 is alkynyl, no alkyne carbon thereof is bound to N, O, or S, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl; and optionally substituted heteroaralkyl;
R18 is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -C(Z)R8, -OR9, -SR9, -NR10R11, C(Z)NR12R13, -S(O)2NR12R13, and -S(O)2R14; and R19is selected from the group consisting of optionally substituted lower alkyl, optionally substituted lower alkenyl, optionally substituted lower alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, and optionally substituted heteroaralkyl.

98. ~The compound of claim 97, wherein t and y are CH.

99. ~The compound of claim 97, wherein t is CH and y is N.