US20040115623A1 - Method for treating herpes viruses - Google Patents

Method for treating herpes viruses Download PDF

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US20040115623A1
US20040115623A1 US10/692,556 US69255603A US2004115623A1 US 20040115623 A1 US20040115623 A1 US 20040115623A1 US 69255603 A US69255603 A US 69255603A US 2004115623 A1 US2004115623 A1 US 2004115623A1
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Fred Homa
Michael Wathen
Todd Hopkins
Darrell Thomsen
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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
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/18—Testing for antimicrobial activity of a material
    • 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
    • A61P31/20—Antivirals for DNA viruses
    • A61P31/22—Antivirals for DNA viruses for herpes viruses
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
    • G01N2333/01—DNA viruses
    • G01N2333/03—Herpetoviridae, e.g. pseudorabies virus

Definitions

  • the present invention relates to a method for selecting an anti-herpes viral compound and a method for selectively inhibiting herpes viruses in a human host in need of such treatment.
  • the herpesviruses comprise a large family of double stranded DNA viruses. Eight of the herpes viruses, herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), varicella zoster virus (VZV), human cytomegalovirus (HCMV), Epstein-Barr virus (EBV), and human herpes viruses 6, 7, and 8 (HHV-6, HHV-7, and HHV-8), have been shown to infect humans. Several of these viruses are important human pathogens.
  • HSV-1 is estimated to affect 100 million people in the U.S. Primary infection of HSV-1 usually occurs between the ages of one and four. Cold sores, the visible symptom, typically appear at a later age, with 20-45% of the population over the age of fifteen affected (Whitley, Clin. Intect. Dis., 26:541-555, 1998).
  • Genital herpes (HSV-2) is the second most common sexually transmitted disease, with approximately 22% of the U.S population infected with this virus (Fleming 1997).
  • VZV is the causative agent of chicken pox upon primary infection and can recur in adults as zoster.
  • EBV results in approximately two million cases of infectious mononucleosis in the U.S. each year. It can also cause lymphomas in immunocompromised patients and has been associated with Burkitt's lymphoma, nasopharyngeal carcinoma, and Hodgkins disease.
  • HHV-6 is the causitive agent of roseola and may be associated with multiple sclerosis and chronic fatigue syndrome. HHV-7 disease association is unclear, but it may be involved in some cases of roseola. HHV-8 has been associated with Karposi's sarcoma, body cavity based lymphomas, and multiple myeloma.
  • viruses are capable of residing in a latent state within the host. Reactivation of latent virus results from response to environmental stimuli (ex. UV exposure, stress, etc.). Infections or recurrence can be life threatening in immunocompromised patients such as AIDS or transplant patients where HCMV can result in retinitis, pneumonia, and gastrointestinal disease.
  • the increased immunocompromised population has created an unmet medical need for antivirals against herpesviruses because current therapies do not have a sufficiently broad spectrum against this family of viruses and/or they have limited utility due to toxicity.
  • the present invention provides a method for selectively inhibiting herpesviruses DNA polymerase with compounds that have broad spectrum activity. The method offers a distinct advantage in the treatment of patients in need, particularly immunocompromised patients at risk of infection or reactivation by many members of the herpesvirus family.
  • the present invention provides a method of selecting compounds that inhibit herpes viruses comprising:
  • step d) selecting the compound of interest wherein the IC 50 of step b is at least 3 times greater than the IC 50 of step a.
  • step a and step b are interchangeable.
  • the present invention further provides a method of selecting compounds that inhibit herpes viruses comprising:
  • step b selecting the compound of interest wherein the IC 50 of step b is at least 3 times greater than the IC 50 of step a.
  • step a and step b are interchangeable.
  • the present invention further provides a method of selecting compounds that inhibit herpes viruses comprising:
  • step d) selecting the compound of interest wherein the IC 50 of step b is at least 3 times greater than the IC 50 of step a.
  • step a and step b are interchangeable.
  • the present invention further provides a method of selecting compounds that inhibit herpes viruses comprising:
  • step b selecting the compound of interest wherein the IC 50 of step b is at least 3 times greater than the IC 50 of step a.
  • step a and step b are interchangeable.
  • the present invention further provides a method for selectively treating diseases caused by herpes viruses in a human host comprising administering a compound to a human in need of such treatment wherein said compound inhibits herpes viruses by interaction with the binding domain in the viral DNA polymerase.
  • the present invention further provides method for selectively inhibiting herpes viruses in a human host comprising administering a compound to a human in need of such treatment wherein IC 50 of the compound that inhibits a binding domain mutant herpes virus is at lease 3 times greater than IC 50 of the compound that inhibits a wild type herpes virus which is the same strain as the mutant herpes virus.
  • the present invention further provides a compound for treating herpesviral infections in a human host wherein IC 50 of the compound that inhibits a binding domain mutant herpes virus is at lease 5 times greater than IC 50 of the compound that inhibits a wild type herpes virus which is the same strain as the mutant herpes virus.
  • the present invention further provides a compound for treating herpesviral infections in a human host wherein said compound inhibits the herpesvirus by interacting with the binding domain in the viral DNA polymerase.
  • the present invention further provides a compound for the inhibiting of herpesvirus DNA polymerases wherein serial passage of a wild type herpes virus in the presence of said compound results in a change of the wild type HSV-1 polymerase at amino acid 823 from valine to alanine.
  • the present invention further provides a compound for inhibiting herpesvirus DNA polymerases wherein serial passage of a wild type herpes virus in the presence of said compound results a change of the wild type HCMV polymerase at amino acid 823 from valine to alanine and at amino acid 824 from valine to leucine.
  • the present invention further provides a mutant herpesvirus DNA molecule having a nucleotide sequence selected from a group consisting of SEQ.ID.NO. 1; SEQ.ID.NO. 3; SEQ.ID.NO. 5; SEQ.ID.NO. 7; SEQ.ID.NO. 9; and SEQ.ID.NO. 11.
  • the present invention further provides a mutant herpesvirus polymerase amino acid molecule having an amino acid sequence selected from a group consisting of SEQ.ID.NO. 2; SEQ.ID.NO. 4; SEQ.ID.NO. 6; SEQ.ID.NO. 8; SEQ.ID.NO. 10 and SEQ.ID.NO. 12.
  • FIG. 1 examples of 4-oxo-DHQ and 4-oxo-DHTP compounds.
  • FIG. 2 Herpesvirus' polymerases amino acid conserved region.
  • FIG. 3 Recovered virus after serial passage of HSV-1 in presence of 20 ⁇ M of compound No. 17.
  • FIG. 4 Comparision of Wild HSV-1 and HSV-2 herpesvirus DNA polymerase amino acid sequences alligned by amino acid homology. (Seq. No: 14-19)
  • FIG. 5 Meat Herpes Virus DNA and amino acid sequence list. (Seq. No: 1-12)
  • FIG. 6 Wild HCMV herpesvirus DNA polymerases amino acid sequence. (Seq. No 13)
  • a key enzyme in the replication of all herpesviruses is the virus-coded DNA polymerase.
  • Most of the currently available anti-herpes drugs target the viral DNA polymerase.
  • Drugs such as Foscarnet acts by direct inhibition of the viral polymerase.
  • These drugs are non-nucleoside inhibitors of herpesvirus DNA polymerases.
  • Others such as the nucleoside analogs, Acyclovir, Penciclovir and Ganciclovir must first be phosphorylated to the monophosphate forms by virus encoded kinases and, further phosphorylated to triphosphate by cellular enzymes before they are active inhibitors.
  • the triphosphate forms of these nucleoside analogs inhibit polymerases by competing with the binding of natural triphosphates and their subsequent insertion into growing DNA strands.
  • These drugs are known as nucleoside inhibitors of herpesvirus DNA polymerases.
  • One of the limitations of the currently available drugs is that they are active against only a few of the eight human herpesviruses.
  • Acyclovir and Penciclovir inhibit HSV and VZV replication but have poor activity against CMV.
  • ring A is a saturated or unsaturated fused double or triple heterocyclic ring having 1, 2, 3 or 4 heteroatoms selected from group consisting of oxygen, sulfur, or nitrogen; and wherein R and X are the appropriated substitutents, respectively.
  • FIG. 1 Examples of 4-HQ compounds, 4-oxo-DHQ compounds and 4-oxo-DHTP compounds are illustrated in FIG. 1.
  • Antiviral activity of these examples are shown in Table 1 below. As shown in Table 1, these compounds inhibit HSV-1 and HSV-2 as well or better than the current commercially available drug Acyclovir. TABLE 1 Antiviral Activity of 4-oxo DHQ/4-oxo DTHP against HSV-1 and HSV-2 Compound IC 50 (uM) virus 1 2 3 4 5 ACV HSV-1 KOS 2.0 3.8 3.2 3.2 3.3 3.6 HSV-1 F 2.5 2.3 2.2 2.1 1.3 HSV-1 DJL 2.5 2.6 1.8 2.2 2.7 1.8 HSV-1 Patton ND 5.3 7.7 4.3 10 9.3 HSV-2 MS 2.0 2.5 2.8 2.5 2.5 10 HSV-2 35D ND 5.4 5.0 3.2 8.1 6.0 HSV-2 186 2.0 2.3 3.2 2.3 4.2 >10
  • the HSV-1, HSV-2 and HCMV mutants are also found to be resistant to other non-nucleoside inhibitors such as the 4-oxo-DHTP and similar compounds.
  • the binding domain mutants e.g. HSV-1 V823A, HSV-2-MS V826A, HSV-2-186 V828A, and HCMV V823A/V824L mutants
  • the non-nucleoside inhibitor such as Foscarnet
  • this invention provides a method for selecting compounds to treat individuals such as immunocompromised patients who are afflicted with multple herpesvirus infections.
  • wild-type refers to a gene or gene product which has the characteristics of that gene or gene product when isolated from a naturally occurring source.
  • a wild-type gene is that which is most frequently observed in a population and is thus arbitrarily designated the “normal” or “wild-type” form of the gene.
  • mutant refers to a gene or gene product which displays modifications in sequence and or functional properties (i.e., altered characteristics) when compared to the wild-type gene or gene product. It is noted that naturally-occurring mutants can be isolated, these are identified by the fact that they have altered characteristics when compared to the wild-type gene or gene product.
  • IC 50 refers to concentration of a drug that inhibits virus growth by 50%.
  • Wild type HCMV is listed in SEQ. ID. NO. 13.
  • Iudr refers to antiviral drug Iododeoxyuridine.
  • Bvdu refers to antiviral drug Bromovinyldeoxyuridine.
  • ACV antiviral drug Acyclovir
  • AraC refers to antiviral drug Arabinosylcytidine.
  • AraT refers to antiviral drug Arabinosylthymine.
  • AraA refers to antiviral drug Arabinosyladenine.
  • GCV antiviral drug Ganciclovir
  • CDV refers to antiviral drug Cidofovir.
  • PFA antiviral drug Foscarnet
  • binding domain refers to a conserved region in herpesvirus DNA polymerases.
  • the herpesvirus DNA polymerases have seven (7) conserved regions.
  • the binding domain is within the thrid conserved region (see FIG. 2).
  • the binding domain contacts with an inhibitor, at least one amino acid in the binding domain mutates and provides the resistance.
  • the binding domain is at an amino acid sequence position 818-829 of the HSV-1 DNA polymerase or the homologous region in other herpes virus DNA polymerases (see FIG. 2).
  • a binding domain mutant herpes virus refers to a herpes virus containing a binding domain mutation.
  • the binding domain in HSV-1 strains, KOS, F, DJL and Patton are at amino acid sequence position 823.
  • the binding domain in HSV-2 MS-M1 strain is at amino acid sequence position 826.
  • the binding domain in HSV-2 186 strain is at amino acid sequence position 828.
  • the binding domain in HCMV AD 169 strains is at amino acid sequence position 823-824.
  • XxxxY refers to an amino acid sequence position xxx, a single amino acid X in wild type is changed to an amino acid Y.
  • V823A refers to an amino acid sequence position 823, a Valine found in wild type is changed to alanine in mutant strain.
  • V824L refers to an amino acid sequence position 824, a Valine found in wild type is changed to Leucine in mutant strain.
  • V826A refers to an amino acid sequence position 826, a Valine found in wild type is change to alanine in mutant strain.
  • V828A refers to an amino acid sequence position 828, a Valine found in wild type is change to alanine in mutant strain.
  • African green monkey kidney cells Vero
  • human foreskin fibroblast cells HFF
  • herpes viruses can be obtained from the American Type Culture Collection (ATCC).
  • Media is defined as Dulbecco's modified Eagle media (DMEM) containing 10% fetal bovine serum (FBS) and supplemented with antibiotics.
  • FBS fetal bovine serum
  • Cells are maintained in media at 37° C. in a humidified atmosphere of 5% CO 2 .
  • HSV-1 strains F, Patton and DJL, HSV-2 strains MS, 35D and 186, and HCMV strain AD169 are used in these studies.
  • Strain DJL is a clinical isolate of HSV-1 isolated in our lab from a primary oral lesion.
  • HSV-1 and HSV-2 stocks are grown in Vero cells.
  • HCMV stocks are grown in HFF cells.
  • Approximately 1 ml of media containing sufficient virus to infect approximately 0.1% to 1% of the cells (multiplicity of infection of 0.001 to 0.01 PFU/cell) is added to a T-150 cell culture flask containing a confluent monolayer of cells. The cells are incubated at 37° C. for approximately 1 hour.
  • Approximately 50 ml of media is then added to the flask and the cells are incubated at 37° C. until viral cytopathic effect (cpe) is apparent in 100% of the cells.
  • the flask is then placed at ⁇ 80° C. for at least 30 min.
  • the flask containing frozen media and cells is placed in a 37° C. water bath until the media is thawed. This process disrupts the cells and releases virus into the media. 1 ml aliquots of media containing virus are dispensed into tubes and stored at ⁇ 80° C. These aliquots of media containing virus are referred to as virus stocks.
  • Titrating Virus Stocks Aliquots of virus are thawed at 37° C. and serially diluted (10 fold dilutions) in media. 0.1 ml of each dilution of virus is placed in a single well of 24-well cell culture dish containing a confluent monolayer of cells (Vero cells for HSV-1 and HSV-2, HFF cells for HCMV) and incubated at 37° C. for 1 h. The virus innoculum is then removed and 1 ml of media containing 0.8% carboxymethylcellulose (CMC) is added to each well of the dish. The dish is incubated at 37° C.
  • CMC carboxymethylcellulose
  • HSV-1 and HSV-2 6-9 days
  • HCMV 6-9 days
  • Plaques can be observed and counted microscopically or by staining the cells with 0.1% crystal violet in 20% ethanol.
  • the virus titer which is expressed as plaque forming units (PFU) per ml is obtained by counting the plaques in a well and correcting for the dilution of the viral innoculum.
  • Plaque Reduction Assays Antiviral activity of compounds against herpesviruses such as HSV-1, HSV-2, or HCMV can be measured using plaque reduction assays. 0.1 ml of media containing approximately 50 PFU of virus is added to each well of a 24-well cell culture dish containing a confluent monolayer of cells (Vero cells for HSV-1 and HSV-2, HFF cells for HCMV). Compounds are dissolved in 100% DMSO and diluted in 100% DMSO as 200 ⁇ stocks of the desired final drug concentration. Typically 5-6 two-fold dilutions are prepared for each compound. Dilutions of compounds are then added to media containing 0.8% CMC resulting in a final 1 ⁇ drug concentration.
  • virus innoculum is removed and 1 ml of media containing 0.8% CMC and the various concentrations of compound is added to each well of the dish.
  • the dish is incubated at 37° C. for approximately 2-3 days (HSV-1 and HSV-2) or 6-9 days (HCMV) to allow sufficient growth of virus to form plaques in the cell monolayer. Plaques can be observed and counted microscopically or by staining the cells with 0.1% crystal violet in 20% ethanol. Virus inhibition is determined for each drug concentration by comparing the number of plaques in drug-containing wells to control wells that did not contain drug.
  • Antiviral activity of a compound is expressed as the concentration of compound predicted to reduce the number of plaques in a well by 50% (IC 50 ).
  • the IC 50 values are calculated by plotting the percent inhibition vs. concentration of compound using EXCEL software for linear regression.
  • Vero cells are plated out at a density of 3.5 ⁇ 10 5 cells per well in a six well tissue culture plate.
  • Cells are infected with HSV-1 KOS at a multiplicity of infection (moi) of 0.1 pfu/cell and 1 h post infection the cells are overlayed with 3 ml media containing 20 uM of a 4-oxo-DHQ.
  • Cultures are incubated for 20 h at 37° C., freeze/thawed to release cell-associated virus, and 0.1 ml of culture is used to infect a new monolayer of Vero cells (one passage). Serial passage is repeated seven times in the presence of 20 uM drug. Virus isolates are then plaque purified three times prior to preparation of stocks.
  • Virus recovered from each passage in the presence of compound No. 17 is shown in FIG. 3.
  • 4-oxo-DHQ resistant HSV-1 and HSV-2 may also be selected by the marker transfer method described below using wild-type HSV DNA and the corresponding mutant HSV polymerase gene.
  • a plasmid containing the wild-type HCMV polymerase gene is modified to contain the V823A or V823A and V824L mutations using a site-directed mutagenesis Kit (Stratagene Corp.) and following the manufactures's protocol.
  • HFF cells are plated into T25 tissue culture flasks to achieve 80% confluency at the time of the transfection.
  • Wild type HCMV AD169 DNA and plasmid DNA containing the mutant HCMV polymerase gene are mixed at a ratio of 1:2 (2 ug of viral DNA to 4 ug of plasmid DNA). DNA's are transfected using superfect transfection reagent according to methods recommended by the manufacturer (Quiagen Inc.).
  • Cells are harvested five days posttransfection, freeze-thawed to release virus and half of the sample is used to infect 1FF cell monolayers. Cells are overlayed with media containing 20 uM 4-oxo-DHQ compound 2 (see FIG. 1). Serial passage is repeated seven times in the presence of 20 uM compound 2 and virus isolates are then plaque purified three times prior to preparation of viral stock.
  • HSV DNA is purified from the cytoplasm of infected Vero cells.
  • Vero cells (50% confluent) are infected at an multiplicity of 0.01 PFU/cell.
  • infected cells 100% cpe
  • the pelleted cells are resuspended in TE buffer and placed on ice for 15 minutes.
  • NP-40 is then added to a final concentration of 0.2% and incubated on ice for a further 15 minutes.
  • the cells are centrifuged at 2000 rpm for 10 minutes in a Beckman GS-6R centrifuge.
  • HCMV DNA is isolated by infecting HFF cells (25% confluency) with HCMV at an multiplicity of 0.1 PFU/cell. Cells and media are harvested 5-7 days postinfection (100% cpe) and subjected to low speed centrifugation to remove intact cells and cell debris followed by a high speed spin to pellet virus particles (2500 rpm's in a Beckman SW28 rotor for 1 hour).
  • HSV-1, HSV-2 or HCMV viral DNA's are sequenced directly using an ABI377 fluorescence sequencer (Perkin Elmer Applied Biosystems, Foster City, Calif.) and the ABI BigDye PRISMTM dRhodamine Terminator Cycle Sequencing Ready Reaction Kit with AmpliTaq FSTM DNA polymerase (PE Applied Biosystems). Each cycle sequencing reaction contained about 1.0 ug of purified viral DNA. Cycle-sequencing is performed using an initial denaturation at 98° C. for 1 min, followed by 50 cycles: 98° C. for 30 sec, annealing at 50° C. for 30 sec, and extension at 60° C. for 4 min.
  • sequence chromatogram data files from the ABI377 are analyzed with the computer program Sequencher (Gene Codes, Ann Arbor, Mich.), for assembly of sequence fragments and correction of ambiguous base calls. Generally sequence reads of 600-700 bp are obtained. Potential sequencing errors are minimized by obtaining sequence information from both DNA strands and by re-sequencing difficult areas using primers at different locations until all sequencing ambiguities are removed.
  • the entire coding region of the polymerase genes from both the parent strains and the resistant viruses are sequenced.
  • the DNA sequencing is done using viral DNA as the template thus avoiding cloning of the polymerase genes.
  • the amino acid sequence of the DNA polymerases of HSV-1 KOS, F, Patton and DJL and HSV-2 MS and 186 are compared in FIG. 4. Amino acids that are identical for the six polymerases are shaded in black while regions where amino acid differences are found are shaded in gray.
  • the amino acid sequence of the four HSV-1 polymerases are essentially identical with only a few minor changes noted between the different HSV-1 strains. The majority of amino acid changes are found when the sequences of the HSV-1 and HSV-2 polymerases are compared.
  • a panel of viruses consisting of four strains of HSV-1 (KOS, F, DJL, Patton) and three strains of HSV-2 (MS, 35D, 186) are tested in a plaque reduction assay against four different 4-oxo-DHQ compounds (# 1, 2, 4, 5 as shown in FIG. 1), and one 4-oxo-DHTP compound (# 3 as shown in FIG. 1) and against Acyclovir.
  • the six drugs inhibited replication of the seven virus strains with IC 50 values ranging from 2-10 ⁇ M (Table 1).
  • HSV-1 strains KOS, F, and DJL along with HSV-2 strains 186 and MS are serially passaged in the presence of 20 uM compound 1.
  • 4-oxo-DHQ resistant virus from each strain are plaque purified three times and high-titer stocks are made. All of the resistant HSV mutants grew to high titers in Vero cells, indicating that the mutations in the resistant isolates did not significantly impair their growth.
  • the mutants selected with 4-oxo-DHQ compound 1 exhibited >10 fold increase in IC 50 when tested in a plaque reduction assay against 4-oxo-DHQ compound 1 Data are shown in Table 2.
  • virus (strain AD169) is serially passaged in the presence of 20 uM a 4-oxo-DHQ. Although we could readily select for HSV mutants using this procedure we failed to isolate an HCMV mutant, even when the virus is passaged at low drug concentrations ( ⁇ 5 uM).
  • V823A alone resulted in a 3.5-fold increase in the IC 50 while the polymerase with the double amino acid change had nearly 10-fold increase in the IC 50 .
  • HCMV resistant mutant marker rescue experiments are done. Plasmids containing the mutant polymerase genes are transfected into HFF cells along with wild type HCMV AD169 DNA. The resulting virus is then serially passaged in the presence of 20 uM compound 1 (see FIG. 1). A 4-oxo-DHQ resistant virus is isolated from marker rescue studies done with the HCMV polymerase gene containing mutations that result in the V823A, V824L amino acid changes, but not with the gene containing V823A change alone.
  • the mutant selected with compound 1 (HCMV AD169-M1) exhibited ⁇ 7-fold increase in IC 50 when tested in a plaque reduction assay compared to Ganciclovir and cidofovir which has a ⁇ 2-fold change in sensitivity (Table 4).
  • TABLE 4 Plaque reduction assay of 4-oxo-DHQ resistant HCMV* HCMV AD169 HCMV AD169-M1 Drug IC 50 ( ⁇ M) IC 50 ( ⁇ M)
  • Compound 1 0.7 4.7 Ganciclovir 0.9 1.0 Cidofovir 0.3 0.6
  • the activity of these compounds against the mutants is compared to their activity against the wild type strains that are used to isolate the HSV and HCMV mutants.
  • all of the drugs are found to inhibit the wild type virus with IC 50 values ranging from ⁇ 0.1 uM to 30 uM.
  • these drugs are tested against the resistant viruses they are found to have IC 50 values 5 to 10 fold higher then the parent virus. There is little if any difference in the IC 50 values of the nucleoside compounds and the non-nucleoside PFA between the wild type and mutant HSV-1, HSV-2, and HCMV viruses.
  • Antiviral compounds identified by the present invention can conveniently be administered in a pharmaceutical composition containing the compound in combination with a suitable excipient, the composition being useful in combating viral infections.
  • Pharmaceutical compositions containing a compound appropriate for antiviral use are prepared by methods and contain excipients which are well known in the art. A generally recognized compendium of such methods and ingredients is Remington's Pharmaceutical Sciences by E. W. Martin (Mark Publ. Co., 15th Ed., 1975).
  • Antiviral compounds identified by the present invention and their compositions can by administered parenterally (for example, by intravenous, intraperitoneal or intramuscular injection), topically, orally, or rectally, depending on whether the preparation is used to treat internal or external viral infections.
  • the active compound may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like.
  • Such compositions and preparations should contain at least 0.1% of active compound.
  • the percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 2 to about 60% of the weight of a given unit dosage form.
  • the amount of active compound in such therapeutically useful compositions is such that an effective dosage level will be obtained.
  • the tablets, troches, pills, capsules, and the like may also contain the following: binders such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, fructose, lactose or aspartame or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring may be added.
  • a liquid carrier such as a vegetable oil or a polyethylene glycol.
  • any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed.
  • the active compound may be incorporated into sustained-release preparations and devices.
  • Antiviral compounds identified by the present invention and their compositions can also be administered intravenously or intraperitoneally by infusion or injection.
  • Solutions of the active compound or its salts can be prepared in water, optionally mixed with a nontoxic surfactant.
  • Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
  • compositions suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes.
  • the liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof.
  • the proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants.
  • the prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.
  • Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilization.
  • the preferred methods of preparation are vacuum drying and the freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solutions.
  • the present compounds may be applied in pure form, i.e., when they are liquids. However, it will generally be desirable to administer them to the skin as compositions or formulations, in combination with a dermatologically acceptable carrier, which may be a solid or a liquid.
  • Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina and the like.
  • Useful liquid carriers include water, alcohols or glycols or water-alcohol/glycol blends, in which the present compounds can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants.
  • Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize the properties for a given use.
  • the resultant liquid compositions can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using pump-type or aerosol sprayers.
  • Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses or modified mineral materials can also be employed with liquid carriers to form spreadable pastes, gels, ointments, soaps, and the like, for application directly to the skin of the user.
  • Examples of useful dermatological compositions which can be used to deliver the compounds of formula I to the skin are known to the art; for example, see Jacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S. Pat. No. 4,992,478), Smith et al. (U.S. Pat. No. 4,559,157) and Wortzman (U.S. Pat. No. 4,820,508).
  • Useful dosages of the compounds of formula I can be determined by comparing their in vitro activity, and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art; for example, see U.S. Pat. No. 4,938,949.
  • the compound is conveniently administered in unit dosage form; for example, containing 5 to 1000 mg, conveniently 10 to 750 mg, most conveniently, 50 to 500 mg of active ingredient per unit dosage form.
  • the desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day.
  • the sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations; such as multiple inhalations from an insufflator or by application of a plurality of drops into the eye.
  • compositions can be administered orally or parenterally at dose levels, calculated as the free base, of about 0.1 to 300 mg/kg, preferably 1.0 to 30 mg/kg of mammal body weight, and can be used in man in a unit dosage form, administered one to four times daily in the amount of 1 to 1000 mg per unit dose.
  • the compounds are presented in aqueous solution in a concentration of from about 0.1 to about 10%, more preferably about 0.1 to about 7%.
  • the solution may contain other ingredients, such as emulsifiers, antioxidants or buffers.
  • the concentration of the compound(s) of formula I in a liquid composition will be from about 0.1-25 wt-%, preferably from about 0.5-10 wt-%.
  • concentration in a semi-solid or solid composition such as a gel or a powder will be about 0.1-5 wt-%, preferably about 0.5-2.5 wt-%.
  • the antiviral activity of a compound of the invention can be determined using pharmacological models which are well known to the art, or using Test A described below.
  • the compounds of formula (I) and pharmaceutically acceptable salts thereof are useful as antiviral agents. Thus, they are useful to combat viral infections in animals, including man.
  • the compounds are generally active against herpes viruses, and are particularly useful against the varicella zoster virus, the Epstein-Barr virus, the herpes simplex virus, the human herpes virus type 8 (HHV-8) and the cytomegalovirus (CMV).

Abstract

The present invention relates to a method for selecting an anti-herpes viral compound and a method for selectively inhibiting herpesvrus in a human host in need of such treatment.
The present invention relates to a method for selecting an anti-herpes viral compound and a method for selectively inhibiting herpesvrus in a human host in need of such treatment.

Description

    CROSS REFERENCE
  • This application claims the benefit of the following provisional applications: U.S. Serial No. 60/218,118, filed Jul. 13, 2000, Serial No. 60/283,880, filed Apr. 13, 2001 under 35 USC 119(e)(i).[0001]
    • FIELD OF THE INVENTION
  • The present invention relates to a method for selecting an anti-herpes viral compound and a method for selectively inhibiting herpes viruses in a human host in need of such treatment. [0002]
    • BACKGROUND OF THE INVENTION
  • The herpesviruses comprise a large family of double stranded DNA viruses. Eight of the herpes viruses, herpes [0003] simplex virus types 1 and 2 (HSV-1 and HSV-2), varicella zoster virus (VZV), human cytomegalovirus (HCMV), Epstein-Barr virus (EBV), and human herpes viruses 6, 7, and 8 (HHV-6, HHV-7, and HHV-8), have been shown to infect humans. Several of these viruses are important human pathogens.
  • HSV-1 is estimated to affect 100 million people in the U.S. Primary infection of HSV-1 usually occurs between the ages of one and four. Cold sores, the visible symptom, typically appear at a later age, with 20-45% of the population over the age of fifteen affected (Whitley, Clin. Intect. Dis., 26:541-555, 1998). [0004]
  • Genital herpes (HSV-2) is the second most common sexually transmitted disease, with approximately 22% of the U.S population infected with this virus (Fleming 1997). [0005]
  • VZV is the causative agent of chicken pox upon primary infection and can recur in adults as zoster. [0006]
  • EBV results in approximately two million cases of infectious mononucleosis in the U.S. each year. It can also cause lymphomas in immunocompromised patients and has been associated with Burkitt's lymphoma, nasopharyngeal carcinoma, and Hodgkins disease. [0007]
  • Infection with HCMV often occurs during childhood and is typically asymptomatic except in immunocompriomised patients where it causes significant morbidity and mortality. [0008]
  • HHV-6 is the causitive agent of roseola and may be associated with multiple sclerosis and chronic fatigue syndrome. HHV-7 disease association is unclear, but it may be involved in some cases of roseola. HHV-8 has been associated with Karposi's sarcoma, body cavity based lymphomas, and multiple myeloma. [0009]
  • These viruses are capable of residing in a latent state within the host. Reactivation of latent virus results from response to environmental stimuli (ex. UV exposure, stress, etc.). Infections or recurrence can be life threatening in immunocompromised patients such as AIDS or transplant patients where HCMV can result in retinitis, pneumonia, and gastrointestinal disease. [0010]
  • The increased immunocompromised population has created an unmet medical need for antivirals against herpesviruses because current therapies do not have a sufficiently broad spectrum against this family of viruses and/or they have limited utility due to toxicity. The present invention provides a method for selectively inhibiting herpesviruses DNA polymerase with compounds that have broad spectrum activity. The method offers a distinct advantage in the treatment of patients in need, particularly immunocompromised patients at risk of infection or reactivation by many members of the herpesvirus family. [0011]
    • SUMMARY OF THE INVENTION
  • The present invention provides a method of selecting compounds that inhibit herpes viruses comprising: [0012]
  • a) measuring IC[0013] 50 of a compound of interest that inhibits a wild type herpes virus,
  • b) measuring IC[0014] 50 of the same compound that inhibits a binding domain mutant herpes virus which is the same strain of the wild type herpes virus,
  • c) comparing IC[0015] 50 of step a with IC50 of step b; and
  • d) selecting the compound of interest wherein the IC[0016] 50 of step b is at least 3 times greater than the IC50 of step a.
  • In above method, the order of step a and step b are interchangeable. [0017]
  • The present invention further provides a method of selecting compounds that inhibit herpes viruses comprising: [0018]
  • a) measuring IC[0019] 50 of a compound of interest that inhibits a wild type HSV-1,
  • b) measuring IC[0020] 50 of the same compound that inhibits a binding domain mutant HSV-1 which is the same strain of the wild type herpes virus,
  • c) comparing IC[0021] 50 of step a with IC50 of step b; and
  • d) selecting the compound of interest wherein the IC[0022] 50 of step b is at least 3 times greater than the IC50 of step a.
  • In above method, the order of step a and step b are interchangeable. [0023]
  • The present invention further provides a method of selecting compounds that inhibit herpes viruses comprising: [0024]
  • a) measuring IC[0025] 50 of a compound of interest that inhibits a wild type HSV-2,
  • b) measuring IC[0026] 50 of the same compound that inhibits a binding domain mutant HSV-2 which is the same strain of the wild type herpes virus,
  • c) comparing IC[0027] 50 of step a with IC50 of step b; and
  • d) selecting the compound of interest wherein the IC[0028] 50 of step b is at least 3 times greater than the IC50 of step a.
  • In above method, the order of step a and step b are interchangeable. [0029]
  • The present invention further provides a method of selecting compounds that inhibit herpes viruses comprising: [0030]
  • a) measuring IC[0031] 50 of a compound of interest that inhibits a wild type HCMV,
  • b) measuring IC[0032] 50 of the same compound that inhibits a binding domain mutant HCMV which is the same strain of the wild type herpes virus,
  • c) comparing IC[0033] 50 of step a with IC50 of step b; and
  • d) selecting the compound of interest wherein the IC[0034] 50 of step b is at least 3 times greater than the IC50 of step a.
  • In above method, the order of step a and step b are interchangeable. [0035]
  • The present invention further provides a method for selectively treating diseases caused by herpes viruses in a human host comprising administering a compound to a human in need of such treatment wherein said compound inhibits herpes viruses by interaction with the binding domain in the viral DNA polymerase. [0036]
  • The present invention further provides method for selectively inhibiting herpes viruses in a human host comprising administering a compound to a human in need of such treatment wherein IC[0037] 50 of the compound that inhibits a binding domain mutant herpes virus is at lease 3 times greater than IC50 of the compound that inhibits a wild type herpes virus which is the same strain as the mutant herpes virus.
  • The present invention further provides a compound for treating herpesviral infections in a human host wherein IC[0038] 50 of the compound that inhibits a binding domain mutant herpes virus is at lease 5 times greater than IC50 of the compound that inhibits a wild type herpes virus which is the same strain as the mutant herpes virus.
  • The present invention further provides a compound for treating herpesviral infections in a human host wherein said compound inhibits the herpesvirus by interacting with the binding domain in the viral DNA polymerase. [0039]
  • The present invention further provides a compound for the inhibiting of herpesvirus DNA polymerases wherein serial passage of a wild type herpes virus in the presence of said compound results in a change of the wild type HSV-1 polymerase at amino acid 823 from valine to alanine. [0040]
  • The present invention further provides a compound for inhibiting herpesvirus DNA polymerases wherein serial passage of a wild type herpes virus in the presence of said compound results a change of the wild type HCMV polymerase at amino acid 823 from valine to alanine and at amino acid 824 from valine to leucine. [0041]
  • The present invention further provides a mutant herpesvirus DNA molecule having a nucleotide sequence selected from a group consisting of SEQ.ID.NO. 1; SEQ.ID.NO. 3; SEQ.ID.NO. 5; SEQ.ID.NO. 7; SEQ.ID.NO. 9; and SEQ.ID.NO. 11. [0042]
  • The present invention further provides a mutant herpesvirus polymerase amino acid molecule having an amino acid sequence selected from a group consisting of SEQ.ID.NO. 2; SEQ.ID.NO. 4; SEQ.ID.NO. 6; SEQ.ID.NO. 8; SEQ.ID.NO. 10 and SEQ.ID.NO. 12.[0043]
    • BRIEF DESCRIPTION OF THE DRAWINGS
  • FIG. 1—examples of 4-oxo-DHQ and 4-oxo-DHTP compounds. [0044]
  • FIG. 2—Herpesvirus' polymerases amino acid conserved region. [0045]
  • FIG. 3—Recovered virus after serial passage of HSV-1 in presence of 20 μM of compound No. 17. [0046]
  • FIG. 4—Comparision of Wild HSV-1 and HSV-2 herpesvirus DNA polymerase amino acid sequences alligned by amino acid homology. (Seq. No: 14-19) [0047]
  • FIG. 5—Mutant Herpes Virus DNA and amino acid sequence list. (Seq. No: 1-12) [0048]
  • FIG. 6—Wild HCMV herpesvirus DNA polymerases amino acid sequence. (Seq. No 13)[0049]
    • DETAILED DESCRIPTION OF THE INVENTION
  • A key enzyme in the replication of all herpesviruses is the virus-coded DNA polymerase. Most of the currently available anti-herpes drugs target the viral DNA polymerase. Drugs such as Foscarnet acts by direct inhibition of the viral polymerase. These drugs are non-nucleoside inhibitors of herpesvirus DNA polymerases. Others such as the nucleoside analogs, Acyclovir, Penciclovir and Ganciclovir must first be phosphorylated to the monophosphate forms by virus encoded kinases and, further phosphorylated to triphosphate by cellular enzymes before they are active inhibitors. The triphosphate forms of these nucleoside analogs inhibit polymerases by competing with the binding of natural triphosphates and their subsequent insertion into growing DNA strands. These drugs are known as nucleoside inhibitors of herpesvirus DNA polymerases. [0050]
  • One of the limitations of the currently available drugs is that they are active against only a few of the eight human herpesviruses. For example, Acyclovir and Penciclovir inhibit HSV and VZV replication but have poor activity against CMV. [0051]
  • In order to identify antiviral compounds that would have the potential to inhibit replication of most of the human herpesviruses, compounds are in vitro screened for inhibitors of herpesvirus DNA polymerase activity. Because portions of the amino acid sequence of the polymerases are highly conserved within the herpesvirus family it is possible to discover small molecules that inhibit herpesvirus polymerases but not cellular DNA polymerases. Using this biochemical approach, several new classes of compounds such as the 4-hydroxyquinoline derivatives (4-HQ), 4-oxo-dihydroquinoline derivatives (4-oxo-DHQ) and 4-oxo-dihydrothienopyridine derivatives (4-oxo-DHTP) were discovered as potent, non-nucleoside herpesvirus DNA polymerase inhibitors. In vitro polymerase assays and/or in vivo cell culture assays have demonstrated that these compounds inhibit HSV-1, HSV-2, HCMV, VZV, EBV, and HHV-8 replication. [0052]
  • 4-Oxo-DHQ and 4-oxo-DHTP are derivatives of formula I [0053]
    Figure US20040115623A1-20040617-C00001
  • wherein ring A is a saturated or unsaturated fused double or triple heterocyclic ring having 1, 2, 3 or 4 heteroatoms selected from group consisting of oxygen, sulfur, or nitrogen; and wherein R and X are the appropriated substitutents, respectively. [0054]
  • Examples of 4-HQ compounds, 4-oxo-DHQ compounds and 4-oxo-DHTP compounds are illustrated in FIG. 1. [0055]
  • Antiviral activity of these examples are shown in Table 1 below. As shown in Table 1, these compounds inhibit HSV-1 and HSV-2 as well or better than the current commercially available drug Acyclovir. [0056]
    TABLE 1
    Antiviral Activity of 4-oxo DHQ/4-oxo DTHP Against HSV-1
    and HSV-2
    Compound IC50 (uM)
    virus 1 2 3 4 5 ACV
    HSV-1 KOS 2.0 3.8 3.2 3.2 3.3 3.6
    HSV-1 F 2.5 2.3 2.2 2.1 2.6 1.3
    HSV-1 DJL 2.5 2.6 1.8 2.2 2.7 1.8
    HSV-1 Patton ND 5.3 7.7 4.3 10 9.3
    HSV-2 MS 2.0 2.5 2.8 2.5 2.5 10
    HSV-2 35D ND 5.4 5.0 3.2 8.1 6.0
    HSV-2 186 2.0 2.3 3.2 2.3 4.2 >10
  • It has also been discovered that point mutations within the HSV-1 polymerase gene that confer resistance to Acyclovir and other nucleoside analogs do not result in resistance to the 4-HQ, 4-oxo-DHQs or 4-oxo-DHTPs. Serial passage of wild type HSV-1 in the presence of 4-oxo-DHQ results in the isolation of mutants that are highly resistant (>20 fold increase in the IC[0057] 50) to these compounds while retaining sensitivity to nucleoside inhibitors such as Acyclovir.
  • In order to determine the mechanism of action of 4-HQ, 4-oxo-DHQ and 4-oxo-DHTP compounds against herpes viruses, mutants resistant to these compounds are isolated by serial passage of the virus in the presence of a 4-oxo-DHQ compound. Sequencing analysis of HSV-1 and HSV-2 strains resistant to the 4-oxo-DHQ identifies that HSV-1 (KOS strain) polymerase protein and its homologous HSV-2 have a conserved region (a binding domain), which is a critical contact point for these compounds. While amino acid numbering of the DNA polymerase may vary between strains of HSV-1 and HSV-2, this binding domain encompassing the HSV-1 (KOS) strain amino acid 823 is highly conserved in herpesviruses and can be identified by alligning the homologous amino acids of this domain as shown in FIG. 2. [0058]
  • In HSV-1 and HSV-2 strains resistant to the 4-oxo-DHQ and similar compounds, a change of valine to an alanine at the binding domain provides full resistance. [0059]
  • In the HSV-1 DNA polymerase, resistance is also found when a valine changes to methionine at amino acid 823 but only when accompanied by a second amino acid change. [0060]
  • Isolation of HCMV resistant to 4-oxo-DHQ's is found to be very difficult. Comparison of the amino acid sequence of the HSV polymerase (Y-G-F-T-G-[0061] V-Q-H-G) and HCMV polymerase (Y-G-F-T-G-V-V-N-G) in the region of amino acid 823 (underlined amino acid) shows that there is a second valine at position 824 in the HCMV polymerase. In vitro assay using mutant HCMV polymerases demonstrates that full resistance to the 4-oxo-DHQs requires changes at both amino acids 823 (a valine to alanine) and 824 (a valine to leucine). A HCMV polymerase gene containing V823A and V824L mutations is used in marker rescue experiments to generate a viral mutant. This mutant has an IC50 approximately 7-fold above that of wild-type HCMV.
  • The HSV-1, HSV-2 and HCMV mutants are also found to be resistant to other non-nucleoside inhibitors such as the 4-oxo-DHTP and similar compounds. However, when the binding domain mutants (e.g. HSV-1 V823A, HSV-2-MS V826A, HSV-2-186 V828A, and HCMV V823A/V824L mutants) are tested in plaque reduction assays against a series of nucleoside polymerase inhibitors and the non-nucleoside inhibitor such as Foscarnet, replication of the mutants is found to be inhibited by all of the currently marketed anti-herpes polymerase inhibitors tested. [0062]
  • These studies demonstrate that certain non-nucleosides like 4-HQ, 4-oxo-DHQ and 4-oxo-DHTP compounds bind to a different site on the herpes polymerase than the nucleoside inhibitors and Foscarnet. The valine at the binding domain is conserved in the DNA polymerases of six of the eight human herpesviruses and several animal herpesviruses, and appears to play a critical role in the antiviral activity of the 4-HQ, 4-oxo-DHQ and 4-oxo-DHTP compounds. (See FIG. 2) [0063]
  • Since mutation at the binding domain negates these non-nucleoside inhibitors' activities, compounds could be tested against wild type polymerases and the mutant polymerases to establish the probability of similar binding. We refer to this property of compounds as interaction with the binding domain. Since compounds that interact with the binding domain have exhibited broad-spectrum activity against herpesviruses, this invention provides a method for selecting compounds to treat individuals such as immunocompromised patients who are afflicted with multple herpesvirus infections. [0064]
  • Definitions [0065]
  • The term “wild-type” refers to a gene or gene product which has the characteristics of that gene or gene product when isolated from a naturally occurring source. A wild-type gene is that which is most frequently observed in a population and is thus arbitrarily designated the “normal” or “wild-type” form of the gene. [0066]
  • In contrast, the term “mutant” refers to a gene or gene product which displays modifications in sequence and or functional properties (i.e., altered characteristics) when compared to the wild-type gene or gene product. It is noted that naturally-occurring mutants can be isolated, these are identified by the fact that they have altered characteristics when compared to the wild-type gene or gene product. [0067]
  • IC[0068] 50 refers to concentration of a drug that inhibits virus growth by 50%.
  • Wild type HSV-1 and HSV-2 strains are listed in FIG. 4. [0069]
  • Wild type HCMV is listed in SEQ. ID. NO. 13. [0070]
  • The term “Iudr” refers to antiviral drug Iododeoxyuridine. [0071]
  • The term “Bvdu” refers to antiviral drug Bromovinyldeoxyuridine. [0072]
  • The term “ACV” refers to antiviral drug Acyclovir. [0073]
  • The term “AraC” refers to antiviral drug Arabinosylcytidine. [0074]
  • The term “AraT” refers to antiviral drug Arabinosylthymine. [0075]
  • The term “AraA” refers to antiviral drug Arabinosyladenine. [0076]
  • The term “GCV” refers to antiviral drug Ganciclovir. [0077]
  • The term “CDV” refers to antiviral drug Cidofovir. [0078]
  • The term “PFA” refers to antiviral drug Foscarnet. [0079]
  • The term “binding domain” refers to a conserved region in herpesvirus DNA polymerases. The herpesvirus DNA polymerases have seven (7) conserved regions. The binding domain is within the thrid conserved region (see FIG. 2). When the binding domain contacts with an inhibitor, at least one amino acid in the binding domain mutates and provides the resistance. In general, the binding domain is at an amino acid sequence position 818-829 of the HSV-1 DNA polymerase or the homologous region in other herpes virus DNA polymerases (see FIG. 2). [0080]
  • The term “a binding domain mutant herpes virus” refers to a herpes virus containing a binding domain mutation. [0081]
  • More specifically, the binding domain in HSV-1 strains, KOS, F, DJL and Patton are at amino acid sequence position 823. The binding domain in HSV-2 MS-M1 strain is at amino [0082] acid sequence position 826. The binding domain in HSV-2 186 strain is at amino acid sequence position 828. The binding domain in HCMV AD 169 strains is at amino acid sequence position 823-824.
  • The term “XxxxY” refers to an amino acid sequence position xxx, a single amino acid X in wild type is changed to an amino acid Y. [0083]
  • For example, the term “V823A” refers to an amino acid sequence position 823, a Valine found in wild type is changed to alanine in mutant strain. [0084]
  • The term “V824L” refers to an amino acid sequence position 824, a Valine found in wild type is changed to Leucine in mutant strain. [0085]
  • The term “V826A” refers to an amino [0086] acid sequence position 826, a Valine found in wild type is change to alanine in mutant strain.
  • The term “V828A” refers to an amino acid sequence position 828, a Valine found in wild type is change to alanine in mutant strain. [0087]
  • A table of amino acids and their representative abbreviations, symbols and codons is set forth below in the following Table. [0088]
    Abb- Sym-
    Amino acid rev. bol Codon(s)
    Alanine Ala A GCA GCC GCG GCU
    Cysteine Cys C UGC UGU
    Aspartic acid Asp D GAC GAU
    Glutamic acid Glu E GAA GAG
    Phenylalanine Phe F UUC UUU
    Glycine Gly G GGA GGC GGG GGU
    Histidine His H CAC CAU
    Isoleucine Ile I AUA AUC AUU
    Lysine Lys K AAA AAG
    Leucine Leu L UUA UUG CUA CUC CUG CUU
    Methionine Met M AUG
    Asparagine Asn N AAC AAU
    Proline Pro P CCA CCC CCG CCU
    Glutamine Gln Q CAA CAG
    Arginine Arg R AGA AGG CGA CGC CGG CGU
    Serine Ser S AGC AGU UCA UCC UCG UCU
    Threonine Thr T ACA ACC ACG ACU
    Valine Val V GUA GUC GUG GUU
    Tryptophan Trp W UGG
    Tyrosine Tyr Y UAC UAU
    • Materials and Methods
  • Cell and Viruses [0089]
  • African green monkey kidney cells (Vero) and human foreskin fibroblast cells (HFF) and herpes viruses can be obtained from the American Type Culture Collection (ATCC). Media is defined as Dulbecco's modified Eagle media (DMEM) containing 10% fetal bovine serum (FBS) and supplemented with antibiotics. Cells are maintained in media at 37° C. in a humidified atmosphere of 5% CO[0090] 2. HSV-1 strains F, Patton and DJL, HSV-2 strains MS, 35D and 186, and HCMV strain AD169 are used in these studies. Strain DJL is a clinical isolate of HSV-1 isolated in our lab from a primary oral lesion.
  • Measuring IC[0091] 50 of a Compound of Interest that Inhibits Heroes Viruses
  • Preparation of Virus Stocks: HSV-1 and HSV-2 stocks are grown in Vero cells. HCMV stocks are grown in HFF cells. Approximately 1 ml of media containing sufficient virus to infect approximately 0.1% to 1% of the cells (multiplicity of infection of 0.001 to 0.01 PFU/cell) is added to a T-150 cell culture flask containing a confluent monolayer of cells. The cells are incubated at 37° C. for approximately 1 hour. Approximately 50 ml of media is then added to the flask and the cells are incubated at 37° C. until viral cytopathic effect (cpe) is apparent in 100% of the cells. The flask is then placed at −80° C. for at least 30 min. The flask containing frozen media and cells is placed in a 37° C. water bath until the media is thawed. This process disrupts the cells and releases virus into the media. 1 ml aliquots of media containing virus are dispensed into tubes and stored at −80° C. These aliquots of media containing virus are referred to as virus stocks. [0092]
  • Titrating Virus Stocks: Aliquots of virus are thawed at 37° C. and serially diluted (10 fold dilutions) in media. 0.1 ml of each dilution of virus is placed in a single well of 24-well cell culture dish containing a confluent monolayer of cells (Vero cells for HSV-1 and HSV-2, HFF cells for HCMV) and incubated at 37° C. for 1 h. The virus innoculum is then removed and 1 ml of media containing 0.8% carboxymethylcellulose (CMC) is added to each well of the dish. The dish is incubated at 37° C. for approximately 2-3 days (HSV-1 and HSV-2) or 6-9 days (HCMV) to allow sufficient growth of virus to form plaques in the cell monolayer. Plaques can be observed and counted microscopically or by staining the cells with 0.1% crystal violet in 20% ethanol. The virus titer which is expressed as plaque forming units (PFU) per ml is obtained by counting the plaques in a well and correcting for the dilution of the viral innoculum. [0093]
  • Plaque Reduction Assays: Antiviral activity of compounds against herpesviruses such as HSV-1, HSV-2, or HCMV can be measured using plaque reduction assays. 0.1 ml of media containing approximately 50 PFU of virus is added to each well of a 24-well cell culture dish containing a confluent monolayer of cells (Vero cells for HSV-1 and HSV-2, HFF cells for HCMV). Compounds are dissolved in 100% DMSO and diluted in 100% DMSO as 200× stocks of the desired final drug concentration. Typically 5-6 two-fold dilutions are prepared for each compound. Dilutions of compounds are then added to media containing 0.8% CMC resulting in a final 1× drug concentration. After the virus-infected cells have incubated for 1 h at 37° C., the virus innoculum is removed and 1 ml of media containing 0.8% CMC and the various concentrations of compound is added to each well of the dish. The dish is incubated at 37° C. for approximately 2-3 days (HSV-1 and HSV-2) or 6-9 days (HCMV) to allow sufficient growth of virus to form plaques in the cell monolayer. Plaques can be observed and counted microscopically or by staining the cells with 0.1% crystal violet in 20% ethanol. Virus inhibition is determined for each drug concentration by comparing the number of plaques in drug-containing wells to control wells that did not contain drug. Antiviral activity of a compound is expressed as the concentration of compound predicted to reduce the number of plaques in a well by 50% (IC[0094] 50). The IC50 values are calculated by plotting the percent inhibition vs. concentration of compound using EXCEL software for linear regression.
  • Selection of 4-oxo-DHO Resistant HSV-1 and HSV-2 [0095]
  • Vero cells are plated out at a density of 3.5×10[0096] 5 cells per well in a six well tissue culture plate. Cells are infected with HSV-1 KOS at a multiplicity of infection (moi) of 0.1 pfu/cell and 1 h post infection the cells are overlayed with 3 ml media containing 20 uM of a 4-oxo-DHQ. Cultures are incubated for 20 h at 37° C., freeze/thawed to release cell-associated virus, and 0.1 ml of culture is used to infect a new monolayer of Vero cells (one passage). Serial passage is repeated seven times in the presence of 20 uM drug. Virus isolates are then plaque purified three times prior to preparation of stocks. Virus recovered from each passage in the presence of compound No. 17 is shown in FIG. 3. 4-oxo-DHQ resistant HSV-1 and HSV-2 may also be selected by the marker transfer method described below using wild-type HSV DNA and the corresponding mutant HSV polymerase gene.
  • Marker Transfer of a HCMV Mutation [0097]
  • A plasmid containing the wild-type HCMV polymerase gene is modified to contain the V823A or V823A and V824L mutations using a site-directed mutagenesis Kit (Stratagene Corp.) and following the manufactures's protocol. HFF cells are plated into T25 tissue culture flasks to achieve 80% confluency at the time of the transfection. Wild type HCMV AD169 DNA and plasmid DNA containing the mutant HCMV polymerase gene are mixed at a ratio of 1:2 (2 ug of viral DNA to 4 ug of plasmid DNA). DNA's are transfected using superfect transfection reagent according to methods recommended by the manufacturer (Quiagen Inc.). Cells are harvested five days posttransfection, freeze-thawed to release virus and half of the sample is used to infect 1FF cell monolayers. Cells are overlayed with media containing 20 uM 4-oxo-DHQ compound 2 (see FIG. 1). Serial passage is repeated seven times in the presence of 20 [0098] uM compound 2 and virus isolates are then plaque purified three times prior to preparation of viral stock.
  • Isolation of HSV and HCMV Viral DNA [0099]
  • HSV DNA is purified from the cytoplasm of infected Vero cells. Vero cells (50% confluent) are infected at an multiplicity of 0.01 PFU/cell. At 3-5 days postinfection infected cells (100% cpe) are harvested by centrifugation at 1000 rpm in a Beckman GS-6R centrifuge. The pelleted cells are resuspended in TE buffer and placed on ice for 15 minutes. NP-40 is then added to a final concentration of 0.2% and incubated on ice for a further 15 minutes. The cells are centrifuged at 2000 rpm for 10 minutes in a Beckman GS-6R centrifuge. The supernatant is removed and EDTA is added to a final concentration of 20 mM followed by the addition of SDS to a final concentration of 0.3% and proteinase K to a concentration of 50 ug/ml then incubated at 45 C for 2 hours. HCMV DNA is isolated by infecting HFF cells (25% confluency) with HCMV at an multiplicity of 0.1 PFU/cell. Cells and media are harvested 5-7 days postinfection (100% cpe) and subjected to low speed centrifugation to remove intact cells and cell debris followed by a high speed spin to pellet virus particles (2500 rpm's in a Beckman SW28 rotor for 1 hour). Following incubation of the HSV and HCMV samples, 1.5 volumes of saturated NaI is added to the digested extract and the refractive index is adjusted to 1.434-1.435. Ethidium bromide is added to a final concentration of 50 ug/ml. The samples are loaded into a [0100] VTI 50 centrifuge tube and spun for 24 hours at 45,000 rpm. The DNA band is harvested extracted three times with n-butanol, then dialyzed against TE buffer followed by a dialysis against 95% ethanol and a final dialysis against TE buffer.
  • DNA Sequencing [0101]
  • HSV-1, HSV-2 or HCMV viral DNA's are sequenced directly using an ABI377 fluorescence sequencer (Perkin Elmer Applied Biosystems, Foster City, Calif.) and the ABI BigDye PRISM™ dRhodamine Terminator Cycle Sequencing Ready Reaction Kit with AmpliTaq FS™ DNA polymerase (PE Applied Biosystems). Each cycle sequencing reaction contained about 1.0 ug of purified viral DNA. Cycle-sequencing is performed using an initial denaturation at 98° C. for 1 min, followed by 50 cycles: 98° C. for 30 sec, annealing at 50° C. for 30 sec, and extension at 60° C. for 4 min. Temperature cycles and times are controlled by a Perkin-Elmer 9700 thermocycler. Extension products are purified using Centriflex™ gel filtration cartridges (Edge BioSystems, Gaithersburg, Md.). Each reaction product is loaded by pipette onto the column, which is then centrifuged in a swinging bucket centrifuge (Sorvall model RT6000B table top centrifuge) at 750×g for 1.5 min at room temperature. Column-purified samples are dried under vacuum for about 40 min and then dissolved in 4 ul of a DNA loading solution (83% deionized formamide, 8.3 mM EDTA, and 1.6 mg/ml Blue Dextran). The samples are then heated to 90° C. for two min, and held at 4° C. until loading. 1.5 ul of each sample is loaded into a single well of the ABI377 sequencer. Sequence chromatogram data files from the ABI377 are analyzed with the computer program Sequencher (Gene Codes, Ann Arbor, Mich.), for assembly of sequence fragments and correction of ambiguous base calls. Generally sequence reads of 600-700 bp are obtained. Potential sequencing errors are minimized by obtaining sequence information from both DNA strands and by re-sequencing difficult areas using primers at different locations until all sequencing ambiguities are removed. [0102]
  • The entire coding region of the polymerase genes from both the parent strains and the resistant viruses are sequenced. The DNA sequencing is done using viral DNA as the template thus avoiding cloning of the polymerase genes. The amino acid sequence of the DNA polymerases of HSV-1 KOS, F, Patton and DJL and HSV-2 MS and 186 are compared in FIG. 4. Amino acids that are identical for the six polymerases are shaded in black while regions where amino acid differences are found are shaded in gray. The amino acid sequence of the four HSV-1 polymerases are essentially identical with only a few minor changes noted between the different HSV-1 strains. The majority of amino acid changes are found when the sequences of the HSV-1 and HSV-2 polymerases are compared. [0103]
  • Isolation and Characterization of HSV-1 and HSV-2 Mutants that are Resistant to the 4-oxo-DHO's and 4-oxo-DHTP Compounds [0104]
  • A panel of viruses consisting of four strains of HSV-1 (KOS, F, DJL, Patton) and three strains of HSV-2 (MS, 35D, 186) are tested in a plaque reduction assay against four different 4-oxo-DHQ compounds (# 1, 2, 4, 5 as shown in FIG. 1), and one 4-oxo-DHTP compound (# 3 as shown in FIG. 1) and against Acyclovir. The six drugs inhibited replication of the seven virus strains with IC[0105] 50 values ranging from 2-10 μM (Table 1). In order to select for 4-oxo-DHQ resistant mutants, HSV-1 strains KOS, F, and DJL along with HSV-2 strains 186 and MS are serially passaged in the presence of 20 uM compound 1. Following the seventh passage, 4-oxo-DHQ resistant virus from each strain are plaque purified three times and high-titer stocks are made. All of the resistant HSV mutants grew to high titers in Vero cells, indicating that the mutations in the resistant isolates did not significantly impair their growth. The mutants selected with 4-oxo-DHQ compound 1 exhibited >10 fold increase in IC50 when tested in a plaque reduction assay against 4-oxo-DHQ compound 1 Data are shown in Table 2.
    TABLE 2
    4-oxo-DHQ Resistant Virus of HSV-1 and HSV-2
    Compound 1 Amino Acid Change in HSV
    Virus Mutants IC50 (uM) DNA Polymerase
    HSV-1 Kos-M1 >20 -V823A
    HSV-1 F-M1 >20 -V823A
    HSV-1 DJL-M1 >20 -V823A
    HSV-2 MS-M1 >20 -V826A
    HSV-2 186-M1 >20 -V828A
  • DNA sequence analysis of the 4-oxo-DHQ resistant mutants (HSV-1 KOS-M1, HSV-1 F-M1, HSV-1 DJL-M1, HSV-2 186-M1, HSV-2 MS-M1) demonstrated that all five mutants contained a single point mutation of T to C at the binding domain resulting in a Valine to Alanine amino acid change. [0106]
  • Isolation and Characterization of A HCMV Mutant that is Resistant to the 4-oxo-DHO's and 4-oxo-DHTP Compounds [0107]
  • In order to select for a 4-oxo-DHQ HCMV resistant mutant, virus (strain AD169) is serially passaged in the presence of 20 uM a 4-oxo-DHQ. Although we could readily select for HSV mutants using this procedure we failed to isolate an HCMV mutant, even when the virus is passaged at low drug concentrations (<5 uM). Comparison of the amino acid sequence of the HSV polymerase, Y-G-F-T-G-[0108] V-Q-H-G, and HCMV polymerase, Y-G-F-T-G-V-V-N-G, in the region of amino acid 823 (underlined amino acid) showed that there is a second valine at position 824 in the HCMV polymerase. In order to determine if both valines need to be changed in order to confer resistance to the 4-oxo-DHQ's, in vitro polymerase assays are done using mutant HCMV polymerases containing either V823A or V823A plus V824L (Table 3).
    TABLE 3
    HCMV Mutant Polymerase Exhibits Resistance to 4-oxo-DHQ*
    Polymerase Compound 1 IC50 (uM)
    HCMV (wild) 4.6
    HCMV V823A 17.2
    HCMV V823A/V824L 42.9
  • The V823A alone resulted in a 3.5-fold increase in the IC[0109] 50 while the polymerase with the double amino acid change had nearly 10-fold increase in the IC50. In order to isolate an HCMV resistant mutant marker rescue experiments are done. Plasmids containing the mutant polymerase genes are transfected into HFF cells along with wild type HCMV AD169 DNA. The resulting virus is then serially passaged in the presence of 20 uM compound 1 (see FIG. 1). A 4-oxo-DHQ resistant virus is isolated from marker rescue studies done with the HCMV polymerase gene containing mutations that result in the V823A, V824L amino acid changes, but not with the gene containing V823A change alone. The mutant selected with compound 1 (HCMV AD169-M1) exhibited ˜7-fold increase in IC50 when tested in a plaque reduction assay compared to Ganciclovir and cidofovir which has a ≦2-fold change in sensitivity (Table 4).
    TABLE 4
    Plaque reduction assay of 4-oxo-DHQ resistant HCMV*
    HCMV AD169 HCMV AD169-M1
    Drug IC50 (μM) IC50 (μM)
    Compound 1 0.7 4.7
    Ganciclovir 0.9 1.0
    Cidofovir 0.3 0.6
  • [0110]
  • The entire coding region of the HCMV polymerase genes from both the parent strain and the resistant virus are sequenced. The DNA sequencing is again done using viral DNA as the template thus avoiding cloning of the polymerase genes. Comparison of the DNA sequence of the two polymerase genes demonstrated that the resistant mutant contained two point mutations that resulted in the predicted V823A, V824L amino acid changes. As with the HSV resistant viruses these results demonstrate the critical role of the region encompassing amino acid 823 for inhibition of polymerase activity by these compounds. [0111]
  • Antiviral Activity of Nucleoside and Non-Nucleoside Polymerase Inhibitors Against 4-oxo-DHO Resistant Mutants [0112]
  • In order to determine if the 4-HQ binding domain mutations alter the sensitivity of the HSV-1, HSV-2 and HCMV mutants to both non-nucleoside (4-oxo-DHQ's) and nucleoside inhibitors (e.g Acyclovir and ganciclovir) several of the mutants are tested in plaque reduction assays against a series of non-nucleoside compounds including Foscarnet (PFA), 4-HQ's 4-oxo-DHQ's and 4-oxo-DHTP's (Table 5). The mutants are also tested against a series of nucleoside inhibitors including acyclovir and ganciclovir (Table 5). The activity of these compounds against the mutants is compared to their activity against the wild type strains that are used to isolate the HSV and HCMV mutants. When tested against a number of 4-HQ's, 4-oxo-DHQ's and 4-oxo-DHTP's and other related classes of compounds all of the drugs are found to inhibit the wild type virus with IC[0113] 50 values ranging from <0.1 uM to 30 uM. When these drugs are tested against the resistant viruses they are found to have IC50 values 5 to 10 fold higher then the parent virus. There is little if any difference in the IC50 values of the nucleoside compounds and the non-nucleoside PFA between the wild type and mutant HSV-1, HSV-2, and HCMV viruses. These results demonstrate that the amino acid change in the binding domain (V823A in the HSV-1 polymerase, V826A in the HSV2-MS polymerase, V828A in the HSV2-186 polymerase, and the V823A/V824L changes in the HCMV polymerase) resulted in resistance to the 4-oxo-DHQ's and 4-oxo-DHTP's, which provides further evidence that these classes of compounds share an affinity for a region we refer to as the binding domain. In contrast, these amino acid changes did not alter the activity of these viruses to other classes of polymerase inhibitors.
    TABLE 5
    Antiviral activity of nucleoside and non-nucleoside polymerase
    inhibitors against HSV-1, HSV-2, and HCMV Isolates selected for
    4-oxo-DHQ resistance*
    Plaque Reduction Assay - IC50 (μM)
    HSV-2 HSV-2 HSV-1 HSV-1 HCMV HCMV
    Drug MS MS-M1 KOS KOS-M1 AD169 AD169-M1
     6 28.8 >50 24.6 >50 5.1 >16
     7 8.8 27.9 6.5 >50 0.3 3.4
     8 2.3 >50 5.1 >50 <0.1 1.1
     9 0.9 48.7 1.9 >50 <0.1 3.1
    10 29.2 >50 15.8 >50 1.1 >16
    11 3.0 >50 3.1 >50 0.7 3.9
    12 0.4 12.5 1.3 >50 0.2 1.1
    13 5.3 >50 5.5 <25 2.7 >16
    14 1.6 >50 28.4 >50 0.9 18.4
     2 1.3 >50 3.3 >50 0.4 4.0
     4 2.1 28.4 4.2 >50 0.6 2.1
     3 0.8 >50 4.0 >50 1.5 6.2
    15 5.9 >50 >50 >50 0.7 7.7
    Iudr 5.0 6.1 1.1 0.8 ND ND
    Bvdu 5.8 5.9 2.1 0.1 ND ND
    ACV 2.4 2.8 3.9 4.4 ND ND
    AraC 0.2 0.1 0.2 0.2 ND ND
    AraT 6.6 3.6 11.6 3.6 ND ND
    AraA 10.6 18.2 26.1 27.2 ND ND
    GCVir ND ND ND ND 0.8 0.8
    CDV ND ND ND ND 0.4 0.3
    PFA ND ND ND ND 38 <20
  • Antiviral compounds identified by the present invention can conveniently be administered in a pharmaceutical composition containing the compound in combination with a suitable excipient, the composition being useful in combating viral infections. Pharmaceutical compositions containing a compound appropriate for antiviral use are prepared by methods and contain excipients which are well known in the art. A generally recognized compendium of such methods and ingredients is Remington's Pharmaceutical Sciences by E. W. Martin (Mark Publ. Co., 15th Ed., 1975). [0114]
  • Antiviral compounds identified by the present invention and their compositions can by administered parenterally (for example, by intravenous, intraperitoneal or intramuscular injection), topically, orally, or rectally, depending on whether the preparation is used to treat internal or external viral infections. [0115]
  • For oral therapeutic administration, the active compound may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations should contain at least 0.1% of active compound. The percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 2 to about 60% of the weight of a given unit dosage form. The amount of active compound in such therapeutically useful compositions is such that an effective dosage level will be obtained. [0116]
  • The tablets, troches, pills, capsules, and the like may also contain the following: binders such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, fructose, lactose or aspartame or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier, such as a vegetable oil or a polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For instance, tablets, pills, or capsules may be coated with gelatin, wax, shellac or sugar and the like. A syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor. Of course, any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the active compound may be incorporated into sustained-release preparations and devices. [0117]
  • Antiviral compounds identified by the present invention and their compositions can also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the active compound or its salts can be prepared in water, optionally mixed with a nontoxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. [0118]
  • Pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In all cases, the ultimate dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin. [0119]
  • Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solutions. [0120]
  • For topical administration, the present compounds may be applied in pure form, i.e., when they are liquids. However, it will generally be desirable to administer them to the skin as compositions or formulations, in combination with a dermatologically acceptable carrier, which may be a solid or a liquid. [0121]
  • Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina and the like. Useful liquid carriers include water, alcohols or glycols or water-alcohol/glycol blends, in which the present compounds can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants. Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize the properties for a given use. The resultant liquid compositions can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using pump-type or aerosol sprayers. Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses or modified mineral materials can also be employed with liquid carriers to form spreadable pastes, gels, ointments, soaps, and the like, for application directly to the skin of the user. [0122]
  • Examples of useful dermatological compositions which can be used to deliver the compounds of formula I to the skin are known to the art; for example, see Jacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S. Pat. No. 4,992,478), Smith et al. (U.S. Pat. No. 4,559,157) and Wortzman (U.S. Pat. No. 4,820,508). [0123]
  • Useful dosages of the compounds of formula I can be determined by comparing their in vitro activity, and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art; for example, see U.S. Pat. No. 4,938,949. [0124]
  • The compound is conveniently administered in unit dosage form; for example, containing 5 to 1000 mg, conveniently 10 to 750 mg, most conveniently, 50 to 500 mg of active ingredient per unit dosage form. The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations; such as multiple inhalations from an insufflator or by application of a plurality of drops into the eye. [0125]
  • For internal infections, the compositions can be administered orally or parenterally at dose levels, calculated as the free base, of about 0.1 to 300 mg/kg, preferably 1.0 to 30 mg/kg of mammal body weight, and can be used in man in a unit dosage form, administered one to four times daily in the amount of 1 to 1000 mg per unit dose. [0126]
  • For parenteral administration or for administration as drops, as for eye infections, the compounds are presented in aqueous solution in a concentration of from about 0.1 to about 10%, more preferably about 0.1 to about 7%. The solution may contain other ingredients, such as emulsifiers, antioxidants or buffers. [0127]
  • Generally, the concentration of the compound(s) of formula I in a liquid composition, such as a lotion, will be from about 0.1-25 wt-%, preferably from about 0.5-10 wt-%. The concentration in a semi-solid or solid composition such as a gel or a powder will be about 0.1-5 wt-%, preferably about 0.5-2.5 wt-%. [0128]
  • The exact regimen for administration of the compounds and compositions disclosed herein will necessarily be dependent upon the needs of the individual subject being treated, the type of treatment and, of course, the judgment of the attending practitioner. [0129]
  • The antiviral activity of a compound of the invention can be determined using pharmacological models which are well known to the art, or using Test A described below. [0130]
  • The compounds of formula (I) and pharmaceutically acceptable salts thereof are useful as antiviral agents. Thus, they are useful to combat viral infections in animals, including man. The compounds are generally active against herpes viruses, and are particularly useful against the varicella zoster virus, the Epstein-Barr virus, the herpes simplex virus, the human herpes virus type 8 (HHV-8) and the cytomegalovirus (CMV). [0131]
  • 1 19 1 3717 DNA herpes simplex 1 atgttttgtg ccgcgggcgg cccgacttcc cccgggggga agtcggcggc tcgggcggcg 60 tctgggtttt ttgcccccca caacccccgg ggagccaccc agacggcacc gccgccttgc 120 cgccggcaga acttctacaa cccccacctc gctcagaccg gaacgcagcc aaaggccccc 180 gggccggctc agcgccatac gtactacagc gagtgcgacg aatttcgatt tatcgccccg 240 cgttcgctgg acgaggacgc ccccgcggag cagcgcaccg gggtccacga cggccgcctc 300 cggcgcgccc ctaaggtgta ctgcgggggg gacgagcgcg acgtcctccg cgtgggcccg 360 gagggcttct ggccgcgtcg cttgcgcctg tggggcggtg cggaccatgc ccccaagggg 420 ttcgacccca ccgtcaccgt cttccacgtg tacgacatcc tggagcacgt ggaacacgcg 480 tacagcatgc gcgccgccca gctccacgag cgatttatgg acgccatcac gcccgccggg 540 accgtcatca cgcttctggg tctgaccccc gaaggccatc gcgtcgccgt tcacgtctac 600 ggcacgcggc agtactttta catgaacaag gcggaggtgg atcggcacct gcagtgccgt 660 gccccgcgcg atctctgcga gcgcctggcg gcggccctgc gcgagtcgcc gggggcgtcg 720 ttccgcggca tctccgcgga ccacttcgag gcggaggtgg tggagcgcgc cgacgtgtac 780 tattacgaaa cgcgcccgac cctgtactac cgcgtcttcg tgcgaagcgg gcgcgcgctg 840 gcctacctgt gcgacaactt ttgccccgcg atcaggaagt acgagggggg cgtcgacgcc 900 accacccggt ttatcctgga caacccgggg tttgtcacct tcggctggta ccgcctcaag 960 cccggccgcg ggaacgcgcc ggcccaaccg cgccccccga cggcgttcgg aacctcgagc 1020 gacgtcgagt ttaactgcac ggcggacaac ctggccgtcg agggggccat gtgtgacctg 1080 ccggcctaca agctcatgtg cttcgatatc gaatgcaagg ccggggggga ggacgagctg 1140 gcctttccgg tcgcggaacg cccggaagac ctcgtcatcc agatctcctg tctgctctac 1200 gacctgtcca ccaccgccct cgagcacatc ctcctgtttt cgctcggatc ctgcgacctc 1260 cccgagtccc acctcagcga tctcgcctcc aggggcctgc cggcccccgt cgtcctggag 1320 tttgacagcg aattcgagat gctgctggcc ttcatgacct tcgtcaagca gtacggcccc 1380 gagttcgtga ccgggtacaa catcatcaac ttcgactggc ccttcgtcct gaccaagctg 1440 acggagatct acaaggtccc gctcgacggg tacgggcgca tgaacggccg gggtgtgttc 1500 cgcgtgtggg acatcggcca gagccacttt cagaagcgca gcaagatcaa ggtgaacggg 1560 atggtgaaca tcgacatgta cggcatcatc accgacaagg tcaaactctc cagctacaag 1620 ctgaacgccg tcgccgaggc cgtcttgaag gacaagaaga aggatctgag ctaccgcgac 1680 atccccgcct actacgcctc cgggcccgcg cagcgcgggg tgatcggcga gtattgtgtg 1740 caggactcgc tgctggtcgg gcagctgttc ttcaagtttc tgccgcacct ggagctttcc 1800 gccgtcgcgc gcctggcggg catcaacatc acccgcacca tctacgacgg ccagcagatc 1860 cgcgtcttca cgtgcctcct gcgccttgcg ggccagaagg gcttcatcct gccggacacc 1920 caggggcggt ttcggggcct cgacaaggag gcgcccaagc gcccggccgt gcctcggggg 1980 gaaggggagc ggccggggga cgggaacggg gacgaggata aggacgacga cgaggacgag 2040 gacggggacg agcgcgagga ggtcgcgcgc gagaccgggg gccggcacgt tgggtaccag 2100 ggggcccggg tcctcgaccc cacctccggg tttcacgtcg accccgtggt ggtgtttgac 2160 tttgccagcc tgtaccccag catcatccag gcccacaacc tgtgcttcag tacgctctcc 2220 ctgcggcccg aggccgtcgc gcacctggag gcggaccggg actacctgga gatcgaggtg 2280 gggggccgac ggctgttctt cgtgaaggcc cacgtacgcg agagcctgct gagcatcctg 2340 ctgcgcgact ggctggccat gcgaaagcag atccgctcgc ggatccccca gagcaccccc 2400 gaggaggccg tcctcctcga caagcaacag gccgccatca aggtggtgtg caactcggtg 2460 tacgggttca ccggggcgca gcacggtctt ctgccctgcc tgcacgtggc cgccaccgtg 2520 acgaccatcg gccgcgagat gctcctcgcg acgcgcgcgt acgtgcacgc gcgctgggcg 2580 gagttcgatc agctgctggc cgactttccg gaggcggccg gcatgcgcgc ccccggtccg 2640 tactccatgc gcatcatcta cggggacacg gactccattt tcgttttgtg ccgcggcctc 2700 acggccgcgg gcctggtggc catgggcgac aagatggcga gccacatctc gcgcgcgctg 2760 ttcctccccc cgatcaagct cgagtgcgaa aaaacgttca ccaagctgct gctcatcgcc 2820 aagaaaaagt acatcggcgt catctgcggg ggcaagatgc tcatcaaggg cgtggatctg 2880 gtgcgcaaaa acaactgcgc gtttatcaac cgcacctcca gggccctggt cgacctgctg 2940 ttttacgacg ataccgtatc cggagcggcc gccgcgttag ccgagcgccc cgcagaggag 3000 tggctggcgc gacccctgcc cgagggactg caggcgttcg gggccgtcct cgtagacgcc 3060 catcggcgca tcaccgaccc ggagagggac atccaggact ttgtcctcac cgccgaactg 3120 agcagacacc cgcgcgcgta caccaacaag cgcctggccc acctgacggt gtattacaag 3180 ctcatggccc gccgcgcgca ggtcccgtcc atcaaggacc ggatcccgta cgtgatcgtg 3240 gcccagaccc gcgaggtaga ggagacggtc gcgcggctgg ccgccctccg cgagctagac 3300 gccgccgccc caggggacga gcccgccccc ccagcggccc tgccctcccc ggccaagcgc 3360 ccccgggaga cgccgtcgca tgccgacccc ccgggaggcg cgtccaagcc ccgcaagctg 3420 ctggtgtccg agctggcgga ggatcccggg tacgccatcg cccggggcgt tccgctcaac 3480 acggactatt acttctcgca cctgctgggg gcggcctgcg tgacgttcaa ggccctgttt 3540 ggaaataacg ccaagatcac cgagagtctg ttaaagaggt ttattcccga gacgtggcac 3600 cccccggacg acgtggccgc gcggctcagg gccgcggggt tcgggccggc gggggccggc 3660 gctacggcgg aggaaactcg tcgaatgttg catagagcct ttgatactct agcatga 3717 2 1238 PRT herpes simplex 2 Met Phe Cys Ala Ala Gly Gly Pro Thr Ser Pro Gly Gly Lys Ser Ala 1 5 10 15 Ala Arg Ala Ala Ser Gly Phe Phe Ala Pro His Asn Pro Arg Gly Ala 20 25 30 Thr Gln Thr Ala Pro Pro Pro Cys Arg Arg Gln Asn Phe Tyr Asn Pro 35 40 45 His Leu Ala Gln Thr Gly Thr Gln Pro Lys Ala Pro Gly Pro Ala Gln 50 55 60 Arg His Thr Tyr Tyr Ser Glu Cys Asp Glu Phe Arg Phe Ile Ala Pro 65 70 75 80 Arg Ser Leu Asp Glu Asp Ala Pro Ala Glu Gln Arg Thr Gly Val His 85 90 95 Asp Gly Arg Leu Arg Arg Ala Pro Lys Val Tyr Cys Gly Gly Asp Glu 100 105 110 Arg Asp Val Leu Arg Val Gly Pro Glu Gly Phe Trp Pro Arg Arg Leu 115 120 125 Arg Leu Trp Gly Gly Ala Asp His Ala Pro Lys Gly Phe Asp Pro Thr 130 135 140 Val Thr Val Phe His Val Tyr Asp Ile Leu Glu His Val Glu His Ala 145 150 155 160 Tyr Ser Met Arg Ala Ala Gln Leu His Glu Arg Phe Met Asp Ala Ile 165 170 175 Thr Pro Ala Gly Thr Val Ile Thr Leu Leu Gly Leu Thr Pro Glu Gly 180 185 190 His Arg Val Ala Val His Val Tyr Gly Thr Arg Gln Tyr Phe Tyr Met 195 200 205 Asn Lys Ala Glu Val Asp Arg His Leu Gln Cys Arg Ala Pro Arg Asp 210 215 220 Leu Cys Glu Arg Leu Ala Ala Ala Leu Arg Glu Ser Pro Gly Ala Ser 225 230 235 240 Phe Arg Gly Ile Ser Ala Asp His Phe Glu Ala Glu Val Val Glu Arg 245 250 255 Ala Asp Val Tyr Tyr Tyr Glu Thr Arg Pro Thr Leu Tyr Tyr Arg Val 260 265 270 Phe Val Arg Ser Gly Arg Ala Leu Ala Tyr Leu Cys Asp Asn Phe Cys 275 280 285 Pro Ala Ile Arg Lys Tyr Glu Gly Gly Val Asp Ala Thr Thr Arg Phe 290 295 300 Ile Leu Asp Asn Pro Gly Phe Val Thr Phe Gly Trp Tyr Arg Leu Lys 305 310 315 320 Pro Gly Arg Gly Asn Ala Pro Ala Gln Pro Arg Pro Pro Thr Ala Phe 325 330 335 Gly Thr Ser Ser Asp Val Glu Phe Asn Cys Thr Ala Asp Asn Leu Ala 340 345 350 Val Glu Gly Ala Met Cys Asp Leu Pro Ala Tyr Lys Leu Met Cys Phe 355 360 365 Asp Ile Glu Cys Lys Ala Gly Gly Glu Asp Glu Leu Ala Phe Pro Val 370 375 380 Ala Glu Arg Pro Glu Asp Leu Val Ile Gln Ile Ser Cys Leu Leu Tyr 385 390 395 400 Asp Leu Ser Thr Thr Ala Leu Glu His Ile Leu Leu Phe Ser Leu Gly 405 410 415 Ser Cys Asp Leu Pro Glu Ser His Leu Ser Asp Leu Ala Ser Arg Gly 420 425 430 Leu Pro Ala Pro Val Val Leu Glu Phe Asp Ser Glu Phe Glu Met Leu 435 440 445 Leu Ala Phe Met Thr Phe Val Lys Gln Tyr Gly Pro Glu Phe Val Thr 450 455 460 Gly Tyr Asn Ile Ile Asn Phe Asp Trp Pro Phe Val Leu Thr Lys Leu 465 470 475 480 Thr Glu Ile Tyr Lys Val Pro Leu Asp Gly Tyr Gly Arg Met Asn Gly 485 490 495 Arg Gly Val Phe Arg Val Trp Asp Ile Gly Gln Ser His Phe Gln Lys 500 505 510 Arg Ser Lys Ile Lys Val Asn Gly Met Val Asn Ile Asp Met Tyr Gly 515 520 525 Ile Ile Thr Asp Lys Val Lys Leu Ser Ser Tyr Lys Leu Asn Ala Val 530 535 540 Ala Glu Ala Val Leu Lys Asp Lys Lys Lys Asp Leu Ser Tyr Arg Asp 545 550 555 560 Ile Pro Ala Tyr Tyr Ala Ser Gly Pro Ala Gln Arg Gly Val Ile Gly 565 570 575 Glu Tyr Cys Val Gln Asp Ser Leu Leu Val Gly Gln Leu Phe Phe Lys 580 585 590 Phe Leu Pro His Leu Glu Leu Ser Ala Val Ala Arg Leu Ala Gly Ile 595 600 605 Asn Ile Thr Arg Thr Ile Tyr Asp Gly Gln Gln Ile Arg Val Phe Thr 610 615 620 Cys Leu Leu Arg Leu Ala Gly Gln Lys Gly Phe Ile Leu Pro Asp Thr 625 630 635 640 Gln Gly Arg Phe Arg Gly Leu Asp Lys Glu Ala Pro Lys Arg Pro Ala 645 650 655 Val Pro Arg Gly Glu Gly Glu Arg Pro Gly Asp Gly Asn Gly Asp Glu 660 665 670 Asp Lys Asp Asp Asp Glu Asp Glu Asp Gly Asp Glu Arg Glu Glu Val 675 680 685 Ala Arg Glu Thr Gly Gly Arg His Val Gly Tyr Gln Gly Ala Arg Val 690 695 700 Leu Asp Pro Thr Ser Gly Phe His Val Asp Pro Val Val Val Phe Asp 705 710 715 720 Phe Ala Ser Leu Tyr Pro Ser Ile Ile Gln Ala His Asn Leu Cys Phe 725 730 735 Ser Thr Leu Ser Leu Arg Pro Glu Ala Val Ala His Leu Glu Ala Asp 740 745 750 Arg Asp Tyr Leu Glu Ile Glu Val Gly Gly Arg Arg Leu Phe Phe Val 755 760 765 Lys Ala His Val Arg Glu Ser Leu Leu Ser Ile Leu Leu Arg Asp Trp 770 775 780 Leu Ala Met Arg Lys Gln Ile Arg Ser Arg Ile Pro Gln Ser Thr Pro 785 790 795 800 Glu Glu Ala Val Leu Leu Asp Lys Gln Gln Ala Ala Ile Lys Val Val 805 810 815 Cys Asn Ser Val Tyr Gly Phe Thr Gly Ala Gln His Gly Leu Leu Pro 820 825 830 Cys Leu His Val Ala Ala Thr Val Thr Thr Ile Gly Arg Glu Met Leu 835 840 845 Leu Ala Thr Arg Ala Tyr Val His Ala Arg Trp Ala Glu Phe Asp Gln 850 855 860 Leu Leu Ala Asp Phe Pro Glu Ala Ala Gly Met Arg Ala Pro Gly Pro 865 870 875 880 Tyr Ser Met Arg Ile Ile Tyr Gly Asp Thr Asp Ser Ile Phe Val Leu 885 890 895 Cys Arg Gly Leu Thr Ala Ala Gly Leu Val Ala Met Gly Asp Lys Met 900 905 910 Ala Ser His Ile Ser Arg Ala Leu Phe Leu Pro Pro Ile Lys Leu Glu 915 920 925 Cys Glu Lys Thr Phe Thr Lys Leu Leu Leu Ile Ala Lys Lys Lys Tyr 930 935 940 Ile Gly Val Ile Cys Gly Gly Lys Met Leu Ile Lys Gly Val Asp Leu 945 950 955 960 Val Arg Lys Asn Asn Cys Ala Phe Ile Asn Arg Thr Ser Arg Ala Leu 965 970 975 Val Asp Leu Leu Phe Tyr Asp Asp Thr Val Ser Gly Ala Ala Ala Ala 980 985 990 Leu Ala Glu Arg Pro Ala Glu Glu Trp Leu Ala Arg Pro Leu Pro Glu 995 1000 1005 Gly Leu Gln Ala Phe Gly Ala Val Leu Val Asp Ala His Arg Arg 1010 1015 1020 Ile Thr Asp Pro Glu Arg Asp Ile Gln Asp Phe Val Leu Thr Ala 1025 1030 1035 Glu Leu Ser Arg His Pro Arg Ala Tyr Thr Asn Lys Arg Leu Ala 1040 1045 1050 His Leu Thr Val Tyr Tyr Lys Leu Met Ala Arg Arg Ala Gln Val 1055 1060 1065 Pro Ser Ile Lys Asp Arg Ile Pro Tyr Val Ile Val Ala Gln Thr 1070 1075 1080 Arg Glu Val Glu Glu Thr Val Ala Arg Leu Ala Ala Leu Arg Glu 1085 1090 1095 Leu Asp Ala Ala Ala Pro Gly Asp Glu Pro Ala Pro Pro Ala Ala 1100 1105 1110 Leu Pro Ser Pro Ala Lys Arg Pro Arg Glu Thr Pro Ser His Ala 1115 1120 1125 Asp Pro Pro Gly Gly Ala Ser Lys Pro Arg Lys Leu Leu Val Ser 1130 1135 1140 Glu Leu Ala Glu Asp Pro Gly Tyr Ala Ile Ala Arg Gly Val Pro 1145 1150 1155 Leu Asn Thr Asp Tyr Tyr Phe Ser His Leu Leu Gly Ala Ala Cys 1160 1165 1170 Val Thr Phe Lys Ala Leu Phe Gly Asn Asn Ala Lys Ile Thr Glu 1175 1180 1185 Ser Leu Leu Lys Arg Phe Ile Pro Glu Thr Trp His Pro Pro Asp 1190 1195 1200 Asp Val Ala Ala Arg Leu Arg Ala Ala Gly Phe Gly Pro Ala Gly 1205 1210 1215 Ala Gly Ala Thr Ala Glu Glu Thr Arg Arg Met Leu His Arg Ala 1220 1225 1230 Phe Asp Thr Leu Ala 1235 3 3723 DNA herpes simplex 3 atgttttgtg ccgcgggcgg cccggcttcc cccgggggga agtcggcggc tcgggcggcg 60 tctgggtttt ttgcccccca caacccccgg ggagccaccc agacggcacc gccgccttgc 120 cgccggcaga acttctacaa cccccacctc gctcagaccg gaacgcagcc aaaggccccc 180 gggccggctc agcgccatac gtactacagc gagtgcgacg aatttcgatt tatcgccccg 240 cgttcgctgg acgaggacgc ccccgcggag cagcgcaccg gggtccacga cggccgcctc 300 cggcgcgccc ctaaggtgta ctgcgggggg gacgagcgcg acgtcctccg cgtgggcccg 360 gagggcttct ggccgcgtcg cttgcgcctg tggggcggtg cggaccatgc ccccgagggg 420 ttcgacccca ccgtcaccgt cttccacgtg tacgacatcc tggagcacgt ggaacacgcg 480 tacagcatgc gcgccgccca gctccacgag cgatttatgg acgccatcac gcccgccggg 540 accgtcatca cgcttctggg tctgaccccc gaaggccatc gcgtcgccgt tcacgtctac 600 ggcacgcggc agtactttta catgaacaag gcggaggtgg atcggcacct gcagtgccgt 660 gccccgcgcg atctctgcga gcgcctggcg gcggccctgc gcgagtcgcc gggggcgtcg 720 ttccgcggca tctccgcgga ccacttcgag gcggaggtgg tggagcgcgc cgacgtgtac 780 tattacgaaa cgcgcccgac cctgtactac cgcgtcttcg tgcgaagcgg gcgcgcgctg 840 gcctacctgt gcgacaactt ttgccccgcg atcaggaagt acgagggggg cgtcgacgcc 900 accacccggt ttatcctgga caacccgggg tttgtcacct tcggctggta ccgcctcaag 960 cccggccgcg ggaacgcgcc ggcccaaccg cgccccccga cggcgttcgg aacctcgagc 1020 gacgtcgagt ttaactgcac ggcggacaac ctggccgtcg agggggccat gtgtgacctg 1080 ccggcctaca agctcatgtg cttcgatatc gaatgcaagg ccggggggga ggacgagctg 1140 gcctttccgg tcgcggaacg cccggaagac ctcgtcatcc agatctcctg tctgctctac 1200 gacctgtcca ccaccgccct cgagcacatc ctcctgtttt cgctcggatc ctgcgacctc 1260 cccgagtccc acctcagcga tctcgcctcc aggggcctgc cggcccccgt cgtcctggag 1320 tttgacagcg aattcgagat gctgctggcc ttcatgacct tcgtcaagca gtacggcccc 1380 gagttcgtga ccgggtacaa catcatcaac ttcgactggc ccttcgtcct gaccaagctg 1440 acggagatct acaaggtccc gctcgacggg tacgggcgca tgaacggccg gggtgtgttc 1500 cgcgtgtggg acatcggcca gagccacttt cagaagcgca gcaagatcaa ggtgaacggg 1560 atggtgaaca tcgacatgta cggcatcatc accgacaagg tcaaactctc cagctacaag 1620 ctgaacgccg tcgccgaggc cgtcttgaag gacaagaaga aggatctgag ctaccgcgac 1680 atccccgcct actacgcctc cgggcccgcg cagcgcgggg tgatcggcga gtattgtgtg 1740 caggactcgc tgctggtcgg gcagctgttc ttcaagtttc tgccgcacct ggagctttcc 1800 gccgtcgcgc gcctggcggg catcaacatc acccgcacca tctacgacgg ccagcagatc 1860 cgcgtcttca cgtgcctcct gcgccttgcg ggccagaagg gcttcatcct gccggacacc 1920 caggggcggt ttcggggcct cgacaaggag gcgcccaagc gcccggccgt gcctcggggg 1980 gaaggggagc ggccggggga cgggaacggg gacgaggata aggacgacga cgaggacggg 2040 gacgaggacg gggacgagcg cgaggaggtc gcgcgcgaga ccgggggccg gcacgttggg 2100 taccaggggg cccgggtcct cgaccccacc tccgggtttc acgtcgaccc cgtggtggtg 2160 tttgactttg ccagcctgta ccccagcatc atccaggccc acaacctgtg cttcagtacg 2220 ctctccctgc ggcccgaggc cgtcgcgcac ctggaggcgg accgggacta cctggagatc 2280 gaggtggggg gccgacggct gttcttcgtg aaggcccacg tacgcgagag cctgctgagc 2340 atcctgctgc gcgactggct ggccatgcga aagcagatcc gctcgcggat cccccagagc 2400 ccccccgagg aggccgtcct cctcgacaag caacaggccg ccatcaaggt ggtgtgcaac 2460 tcggtgtacg ggttcaccgg ggcgcagcac ggtcttctgc cctgcctgca cgtggccgcc 2520 accgtgacga ccatcggccg cgagatgctc ctcgcgacgc gcgcgtacgt gcacgcgcgc 2580 tgggcggagt tcgatcagct gctggccgac tttccggagg cggccggcat gcgcgccccc 2640 ggtccgtact ccatgcgcat catctacggg gacacggact ccattttcgt tttgtgccgc 2700 ggcctcacgg ccgcgggcct ggtggccatg ggcgacaaga tggcgagcca catctcgcgc 2760 gcgctgttcc tccccccgat caagctcgag tgcgaaaaaa cgttcaccaa gctgctgctc 2820 atcgccaaga aaaagtacat cggcgtcatc tgcgggggca agatgctcat caagggcgtg 2880 gatctggtgc gcaaaaacaa ctgcgcgttt atcaaccgca cctccagggc cctggtcgac 2940 ctgctgtttt acgacgatac cgtatccgga gcggccgccg cgttagccga gcgccccgca 3000 gaggagtggc tggcgcgacc cctgcccgag ggactgcagg cgttcggggc cgtcctcgta 3060 gacgcccatc ggcgcatcac cgacccggag agggacatcc aggactttgt cctcaccgcc 3120 gaactgagca gacacccgcg cgcgtacacc aacaagcgcc tggcccacct gacggtgtat 3180 tacaagctca tggcccgccg cgcgcaggtc ccgtccatca aggaccggat cccgtacgtg 3240 atcgtggccc agacccgcga ggtagaggag acggtcgcgc ggctggccgc cctccgcgag 3300 ctagacgccg ccgccccagg ggacgagccc gcccccccag cggccctgcc ctccccggcc 3360 aagcgccccc gggagacgcc gtcgcatgcc gaccccccgg gaggcgcgtc caagccccgc 3420 aagctgctgg tgtccgagct ggcggaggat cccgggtacg ccatcgcccg gggcgttccg 3480 ctcaacacgg actattactt ctcgcacctg ctgggggcgg cctgcgtgac gttcaaggcc 3540 ctgtttggaa ataacgccaa gatcaccgag agtctgttaa agaggtttat tcccgagacg 3600 tggcaccccc cggacgacgt ggccgcgcgg ctcagggccg cggggttcgg gccggcgggg 3660 gccggcgcta cggcggagga aactcgtcga atgttgcata gagcctttga tactctagca 3720 tga 3723 4 1240 PRT herpes simplex 4 Met Phe Cys Ala Ala Gly Gly Pro Ala Ser Pro Gly Gly Lys Ser Ala 1 5 10 15 Ala Arg Ala Ala Ser Gly Phe Phe Ala Pro His Asn Pro Arg Gly Ala 20 25 30 Thr Gln Thr Ala Pro Pro Pro Cys Arg Arg Gln Asn Phe Tyr Asn Pro 35 40 45 His Leu Ala Gln Thr Gly Thr Gln Pro Lys Ala Pro Gly Pro Ala Gln 50 55 60 Arg His Thr Tyr Tyr Ser Glu Cys Asp Glu Phe Arg Phe Ile Ala Pro 65 70 75 80 Arg Ser Leu Asp Glu Asp Ala Pro Ala Glu Gln Arg Thr Gly Val His 85 90 95 Asp Gly Arg Leu Arg Arg Ala Pro Lys Val Tyr Cys Gly Gly Asp Glu 100 105 110 Arg Asp Val Leu Arg Val Gly Pro Glu Gly Phe Trp Pro Arg Arg Leu 115 120 125 Arg Leu Trp Gly Gly Ala Asp His Ala Pro Glu Gly Phe Asp Pro Thr 130 135 140 Val Thr Val Phe His Val Tyr Asp Ile Leu Glu His Val Glu His Ala 145 150 155 160 Tyr Ser Met Arg Ala Ala Gln Leu His Glu Arg Phe Met Asp Ala Ile 165 170 175 Thr Pro Ala Gly Thr Val Ile Thr Leu Leu Gly Leu Thr Pro Glu Gly 180 185 190 His Arg Val Ala Val His Val Tyr Gly Thr Arg Gln Tyr Phe Tyr Met 195 200 205 Asn Lys Ala Glu Val Asp Arg His Leu Gln Cys Arg Ala Pro Arg Asp 210 215 220 Leu Cys Glu Arg Leu Ala Ala Ala Leu Arg Glu Ser Pro Gly Ala Ser 225 230 235 240 Phe Arg Gly Ile Ser Ala Asp His Phe Glu Ala Glu Val Val Glu Arg 245 250 255 Ala Asp Val Tyr Tyr Tyr Glu Thr Arg Pro Thr Leu Tyr Tyr Arg Val 260 265 270 Phe Val Arg Ser Gly Arg Ala Leu Ala Tyr Leu Cys Asp Asn Phe Cys 275 280 285 Pro Ala Ile Arg Lys Tyr Glu Gly Gly Val Asp Ala Thr Thr Arg Phe 290 295 300 Ile Leu Asp Asn Pro Gly Phe Val Thr Phe Gly Trp Tyr Arg Leu Lys 305 310 315 320 Pro Gly Arg Gly Asn Ala Pro Ala Gln Pro Arg Pro Pro Thr Ala Phe 325 330 335 Gly Thr Ser Ser Asp Val Glu Phe Asn Cys Thr Ala Asp Asn Leu Ala 340 345 350 Val Glu Gly Ala Met Cys Asp Leu Pro Ala Tyr Lys Leu Met Cys Phe 355 360 365 Asp Ile Glu Cys Lys Ala Gly Gly Glu Asp Glu Leu Ala Phe Pro Val 370 375 380 Ala Glu Arg Pro Glu Asp Leu Val Ile Gln Ile Ser Cys Leu Leu Tyr 385 390 395 400 Asp Leu Ser Thr Thr Ala Leu Glu His Ile Leu Leu Phe Ser Leu Gly 405 410 415 Ser Cys Asp Leu Pro Glu Ser His Leu Ser Asp Leu Ala Ser Arg Gly 420 425 430 Leu Pro Ala Pro Val Val Leu Glu Phe Asp Ser Glu Phe Glu Met Leu 435 440 445 Leu Ala Phe Met Thr Phe Val Lys Gln Tyr Gly Pro Glu Phe Val Thr 450 455 460 Gly Tyr Asn Ile Ile Asn Phe Asp Trp Pro Phe Val Leu Thr Lys Leu 465 470 475 480 Thr Glu Ile Tyr Lys Val Pro Leu Asp Gly Tyr Gly Arg Met Asn Gly 485 490 495 Arg Gly Val Phe Arg Val Trp Asp Ile Gly Gln Ser His Phe Gln Lys 500 505 510 Arg Ser Lys Ile Lys Val Asn Gly Met Val Asn Ile Asp Met Tyr Gly 515 520 525 Ile Ile Thr Asp Lys Val Lys Leu Ser Ser Tyr Lys Leu Asn Ala Val 530 535 540 Ala Glu Ala Val Leu Lys Asp Lys Lys Lys Asp Leu Ser Tyr Arg Asp 545 550 555 560 Ile Pro Ala Tyr Tyr Ala Ser Gly Pro Ala Gln Arg Gly Val Ile Gly 565 570 575 Glu Tyr Cys Val Gln Asp Ser Leu Leu Val Gly Gln Leu Phe Phe Lys 580 585 590 Phe Leu Pro His Leu Glu Leu Ser Ala Val Ala Arg Leu Ala Gly Ile 595 600 605 Asn Ile Thr Arg Thr Ile Tyr Asp Gly Gln Gln Ile Arg Val Phe Thr 610 615 620 Cys Leu Leu Arg Leu Ala Gly Gln Lys Gly Phe Ile Leu Pro Asp Thr 625 630 635 640 Gln Gly Arg Phe Arg Gly Leu Asp Lys Glu Ala Pro Lys Arg Pro Ala 645 650 655 Val Pro Arg Gly Glu Gly Glu Arg Pro Gly Asp Gly Asn Gly Asp Glu 660 665 670 Asp Lys Asp Asp Asp Glu Asp Gly Asp Glu Asp Gly Asp Glu Arg Glu 675 680 685 Glu Val Ala Arg Glu Thr Gly Gly Arg His Val Gly Tyr Gln Gly Ala 690 695 700 Arg Val Leu Asp Pro Thr Ser Gly Phe His Val Asp Pro Val Val Val 705 710 715 720 Phe Asp Phe Ala Ser Leu Tyr Pro Ser Ile Ile Gln Ala His Asn Leu 725 730 735 Cys Phe Ser Thr Leu Ser Leu Arg Pro Glu Ala Val Ala His Leu Glu 740 745 750 Ala Asp Arg Asp Tyr Leu Glu Ile Glu Val Gly Gly Arg Arg Leu Phe 755 760 765 Phe Val Lys Ala His Val Arg Glu Ser Leu Leu Ser Ile Leu Leu Arg 770 775 780 Asp Trp Leu Ala Met Arg Lys Gln Ile Arg Ser Arg Ile Pro Gln Ser 785 790 795 800 Pro Pro Glu Glu Ala Val Leu Leu Asp Lys Gln Gln Ala Ala Ile Lys 805 810 815 Val Val Cys Asn Ser Val Tyr Gly Phe Thr Gly Ala Gln His Gly Leu 820 825 830 Leu Pro Cys Leu His Val Ala Ala Thr Val Thr Thr Ile Gly Arg Glu 835 840 845 Met Leu Leu Ala Thr Arg Ala Tyr Val His Ala Arg Trp Ala Glu Phe 850 855 860 Asp Gln Leu Leu Ala Asp Phe Pro Glu Ala Ala Gly Met Arg Ala Pro 865 870 875 880 Gly Pro Tyr Ser Met Arg Ile Ile Tyr Gly Asp Thr Asp Ser Ile Phe 885 890 895 Val Leu Cys Arg Gly Leu Thr Ala Ala Gly Leu Val Ala Met Gly Asp 900 905 910 Lys Met Ala Ser His Ile Ser Arg Ala Leu Phe Leu Pro Pro Ile Lys 915 920 925 Leu Glu Cys Glu Lys Thr Phe Thr Lys Leu Leu Leu Ile Ala Lys Lys 930 935 940 Lys Tyr Ile Gly Val Ile Cys Gly Gly Lys Met Leu Ile Lys Gly Val 945 950 955 960 Asp Leu Val Arg Lys Asn Asn Cys Ala Phe Ile Asn Arg Thr Ser Arg 965 970 975 Ala Leu Val Asp Leu Leu Phe Tyr Asp Asp Thr Val Ser Gly Ala Ala 980 985 990 Ala Ala Leu Ala Glu Arg Pro Ala Glu Glu Trp Leu Ala Arg Pro Leu 995 1000 1005 Pro Glu Gly Leu Gln Ala Phe Gly Ala Val Leu Val Asp Ala His 1010 1015 1020 Arg Arg Ile Thr Asp Pro Glu Arg Asp Ile Gln Asp Phe Val Leu 1025 1030 1035 Thr Ala Glu Leu Ser Arg His Pro Arg Ala Tyr Thr Asn Lys Arg 1040 1045 1050 Leu Ala His Leu Thr Val Tyr Tyr Lys Leu Met Ala Arg Arg Ala 1055 1060 1065 Gln Val Pro Ser Ile Lys Asp Arg Ile Pro Tyr Val Ile Val Ala 1070 1075 1080 Gln Thr Arg Glu Val Glu Glu Thr Val Ala Arg Leu Ala Ala Leu 1085 1090 1095 Arg Glu Leu Asp Ala Ala Ala Pro Gly Asp Glu Pro Ala Pro Pro 1100 1105 1110 Ala Ala Leu Pro Ser Pro Ala Lys Arg Pro Arg Glu Thr Pro Ser 1115 1120 1125 His Ala Asp Pro Pro Gly Gly Ala Ser Lys Pro Arg Lys Leu Leu 1130 1135 1140 Val Ser Glu Leu Ala Glu Asp Pro Gly Tyr Ala Ile Ala Arg Gly 1145 1150 1155 Val Pro Leu Asn Thr Asp Tyr Tyr Phe Ser His Leu Leu Gly Ala 1160 1165 1170 Ala Cys Val Thr Phe Lys Ala Leu Phe Gly Asn Asn Ala Lys Ile 1175 1180 1185 Thr Glu Ser Leu Leu Lys Arg Phe Ile Pro Glu Thr Trp His Pro 1190 1195 1200 Pro Asp Asp Val Ala Ala Arg Leu Arg Ala Ala Gly Phe Gly Pro 1205 1210 1215 Ala Gly Ala Gly Ala Thr Ala Glu Glu Thr Arg Arg Met Leu His 1220 1225 1230 Arg Ala Phe Asp Thr Leu Ala 1235 1240 5 3708 DNA herpes simplex 5 atgttttccg gtggcggcgg cccgctgtcc cccggaggaa agtcggcggc cagggcggcg 60 tccgggtttt ttgcgcccgc cggccctcgc ggagccggcc ggggaccccc gccttgtttg 120 aggcaaaact tttacaaccc ctacctcgcc ccagtcggga cgcaacagaa gccgaccggg 180 ccaacccagc gccatacgta ctatagcgaa tgcgatgaat ttcgattcat cgccccgcgg 240 gtgctggacg aggatgcccc cccggagaag cgcgccgggg tgcacgacgg tcacctcaag 300 cgcgccccca aggtgtactg cgggggggac gagcgcgacg tcctccgcgt cgggtcgggc 360 ggcttctggc cgcggcgctc gcgcctgtgg ggcggcgtgg accacgcccc ggcggggttc 420 aaccccaccg tcaccgtctt tcacgtgtac gacatcctgg agaacgtgga gcacgcgtac 480 ggcatgcgcg cggcccagtt ccacgcgcgg tttatggacg ccatcacacc gacggggacc 540 gtcatcacgc tcctgggcct gactccggaa ggccaccggg tggccgttca cgtttacggc 600 acgcggcagt acttttacat gaacaaggag gaggttgaca ggcacctaca atgccgcgcc 660 ccacgagatc tctgcgagcg catggccgcg gccctgcgcg agtccccggg cgcgtcgttc 720 cgcggcatct ccgcggacca cttcgaggcg gaggtggtgg agcgcaccga cgtgtactac 780 tacgagacgc gccccgctct gttttaccgc gtctacgtcc gaagcgggcg cgtgctgtcg 840 tacctgtgcg acaacttctg cccggccatc aagaagtacg agggtggggt cgacgccacc 900 acccggttca tcctggacaa ccccgggttc gtcaccttcg gctggtaccg tctcaaaccg 960 ggccggaaca acacgctagc ccagccgcgg gccccgatgg ccttcgggac atccagcgac 1020 gtcgagttta actgtacggc ggacaacctg gccatcgagg ggggcatgag cgacctaccg 1080 gcatacaagc tcatgtgctt cgatatcgaa tgcaaggcgg ggggggagga cgagctggcc 1140 tttccggtgg ccgggcaccc ggaggacctg gttattcaga tatcctgtct gctctacgac 1200 ctgtccacca ccgccctgga gcacgtcctc ctgttttcgc tcggttcctg cgacctcccc 1260 gaatcccacc tgaacgagct ggcggccagg ggcctgccca cgcccgtggt tctggaattc 1320 gacagcgaat tcgagatgct gttggccttc atgacccttg tgaaacagta cggccccgag 1380 ttcgtgaccg ggtacaacat catcaacttc gactggccct tcttgctggc caagttgacg 1440 gacatttaca aggtccccct ggacgggtac ggccgcatga acggccgggg cgtgtttcgc 1500 gtgtgggaca taggccagag ccacttccag aagcgcagca agataaaggt gaacggcatg 1560 gtgaacatcg acatgtacgg gatcataacc gacaagatca agctctcgag ctacaagctc 1620 aacgccgtgg ccgaagccgt cctgaaggac aagaagaagg acctgagcta tcgcgacatc 1680 cccgcctact acgccgccgg gcccgcgcaa cgcggggtga tcggcgagta ctgcatacag 1740 gattccctgc tggtgggcca gctgtttttt aagtttttgc cccatctgga gctctcggcc 1800 gtcgcgcgct tggcgggtat taacatcacc cgcaccatct acgacggcca gcagatccgc 1860 gtctttacgt gcctgctgcg cctggccgac cagaagggct ttattctgcc ggacacccag 1920 gggcgattta ggggcgccgg gggggaggcg cccaagcgtc cggccgcagc ccgggaggac 1980 gaggagcggc cagaggagga gggggaggac gaggacgaac gcgaggaggg cgggggcgag 2040 cgggagccgg agggcgcgcg ggagaccgcc ggccggcacg tggggtacca gggggccagg 2100 gtccttgacc ccacttccgg gtttcacgtg aaccccgtgg tggtgttcga ctttgccagc 2160 ctgtacccca gcatcatcca ggcccacaac ctgtgcttca gcacgctctc cctgagggcc 2220 gacgcagtgg cgcacctgga ggcgggcaag gactacctgg agatcgaggt gggggggcga 2280 cggctgttct tcgtcaaggc tcacgtgcga gagagcctcc tcagcatcct cctgcgggac 2340 tggctcgcca tgcgaaagca gatccgctcg cggattcccc agagcagccc cgaggaggcc 2400 gtgctcctgg acaagcagca ggccgccatc aaggtcgtgt gtaactcggt gtacgggttc 2460 acgggagcgc agcacggact cctgccgtgc ctgcacgttg ccgcgacggt gacgaccatc 2520 ggccgcgaga tgctgctcgc gacccgcgag tacgtccacg cgcgctgggc ggccttcgaa 2580 cagctcctgg ccgatttccc ggaggcggcc gacatgcgcg cccccgggcc ctattccatg 2640 cgcatcatct acggggacac ggactccata tttgtgctgt gccgcggcct cacggccgcc 2700 gggctgacgg ccatgggcga caagatggcg agccacatct cgcgcgcgct gtttctgccc 2760 cccatcaaac tcgagtgcga aaagacgttc accaagctgc tgctgatcgc caagaaaaag 2820 tacatcggcg tcatctacgg gggtaagatg ctcatcaagg gcgtggatct ggtgcgcaaa 2880 aacaactgcg cgtttatcaa ccgcacctcc agggccctgg tcgacctgct gttttacgac 2940 gataccgtat ccggagcggc cgccgcgtta gccgagcgcc ccgcagagga gtggctggcg 3000 cgacccctgc ccgagggact gcaggcgttc ggggccgtcc tcgtagacgc ccatcggcgc 3060 atcaccgacc cggagaggga catccaggac tttgtcctca ccgccgaact gagcagacac 3120 ccgcgcgcgt acaccaacaa gcgcctggcc cacctgacgg tgtattacaa gctcatggcc 3180 cgccgcgcgc aggtcccgtc catcaaggac cggatcccgt acgtgatcgt ggcccagacc 3240 cgcgaggtag aggagacggt cgcgcggctg gccgccctcc gcgagctaga cgccgccgcc 3300 ccaggggacg agcccgcccc ccccgcggcc ctgccctccc cggccaagcg cccccgggag 3360 acgccgtcgc atgccgaccc cccgggaggc gcgtccaagc cccgcaagct gctggtgtcc 3420 gagctggccg aggatcccgc atacgccatt gcccacggcg tcgccctgaa cacggactat 3480 tacttctccc acctgttggg ggcggcgtgc gtgacattca aggccctgtt tgggaataac 3540 gccaagatca ccgagagtct gttaaaaagg tttattcccg aagtgtggca ccccccggac 3600 gacgtggccg cgcggctccg ggccgcaggg ttcggggcgg tgggtgccgg cgctacggcg 3660 gaggaaactc gtcgaatgtt gcatagagcc tttgatactc tagcatga 3708 6 1235 PRT herpes simplex 6 Met Phe Ser Gly Gly Gly Gly Pro Leu Ser Pro Gly Gly Lys Ser Ala 1 5 10 15 Ala Arg Ala Ala Ser Gly Phe Phe Ala Pro Ala Gly Pro Arg Gly Ala 20 25 30 Gly Arg Gly Pro Pro Pro Cys Leu Arg Gln Asn Phe Tyr Asn Pro Tyr 35 40 45 Leu Ala Pro Val Gly Thr Gln Gln Lys Pro Thr Gly Pro Thr Gln Arg 50 55 60 His Thr Tyr Tyr Ser Glu Cys Asp Glu Phe Arg Phe Ile Ala Pro Arg 65 70 75 80 Val Leu Asp Glu Asp Ala Pro Pro Glu Lys Arg Ala Gly Val His Asp 85 90 95 Gly His Leu Lys Arg Ala Pro Lys Val Tyr Cys Gly Gly Asp Glu Arg 100 105 110 Asp Val Leu Arg Val Gly Ser Gly Gly Phe Trp Pro Arg Arg Ser Arg 115 120 125 Leu Trp Gly Gly Val Asp His Ala Pro Ala Gly Phe Asn Pro Thr Val 130 135 140 Thr Val Phe His Val Tyr Asp Ile Leu Glu Asn Val Glu His Ala Tyr 145 150 155 160 Gly Met Arg Ala Ala Gln Phe His Ala Arg Phe Met Asp Ala Ile Thr 165 170 175 Pro Thr Gly Thr Val Ile Thr Leu Leu Gly Leu Thr Pro Glu Gly His 180 185 190 Arg Val Ala Val His Val Tyr Gly Thr Arg Gln Tyr Phe Tyr Met Asn 195 200 205 Lys Glu Glu Val Asp Arg His Leu Gln Cys Arg Ala Pro Arg Asp Leu 210 215 220 Cys Glu Arg Met Ala Ala Ala Leu Arg Glu Ser Pro Gly Ala Ser Phe 225 230 235 240 Arg Gly Ile Ser Ala Asp His Phe Glu Ala Glu Val Val Glu Arg Thr 245 250 255 Asp Val Tyr Tyr Tyr Glu Thr Arg Pro Ala Leu Phe Tyr Arg Val Tyr 260 265 270 Val Arg Ser Gly Arg Val Leu Ser Tyr Leu Cys Asp Asn Phe Cys Pro 275 280 285 Ala Ile Lys Lys Tyr Glu Gly Gly Val Asp Ala Thr Thr Arg Phe Ile 290 295 300 Leu Asp Asn Pro Gly Phe Val Thr Phe Gly Trp Tyr Arg Leu Lys Pro 305 310 315 320 Gly Arg Asn Asn Thr Leu Ala Gln Pro Arg Ala Pro Met Ala Phe Gly 325 330 335 Thr Ser Ser Asp Val Glu Phe Asn Cys Thr Ala Asp Asn Leu Ala Ile 340 345 350 Glu Gly Gly Met Ser Asp Leu Pro Ala Tyr Lys Leu Met Cys Phe Asp 355 360 365 Ile Glu Cys Lys Ala Gly Gly Glu Asp Glu Leu Ala Phe Pro Val Ala 370 375 380 Gly His Pro Glu Asp Leu Val Ile Gln Ile Ser Cys Leu Leu Tyr Asp 385 390 395 400 Leu Ser Thr Thr Ala Leu Glu His Val Leu Leu Phe Ser Leu Gly Ser 405 410 415 Cys Asp Leu Pro Glu Ser His Leu Asn Glu Leu Ala Ala Arg Gly Leu 420 425 430 Pro Thr Pro Val Val Leu Glu Phe Asp Ser Glu Phe Glu Met Leu Leu 435 440 445 Ala Phe Met Thr Leu Val Lys Gln Tyr Gly Pro Glu Phe Val Thr Gly 450 455 460 Tyr Asn Ile Ile Asn Phe Asp Trp Pro Phe Leu Leu Ala Lys Leu Thr 465 470 475 480 Asp Ile Tyr Lys Val Pro Leu Asp Gly Tyr Gly Arg Met Asn Gly Arg 485 490 495 Gly Val Phe Arg Val Trp Asp Ile Gly Gln Ser His Phe Gln Lys Arg 500 505 510 Ser Lys Ile Lys Val Asn Gly Met Val Asn Ile Asp Met Tyr Gly Ile 515 520 525 Ile Thr Asp Lys Ile Lys Leu Ser Ser Tyr Lys Leu Asn Ala Val Ala 530 535 540 Glu Ala Val Leu Lys Asp Lys Lys Lys Asp Leu Ser Tyr Arg Asp Ile 545 550 555 560 Pro Ala Tyr Tyr Ala Ala Gly Pro Ala Gln Arg Gly Val Ile Gly Glu 565 570 575 Tyr Cys Ile Gln Asp Ser Leu Leu Val Gly Gln Leu Phe Phe Lys Phe 580 585 590 Leu Pro His Leu Glu Leu Ser Ala Val Ala Arg Leu Ala Gly Ile Asn 595 600 605 Ile Thr Arg Thr Ile Tyr Asp Gly Gln Gln Ile Arg Val Phe Thr Cys 610 615 620 Leu Leu Arg Leu Ala Asp Gln Lys Gly Phe Ile Leu Pro Asp Thr Gln 625 630 635 640 Gly Arg Phe Arg Gly Ala Gly Gly Glu Ala Pro Lys Arg Pro Ala Ala 645 650 655 Ala Arg Glu Asp Glu Glu Arg Pro Glu Glu Glu Gly Glu Asp Glu Asp 660 665 670 Glu Arg Glu Glu Gly Gly Gly Glu Arg Glu Pro Glu Gly Ala Arg Glu 675 680 685 Thr Ala Gly Arg His Val Gly Tyr Gln Gly Ala Arg Val Leu Asp Pro 690 695 700 Thr Ser Gly Phe His Val Asn Pro Val Val Val Phe Asp Phe Ala Ser 705 710 715 720 Leu Tyr Pro Ser Ile Ile Gln Ala His Asn Leu Cys Phe Ser Thr Leu 725 730 735 Ser Leu Arg Ala Asp Ala Val Ala His Leu Glu Ala Gly Lys Asp Tyr 740 745 750 Leu Glu Ile Glu Val Gly Gly Arg Arg Leu Phe Phe Val Lys Ala His 755 760 765 Val Arg Glu Ser Leu Leu Ser Ile Leu Leu Arg Asp Trp Leu Ala Met 770 775 780 Arg Lys Gln Ile Arg Ser Arg Ile Pro Gln Ser Ser Pro Glu Glu Ala 785 790 795 800 Val Leu Leu Asp Lys Gln Gln Ala Ala Ile Lys Val Val Cys Asn Ser 805 810 815 Val Tyr Gly Phe Thr Gly Ala Gln His Gly Leu Leu Pro Cys Leu His 820 825 830 Val Ala Ala Thr Val Thr Thr Ile Gly Arg Glu Met Leu Leu Ala Thr 835 840 845 Arg Glu Tyr Val His Ala Arg Trp Ala Ala Phe Glu Gln Leu Leu Ala 850 855 860 Asp Phe Pro Glu Ala Ala Asp Met Arg Ala Pro Gly Pro Tyr Ser Met 865 870 875 880 Arg Ile Ile Tyr Gly Asp Thr Asp Ser Ile Phe Val Leu Cys Arg Gly 885 890 895 Leu Thr Ala Ala Gly Leu Thr Ala Met Gly Asp Lys Met Ala Ser His 900 905 910 Ile Ser Arg Ala Leu Phe Leu Pro Pro Ile Lys Leu Glu Cys Glu Lys 915 920 925 Thr Phe Thr Lys Leu Leu Leu Ile Ala Lys Lys Lys Tyr Ile Gly Val 930 935 940 Ile Tyr Gly Gly Lys Met Leu Ile Lys Gly Val Asp Leu Val Arg Lys 945 950 955 960 Asn Asn Cys Ala Phe Ile Asn Arg Thr Ser Arg Ala Leu Val Asp Leu 965 970 975 Leu Phe Tyr Asp Asp Thr Val Ser Gly Ala Ala Ala Ala Leu Ala Glu 980 985 990 Arg Pro Ala Glu Glu Trp Leu Ala Arg Pro Leu Pro Glu Gly Leu Gln 995 1000 1005 Ala Phe Gly Ala Val Leu Val Asp Ala His Arg Arg Ile Thr Asp 1010 1015 1020 Pro Glu Arg Asp Ile Gln Asp Phe Val Leu Thr Ala Glu Leu Ser 1025 1030 1035 Arg His Pro Arg Ala Tyr Thr Asn Lys Arg Leu Ala His Leu Thr 1040 1045 1050 Val Tyr Tyr Lys Leu Met Ala Arg Arg Ala Gln Val Pro Ser Ile 1055 1060 1065 Lys Asp Arg Ile Pro Tyr Val Ile Val Ala Gln Thr Arg Glu Val 1070 1075 1080 Glu Glu Thr Val Ala Arg Leu Ala Ala Leu Arg Glu Leu Asp Ala 1085 1090 1095 Ala Ala Pro Gly Asp Glu Pro Ala Pro Pro Ala Ala Leu Pro Ser 1100 1105 1110 Pro Ala Lys Arg Pro Arg Glu Thr Pro Ser His Ala Asp Pro Pro 1115 1120 1125 Gly Gly Ala Ser Lys Pro Arg Lys Leu Leu Val Ser Glu Leu Ala 1130 1135 1140 Glu Asp Pro Ala Tyr Ala Ile Ala His Gly Val Ala Leu Asn Thr 1145 1150 1155 Asp Tyr Tyr Phe Ser His Leu Leu Gly Ala Ala Cys Val Thr Phe 1160 1165 1170 Lys Ala Leu Phe Gly Asn Asn Ala Lys Ile Thr Glu Ser Leu Leu 1175 1180 1185 Lys Arg Phe Ile Pro Glu Val Trp His Pro Pro Asp Asp Val Ala 1190 1195 1200 Ala Arg Leu Arg Ala Ala Gly Phe Gly Ala Val Gly Ala Gly Ala 1205 1210 1215 Thr Ala Glu Glu Thr Arg Arg Met Leu His Arg Ala Phe Asp Thr 1220 1225 1230 Leu Ala 1235 7 3708 DNA herpes simplex 7 atgttttccg gtggcggcgg cccgctgtcc cccggaggaa agtcggcggc cagggcggcg 60 tccgggtttt ttgcgcccgc cggccctcgc ggagccggcc ggggaccccc gccttgcttg 120 aggcaaaact tttacaaccc ctacctcgcc ccagtcggga cgcaacagaa gccgaccggg 180 ccaacccagc gccatacgta ctatagcgaa tgcgatgaat ttcgattcat cgccccgcgg 240 gtgctggacg aggatgcccc cccggagaag cgcgccgggg tgcacgacgg tcacctcaag 300 cgcgccccca aggtgtactg cgggggggac gagcgcgacg tcctccgcgt cgggtcgggc 360 ggcttctggc cgcggcgctc gcgcctgtgg ggcggcgtgg accacgcccc ggcggggttc 420 aaccccaccg tcaccgtctt tcacgtgtac gacatcctgg agaacgtgga gcacgcgtac 480 ggcatgcgcg cggcccagtt ccacgcgcgg tttatggacg ccatcacacc gacggggacc 540 gtcatcacgc tcctgggcct gactccggaa ggccaccggg tggccgttca cgtttacggc 600 acgcggcagt acttttacat gaacaaggag gaggtcgaca ggcacctaca atgccgcgcc 660 ccacgagatc tctgcgagcg catggccgcg gccctgcgcg agtccccggg cgcgtcgttc 720 cgcggcattt ccgcggacca cttcgaggcg gaggtggtgg agcgcaccga cgtgtactac 780 tacgagacgc gccccgctct gttttaccgc gtctacgtcc gaagcgggcg cgtgctgtcg 840 tacctgtgcg acaacttctg cccggccatc aagaagtacg agggtggggt cgacgccacc 900 acccggttca tcctggacaa ccccgggttc gtcaccttcg gctggtaccg tctcaaaccg 960 ggccggaaca acacgctagc ccagccgcgg gccccgatgg ccttcgggac atccagcgac 1020 gtcgagttta actgtacggc ggacaacctg gccatcgagg ggggcatgag cgacctaccg 1080 gcatacaagc tcatgtgctt cgatatcgaa tgcaaggcgg ggggggagga cgagctggcc 1140 tttccggtgg ccgggcaccc ggaggacctg gtcatccaga tatcctgtct gctctacgac 1200 ctgtccacca ccgccctgga gcacgtcctc ctgttttcgc tcggttcctg cgacctcccc 1260 gaatcccacc tgaacgagct ggcggccagg ggcctgccca cgcccgtggt tctggaattc 1320 gacagcgaat tcgagatgct gttggccttc atgacccttg tgaaacagta cggccccgag 1380 ttcgtgaccg ggtacaacat catcaacttc gactggccct tcttgctggc caagctgacg 1440 gacatttaca aggtccccct ggacgggtac ggccgcatga acggccgggg cgtgtttcgc 1500 gtgtgggaca taggccagag ccacttccag aagcgcagca agataaaggt gaacggcatg 1560 gtgaacatcg acatgtacgg gattataacc gacaagatca agctctcgag ctacaagctc 1620 aacgccgtgg ccgaagccgt cctgaaggac aagaagaagg acctgagcta tcgcgacatc 1680 cccgcctact acgccgccgg gcccgcgcaa cgcggggtga tcggcgagta ctgcatacag 1740 gattccctgc tggtgggcca gctgtttttt aagtttttgc cccatctgga gctctcggcc 1800 gtcgcgcgct tggcgggtat taacatcacc cgcaccatct acgacggcca gcagatccgc 1860 gtctttacgt gcctgctgcg cctggccgac cagaagggct ttattctgcc ggacacccag 1920 gggcgattta ggggcggcgg gggggaggcg cccaagcgtc cggccgcagc ccgggaggac 1980 gaggagcggc cagaggagga gggggaggac gaggacgaac gcgaggaggg cgggggcgag 2040 cgggagccgg agggcgcgcg ggagaccgcc ggccggcacg tggggtacca gggggccagg 2100 gtccttgacc ccacttccgg gtttcatgtg aaccccgtgg tggtgttcga ctttgccagc 2160 ctgtacccca gcatcatcca ggcccacaac ctgtgcttca gcacgctctc cctgagggcc 2220 gacgcagtgg cgcacctgga ggcgggcaag gactacctgg agatcgaggt gggggggcga 2280 cggctgttct tcgtcaaggc tcacgtgcga gagagcctcc tcagcatcct cctgcgggac 2340 tggctcgcca tgcgaaagca gatccgctcg cggattcccc agagcagccc cgaggaggcc 2400 gtgctcctgg acaagcagca ggccgccatc aaggtcgtgt gtaactcggt ttacgggttc 2460 acgggagcgc agcacggact cctgccgtgc ctgcacgttg ccgcgacggt gacgaccatc 2520 ggccgcgaga tgctgctcgc gacccgcgag tacgtccacg cgcgctgggc ggccttcgaa 2580 cagctcctgg ccgatttccc ggaggcggcc gacatgcgcg cccccgggcc ctattccatg 2640 cgcatcatct acggggacac ggactccatc tttgtgctgt gccgcggcct cacggccgcc 2700 gggctgacgg ccgtgggcga caagatggcg agccacatct cgcgcgcgct gtttctgtcc 2760 cccatcaaac tcgagtgcga aaagacgttc accaagctgc tgctgatcgc caagaaaaag 2820 tacatcggcg tcatctacgg gggtaagatg ctcatcaagg gcgtggatct ggtgcgcaaa 2880 aacaactgcg cgtttatcaa ccgcacctcc agggccctgg tcgacctgct gttttacgac 2940 gataccgtat ccggagcggc cgccgcgtta gccgagcgcc ccgcagagga gtggctggcg 3000 cgacccctgc ccgagggact gcaggcgttc ggggccgtcc tcgtagacgc ccatcggcgc 3060 atcaccgacc cggagaggga catccaggac tttgtcctca ccgccgaact gagcagacac 3120 ccgcgcgcgt acaccaacaa gcgcctggcc cacctgacgg tgtattacaa gctcatggcc 3180 cgccgcgcgc aggtcccgtc catcaaggac cggatcccgt acgtgatcgt ggcccagacc 3240 cgcgaggtag aggagacggt cgcgcggctg gccgccctcc gcgagctcga cgccgccgcc 3300 ccaggggacg agcccgcccc ccccgcggcc ctgccctccc cggccaagcg cccccgggag 3360 acgccgttgc atgccgaccc cccgggaggc gcgtccaagc cccgcaagct gctggtgtcc 3420 gagctggccg aggatcccgc atacgccatt gcccacggcg tcgccctgaa cacggactat 3480 tacttctccc acctgttggg ggcggcgtgc gtgacattca aggccctgtt tgggaataac 3540 gccaagatca ccgagagtct gttaaaaagg tttattcccg aagtgtggca ccccccggac 3600 gacgtggccg cgcggctccg ggccgcaggg ttcggggcgg tgggtgccgg cgctacggcg 3660 gaggaaactc gtcgaatgtt gcatagagcc tttgatactc tagcatga 3708 8 1235 PRT herpes simplex 8 Met Phe Ser Gly Gly Gly Gly Pro Leu Ser Pro Gly Gly Lys Ser Ala 1 5 10 15 Ala Arg Ala Ala Ser Gly Phe Phe Ala Pro Ala Gly Pro Arg Gly Ala 20 25 30 Gly Arg Gly Pro Pro Pro Cys Leu Arg Gln Asn Phe Tyr Asn Pro Tyr 35 40 45 Leu Ala Pro Val Gly Thr Gln Gln Lys Pro Thr Gly Pro Thr Gln Arg 50 55 60 His Thr Tyr Tyr Ser Glu Cys Asp Glu Phe Arg Phe Ile Ala Pro Arg 65 70 75 80 Val Leu Asp Glu Asp Ala Pro Pro Glu Lys Arg Ala Gly Val His Asp 85 90 95 Gly His Leu Lys Arg Ala Pro Lys Val Tyr Cys Gly Gly Asp Glu Arg 100 105 110 Asp Val Leu Arg Val Gly Ser Gly Gly Phe Trp Pro Arg Arg Ser Arg 115 120 125 Leu Trp Gly Gly Val Asp His Ala Pro Ala Gly Phe Asn Pro Thr Val 130 135 140 Thr Val Phe His Val Tyr Asp Ile Leu Glu Asn Val Glu His Ala Tyr 145 150 155 160 Gly Met Arg Ala Ala Gln Phe His Ala Arg Phe Met Asp Ala Ile Thr 165 170 175 Pro Thr Gly Thr Val Ile Thr Leu Leu Gly Leu Thr Pro Glu Gly His 180 185 190 Arg Val Ala Val His Val Tyr Gly Thr Arg Gln Tyr Phe Tyr Met Asn 195 200 205 Lys Glu Glu Val Asp Arg His Leu Gln Cys Arg Ala Pro Arg Asp Leu 210 215 220 Cys Glu Arg Met Ala Ala Ala Leu Arg Glu Ser Pro Gly Ala Ser Phe 225 230 235 240 Arg Gly Ile Ser Ala Asp His Phe Glu Ala Glu Val Val Glu Arg Thr 245 250 255 Asp Val Tyr Tyr Tyr Glu Thr Arg Pro Ala Leu Phe Tyr Arg Val Tyr 260 265 270 Val Arg Ser Gly Arg Val Leu Ser Tyr Leu Cys Asp Asn Phe Cys Pro 275 280 285 Ala Ile Lys Lys Tyr Glu Gly Gly Val Asp Ala Thr Thr Arg Phe Ile 290 295 300 Leu Asp Asn Pro Gly Phe Val Thr Phe Gly Trp Tyr Arg Leu Lys Pro 305 310 315 320 Gly Arg Asn Asn Thr Leu Ala Gln Pro Arg Ala Pro Met Ala Phe Gly 325 330 335 Thr Ser Ser Asp Val Glu Phe Asn Cys Thr Ala Asp Asn Leu Ala Ile 340 345 350 Glu Gly Gly Met Ser Asp Leu Pro Ala Tyr Lys Leu Met Cys Phe Asp 355 360 365 Ile Glu Cys Lys Ala Gly Gly Glu Asp Glu Leu Ala Phe Pro Val Ala 370 375 380 Gly His Pro Glu Asp Leu Val Ile Gln Ile Ser Cys Leu Leu Tyr Asp 385 390 395 400 Leu Ser Thr Thr Ala Leu Glu His Val Leu Leu Phe Ser Leu Gly Ser 405 410 415 Cys Asp Leu Pro Glu Ser His Leu Asn Glu Leu Ala Ala Arg Gly Leu 420 425 430 Pro Thr Pro Val Val Leu Glu Phe Asp Ser Glu Phe Glu Met Leu Leu 435 440 445 Ala Phe Met Thr Leu Val Lys Gln Tyr Gly Pro Glu Phe Val Thr Gly 450 455 460 Tyr Asn Ile Ile Asn Phe Asp Trp Pro Phe Leu Leu Ala Lys Leu Thr 465 470 475 480 Asp Ile Tyr Lys Val Pro Leu Asp Gly Tyr Gly Arg Met Asn Gly Arg 485 490 495 Gly Val Phe Arg Val Trp Asp Ile Gly Gln Ser His Phe Gln Lys Arg 500 505 510 Ser Lys Ile Lys Val Asn Gly Met Val Asn Ile Asp Met Tyr Gly Ile 515 520 525 Ile Thr Asp Lys Ile Lys Leu Ser Ser Tyr Lys Leu Asn Ala Val Ala 530 535 540 Glu Ala Val Leu Lys Asp Lys Lys Lys Asp Leu Ser Tyr Arg Asp Ile 545 550 555 560 Pro Ala Tyr Tyr Ala Ala Gly Pro Ala Gln Arg Gly Val Ile Gly Glu 565 570 575 Tyr Cys Ile Gln Asp Ser Leu Leu Val Gly Gln Leu Phe Phe Lys Phe 580 585 590 Leu Pro His Leu Glu Leu Ser Ala Val Ala Arg Leu Ala Gly Ile Asn 595 600 605 Ile Thr Arg Thr Ile Tyr Asp Gly Gln Gln Ile Arg Val Phe Thr Cys 610 615 620 Leu Leu Arg Leu Ala Asp Gln Lys Gly Phe Ile Leu Pro Asp Thr Gln 625 630 635 640 Gly Arg Phe Arg Gly Gly Gly Gly Glu Ala Pro Lys Arg Pro Ala Ala 645 650 655 Ala Arg Glu Asp Glu Glu Arg Pro Glu Glu Glu Gly Glu Asp Glu Asp 660 665 670 Glu Arg Glu Glu Gly Gly Gly Glu Arg Glu Pro Glu Gly Ala Arg Glu 675 680 685 Thr Ala Gly Arg His Val Gly Tyr Gln Gly Ala Arg Val Leu Asp Pro 690 695 700 Thr Ser Gly Phe His Val Asn Pro Val Val Val Phe Asp Phe Ala Ser 705 710 715 720 Leu Tyr Pro Ser Ile Ile Gln Ala His Asn Leu Cys Phe Ser Thr Leu 725 730 735 Ser Leu Arg Ala Asp Ala Val Ala His Leu Glu Ala Gly Lys Asp Tyr 740 745 750 Leu Glu Ile Glu Val Gly Gly Arg Arg Leu Phe Phe Val Lys Ala His 755 760 765 Val Arg Glu Ser Leu Leu Ser Ile Leu Leu Arg Asp Trp Leu Ala Met 770 775 780 Arg Lys Gln Ile Arg Ser Arg Ile Pro Gln Ser Ser Pro Glu Glu Ala 785 790 795 800 Val Leu Leu Asp Lys Gln Gln Ala Ala Ile Lys Val Val Cys Asn Ser 805 810 815 Val Tyr Gly Phe Thr Gly Ala Gln His Gly Leu Leu Pro Cys Leu His 820 825 830 Val Ala Ala Thr Val Thr Thr Ile Gly Arg Glu Met Leu Leu Ala Thr 835 840 845 Arg Glu Tyr Val His Ala Arg Trp Ala Ala Phe Glu Gln Leu Leu Ala 850 855 860 Asp Phe Pro Glu Ala Ala Asp Met Arg Ala Pro Gly Pro Tyr Ser Met 865 870 875 880 Arg Ile Ile Tyr Gly Asp Thr Asp Ser Ile Phe Val Leu Cys Arg Gly 885 890 895 Leu Thr Ala Ala Gly Leu Thr Ala Val Gly Asp Lys Met Ala Ser His 900 905 910 Ile Ser Arg Ala Leu Phe Leu Ser Pro Ile Lys Leu Glu Cys Glu Lys 915 920 925 Thr Phe Thr Lys Leu Leu Leu Ile Ala Lys Lys Lys Tyr Ile Gly Val 930 935 940 Ile Tyr Gly Gly Lys Met Leu Ile Lys Gly Val Asp Leu Val Arg Lys 945 950 955 960 Asn Asn Cys Ala Phe Ile Asn Arg Thr Ser Arg Ala Leu Val Asp Leu 965 970 975 Leu Phe Tyr Asp Asp Thr Val Ser Gly Ala Ala Ala Ala Leu Ala Glu 980 985 990 Arg Pro Ala Glu Glu Trp Leu Ala Arg Pro Leu Pro Glu Gly Leu Gln 995 1000 1005 Ala Phe Gly Ala Val Leu Val Asp Ala His Arg Arg Ile Thr Asp 1010 1015 1020 Pro Glu Arg Asp Ile Gln Asp Phe Val Leu Thr Ala Glu Leu Ser 1025 1030 1035 Arg His Pro Arg Ala Tyr Thr Asn Lys Arg Leu Ala His Leu Thr 1040 1045 1050 Val Tyr Tyr Lys Leu Met Ala Arg Arg Ala Gln Val Pro Ser Ile 1055 1060 1065 Lys Asp Arg Ile Pro Tyr Val Ile Val Ala Gln Thr Arg Glu Val 1070 1075 1080 Glu Glu Thr Val Ala Arg Leu Ala Ala Leu Arg Glu Leu Asp Ala 1085 1090 1095 Ala Ala Pro Gly Asp Glu Pro Ala Pro Pro Ala Ala Leu Pro Ser 1100 1105 1110 Pro Ala Lys Arg Pro Arg Glu Thr Pro Leu His Ala Asp Pro Pro 1115 1120 1125 Gly Gly Ala Ser Lys Pro Arg Lys Leu Leu Val Ser Glu Leu Ala 1130 1135 1140 Glu Asp Pro Ala Tyr Ala Ile Ala His Gly Val Ala Leu Asn Thr 1145 1150 1155 Asp Tyr Tyr Phe Ser His Leu Leu Gly Ala Ala Cys Val Thr Phe 1160 1165 1170 Lys Ala Leu Phe Gly Asn Asn Ala Lys Ile Thr Glu Ser Leu Leu 1175 1180 1185 Lys Arg Phe Ile Pro Glu Val Trp His Pro Pro Asp Asp Val Ala 1190 1195 1200 Ala Arg Leu Arg Ala Ala Gly Phe Gly Ala Val Gly Ala Gly Ala 1205 1210 1215 Thr Ala Glu Glu Thr Arg Arg Met Leu His Arg Ala Phe Asp Thr 1220 1225 1230 Leu Ala 1235 9 3708 DNA herpes simplex 9 atgttttccg gtggcggcgg cccgctgtcc cccggaggaa agtcggcggc cagggcggcg 60 tccgggtttt ttgcgcccgc cggccctcgc ggagccggcc ggggaccccc gccttgtttg 120 aggcaaaact tttacaaccc ctacctcgcc ccagtcggga cgcaacagaa gccgaccggg 180 ccaacccagc gccatacgta ctatagcgaa tgcgatgaat ttcgattcat cgccccgcgg 240 gtgctggacg aggatgcccc cccggagaag cgcgccgggg tgcacgacgg tcacctcaag 300 cgcgccccca aggtgtactg cgggggggac gagcgcgacg tcctccgcgt cgggtcgggc 360 ggcttctggc cgcggcgctc gcgcctgtgg ggcggcgtgg accacgcccc ggcggggttc 420 aaccccaccg tcaccgtctt tcacgtgtat gacatcctgg agaacgtgga gcacgcgtac 480 ggcatgcgcg cggcccagtt ccacgcgcgg tttatggacg ccatcacacc gacggggacc 540 gtcatcacgc tcctgggcct gactccggaa ggccaccggg tggccgttca cgtttacggc 600 acgcggcagt acttttacat gaacaaggag gaggttgaca ggcacctaca atgccgcgcc 660 ccacgagatc tctgcgagcg catggccgcg gccctgcgcg agtccccggg cgcgtcgttc 720 cgcggcatct ccgcggacca cttcgaggcg gaggtggtgg agcgcaccga cgtgtactac 780 tacgagacgc gccccgctct gttttaccgc gtctacgtcc gaagcgggcg cgtgctgtcg 840 tacctgtgcg acaacttctg cccggccatc aagaagtacg agggtggggt cgacgccacc 900 acccggttca tcctggacaa ccccgggttc gtcaccttcg gctggtaccg tctcaaaccg 960 ggccggaaca acacgctagc ccagccgcgg gccccgatgg ccttcgggac atccagcgat 1020 gtcgagttta actgtacggc ggacaacctg gccatcgagg ggggcatgag cgacctaccg 1080 gcatacaagc tcatgtgctt cgatatcgaa tgcaaggcgg ggggggagga cgagctggcc 1140 tttccggtgg ccgggcaccc ggaggacctg gtcatccaga tatcctgtct gctctacgac 1200 ctgtccacca ccgccctgga gcacgtcctc ctgttttcgc tcggttcctg cgacctcccc 1260 gaatcccacc tgaacgagct ggcggccagg ggcctgccca cgcccgtggt tctggaattc 1320 gacagcgaat tcgagatgct gttggccttc atgacccttg tgaaacagta cggccccgag 1380 ttcgtgaccg ggtacaacat aatcaacttc gactggccct tcttgctggc caagctgacg 1440 gacatttaca aggtccccct ggacgggtac ggccgcatga acggccgggg cgtgtttcgc 1500 gtgtgggaca taggccagag ccacttccag aagcgcagca agataaaggt gaacggcatg 1560 gtgaacatcg acatgtacgg gattataacc gacaagatca agctctcgag ctacaagctc 1620 aacgccgtgg ccgaagccgt cctgaaggac aagaagaagg acctgagcta tcgcgacatc 1680 cccacctact acgccgccgg gcccgcgcaa cgcggggtga tcggcgagta ctgcatacag 1740 gattccctgc tggtgggcca gctgtttttt aagtttttgc cccatctgga gctctcggcc 1800 gtcgcgcgct tggcgggtat taacatcacc cgcaccatct acgacggcca gcagatccgc 1860 gtctttacgt gcctgctgcg cctggccgac cagaagggct ttattctgcc ggacacccag 1920 gggcgattta ggggcgccgg gggggaggcg cccaagcgtc cggccgcagc ccgggaggac 1980 gaggagcggc cagaggagga gggggaggac gagaacgaac gcgaggaggg cgggggcgag 2040 cgggagccgg agggcgcgcg ggagaccgcc ggccggcacg tggggtacca gggggccagg 2100 gtccttgacc ccacttccgg gtttcacgtg aaccccgtgg tggtgttcga ctttgccagc 2160 ctgtacccca gcatcatcca ggcccacaac ctgtgcttca gcacgctctc cctgagggcc 2220 gacgcagtgg cgcacctgga ggcgggcaag gactacctgg agatcgaggt gggggggcga 2280 cggctgttct tcgtcaaggc tcacgtgcga gagagcctcc tcagcatcct cctgcgggac 2340 tggctcgcca tgcgaaagca gatccgctcg cggattcccc agagcagccc cgaggaggcc 2400 gtgctcctgg acaagcagca ggccgccatc aaggtcgtgt gtaactcggt ttacgggttc 2460 acgggagcgc agcacggact cctgccgtgc ctgcacgttg ccgcgacggt gacgaccatc 2520 ggccgcgaga tgctgctcgc gacccgcgag tacgtccacg cgcgctgggc ggccttcgaa 2580 cagctcctgg ccgatttccc ggaggcggcc gacatgcgcg cccccgggcc ctattccatg 2640 cgcatcatct acggggacac ggactccata tttgtgctgt gccgcggcct cacggccgcc 2700 gggctgacgg ccgtgggcga caagatggcg agccacatct cgcgcgcgct gtttctgccc 2760 cccatcaaac tcgagtgcga aaagacgttc accaagctgc tgctgatcgc caagaaaaag 2820 tacatcggcg tcatctacgg gggtaagatg ctcatcaagg gcgtggatct ggtgcgcaaa 2880 aacaactgcg cgtttatcaa ccgcacctcc agggccctgg tcgacctgct gttttacgac 2940 gataccgtat ccggagcggc cgccgcgtta gccgagcgcc ccgcagagga gtggctggcg 3000 cgacccctgc ccgagggact gcaggcgttc ggggccgtcc tcgtagacgc ccatcggcgc 3060 atcaccgacc cggagaggga catccaggac tttgttctca ccgccgaact gagcagacac 3120 ccgcgcgcgt acaccaacaa gcgcctggcc cacctgacgg tgtattacaa gctcatggcc 3180 cgccgcgcgc aggtcccgtc catcaaggac cggatcccgt acgtgatcgt ggcccagacc 3240 cgcgaggtag aggagacggt cgcgcggctg gccgccctcc gcgagctaga cgccgccgcc 3300 ccaggggacg agcccgcccc ccccgcggcc ctgccctccc cggccaagcg cccccgggag 3360 acgccgtcgc ctgccgaccc cccgggaggc gcgtccaagc cccgcaagct gctggtgtcc 3420 gagctggccg aggatcccgc atacgccatt gcccacggcg tcgccctgaa cacggactat 3480 tacttctccc acctgttggg ggcggcgtgc gtgacattca aggccctgtt tgggaataac 3540 gccaagatca ccgagagtct gttaaaaagg tttattcccg aagtgtggca ccccccggac 3600 gacgtggccg cgcggctccg gaccgcaggg ttcggggcgg tgggtgccgg cgctacggcg 3660 gaggaaactc gtcgaatgtt gcatagagcc tttgatactc tagcatga 3708 10 1235 PRT herpes simplex 10 Met Phe Ser Gly Gly Gly Gly Pro Leu Ser Pro Gly Gly Lys Ser Ala 1 5 10 15 Ala Arg Ala Ala Ser Gly Phe Phe Ala Pro Ala Gly Pro Arg Gly Ala 20 25 30 Gly Arg Gly Pro Pro Pro Cys Leu Arg Gln Asn Phe Tyr Asn Pro Tyr 35 40 45 Leu Ala Pro Val Gly Thr Gln Gln Lys Pro Thr Gly Pro Thr Gln Arg 50 55 60 His Thr Tyr Tyr Ser Glu Cys Asp Glu Phe Arg Phe Ile Ala Pro Arg 65 70 75 80 Val Leu Asp Glu Asp Ala Pro Pro Glu Lys Arg Ala Gly Val His Asp 85 90 95 Gly His Leu Lys Arg Ala Pro Lys Val Tyr Cys Gly Gly Asp Glu Arg 100 105 110 Asp Val Leu Arg Val Gly Ser Gly Gly Phe Trp Pro Arg Arg Ser Arg 115 120 125 Leu Trp Gly Gly Val Asp His Ala Pro Ala Gly Phe Asn Pro Thr Val 130 135 140 Thr Val Phe His Val Tyr Asp Ile Leu Glu Asn Val Glu His Ala Tyr 145 150 155 160 Gly Met Arg Ala Ala Gln Phe His Ala Arg Phe Met Asp Ala Ile Thr 165 170 175 Pro Thr Gly Thr Val Ile Thr Leu Leu Gly Leu Thr Pro Glu Gly His 180 185 190 Arg Val Ala Val His Val Tyr Gly Thr Arg Gln Tyr Phe Tyr Met Asn 195 200 205 Lys Glu Glu Val Asp Arg His Leu Gln Cys Arg Ala Pro Arg Asp Leu 210 215 220 Cys Glu Arg Met Ala Ala Ala Leu Arg Glu Ser Pro Gly Ala Ser Phe 225 230 235 240 Arg Gly Ile Ser Ala Asp His Phe Glu Ala Glu Val Val Glu Arg Thr 245 250 255 Asp Val Tyr Tyr Tyr Glu Thr Arg Pro Ala Leu Phe Tyr Arg Val Tyr 260 265 270 Val Arg Ser Gly Arg Val Leu Ser Tyr Leu Cys Asp Asn Phe Cys Pro 275 280 285 Ala Ile Lys Lys Tyr Glu Gly Gly Val Asp Ala Thr Thr Arg Phe Ile 290 295 300 Leu Asp Asn Pro Gly Phe Val Thr Phe Gly Trp Tyr Arg Leu Lys Pro 305 310 315 320 Gly Arg Asn Asn Thr Leu Ala Gln Pro Arg Ala Pro Met Ala Phe Gly 325 330 335 Thr Ser Ser Asp Val Glu Phe Asn Cys Thr Ala Asp Asn Leu Ala Ile 340 345 350 Glu Gly Gly Met Ser Asp Leu Pro Ala Tyr Lys Leu Met Cys Phe Asp 355 360 365 Ile Glu Cys Lys Ala Gly Gly Glu Asp Glu Leu Ala Phe Pro Val Ala 370 375 380 Gly His Pro Glu Asp Leu Val Ile Gln Ile Ser Cys Leu Leu Tyr Asp 385 390 395 400 Leu Ser Thr Thr Ala Leu Glu His Val Leu Leu Phe Ser Leu Gly Ser 405 410 415 Cys Asp Leu Pro Glu Ser His Leu Asn Glu Leu Ala Ala Arg Gly Leu 420 425 430 Pro Thr Pro Val Val Leu Glu Phe Asp Ser Glu Phe Glu Met Leu Leu 435 440 445 Ala Phe Met Thr Leu Val Lys Gln Tyr Gly Pro Glu Phe Val Thr Gly 450 455 460 Tyr Asn Ile Ile Asn Phe Asp Trp Pro Phe Leu Leu Ala Lys Leu Thr 465 470 475 480 Asp Ile Tyr Lys Val Pro Leu Asp Gly Tyr Gly Arg Met Asn Gly Arg 485 490 495 Gly Val Phe Arg Val Trp Asp Ile Gly Gln Ser His Phe Gln Lys Arg 500 505 510 Ser Lys Ile Lys Val Asn Gly Met Val Asn Ile Asp Met Tyr Gly Ile 515 520 525 Ile Thr Asp Lys Ile Lys Leu Ser Ser Tyr Lys Leu Asn Ala Val Ala 530 535 540 Glu Ala Val Leu Lys Asp Lys Lys Lys Asp Leu Ser Tyr Arg Asp Ile 545 550 555 560 Pro Thr Tyr Tyr Ala Ala Gly Pro Ala Gln Arg Gly Val Ile Gly Glu 565 570 575 Tyr Cys Ile Gln Asp Ser Leu Leu Val Gly Gln Leu Phe Phe Lys Phe 580 585 590 Leu Pro His Leu Glu Leu Ser Ala Val Ala Arg Leu Ala Gly Ile Asn 595 600 605 Ile Thr Arg Thr Ile Tyr Asp Gly Gln Gln Ile Arg Val Phe Thr Cys 610 615 620 Leu Leu Arg Leu Ala Asp Gln Lys Gly Phe Ile Leu Pro Asp Thr Gln 625 630 635 640 Gly Arg Phe Arg Gly Ala Gly Gly Glu Ala Pro Lys Arg Pro Ala Ala 645 650 655 Ala Arg Glu Asp Glu Glu Arg Pro Glu Glu Glu Gly Glu Asp Glu Asn 660 665 670 Glu Arg Glu Glu Gly Gly Gly Glu Arg Glu Pro Glu Gly Ala Arg Glu 675 680 685 Thr Ala Gly Arg His Val Gly Tyr Gln Gly Ala Arg Val Leu Asp Pro 690 695 700 Thr Ser Gly Phe His Val Asn Pro Val Val Val Phe Asp Phe Ala Ser 705 710 715 720 Leu Tyr Pro Ser Ile Ile Gln Ala His Asn Leu Cys Phe Ser Thr Leu 725 730 735 Ser Leu Arg Ala Asp Ala Val Ala His Leu Glu Ala Gly Lys Asp Tyr 740 745 750 Leu Glu Ile Glu Val Gly Gly Arg Arg Leu Phe Phe Val Lys Ala His 755 760 765 Val Arg Glu Ser Leu Leu Ser Ile Leu Leu Arg Asp Trp Leu Ala Met 770 775 780 Arg Lys Gln Ile Arg Ser Arg Ile Pro Gln Ser Ser Pro Glu Glu Ala 785 790 795 800 Val Leu Leu Asp Lys Gln Gln Ala Ala Ile Lys Val Val Cys Asn Ser 805 810 815 Val Tyr Gly Phe Thr Gly Ala Gln His Gly Leu Leu Pro Cys Leu His 820 825 830 Val Ala Ala Thr Val Thr Thr Ile Gly Arg Glu Met Leu Leu Ala Thr 835 840 845 Arg Glu Tyr Val His Ala Arg Trp Ala Ala Phe Glu Gln Leu Leu Ala 850 855 860 Asp Phe Pro Glu Ala Ala Asp Met Arg Ala Pro Gly Pro Tyr Ser Met 865 870 875 880 Arg Ile Ile Tyr Gly Asp Thr Asp Ser Ile Phe Val Leu Cys Arg Gly 885 890 895 Leu Thr Ala Ala Gly Leu Thr Ala Val Gly Asp Lys Met Ala Ser His 900 905 910 Ile Ser Arg Ala Leu Phe Leu Pro Pro Ile Lys Leu Glu Cys Glu Lys 915 920 925 Thr Phe Thr Lys Leu Leu Leu Ile Ala Lys Lys Lys Tyr Ile Gly Val 930 935 940 Ile Tyr Gly Gly Lys Met Leu Ile Lys Gly Val Asp Leu Val Arg Lys 945 950 955 960 Asn Asn Cys Ala Phe Ile Asn Arg Thr Ser Arg Ala Leu Val Asp Leu 965 970 975 Leu Phe Tyr Asp Asp Thr Val Ser Gly Ala Ala Ala Ala Leu Ala Glu 980 985 990 Arg Pro Ala Glu Glu Trp Leu Ala Arg Pro Leu Pro Glu Gly Leu Gln 995 1000 1005 Ala Phe Gly Ala Val Leu Val Asp Ala His Arg Arg Ile Thr Asp 1010 1015 1020 Pro Glu Arg Asp Ile Gln Asp Phe Val Leu Thr Ala Glu Leu Ser 1025 1030 1035 Arg His Pro Arg Ala Tyr Thr Asn Lys Arg Leu Ala His Leu Thr 1040 1045 1050 Val Tyr Tyr Lys Leu Met Ala Arg Arg Ala Gln Val Pro Ser Ile 1055 1060 1065 Lys Asp Arg Ile Pro Tyr Val Ile Val Ala Gln Thr Arg Glu Val 1070 1075 1080 Glu Glu Thr Val Ala Arg Leu Ala Ala Leu Arg Glu Leu Asp Ala 1085 1090 1095 Ala Ala Pro Gly Asp Glu Pro Ala Pro Pro Ala Ala Leu Pro Ser 1100 1105 1110 Pro Ala Lys Arg Pro Arg Glu Thr Pro Ser Pro Ala Asp Pro Pro 1115 1120 1125 Gly Gly Ala Ser Lys Pro Arg Lys Leu Leu Val Ser Glu Leu Ala 1130 1135 1140 Glu Asp Pro Ala Tyr Ala Ile Ala His Gly Val Ala Leu Asn Thr 1145 1150 1155 Asp Tyr Tyr Phe Ser His Leu Leu Gly Ala Ala Cys Val Thr Phe 1160 1165 1170 Lys Ala Leu Phe Gly Asn Asn Ala Lys Ile Thr Glu Ser Leu Leu 1175 1180 1185 Lys Arg Phe Ile Pro Glu Val Trp His Pro Pro Asp Asp Val Ala 1190 1195 1200 Ala Arg Leu Arg Thr Ala Gly Phe Gly Ala Val Gly Ala Gly Ala 1205 1210 1215 Thr Ala Glu Glu Thr Arg Arg Met Leu His Arg Ala Phe Asp Thr 1220 1225 1230 Leu Ala 1235 11 3729 DNA herpes simplex 11 atgtttttca acccgtatct gagcggcggc gtgaccggcg gtgcggtcgc gggtggccgg 60 cgtcagcgtt cgcagcccgg ctccgcgcag ggctcgggca agcggccgcc acagaaacag 120 tttttgcaga tcgtgccgcg aggtgtcatg ttcgacggtc agacggggtt gatcaagcat 180 aagacgggac ggctgcctct catgttctat cgagagatta aacatttgtt gagtcatgac 240 atggtttggc cgtgtccttg gcgcgagacc ctggtgggtc gcgtggtggg acctattcgt 300 tttcacacct acgatcagac ggacgccgtg ctcttcttcg actcgcccga aaacgtgtcg 360 ccgcgctatc gtcagcatct ggtgccttcg gggaacgtgt tgcgtttctt cggggccaca 420 gaacacggct acagtatctg cgtcaacgtt ttcgggcagc gcagctactt ttactgtgag 480 tacagcgaca ccgataggct gcgtgaggtc attgccagcg tgggcgaact agtgcccgaa 540 ccgcggacgc catacgccgt gtctgtcacg ccggccacca agacctccat ctatgggtac 600 gggacgcgac ccgtgcccga tttgcagtgt gtgtctatca gcaactggac catggccaga 660 aaaatcggcg agtatctgct ggagcagggt tttcccgtgt acgaggtccg tgtggatccg 720 ctgacgcgtt tggtcatcga tcggcggatc accacgttcg gctggtgctc cgtgaatcgt 780 tacgactggc ggcagcaggg tcgcgcgtcg acttgtgata tcgaggtaga ctgcgatgtc 840 tctgacctgg tggctgtgcc cgacgacagc tcgtggccgc gctatcgatg cctgtccttc 900 gatatcgagt gcatgagcgg cgagggtggt tttccctgcg ccgagaagtc cgatgacatt 960 gtcattcaga tctcgtgcgt gtgctacgag acggggggaa acaccgccgt ggatcagggg 1020 atcccaaacg ggaacgatgg tcggggctgc acttcggagg gtgtgatctt tgggcactcg 1080 ggtcttcatc tctttacgat cggcacctgc gggcaggtgg gcccagacgt ggacgtctac 1140 gagttccctt ccgaatacga gctgctgctg ggctttatgc ttttctttca acggtacgcg 1200 ccggcctttg tgaccggtta caacatcaac tcttttgact tgaagtacat cctcacgcgt 1260 ctcgagtacc tgtataaggt ggactcgcag cgcttctgca agttgcctac ggcgcagggc 1320 ggccgtttct ttttacacag ccccgccgtg ggttttaagc ggcagtacgc cgccgctttt 1380 ccctcggctt ctcacaacaa tccggccagc acggccgcca ccaaggtgta tattgcgggt 1440 tcggtggtta tcgacatgta ccctgtatgc atggccaaga ctaactcgcc caactataag 1500 ctcaacacta tggccgagct ttacctgcgg caacgcaagg atgacctgtc ttacaaggac 1560 atcccgcgtt gtttcgtggc taatgccgag ggccgcgccc aggtaggccg ttactgtctg 1620 caggacgccg tattggtgcg cgatctgttc aacaccatta attttcacta cgaggccggg 1680 gccatcgcgc ggctggctaa aattccgttg cggcgtgtca tctttgacgg acagcagatc 1740 cgtatctaca cctcgctgct ggacgagtgc gcctgccgcg attttatcct gcccaaccac 1800 tacagcaaag gtacgacggt gcccgaaacg aatagcgttg ctgtgtcacc taacgctgct 1860 atcatctcta ccgccgctgt gcccggcgac gcgggttctg tggcggctat gtttcagatg 1920 tcgccgccct tgcaatctgc gccgtccagt caggacggcg tttcacccgg ctccggcagt 1980 aacagtagta gcagcgtcgg cgttttcagc gtcggctccg gcagtagtgg cggcgtcggc 2040 gtttccaacg acaatcacgg cgccggcggt actgcggcgg tttcgtacca gggcgccacg 2100 gtgtttgagc ccgaggtggg ttactacaac gaccccgtgg ccgtgttcga ctttgccagc 2160 ctctaccctt ccatcatcat ggcccacaac ctctgctact ccaccctgct ggtgccgggt 2220 ggcgagtacc ctgtggaccc cgccgacgta tacagcgtca cgctagagaa cggcgtgacc 2280 caccgctttg tgcgtgcttc ggtgcgcgtc tcggtgctct cggaactgct caacaagtgg 2340 gtttcgcagc ggcgtgccgt gcgcgaatgc atgcgcgagt gtcaagaccc tgtgcgccgt 2400 atgctgctcg acaaggaaca gatggcgctc aaagtaacgt gcaacgcttt ctacggtttt 2460 accggcgcgc tgaacggtat gatgccgtgt ctgcccatcg ccgccagcat cacgcgcatc 2520 ggtcgcgaca tgctagagcg cacggcgcgg ttcatcaaag acaacttttc agagccgtgt 2580 tttttgcaca atttttttaa tcaggaagac tatgtagtgg gaacgcggga gggggattcg 2640 gaggagagca gcgcgttacc ggaggggctc gaaacatcgt cagggggctc gaacgaacgg 2700 cgggtggagg cgcgggtcat ctacggggac acggacagcg tgtttgtccg ctttcgtggc 2760 ctgacgccgc aggctctggt ggcgcgtggg cccagcctgg cgcactacgt gacggcctgt 2820 ctttttgtgg agcccgtcaa gctggagttt gaaaaggtct tcgtctctct tatgatgatc 2880 tgcaagaaac gttacatcgg caaagtggag ggcgcctcgg gtctgagcat gaagggcgtg 2940 gatctggtgc gcaagacggc ctgcgagttc gtcaagggcg tcacgcgtga cgtcctctcg 3000 ctgctctttg aggatcgcga ggtctcggaa gcagccgtgc gcctgtcgcg cctctcactc 3060 gatgaagtca agaagtacgg cgtgccacgc ggtttctggc gtatcttacg ccgcttggtg 3120 caggcccgcg acgatctgta cctgcaccgt gtgcgtgtcg aggacctggt gctttcgtcg 3180 gtgctctcta aggacatctc gctgtaccgt caatctaacc tgccgcacat tgccgtcatt 3240 aagcgattgg cggcccgttc tgaggagcta ccctcggtcg gggatcgggt cttttacgtt 3300 ctgacggcgc ccggtgtccg gacggcgccg cagggttcct ccgacaacgg tgattctgta 3360 accgccggcg tggtttcccg gtcggacgcg attgatggca cggacgacga cgctgacggc 3420 ggcggggtag aggagagcaa caggagagga ggagagccgg caaagaagag ggcgcggaaa 3480 ccaccgtcgg ccgtgtgcaa ctacgaggta gccgaagatc cgagctacgt gcgcgagcac 3540 ggcgtgccca ttcacgccga caagtacttt gagcaggttc tcaaggctgt aactaacgtg 3600 ctgtcgcccg tctttcccgg cggcgaaacc gcgcgcaagg acaagttttt gcacatggtg 3660 ctgccgcggc gcttgcactt ggagccggct tttctgccgt acagtgtcaa ggcgcacgaa 3720 tgctgttga 3729 12 1242 PRT herpes simplex 12 Met Phe Phe Asn Pro Tyr Leu Ser Gly Gly Val Thr Gly Gly Ala Val 1 5 10 15 Ala Gly Gly Arg Arg Gln Arg Ser Gln Pro Gly Ser Ala Gln Gly Ser 20 25 30 Gly Lys Arg Pro Pro Gln Lys Gln Phe Leu Gln Ile Val Pro Arg Gly 35 40 45 Val Met Phe Asp Gly Gln Thr Gly Leu Ile Lys His Lys Thr Gly Arg 50 55 60 Leu Pro Leu Met Phe Tyr Arg Glu Ile Lys His Leu Leu Ser His Asp 65 70 75 80 Met Val Trp Pro Cys Pro Trp Arg Glu Thr Leu Val Gly Arg Val Val 85 90 95 Gly Pro Ile Arg Phe His Thr Tyr Asp Gln Thr Asp Ala Val Leu Phe 100 105 110 Phe Asp Ser Pro Glu Asn Val Ser Pro Arg Tyr Arg Gln His Leu Val 115 120 125 Pro Ser Gly Asn Val Leu Arg Phe Phe Gly Ala Thr Glu His Gly Tyr 130 135 140 Ser Ile Cys Val Asn Val Phe Gly Gln Arg Ser Tyr Phe Tyr Cys Glu 145 150 155 160 Tyr Ser Asp Thr Asp Arg Leu Arg Glu Val Ile Ala Ser Val Gly Glu 165 170 175 Leu Val Pro Glu Pro Arg Thr Pro Tyr Ala Val Ser Val Thr Pro Ala 180 185 190 Thr Lys Thr Ser Ile Tyr Gly Tyr Gly Thr Arg Pro Val Pro Asp Leu 195 200 205 Gln Cys Val Ser Ile Ser Asn Trp Thr Met Ala Arg Lys Ile Gly Glu 210 215 220 Tyr Leu Leu Glu Gln Gly Phe Pro Val Tyr Glu Val Arg Val Asp Pro 225 230 235 240 Leu Thr Arg Leu Val Ile Asp Arg Arg Ile Thr Thr Phe Gly Trp Cys 245 250 255 Ser Val Asn Arg Tyr Asp Trp Arg Gln Gln Gly Arg Ala Ser Thr Cys 260 265 270 Asp Ile Glu Val Asp Cys Asp Val Ser Asp Leu Val Ala Val Pro Asp 275 280 285 Asp Ser Ser Trp Pro Arg Tyr Arg Cys Leu Ser Phe Asp Ile Glu Cys 290 295 300 Met Ser Gly Glu Gly Gly Phe Pro Cys Ala Glu Lys Ser Asp Asp Ile 305 310 315 320 Val Ile Gln Ile Ser Cys Val Cys Tyr Glu Thr Gly Gly Asn Thr Ala 325 330 335 Val Asp Gln Gly Ile Pro Asn Gly Asn Asp Gly Arg Gly Cys Thr Ser 340 345 350 Glu Gly Val Ile Phe Gly His Ser Gly Leu His Leu Phe Thr Ile Gly 355 360 365 Thr Cys Gly Gln Val Gly Pro Asp Val Asp Val Tyr Glu Phe Pro Ser 370 375 380 Glu Tyr Glu Leu Leu Leu Gly Phe Met Leu Phe Phe Gln Arg Tyr Ala 385 390 395 400 Pro Ala Phe Val Thr Gly Tyr Asn Ile Asn Ser Phe Asp Leu Lys Tyr 405 410 415 Ile Leu Thr Arg Leu Glu Tyr Leu Tyr Lys Val Asp Ser Gln Arg Phe 420 425 430 Cys Lys Leu Pro Thr Ala Gln Gly Gly Arg Phe Phe Leu His Ser Pro 435 440 445 Ala Val Gly Phe Lys Arg Gln Tyr Ala Ala Ala Phe Pro Ser Ala Ser 450 455 460 His Asn Asn Pro Ala Ser Thr Ala Ala Thr Lys Val Tyr Ile Ala Gly 465 470 475 480 Ser Val Val Ile Asp Met Tyr Pro Val Cys Met Ala Lys Thr Asn Ser 485 490 495 Pro Asn Tyr Lys Leu Asn Thr Met Ala Glu Leu Tyr Leu Arg Gln Arg 500 505 510 Lys Asp Asp Leu Ser Tyr Lys Asp Ile Pro Arg Cys Phe Val Ala Asn 515 520 525 Ala Glu Gly Arg Ala Gln Val Gly Arg Tyr Cys Leu Gln Asp Ala Val 530 535 540 Leu Val Arg Asp Leu Phe Asn Thr Ile Asn Phe His Tyr Glu Ala Gly 545 550 555 560 Ala Ile Ala Arg Leu Ala Lys Ile Pro Leu Arg Arg Val Ile Phe Asp 565 570 575 Gly Gln Gln Ile Arg Ile Tyr Thr Ser Leu Leu Asp Glu Cys Ala Cys 580 585 590 Arg Asp Phe Ile Leu Pro Asn His Tyr Ser Lys Gly Thr Thr Val Pro 595 600 605 Glu Thr Asn Ser Val Ala Val Ser Pro Asn Ala Ala Ile Ile Ser Thr 610 615 620 Ala Ala Val Pro Gly Asp Ala Gly Ser Val Ala Ala Met Phe Gln Met 625 630 635 640 Ser Pro Pro Leu Gln Ser Ala Pro Ser Ser Gln Asp Gly Val Ser Pro 645 650 655 Gly Ser Gly Ser Asn Ser Ser Ser Ser Val Gly Val Phe Ser Val Gly 660 665 670 Ser Gly Ser Ser Gly Gly Val Gly Val Ser Asn Asp Asn His Gly Ala 675 680 685 Gly Gly Thr Ala Ala Val Ser Tyr Gln Gly Ala Thr Val Phe Glu Pro 690 695 700 Glu Val Gly Tyr Tyr Asn Asp Pro Val Ala Val Phe Asp Phe Ala Ser 705 710 715 720 Leu Tyr Pro Ser Ile Ile Met Ala His Asn Leu Cys Tyr Ser Thr Leu 725 730 735 Leu Val Pro Gly Gly Glu Tyr Pro Val Asp Pro Ala Asp Val Tyr Ser 740 745 750 Val Thr Leu Glu Asn Gly Val Thr His Arg Phe Val Arg Ala Ser Val 755 760 765 Arg Val Ser Val Leu Ser Glu Leu Leu Asn Lys Trp Val Ser Gln Arg 770 775 780 Arg Ala Val Arg Glu Cys Met Arg Glu Cys Gln Asp Pro Val Arg Arg 785 790 795 800 Met Leu Leu Asp Lys Glu Gln Met Ala Leu Lys Val Thr Cys Asn Ala 805 810 815 Phe Tyr Gly Phe Thr Gly Ala Leu Asn Gly Met Met Pro Cys Leu Pro 820 825 830 Ile Ala Ala Ser Ile Thr Arg Ile Gly Arg Asp Met Leu Glu Arg Thr 835 840 845 Ala Arg Phe Ile Lys Asp Asn Phe Ser Glu Pro Cys Phe Leu His Asn 850 855 860 Phe Phe Asn Gln Glu Asp Tyr Val Val Gly Thr Arg Glu Gly Asp Ser 865 870 875 880 Glu Glu Ser Ser Ala Leu Pro Glu Gly Leu Glu Thr Ser Ser Gly Gly 885 890 895 Ser Asn Glu Arg Arg Val Glu Ala Arg Val Ile Tyr Gly Asp Thr Asp 900 905 910 Ser Val Phe Val Arg Phe Arg Gly Leu Thr Pro Gln Ala Leu Val Ala 915 920 925 Arg Gly Pro Ser Leu Ala His Tyr Val Thr Ala Cys Leu Phe Val Glu 930 935 940 Pro Val Lys Leu Glu Phe Glu Lys Val Phe Val Ser Leu Met Met Ile 945 950 955 960 Cys Lys Lys Arg Tyr Ile Gly Lys Val Glu Gly Ala Ser Gly Leu Ser 965 970 975 Met Lys Gly Val Asp Leu Val Arg Lys Thr Ala Cys Glu Phe Val Lys 980 985 990 Gly Val Thr Arg Asp Val Leu Ser Leu Leu Phe Glu Asp Arg Glu Val 995 1000 1005 Ser Glu Ala Ala Val Arg Leu Ser Arg Leu Ser Leu Asp Glu Val 1010 1015 1020 Lys Lys Tyr Gly Val Pro Arg Gly Phe Trp Arg Ile Leu Arg Arg 1025 1030 1035 Leu Val Gln Ala Arg Asp Asp Leu Tyr Leu His Arg Val Arg Val 1040 1045 1050 Glu Asp Leu Val Leu Ser Ser Val Leu Ser Lys Asp Ile Ser Leu 1055 1060 1065 Tyr Arg Gln Ser Asn Leu Pro His Ile Ala Val Ile Lys Arg Leu 1070 1075 1080 Ala Ala Arg Ser Glu Glu Leu Pro Ser Val Gly Asp Arg Val Phe 1085 1090 1095 Tyr Val Leu Thr Ala Pro Gly Val Arg Thr Ala Pro Gln Gly Ser 1100 1105 1110 Ser Asp Asn Gly Asp Ser Val Thr Ala Gly Val Val Ser Arg Ser 1115 1120 1125 Asp Ala Ile Asp Gly Thr Asp Asp Asp Ala Asp Gly Gly Gly Val 1130 1135 1140 Glu Glu Ser Asn Arg Arg Gly Gly Glu Pro Ala Lys Lys Arg Ala 1145 1150 1155 Arg Lys Pro Pro Ser Ala Val Cys Asn Tyr Glu Val Ala Glu Asp 1160 1165 1170 Pro Ser Tyr Val Arg Glu His Gly Val Pro Ile His Ala Asp Lys 1175 1180 1185 Tyr Phe Glu Gln Val Leu Lys Ala Val Thr Asn Val Leu Ser Pro 1190 1195 1200 Val Phe Pro Gly Gly Glu Thr Ala Arg Lys Asp Lys Phe Leu His 1205 1210 1215 Met Val Leu Pro Arg Arg Leu His Leu Glu Pro Ala Phe Leu Pro 1220 1225 1230 Tyr Ser Val Lys Ala His Glu Cys Cys 1235 1240 13 1242 PRT herpes simplex 13 Met Phe Phe Asn Pro Tyr Leu Ser Gly Gly Val Thr Gly Gly Ala Val 1 5 10 15 Ala Gly Gly Arg Arg Gln Arg Ser Gln Pro Gly Ser Ala Gln Gly Ser 20 25 30 Gly Lys Arg Pro Pro Gln Lys Gln Phe Leu Gln Ile Val Pro Arg Gly 35 40 45 Val Met Phe Asp Gly Gln Thr Gly Leu Ile Lys His Lys Thr Gly Arg 50 55 60 Leu Pro Leu Met Phe Tyr Arg Glu Ile Lys His Leu Leu Ser His Asp 65 70 75 80 Met Val Trp Pro Cys Pro Trp Arg Glu Thr Leu Val Gly Arg Val Val 85 90 95 Gly Pro Ile Arg Phe His Thr Tyr Asp Gln Thr Asp Ala Val Leu Phe 100 105 110 Phe Asp Ser Pro Glu Asn Val Ser Pro Arg Tyr Arg Gln His Leu Val 115 120 125 Pro Ser Gly Asn Val Leu Arg Phe Phe Gly Ala Thr Glu His Gly Tyr 130 135 140 Ser Ile Cys Val Asn Val Phe Gly Gln Arg Ser Tyr Phe Tyr Cys Glu 145 150 155 160 Tyr Ser Asp Thr Asp Arg Leu Arg Glu Val Ile Ala Ser Val Gly Glu 165 170 175 Leu Val Pro Glu Pro Arg Thr Pro Tyr Ala Val Ser Val Thr Pro Ala 180 185 190 Thr Lys Thr Ser Ile Tyr Gly Tyr Gly Thr Arg Pro Val Pro Asp Leu 195 200 205 Gln Cys Val Ser Ile Ser Asn Trp Thr Met Ala Arg Lys Ile Gly Glu 210 215 220 Tyr Leu Leu Glu Gln Gly Phe Pro Val Tyr Glu Val Arg Val Asp Pro 225 230 235 240 Leu Thr Arg Leu Val Ile Asp Arg Arg Ile Thr Thr Phe Gly Trp Cys 245 250 255 Ser Val Asn Arg Tyr Asp Trp Arg Gln Gln Gly Arg Ala Ser Thr Cys 260 265 270 Asp Ile Glu Val Asp Cys Asp Val Ser Asp Leu Val Ala Val Pro Asp 275 280 285 Asp Ser Ser Trp Pro Arg Tyr Arg Cys Leu Ser Phe Asp Ile Glu Cys 290 295 300 Met Ser Gly Glu Gly Gly Phe Pro Cys Ala Glu Lys Ser Asp Asp Ile 305 310 315 320 Val Ile Gln Ile Ser Cys Val Cys Tyr Glu Thr Gly Gly Asn Thr Ala 325 330 335 Val Asp Gln Gly Ile Pro Asn Gly Asn Asp Gly Arg Gly Cys Thr Ser 340 345 350 Glu Gly Val Ile Phe Gly His Ser Gly Leu His Leu Phe Thr Ile Gly 355 360 365 Thr Cys Gly Gln Val Gly Pro Asp Val Asp Val Tyr Glu Phe Pro Ser 370 375 380 Glu Tyr Glu Leu Leu Leu Gly Phe Met Leu Phe Phe Gln Arg Tyr Ala 385 390 395 400 Pro Ala Phe Val Thr Gly Tyr Asn Ile Asn Ser Phe Asp Leu Lys Tyr 405 410 415 Ile Leu Thr Arg Leu Glu Tyr Leu Tyr Lys Val Asp Ser Gln Arg Phe 420 425 430 Cys Lys Leu Pro Thr Ala Gln Gly Gly Arg Phe Phe Leu His Ser Pro 435 440 445 Ala Val Gly Phe Lys Arg Gln Tyr Ala Ala Ala Phe Pro Ser Ala Ser 450 455 460 His Asn Asn Pro Ala Ser Thr Ala Ala Thr Lys Val Tyr Ile Ala Gly 465 470 475 480 Ser Val Val Ile Asp Met Tyr Pro Val Cys Met Ala Lys Thr Asn Ser 485 490 495 Pro Asn Tyr Lys Leu Asn Thr Met Ala Glu Leu Tyr Leu Arg Gln Arg 500 505 510 Lys Asp Asp Leu Ser Tyr Lys Asp Ile Pro Arg Cys Phe Val Ala Asn 515 520 525 Ala Glu Gly Arg Ala Gln Val Gly Arg Tyr Cys Leu Gln Asp Ala Val 530 535 540 Leu Val Arg Asp Leu Phe Asn Thr Ile Asn Phe His Tyr Glu Ala Gly 545 550 555 560 Ala Ile Ala Arg Leu Ala Lys Ile Pro Leu Arg Arg Val Ile Phe Asp 565 570 575 Gly Gln Gln Ile Arg Ile Tyr Thr Ser Leu Leu Asp Glu Cys Ala Cys 580 585 590 Arg Asp Phe Ile Leu Pro Asn His Tyr Ser Lys Gly Thr Thr Val Pro 595 600 605 Glu Thr Asn Ser Val Ala Val Ser Pro Asn Ala Ala Ile Ile Ser Thr 610 615 620 Ala Ala Val Pro Gly Asp Ala Gly Ser Val Ala Ala Met Phe Gln Met 625 630 635 640 Ser Pro Pro Leu Gln Ser Ala Pro Ser Ser Gln Asp Gly Val Ser Pro 645 650 655 Gly Ser Gly Ser Asn Ser Ser Ser Ser Val Gly Val Phe Ser Val Gly 660 665 670 Ser Gly Ser Ser Gly Gly Val Gly Val Ser Asn Asp Asn His Gly Ala 675 680 685 Gly Gly Thr Ala Ala Val Ser Tyr Gln Gly Ala Thr Val Phe Glu Pro 690 695 700 Glu Val Gly Tyr Tyr Asn Asp Pro Val Ala Val Phe Asp Phe Ala Ser 705 710 715 720 Leu Tyr Pro Ser Ile Ile Met Ala His Asn Leu Cys Tyr Ser Thr Leu 725 730 735 Leu Val Pro Gly Gly Glu Tyr Pro Val Asp Pro Ala Asp Val Tyr Ser 740 745 750 Val Thr Leu Glu Asn Gly Val Thr His Arg Phe Val Arg Ala Ser Val 755 760 765 Arg Val Ser Val Leu Ser Glu Leu Leu Asn Lys Trp Val Ser Gln Arg 770 775 780 Arg Ala Val Arg Glu Cys Met Arg Glu Cys Gln Asp Pro Val Arg Arg 785 790 795 800 Met Leu Leu Asp Lys Glu Gln Met Ala Leu Lys Val Thr Cys Asn Ala 805 810 815 Phe Tyr Gly Phe Thr Gly Val Val Asn Gly Met Met Pro Cys Leu Pro 820 825 830 Ile Ala Ala Ser Ile Thr Arg Ile Gly Arg Asp Met Leu Glu Arg Thr 835 840 845 Ala Arg Phe Ile Lys Asp Asn Phe Ser Glu Pro Cys Phe Leu His Asn 850 855 860 Phe Phe Asn Gln Glu Asp Tyr Val Val Gly Thr Arg Glu Gly Asp Ser 865 870 875 880 Glu Glu Ser Ser Ala Leu Pro Glu Gly Leu Glu Thr Ser Ser Gly Gly 885 890 895 Ser Asn Glu Arg Arg Val Glu Ala Arg Val Ile Tyr Gly Asp Thr Asp 900 905 910 Ser Val Phe Val Arg Phe Arg Gly Leu Thr Pro Gln Ala Leu Val Ala 915 920 925 Arg Gly Pro Ser Leu Ala His Tyr Val Thr Ala Cys Leu Phe Val Glu 930 935 940 Pro Val Lys Leu Glu Phe Glu Lys Val Phe Val Ser Leu Met Met Ile 945 950 955 960 Cys Lys Lys Arg Tyr Ile Gly Lys Val Glu Gly Ala Ser Gly Leu Ser 965 970 975 Met Lys Gly Val Asp Leu Val Arg Lys Thr Ala Cys Glu Phe Val Lys 980 985 990 Gly Val Thr Arg Asp Val Leu Ser Leu Leu Phe Glu Asp Arg Glu Val 995 1000 1005 Ser Glu Ala Ala Val Arg Leu Ser Arg Leu Ser Leu Asp Glu Val 1010 1015 1020 Lys Lys Tyr Gly Val Pro Arg Gly Phe Trp Arg Ile Leu Arg Arg 1025 1030 1035 Leu Val Gln Ala Arg Asp Asp Leu Tyr Leu His Arg Val Arg Val 1040 1045 1050 Glu Asp Leu Val Leu Ser Ser Val Leu Ser Lys Asp Ile Ser Leu 1055 1060 1065 Tyr Arg Gln Ser Asn Leu Pro His Ile Ala Val Ile Lys Arg Leu 1070 1075 1080 Ala Ala Arg Ser Glu Glu Leu Pro Ser Val Gly Asp Arg Val Phe 1085 1090 1095 Tyr Val Leu Thr Ala Pro Gly Val Arg Thr Ala Pro Gln Gly Ser 1100 1105 1110 Ser Asp Asn Gly Asp Ser Val Thr Ala Gly Val Val Ser Arg Ser 1115 1120 1125 Asp Ala Ile Asp Gly Thr Asp Asp Asp Ala Asp Gly Gly Gly Val 1130 1135 1140 Glu Glu Ser Asn Arg Arg Gly Gly Glu Pro Ala Lys Lys Arg Ala 1145 1150 1155 Arg Lys Pro Pro Ser Ala Val Cys Asn Tyr Glu Val Ala Glu Asp 1160 1165 1170 Pro Ser Tyr Val Arg Glu His Gly Val Pro Ile His Ala Asp Lys 1175 1180 1185 Tyr Phe Glu Gln Val Leu Lys Ala Val Thr Asn Val Leu Ser Pro 1190 1195 1200 Val Phe Pro Gly Gly Glu Thr Ala Arg Lys Asp Lys Phe Leu His 1205 1210 1215 Met Val Leu Pro Arg Arg Leu His Leu Glu Pro Ala Phe Leu Pro 1220 1225 1230 Tyr Ser Val Lys Ala His Glu Cys Cys 1235 1240 14 1238 PRT herpes simplex 14 Met Phe Cys Ala Ala Gly Gly Pro Thr Ser Pro Gly Gly Lys Ser Ala 1 5 10 15 Ala Arg Ala Ala Ser Gly Phe Phe Ala Pro His Asn Pro Arg Gly Ala 20 25 30 Thr Gln Thr Ala Pro Pro Pro Cys Arg Arg Gln Asn Phe Tyr Asn Pro 35 40 45 His Leu Ala Gln Thr Gly Thr Gln Pro Lys Ala Pro Gly Pro Ala Gln 50 55 60 Arg His Thr Tyr Tyr Ser Glu Cys Asp Glu Phe Arg Phe Ile Ala Pro 65 70 75 80 Arg Ser Leu Asp Glu Asp Ala Pro Ala Glu Gln Arg Thr Gly Val His 85 90 95 Asp Gly Arg Leu Arg Arg Ala Pro Lys Val Tyr Cys Gly Gly Asp Glu 100 105 110 Arg Asp Val Leu Arg Val Gly Pro Glu Gly Phe Trp Pro Arg Arg Leu 115 120 125 Arg Leu Trp Gly Gly Ala Asp His Ala Pro Lys Gly Phe Asp Pro Thr 130 135 140 Val Thr Val Phe His Val Tyr Asp Ile Leu Glu His Val Glu His Ala 145 150 155 160 Tyr Ser Met Arg Ala Ala Gln Leu His Glu Arg Phe Met Asp Ala Ile 165 170 175 Thr Pro Ala Gly Thr Val Ile Thr Leu Leu Gly Leu Thr Pro Glu Gly 180 185 190 His Arg Val Ala Val His Val Tyr Gly Thr Arg Gln Tyr Phe Tyr Met 195 200 205 Asn Lys Ala Glu Val Asp Arg His Leu Gln Cys Arg Ala Pro Arg Asp 210 215 220 Leu Cys Glu Arg Leu Ala Ala Ala Leu Arg Glu Ser Pro Gly Ala Ser 225 230 235 240 Phe Arg Gly Ile Ser Ala Asp His Phe Glu Ala Glu Val Val Glu Arg 245 250 255 Ala Asp Val Tyr Tyr Tyr Glu Thr Arg Pro Thr Leu Tyr Tyr Arg Val 260 265 270 Phe Val Arg Ser Gly Arg Ala Leu Ala Tyr Leu Cys Asp Asn Phe Cys 275 280 285 Pro Ala Ile Arg Lys Tyr Glu Gly Gly Val Asp Ala Thr Thr Arg Phe 290 295 300 Ile Leu Asp Asn Pro Gly Phe Val Thr Phe Gly Trp Tyr Arg Leu Lys 305 310 315 320 Pro Gly Arg Gly Asn Ala Pro Ala Gln Pro Arg Pro Pro Thr Ala Phe 325 330 335 Gly Thr Ser Ser Asp Val Glu Phe Asn Cys Thr Ala Asp Asn Leu Ala 340 345 350 Val Glu Gly Ala Met Cys Asp Leu Pro Ala Tyr Lys Leu Met Cys Phe 355 360 365 Asp Ile Glu Cys Lys Ala Gly Gly Glu Asp Glu Leu Ala Phe Pro Val 370 375 380 Ala Glu Arg Pro Glu Asp Leu Val Ile Gln Ile Ser Cys Leu Leu Tyr 385 390 395 400 Asp Leu Ser Thr Thr Ala Leu Glu His Ile Leu Leu Phe Ser Leu Gly 405 410 415 Ser Cys Asp Leu Pro Glu Ser His Leu Ser Asp Leu Ala Ser Arg Gly 420 425 430 Leu Pro Ala Pro Val Val Leu Glu Phe Asp Ser Glu Phe Glu Met Leu 435 440 445 Leu Ala Phe Met Thr Phe Val Lys Gln Tyr Gly Pro Glu Phe Val Thr 450 455 460 Gly Tyr Asn Ile Ile Asn Phe Asp Trp Pro Phe Val Leu Thr Lys Leu 465 470 475 480 Thr Glu Ile Tyr Lys Val Pro Leu Asp Gly Tyr Gly Arg Met Asn Gly 485 490 495 Arg Gly Val Phe Arg Val Trp Asp Ile Gly Gln Ser His Phe Gln Lys 500 505 510 Arg Ser Lys Ile Lys Val Asn Gly Met Val Asn Ile Asp Met Tyr Gly 515 520 525 Ile Ile Thr Asp Lys Val Lys Leu Ser Ser Tyr Lys Leu Asn Ala Val 530 535 540 Ala Glu Ala Val Leu Lys Asp Lys Lys Lys Asp Leu Ser Tyr Arg Asp 545 550 555 560 Ile Pro Ala Tyr Tyr Ala Ser Gly Pro Ala Gln Arg Gly Val Ile Gly 565 570 575 Glu Tyr Cys Val Gln Asp Ser Leu Leu Val Gly Gln Leu Phe Phe Lys 580 585 590 Phe Leu Pro His Leu Glu Leu Ser Ala Val Ala Arg Leu Ala Gly Ile 595 600 605 Asn Ile Thr Arg Thr Ile Tyr Asp Gly Gln Gln Ile Arg Val Phe Thr 610 615 620 Cys Leu Leu Arg Leu Ala Gly Gln Lys Gly Phe Ile Leu Pro Asp Thr 625 630 635 640 Gln Gly Arg Phe Arg Gly Leu Asp Lys Glu Ala Pro Lys Arg Pro Ala 645 650 655 Val Pro Arg Gly Glu Gly Glu Arg Pro Gly Asp Gly Asn Gly Asp Glu 660 665 670 Asp Lys Asp Asp Asp Glu Asp Glu Asp Gly Asp Glu Arg Glu Glu Val 675 680 685 Ala Arg Glu Thr Gly Gly Arg His Val Gly Tyr Gln Gly Ala Arg Val 690 695 700 Leu Asp Pro Thr Ser Gly Phe His Val Asp Pro Val Val Val Phe Asp 705 710 715 720 Phe Ala Ser Leu Tyr Pro Ser Ile Ile Gln Ala His Asn Leu Cys Phe 725 730 735 Ser Thr Leu Ser Leu Arg Pro Glu Ala Val Ala His Leu Glu Ala Asp 740 745 750 Arg Asp Tyr Leu Glu Ile Glu Val Gly Gly Arg Arg Leu Phe Phe Val 755 760 765 Lys Ala His Val Arg Glu Ser Leu Leu Ser Ile Leu Leu Arg Asp Trp 770 775 780 Leu Ala Met Arg Lys Gln Ile Arg Ser Arg Ile Pro Gln Ser Thr Pro 785 790 795 800 Glu Glu Ala Val Leu Leu Asp Lys Gln Gln Ala Ala Ile Lys Val Val 805 810 815 Cys Asn Ser Val Tyr Gly Phe Thr Gly Val Gln His Gly Leu Leu Pro 820 825 830 Cys Leu His Val Ala Ala Thr Val Thr Thr Ile Gly Arg Glu Met Leu 835 840 845 Leu Ala Thr Arg Ala Tyr Val His Ala Arg Trp Ala Glu Phe Asp Gln 850 855 860 Leu Leu Ala Asp Phe Pro Glu Ala Ala Gly Met Arg Ala Pro Gly Pro 865 870 875 880 Tyr Ser Met Arg Ile Ile Tyr Gly Asp Thr Asp Ser Ile Phe Val Leu 885 890 895 Cys Arg Gly Leu Thr Ala Ala Gly Leu Val Ala Met Gly Asp Lys Met 900 905 910 Ala Ser His Ile Ser Arg Ala Leu Phe Leu Pro Pro Ile Lys Leu Glu 915 920 925 Cys Glu Lys Thr Phe Thr Lys Leu Leu Leu Ile Ala Lys Lys Lys Tyr 930 935 940 Ile Gly Val Ile Cys Gly Gly Lys Met Leu Ile Lys Gly Val Asp Leu 945 950 955 960 Val Arg Lys Asn Asn Cys Ala Phe Ile Asn Arg Thr Ser Arg Ala Leu 965 970 975 Val Asp Leu Leu Phe Tyr Asp Asp Thr Val Ser Gly Ala Ala Ala Ala 980 985 990 Leu Ala Glu Arg Pro Ala Glu Glu Trp Leu Ala Arg Pro Leu Pro Glu 995 1000 1005 Gly Leu Gln Ala Phe Gly Ala Val Leu Val Asp Ala His Arg Arg 1010 1015 1020 Ile Thr Asp Pro Glu Arg Asp Ile Gln Asp Phe Val Leu Thr Ala 1025 1030 1035 Glu Leu Ser Arg His Pro Arg Ala Tyr Thr Asn Lys Arg Leu Ala 1040 1045 1050 His Leu Thr Val Tyr Tyr Lys Leu Met Ala Arg Arg Ala Gln Val 1055 1060 1065 Pro Ser Ile Lys Asp Arg Ile Pro Tyr Val Ile Val Ala Gln Thr 1070 1075 1080 Arg Glu Val Glu Glu Thr Val Ala Arg Leu Ala Ala Leu Arg Glu 1085 1090 1095 Leu Asp Ala Ala Ala Pro Gly Asp Glu Pro Ala Pro Pro Ala Ala 1100 1105 1110 Leu Pro Ser Pro Ala Lys Arg Pro Arg Glu Thr Pro Ser His Ala 1115 1120 1125 Asp Pro Pro Gly Gly Ala Ser Lys Pro Arg Lys Leu Leu Val Ser 1130 1135 1140 Glu Leu Ala Glu Asp Pro Gly Tyr Ala Ile Ala Arg Gly Val Pro 1145 1150 1155 Leu Asn Thr Asp Tyr Tyr Phe Ser His Leu Leu Gly Ala Ala Cys 1160 1165 1170 Val Thr Phe Lys Ala Leu Phe Gly Asn Asn Ala Lys Ile Thr Glu 1175 1180 1185 Ser Leu Leu Lys Arg Phe Ile Pro Glu Thr Trp His Pro Pro Asp 1190 1195 1200 Asp Val Ala Ala Arg Leu Arg Ala Ala Gly Phe Gly Pro Ala Gly 1205 1210 1215 Ala Gly Ala Thr Ala Glu Glu Thr Arg Arg Met Leu His Arg Ala 1220 1225 1230 Phe Asp Thr Leu Ala 1235 15 1240 PRT herpes simplex 15 Met Phe Cys Ala Ala Gly Gly Pro Ala Ser Pro Gly Gly Lys Ser Ala 1 5 10 15 Ala Arg Ala Ala Ser Gly Phe Phe Ala Pro His Asn Pro Arg Gly Ala 20 25 30 Thr Gln Thr Ala Pro Pro Pro Cys Arg Arg Gln Asn Phe Tyr Asn Pro 35 40 45 His Leu Ala Gln Thr Gly Thr Gln Pro Lys Ala Pro Gly Pro Ala Gln 50 55 60 Arg His Thr Tyr Tyr Ser Glu Cys Asp Glu Phe Arg Phe Ile Ala Pro 65 70 75 80 Arg Ser Leu Asp Glu Asp Ala Pro Ala Glu Gln Arg Thr Gly Val His 85 90 95 Asp Gly Arg Leu Arg Arg Ala Pro Lys Val Tyr Cys Gly Gly Asp Glu 100 105 110 Arg Asp Val Leu Arg Val Gly Pro Glu Gly Phe Trp Pro Arg Arg Leu 115 120 125 Arg Leu Trp Gly Gly Ala Asp His Ala Pro Glu Gly Phe Asp Pro Thr 130 135 140 Val Thr Val Phe His Val Tyr Asp Ile Leu Glu His Val Glu His Ala 145 150 155 160 Tyr Ser Met Arg Ala Ala Gln Leu His Glu Arg Phe Met Asp Ala Ile 165 170 175 Thr Pro Ala Gly Thr Val Ile Thr Leu Leu Gly Leu Thr Pro Glu Gly 180 185 190 His Arg Val Ala Val His Val Tyr Gly Thr Arg Gln Tyr Phe Tyr Met 195 200 205 Asn Lys Ala Glu Val Asp Arg His Leu Gln Cys Arg Ala Pro Arg Asp 210 215 220 Leu Cys Glu Arg Leu Ala Ala Ala Leu Arg Glu Ser Pro Gly Ala Ser 225 230 235 240 Phe Arg Gly Ile Ser Ala Asp His Phe Glu Ala Glu Val Val Glu Arg 245 250 255 Ala Asp Val Tyr Tyr Tyr Glu Thr Arg Pro Thr Leu Tyr Tyr Arg Val 260 265 270 Phe Val Arg Ser Gly Arg Ala Leu Ala Tyr Leu Cys Asp Asn Phe Cys 275 280 285 Pro Ala Ile Arg Lys Tyr Glu Gly Gly Val Asp Ala Thr Thr Arg Phe 290 295 300 Ile Leu Asp Asn Pro Gly Phe Val Thr Phe Gly Trp Tyr Arg Leu Lys 305 310 315 320 Pro Gly Arg Gly Asn Ala Pro Ala Gln Pro Arg Pro Pro Thr Ala Phe 325 330 335 Gly Thr Ser Ser Asp Val Glu Phe Asn Cys Thr Ala Asp Asn Leu Ala 340 345 350 Val Glu Gly Ala Met Cys Asp Leu Pro Ala Tyr Lys Leu Met Cys Phe 355 360 365 Asp Ile Glu Cys Lys Ala Gly Gly Glu Asp Glu Leu Ala Phe Pro Val 370 375 380 Ala Glu Arg Pro Glu Asp Leu Val Ile Gln Ile Ser Cys Leu Leu Tyr 385 390 395 400 Asp Leu Ser Thr Thr Ala Leu Glu His Ile Leu Leu Phe Ser Leu Gly 405 410 415 Ser Cys Asp Leu Pro Glu Ser His Leu Ser Asp Leu Ala Ser Arg Gly 420 425 430 Leu Pro Ala Pro Val Val Leu Glu Phe Asp Ser Glu Phe Glu Met Leu 435 440 445 Leu Ala Phe Met Thr Phe Val Lys Gln Tyr Gly Pro Glu Phe Val Thr 450 455 460 Gly Tyr Asn Ile Ile Asn Phe Asp Trp Pro Phe Val Leu Thr Lys Leu 465 470 475 480 Thr Glu Ile Tyr Lys Val Pro Leu Asp Gly Tyr Gly Arg Met Asn Gly 485 490 495 Arg Gly Val Phe Arg Val Trp Asp Ile Gly Gln Ser His Phe Gln Lys 500 505 510 Arg Ser Lys Ile Lys Val Asn Gly Met Val Asn Ile Asp Met Tyr Gly 515 520 525 Ile Ile Thr Asp Lys Val Lys Leu Ser Ser Tyr Lys Leu Asn Ala Val 530 535 540 Ala Glu Ala Val Leu Lys Asp Lys Lys Lys Asp Leu Ser Tyr Arg Asp 545 550 555 560 Ile Pro Ala Tyr Tyr Ala Ser Gly Pro Ala Gln Arg Gly Val Ile Gly 565 570 575 Glu Tyr Cys Val Gln Asp Ser Leu Leu Val Gly Gln Leu Phe Phe Lys 580 585 590 Phe Leu Pro His Leu Glu Leu Ser Ala Val Ala Arg Leu Ala Gly Ile 595 600 605 Asn Ile Thr Arg Thr Ile Tyr Asp Gly Gln Gln Ile Arg Val Phe Thr 610 615 620 Cys Leu Leu Arg Leu Ala Gly Gln Lys Gly Phe Ile Leu Pro Asp Thr 625 630 635 640 Gln Gly Arg Phe Arg Gly Leu Asp Lys Glu Ala Pro Lys Arg Pro Ala 645 650 655 Val Pro Arg Gly Glu Gly Glu Arg Pro Gly Asp Gly Asn Gly Asp Glu 660 665 670 Asp Lys Asp Asp Asp Glu Asp Gly Asp Glu Asp Gly Asp Glu Arg Glu 675 680 685 Glu Val Ala Arg Glu Thr Gly Gly Arg His Val Gly Tyr Gln Gly Ala 690 695 700 Arg Val Leu Asp Pro Thr Ser Gly Phe His Val Asp Pro Val Val Val 705 710 715 720 Phe Asp Phe Ala Ser Leu Tyr Pro Ser Ile Ile Gln Ala His Asn Leu 725 730 735 Cys Phe Ser Thr Leu Ser Leu Arg Pro Glu Ala Val Ala His Leu Glu 740 745 750 Ala Asp Arg Asp Tyr Leu Glu Ile Glu Val Gly Gly Arg Arg Leu Phe 755 760 765 Phe Val Lys Ala His Val Arg Glu Ser Leu Leu Ser Ile Leu Leu Arg 770 775 780 Asp Trp Leu Ala Met Arg Lys Gln Ile Arg Ser Arg Ile Pro Gln Ser 785 790 795 800 Pro Pro Glu Glu Ala Val Leu Leu Asp Lys Gln Gln Ala Ala Ile Lys 805 810 815 Val Val Cys Asn Ser Val Tyr Gly Phe Thr Gly Val Gln His Gly Leu 820 825 830 Leu Pro Cys Leu His Val Ala Ala Thr Val Thr Thr Ile Gly Arg Glu 835 840 845 Met Leu Leu Ala Thr Arg Ala Tyr Val His Ala Arg Trp Ala Glu Phe 850 855 860 Asp Gln Leu Leu Ala Asp Phe Pro Glu Ala Ala Gly Met Arg Ala Pro 865 870 875 880 Gly Pro Tyr Ser Met Arg Ile Ile Tyr Gly Asp Thr Asp Ser Ile Phe 885 890 895 Val Leu Cys Arg Gly Leu Thr Ala Ala Gly Leu Val Ala Met Gly Asp 900 905 910 Lys Met Ala Ser His Ile Ser Arg Ala Leu Phe Leu Pro Pro Ile Lys 915 920 925 Leu Glu Cys Glu Lys Thr Phe Thr Lys Leu Leu Leu Ile Ala Lys Lys 930 935 940 Lys Tyr Ile Gly Val Ile Cys Gly Gly Lys Met Leu Ile Lys Gly Val 945 950 955 960 Asp Leu Val Arg Lys Asn Asn Cys Ala Phe Ile Asn Arg Thr Ser Arg 965 970 975 Ala Leu Val Asp Leu Leu Phe Tyr Asp Asp Thr Val Ser Gly Ala Ala 980 985 990 Ala Ala Leu Ala Glu Arg Pro Ala Glu Glu Trp Leu Ala Arg Pro Leu 995 1000 1005 Pro Glu Gly Leu Gln Ala Phe Gly Ala Val Leu Val Asp Ala His 1010 1015 1020 Arg Arg Ile Thr Asp Pro Glu Arg Asp Ile Gln Asp Phe Val Leu 1025 1030 1035 Thr Ala Glu Leu Ser Arg His Pro Arg Ala Tyr Thr Asn Lys Arg 1040 1045 1050 Leu Ala His Leu Thr Val Tyr Tyr Lys Leu Met Ala Arg Arg Ala 1055 1060 1065 Gln Val Pro Ser Ile Lys Asp Arg Ile Pro Tyr Val Ile Val Ala 1070 1075 1080 Gln Thr Arg Glu Val Glu Glu Thr Val Ala Arg Leu Ala Ala Leu 1085 1090 1095 Arg Glu Leu Asp Ala Ala Ala Pro Gly Asp Glu Pro Ala Pro Pro 1100 1105 1110 Ala Ala Leu Pro Ser Pro Ala Lys Arg Pro Arg Glu Thr Pro Ser 1115 1120 1125 His Ala Asp Pro Pro Gly Gly Ala Ser Lys Pro Arg Lys Leu Leu 1130 1135 1140 Val Ser Glu Leu Ala Glu Asp Pro Gly Tyr Ala Ile Ala Arg Gly 1145 1150 1155 Val Pro Leu Asn Thr Asp Tyr Tyr Phe Ser His Leu Leu Gly Ala 1160 1165 1170 Ala Cys Val Thr Phe Lys Ala Leu Phe Gly Asn Asn Ala Lys Ile 1175 1180 1185 Thr Glu Ser Leu Leu Lys Arg Phe Ile Pro Glu Thr Trp His Pro 1190 1195 1200 Pro Asp Asp Val Ala Ala Arg Leu Arg Ala Ala Gly Phe Gly Pro 1205 1210 1215 Ala Gly Ala Gly Ala Thr Ala Glu Glu Thr Arg Arg Met Leu His 1220 1225 1230 Arg Ala Phe Asp Thr Leu Ala 1235 1240 16 1235 PRT herpes simplex 16 Met Phe Ser Gly Gly Gly Gly Pro Leu Ser Pro Gly Gly Lys Ser Ala 1 5 10 15 Ala Arg Ala Ala Ser Gly Phe Phe Ala Pro Ala Gly Pro Arg Gly Ala 20 25 30 Gly Arg Gly Pro Pro Pro Cys Leu Arg Gln Asn Phe Tyr Asn Pro Tyr 35 40 45 Leu Ala Pro Val Gly Thr Gln Gln Lys Pro Thr Gly Pro Thr Gln Arg 50 55 60 His Thr Tyr Tyr Ser Glu Cys Asp Glu Phe Arg Phe Ile Ala Pro Arg 65 70 75 80 Val Leu Asp Glu Asp Ala Pro Pro Glu Lys Arg Ala Gly Val His Asp 85 90 95 Gly His Leu Lys Arg Ala Pro Lys Val Tyr Cys Gly Gly Asp Glu Arg 100 105 110 Asp Val Leu Arg Val Gly Ser Gly Gly Phe Trp Pro Arg Arg Ser Arg 115 120 125 Leu Trp Gly Gly Val Asp His Ala Pro Ala Gly Phe Asn Pro Thr Val 130 135 140 Thr Val Phe His Val Tyr Asp Ile Leu Glu Asn Val Glu His Ala Tyr 145 150 155 160 Gly Met Arg Ala Ala Gln Phe His Ala Arg Phe Met Asp Ala Ile Thr 165 170 175 Pro Thr Gly Thr Val Ile Thr Leu Leu Gly Leu Thr Pro Glu Gly His 180 185 190 Arg Val Ala Val His Val Tyr Gly Thr Arg Gln Tyr Phe Tyr Met Asn 195 200 205 Lys Glu Glu Val Asp Arg His Leu Gln Cys Arg Ala Pro Arg Asp Leu 210 215 220 Cys Glu Arg Met Ala Ala Ala Leu Arg Glu Ser Pro Gly Ala Ser Phe 225 230 235 240 Arg Gly Ile Ser Ala Asp His Phe Glu Ala Glu Val Val Glu Arg Thr 245 250 255 Asp Val Tyr Tyr Tyr Glu Thr Arg Pro Ala Leu Phe Tyr Arg Val Tyr 260 265 270 Val Arg Ser Gly Arg Val Leu Ser Tyr Leu Cys Asp Asn Phe Cys Pro 275 280 285 Ala Ile Lys Lys Tyr Glu Gly Gly Val Asp Ala Thr Thr Arg Phe Ile 290 295 300 Leu Asp Asn Pro Gly Phe Val Thr Phe Gly Trp Tyr Arg Leu Lys Pro 305 310 315 320 Gly Arg Asn Asn Thr Leu Ala Gln Pro Arg Ala Pro Met Ala Phe Gly 325 330 335 Thr Ser Ser Asp Val Glu Phe Asn Cys Thr Ala Asp Asn Leu Ala Ile 340 345 350 Glu Gly Gly Met Ser Asp Leu Pro Ala Tyr Lys Leu Met Cys Phe Asp 355 360 365 Ile Glu Cys Lys Ala Gly Gly Glu Asp Glu Leu Ala Phe Pro Val Ala 370 375 380 Gly His Pro Glu Asp Leu Val Ile Gln Ile Ser Cys Leu Leu Tyr Asp 385 390 395 400 Leu Ser Thr Thr Ala Leu Glu His Val Leu Leu Phe Ser Leu Gly Ser 405 410 415 Cys Asp Leu Pro Glu Ser His Leu Asn Glu Leu Ala Ala Arg Gly Leu 420 425 430 Pro Thr Pro Val Val Leu Glu Phe Asp Ser Glu Phe Glu Met Leu Leu 435 440 445 Ala Phe Met Thr Leu Val Lys Gln Tyr Gly Pro Glu Phe Val Thr Gly 450 455 460 Tyr Asn Ile Ile Asn Phe Asp Trp Pro Phe Leu Leu Ala Lys Leu Thr 465 470 475 480 Asp Ile Tyr Lys Val Pro Leu Asp Gly Tyr Gly Arg Met Asn Gly Arg 485 490 495 Gly Val Phe Arg Val Trp Asp Ile Gly Gln Ser His Phe Gln Lys Arg 500 505 510 Ser Lys Ile Lys Val Asn Gly Met Val Asn Ile Asp Met Tyr Gly Ile 515 520 525 Ile Thr Asp Lys Ile Lys Leu Ser Ser Tyr Lys Leu Asn Ala Val Ala 530 535 540 Glu Ala Val Leu Lys Asp Lys Lys Lys Asp Leu Ser Tyr Arg Asp Ile 545 550 555 560 Pro Ala Tyr Tyr Ala Ala Gly Pro Ala Gln Arg Gly Val Ile Gly Glu 565 570 575 Tyr Cys Ile Gln Asp Ser Leu Leu Val Gly Gln Leu Phe Phe Lys Phe 580 585 590 Leu Pro His Leu Glu Leu Ser Ala Val Ala Arg Leu Ala Gly Ile Asn 595 600 605 Ile Thr Arg Thr Ile Tyr Asp Gly Gln Gln Ile Arg Val Phe Thr Cys 610 615 620 Leu Leu Arg Leu Ala Asp Gln Lys Gly Phe Ile Leu Pro Asp Thr Gln 625 630 635 640 Gly Arg Phe Arg Gly Ala Gly Gly Glu Ala Pro Lys Arg Pro Ala Ala 645 650 655 Ala Arg Glu Asp Glu Glu Arg Pro Glu Glu Glu Gly Glu Asp Glu Asp 660 665 670 Glu Arg Glu Glu Gly Gly Gly Glu Arg Glu Pro Glu Gly Ala Arg Glu 675 680 685 Thr Ala Gly Arg His Val Gly Tyr Gln Gly Ala Arg Val Leu Asp Pro 690 695 700 Thr Ser Gly Phe His Val Asn Pro Val Val Val Phe Asp Phe Ala Ser 705 710 715 720 Leu Tyr Pro Ser Ile Ile Gln Ala His Asn Leu Cys Phe Ser Thr Leu 725 730 735 Ser Leu Arg Ala Asp Ala Val Ala His Leu Glu Ala Gly Lys Asp Tyr 740 745 750 Leu Glu Ile Glu Val Gly Gly Arg Arg Leu Phe Phe Val Lys Ala His 755 760 765 Val Arg Glu Ser Leu Leu Ser Ile Leu Leu Arg Asp Trp Leu Ala Met 770 775 780 Arg Lys Gln Ile Arg Ser Arg Ile Pro Gln Ser Ser Pro Glu Glu Ala 785 790 795 800 Val Leu Leu Asp Lys Gln Gln Ala Ala Ile Lys Val Val Cys Asn Ser 805 810 815 Val Tyr Gly Phe Thr Gly Val Gln His Gly Leu Leu Pro Cys Leu His 820 825 830 Val Ala Ala Thr Val Thr Thr Ile Gly Arg Glu Met Leu Leu Ala Thr 835 840 845 Arg Glu Tyr Val His Ala Arg Trp Ala Ala Phe Glu Gln Leu Leu Ala 850 855 860 Asp Phe Pro Glu Ala Ala Asp Met Arg Ala Pro Gly Pro Tyr Ser Met 865 870 875 880 Arg Ile Ile Tyr Gly Asp Thr Asp Ser Ile Phe Val Leu Cys Arg Gly 885 890 895 Leu Thr Ala Ala Gly Leu Thr Ala Met Gly Asp Lys Met Ala Ser His 900 905 910 Ile Ser Arg Ala Leu Phe Leu Pro Pro Ile Lys Leu Glu Cys Glu Lys 915 920 925 Thr Phe Thr Lys Leu Leu Leu Ile Ala Lys Lys Lys Tyr Ile Gly Val 930 935 940 Ile Tyr Gly Gly Lys Met Leu Ile Lys Gly Val Asp Leu Val Arg Lys 945 950 955 960 Asn Asn Cys Ala Phe Ile Asn Arg Thr Ser Arg Ala Leu Val Asp Leu 965 970 975 Leu Phe Tyr Asp Asp Thr Val Ser Gly Ala Ala Ala Ala Leu Ala Glu 980 985 990 Arg Pro Ala Glu Glu Trp Leu Ala Arg Pro Leu Pro Glu Gly Leu Gln 995 1000 1005 Ala Phe Gly Ala Val Leu Val Asp Ala His Arg Arg Ile Thr Asp 1010 1015 1020 Pro Glu Arg Asp Ile Gln Asp Phe Val Leu Thr Ala Glu Leu Ser 1025 1030 1035 Arg His Pro Arg Ala Tyr Thr Asn Lys Arg Leu Ala His Leu Thr 1040 1045 1050 Val Tyr Tyr Lys Leu Met Ala Arg Arg Ala Gln Val Pro Ser Ile 1055 1060 1065 Lys Asp Arg Ile Pro Tyr Val Ile Val Ala Gln Thr Arg Glu Val 1070 1075 1080 Glu Glu Thr Val Ala Arg Leu Ala Ala Leu Arg Glu Leu Asp Ala 1085 1090 1095 Ala Ala Pro Gly Asp Glu Pro Ala Pro Pro Ala Ala Leu Pro Ser 1100 1105 1110 Pro Ala Lys Arg Pro Arg Glu Thr Pro Ser His Ala Asp Pro Pro 1115 1120 1125 Gly Gly Ala Ser Lys Pro Arg Lys Leu Leu Val Ser Glu Leu Ala 1130 1135 1140 Glu Asp Pro Ala Tyr Ala Ile Ala His Gly Val Ala Leu Asn Thr 1145 1150 1155 Asp Tyr Tyr Phe Ser His Leu Leu Gly Ala Ala Cys Val Thr Phe 1160 1165 1170 Lys Ala Leu Phe Gly Asn Asn Ala Lys Ile Thr Glu Ser Leu Leu 1175 1180 1185 Lys Arg Phe Ile Pro Glu Val Trp His Pro Pro Asp Asp Val Ala 1190 1195 1200 Ala Arg Leu Arg Ala Ala Gly Phe Gly Ala Val Gly Ala Gly Ala 1205 1210 1215 Thr Ala Glu Glu Thr Arg Arg Met Leu His Arg Ala Phe Asp Thr 1220 1225 1230 Leu Ala 1235 17 1235 PRT herpes simplex 17 Met Phe Ser Gly Gly Gly Gly Pro Leu Ser Pro Gly Gly Lys Ser Ala 1 5 10 15 Ala Arg Ala Ala Ser Gly Phe Phe Ala Pro Ala Gly Pro Arg Gly Ala 20 25 30 Gly Arg Gly Pro Pro Pro Cys Leu Arg Gln Asn Phe Tyr Asn Pro Tyr 35 40 45 Leu Ala Pro Val Gly Thr Gln Gln Lys Pro Thr Gly Pro Thr Gln Arg 50 55 60 His Thr Tyr Tyr Ser Glu Cys Asp Glu Phe Arg Phe Ile Ala Pro Arg 65 70 75 80 Val Leu Asp Glu Asp Ala Pro Pro Glu Lys Arg Ala Gly Val His Asp 85 90 95 Gly His Leu Lys Arg Ala Pro Lys Val Tyr Cys Gly Gly Asp Glu Arg 100 105 110 Asp Val Leu Arg Val Gly Ser Gly Gly Phe Trp Pro Arg Arg Ser Arg 115 120 125 Leu Trp Gly Gly Val Asp His Ala Pro Ala Gly Phe Asn Pro Thr Val 130 135 140 Thr Val Phe His Val Tyr Asp Ile Leu Glu Asn Val Glu His Ala Tyr 145 150 155 160 Gly Met Arg Ala Ala Gln Phe His Ala Arg Phe Met Asp Ala Ile Thr 165 170 175 Pro Thr Gly Thr Val Ile Thr Leu Leu Gly Leu Thr Pro Glu Gly His 180 185 190 Arg Val Ala Val His Val Tyr Gly Thr Arg Gln Tyr Phe Tyr Met Asn 195 200 205 Lys Glu Glu Val Asp Arg His Leu Gln Cys Arg Ala Pro Arg Asp Leu 210 215 220 Cys Glu Arg Met Ala Ala Ala Leu Arg Glu Ser Pro Gly Ala Ser Phe 225 230 235 240 Arg Gly Ile Ser Ala Asp His Phe Glu Ala Glu Val Val Glu Arg Thr 245 250 255 Asp Val Tyr Tyr Tyr Glu Thr Arg Pro Ala Leu Phe Tyr Arg Val Tyr 260 265 270 Val Arg Ser Gly Arg Val Leu Ser Tyr Leu Cys Asp Asn Phe Cys Pro 275 280 285 Ala Ile Lys Lys Tyr Glu Gly Gly Val Asp Ala Thr Thr Arg Phe Ile 290 295 300 Leu Asp Asn Pro Gly Phe Val Thr Phe Gly Trp Tyr Arg Leu Lys Pro 305 310 315 320 Gly Arg Asn Asn Thr Leu Ala Gln Pro Arg Ala Pro Met Ala Phe Gly 325 330 335 Thr Ser Ser Asp Val Glu Phe Asn Cys Thr Ala Asp Asn Leu Ala Ile 340 345 350 Glu Gly Gly Met Ser Asp Leu Pro Ala Tyr Lys Leu Met Cys Phe Asp 355 360 365 Ile Glu Cys Lys Ala Gly Gly Glu Asp Glu Leu Ala Phe Pro Val Ala 370 375 380 Gly His Pro Glu Asp Leu Val Ile Gln Ile Ser Cys Leu Leu Tyr Asp 385 390 395 400 Leu Ser Thr Thr Ala Leu Glu His Val Leu Leu Phe Ser Leu Gly Ser 405 410 415 Cys Asp Leu Pro Glu Ser His Leu Asn Glu Leu Ala Ala Arg Gly Leu 420 425 430 Pro Thr Pro Val Val Leu Glu Phe Asp Ser Glu Phe Glu Met Leu Leu 435 440 445 Ala Phe Met Thr Leu Val Lys Gln Tyr Gly Pro Glu Phe Val Thr Gly 450 455 460 Tyr Asn Ile Ile Asn Phe Asp Trp Pro Phe Leu Leu Ala Lys Leu Thr 465 470 475 480 Asp Ile Tyr Lys Val Pro Leu Asp Gly Tyr Gly Arg Met Asn Gly Arg 485 490 495 Gly Val Phe Arg Val Trp Asp Ile Gly Gln Ser His Phe Gln Lys Arg 500 505 510 Ser Lys Ile Lys Val Asn Gly Met Val Asn Ile Asp Met Tyr Gly Ile 515 520 525 Ile Thr Asp Lys Ile Lys Leu Ser Ser Tyr Lys Leu Asn Ala Val Ala 530 535 540 Glu Ala Val Leu Lys Asp Lys Lys Lys Asp Leu Ser Tyr Arg Asp Ile 545 550 555 560 Pro Ala Tyr Tyr Ala Ala Gly Pro Ala Gln Arg Gly Val Ile Gly Glu 565 570 575 Tyr Cys Ile Gln Asp Ser Leu Leu Val Gly Gln Leu Phe Phe Lys Phe 580 585 590 Leu Pro His Leu Glu Leu Ser Ala Val Ala Arg Leu Ala Gly Ile Asn 595 600 605 Ile Thr Arg Thr Ile Tyr Asp Gly Gln Gln Ile Arg Val Phe Thr Cys 610 615 620 Leu Leu Arg Leu Ala Asp Gln Lys Gly Phe Ile Leu Pro Asp Thr Gln 625 630 635 640 Gly Arg Phe Arg Gly Ala Gly Gly Glu Ala Pro Lys Arg Pro Ala Ala 645 650 655 Ala Arg Glu Asp Glu Glu Arg Pro Glu Glu Glu Gly Glu Asp Glu Asp 660 665 670 Glu Arg Glu Glu Gly Gly Gly Glu Arg Glu Pro Glu Gly Ala Arg Glu 675 680 685 Thr Ala Gly Arg His Val Gly Tyr Gln Gly Ala Arg Val Leu Asp Pro 690 695 700 Ile Ser Gly Phe His Val Asn Pro Val Val Val Phe Asp Phe Ala Ser 705 710 715 720 Leu Tyr Pro Ser Ile Ile Gln Ala His Asn Leu Cys Phe Ser Thr Leu 725 730 735 Ser Leu Arg Ala Asp Ala Val Ala His Leu Glu Ala Gly Lys Asp Tyr 740 745 750 Leu Glu Ile Glu Val Gly Gly Arg Arg Leu Phe Phe Val Lys Ala His 755 760 765 Val Arg Glu Ser Leu Leu Ser Ile Leu Leu Arg Asp Trp Leu Ala Met 770 775 780 Arg Lys Gln Ile Arg Ser Arg Ile Pro Gln Ser Ser Pro Glu Glu Ala 785 790 795 800 Val Leu Leu Asp Lys Gln Gln Ala Ala Ile Lys Val Val Cys Asn Ser 805 810 815 Val Tyr Gly Phe Thr Gly Val Gln His Gly Leu Leu Pro Cys Leu His 820 825 830 Val Ala Ala Thr Val Thr Thr Ile Gly Arg Glu Met Leu Leu Ala Thr 835 840 845 Arg Glu Tyr Val His Ala Arg Trp Ala Ala Phe Glu Gln Leu Leu Ala 850 855 860 Asp Phe Pro Glu Ala Ala Asp Met Arg Ala Pro Gly Pro Tyr Ser Met 865 870 875 880 Arg Ile Ile Tyr Gly Asp Thr Asp Ser Ile Phe Val Leu Cys Arg Gly 885 890 895 Leu Thr Ala Ala Gly Leu Thr Ala Met Gly Asp Lys Met Ala Ser His 900 905 910 Ile Ser Arg Ala Leu Phe Leu Pro Pro Ile Lys Leu Glu Cys Glu Lys 915 920 925 Thr Phe Thr Lys Leu Leu Leu Ile Ala Lys Lys Lys Tyr Ile Gly Val 930 935 940 Ile Tyr Gly Gly Lys Met Leu Ile Lys Gly Val Asp Leu Val Arg Lys 945 950 955 960 Asn Asn Cys Ala Phe Ile Asn Arg Thr Ser Arg Ala Leu Val Asp Leu 965 970 975 Leu Phe Tyr Asp Asp Thr Val Ser Gly Ala Ala Ala Ala Leu Ala Glu 980 985 990 Arg Pro Ala Glu Glu Trp Leu Ala Arg Pro Leu Pro Glu Gly Leu Gln 995 1000 1005 Ala Phe Gly Ala Val Leu Val Asp Ala His Arg Arg Ile Thr Asp 1010 1015 1020 Pro Glu Arg Asp Ile Gln Asp Phe Val Leu Thr Ala Glu Leu Ser 1025 1030 1035 Arg His Pro Arg Ala Tyr Thr Asn Lys Arg Leu Ala His Leu Thr 1040 1045 1050 Val Tyr Tyr Lys Leu Met Ala Arg Arg Ala Gln Val Pro Ser Ile 1055 1060 1065 Lys Asp Arg Ile Pro Tyr Val Ile Val Ala Gln Thr Arg Glu Val 1070 1075 1080 Glu Glu Thr Val Ala Arg Leu Ala Ala Leu Arg Glu Leu Asp Ala 1085 1090 1095 Ala Ala Pro Gly Asp Glu Pro Ala Pro Pro Ala Ala Leu Pro Ser 1100 1105 1110 Pro Ala Lys Arg Pro Arg Glu Thr Pro Ser Pro Ala Asp Pro Pro 1115 1120 1125 Gly Gly Ala Ser Lys Pro Arg Lys Leu Leu Val Ser Glu Leu Ala 1130 1135 1140 Glu Asp Pro Ala Tyr Ala Ile Ala His Gly Val Ala Leu Asn Thr 1145 1150 1155 Asp Tyr Tyr Phe Ser His Leu Leu Gly Ala Ala Cys Val Thr Phe 1160 1165 1170 Lys Ala Leu Phe Gly Asn Asn Ala Lys Ile Thr Glu Ser Leu Leu 1175 1180 1185 Lys Arg Phe Ile Pro Glu Val Trp His Pro Pro Asp Asp Val Thr 1190 1195 1200 Ala Arg Leu Arg Ala Ala Gly Phe Gly Ala Val Gly Ala Gly Ala 1205 1210 1215 Thr Ala Glu Glu Thr Arg Arg Met Leu His Arg Ala Phe Asp Thr 1220 1225 1230 Leu Ala 1235 18 1235 PRT herpes simplex 18 Met Phe Ser Gly Gly Gly Gly Pro Leu Ser Pro Gly Gly Lys Ser Ala 1 5 10 15 Ala Arg Ala Ala Ser Gly Phe Phe Ala Pro Ala Gly Pro Arg Gly Ala 20 25 30 Gly Arg Gly Pro Pro Pro Cys Leu Arg Gln Asn Phe Tyr Asn Pro Tyr 35 40 45 Leu Ala Pro Val Gly Thr Gln Gln Lys Pro Thr Gly Pro Thr Gln Arg 50 55 60 His Thr Tyr Tyr Ser Glu Cys Asp Glu Phe Arg Phe Ile Ala Pro Arg 65 70 75 80 Val Leu Asp Glu Asp Ala Pro Pro Glu Lys Arg Ala Gly Val His Asp 85 90 95 Gly His Leu Lys Arg Ala Pro Lys Val Tyr Cys Gly Gly Asp Glu Arg 100 105 110 Asp Val Leu Arg Val Gly Ser Gly Gly Phe Trp Pro Arg Arg Ser Arg 115 120 125 Leu Trp Gly Gly Val Asp His Ala Pro Ala Gly Phe Asn Pro Thr Val 130 135 140 Thr Val Phe His Val Tyr Asp Ile Leu Glu Asn Val Glu His Ala Tyr 145 150 155 160 Gly Met Arg Ala Ala Gln Phe His Ala Arg Phe Met Asp Ala Ile Thr 165 170 175 Pro Thr Gly Thr Val Ile Thr Leu Leu Gly Leu Thr Pro Glu Gly His 180 185 190 Arg Val Ala Val His Val Tyr Gly Thr Arg Gln Tyr Phe Tyr Met Asn 195 200 205 Lys Glu Glu Val Asp Arg His Leu Gln Cys Arg Ala Pro Arg Asp Leu 210 215 220 Cys Glu Arg Met Ala Ala Ala Leu Arg Glu Ser Pro Gly Ala Ser Phe 225 230 235 240 Arg Gly Ile Ser Ala Asp His Phe Glu Ala Glu Val Val Glu Arg Thr 245 250 255 Asp Val Tyr Tyr Tyr Glu Thr Arg Pro Ala Leu Phe Tyr Arg Val Tyr 260 265 270 Val Arg Ser Gly Arg Val Leu Ser Tyr Leu Cys Asp Asn Phe Cys Pro 275 280 285 Ala Ile Lys Lys Tyr Glu Gly Gly Val Asp Ala Thr Thr Arg Phe Ile 290 295 300 Leu Asp Asn Pro Gly Phe Val Thr Phe Gly Trp Tyr Arg Leu Lys Pro 305 310 315 320 Gly Arg Asn Asn Thr Leu Ala Gln Pro Arg Ala Pro Met Ala Phe Gly 325 330 335 Thr Ser Ser Asp Val Glu Phe Asn Cys Thr Ala Asp Asn Leu Ala Ile 340 345 350 Glu Gly Gly Met Ser Asp Leu Pro Ala Tyr Lys Leu Met Cys Phe Asp 355 360 365 Ile Glu Cys Lys Ala Gly Gly Glu Asp Glu Leu Ala Phe Pro Val Ala 370 375 380 Gly His Pro Glu Asp Leu Val Ile Gln Ile Ser Cys Leu Leu Tyr Asp 385 390 395 400 Leu Ser Thr Thr Ala Leu Glu His Val Leu Leu Phe Ser Leu Gly Ser 405 410 415 Cys Asp Leu Pro Glu Ser His Leu Asn Glu Leu Ala Ala Arg Gly Leu 420 425 430 Pro Thr Pro Val Val Leu Glu Phe Asp Ser Glu Phe Glu Met Leu Leu 435 440 445 Ala Phe Met Thr Leu Val Lys Gln Tyr Gly Pro Glu Phe Val Thr Gly 450 455 460 Tyr Asn Ile Ile Asn Phe Asp Trp Pro Phe Leu Leu Ala Lys Leu Thr 465 470 475 480 Asp Ile Tyr Lys Val Pro Leu Asp Gly Tyr Gly Arg Met Asn Gly Arg 485 490 495 Gly Val Phe Arg Val Trp Asp Ile Gly Gln Ser His Phe Gln Lys Arg 500 505 510 Ser Lys Ile Lys Val Asn Gly Met Val Asn Ile Asp Met Tyr Gly Ile 515 520 525 Ile Thr Asp Lys Ile Lys Leu Ser Ser Tyr Lys Leu Asn Ala Val Ala 530 535 540 Glu Ala Val Leu Lys Asp Lys Lys Lys Asp Leu Ser Tyr Arg Asp Ile 545 550 555 560 Pro Thr Tyr Tyr Ala Ala Gly Pro Ala Gln Arg Gly Val Ile Gly Glu 565 570 575 Tyr Cys Ile Gln Asp Ser Leu Leu Val Gly Gln Leu Phe Phe Lys Phe 580 585 590 Leu Pro His Leu Glu Leu Ser Ala Val Ala Arg Leu Ala Gly Ile Asn 595 600 605 Ile Thr Arg Thr Ile Tyr Asp Gly Gln Gln Ile Arg Val Phe Thr Cys 610 615 620 Leu Leu Arg Leu Ala Asp Gln Lys Gly Phe Ile Leu Pro Asp Thr Gln 625 630 635 640 Gly Arg Phe Arg Gly Ala Gly Gly Glu Ala Pro Lys Arg Pro Ala Ala 645 650 655 Ala Arg Glu Asp Glu Glu Arg Pro Glu Glu Glu Gly Glu Asp Glu Asn 660 665 670 Glu Arg Glu Glu Gly Gly Gly Glu Arg Glu Pro Glu Gly Ala Arg Glu 675 680 685 Thr Ala Gly Arg His Val Gly Tyr Gln Gly Ala Arg Val Leu Asp Pro 690 695 700 Thr Ser Gly Phe His Val Asn Pro Val Val Val Phe Asp Phe Ala Ser 705 710 715 720 Leu Tyr Pro Ser Ile Ile Gln Ala His Asn Leu Cys Phe Ser Thr Leu 725 730 735 Ser Leu Arg Ala Asp Ala Val Ala His Leu Glu Ala Gly Lys Asp Tyr 740 745 750 Leu Glu Ile Glu Val Gly Gly Arg Arg Leu Phe Phe Val Lys Ala His 755 760 765 Val Arg Glu Ser Leu Leu Ser Ile Leu Leu Arg Asp Trp Leu Ala Met 770 775 780 Arg Lys Gln Ile Arg Ser Arg Ile Pro Gln Ser Ser Pro Glu Glu Ala 785 790 795 800 Val Leu Leu Asp Lys Gln Gln Ala Ala Ile Lys Val Val Cys Asn Ser 805 810 815 Val Tyr Gly Phe Thr Gly Val Gln His Gly Leu Leu Pro Cys Leu His 820 825 830 Val Ala Ala Thr Val Thr Thr Ile Gly Arg Glu Met Leu Leu Ala Thr 835 840 845 Arg Glu Tyr Val His Ala Arg Trp Ala Ala Phe Glu Gln Leu Leu Ala 850 855 860 Asp Phe Pro Glu Ala Ala Asp Met Arg Ala Pro Gly Pro Tyr Ser Met 865 870 875 880 Arg Ile Ile Tyr Gly Asp Thr Asp Ser Ile Phe Val Leu Cys Arg Gly 885 890 895 Leu Thr Ala Ala Gly Leu Thr Ala Val Gly Asp Lys Met Ala Ser His 900 905 910 Ile Ser Arg Ala Leu Phe Leu Pro Pro Ile Lys Leu Glu Cys Glu Lys 915 920 925 Thr Phe Thr Lys Leu Leu Leu Ile Ala Lys Lys Lys Tyr Ile Gly Val 930 935 940 Ile Tyr Gly Gly Lys Met Leu Ile Lys Gly Val Asp Leu Val Arg Lys 945 950 955 960 Asn Asn Cys Ala Phe Ile Asn Arg Thr Ser Arg Ala Leu Val Asp Leu 965 970 975 Leu Phe Tyr Asp Asp Thr Val Ser Gly Ala Ala Ala Ala Leu Ala Glu 980 985 990 Arg Pro Ala Glu Glu Trp Leu Ala Arg Pro Leu Pro Glu Gly Leu Gln 995 1000 1005 Ala Phe Gly Ala Val Leu Val Asp Ala His Arg Arg Ile Thr Asp 1010 1015 1020 Pro Glu Arg Asp Ile Gln Asp Phe Val Leu Thr Ala Glu Leu Ser 1025 1030 1035 Arg His Pro Arg Ala Tyr Thr Asn Lys Arg Leu Ala His Leu Thr 1040 1045 1050 Val Tyr Tyr Lys Leu Met Ala Arg Arg Ala Gln Val Pro Ser Ile 1055 1060 1065 Lys Asp Arg Ile Pro Tyr Val Ile Val Ala Gln Thr Arg Glu Val 1070 1075 1080 Glu Glu Thr Val Ala Arg Leu Ala Ala Leu Arg Glu Leu Asp Ala 1085 1090 1095 Ala Ala Pro Gly Asp Glu Pro Ala Pro Pro Ala Ala Leu Pro Ser 1100 1105 1110 Pro Ala Lys Arg Pro Arg Glu Thr Pro Ser Pro Ala Asp Pro Pro 1115 1120 1125 Gly Gly Ala Ser Lys Pro Arg Lys Leu Leu Val Ser Glu Leu Ala 1130 1135 1140 Glu Asp Pro Ala Tyr Ala Ile Ala His Gly Val Ala Leu Asn Thr 1145 1150 1155 Asp Tyr Tyr Phe Ser His Leu Leu Gly Ala Ala Cys Val Thr Phe 1160 1165 1170 Lys Ala Leu Phe Gly Asn Asn Ala Lys Ile Thr Glu Ser Leu Leu 1175 1180 1185 Lys Arg Phe Ile Pro Glu Val Trp His Pro Pro Asp Asp Val Ala 1190 1195 1200 Ala Arg Leu Arg Thr Ala Gly Phe Gly Ala Val Gly Ala Gly Ala 1205 1210 1215 Thr Ala Glu Glu Thr Arg Arg Met Leu His Arg Ala Phe Asp Thr 1220 1225 1230 Leu Ala 1235 19 1235 PRT herpes simplex 19 Met Phe Ser Gly Gly Gly Gly Pro Leu Ser Pro Gly Gly Lys Ser Ala 1 5 10 15 Ala Arg Ala Ala Ser Gly Phe Phe Ala Pro Ala Gly Pro Arg Gly Ala 20 25 30 Gly Arg Gly Pro Pro Pro Cys Leu Arg Gln Asn Phe Tyr Asn Pro Tyr 35 40 45 Leu Ala Pro Val Gly Thr Gln Gln Lys Pro Thr Gly Pro Thr Gln Arg 50 55 60 His Thr Tyr Tyr Ser Glu Cys Asp Glu Phe Arg Phe Ile Ala Pro Arg 65 70 75 80 Val Leu Asp Glu Asp Ala Pro Pro Glu Lys Arg Ala Gly Val His Asp 85 90 95 Gly His Leu Lys Arg Ala Pro Lys Val Tyr Cys Gly Gly Asp Glu Arg 100 105 110 Asp Val Leu Arg Val Gly Ser Gly Gly Phe Trp Pro Arg Arg Ser Arg 115 120 125 Leu Trp Gly Gly Val Asp His Ala Pro Ala Gly Phe Asn Pro Thr Val 130 135 140 Thr Val Phe His Val Tyr Asp Ile Leu Glu Asn Val Glu His Ala Tyr 145 150 155 160 Gly Met Arg Ala Ala Gln Phe His Ala Arg Phe Met Asp Ala Ile Thr 165 170 175 Pro Thr Gly Thr Val Ile Thr Leu Leu Gly Leu Thr Pro Glu Gly His 180 185 190 Arg Val Ala Val His Val Tyr Gly Thr Arg Gln Tyr Phe Tyr Met Asn 195 200 205 Lys Glu Glu Val Asp Arg His Leu Gln Cys Arg Ala Pro Arg Asp Leu 210 215 220 Cys Glu Arg Met Ala Ala Ala Leu Arg Glu Ser Pro Gly Ala Ser Phe 225 230 235 240 Arg Gly Ile Ser Ala Asp His Phe Glu Ala Glu Val Val Glu Arg Thr 245 250 255 Asp Val Tyr Tyr Tyr Glu Thr Arg Pro Ala Leu Phe Tyr Arg Val Tyr 260 265 270 Val Arg Ser Gly Arg Val Leu Ser Tyr Leu Cys Asp Asn Phe Cys Pro 275 280 285 Ala Ile Lys Lys Tyr Glu Gly Gly Val Asp Ala Thr Thr Arg Phe Ile 290 295 300 Leu Asp Asn Pro Gly Phe Val Thr Phe Gly Trp Tyr Arg Leu Lys Pro 305 310 315 320 Gly Arg Asn Asn Thr Leu Ala Gln Pro Arg Ala Pro Met Ala Phe Gly 325 330 335 Thr Ser Ser Asp Val Glu Phe Asn Cys Thr Ala Asp Asn Leu Ala Ile 340 345 350 Glu Gly Gly Met Ser Asp Leu Pro Ala Tyr Lys Leu Met Cys Phe Asp 355 360 365 Ile Glu Cys Lys Ala Gly Gly Glu Asp Glu Leu Ala Phe Pro Val Ala 370 375 380 Gly His Pro Glu Asp Leu Val Ile Gln Ile Ser Cys Leu Leu Tyr Asp 385 390 395 400 Leu Ser Thr Thr Ala Leu Glu His Val Leu Leu Phe Ser Leu Gly Ser 405 410 415 Cys Asp Leu Pro Glu Ser His Leu Asn Glu Leu Ala Ala Arg Gly Leu 420 425 430 Pro Thr Pro Val Val Leu Glu Phe Asp Ser Glu Phe Glu Met Leu Leu 435 440 445 Ala Phe Met Thr Leu Val Lys Gln Tyr Gly Pro Glu Phe Val Thr Gly 450 455 460 Tyr Asn Ile Ile Asn Phe Asp Trp Pro Phe Leu Leu Ala Lys Leu Thr 465 470 475 480 Asp Ile Tyr Lys Val Pro Leu Asp Gly Tyr Gly Arg Met Asn Gly Arg 485 490 495 Gly Val Phe Arg Val Trp Asp Ile Gly Gln Ser His Phe Gln Lys Arg 500 505 510 Ser Lys Ile Lys Val Asn Gly Met Val Asn Ile Asp Met Tyr Gly Ile 515 520 525 Ile Thr Asp Lys Ile Lys Leu Ser Ser Tyr Lys Leu Asn Ala Val Ala 530 535 540 Glu Ala Val Leu Lys Asp Lys Lys Lys Asp Leu Ser Tyr Arg Asp Ile 545 550 555 560 Pro Ala Tyr Tyr Ala Ala Gly Pro Ala Gln Arg Gly Val Ile Gly Glu 565 570 575 Tyr Cys Ile Gln Asp Ser Leu Leu Val Gly Gln Leu Phe Phe Lys Phe 580 585 590 Leu Pro His Leu Glu Leu Ser Ala Val Ala Arg Leu Ala Gly Ile Asn 595 600 605 Ile Thr Arg Thr Ile Tyr Asp Gly Gln Gln Ile Arg Val Phe Thr Cys 610 615 620 Leu Leu Arg Leu Ala Asp Gln Lys Gly Phe Ile Leu Pro Asp Thr Gln 625 630 635 640 Gly Arg Phe Arg Gly Gly Gly Gly Glu Ala Pro Lys Arg Pro Ala Ala 645 650 655 Ala Arg Glu Asp Glu Glu Arg Pro Glu Glu Glu Gly Glu Asp Glu Asp 660 665 670 Glu Arg Glu Glu Gly Gly Gly Glu Arg Glu Pro Glu Gly Ala Arg Glu 675 680 685 Thr Ala Gly Arg His Val Gly Tyr Gln Gly Ala Arg Val Leu Asp Pro 690 695 700 Thr Ser Gly Phe His Val Asn Pro Val Val Val Phe Asp Phe Ala Ser 705 710 715 720 Leu Tyr Pro Ser Ile Ile Gln Ala His Asn Leu Cys Phe Ser Thr Leu 725 730 735 Ser Leu Arg Ala Asp Ala Val Ala His Leu Glu Ala Gly Lys Asp Tyr 740 745 750 Leu Glu Ile Glu Val Gly Gly Arg Arg Leu Phe Phe Val Lys Ala His 755 760 765 Val Arg Glu Ser Leu Leu Ser Ile Leu Leu Arg Asp Trp Leu Ala Met 770 775 780 Arg Lys Gln Ile Arg Ser Arg Ile Pro Gln Ser Ser Pro Glu Glu Ala 785 790 795 800 Val Leu Leu Asp Lys Gln Gln Ala Ala Ile Lys Val Val Cys Asn Ser 805 810 815 Val Tyr Gly Phe Thr Gly Val Gln His Gly Leu Leu Pro Cys Leu His 820 825 830 Val Ala Ala Thr Val Thr Thr Ile Gly Arg Glu Met Leu Leu Ala Thr 835 840 845 Arg Glu Tyr Val His Ala Arg Trp Ala Ala Phe Glu Gln Leu Leu Ala 850 855 860 Asp Phe Pro Glu Ala Ala Asp Met Arg Ala Pro Gly Pro Tyr Ser Met 865 870 875 880 Arg Ile Ile Tyr Gly Asp Thr Asp Ser Ile Phe Val Leu Cys Arg Gly 885 890 895 Leu Thr Ala Ala Gly Leu Thr Ala Val Gly Asp Lys Met Ala Ser His 900 905 910 Ile Ser Arg Ala Leu Phe Leu Ser Pro Ile Lys Leu Glu Cys Glu Lys 915 920 925 Thr Phe Thr Lys Leu Leu Leu Ile Ala Lys Lys Lys Tyr Ile Gly Val 930 935 940 Ile Tyr Gly Gly Lys Met Leu Ile Lys Gly Val Asp Leu Val Arg Lys 945 950 955 960 Asn Asn Cys Ala Phe Ile Asn Arg Thr Ser Arg Ala Leu Val Asp Leu 965 970 975 Leu Phe Tyr Asp Asp Thr Val Ser Gly Ala Ala Ala Ala Leu Ala Glu 980 985 990 Arg Pro Ala Glu Glu Trp Leu Ala Arg Pro Leu Pro Glu Gly Leu Gln 995 1000 1005 Ala Phe Gly Ala Val Leu Val Asp Ala His Arg Arg Ile Thr Asp 1010 1015 1020 Pro Glu Arg Asp Ile Gln Asp Phe Val Leu Thr Ala Glu Leu Ser 1025 1030 1035 Arg His Pro Arg Ala Tyr Thr Asn Lys Arg Leu Ala His Leu Thr 1040 1045 1050 Val Tyr Tyr Lys Leu Met Ala Arg Arg Ala Gln Val Pro Ser Ile 1055 1060 1065 Lys Asp Arg Ile Pro Tyr Val Ile Val Ala Gln Thr Arg Glu Val 1070 1075 1080 Glu Glu Thr Val Ala Arg Leu Ala Ala Leu Arg Glu Leu Asp Ala 1085 1090 1095 Ala Ala Pro Gly Asp Glu Pro Ala Pro Pro Ala Ala Leu Pro Ser 1100 1105 1110 Pro Ala Lys Arg Pro Arg Glu Thr Pro Leu His Ala Asp Pro Pro 1115 1120 1125 Gly Gly Ala Ser Lys Pro Arg Lys Leu Leu Val Ser Glu Leu Ala 1130 1135 1140 Glu Asp Pro Ala Tyr Ala Ile Ala His Gly Val Ala Leu Asn Thr 1145 1150 1155 Asp Tyr Tyr Phe Ser His Leu Leu Gly Ala Ala Cys Val Thr Phe 1160 1165 1170 Lys Ala Leu Phe Gly Asn Asn Ala Lys Ile Thr Glu Ser Leu Leu 1175 1180 1185 Lys Arg Phe Ile Pro Glu Val Trp His Pro Pro Asp Asp Val Ala 1190 1195 1200 Ala Arg Leu Arg Ala Ala Gly Phe Gly Ala Val Gly Ala Gly Ala 1205 1210 1215 Thr Ala Glu Glu Thr Arg Arg Met Leu His Arg Ala Phe Asp Thr 1220 1225 1230 Leu Ala 1235

Claims (26)

We claim:
1. A method of selecting compounds that inhibit herpes viruses comprising:
a) measuring IC50 of a compound of interest that inhibits a wild type herpes virus,
b) measuring IC50 of the same compound that inhibits a binding domain mutant herpes virus which is the same strain as the wild type herpes virus,
c) comparing IC50 of step a with IC50 of step b; and
d) selecting the compound of interest wherein the IC50 of step b is at least 3 times greater than the IC50 of step a.
2. A method of selecting compounds that inhibit herpes viruses comprising:
a) measuring IC50 of a compound of interest that inhibits a binding domain mutant herpes virus,
b) measuring IC50 of the same compound that inhibits a wild type herpes virus which is the same strain as the mutant herpes virus,
c) comparing IC50 of step a with IC50 of step b; and
d) selecting the compound of interest wherein the IC50 of step a is at least 3 times greater than the IC50 of step b.
3. The method of claim 1 or 2 wherein the herpes virus is HSV-1, HSV-2, HCMV, VZV, EBV, or HHV-8.
4. A method of selecting compounds that inhibit herpes viruses comprising:
a) measuring IC50 of a compound of interest that inhibits a wild type HSV-1,
b) measuring IC50 of the same compound that inhibits a binding domain mutant HSV-1 which is the same strain as the wild type herpes virus,
c) comparing IC50 of step a with IC50 of step b; and
d) selecting the compound of interest wherein the IC50 of step b is at least 3 times greater than the IC50 of step a.
5. A method of selecting compounds that inhibit herpes viruses comprising:
a) measuring IC50 of a compound of interest that inhibits a binding domain mutant HSV-1,
b) measuring IC50 of the same compound that inhibits a wild type herpes virus which is the same strain as the mutant HSV-1,
c) comparing IC50 of step a with IC50 of step b; and
d) selecting the compound of interest wherein the IC50 of step a is at least 3 times greater than the IC50 of step b.
6. The method of claim 4 or 5 wherein HSV-1 is HSV-1 KOS, HSV-1 F, HSV-1 DJL or HSV-1 Patton.
7. The method of claim 5 or 6 wherein the mutation of a wild type herpes virus to mutant herpes virus is at amino acid 823 from valine to alanine.
8. A method of selecting compounds that inhibit herpes viruses comprising:
a) measuring IC50 of a compound of interest that inhibits a wild type HSV-2,
b) measuring IC50 of the same compound that inhibits a binding domain mutant HSV-2 which is the same strain as the wild type herpes virus,
c) comparing IC50 of step a with IC50 of step b; and
d) selecting the compound of interest wherein the IC50 of step b is at least 3 times greater than the IC50 of step a.
9. A method of selecting compounds that inhibit herpes viruses comprising:
a) measuring IC50 of a compound of interest that inhibits a binding domain mutant HSV-2,
b) measuring IC50 of the same compound that inhibits a wild type herpes virus which is the same strain as the mutant HSV-2,
c) comparing IC50 of step a with IC50 of step b; and
d) selecting the compound of interest wherein the IC50 of step a is at least 3 times greater than the IC50 of step b.
10. The method of claim 8 or 9 wherein HSV-2 is HSV-2 MS, HSV-2 35D, or HSV-2 186.
11. A method of selecting compounds that inhibit herpes viruses comprising:
a) measuring IC50 of a compound of interest that inhibits a wild type HCMV,
b) measuring IC50 of the same compound that inhibits a binding domain mutant HCMV which is the same strain as the wild type herpes virus,
c) comparing IC50 of step a with IC50 of step b; and
d) selecting the compound of interest wherein the IC50 of step b is at least 3 times greater than the IC50 of step a.
12. A method of selecting compounds that inhibit herpes viruses comprising:
a) measuring IC50 of a compound of interest that inhibits a binding domain mutant HCMV,
b) measuring IC50 of the same compound that inhibits a wild type herpes virus which is the same strain of the mutant HCMV,
c) comparing IC50 of step a with IC50 of step b; and
d) selecting the compound of interest wherein the IC50 of step a is at least 3 times greater than the IC50 of step b.
13. The method of claim 8 or 9 wherein HCMV is AD169.
14. The methods of claims 1, 4, 8, or 11 wherein IC50 of step b is at least 5 times greater than the IC50 of step a.
15. The methods of claims 2, 5, 9, or 12 wherein IC50 of step a is at least 5 times greater than the IC50 of step b.
16. A method for selectively treating diseases caused by herpes viruses in a human host comprising administering a compound to a human in need of such treatment wherein said compound inhibits herpes viruses by interaction with the binding domain in the viral DNA polymerase.
17. A method for selectively inhibiting herpes viruses in a human host comprising administering a compound to a human in need of such treatment wherein IC50 of the compound that inhibits a binding domain mutant herpes virus is at lease 3 times greater than IC50 of the compound that inhibits a wild type herpes virus which is the same strain as the mutant herpes virus.
18. The method of claim 17 wherein IC50 of the compound that inhibits a binding domain mutant herpes virus is at lease 5 times greater than IC50 of the compound that inhibits a wild type herpes virus which is the same strain as the mutant herpes viruse.
19. The method of claim 17 wherein herpes viruses is HSV-1, HSV-2, HCMV, VZV, EBV, or HHV-8.
20. A compound for treating herpesviral infections in a human host wherein IC50 of the compound that inhibits a binding domain mutant herpes virus is at lease 5 times greater than IC50 of the compound that inhibits a wild type herpes virus which is the same strain as the mutant herpes virus.
21. A compound for treating herpesviral infections in a human host wherein said compound inhibits the herpesvirus by interacting with the binding domain in the viral DNA polymerase.
22. The herpesviral infection of claim 20 or 21 which is HSV-1, HSV-2, HCMV, VZV, EBV, or HHV-8 infection.
23. A compound for the inhibiting of herpesvirus DNA polymerases wherein passage of a wild type herpes virus in the presence of said compound results a change of the wild type HSV-1 polymerases at amino acid 823 from valine to alanine.
24. A compound for inhibiting herpesvirus DNA polymerases wherein passage of a wild type herpes virus in the presence of said compound results in a change of the wild type HCMV polymerases at amino acid 823 from valine to alanine and at amino acid 824 from valine to leuline.
25. A mutant herpesvirus DNA molecule having a nucleotide sequence selected from a group consisting of SEQ.ID.NO. 1; SEQ.ID.NO. 3; SEQ.ID.NO. 5; SEQ.ID.NO. 7; SEQ.ID.NO. 9; and SEQ.ID.NO. 11.
26. A mutant herpesvirus polymerase amino acid molecule having an amino acid sequence selected from a group consisting of SEQ.ID.NO. 2; SEQ.ID.NO. 4; SEQ.ID.NO. 6; SEQ.ID.NO. 8; SEQ.ID.NO. 10 and SEQ.ID.NO. 12.
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