CA2628883A1 - Novel 3'-modified oligonucleotide derivatives - Google Patents
Novel 3'-modified oligonucleotide derivatives Download PDFInfo
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- CA2628883A1 CA2628883A1 CA 2628883 CA2628883A CA2628883A1 CA 2628883 A1 CA2628883 A1 CA 2628883A1 CA 2628883 CA2628883 CA 2628883 CA 2628883 A CA2628883 A CA 2628883A CA 2628883 A1 CA2628883 A1 CA 2628883A1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
- C07C69/34—Esters of acyclic saturated polycarboxylic acids having an esterified carboxyl group bound to an acyclic carbon atom
- C07C69/40—Succinic acid esters
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
Abstract
3'-Derivatised oligonucleotide analogues of formula (I) and (II) and their salts are new: R<1>-(A<1>)n'-(B<1>)m'-(A<2>)n-(B<2>)m-B<3> (I); R<1>-(A<1>)n'-(B<1>)m'-(A<2>)n-A<3> (II); A<1> and A<2> = gps. of formula (i); B<1> and B<2> = gps. of formula (ii); B<3> = a gp. of formula (iii); and A<3> = a gp. of formula (iv); a and b = 0 to 20; R<1> = H, 1-18C alkyl, 2-18C alkenyl, 3-18C alkynyl, 2-19C alkanoyl, 3-20C alkenoyl, 4-20C alkynoyl, 6-20C aryl, (6-14C)aryl(1-8C)alkyl or P(W)ZZ'; R<2> = H, OH, 1-18C alkoxy, halogen, azido or NH2; D = OH or OPO3<2->; B = a natural or unnatural nucleotide base; n = 1-100; n' = 0-50; m = 0-5; m' = 0-5 in (I) or 1-5 in (II); A = O, S or CH2; W = O, S or Se; V = O or S; T = O, S or NH; Y = O, S, NH or CH2; X = OH or SH; U = OH, SH, BH3 (sic), SeH, 1-18C alkoxy, 1-18C alkyl, 6-20C aryl, (6-14C)aryl(1-8C)alkyl, NHR<3>, NR<3>R<4> or (OCH2CH2)pO(CH2)qCH2R<5>; R<3> = 1-18C alkyl, 6-20C aryl, (6-14C)aryl(1-8C)alkyl or (CH2)c(NH(CH2)c)dNR<6>R<6>; c = 2-6; d = 0-6; R<6> = H, 1-6C alkyl or (1-4C)alkoxy(1-6C)alkyl; R<4> = 1-18C alkyl, 6-20C aryl or (6-14C)aryl(1-8C)alkyl; or NR<3>R<4> = a 5- or 6-membered ring opt. contg. another N, O or S atom; p = 1-100; q = 0-22; R<5> = H or a functional gp. such as OH, NH2, NHR<7>, COOH, CONH2, COOR<8> or halogen; R<7> = 1-6C alkyl, R<8> = 1-4C alkyl; Z and Z' = OH, SH, SeH, 1-22C alkoxy, O(CH2)bNHR<7>'R<8>', 1-18C alkyl, 6-20C aryl, (6-14)aryl(1-8C)alkyl (where aryl includes heteroaryl and is opt. substd. by 1-3 of COOH, NH2, NO2, 1-4C alkylamino, 1-6C alkoxy, OH, halogen and CN), 1-18C alkylthio, NHR<3>, NR<3>R<4>, a 3'- or 5'-linked nucleoside or oligonucleotide, or a gp. which enhances intracellular uptake, serves as a DNA probe label or binds to, crosslinks with or cleaves a target nucleic acid after hybridisation; R<7>'R<8>' = as R<7>, R<8>; or NR<7>'R<8> = a 3- to 6-membered ring.
Claims (16)
1. A compound of the formula II:
and the physiologically tolerated salts thereof, in which a is an integer from zero to 20;
b is an integer from zero to 20;
R1 is hydrogen, C1-C18-alkyl, C2-C18-alkenyl, C3-C18-alkynyl, C1-C18-alkylcarbonyl, C2-C19-alkenylcarbonyl, C3-C19-alkynylcarbonyl, C6-C20-aryl, C6-C14-aryl-C1-C8-alkyl, or a radical of the formula III
R2 is hydrogen, hydroxyl, C1-C18-alkoxy, halogen, azido or NH2;
D is hydroxyl or O-PO3 2;
B is a natural or unnatural base customary in nucleotide chemistry;
n is an integer from 7 to 25;
n' is an integer from zero to 50;
m' in is an integer from 1 to 5;
A is oxy, thioxy or methylene;
W is oxo, thioxo or selenoxo;
V is oxo or thio;
T is oxy, thio or imino;
Y is oxy, thio, imino or methylene;
X is hydroxyl or mercapto;
U is hydroxyl, mercapto, BH3, SeH, C1-C18-alkoxy, C1-C18-alkyl, C6-C20-aryl, C6-C14-aryl-C1-C8-alkyl, NHR3, NR3R4 or a radical of the formula IV
(OCH2CH2)p O(CH2)q CH2R5 (IV) in which R3 is C1-C18-alkyl, C6-C20-aryl, C6-C14-aryl-C1-C8-alkyl or -(CH2)c-[NH(CH2)c]d-NR6R6, in which c is an integer from 2 to 6 and d is an integer from zero to 6 , and R6 is independently of one another, hydrogen, C1-C6-alkyl or C1-C4-alkoxy-C1-C6-alkyl;
R4 is C1-C18-alkyl, C6-C20-aryl or C6-C10-aryl-C1-C8-alkyl, or, in the case of NR3R9, together with R3 and the nitrogen atom carrying them is a 5-6-membered heterocyclic ring which can additionally contain another heteroatom selected from the series consisting of O, S
and N;
p is an integer from 1 to 100;
q is an integer from zero to 22;
R5 is hydrogen, hydroxyl, amino, NHR7, COOH, CONH2. COOR8 or halogen, in which R7 is C1-C6-alkyl and R8 is C1-C4-alkyl;
Z, Z' are, independently of one another, hydroxyl;
mercapto; SeH; C1-C22-alkoxy; -O-(CH2)b'-NR7R8, in which b' is an integer from 1 to 6, and R7 is C1-C6-alkyl and R8 is C1-C4-alkyl, or R7 and R8 form, together with the nitrogen atom carrying them, a 3-6-membered ring; C1-C18-alkyl; C6-C20-aryl, C6-C14-aryl-C1-C8-alkyl, C6-C14-aryl-C1-C8-alkoxy, C6-C20-heteroaryl, C6-C14-heteroaryl-C1-C8-alkyl, or C6-C14-heteroaryl-C1-C8-alkoxy in which aryl and heteroaryl are unsubstituted or substituted by 1, 2 or 3 identical or different radicals selected from the series consisting of carboxyl, amino, nitro, C1-C4-alkylamino, C1-C6-alkoxy, hydroxyl, halogen and cyano; C1-C18-alkylmercapto; NHR3 or NR3R4 in which R3 and R4 are as defined above; or a group a) which favours intracellular uptake selected from -O-(CH2)x-CH3, in which x is an integer from 8 to 18;
-O-(CH2)e-CH=CH-(CH2)f-CH3, in which e and f independently of one another are an integer from 6 to 12;
-O-(CH2CH2O)4-(CH2)9-CH3; -O-(CH2CH2O)8-(CH2)13-CH3; and -O-(CH2CH2O)7-(CH2)15-CH3; steroid residues and conjugates which utilize natural carrier systems; conjugates of mannose; and conjugates of peptides of the appropriate receptors which lead to receptor mediated endocytosis of the compounds of formula II; or b)which acts as a labelling group selected from fluorescent groups; chemiluminescent groups; and linker groups having functional groups which permit subsequent derivatization with detectable reporter groups; or c)on hybridization of the compound of formula II onto a target nucleic acid, interacts with the target nucleic acid by binding, crosslinking or cleavage; or d) which is a nucleoside or oligonucleotide linked via the 5' or 3' end;
and the curved parenthesis indicates that R2 and the adjacent phosphoryl radical can be located in the 2' and 3' positions or else conversely in the 3' and 2' positions, each nucleotide is in its D or L configuration and the base B is located in the .alpha. or .beta. position.
and the physiologically tolerated salts thereof, in which a is an integer from zero to 20;
b is an integer from zero to 20;
R1 is hydrogen, C1-C18-alkyl, C2-C18-alkenyl, C3-C18-alkynyl, C1-C18-alkylcarbonyl, C2-C19-alkenylcarbonyl, C3-C19-alkynylcarbonyl, C6-C20-aryl, C6-C14-aryl-C1-C8-alkyl, or a radical of the formula III
R2 is hydrogen, hydroxyl, C1-C18-alkoxy, halogen, azido or NH2;
D is hydroxyl or O-PO3 2;
B is a natural or unnatural base customary in nucleotide chemistry;
n is an integer from 7 to 25;
n' is an integer from zero to 50;
m' in is an integer from 1 to 5;
A is oxy, thioxy or methylene;
W is oxo, thioxo or selenoxo;
V is oxo or thio;
T is oxy, thio or imino;
Y is oxy, thio, imino or methylene;
X is hydroxyl or mercapto;
U is hydroxyl, mercapto, BH3, SeH, C1-C18-alkoxy, C1-C18-alkyl, C6-C20-aryl, C6-C14-aryl-C1-C8-alkyl, NHR3, NR3R4 or a radical of the formula IV
(OCH2CH2)p O(CH2)q CH2R5 (IV) in which R3 is C1-C18-alkyl, C6-C20-aryl, C6-C14-aryl-C1-C8-alkyl or -(CH2)c-[NH(CH2)c]d-NR6R6, in which c is an integer from 2 to 6 and d is an integer from zero to 6 , and R6 is independently of one another, hydrogen, C1-C6-alkyl or C1-C4-alkoxy-C1-C6-alkyl;
R4 is C1-C18-alkyl, C6-C20-aryl or C6-C10-aryl-C1-C8-alkyl, or, in the case of NR3R9, together with R3 and the nitrogen atom carrying them is a 5-6-membered heterocyclic ring which can additionally contain another heteroatom selected from the series consisting of O, S
and N;
p is an integer from 1 to 100;
q is an integer from zero to 22;
R5 is hydrogen, hydroxyl, amino, NHR7, COOH, CONH2. COOR8 or halogen, in which R7 is C1-C6-alkyl and R8 is C1-C4-alkyl;
Z, Z' are, independently of one another, hydroxyl;
mercapto; SeH; C1-C22-alkoxy; -O-(CH2)b'-NR7R8, in which b' is an integer from 1 to 6, and R7 is C1-C6-alkyl and R8 is C1-C4-alkyl, or R7 and R8 form, together with the nitrogen atom carrying them, a 3-6-membered ring; C1-C18-alkyl; C6-C20-aryl, C6-C14-aryl-C1-C8-alkyl, C6-C14-aryl-C1-C8-alkoxy, C6-C20-heteroaryl, C6-C14-heteroaryl-C1-C8-alkyl, or C6-C14-heteroaryl-C1-C8-alkoxy in which aryl and heteroaryl are unsubstituted or substituted by 1, 2 or 3 identical or different radicals selected from the series consisting of carboxyl, amino, nitro, C1-C4-alkylamino, C1-C6-alkoxy, hydroxyl, halogen and cyano; C1-C18-alkylmercapto; NHR3 or NR3R4 in which R3 and R4 are as defined above; or a group a) which favours intracellular uptake selected from -O-(CH2)x-CH3, in which x is an integer from 8 to 18;
-O-(CH2)e-CH=CH-(CH2)f-CH3, in which e and f independently of one another are an integer from 6 to 12;
-O-(CH2CH2O)4-(CH2)9-CH3; -O-(CH2CH2O)8-(CH2)13-CH3; and -O-(CH2CH2O)7-(CH2)15-CH3; steroid residues and conjugates which utilize natural carrier systems; conjugates of mannose; and conjugates of peptides of the appropriate receptors which lead to receptor mediated endocytosis of the compounds of formula II; or b)which acts as a labelling group selected from fluorescent groups; chemiluminescent groups; and linker groups having functional groups which permit subsequent derivatization with detectable reporter groups; or c)on hybridization of the compound of formula II onto a target nucleic acid, interacts with the target nucleic acid by binding, crosslinking or cleavage; or d) which is a nucleoside or oligonucleotide linked via the 5' or 3' end;
and the curved parenthesis indicates that R2 and the adjacent phosphoryl radical can be located in the 2' and 3' positions or else conversely in the 3' and 2' positions, each nucleotide is in its D or L configuration and the base B is located in the .alpha. or .beta. position.
2. A compound of the formula II as claimed in claim 1, and the physiologically tolerated salts thereof, wherein the base B is located in the .beta. position, the nucleotides are in the D configuration, and R2 is located in position 2'.
3. A compound of the formula II as claimed in claim 1 or 2, and the physiologically tolerated salts thereof, wherein a is an integer from zero to 10;
b is an integer from zero to 10;
R1 is hydrogen or a radical of the formula III
R2 is hydrogen or hydroxyl;
n' is an integer from zero to 30;
A is oxy;
W is oxo or thioxo;
U is hydroxyl, mercapto, C1-C6-alkoxy, C1-C6-alkyl, NR3R4 or NHR3, in which R3 is C1-C18-alkyl; and R4 is C1-C8-alkyl, C6-C20-aryl or C6-C10-C1-C8-alkyl, or, in the case of NR3R4, together with R3 and the nitrogen atom carrying them is a 5-6-membered heterocyclic ring which can additionally contain another heteroatom selected from the series consisting of O, S
and N.
b is an integer from zero to 10;
R1 is hydrogen or a radical of the formula III
R2 is hydrogen or hydroxyl;
n' is an integer from zero to 30;
A is oxy;
W is oxo or thioxo;
U is hydroxyl, mercapto, C1-C6-alkoxy, C1-C6-alkyl, NR3R4 or NHR3, in which R3 is C1-C18-alkyl; and R4 is C1-C8-alkyl, C6-C20-aryl or C6-C10-C1-C8-alkyl, or, in the case of NR3R4, together with R3 and the nitrogen atom carrying them is a 5-6-membered heterocyclic ring which can additionally contain another heteroatom selected from the series consisting of O, S
and N.
4. A compound of the formula II as claimed in any one of claims 1 to 3, and the physiologically tolerated salts thereof, wherein a is an integer from zero to 4;
b is zero;
R1 is hydrogen;
R2 is hydrogen;
D is hydroxyl;
B is adenine, cytosine, guanine, uracil, thymine,
b is zero;
R1 is hydrogen;
R2 is hydrogen;
D is hydroxyl;
B is adenine, cytosine, guanine, uracil, thymine,
5-propinuracil or 5-propincytosine;
n' is an integer from zero to 25;
m' is 1;
T is oxy;
Y is oxy; and U is hydroxyl or C1-C6-alkyl.
5. A compound of the formula II as claimed in any one of claims 1 to 4, and the physiologically tolerated salts thereof, wherein Z, Z' are, independently of one another, hydroxyl;
mercapto; SeH; C1-C20alkoxy; -O-(CH2)b'-NR7R8, in which b' is an integer from 1 to 6, and R7 is C1-C6-alkyl and R8 is C1-C4-alkyl, or R7 and R8 form, together with the nitrogen atom carrying them, a 3-6-membered ring; C1-C8-alkyl; C6-C20-aryl, C6-C10-aryl-C1-C4-alkyl, C6-C10-aryl-C1-C4-alkoxy, C6-C20-heteroaryl, C6-C10-heteroaryl-C1-C4-alkyl, or C6-C10-heteroaryl-C1-C4-alkoxy in which aryl and heteroaryl are unsubstituted or substituted by 1, 2 or 3 identical or different radicals selected from the series consisting of carboxyl, amino, nitro, C1-C4-alkylamino, C1-C6-alkoxy, hydroxyl, halogen and cyano; C1-C18-alkylmercapto; NHR3 or NR3R4 in which R3 and R4 are as defined in claim 1; or a group a) which favours intracellular uptake selected from -O- (CH2)x-CH3, in which x is an integer from 8 to 18;
-O-(CH2)e-CH=CH-(CH2)f-CH3, in which e and f independently of one another are an integer from 6 to 12;
-O-( CH2CH2O)4-(CH2)9-CH3; -O-(CH2CH2O)8-(CH2)13-CH3; and -O-(CH2CH2O)7-(CH2)15-CH3; steroid residues and conjugates which utilize natural carrier systems; conjugates of mannose; and conjugates of peptides of the appropriate receptors which lead to receptor mediated endocytosis of the compounds of formula II; or b)which acts as a labelling group selected from fluorescent groups; chemiluminescent groups; and linker groups having functional groups which permit subsequent derivatization with detectable reporter groups; or c)on hybridization of the compound of formula II onto a target nucleic acid, interacts with the target nucleic acid by binding, crosslinking or cleavage; or d) which is a nucleoside or oligonucleotide linked via the 5' or 3' end.
n' is an integer from zero to 25;
m' is 1;
T is oxy;
Y is oxy; and U is hydroxyl or C1-C6-alkyl.
5. A compound of the formula II as claimed in any one of claims 1 to 4, and the physiologically tolerated salts thereof, wherein Z, Z' are, independently of one another, hydroxyl;
mercapto; SeH; C1-C20alkoxy; -O-(CH2)b'-NR7R8, in which b' is an integer from 1 to 6, and R7 is C1-C6-alkyl and R8 is C1-C4-alkyl, or R7 and R8 form, together with the nitrogen atom carrying them, a 3-6-membered ring; C1-C8-alkyl; C6-C20-aryl, C6-C10-aryl-C1-C4-alkyl, C6-C10-aryl-C1-C4-alkoxy, C6-C20-heteroaryl, C6-C10-heteroaryl-C1-C4-alkyl, or C6-C10-heteroaryl-C1-C4-alkoxy in which aryl and heteroaryl are unsubstituted or substituted by 1, 2 or 3 identical or different radicals selected from the series consisting of carboxyl, amino, nitro, C1-C4-alkylamino, C1-C6-alkoxy, hydroxyl, halogen and cyano; C1-C18-alkylmercapto; NHR3 or NR3R4 in which R3 and R4 are as defined in claim 1; or a group a) which favours intracellular uptake selected from -O- (CH2)x-CH3, in which x is an integer from 8 to 18;
-O-(CH2)e-CH=CH-(CH2)f-CH3, in which e and f independently of one another are an integer from 6 to 12;
-O-( CH2CH2O)4-(CH2)9-CH3; -O-(CH2CH2O)8-(CH2)13-CH3; and -O-(CH2CH2O)7-(CH2)15-CH3; steroid residues and conjugates which utilize natural carrier systems; conjugates of mannose; and conjugates of peptides of the appropriate receptors which lead to receptor mediated endocytosis of the compounds of formula II; or b)which acts as a labelling group selected from fluorescent groups; chemiluminescent groups; and linker groups having functional groups which permit subsequent derivatization with detectable reporter groups; or c)on hybridization of the compound of formula II onto a target nucleic acid, interacts with the target nucleic acid by binding, crosslinking or cleavage; or d) which is a nucleoside or oligonucleotide linked via the 5' or 3' end.
6. A compound of the formula II as claimed in any one of claims 1 to 5, and the physiologically tolerated salts thereof, wherein the steroid residue is cholesterol.
7. A compound of the formula II as claimed in any one of claims 1 to 5, and the physiologically tolerated salts thereof, wherein the conjugates which utilize natural carrier systems are selected from bile acid, folic acid and 2-(N-alkyl-N-alkoxy)aminoanthraquinone.
8. A compound of the formula II as claimed in any one of claims 1 to 5, and the physiologically tolerated salts thereof, wherein the peptides of the appropriate receptors which lead to receptor-mediated endocyctosis of the compounds of formula II are selected from epidermal growth factor, bradykinin and platelet derived growth factor.
9. A compound of the formula II as claimed in any one of claims 1 to 5, and the physiologically tolerated salts thereof, wherein the fluorescent groups are selected from N-dimethyl-1-aminonaphthyl-5-sulfonyl (dansyl), fluorescein and coumarin derviatives.
10. A compound of the formula II as claimed in any one of claims 1 to 5, and the physiologically tolerated salts thereof, wherein the chemiluminescent groups are selected from acridine derivatives, the digitoxin system and the biotin/avidin system.
11. A process for the preparation of compounds of the formula II as claimed in any one of claims 1-10, which comprises a) eliminating from a compound of the formula XVI
in which B' is B, R2' is R2, S2 is a suitable protective group, SS
is a solid support, Li is a linker arm, and A, B, R2, Y and V are as defined in claim 1 and Li is or is not a linker which permits introduction of a 3' phosphate residue, the protective group S2;
b) subsequently reacting the resulting compound by the phosphoramidate method with a nucleoside phosphoramidite of the formula XI
in which A, B', R2', S2, Y and V are as defined in step a), and B
is in unprotected or protected form, R9 and R10 are identical or different and are C1-C8-alkyl or C5-C12-cycloalkyl, benzyl or phenyl or together with the nitrogen atom to which they are bonded form a saturated or unsaturated heterocyclic ring, with or without further heteroatoms selected from N, O and S and substituents, R12 is OR13 or C1-C18-alkyl, C1-C18-alkoxy, C6-C20-aryl or C6-C14-aryl-C1-C8-alkyl, R13 is a group of the formula or a benzyl group, which is unsubstituted or is one to four times ring-substituted, where the substituent or substituents are, independently of one another, fluorine, chlorine, bromine, a C1-C4-alkyl, nitro, methoxy or carboxyl group, oxidizing the resulting compound, carrying out a capping, eliminating the protective group S2 and then repeating this reaction step (n - 1) times, resulting in a compound of the formula XVII
in which A, B' , Li, R2', SS, Y and V are as defined in step a) and U, W and n are as defined in claim 1;
c) subsequently reacting the resulting compound with a compound of the formula IX
in which S1 is a suitable protective group, V' is V, R9, R10, R12, V and S2 are as defined in step b), and T, a and b are as defined in claim 1, in the presence of a compound of the formula [HNR14R15R16](+)E(-), where R14, R15 and R16 are identical to or different from one another and are a C1-C4-alkyl group and E is fluorine, chlorine, bromine, or in the presence of tetrazole or substituted tetrazole in a suitable organic solvent, oxidizing the resulting compound, carrying out a capping, eliminating the protective group S2, and then repeating this reaction step (m' - 1) times, resulting in a compound of the formula XVIII
in which A, B', Li, R2', SS, U, V, V', Y and n are as defined in step b), Si is a suitable protective group, m' is m and m, a and b are defined in claim 1;
d) if n' is 1-50, carrying out reaction step b), which is repeated (n' - 1) times, resulting in a compound of the formula XIX
in which A, B', Li, R2', S1, SS, U, V, V', W, Y, a, b, m' and n are as defined in step c) and n' is n;
e) if R1 .noteq. H in formula II, introducing the radical R1 into the compound obtained in c) or d), where R1 is C1-C18-alkyl, C2-C18-alkenyl, C3-C18-alkynyl, C1-C18-alkylcarbonyl, C2-C19-alkenylcarbonyl, C3-C19-, C6-C20-aryl, C6-C14-aryl-C1-C8-alkyl, or a radical of the formula III
in which W is as defined in step b) and Z and Z' are as defined in claim 1;
f) if R1 = H in formula II, capping the compound obtained in c) or d);
g) subsequently eliminating the protective group S1 so that the linker to the solid support and the other protective groups present in the molecule are retained;
h) and reacting the compound obtained in this way with a compound of the formula XV
in which R9, R10 and R12 are as defined in step b) , and Z" has the meaning of Z as defined in step e) or else Z is protected, in the presence of a compound of the formula [HNR14R15R16](+)E(-), where R14, R15, R16 and E are as defined in step c), or in the presence of tetrazole or substituted tetrazole in a suitable organic solvent;
i) oxidizing the compound obtained in step h);
j) eliminating the compound obtained in step i) from the support, and eliminating the remaining protective groups on the phosphate and nucleotide bases.
in which B' is B, R2' is R2, S2 is a suitable protective group, SS
is a solid support, Li is a linker arm, and A, B, R2, Y and V are as defined in claim 1 and Li is or is not a linker which permits introduction of a 3' phosphate residue, the protective group S2;
b) subsequently reacting the resulting compound by the phosphoramidate method with a nucleoside phosphoramidite of the formula XI
in which A, B', R2', S2, Y and V are as defined in step a), and B
is in unprotected or protected form, R9 and R10 are identical or different and are C1-C8-alkyl or C5-C12-cycloalkyl, benzyl or phenyl or together with the nitrogen atom to which they are bonded form a saturated or unsaturated heterocyclic ring, with or without further heteroatoms selected from N, O and S and substituents, R12 is OR13 or C1-C18-alkyl, C1-C18-alkoxy, C6-C20-aryl or C6-C14-aryl-C1-C8-alkyl, R13 is a group of the formula or a benzyl group, which is unsubstituted or is one to four times ring-substituted, where the substituent or substituents are, independently of one another, fluorine, chlorine, bromine, a C1-C4-alkyl, nitro, methoxy or carboxyl group, oxidizing the resulting compound, carrying out a capping, eliminating the protective group S2 and then repeating this reaction step (n - 1) times, resulting in a compound of the formula XVII
in which A, B' , Li, R2', SS, Y and V are as defined in step a) and U, W and n are as defined in claim 1;
c) subsequently reacting the resulting compound with a compound of the formula IX
in which S1 is a suitable protective group, V' is V, R9, R10, R12, V and S2 are as defined in step b), and T, a and b are as defined in claim 1, in the presence of a compound of the formula [HNR14R15R16](+)E(-), where R14, R15 and R16 are identical to or different from one another and are a C1-C4-alkyl group and E is fluorine, chlorine, bromine, or in the presence of tetrazole or substituted tetrazole in a suitable organic solvent, oxidizing the resulting compound, carrying out a capping, eliminating the protective group S2, and then repeating this reaction step (m' - 1) times, resulting in a compound of the formula XVIII
in which A, B', Li, R2', SS, U, V, V', Y and n are as defined in step b), Si is a suitable protective group, m' is m and m, a and b are defined in claim 1;
d) if n' is 1-50, carrying out reaction step b), which is repeated (n' - 1) times, resulting in a compound of the formula XIX
in which A, B', Li, R2', S1, SS, U, V, V', W, Y, a, b, m' and n are as defined in step c) and n' is n;
e) if R1 .noteq. H in formula II, introducing the radical R1 into the compound obtained in c) or d), where R1 is C1-C18-alkyl, C2-C18-alkenyl, C3-C18-alkynyl, C1-C18-alkylcarbonyl, C2-C19-alkenylcarbonyl, C3-C19-, C6-C20-aryl, C6-C14-aryl-C1-C8-alkyl, or a radical of the formula III
in which W is as defined in step b) and Z and Z' are as defined in claim 1;
f) if R1 = H in formula II, capping the compound obtained in c) or d);
g) subsequently eliminating the protective group S1 so that the linker to the solid support and the other protective groups present in the molecule are retained;
h) and reacting the compound obtained in this way with a compound of the formula XV
in which R9, R10 and R12 are as defined in step b) , and Z" has the meaning of Z as defined in step e) or else Z is protected, in the presence of a compound of the formula [HNR14R15R16](+)E(-), where R14, R15, R16 and E are as defined in step c), or in the presence of tetrazole or substituted tetrazole in a suitable organic solvent;
i) oxidizing the compound obtained in step h);
j) eliminating the compound obtained in step i) from the support, and eliminating the remaining protective groups on the phosphate and nucleotide bases.
12. The process according to claim 11, wherein the protective group S2 is dimethoxytrityl or monomethoxytrityl.
13. The process as claimed in claim 11 or 12, wherein the linker arm Li is an aminoalkyl linker.
14. The process as claimed in claim 13 wherein the aminoalkyl linker is derivatized by reaction with an acridinium active ester.
15. A use of the compounds of the formula II as claimed in any one of claims 1 to 10 as a probe for detecting nucleic acids.
16. A pharmaceutical composition comprising a compound of the formula II as claimed in any one of claims 1 to 10 and a physiologically tolerated ancillary substance and/or vehicle.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DEP4420737.9 | 1994-06-15 | ||
| DE4420737A DE4420737A1 (en) | 1994-06-15 | 1994-06-15 | New 3'-derivatised oligo:nucleotide analogues |
| DE4424263A DE4424263A1 (en) | 1994-07-09 | 1994-07-09 | New 3'-derivatised oligo:nucleotide analogues |
| DEP4424263.8 | 1994-07-09 | ||
| CA 2151801 CA2151801C (en) | 1994-06-15 | 1995-06-14 | 3'-modified oligonucleotide derivatives |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2151801 Division CA2151801C (en) | 1994-06-15 | 1995-06-14 | 3'-modified oligonucleotide derivatives |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2628883A1 true CA2628883A1 (en) | 1995-12-16 |
| CA2628883C CA2628883C (en) | 2011-02-15 |
Family
ID=25937419
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2151801 Expired - Fee Related CA2151801C (en) | 1994-06-15 | 1995-06-14 | 3'-modified oligonucleotide derivatives |
| CA 2628883 Expired - Fee Related CA2628883C (en) | 1994-06-15 | 1995-06-14 | Novel 3'-modified oligonucleotide derivatives |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2151801 Expired - Fee Related CA2151801C (en) | 1994-06-15 | 1995-06-14 | 3'-modified oligonucleotide derivatives |
Country Status (9)
| Country | Link |
|---|---|
| US (1) | US5696248A (en) |
| EP (1) | EP0688784B1 (en) |
| JP (1) | JP4398517B2 (en) |
| AT (1) | ATE312840T1 (en) |
| AU (1) | AU701333B2 (en) |
| CA (2) | CA2151801C (en) |
| DE (1) | DE59511027D1 (en) |
| DK (1) | DK0688784T3 (en) |
| ES (1) | ES2253740T3 (en) |
Families Citing this family (24)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6291438B1 (en) | 1993-02-24 | 2001-09-18 | Jui H. Wang | Antiviral anticancer poly-substituted phenyl derivatized oligoribonucleotides and methods for their use |
| US6207646B1 (en) | 1994-07-15 | 2001-03-27 | University Of Iowa Research Foundation | Immunostimulatory nucleic acid molecules |
| US5855911A (en) * | 1995-08-29 | 1999-01-05 | Board Of Regents, The University Of Texas System | Liposomal phosphodiester, phosphorothioate, and P-ethoxy oligonucleotides |
| US7309692B1 (en) * | 1996-07-08 | 2007-12-18 | Board Of Regents, The University Of Texas System | Inhibition of chronic myelogenous leukemic cell growth by liposomal-antisense oligodeoxy-nucleotides targeting to GRB2 or CRK1 |
| US6977244B2 (en) * | 1996-10-04 | 2005-12-20 | Board Of Regents, The University Of Texas Systems | Inhibition of Bcl-2 protein expression by liposomal antisense oligodeoxynucleotides |
| US6723706B2 (en) | 1997-05-05 | 2004-04-20 | Aventis Pharma Deutschland Gmbh | Modified antisense nucleotides complementary to a section of the human Ha-ras gene |
| US7285288B1 (en) | 1997-10-03 | 2007-10-23 | Board Of Regents, The University Of Texas System | Inhibition of Bcl-2 protein expression by liposomal antisense oligodeoxynucleotides |
| US7704962B1 (en) | 1997-10-03 | 2010-04-27 | Board Of Regents, The University Of Texas System | Small oligonucleotides with anti-tumor activity |
| US20040186071A1 (en) | 1998-04-13 | 2004-09-23 | Bennett C. Frank | Antisense modulation of CD40 expression |
| US5968826A (en) * | 1998-10-05 | 1999-10-19 | Isis Pharmaceuticals Inc. | Antisense inhibition of integrin α4 expression |
| WO2000040592A1 (en) * | 1998-12-30 | 2000-07-13 | Oligos Etc. Inc. | Acid stable backbone modified end-blocked nucleic acids and therapeutic uses thereof |
| DE10019252A1 (en) * | 2000-04-18 | 2001-10-31 | Klaus Karl Degitz | Polydeoxyribonucleotides to inhibit ICAM-1 gene expression |
| CA2471967A1 (en) * | 2002-01-03 | 2003-07-31 | Board Of Regents, The University Of Texas System | Wt1 antisense oligos for the inhibition of breast cancer |
| GB0209539D0 (en) * | 2002-04-26 | 2002-06-05 | Avecia Ltd | Monomer Polymer and process |
| AR040996A1 (en) | 2002-08-19 | 2005-04-27 | Coley Pharm Group Inc | IMMUNE STIMULATING NUCLEIC ACIDS |
| AU2003300919A1 (en) | 2002-12-11 | 2004-06-30 | Coley Pharmaceutical Gmbh | 5' cpg nucleic acids and methods of use |
| CA2520538C (en) | 2003-03-31 | 2014-04-29 | F. Hoffmann-La Roche Ag | Compositions and methods for detecting certain flaviviruses, including members of the japanese encephalitis virus serogroup |
| CA2540949A1 (en) | 2003-10-30 | 2005-05-12 | Coley Pharmaceutical Gmbh | C-class oligonucleotide analogs with enhanced immunostimulatory potency |
| JP2008516991A (en) | 2004-10-20 | 2008-05-22 | アンチセンス セラピューティクス リミテッド | Antisense regulation of integrin α4 expression |
| MY159370A (en) | 2004-10-20 | 2016-12-30 | Coley Pharm Group Inc | Semi-soft-class immunostimulatory oligonucleotides |
| MX2009003398A (en) | 2006-09-27 | 2009-08-12 | Coley Pharm Gmbh | Cpg oligonucleotide analogs containing hydrophobic t analogs with enhanced immunostimulatory activity. |
| EP2222851B1 (en) | 2007-11-20 | 2017-06-28 | Ionis Pharmaceuticals, Inc. | Modulation of cd40 expression |
| US8889890B2 (en) | 2009-09-11 | 2014-11-18 | Segetis, Inc. | Ketal esters of oxocarboxylic acids and process of making |
| WO2013120003A1 (en) | 2012-02-08 | 2013-08-15 | Isis Pharmaceuticals, Inc. | Modulation of rna by repeat targeting |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR930016437A (en) * | 1992-01-22 | 1993-08-26 | 귀틀라인, 슈미트 | Oligonucleotide Analogues, Methods for Making and Uses thereof |
| US5646261A (en) * | 1992-01-22 | 1997-07-08 | Hoechst Aktiengesellschaft | 3'-derivatized oligonucleotide analogs with non-nucleotidic groupings, their preparation and use |
-
1995
- 1995-06-05 US US08/462,305 patent/US5696248A/en not_active Expired - Lifetime
- 1995-06-09 AT AT95108896T patent/ATE312840T1/en active
- 1995-06-09 ES ES95108896T patent/ES2253740T3/en not_active Expired - Lifetime
- 1995-06-09 EP EP19950108896 patent/EP0688784B1/en not_active Expired - Lifetime
- 1995-06-09 DE DE59511027T patent/DE59511027D1/en not_active Expired - Lifetime
- 1995-06-09 DK DK95108896T patent/DK0688784T3/en active
- 1995-06-13 AU AU21659/95A patent/AU701333B2/en not_active Ceased
- 1995-06-14 CA CA 2151801 patent/CA2151801C/en not_active Expired - Fee Related
- 1995-06-14 JP JP17043695A patent/JP4398517B2/en not_active Expired - Fee Related
- 1995-06-14 CA CA 2628883 patent/CA2628883C/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| DK0688784T3 (en) | 2006-04-18 |
| AU701333B2 (en) | 1999-01-28 |
| CA2628883C (en) | 2011-02-15 |
| EP0688784A2 (en) | 1995-12-27 |
| AU2165995A (en) | 1995-12-21 |
| US5696248A (en) | 1997-12-09 |
| DE59511027D1 (en) | 2006-01-19 |
| JPH0827178A (en) | 1996-01-30 |
| CA2151801A1 (en) | 1995-12-16 |
| EP0688784B1 (en) | 2005-12-14 |
| ATE312840T1 (en) | 2005-12-15 |
| ES2253740T3 (en) | 2006-06-01 |
| CA2151801C (en) | 2008-08-19 |
| JP4398517B2 (en) | 2010-01-13 |
| EP0688784A3 (en) | 1997-01-22 |
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