CN1126438A - Method of combating acyclovir-resistant herpes simplex viral infections - Google Patents

Abstract

The present invention provides a method for treating acyclovir-resistant herpes infections in a mammal. The method comprises administering a peptide derivative or a combination of the peptide derivative and an antiviral nucleoside analog to the infected mammal. The peptide derivative used for the method is represented by the formula A-B-D-CH2CH{CH2C(O)R<1>}C(O)-NHCH{CR<2>(R<3>)COOH}C(O)-E.

Description

The method of the herpes simplex viral infections of combating acyclovir-resistant

The present invention relates to a kind of to the method for mammal treatment to the herpes infection of acycloguanosine tool resistance.This method comprises that using a kind of peptide derivant maybe combines administration with this peptide derivant with a kind of antiviral nucleoside analogue.

Acycloguanosine is a most popular medicine in the treatment herpes infection.But in recent years, the report number of times of relevant herpes infection to acycloguanosine tool resistance is rising always; Referring to, P.A.Chatis et al. for example, N.Engl.J.Med., 320,297 (1989) and S.Safrin, Res.Virol., 143,125 (1992).Recently, in the patient who suffers from serious human immunodeficiency virus (HIV) infection, observe the type that this tool resistance infects more and more.Behind a kind of patient's immunocompromised host in back, the result causes them to be highly susceptible to being subjected to herpes simplex virus (HSV) 1 type and the especially infection of 2 types.

The biochemical character of the HSV separated strain characteristic of tool acycloguanosine resistance is described receives very big concern; Referring to by C.S.Crumpacker, J.Am.Acad.Dermatol., the summary of 18190 (1988) the relevant anti-acycloguanosine HSV that deliver.In brief, a kind of or two kinds of enzymes of the Strain of tool resistance by encoding viral, functional variation has taken place in thymidine kinase (TK) or archaeal dna polymerase (POL).

When the cell that will infect wild type HSV contacts with acycloguanosine, viral TK and cellular enzymes mutually specific energy in a phosphorylation of catalysis acycloguanosine to a greater extent.This phosphorylation thing that will be produced by cellular enzymes changes into the triphosphoric acid acycloguanosine subsequently.The interaction that takes place between this latter's triphosphoric acid thing and the viral dna polymerase is easier to carry out than the interaction that takes place between it and the cell dna polymerase.Therefore, this triphosphoric acid thing has been owing to become a kind of replacement substrate of this viral enzyme, and can reduce the synthetic of viral DNA owing to have the effect of chain terminator.

Existing people has described two kinds of resistance mechanisms that relate to virus thymidine kinase: (1) selects thymidine kinase deficiency mutant, this mutant inductive enzymatic activity after infection is very low, and the mutant of (b) selecting to have the reformed thymidine kinase of its substrate specificity, this kinases can be with the thymidine phosphorylation and can not be with the acycloguanosine phosphorylation.

The third resistance mechanism relates to archaeal dna polymerase, and this resistance mechanism is to select the polymerase of its coding that the mutant that changes has taken place, and promptly the enzyme that should make a variation is to the deactivation tool resistance of triphosphoric acid acycloguanosine.

Based on above-mentioned resistance mechanism, the HSV separated strain to acycloguanosine tool resistance can be divided into: thymidine deficiency (TK ^D) strain, thymidine anomaly (TK ^A) strain or archaeal dna polymerase anomaly (POL ^A) strain.

Although existing people has reported that some successfully treat the case that anti-acycloguanosine HSV infects with antiviral agent phosphine formic acid, the frequency that anti-acycloguanosine type infects appearance is rising always.This phenomenon is the problem of common existence in HIV sufferers; Referring to K.S.Erlish etal., N.Engl.J.Med., 320,293 (1989) and S.Safrin, J.AcquiredImmun.Defic.Syndr., 5 (Suppl.1), s29 (1992).Therefore press for a kind of method of administering this phenomenon.

The invention provides a kind of the effective of anti-acycloguanosine herpes infection, relatively method of safety of being used for the treatment of.

This method comprises that use is by P.L.Beaulieu, R.Deziel, the peptide derivant of describing in careful patent application PCT/CA/93/0095 jointly that N.Moss and R.Plante submitted on March 12nd, 1993 should relate in careful patent application and use this compounds for treating herpes infection.But the known antiviral agent that can resist herpes infection effectively differs and resists the herpes simplex virus of tool acycloguanosine resistance surely effectively; For example referring to J.J.O ' Bnienand D.M.Campoli-Richards, Drugs 37,233 (1989), pp 252-253.Therefore find peptide derivant of the present invention to resist the herpes simplex virus of tool acycloguanosine resistance effectively be wonderful and be useful.

The invention provides a kind of method for the treatment of the herpes simplex viral infections of tool acycloguanosine resistance to mammal.This method comprises peptide derivant or its medicinal acceptable salt: the A-B-D-CH to the structural formula 1 of the herpesvirus effective dose of administration opposing tool acycloguanosine resistance ₂CH{CH ₂C (O) R ¹C (O)-NHCH{CR ²(R ³) COOH}C (O)-E

1

Wherein A is a phenylacetyl group, hydrocinnamoyl, (4-aminophenyl) propiono, (4-fluorophenyl) propiono, (4-hydroxyphenyl) propiono, (4-methoxyphenyl)-propiono, 2-(benzyl)-3-hydrocinnamoyl, 2-{ (4-fluorophenyl) methyl }-3-(4-fluorophenyl) propiono, 2-{ (4-methoxyphenyl) methyl }-3-(4-methoxyphenyl) propiono or benzylamino carbonyl; B is (N-Me)-Val or (N-Me)-Ile; Perhaps A and B are combined together to form a kind of saturated alkyl amino-carbonyl, it is selected from following groups: the butyl amino carbonyl, 1-Methylethyl amino carbonyl, 1-methyl-propyl amino carbonyl, 1-ethyl propyl amino carbonyl, 1,1-dimethyl ethyl butyl amino carbonyl, 1-ethyl-butyl amino carbonyl, 1-propyl group butyl amino carbonyl, 1-ethyl pentyl group amino carbonyl, 1-butyl amyl group amino carbonyl, 1-ethyl-butyl amino carbonyl, 2-ethyl pentyl group amino carbonyl, 1-methyl isophthalic acid-propyl group butyl amino carbonyl, 1-ethyl-1-propyl group butyl amino carbonyl, 1,1-dipropyl butyl amino carbonyl, (1-propyl group cyclopenta) amino carbonyl and (1-propyl group cyclohexyl) amino carbonyl; D is Val, Ile or Tbg; R ¹Be the 1-Methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 1,1-dimethyl propyl, 2,2-dimethyl propyl, cyclobutyl, cyclopenta, cyclohexyl, 1-methylcyclopentyl, NR ⁴R ⁵R wherein ⁴Be hydrogen or low alkyl group and R ⁵Be low alkyl group, perhaps R ⁴And R ⁵Form pyrrolidinyl (pyrrolidino) with nitrogen-atoms, piperidino (piperidino), morpholino (morpholino) or 4-methyl isophthalic acid-piperazinyl (4-methylpiperazino) in conjunction with them; R ²Be hydrogen and R ³Be methyl, ethyl, the 1-Methylethyl, 1, the 1-dimethyl ethyl, propyl group, 2-acrylic or benzyl carry R ²And R ³Carbon atom have (R)-configuration, perhaps R ²And R ³Each is methyl or ethyl, perhaps R naturally ²And R ³Form cyclobutyl with carbon atom, cyclopenta or cyclohexyl in conjunction with them; And E is NHR ⁶R wherein ⁶Be the 2-methyl-propyl, 2,2-dimethyl propyl, 1 (R), 2,2-trimethyl propyl group, 1,1,2,2-tetramethyl propyl group, 1 (R)-ethyl-2,2-dimethyl propyl, 2-(R, S)-methyl butyl, 2, the 2-dimethylbutyl, 3,3-dimethylbutyl, 1 (R), 2,2-trimethyl butyl, 1 (R), 3,3-trimethyl butyl, the 2-ethyl-butyl, 2,2-diethyl butyl, 2-ethyl-1 (R)-methyl butyl, 2-ethyl-2-methyl butyl, 1 (R)-ethyl-3, the 3-dimethylbutyl, 2,2-dimethyl amyl group, cis or trans-2-methylcyclohexyl, 2,2-Dimethylcyclohexyl or cyclohexyl methyl; Or E is NHCH (R ⁷)-Z wherein carries R ⁷Carbon atom have (S)-configuration, R ⁷Be 1, the 1-dimethyl ethyl, the 1-methyl-propyl, the 2-methyl-propyl, 2,2-dimethyl propyl or cyclohexyl methyl and Z are CH ₂OH, C (O) OH), C (O) NH ₂Or C (O) OR ⁸R wherein ⁸Be methyl, ethyl or propyl group.

The preferable methods that the present invention is used for the treatment of anti-acycloguanosine herpes-ness progenitalis infection comprises uses acceptable salt on structural formula 1 described peptide or its materia medica, and A is a hydrocinnamoyl in the structural formula 1,2-(benzyl)-3-hydrocinnamoyl or benzylaminocarbonyl; B is (N-Me) Val; D is Tbg; R ¹Be the 1-Methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 1,1-dimethyl propyl, 2,2-dimethyl propyl, cyclobutyl, cyclopenta, cyclohexyl or 1-methylcyclopentyl; R ²Be hydrogen and R ³Be methyl, ethyl, the 1-Methylethyl, propyl group or benzyl, and carry R ²And R ³Carbon atom have (R)-configuration, perhaps R ²And R ³Be separately methyl or ethyl, perhaps R separately ²And R ³Form cyclobutyl with carbon atom, cyclopenta or cyclohexyl in conjunction with them; And E is NHR ⁶R wherein ⁶Be 2, the 2-dimethyl propyl, 1 (R), 2,2-trimethyl propyl group, 1 (R)-ethyl-2, the 2-dimethyl propyl, 2,2 one dimethylbutyls or 1 (R)-ethyl-3,3-dimethylbutyl or E are NHCH (R ⁷)-Z wherein carries R ⁷Carbon atom have (S)-configuration, R ⁷Be 2,2-dimethyl propyl and Z are CH ₂OH, C (O) OH, C (O) NH ₂Perhaps C (O) OR ⁸R wherein ⁸Be methyl, ethyl or propyl group.

The another kind of method for optimizing that is used for the treatment of the herpes-ness progenitalis infection of anti-acycloguanosine comprises acceptable salt on the materia medica of using structural formula 1 described peptide derivant or this peptide derivant, wherein A in the structural formula 1 and B are combined together to form a kind of saturated alkyl amino-carbonyl, it is selected from following one group of group: 1-ethyl propyl amino carbonyl, 1-ethyl-butyl amino carbonyl, 1-propyl group butyl amino carbonyl, 2-ethyl pentyl group amino carbonyl, 1-methyl isophthalic acid-propyl group butyl amino carbonyl, 1-ethyl-1-propyl group butyl amino carbonyl, 1,1-dipropyl butyl amino carbonyl and (1-propyl group cyclopenta) amino carbonyl; And D, R ¹, R ², R ³With E all according to the definition in the last example.

Another aspect of the present invention relates to a kind of method for the treatment of anti-acycloguanosine herpesvirus infection to mammal, this method comprises on the structural formula 1 described peptide derivant of the kitchen rash aequum that can resist tool acycloguanosine resistance effectively or its materia medica that described mammal is given in acceptable salt associating dispenser on the acceptable salt and a kind of antiviral nucleoside analogue or its materia medica.

The antiviral nucleoside analogue that uses during administering drug combinations is that a kind of can enzymatic change (in body) be a kind of viral dna polymerase inhibitor of herpes dna AG14361, and/or changes a kind of material of replacing substrate of this enzyme into.This antiviral nucleoside analogue can be selected from known nucleoside analog.Preferred nucleoside analog of the present invention comprises acycloguanosine and its analog; The chemical compound of structural formula 2 for example:

R wherein ⁹Be hydrogen, hydroxyl or amino, or acceptable salt on the materia medica of this chemical compound.(R in the structural formula 2 ⁹This chemical compound is an acycloguanosine when being hydroxyl.)

The preferred antiviral nucleoside analogue of used according to the invention other comprises vidarabine, idoxene, trifluridine, gancyclovir, edoxudine, brovavir, fiac-itabine, penciclovir, famciclovir and rociclovir.

Accompanying drawing is described

The result that Fig. 1 and 2 graphic representation obtains when using potentiation in research process of isobole method proof, described research relates to the effect that shows when peptide derivant administering drug combinations with acycloguanosine and general formula 1 is used to resist the herpes simplex virus of tool acycloguanosine resistance.

Alternatively, structural formula 1 can be by following description

Being meant by corresponding a-amino acid from it, removes term " residue " hydroxyl and alpha-amino-group that individual hydrogen obtains later on the carboxyl when specially referring to a seed amino acid or amino acid derivativges.

Generally speaking, the abbreviation that is used to name aminoacid and protectiveness group in this article is to be foundation with IUPAC-IUB commission on Biochemical nomenclature content recommendation, referring to European Journ-al of Biochemistry 138,9 (1984).Val for example, Ile, Asp and Leu represent L-valine residue respectively, L-isoleucine residue, L-asparagicacid residue and L-leucine residue.

The main linear axes (being main chain) that is positioned at structural formula 1 peptide derivant (except end group A and (E's) Z, but comprises as E and is NHCH (R as defined herein ⁷Carry R during)-Z ⁷Carbon atom) on asymmetric carbon atom have the S configuration.But have except a kind of situation, promptly for carrying CH ₂C (O) R ¹The carbon atom of side chain and R wherein ¹For the low alkyl group of definition herein or during low-grade cycloalkyl.For this exceptional case, this carbon atom has the R configuration.

Be positioned on the side chain of aminoacid or deutero-amino acid residue, and be positioned at terminal A group and terminal E group (when E represents NHR as defined herein ⁶) on asymmetric carbon atom can have S or R configuration.

" " Pr and " Bu " code name are represented the alkyl group methyl respectively, ethyl, propyl group and butyl for Me, " Et ".

For example, code name " MeEt ₂C " and " EtPr ₂C " represent 1-ethyl-1-methyl-propyl and 1-ethyl-1-propyl group butyl group respectively.

Code name " Tbg " represented amino acid residue (S)-2-amino-3, the 3-acid dimethyl." γ MeLeu " represented amino acid residue (S)-2-amino-4,4-dimethyl valeric acid.Code name " γ MeLeucinol " representative (S)-2-amino-4, the 4-dimethyl pentanol is removed a hydrogen on this amylalcohol alpha-amido.

Other code name of Shi Yonging is in this article: (N-Me) Val represents (S)-3-methyl-2-(methylamino)-butanoic acid residue; (N-Me) Ile represents residue (S)-3-methyl-2 (methylamino) valeric acid; (N-Me ₃) Tbg represents residue (S)-2 (methylamino)-3,3-acid dimethyl; ASP (cyBu) represents residue (S)-alpha-amido-1-carboxyl Tetramethylene. acetic acid; And Asp (cyPn) expression residue (S)-alpha-amido-1-carboxyl Pentamethylene. acetic acid.

Term used herein " low alkyl group ", separately the form or the form that combines with other groups are meant straight chained alkyl group that contains 1-6 carbon atom and the branched alkyl group that contains 3-6 carbon atom, and comprise methyl, ethyl, propyl group, butyl, hexyl, the 1-Methylethyl, the 1-methyl-propyl, 2-methyl-propyl and 1,1-dimethyl ethyl.

Term used herein " 1-(low alkyl group)-(low-grade cycloalkyl) " is meant a kind of low-grade cycloalkyl group, and it carries a low-grade alkyl substituent on 1; For example, 1-ethyl cyclopropyl, 1-propyl group cyclopenta and 1-propyl group cyclohexyl.

Term used herein " low-grade cycloalkyl ", separately form or be meant the saturated cyclic hydrocarbon group that contains 3-6 carbon atom with other residue forms of combining comprises cyclopropyl, cyclobutyl, cyclopenta and cyclohexyl.

Term used herein " can be accepted carrier on the materia medica " and be meant to compose a kind of nontoxic of agent as active component, is generally inert material, and this excipient can't produce adverse effect to this active component.

Term used herein " the last acceptable carrier of physiology " is meant that a kind of of one or more non-toxic excipients can be by the excipient of the used for cosmetic that is subjected to, and described excipient can not react with the active component that is comprised or reduce its activity.

Term " effective dose " refers to the antiviral amount of measuring in advance of this antiviral agent, promptly is enough to resist effectively in vivo the amount of the reagent of viral organism body.

Term used herein " coupling agent " thus be meant the carboxyl that can make a seed amino acid or peptide and the dehydration coupling forms amido link between two kinds of reactants reagent takes place the free amine group of another kind of aminoacid or peptide.Equally, such reagent can make a kind of acid and a kind of alcohol coupling take place and form corresponding ester.These reagent are by activating carboxyl and impel or helping to take place the dehydration coupling.These coupling agents and activated group are described in the textbook of relevant chemistry of peptides usually; For example, E.Schroder and K.L.Lubke, " The Peptides ", Vo.1, Ac-ademic Press, New York, N.Y., 1965, pp 2-128, and K.D.Ko-pple, " Peptides and Amino acids ", W.A.Benjamin, Inc., NewYork, N.Y., 1966, pp 33-51.The example of coupling agent has the diphenylphosphine acyl azide, 1,1 '-carbonyl dimidazoles, dicyclohexyl carbodiimide, N-hydroxy-succinamide, the 1-hydroxyl-benzotriazole when perhaps having dicyclohexyl carbodiimide to exist.A kind of very feasible and useful coupling agent are at Tetrahedron Lett-ers by people such as B.Castro, 1219 (1975), also can be referring to D.Hudson, J.Org.Chem., 53,-three of 617 (1988) (benzotriazole-1-base oxygen) described-(dimethylamino) Phosphonium hexafluorophosphate can use this chemical compound separately or use in the presence of I-hydroxybenzotriazole.Also having another kind of very feasible and useful coupling agent is 2 (1H-benzotriazole-1-the yl)-N that can buy by commercial sources, N, N ' N '-tetramethylurea tetrafluoroborate.

Acceptable salt is a compounds of knowing in this area on antiviral nucleoside analogue that uses among the present invention and the materia medica thereof.As mentioned above, this compounds member is characterised in that the mode of the antivirus action of its mediation opposing herpesvirus, promptly realizes antiviral effect by suppressing viral dna polymerase in the body.The important member of this compounds acycloguanosine is arranged and by H.J.Schaeffer at United States Patent (USP) 4,199, describe in 574 (announcements on April 22nd, 1980), also can be referring to people such as H.J.Schaeffer, Nature (London), 272,583 (1978) and people such as T.A.Krenitsk, Proc.Natl.Acad.Sai.USA, 81,3209 (1984) the middle acycloguanosine analog of describing.R wherein ⁹During for hydroxyl, the chemical compound of structural formula 2 is " acycloguanosine ", chemical name 9-[(2-hydroxyl-ethyoxyl that it is known in addition) methyl] guanine.R wherein ⁹Structural formula 2 during for hydrogen respectively be called 6-descycl and 2-amino-9-[(2-hydroxyl-oxethyl) methyl] adenine; Work as R ⁹The chemical name of the chemical compound of general formula 2 is 2 when being amino, 6-diaminourea-9-[(2-hydroxyl-oxethyl) methyl] purine.

Work as R ⁹The chemical compound of structural formula 2 is with its tautomeric form 2-amino-1 when being hydroxyl, 9-dihydro-9-[(2-hydroxyl-oxethyl) methyl]-it is self-evident that 6H-purine-6-one form exists, and this chemical compound can be the mixture of two kinds of tautomeric forms, and the percentage ratio of various tautomers is decided according to the natural environment of this chemical compound in this mixture.But also has tautomeric form for other antiviral nucleoside analogues with enolization carbonyl.

Consider among the present invention that other anti-viral nucleosides that use comprise vidarabine (9-β-D-arabinofuranosyl base adenine sulfuric monohydrate), referring to people such as R.J.Whitley, N.Engl.J.Med., 307,971 (1982); Idoxene (2 '-deoxidation-5-ioduria glycosides), referring to W.H.Prusoff, Biochim.Biophys.Acta, 32,295 (1959); Trifluridine [2 '-deoxidation-5-(trifluoromethyl) uridnine], referring to C.Heidelberger, United States Patent (USP) 3,201,387 (issue August 17 nineteen sixty-five); Gancyclovir 9-[(1,3-dihydroxy-2-propoxyl group) methyl]-guanine, referring to J.P.Verheyden and J.C.Martin, United States Patent (USP) 4,355,032 (issue October 19 nineteen eighty-two); Edoxudine (5-ethyl-2 '-BrdU), referring to K.K.Gauri, US patent 3,553,192 (on January 5th, 1971 issued); Brovavir[(E)-5-(2-bromo vinyl)-2-BrdU], referring to people such as Y.Benoit, Eur.J.Pediatrics, 143,198 (1985); Fiacitabine (2 '-fluoro-deoxidation-5-ioduria glycosides), referring to people such as B.Leyla-nd-Jones, J.Infect.Dis., 154,430 (1986); Penciclovir (9-[4-hydroxyl-3-(hydroxyl-methyl)-butyl] guanine, referring to people such as S.E.Fowler, Br.J.Clin.Pharmacol., 28,2369 (1989); Famciclovir (9-[4-acetoxy-3-(acetoxy-methyl) butyl] adenine, referring to people such as P.A.V.Hodge, Antimicrob.Agents Chemotherap., 33,1765 (1989); And rociclovir (9-[(1,3-diisopropoxy-2-propoxyl group) methyl] adenine, referring to people such as E.Winklemann, Arzneim.-Forsch., 38,1545 (1988).

The method for preparing structural formula 1 peptide derivant

The method (for example titer phase method of couping amino acids residue and/or fragments of peptides) that use is generally adopted in synthesizing with peptide can prepare the peptide derivant of eliminant 1.Relevant description to these methods can be referring to for example, E.Schroder and K.Lubke, above-described textbook book series, " The Peptides:Analysis; Synthesis, Biology ", people such as E.Gross edit, Academic Press, New York, N.Y., 1979-1987, Vol.1-8, and J.M.Stewart and J.D.Young; " Solid Phase Pep-tide Synthesis ", 2nd ed., Pierce Chem.Co.Rockford, IL, USA, 1984.

The common trait of the method for introducing above that is used for synthetic peptide derivant is with the suitable various amino acid residues of protectiveness radical protection or deutero-amino acid residue (perhaps; if desired; the non-fragments of peptides of this peptide derivant) reactive side chain group; wherein said protectiveness group can prevent from described site chemical reaction to take place, until this blocking group of last removal.Another common ground is to protect its alpha-amido when an aminoacid or a segmental carboxyl react, and optionally removes its protectiveness group of this α-ammonia subsequently, so that the reaction of back takes place on this site.Also having a common trait is to protect C-terminal carboxyl group (if existence on this amino acid residue or the fragments of peptides with a suitable protectiveness group when initial; this carboxyl is the C-terminal functional groups of this peptide derivant); chemical reaction takes place in this protectiveness group prevention on this site, remove this protectiveness group after required peptide derivant sequence has been assembled.

The intermediate product of a key of the peptide of structural formula 1 is the intermediate product of structural formula 2.

W-D-CH ₂CH{CH ₂C (O) R ¹C (O) OH 2 wherein W be an alpha-amido protectiveness group, for example, tert-butyl-oxygen-carbonyl (BOC), benzyl-oxygen-carbonyl (Z) or fluorenes-9-ylmethoxy carbonyl (Fmoc), and D and R ¹By the content that defines herein.By adopting Michael (Michael) addition process with structural formula W-D-CH ₂C (O) OCH ₂CH=CH ²Allyl ester be added to structural formula (low alkyl group)-OC (O) CH=CHC (O) R ¹The fumaroyl derivant on to obtain structural formula be W-D-CH{C (O)-OCH ₂CH=CH ₂CH{CH ₂C (O) R ¹The Michael-adduct of C (O) O-(low alkyl group), thereby can prepare this chemical compound.

After this, according to R.Deziel, Tetrahedron Letters, 28,4371 (1987) method is handled this chemical compound that obtains with four triphenylphosphine palladiums and triphenylphosphine in the presence of pyrrolidine, this processing effect causes allyl ester generation hydrolysis and follow decarboxylate obtaining the crucial corresponding Arrcostab of intermediate product.For example sodium hydroxide or Lithium hydrate exist and issue unboiled water and separate the mixture that obtains the key intermediate species diastereomer this a kind of ester in back at alkali.

Another kind of adoptable method is to adopt the following step to prepare the key intermediate species shown in the general formula 2, and wherein W and D are according to the top content of middle definition for example, and R ¹Be NR ⁴R ⁵R wherein ⁴And R ⁵Each is low alkyl group or R naturally ⁴And R ⁵The nitrogen-atoms that combines with them forms pyrrolidinyl, piperidino, morpholino or 4-methyl isophthalic acid-piperazinyl: the condition that adopts the michael reaction generation is with structural formula BzlOC (O) CH=CHC (O) OBzl{ (E)-fumaroyl derivant of 2-butylene diacid dibenzyl ester and the structural formula W-D-CH that introduces above ₂C (O) O-CH ₂CH=CH ₂The allyl ester of (wherein W and D are by defining content herein) reacts and obtains the corresponding structure formula is W-D-(RS)-CH{CH{C (O) OBzl}CH ₂C (O) OBzl}C (O)-OCH ₂CH=CH ₂Michael-adduct.Handling this a kind of adduct in back (Deziel, the vide document is the same) with four triphenylphosphine palladiums and triphenylphosphine in the presence of pyrrolidine, to obtain structural formula be W-D-CH ₂-(R, S)-CH{CH ₂The corresponding dibenzyl ester of C (O) OBzl}C (O) OBzl.Hydrogenolysis taking place on carbon in the presence of palladium dydroxide subsequently, obtain corresponding dicarboxylic acids, it is heated with excessive acetic anhydride via and is converted into corresponding anhydride.In the presence of pyridine this anhydride and suitable secondary amine are reacted, obtain the mixture of various positions isomer (re-gioisomer), structural formula is W-D-CH in this mixture ₂-(RS)-CH{CH ₂C (O) NR ⁴R ⁵}-C (O)-OH (wherein W, D and NR ⁴R ⁵Be according to defining content herein) needed isomer comprises advantage.This product is converted into benzyl ester, and adopts high performance liquid chromatography that the position isomer mixture that obtains is separated, obtain required isomer as the advantage product.Hydrogenation generation key intermediate species structural formula 2 takes place in carbon atom with this product in the presence of palladium dydroxide subsequently, and wherein W and D press definition herein, R ¹And NR ⁴R ⁵Also press this paper definition.

Therefore; in general; the employing order is a peptide sequence; with suitable aminoacid or deutero-amino acid residue; and the non-fragments of peptides of this peptide (for example key intermediate species) (if desired; can protect these fragments suitably) progressively coupling, and after progressively coupling is finished, eliminate all protecting groups because of (if existence), thus prepare the peptide of structural formula 1.More specifically method will be hereinafter embodiment partly describe.

Can prepare acceptable salt form on the therapeutics of eliminant 1 peptide of the present invention.The situation that contains the residue of tool alkali effect for a kind of specific peptide, the example of such salt of this alkali has for example acetate of those acylates, lactate, succinate, methane sulfonates or right-toluene fulfonate, and polymeric acid for example tannate or carboxymethyl cellulose salt, can also be inorganic acid salt as with halogen acids, for example hydrochloric acid, or sulphuric acid, or the formed salt of phosphoric acid.If desired, can be by people such as R.A.Boissonnas, Helv.Chim.Acta, 43,1849 (1960) methods of describing change specific acid-addition salts into another kind of acid-addition salts with suitable ion exchange resin treatment, for example a kind of nontoxic, acceptable salt on the materia medica.

Contain the situation that one or more free carboxies are rolled into a ball for a kind of specific peptide, the example of the described salt of this carboxyl has sodium salt, potassium salt or calcium salt, and perhaps organic alkali salt is for example with triethylamine or the formed salt of N-methylmorpholine.

The herpes activity

The antiviral activity that the peptide derivant that adopts display structure formula 1 can prove this chemical compound to inhibiting biochemical, the microbiology and the biological method of tool acycloguanosine resistance herpes simplex virus 1 type and 2 types (HSV-1 and HSV-2).

The peptide derivant that is used for certification structure formula 1 is a cell culture technology to the inhibiting method of virus replication, referring to for example, and people such as T.Spector, Proc.Natl.Acad.Sci.USA, 82,4254 (1985).

Available laboratory animal proves the therapeutic effect of this peptide derivant, serve as according to the detection method of implementing for example by carry out the antiviral drugs detection with the mouse model of suffering from the inductive disease of eye of herpes simplex virus, referring to people such as C.R.Brandt, J.Virol.Meth., 36,209 (1992) describe.

When acceptable salt is used as a kind of antiviral agent on a kind of peptide derivant of the present invention or its a kind of therapeutics, it can be contained in a kind of excipient to homoiothermic animal people for example, pig or horse part or whole body administration, this excipient contains on one or more materia medicas can accept carrier, and each composition ratio of this medicament is by convention decision biology of dissolubility and chemical property and the selected route of administration and the standard of this peptide derivant.For topical, this derivant can be able to be accepted excipient on materia medica and prepare, active component content is 0.1-5%, is preferably 0.5-2%.This medicament can be solution, emulsifiable paste or lotion.

For the whole body administration, the peptide derivant of structural formula 1 and pharmaceutically acceptable excipient or carrier can be combined into composition forms and give patient oral or intravenous injection, subcutaneous injection or intramuscular injection.During drug administration by injection, preferably use this peptide to be dissolved in the solution that a kind of aseptic water excipient forms, this excipient also can contain other solutes such as buffer agent or antiseptic and be used to keep acceptable salt on the materia medica of q.s of this solution isotonicity or the glucose of q.s.

The appropriate excipients of the medicament that is used for introducing above or carrier are described in the classical pharmacy textbook, for example " Remington ' s Pharmaceutical Sciences ", 18th ed, Mack Publishing Company, Easton, Penn., 1990.

The dosage of this peptide derivant changes with administering mode and selected specific active agent.In addition, also change with the specific diseased individuals for the treatment of.Usually, dosage is and increases by a small margin during begin treatment, up to the maximum efficiency that reaches under this condition.In general, optimal be with this peptide derivant with a certain concentration level administration, antiviral effectively usually when this concentration level, but can not produce any harmful or deleterious side effect.

Use this medicine as for the part, the suitable topical preparation of this peptide derivant is applied directly to the affected area of health, skin for example, eyes or oral cavity or genital atriium position, the amount foot of this medicament of using is in covering this affected area.This process should repeat, and for example repeats every 4-6 hour once up to the damage recovery from illness.

As for systemic administration, with structural formula 1 described peptide derivant with every day per kilogram of body weight use the dosed administration of 1.0mg-10mg, although the variation that can occur introducing previously.But optimal dosage level is the scope of every kg body weight use every day 1.0mg-5mg, thereby obtains effective result.

Though above disclosed preparation demonstrates it when being used for the treatment of herpesvirus infection be a kind of effective and comparatively safe medicament, do not get rid of these preparations and other antiviral agents or reagent reasonably can be obtained more effective result during administration simultaneously.Such antiviral drugs or reagent comprise previously described anti-viral nucleoside, antiviral surface-active agents or antiviral interferon for example by S.S.Asculai and F.Rapp at United States Patent (USP) 4,507, disclosed content in 281 (on March 26th, 1985).

More particularly, for the herpesvirus infection that adopts the tool of medication treatment simultaneously acycloguanosine resistance, having been found that peptide derivant with a kind of antiviral nucleoside analogue and structural formula 1 combines when using can strengthen the herpes activity of this analog synergistically, and does not follow the increase of toxic and side effects.Therefore, the invention provides a kind of pharmaceutical composition for the treatment of the herpes infection of tool acycloguanosine resistance to mammal, this pharmaceutical composition contains on a kind of materia medica or veterinarily acceptable carrier, and acceptable salt on a kind of antiviral nucleoside analogue of effective dose or its materia medica, with the compositions of acceptable salt on the peptide derivant of structural formula 1 or its materia medica.

When term " cooperative effect " is used to represent that the antiviral of compositions of peptide derivant of above defined nucleoside analog and structural formula 1 or herpes are active, it be meant this antiviral or herpes effect greater than two kinds of predetermined separate constituents in conjunction with after addition.

With the herpes infection of combination treatment tool acycloguanosine resistance of the present invention the time, said composition is contained in contains in the excipient that to accept carrier on one or more materia medicas to homoiothermic animal people for example, pig or horse administration, each components in proportions is by the dissolubility and the chemical characteristic of the peptide derivant of this nucleoside analog and structural formula 1 in the said composition, and selected route of administration, classical convention decision biology depends on that also can the relative amount of these two kinds of active component provide collaborative antiviral effect.In a kind of preferable methods, with the said composition topical.For example, acceptable excipient on these two kinds of active agents (being acceptable salt on the peptide derivant of antiviral nucleoside analogue and structural formula 1 or its therapeutics) and the materia medica is mixed with the solution form, Emulsion form, cream form or lotion form.Can contain acceptable salt on nucleoside analog that weight percent is 0.01-1.0% or its therapeutics in this preparation, and contain acceptable salt on the peptide derivant of structural formula 1 of the 0.05-1% weight of having an appointment or its therapeutics.

Under any circumstance, these two kinds of active agents all are present in this pharmaceutical composition with the amount that collaborative herpes effect can be provided.

The following examples further describe the present invention.Described temperature is meant Celsius temperature.Solution percentage ratio or ratio are represented the relative ratio of volume and volume, except as otherwise noted.NMR (Nuclear Magnetic Resonance) spectrum is by Bruker 200MHz or 400MHz spectrometer record (having put down in writing 400MHz spectrum in beginning); The chemical shift of being write down (δ) is meant proportion in 1,000,000.The abbreviation of Shi Yonging comprises Boc in an embodiment: uncle-butoxy carbonyl; Bu: butyl; Bzl: benzyl; DMF: dimethyl formamide; Et: ethyl; EtoH: ethanol; EtOAc: ethyl acetate; Et ₂O: diethyl ester; Me: methyl; MeOH: methanol; Pr: propyl group; TLC: thin layer chromatography; THF: oxolane.

Embodiment 1

The global schema of coupling reaction

Also can be referring to people such as R.Knorr., Tetrahedron Letters, 30,1927 (1989) .}

With first kind of reactant, promptly a kind of unhindered amina (or its hydrochlorate) is dissolved in CH ₂Cl ₂Or in the acetonitrile, this solution is cooled to 4 ℃.In the presence of nitrogen, 4 normal N-methylmorpholines are added in the solution of this vibration.After 20 minutes, adding 1 normal second kind of reactant is a kind of free carboxy acid, and 1.05 normal coupling agents.For above-mentioned purpose, practical and effectively coupling reagent be (benzotriazole-1-base oxygen) three-(dimethyl amine) phosphine hexafluorophosphates or 2-(1H-benzotriazole-1-yl)-N preferably, N, N ', N ' ,-tetramethylurea tetrafluoroborate.Monitor this reaction process with the TLC method.After reaction is finished, vapourisation under reduced pressure CH ₂Cl ₂(or acetonitrile).Residue is dissolved in EtOAc.Use the 1N aqueous citric acid solution, 10%Na ₂CO ₃Aqueous solution and saline wash this solution successively.Under reduced pressure with organic facies drying (MgSO ₄), filter and be concentrated into dried.According to the poly-chromatography (fl-ash chromatography technique) of the urgency of Still people such as {, J.Org.Chem., 43,2923 (1978) } W.C.Still at silica gel (SiO ₂) last this residue of purification.

Embodiment 2

Intermediate product H-Asp (cyPn) (Bzl)-NH-(S)-CH (CH ₂CMe ₃) CH ₂The preparation of OBzl

(a) (s)-α-azido-1-{ (benzyloxy } carbonyl }-Pentamethylene. acetic acid: according to the adminicle that utilizes Evan (referring to people such as D.A.Evans, J.Amer.Chem.Soc., 112,4011 (1990)] asymmetric Azide method, and people such as M.N.Aboul-Enein, Pharm.Acta Helv., 55,50 (1980) description, by 2-oxo spiral [4.4] nonane-1, the 3-diketone prepares this chemical compound.

More particularly, exist at argon, under-40 ℃ of conditions with butyl lithium (469ml, 1.6M hexane solution 750mmol) dropwise is added to chirality adminicle 4 (S)-(1-Methylethyl)-2-oxazolidone (4 (S)-(1-methylethyl)-the 2-Oxazolidinone) { 96.8g that is present among the anhydrous THF, 750mmol is by L.N.Pridgen and J.Prol., J.Org.Chem., 54,3231 (1989) } in.This mixture was shaken 30 minutes at-40 ℃, be cooled to-78 ℃ then.With 2-oxo spiral [4.4] nonane-1, the 3-diketone is added drop-wise in this refrigerative mixture.Then this mixture is vibrated 1 hour at 0 ℃.After this, 20% (w/v) aqueous citric acid solution (600ml) is added in this mixture.Organic facies is separated, used the EtOAc aqueous phase extracted.To merge the organic facies salt water washing of getting up, dry (MgSO ₄) and under reduced pressure, concentrate, obtain 3-{2-(1-carboxyl-cyclopenta)-1-oxoethyl of a kind of pink solid (300g) form) }-4 (S)-(1-Methylethyl)-2-oxazolidone.

This a kind of pink solid (about 750mmol) is dissolved in the acetonitrile (1 liter).(750mmol) with 1, (114 grams, 112ml 750mmol) is added in this solution 8-diazabicyclo [5.4.0]-7-hendecene for 128.3 grams, 89.2ml with benzyl bromide a-bromotoluene.This mixture is vibrated 16 hours in argon.Under reduced pressure, remove volatile matter.Residue is dissolved in H ₂Among the O/EtOAc.Isolate organic facies, with 10% (w/v) aqueous citric acid solution and salt water washing, dry (MgSO ₄) and under reduced pressure, be concentrated into drying, obtain a kind of oil.Should from hexane/EtOAc, crystallize out the corresponding benzyl ester that obtains a kind of white solid (204 grams, 73%) form by oil.

(70 grams, 187mmol) solution that is dissolved among the anhydrous THF (200ml) is cooled to-78 ℃ with the chemical compound that obtains at last.The potassium 1,1,1,3,3 that will contain 6% (w/v) cumene, (286ml, 0.66M THF solution 189mmol) joined in this cooling solution in 15 minutes the 3-hexamethyldisiloxane.This mixture is vibrated 45 minutes at-78 ℃.To be dissolved in the Azide 2,4 among the anhydrous THF (100ml), and 6-triisopropyl benzenesulfonic acid (67 grams 216mmol) once add in this cooling mixture, follow after two minutes, the adding glacial acetic acid (50ml, 860mmol).With this mixture heating, 35-45 ℃ of vibration 1 hour.Under reduced pressure, remove volatile matter.Should grind by the yellow residue with hexane/EtOH (4: 1,1.7 liters).The white solid that obtains is collected on the filter paper.With filtrate and SiO ₂(the 230-240 order sieves) mixes.Under reduced pressure, remove volatile matter, and under reduced pressure with the solid that obtains 35 ℃ of dryings so that remove cumene.Then residual solid is placed a SiO ₂In the pillar.With 9: 1 hexanes-EtOAc eluting residual solid and SiO ₂, and obtain 3-{{2 (S)-azido-1-oxo-2-{1-{ (phenyl methoxyl group) carbonyl after eluent concentrated } cyclopenta }-ethyl }-4 (S)-(the 1-Methylethyl }-2-oxazolidone (66 grams, 86%).

(13.42 grams 32.4mmol) are dissolved in THF/H with the chemical compound that obtains at last ₂(3: 1,608ml) solution in was cooled to 0 ℃ to O.With H ₂O ₂/ H ₂O (3: 7,16.3ml, the H of 518mmol ₂O ₂) be added in this cooling solution; Add LiOHH subsequently ₂O (2.86 grams, 68.2mmol).This mixture 0 ℃ of vibration 45 minutes, is used aqueous solution (400ml) quencher of the sodium sulfate of 10% (w/V) then.Adding NaHCO ₃(1.93 gram) concentrates this mixture under reduced pressure afterwards.By continuous extraction (NaHCO ₃Aqueous solution/chloroform) reclaimed the chirality adminicle in 20 hours.After this, this water is cooled to 0C and adds dense HCl and make it be acid, extract with EtOAc then.With salt water washing extract, dry (MgSO ₄) and under reduced pressure, concentrate the required compound that obtains white solid (8.2 grams, 84%) form.This chemical compound ¹H NMR (CDCl ₃) result shows: δ 1.6-1.8 (m, 5H), 1.95-2.05 (m, 2H), 2.20-2.30 (m, 1H), 4.55 (s, 1H), 5.12 (s, 2H) and 7.4 (m, 5H).

In (C) of present embodiment part, used this chemical compound.

(b) NH ₂-(S)-CH (CH ₂CMe ₃) CH ₂OBzl: according to A.Giannis and K.Sandh-off, Angew.Chem.Int.Ed.Engl., 28,218 (1989) method LiBH ₄/ Me ₃SiCl reduction H-γ MeLeu-OH obtains amino alcohol NH ₂-(S)-CH (CH ₂CMe ₃) CH ₂OH.In nitrogen, and a chemical compound after inciting somebody to action under 4 ℃ of conditions (812mg, 6.2mmol), triethylamine (659mg, 6.51mmol) and the di-t-butyl heavy carbonate (1.42 grams 6.51mmol) were dissolved in the mixture vibration that obtains among the anhydrous THF (15ml) 15 minutes, at room temperature vibrated 4 hours then.Under reduced pressure, THF is evaporated.Residue is dissolved among the EtOAc.Use 10% aqueous citric acid solution, 5%NaHCO ₃Aqueous solution and this solution of salt water washing.With this organic facies drying (MgSO ₄) and under reduced pressure, be concentrated into dried.By anxious poly-chromatography (SiO ₂, eluent hexane-EtOAc, 2: 1) and this residue is carried out purification.Obtain Boc-NH-(S)-CH (CH ₂CMe ₃)-CH ₂OH (1.23 grams, 86%).

Two sulphuric acid TBuAs (106mg) and 50%MaOH aqueous solution (3ml) are joined Boc-NH-(S)-CH (CH successively ₂CMe ₃) CH ₂(1.23 grams are in benzyl chloride 5.35mmol) (13ml) solution for OH.The mixture that obtains is vibrated 90 minutes at 35-40 ℃.With the EtOAc dilution, and use H ₂O and salt water washing.With organic facies drying (MgSO ₄) and under reduced pressure, be concentrated into drying regime.Residue is dissolved in the hexane.This solution is injected into SiO ₂In the post.With this post of hexane eluting,, use hexane-EtOAc (2: 1) to be eluted to then and obtain Boc-NH-(S)-CH (CH so that remove benzyl chloride ₂CMe ₃) CH ₂OBzl.This back one chemical compound ¹H NMR (CDCl ₃) result show δ 0.95 (S, 9H), 1.42 (S, 9H), 1.30-1.55 (m, 2H), 3.42 (d, J=4Hz, 2H), 3.88 (wide, 1H), 4.54 (m, 3H), 7.23-7.4 (m, 5H).(1.28 grams 3.99mmol) are dissolved in the 6N HCl/ diox (10ml) with this back one chemical compound.At nitrogen, vibration is 45 minutes under 4 ℃ of conditions with this solution.Solvent evaporated obtains the hydrochlorate (1.05 gram) of required compound.In next joint of present embodiment, will use without this chemical compound that is further purified.

(c) title compound of present embodiment:, utilize the NH in the previous section according to the coupling step of embodiment 1 ₂-(S)-CH (CH ₂CMe ₃) CH ₂The hydrochlorate of OBzl is as first reactant, with (S)-α-nitrine-1-{ (phenyl the methoxyl group)-carbonyl in present embodiment (a) joint } Pentamethylene. acetic acid is as second reactant, obtain N-{ (S)-1-benzyloxymethyl-3, the 3-dimethylbutyl }-(S)-and α-nitrine-1-{ (benzyloxy) carbonyl } the Pentamethylene. acetamide.Adopt people such as N.Maiti, Tetrahedron Letters, 27,1423 (1986) method obtains the chemical compound that obtains the title compound of present embodiment with stannic chloride (II) reduction in MeOH.This chemical compound ¹H NMR (CDCl ₃) analysis result demonstration: δ 0.98 (s, 9H), 1.22-2.25 (m, 12H), 3.4 (d, J=4 Hz, 2H), 3.64 (s, 1H), 4.18 (wide m, 1H), 4.52 (s, 2H), 5.12 (s, 2H), 7.18 (d, J=7Hz, 1H), 7.22-7.38 (wide m, 10H).

Embodiment 3

Intermediate product Boc-Tbg-CH ₂-(RS)-CH (CH ₂C (O) CMe ₃) preparation of C (O) OH:

(a) magnesium salt of malonic acid allyl monoesters: in reflux, with 2,2-dimethyl-1,3-diox-4, the 6-diketone (100 grams, 0.69mol) and 1-propenol-3 (47ml 0.69mol) is dissolved in the solution that benzene (800ml) obtains and heated 24 hours.Under reduced pressure, solvent evaporation is fallen.Residue distilled under reduced pressure obtain malonic acid allyl monoesters (71 grams, 71%, boiling point 123-127 °/2.7mmHg).(71 grams 0.48mol) are dissolved among the anhydrous THF (300ml) with this ester.(28.5 grams 0.245mol) join in this solution with Diethoxymagnesium.Down this mixture is vibrated 4 hours at argon and room temperature (20-22 ℃).Vapourisation under reduced pressure falls solvent, uses Et ₂O grinds this residue and obtains a kind of sepia solid.This solid abrasive become molecule and under reduced pressure drying obtain required magnesium salt (56 grams, 73%).In next joint of present embodiment, use this salt.

(b) Boc-Tbg-CH ₂C (O) OCH ₂=CH ₂: (15 restrain, and 64mmol) are dissolved in the solution in the anhydrous acetonitrile (150ml) adding 1, and (12.6 restrain the 1-carbonyl dimidazoles, 78mmol) to Boc-Tbg-OH.Under argon, room temperature condition, with this mixture vibration 2 hours.With the magnesium salt of malonic acid allyl ester (24 grams, 78mmol) and 4-(N, N-dimethyl amine) pyridine (100mg) join in this mixture.This mixture was heated in reflux 1 hour, at room temperature vibrated then 18 hours.After this, this mixture is concentrated under reduced pressure.Residue is dissolved among the EtOAc (300ml).With 10% aqueous citric acid solution (2 * 100ml) and saline (2 * 100ml) wash this solution, dry (MgSO ₄) and be evaporated to dried.With anxious poly-chromatography (flash chromatography) (SiO ₂, eluent: hexane-EtOAc, 9: 1) and this residue of purification obtains required allyl ester (19.4 gram, 96%), and this ester is a kind of brown oil.Should the oil crystallization after continuing for some time.Carry out ¹H NMR (CDCl ₃) analyze and find that this chemical compound exists with ketone group-enol tautomer mixed form in chloroform, both relatively is 3: 1, δ 0.96 (s, 9H, enol form), 1.04 (s, 9H), 1.46 (s, 9H), 3.65 (s, 2H), 3.90 (d, J=8.5Hz, 1H, enol forms), 4,20 (d, J=7.5Hz, 1H), 4.65 (m, 2H), 5.10 (wide d, J=7.5 Hz, 1H), 5.20-5.40 (m, 2H), and 5.80-6.05 (m, 1H), 12 (s, 1H enol forms).In (d) of present embodiment joint, use this allyl ester.

(c) (E)-5,5-dimethyl-4-oxo-2-hexenoic acid ethyl ester: according to people such as S.Ma-nfredini, Tetrahedron Letters, 29,3997 (1988) method prepares this ethyl ester.Should the anxious poly-chromatography (SiO of butyrous coarse products ₂, eluent: hexane) purification obtains the required acetic acid of yellow oil form.This chemical compound ¹H NMR (CDCl ₃) analysis result be δ 1.21 (s, 9H), 1.33 (t, J=7.5Hz, 3H), 4.28 (q, J=7.5Hz, 2H), 6.78 (d, J=15.5Hz, 1H), 7.51 (d, J=15.5Hz, 1H).In next joint of present embodiment, use this ethyl ester.

(d) title compound of present embodiment: with the Boc-Tbg-CH that describes in present embodiment (b) chapters and sections ₂C (O) OCH ₂=CH ₂(0.67 gram 2.1mmol) is dissolved in the presence of argon among the anhydrous THF (25ml).(60% oil suspension, 0.095 gram 2.4mmol) is added in this solution with sodium hydride.This mixture was at room temperature vibrated 30 minutes.With (E)-5 of describing in present embodiment (c) joint, (0.4 35 grams 2.36mmol) add this mixture to 5-dimethyl-4-oxo-2-hexenoic acid ethyl ester.Vibrate this reactant mixture and finish (about 6 hours) up to judging to react by the TLC monitoring.After this, with 10% aqueous citric acid solution this mixture that quenches.Under reduced pressure, remove THF, with concentrate EtOAc (3 * 25ml) extractions that obtain.Wash this extract with water, dry (MgSO ₄) and under reduced pressure, concentrate.With the anxious poly-chromatography (SiO of residue ₂, eluent: hexane-EtOAc, 9: 1) and purification obtains corresponding michael reaction adduct (Michael reaction adduct) (1.06 gram, 100%).

By following method this Michael-adduct is converted into Boc-Tbg-CH ₂-(RS)-CH (CH ₂C (O) CMe ₃) C (O) OH: in argon with four triphenylphosphine palladiums (0) (0.20 gram, 0.18mmol) and triphenylphosphine (0.060 restrains, and 0.23mmol) is dissolved in CH ₂Among the Cl (10ml).(20ml) joins in the solution with acetonitrile, and this solution is cooled to 0 ℃.(0.28ml, 2.7mmol), (1.06 restrain, 2.1mmol) to add Michael-adduct then to add pyrrolidine in this solution.This mixture was reduced to room temperature in 1 hour, shook then 20 hours.In argon, in reflux, this reaction was finished in this reactant mixture heating in 1 hour afterwards.Solvent evaporated is with anxious poly-chromatography (SiO ₂, eluent: hexane-EtOAC, 9: 1) and the purification residue, obtain Boc-Tbg-CH ₂-(RS)-CH (CH ₂C (O) CMe ₃) C (O) OEt (0.77 gram, 83%).After this, (0.77 gram 1.7mmol) is dissolved in glycol dimethyl ether-H to a back chemical compound that obtains ₂O (1: 1,10ml) in.(0.31 gram 7.4mmol) joins in this solution with a hydronium(ion) oxidation lithium.This mixture is at room temperature vibrated 4 hours, make it to be acid, and (3 * 25ml) extract with EtOAc with 10% aqueous citric acid solution (20ml).With extract drying (MgSO ₄) and under reduced pressure, concentrate, obtaining the title compound of present embodiment, (0.69 gram is from Boc-Tbg-CH for a kind of brown solid ₂C (O) O-CH ₂CH=CH ₂Gross production rate be 80%).This product is that a kind of ratio is the mixture (being proved by NMR) of 55: 45 diastereomer. ¹H NMR (CDCl ₃) the result be δ 0.97 (s, 9H, less important isomer), 0.99 (s, 9H, main isomer), 1.14 (s, 18H), 1.48 (s, 18H), and 2.68-3.12 (m, 8H), 3.30 (m, 2H), 4.08 (d, J=9Hz, 1H, less important isomer), 4.10 (d, J=9Hz, 1H, main isomer), 5.11 (d, J=9Hz, 2H).

Embodiment 4

Intermediate product H-Tbg-CH ₂CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn) (Bzl)-NH-(S)-CH (CH ₂C (O) Me ₃) CH ₂The preparation of OBzl

Coupling step according to embodiment 1, utilize embodiment 2 title compound (3.42 the gram, 7.13mmol) as first reactant, utilize embodiment 3 title compound (2.50 the gram, 6.48mmol) as second reactant, the coarse products thing is carried out the poly-chromatography (SiO of urgency ₂, eluant: hexane-EtOAc, 4: 1) obtain corresponding title compound the N-Boc derivant (4.64 the gram, 84%; Rf=0.21, hexane-EtOAc, 7: 3).(4.64 grams, 5.44mmol) solution that is dissolved in the 6N HCl/ diox (50ml) at room temperature vibrates 1 hour, concentrates under reduced pressure then with this derivant.Residue is dissolved in Et ₂Among the O.With saturated NaHCO ₃The solution that aqueous solution and salt water washing obtain is with its drying (MgSO ₄), and concentrate and to obtain the yellow oil formed by two kinds of diastereomers.Adopt anxious poly-chromatography (SiO ₂, eluent: hexane-EtOAc-MeOH, 5: 4.5: 0.5) and separate this two kinds of isomers.Obtain required isomer (the i.e. more polarization (themore polar) of colorless oil form (2.52 grams, 52%); Rf=0.18, EtOAc-hexane-MeOH, 7: 3: 0,5).Directly use this isomer below among the embodiment: the title compound of present embodiment, and be not further purified.

Embodiment 5

Boc-(N-Me) Val-Tbg-CH ₂(R)-CH (CH ₂C (O) CMe ₃) C (O)-ASP (cyPn) (Bzl)-NH-(S)-CH (CH ₂CMe ₃) CH ₂The preparation of OBzl

Coupling step method according to embodiment 1, utilize title compound (4.45 grams of embodiment 4,5.95mmol) (4.41 grams 17.9mmol) as second reactant, carry out the poly-chromatography analysis (SiO of urgency with raw product as first reactant and with the N-methylvaline ₂, eluent: hexane-EtOAc, 7: 3) and obtain the title compound (4.02 gram, 72% productive rate) of present embodiment, ¹H NMR (CDCl ₃) δ 0.87 (d, J=7Hz, 6H), 0.90 (s, 18H), 1.10 (s, 9H), 1.48 (s, 9H), 1.5-2.0 (m, 10H), 2.30 (m, 1H), 2.5-3.1 (m, 5H), 2.80 (s, 3H), 3.30 (m, 2H), 4.0 (d, J=12.5Hz, 1H), 4.20 (m, 1H), 4.32 (d, J=8.5Hz, 1H), 4.49 (d, J=4.5Hz, 2H), 4.64 (d, J=11Hz, 1H), 5.18 (d, J=5Hz, 2H), 6.78 (wide d, J=8.5Hz, 1H), 7.11 (wide d, J=8.5Hz, 1H), 7.18 (wide d, J=11Hz, 1H), 7.2-7.45 (m, 10H).

Embodiment 6

PhCH ₂CH ₂C (O)-(N-Me) Val-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) preparation of C (O)-Asp (cyPh)-γ MeLeucinol

Title compound (4.02 grams with embodiment 5,4.25mmol) be dissolved in the solution that obtains in the 6N HCl/ diox (30ml) and at room temperature vibrated 1 hour, under reduced pressure concentrate the derivant of the free N-terminal amino group of tool of the title compound of the embodiment 5 that obtains its hydrochloride form then.After this, according to the coupling step of embodiment 1, and with this aminoderivative (2.00 grams 13.3mmol) as second reactant, adopt anxious poly-chromatography (SiO as first reactant and with benzenpropanoic acid ₂, eluent: hexane-EtOAc, 3: 2) and thick purifying products is obtained PhCH ₂CH ₂C (O)-N-Me-Val-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp-(cyPn) (OBzl)-NH-(S)-CH (CH ₂CMe ₃) CH ₂OBzl, a kind of white foam agent form (4.00 grams, 94%; Rf=0.35, hexane-EtOAc, 1: 1).

With chemical compound (4.00 grams, the 4.03mmol) hydrogenolysis { 20%Pd (OH) that obtains ₂/ C (200mg), 1 atmospheric H ₂, EtOH, 5 hours }.After this reaction is finished, by catalyst being removed from reactant mixture with 45 μ m membrane filtrations.Under reduced pressure, filtrate concentrating obtained a kind of clean oil.This oil is dissolved in Et ₂O (100ml).Under reduced pressure, this solution evaporation is extremely done.To dissolve and evaporation step repeat, until obtaining a kind of white solid.Grind this solid with hexane, filter, and drying obtains title compound (3.12 grams, 95%) under reduced pressure.Its ¹H NMR (d ₆-DMSO) be 400 MHz; Annotate: this chemical compound is that 50: 50 form of mixtures is present among the DMSO with two kinds of rotation foreign body object ratios, and its δ 0.71-0.92 (m, 24H), 1.05 (s, 4.5H), 1.06 (s, 4.5H), 1.20-1.78 (m, 10H), and 1.93-2.16 (m, 2H), 2.48-2.83 (m, 6H), 2.84 (s, 1.5H), 2.92 (s, 1.5H), 2.96-3.06 (m, 1H), 3.10-3.23 (m, 2H), and 3.72-3.81 (m, 1H), 4.09-4.14 (m, 1H), 4.22 (d, 8Hz, 0.5H), 4.54-4.62 (wide m, 1H), 4.73-4.81 (m, 1.5H), 7.12-7.29 (m, 6H), 7.94 (d, J=10Hz, 1H), 8.04 (d, J=8Hz, 0.5H), 8.32 (d, J=8.5Hz, 0.5H).

According to the step of embodiment 6, but replace benzenpropanoic acid, obtain (PhCH with 2-(benzyl)-3-benzenpropanoic acid (dibenzyl acetic acid) ₂) ₂CHC (O)-(N-Me) Val-Tbg-CH ₂-(R)-CH (CH ₂C (O)-CMe ₃) C (O)-Asp (cyPh)-γ MeLeucinol.

Embodiment 7

Preparation Et ₂CHNHC (O)-Tbg-CH ₂-R-(CH ₂C (O)-CMe ₃) CO-Asp (cyPn)-γ MeLeucinol

With Carbimide. 1-ethyl propyl ester (28mg, 0.248mmol) be added to embodiment 4 title compound (23mg, 0.030mmol) and triethylamine (6mg 0.057mmol) is dissolved in anhydrous CH ₂Cl ₂In in the solution that obtains.At argon, with this reactant mixture vibration 1 hour, at room temperature vibrate 18 hours then under 0 ℃.TLC (EtOAc-hexane, 1: 1) indicates finishing of this reaction.Under reduced pressure, solvent evaporation is fallen.Adopt anxious poly-chromatography (SiO ₂, eluent: hexane-EtOAc, 6: 4) and residue purified obtained the corresponding dibenzyl derivant (16mg) of present embodiment title compound. ¹H?NMR(400?MHz，CDCl ₃)δ0.9(t，J＝7Hz，3H)，0.92(t，J＝7Hz，3H)，0.94(s，9H)，0.95(s，9H)，1.10(s，9H)，1.25-1.90(m，14H)，2.52(m，1H)，2.68(m，1H)，2.81(m，1H)，2.94-3.07(m，2H)，3.27(dd，J＝7.2，9Hz，1H)，3.36(dd，J＝5.5，9Hz，1H)，3.46(m，1H)，4.07(d，J＝9Hz，1H)，4.23(m，1H)，4.28(d，J＝9Hz，1H)，4.48(dd，J＝10Hz，2H)，4.66(d，J＝10Hz，1H)，4.74(d，J＝9Hz，1H)，5.17(dd，J＝14Hz，2H)，7.10(d，J＝8.5Hz，1H)，7.2-7.45(m，11H)。

(10% is carried on the palladium on the carbon, and 1 atmospheric pressure EtOH) obtains this motif compound with the dibenzyl derivant hydrogenolysis that obtains.Mass spectrum: 703 (M+Na) ⁺

Embodiment 8

Be used to make the preparation of other representative intermediate of C-end of the peptide of structural formula 1

(a) NH ₂-(R)-CH (Et) CMe ₃: to 4, and 4-dimethyl-propione (106 grams, 0.92mmol) and (R)-(111 grams 0.92mmol) are dissolved in that (0 ℃) adds TiCl in the cooling solution that obtains in the benzene (1 liter) to α-xylylamine ₄(50.5ml 0.46mmol) is dissolved in solution in the benzene (200ml), and the speed of interpolation should keep the temperature of this mixture to be lower than below 10 °.After this, with this mixture 40 ℃ of mechanical vibration 3 hours, cool to room temperature, and use diatomite filtration.Use Et ₂O washs kieselguhr.The filtrate and the cleaning mixture that merge are concentrated.Residue is dissolved among the anhydrous MeOH (2 liters).This solution is cooled to 0 ℃, when mixture temperature being maintained at below 5 ℃ with NaBH ₄(20 restrain, the adding of 0.53mmol) merotomizing.Methanol is evaporated.Residue is dissolved in Et ₂Among the O.With this solution of salt water washing, dry (MgSO ₄) and be evaporated to driedly, obtain a kind of reddish oil (this is that a kind of ratio is the mixture of 18: 1 diastereomer to differentiate indication by NMR).Should the anxious poly-chromatography (SiO of oil ₂, eluent: EtOAc/ hexane, 7: 93) and purification obtains a kind of N-(1 (R)-phenethyl)-1 (R)-ethyl-2 of liquid form, 2-dimethyl propylamine (110 grams, 54%).This material is dissolved in the hexane (1.5 liters).To be dissolved in 6N HCl in the diox (90ml) adds in this solution in during 15 minutes.Collect the white solid that obtains with filter paper, the washing of Yong diox obtains N-(1 (R)-phenethyl)-1 (R)-ethyl-2 then, 2-dimethyl propyl hydrochloride (125 grams, 97%). ¹H?NMR(CDCl ₃)δ0.55(t，J＝7.5Hz，3H)，1.14(s，9H)，1，54-1.95(m，2H)，2.23(d，J＝6.5Hz，3H)，2.36-2.44(m，1H)，4.31-4.49(m，1H)，7.30-7.48(m，3H)，7.74-7.79(m，2H)。

The chemical compound (41.5 gram) that obtains is dissolved in solution and 10% (w/w) Pd/C that obtains among the MeOH (120ml) mixes, at room temperature,, and vibrate this mixture 48 hours under the 50psi hydrogen condition at a Parr hydrogenator.This mixture is filtered, filtrate is concentrated into a kind of required NH of white solid form ₂-(R)-CH (Et) CMe ₃, its salt acid-addition salts form (25g, 100%). ¹H NMR (CDCl ₃) δ 1.10 (s, 9H), 1.22 (t, J=7Hz, 2H), 1.58-1.90 (m, 2H), 2.70-2.85 (m, 1H), 8.10-8.40 (wide s, 3H).

With same method, but in front the step with 3,3-dimethyl-2-butanone replacement 4,4-dimethyl-propione obtains NH ₂-(R)-CH (Me) CMe ₃HCl.

Embodiment 9

Other the representative intermediate of N-end that are used to make up the peptide derivant of structural formula 1 according to the preparation of the step of embodiment 7

(a) Carbimide. 1-propyl group butyl ester: adopt V.S.Goldesmidt and M.Wick, Lie-bigs Ann.Chem., 575,217 (1952) method prepares this intermediate from the amino heptane of the 4-that buys.

(b) Carbimide. 1-ethyl-1-(2-acrylic)-3-butene esters:

(14.5 grams 264mmol) are dissolved in anhydrous Et with propionitrile ₂The drips of solution that O (40ml) obtains is added to 1.0M pi-allyl bromination magnesium/Et ₂Among the O (880ml).With the mechanical vibration 2 hours in reflux of this reactant mixture.After this it is cooled to 0 ℃.With NH ₄Cl saturated aqueous solution (320ml) is added in this reaction mixture carefully.Separate organic facies, dry (MgSO ₄), be chilled to 0 ℃, then under uniform temp with 1M HCl/Et ₂O (200ml) mixes.The solid that collection obtains, drying under reduced pressure (about 27 grams).The material that obtains is dissolved in CH ₂Cl ₂(200ml).Na with 10% (w/v) ₂CO ₃This solution of (2 *) solution washing, then with the salt washing, dry (MgSO ₄) and be concentrated into driedly, obtain a kind of yellow oil.Should oil distillation (82-85 °/20 Torr) obtain 1-ethyl-1-(2-acrylic)-3-butenylamine (11.6 grams, 34%), it is a kind of colourless liquid; ¹H NMR (CDCl ₃), 400MHz) δ 0.89 (t, J=7Hz, 3H), 1.39 (q, J=7Hz, 2H), 2.11 (d, J=7Hz, 2H), 5.06-5.14 (m, 4H), 5.80-5.89 (m, 2H).

Adopt the V.S.Goldesmidt of above description and the method for M.Wick that the chemical compound that obtain above is converted into Carbimide. 1-ethyl-1-(2-acrylic)-3-butene esters.

First step of front b joint method promptly prepares 1-ethyl-1-(2-acrylic)-3-butenylamine, is based on by G.Alvernhe and A.Laurent TetrahedronLett., 1057 (1973) integrated approach of describing.Utilize this integrated approach, and selective response thing suitably, can prepare other essential isocyanates intermediate, these intermediate are applicable to final preparation at the terminal unsaturated peptide derivant of N-, and promptly a kind of unsaturated alkyl amino-carbonyl is 1-methyl isophthalic acid-(2-acrylic)-3-cyclobutenyl for example.But it should be noted that so last product will be the corresponding peptide derivant of structural formula 1 when essential isocyanates intermediate is used for the scheme of embodiment 7, wherein the N-end is saturated.And this last step is a kind of such corresponding peptides for preparing; For example, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, (FAB/ mass spectrum (m/z): 693[M+H] ⁺, the subject peptide derivant of indicating among the embodiment 10 hereinafter) effective ways.

Therefore, use suitable intermediate, use a series of coupling steps of embodiment 1-7 and go to protect step, can prepare other chemical compound of structural formula 1, for example those that enumerate in the table of embodiment below.In some cases, the precipitation of end-product does not provide purifying substance, so just can utilize the acetonitrile and the water gradient that respectively contain 0.06%TFA to carry out half preparation HPLC analysis with this purifying products on the C-18 reversed-phase column.In order to finish this process, crude product is dissolved in 0.1M NH ₄Recall to about 7 with the pH of 0.1M AcOH aqueous solution in the OH aqueous solution and before purification this solution.If be suitable for, available this mode is separated non-enantiomer mixture.

Several examples of other chemical compounds of the structural formula 1 that can prepare are (PhCH ₂) ₂CHC (O)-(N-Me) Val-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃(PhCH ₂) ₂CHC (O)-(N-Me) Val-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Me) CMe ₃

Other several examples of the chemical compound of structural formula 1 are: Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃, FAB/ mass spectrum (m/z): 665[M+H] ⁺EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, FAB/ mass spectrum (m/z): 722[M+H] ⁺And EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃, FAB/ mass spectrum (m/z): 693[M+H] ⁺

Embodiment 10

The show peptide derivant, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O)-Me ₃)-C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, and the conjugate of this derivant and acycloguanosine resists a kind of wild type HSV-1 clinical separation strain, and the Research on effect of two kinds of acycloguanosine resistance HSV-1 clinical separation strains.

Virus: these clinical separation strains are by people such as S.L.Sacks, Annals of Inte-rnal Medicine, and 111,893 (1989) describe.They come from 25 a years old women who suffers from acute myelogenous leukemia and recur for the first time behind allogeneic bone marrow transplantation.Transplant the starting culture of finding herpes simplex virus on the 8th day in the back be positive (294 separated strains are defined as wild type HSV-1) at bone.Begin then with the acycloguanosine treatment, this viral cultures still was positive (615 separated strains are represented the HSV-1 resistant strain) in the 36th day.To a kind of inferior separated strain 615.9 (ACV ^IThe TK clinical separation strain) carries out specificity analysis and find it, but phosphine formic acid sensitivity and its thymidine kinase (TK) activity are also weakened acycloguanosine tool resistance.Confirm that subsequently 615.9 is a kind of TK deficiency mutants.On the contrary, 615.8 inferior separated strain (ACV ^IThe POL clinical separation strain) characteristic is that acycloguanosine and phosphine formic acid tool resistance and its thymidine kinase activity are approached observed activity in 294 separated strains before treatment.Confirm that subsequently 615.8 is a kind of dna polymerase mutant bodies.

Analyze:

At 150cm ²T-flask (1.5 * 10 ⁶Individual cell/bottle) in BHK-21/C13 cell (ATCC CCL 10) was cultivated 2 days, α-MEM the culture medium (Gibco Canada Inc., the Burlington that have replenished 8% (v/v) hyclone (FBS, Gibco Canada Inc.) are housed in this culture bottle, Ontario, Canada).With the cell trypsinization, transfer to then in the fresh culture of a flat board that 24 holes are arranged, make and contain 2.5 * 10 in the 750 μ l culture medium in each hole ⁵Individual cell.Cell was cultivated 6 hours at 37 ℃, so that make cell adhesion to this flat board.After this, with the 500 μ l α-MEM that replenished 0.5% (v/v) FBS (hyclone) once, use the same medium (low serum) of 750 μ l to cultivate then 3 days with cell washing.Standing this serum starvation after the stage, removing this low blood serum medium and cell was cultivated 2-3 hour in the BBMT of 500 μ l.[the BBMT culture medium is by people such as P.Braz-eau, Proc.Natl.Acacl.Sci.USA, 79,7909 (1982) describe].After this, in the BBMT culture medium of 100 μ l, use HSV-2 infection cell (infection multiplicity=0.02PFU/ cell).(note: the HSV-2 of use is the HG-52 strain, referring to Y.Langelierand G.Buttin, and J.Gen.Virol., 57,21 (1981); This virus is stored in-80 ℃).After 37 ℃ of viruses absorb 1 hour, remove culture medium and use BBMT washed cell (3 * 250 μ l).With the cell in each hole with or the detectable that is dissolved in the 200 μ l BBMT culture medium that need not (contrast) debita spissitudo cultivate together.After 37 ℃ are cultivated 29 hours,,, thereby collect infected cell subsequently with the cell freeze thawing at first at-80 ℃ of frozen cells.Under the Borneolum Syntheticum that melts helps, the cell in each hole is scraped off from hole surface.After complete freeze thawing, the collecting cell suspension also embathes each hole with the culture medium of 150 μ l BBMT.At 4 ℃ with light and slow ground of this virus sample (suspension+washing liquid) supersound process 4 minutes.Remove cell debris by centrifugal (at 4 ℃ with centrifugal 10 minutes of 1000 * g).Collect supernatant, and be stored in-80 ℃, up to being used to measure virus titer.

With people such as M.Langlois, Journal of Biological Standardizati-on is used to measure virus titer after 14,201 (1986) the colorimetric analysis correct.

More particularly, in the same manner of Miao Shuing, with trypsin treatment BHK-21/C13 cell, and transfer in the fresh culture of one 96 hole microdroplet flat board, in the above so that contain 20,000 cells in the 100 μ l culture medium in each hole.To cultivate 2 hours at 37 ℃ at the cell in the flat board that this prepares.During this period, with the virus sample freeze thawing, then with its supersound process 15 seconds slightly, and prepare the grade diluent (1/5 dilution factor: the sample of 50 μ l adds the BBMT culture medium of 200 μ l, finishes the serial dilution process with a multichannel pipet) of this sample.

Recited above the BHK-21/C13 cell culture was finished in 2 hours after, substitute wherein culture medium with the α-MEM culture medium of having replenished 3% (V/V) FBS.At this moment cell has been ready to accept the infection of various virus sample diluent.Various dilution aliquot sample (50 μ l) are transferred in the dull and stereotyped appropriate bore.The infected cell that obtains was cultivated 2 days at 37 ℃.0.15% (V/V) dimethyl diaminophenazine chloride dye solution, the 50 μ l that will be dissolved in then in the Hank balanced salt solution (pH 7.3, Gibco Canada Inc.) are added in each hole.This flat board for preparing was cultivated 45 minutes at 37 ℃.Sucking-off culture medium from each hole then, with the Hank balanced salt solution of 200 μ l once with cell washing.After washing, add 0.1MSorensen citrate buffer (pH 4.2) and alcoholic acid 1: 1 mixture 100 μ l, dyestuff is discharged from cell.[the Sorensen citrate buffer prepares as follows: at first, the filterable water of mistake that is dissolved in citric acid monohydrate compound (21 gram) in the 1N NaOH aqueous solution (200ml) and adds q.s reaches 1 liter of volume, thereby has prepared 0.1M disodium citrate solution.Second step, 0.1M disodium citrate solution (61.2ml) is mixed with 0.1NHCl aqueous solution (38.8ml), if necessary, the pH of the solution that obtains is adjusted to 4.2].Mixture in the hole is accepted light and slow vibration, mix rightly guaranteeing.With a spectrophotometer plate reader hole on the flat board is scanned at the 540nm place, estimated the survivaling cell number.The test agent that detects various concentration by this way suppresses percentage rate to viral growth, and can calculate that to cause the concentration of 50% the downtrod test agent of virus replication be IC ₅₀

Aforementioned analytical method is used for carrying out independent and their various compositionss are estimated to acycloguanosine and peptide derivant.Thus, can prove the comparative effectiveness of the anti-strain of preventing or cure a disease of two kinds of chemical compounds.Moreover, the result who obtains in these researchs used the method assessment of isobole and confirm the synergy of two kinds of chemical compounds; Referring to J.Suhnel, J.Antivir-al Research, 13,23 (1990) the isobole methods of describing.

To the clearer description of isobole method, produce when this method need be used separately by two kinds of test compounds, and have the experimental data that equivalent answers the conjugate of various dose of two kinds of chemical compounds of level to produce.According to this method, add the subject peptide derivant (IC of selected concentration ₅, IC ₁₀, IC ₂₀And IC ₃₀) with the compositions of the acycloguanosine of various dose, estimation IC ₅₀In this experiment, the IC of subject peptide derivant ₅, IC ₂₀And IC ₃₀And the dosage of acycloguanosine all comes from the curve that obtains previously.Utilization is called FIC ₆₀The value of (acycloguanosine) ratio of required concentration (this value be the concentration that suppresses the required acycloguanosine of 60% HSV under the situation that the subject peptide derivant of fixed concentration exists when this peptide derivant does not exist) produces isobologram (isobologram).The value of the ratio of the concentration of the peptide derivant that it is duplicated with respect to the fixed concentration of representative peptide derivant and the HSV that reduces by 60% when not having acycloguanosine is drawn collection of illustrative plates.Equation:

X takes out:

Y-axis:

The result:

Table 1 has only been enumerated in the described in front cell culture detection method of carrying out with the HSV-1 clinical separation strain of wild type HSV-1 clinical separation strain and two kinds of tool acycloguanosine resistances the result who obtains when subject peptide with acycloguanosine and present embodiment detects.

Table 1

Separated strain separated strain separated strain separated strain chemical compound 294 615.8 615.9

IC ₅₀(μ M) IC ₅₀(μ M) IC ₅₀(μ M) acycloguanosine 1.1 19 55 subject peptide derivants 2.0 2.0 6.2

The described result of table 1 has proved that structural formula 1 described chemical compound has the activity of anti-wild type HSV-1, and these chemical compounds are to the HSV-1 of tool acycloguanosine resistance, and for example POL mutant and TK deficiency mutant have same opposing effect; And its efficient reduces a lot (than the low 20-40 of above-mentioned efficient doubly) when resisting these mutants with acycloguanosine.

Following Table II, III and IV represent according to wild type HSV-1 clinical separation strain and two kinds of cell culture detection methods noted earlier that tool acycloguanosine resistance HSV-1 clinical separation strain carries out, the result who obtains during the compositions of the subject peptide derivant of estimation acycloguanosine and present embodiment.

Table II

Antiviral activity

Synergistic research

Separated strain 294

The IC of acycloguanosine during fixed concentration that (treatment before clinical separation strain) subject peptide derivant and subject peptide derivant make up ₅₀ ⁽¹⁾(μ M) (μ M) 0.3 0.85 0.5 0.70 0.7 0.35 0.9 0.30 1.0 0.30 1.2 0.22 1.4 0.10 1.6 0.13

Table III

Antiviral activity

Synergistic research

Separated strain 615.8

(ACV ^IPOL treats separated strain) IC of acycloguanosine during the fixed concentration of subject peptide derivant and subject peptide derivant combination ₅₀ ⁽²⁾(μ M), (μ M) 0.3 15.0 0.5 13.0 0.7 7.0 0.9 8.0 1.0 5.3 1.2 5.0 1.4 1.7 1.6 2.4

Table IV

Antiviral activity

Synergistic research

Separated strain 615.9

(ACV ^ITK treats separated strain) IC of acycloguanosine during the fixed concentration of subject peptide derivant and subject peptide derivant combination ₅₀ ⁽²⁾(μ M) (μ M) 1.0 33.0 2.0 17.0 3.0 5.5 4.0 5.6 5.0 2.0 6.0 0.9 7.0＜0.02 8.0＜0.02

About the note in four tables in front:

IC ₅₀Value is not proofreaied and correct at dissolubility.

All stock solution were all carried out super centrifugal 30 minutes with 100,000 * g at 4 ℃ before using.

The dissolubility of this title peptide derivant when 20 μ M is 85%.

The cytotoxicity of this title peptide derivant is 89 μ M (this value is proofreaied and correct).

(1) the subject peptide derivant of the corresponding fixed concentration that shows in the on the left side hurdle exists down, when the concentration of acycloguanosine is 7.6 * 10/ ^-4To 1.5 * 10 ¹Obtain IC on the dose response curve that produces in the time of in the scope ₅₀

(2) the title peptide derivant of the corresponding fixed concentration that shows in the on the left side hurdle exists down, when the concentration of acycloguanosine is 1.5 * 10 ^-4To 1.5 * 10 ²The IC that obtains on the dose response curve that produces in the time of in the scope ₅₀

Table II, III and IV show that when two kinds of reagent were used in combination, the peptide of structural formula 1 can be strengthened the activity of the HSV-1 mutant of acycloguanosine opposing wild type HSV-1 and tool acycloguanosine resistance.This result shows when peptide and the increase of acycloguanosine concentration ratio, the IC of acycloguanosine ₅₀Proportional reduction.

The positive findings (Synergistic) that accompanying drawing 1 obtains when being the research of graphic representation application isobole method, this research is the title peptide with acycloguanosine and present embodiment, and inferior separated strain 615.8 (ACV ^γThe POL clinical separation strain) carries out.

Equally, accompanying drawing 2 is graphic representation title peptides at usefulness acycloguanosine and present embodiment, and inferior separated strain 615.9 (ACV ^γTK) positive findings that obtains in the research of carrying out.

Claims (25)

1, a kind of peptide derivant is to the application in the herpes simplex viral infections of mammal treatment tool acycloguanosine resistance, and wherein, described peptide derivant is an acceptable salt on the chemical compound of structural formula 1 expression or its therapeutics

A-B-D-CH ₂CH{CH ₂C(O)R ¹}C(O)-NHCH{CR ²(R ³)COOH}C(O)-E

1 wherein A be phenylacetyl group, hydrocinnamoyl, (4-aminophenyl) propiono, (4-fluorophenyl) propiono, (4-hydroxy phenyl) propiono, (4-methoxyphenyl) propiono, 2-(benzyl)-3-hydrocinnamoyl, 2-{ (4-fluorophenyl) methyl }-3-(4-fluorophenyl) propiono, 2-{ (4-methoxyphenyl) methyl }-3-(4-methoxyphenyl) propiono or benzylamino carbonyl; B is (N-Me)-Val or (N-Me)-Ile; Or A and B form a kind of saturated alkyl amino-carbonyl together, it is selected from following groups: the butyl amino carbonyl, 1-Methylethyl amino carbonyl, 1-methyl-propyl amino carbonyl, 1-ethyl propyl amino carbonyl, 1,1-dimethyl ethyl butyl amino carbonyl, 1-ethyl-butyl amino carbonyl, 1-propyl group butyl amino carbonyl, 1-ethyl pentyl group amino carbonyl, 1-butyl amyl group amino carbonyl, 1-ethyl-butyl amino carbonyl, 2-ethyl pentyl group amino carbonyl, 1-methyl isophthalic acid-propyl group butyl amino carbonyl, 1-ethyl-1-propyl group butyl amino carbonyl, 1,1-dipropyl butyl amino carbonyl, (1-propyl group cyclopenta) amino carbonyl and (1-propyl group cyclohexyl) amino carbonyl; D is Val, Ile or Tbg; R ¹Be the 1-Methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 1,1-dimethyl propyl, 2,2-dimethyl propyl, cyclobutyl, cyclopenta, cyclohexyl, 1-methylcyclopentyl, NR ⁴R ⁵R wherein ⁴Be hydrogen or low alkyl group, and R ⁵Be low alkyl group, perhaps R ⁴And R ⁵Form pyrrolidinyl with its bonded nitrogen-atoms, piperidino, morpholino or 4-methyl isophthalic acid-piperazinyl; R ²Be hydrogen, R ³Be methyl, ethyl, the 1-Methylethyl, 1, the 1-dimethyl ethyl, propyl group, 2-acrylic or benzyl carry R ²And R ³Carbon atom have (R)-configuration, perhaps R ²And R ³Be separately methyl or ethyl, perhaps R separately ²And R ³Form cyclobutyl with its bonded carbon atom, cyclopenta or cyclohexyl; And E is NHR ⁶R wherein ⁶Be the 2-methyl-propyl, 2,2-dimethyl propyl, 1 (R), 2,2-trimethyl propyl group, 1,1,2,2-tetramethyl propyl group, 1 (R)-ethyl-2,2-dimethyl propyl, 2-(R, S)-methyl butyl, 2, the 2-dimethylbutyl, 3,3-dimethylbutyl, 1 (R), 2,2-trimethyl butyl, 1 (R), 3,3-trimethyl butyl, the 2-ethyl-butyl, 2,2-diethyl butyl, 2-ethyl-1 (R)-methyl butyl, 2-ethyl-2-methyl butyl, 1 (R)-ethyl-3, the 3-dimethylbutyl, 2,2-dimethyl amyl group, cis or trans-2-methylcyclohexyl, 2,2-Dimethylcyclohexyl or cyclohexyl methyl; Perhaps E is NHCH (R ⁷)-Z wherein carries R ⁷Carbon atom have (S)-configuration, R ⁷Be 1, the 1-dimethyl ethyl, the 1-methyl-propyl, the 2-methyl-propyl, 2,2-dimethyl propyl or cyclohexyl methyl, and Z is CH ₂OH, C (O) OH, C (O) NH ₂Or C (O) OR ⁸R wherein ⁸Be methyl, ethyl or propyl group.

2, the application of peptide derivant according to claim 1, wherein, this peptide derivant is an acceptable salt on the chemical compound of structural formula 1 or its therapeutics, wherein A is a hydrocinnamoyl, 2-(benzyl)-3-hydrocinnamoyl or benzylamino carbonyl; B is (N-Me) Val; D is Tbg; R ¹Be the 1-Methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 1,1-dimethyl propyl, 2,2-dimethyl propyl, cyclobutyl, cyclopenta, cyclohexyl or 1-methylcyclopentyl; R ²Be hydrogen, R ³Be methyl, ethyl, the 1-Methylethyl, propyl group or benzyl, and carry R ²And R ³Carbon atom have (R) configuration, perhaps R ²And R ³Be separately methyl or ethyl, perhaps R separately ²And R ³Form cyclobutyl with its bonded carbon atom, cyclopenta or cyclohexyl; E is NHR ⁶, R wherein ⁶Be 2, the 2-dimethyl propyl, 1 (R), 2,2-trimethyl propyl group, 1 (R)-ethyl-2, the 2-dimethyl propyl, 2,2 one dimethylbutyls or 1 (R)-ethyl-3,3-dimethylbutyl or E are NHCH (R ⁷The carbon atom that)-Z wherein carries R7 has (S) configuration, R ⁷Be 2,2-dimethyl propyl, Z are CH ₂OH, C (O) OH, C (O) NH ₂Or C (O) OR ⁸, R wherein ⁸Be methyl, ethyl or propyl group.

3, the application of peptide derivant according to claim 1, wherein this peptide derivant is an acceptable salt on structural formula 1 described chemical compound or its therapeutics, wherein A and B form a kind of saturated alkyl amino-carbonyl together, it is selected from down group: 1-ethyl propyl amino carbonyl, 1-ethyl-butyl amino carbonyl, 1-propyl group butyl amino carbonyl, 2-ethyl pentyl group amino carbonyl, 1-methyl isophthalic acid-propyl group butyl amino carbonyl, 1-ethyl-1-propyl group butyl amino carbonyl, 1,1-dipropyl butyl amino carbonyl and (1-propyl group cyclopenta) amino carbonyl; D is Tbg; R ¹Be the 1-Methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 1,1-dimethyl propyl, 2,2-dimethyl propyl, cyclobutyl, cyclopenta, cyclohexyl or 1-methylcyclopentyl; R ²Be hydrogen and R ³Be methyl, ethyl, the 1-Methylethyl, propyl group or benzyl carry R ²And R ³Carbon atom have (R)-configuration, perhaps R ²And R ³Be separately methyl or ethyl, perhaps R separately ²And R ³Carbon atom bonded-work forming cyclobutyl, cyclopenta or cyclohexyl with it; E is NHR ⁶R wherein ⁶Be 2, the 2-dimethyl propyl, 1 (R), 2,2-trimethyl propyl group, 1 (R)-ethyl-2, the 2-dimethyl propyl, 2,2 one dimethylbutyls or 1 (R)-ethyl-3,3-dimethylbutyl or E are NHCH (R ⁷)-Z wherein carries R ⁷Carbon atom have (S) configuration, R ⁷Be 2,2-dimethyl propyl, Z are CH ₂OH, C (O) OH, C (O) NH ₂Perhaps C (O) OR ⁸, R wherein ⁸Be methyl, ethyl or propyl group.

4, the application of the described peptide derivant of claim 1, wherein this peptide derivant is selected from down group: PhCH ₂CH ₂C (O)-(N-Me) Val-Tbg-CH ₂-(R)-CH (CH ₂C (O)-CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Et ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃, EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, and EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃.

5, the described peptide derivant of claim 1, or acceptable salt and a kind of antiviral nucleoside analogue on its therapeutics, or the application of compositions in the herpes simplex viral infections of the mammiferous tool acycloguanosine resistance of treatment of acceptable salt on its therapeutics.

6, the application of compositions according to claim 5, wherein this peptide derivant is an acceptable salt on a kind of chemical compound of structural formula 1 or its therapeutics, wherein A is a hydrocinnamoyl, 2-(benzyl)-3-hydrocinnamoyl or benzylamino carbonyl; B is (N-Me) Val; D is Tbg; R ¹Be the 1-Methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 1,1-dimethyl propyl, 2,2-dimethyl propyl, cyclobutyl, cyclopenta, cyclohexyl or 1-methylcyclopentyl; R ²Be hydrogen and R ³Be methyl, ethyl, the 1-Methylethyl, propyl group or benzyl carry R ²And R ³Carbon atom have (R)-configuration, perhaps R ²And R ³Be separately methyl or ethyl, perhaps R separately ²And R ³Form cyclobutyl with its bonded carbon atom, cyclopenta or cyclohexyl; And E is NHR ⁶R wherein ⁶Be 2, the 2-dimethyl propyl, 1 (R), 2,2-trimethyl propyl group, 1 (R)-ethyl-2, the 2-dimethyl propyl, 2,2-dimethylbutyl or 1 (R)-ethyl-3,3-dimethylbutyl or E are NHCH (R ⁷)-Z wherein carries R ⁷Carbon atom have (S) configuration, R ⁷Be 2,2-dimethyl propyl, Z are CH:OH, C (O) OH, C (O) NH: perhaps C (O) OR ⁸R wherein ⁸Be methyl, ethyl or propyl group.

7, according to the application of the described compositions of claim 5, wherein this peptide derivant is an acceptable salt on the chemical compound of structural formula 1 or its therapeutics, A and B form a kind of saturated alkyl amino-carbonyl together in this structural formula 1, it is selected from down group: 1-ethyl propyl amino carbonyl, 1-ethyl-butyl amino carbonyl, 1-propyl group butyl amino carbonyl, 2-ethyl pentyl group amino carbonyl, 1-methyl isophthalic acid-propyl group butyl amino carbonyl, 1-ethyl-1-propyl group amino carbonyl, 1,1-dipropyl butyl amino carbonyl and (1-propyl group cyclopenta) amino carbonyl; D is Tbg; R ¹Be the 1-Methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 1,1-dimethyl propyl, 2,2-dimethyl propyl, cyclobutyl, cyclopenta, cyclohexyl or 1-methylcyclopentyl; R ²Be hydrogen and R ³Be methyl, ethyl, the 1-Methylethyl, propyl group or benzyl, and carry R ²And R ³Carbon atom have (R)-configuration, perhaps R ²And R ³Be separately methyl or ethyl, perhaps R separately ²And R ³Form cyclobutyl with its bonded carbon atom, cyclopenta or cyclohexyl; E is NHR ⁶R wherein ⁶Be 2, the 2-dimethyl propyl, 1 (R), 2,2-trimethyl propyl group, 1 (R)-ethyl-2, the 2-dimethyl propyl, 2,2-dimethylbutyl or 1 (R)-ethyl-3,3-dimethylbutyl or E are NHCH (R ⁷)-Z wherein carries R ⁷Carbon atom have (S) configuration, R ⁷Be 2,2-dimethyl propyl, Z are CH ₂OH, C (O) OH, C (O) NH ₂Perhaps C (O) OR ⁸, R wherein ⁸Be methyl, ethyl or propyl group.

8, according to the application of the described compositions of claim 5, wherein this peptide derivant is selected from down group: PhCH ₂CH ₂C (O)-(N-Me) Val-Tbg-CH ₂-(R)-CH (CH ₂C (O)-CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Et ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃, EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, and EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃.

9, according to the application of the described compositions of claim 5, wherein this nucleoside analog is an acceptable salt on the chemical compound of structural formula 2 or its therapeutics, R in the structural formula 2 ⁹Be hydrogen, hydroxyl or amino.

10, the application of compositions according to claim 5, wherein this nucleoside analog is selected from down group: vidarabine, idoxene, trifluridine, gancyclovir, edoxu-dine, broavir, fiacitabine, penciclovir, famciclovir and roc-iclovir.

11, the application of compositions according to claim 5, wherein this peptide derivant is selected from down group PhCH ₂CH ₂C (O)-(N-Me) Val-Tbg-CH ₂-(R)-CH (CH ₂C (O)-CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Et ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃, EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, and EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃And wherein this nucleoside analog is an acycloguanosine.

12, the application of a kind of peptide derivant in a kind of medicament of preparation, described medicament is used for the treatment of the herpes simplex viral infections of mammiferous tool acycloguanosine resistance, wherein this peptide derivant is an acceptable salt on the chemical compound of structural formula 1 or its therapeutics, and structural formula (1) is A-B-D-CH ₂CH{CH ₂C (O) R ¹C (O)-NHCH{CR ²(R ³) COOH}C (O)-E

1 wherein A be phenylacetyl group, hydrocinnamoyl, (4-aminophenyl) propiono, (4-fluorophenyl) propiono, (4-hydroxy phenyl) propiono, (4-methoxyphenyl)-propiono, 2-(benzyl)-3-hydrocinnamoyl, 2-{ (4-fluorophenyl) methyl }-3-(4-fluorophenyl) propiono, 2-{ (4-methoxyphenyl) methyl }-3-(4-methoxyphenyl) propiono or benzylamino carbonyl; B is (N-Me)-Val or (N-Me)-Ile; Or A and B form a kind of saturated alkyl amino-carbonyl together, it is selected from following groups: the butyl amino carbonyl, 1-Methylethyl amino carbonyl, 1-methyl-propyl amino carbonyl, 1-ethyl propyl amino carbonyl, 1,1-dimethylbutyl amino carbonyl, 1-ethyl-butyl amino carbonyl, 1-propyl group butyl amino carbonyl, 1-ethyl pentyl group amino carbonyl, 1-butyl amyl group amino carbonyl, 1-ethyl-butyl amino carbonyl, 2-ethyl pentyl group amino carbonyl, 1-methyl isophthalic acid-propyl group butyl amino carbonyl, 1-ethyl-1-propyl group butyl amino carbonyl, 1,1-dipropyl butyl amino carbonyl, (1-propyl group cyclopenta) amino carbonyl and (1-propyl group cyclohexyl) amino carbonyl; D is Val, Ile or Tbg; R ¹Be the 1-Methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 1,1-dimethyl propyl, 2,2-dimethyl propyl, cyclobutyl, cyclopenta, cyclohexyl, 1-methylcyclopentyl, NR ⁴R ⁵R wherein ⁴Be hydrogen or low alkyl group, and R ⁵Be low alkyl group, perhaps R ⁴And R ⁵Form pyrrolidinyl with its bonded nitrogen-atoms, piperidino, morpholino or 4-methyl isophthalic acid-piperazinyl; R ²Be hydrogen, R ³Be methyl, ethyl, the 1-Methylethyl, 1, the 1-dimethyl ethyl, propyl group, 2-acrylic or benzyl carry R ²And R ³Carbon atom have (R) configuration, perhaps R ²And R ³Be separately methyl or ethyl, perhaps R separately ²And R ³Form cyclobutyl with its bonded carbon atom, cyclopenta, or cyclohexyl; And E is NHR ⁶R wherein ⁶Be the 2-methyl-propyl, 2,2-dimethyl propyl, 1 (R), 2,2-trimethyl propyl group, 1,1,2,2-tetramethyl propyl group, 1 (R)-ethyl-2,2-dimethyl propyl, 2 (R, S)-methyl butyl, 2, the 2-dimethylbutyl, 3,3-dimethylbutyl, 1 (R), 2,2-trimethyl butyl, 1 (R), 3,3-trimethyl butyl, the 2-ethyl-butyl, 2,2-diethyl butyl, 2-ethyl-1 (R)-methyl butyl, 2-ethyl-2-methyl butyl, 1 (R)-ethyl-3, the 3-dimethylbutyl, 2,2-dimethyl amyl group, cis or trans-2-methylcyclohexyl, 2,2-Dimethylcyclohexyl or cyclohexyl methyl; Perhaps E is NHCH (R ⁷)-Z wherein carries R ⁷Carbon atom have (S) configuration, R ⁷Be 1, the 1-dimethyl ethyl, the 1-methyl-propyl, the 2-methyl-propyl, 2,2-dimethyl propyl or cyclohexyl methyl, and Z is CH ₂OH, C (O) OH, C (O) NH ₂Or C (O) OR ⁸R wherein ⁸Be methyl, ethyl or propyl group.

13, the application of peptide derivant according to claim 12, wherein this peptide derivant is an acceptable salt on the chemical compound of structural formula 1 or its therapeutics, A is a hydrocinnamoyl in the structural formula 1,2-(benzyl)-3-hydrocinnamoyl or benzylamino carbonyl; B is (N-Me) Val; D is Tbg; R ¹Be the 1-Methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 1,1-dimethyl propyl, 2,2-dimethyl propyl, cyclobutyl, cyclopenta, cyclohexyl or 1-methylcyclopentyl; R ²Be hydrogen, R ³Be methyl, ethyl, the 1-Methylethyl, propyl group or benzyl, and carry R ²And R ³Carbon atom have (R) configuration, perhaps R ²And R ³Be separately methyl or ethyl, perhaps R separately ²And R ³Form cyclobutyl with its bonded carbon atom, cyclopenta or cyclohexyl; E is NHR ⁶, R wherein ⁶Be 2, the 2-dimethyl propyl, 1 (R), 2,2-trimethyl propyl group, 1 (R)-ethyl-2, the 2-dimethyl propyl, 2,2-dimethylbutyl or 1 (R)-ethyl-3,3-dimethylbutyl or E are NHCH (R ⁷)-Z wherein carries R ⁷Carbon atom have (S) configuration, R ⁷Be 2,2-dimethyl propyl, Z are CH ₂OH, C (O) OH, C (O) NH ₂Or C (O) OR ⁸R wherein ⁸Be methyl, ethyl or propyl group.

14, the application of peptide derivant according to claim 12, wherein this peptide derivant is an acceptable salt on the chemical compound of structural formula 1 or its materia medica, wherein A and B form a kind of saturated alkyl amino-carbonyl together in the structural formula 1, it is selected from down group: 1-ethyl propyl amino carbonyl, 1-ethyl-butyl amino carbonyl, 1-propyl group butyl amino carbonyl, 2-ethyl pentyl group amino carbonyl, 1-methyl isophthalic acid-propyl group butyl amino carbonyl, 1-ethyl-1-propyl group butyl amino carbonyl, 1,1-dipropyl butyl amino carbonyl and (1-propyl group cyclopenta) amino carbonyl; D is Tbg; R ¹Be the 1-Methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 1,1-dimethyl propyl, 2,2-dimethyl propyl, cyclobutyl, cyclopenta, cyclohexyl or 1-methylcyclopentyl; R ²Be hydrogen and R ³Be methyl, ethyl, the 1-Methylethyl, propyl group or benzyl carry R ²And R ³Carbon atom have (R)-configuration, perhaps R ²And R ³Be separately methyl or ethyl, perhaps R separately ²And R ³Form cyclobutyl with its bonded carbon atom, cyclopenta or cyclohexyl; E is NHR ⁶R wherein ⁶Be 2, the 2-dimethyl propyl, 1 (R), 2,2-trimethyl propyl group, 1 (R)-ethyl-2, the 2-dimethyl propyl, 2, the 2-dimethylbutyl, perhaps 1 (R)-ethyl-3,3-dimethylbutyl or E are NHCH (R ⁷)-Z wherein carries R ⁷Carbon atom have (S) configuration, R ⁷Be 2,2-dimethyl propyl, Z are CH ₂OH, C (O) OH, C (O) NH ₂Perhaps C (O) OR ⁸, R wherein ⁸Be methyl, ethyl or propyl group.

15, according to the application of the described peptide derivant of claim 12, wherein this peptide derivant is selected from down group: PhCH ₂CH ₂C (O)-(N-Me) Val-Tbg-CH ₂-(R)-CH (CH ₂C (O)-CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Et ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃, EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, and EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃.

16, acceptable salt and a kind of antiviral nucleoside analogue on the described peptide derivant of claim 12 or its therapeutics, or the compositions of acceptable salt on its therapeutics, be used to prepare a kind of application of medicament, this medicament is used for the treatment of the herpes simplex viral infections of mammiferous tool acycloguanosine resistance.

17, the application of compositions according to claim 16, wherein this peptide derivant is an acceptable salt on the peptide derivant of structural formula 1 or its therapeutics, A is a hydrocinnamoyl in the structural formula 1,2-(benzyl)-3-hydrocinnamoyl or benzylamino carbonyl; B is (N-Me) Val; D is Tbg; R ¹Be the 1-Methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 1,1-dimethyl propyl, 2,2-dimethyl propyl, cyclobutyl, cyclopenta, cyclohexyl or 1-methylcyclopentyl; R ²Be hydrogen and R ³Be methyl, ethyl, the 1-Methylethyl, propyl group or benzyl carry R ²And R ³Carbon atom have (R)-configuration, perhaps R ²And R ³Independently be methyl or ethyl, perhaps R separately ²And R ³Form cyclobutyl with its bonded carbon atom, cyclopenta or cyclohexyl; And E is NHR ⁶R wherein ⁶Be 2, the 2-dimethyl propyl, 1 (R), 2,2-trimethyl propyl group, 1 (R)-ethyl-2, the 2-dimethyl propyl, 2,2-dimethylbutyl or 1 (R)-ethyl-3,3-dimethylbutyl or E are NHCH (R ⁷)-Z wherein carries R ⁷Carbon atom have (S) configuration, R ⁷Be 2,2-dimethyl propyl, Z are CH ₂OH, C (O) OH, C (O) NH ₂Perhaps C (O) OR ⁸, R wherein ⁸Be methyl, ethyl or propyl group.

18, according to the application of the described compositions of claim 16, wherein this peptide derivant is an acceptable salt on the chemical compound of structural formula 1 or its therapeutics, A and B form a kind of saturated alkyl amino-carbonyl together in the structural formula 1, it is selected from down group: 1-ethyl propyl amino carbonyl, 1-ethyl-butyl amino carbonyl, 1-propyl group butyl amino carbonyl, 2-ethyl pentyl group amino carbonyl, 1-methyl isophthalic acid-propyl group butyl amino carbonyl, 1-ethyl-1-propyl group amino carbonyl, 1,1-dipropyl butyl amino carbonyl and (1-propyl group cyclopenta) amino carbonyl; D is Tbg; R ¹Be the 1-Methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 1,1-dimethyl propyl, 2,2-dimethyl propyl, cyclobutyl, cyclopenta, cyclohexyl or 1-methylcyclopentyl; R ²Be hydrogen and R ³Be methyl, ethyl, the 1-Methylethyl, propyl group or benzyl, and carry R ²And R ³Carbon atom have (R) configuration, perhaps R ²And R ³Independently be methyl or ethyl, perhaps R separately ²And R ³Form cyclobutyl with its bonded carbon atom, cyclopenta or cyclohexyl; E is NHR ⁶, R wherein ⁶Be 2, the 2-dimethyl propyl, 1 (R), 2,2-trimethyl propyl group, 1 (R)-ethyl-2, the 2-dimethyl propyl, 2,2-dimethylbutyl or 1 (R)-ethyl-3,3-dimethylbutyl or E are NHCH (R ⁷)-Z wherein carries R ⁷Carbon atom have (S) configuration, R ⁷Be 2,2-dimethyl propyl, Z are CH ₂OH, C (O) OH, C (O) NH ₂Perhaps C (O) OR ⁸, R wherein ⁸Be methyl, ethyl or propyl group.

19, the application of compositions according to claim 16, wherein this peptide derivant is selected from down group: PhCH ₂CH ₂C (O)-(N-Me) Val-Tbg-CH ₂-(R)-CH (CH ₂C (O)-CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Et ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃, EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, and EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃.

20, according to the application of the described compositions of claim 16, wherein this nucleoside analog is an acceptable salt on the chemical compound of structural formula 2 or its therapeutics, R in the structural formula 2 ⁹Be hydrogen, hydroxyl or amino.

21, according to the application of the described compositions of claim 16, wherein this nucleoside analog is selected from down group: vidarabine, idoxene, trifluridine, gancyclovir, edoxudi-ne, broavir, fiacitabine, penciclovir, famciclovir and rocic-lovir.

22, according to the application of the described compositions of claim 16, wherein this peptide derivant is selected from down group: PhCH ₂CH ₂C (O)-(N-Me) Val-Tbg-CH ₂-(R)-CH (CH ₂C (O)-CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Et ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃, EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, and EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃Wherein this nucleoside analog is an acycloguanosine.

23, a kind of pharmaceutical composition for the treatment of the herpes simplex viral infections of tool acycloguanosine resistance comprises the peptide derivant of the structural formula (1) as active component, or can accept carrier on acceptable salt and a kind of materia medica on its therapeutics

A-B-D-CH ₂CH{CH ₂C(O)R ¹}C(O)-NHCH{CR ²(R ³)COOH}C(O)-E

1 wherein A be phenylacetyl group, hydrocinnamoyl, (4-aminophenyl) propiono, (4-fluorophenyl) propiono, (4-hydroxy phenyl) propiono, (4-methoxyphenyl)-propiono, 2-(benzyl)-3-hydrocinnamoyl, 2-{ (4-fluorophenyl) methyl }-3-(4-fluorophenyl) propiono, 2-{ (4-methoxyphenyl) methyl }-3-(4-methoxyphenyl) propiono or benzylamino carbonyl; B is (N-Me)-Val or (N-Me)-Ile; Or A and B form a kind of saturated alkyl amino-carbonyl together, it is selected from following groups: the butyl amino carbonyl, 1-Methylethyl amino carbonyl, 1-methyl-propyl amino carbonyl, 1-ethyl propyl amino carbonyl, 1,1-dimethyl ethyl butyl amino carbonyl, 1-ethyl-butyl amino carbonyl, 1-propyl group butyl amino carbonyl, 1-ethyl pentyl group amino carbonyl, 1-butyl amyl group amino carbonyl, 1-ethyl-butyl amino carbonyl, 2-ethyl pentyl group amino carbonyl, 1-methyl isophthalic acid-propyl group butyl amino carbonyl, 1-ethyl-1-propyl group butyl amino carbonyl, 1,1-dipropyl butyl amino carbonyl, (1-propyl group cyclopenta) amino carbonyl, and (1-propyl group cyclohexyl) amino carbonyl; D is Val, Ile or Tbg; R ¹Be the 1-Methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 1,1-dimethyl propyl, 2,2-dimethyl propyl, cyclobutyl, cyclopenta, cyclohexyl, 1-methylcyclopentyl, NR ⁴R ⁵R wherein ⁴Be hydrogen or low alkyl group, and R ⁵Be low alkyl group, perhaps R ⁴And R ⁵Form pyrrolidinyl with its bonded nitrogen-atoms, piperidino, morpholino or 4-methyl isophthalic acid-piperazinyl; R ²Be hydrogen, R ³Be methyl, ethyl, the 1-Methylethyl, 1, the 1-dimethyl ethyl, propyl group, 2-acrylic or benzyl carry R ²And R ³Carbon atom have (R)-configuration, perhaps R ²And R ³Be methyl or ethyl, perhaps R independently of one another ²And R ³Form cyclobutyl with its bonded carbon atom, cyclopenta, or cyclohexyl; And E is NHR ⁶R wherein ⁶Be the 2-methyl-propyl, 2,2-dimethyl propyl, 1 (R), 2,2-trimethyl propyl group, 1,1,2,2-tetramethyl propyl group, 1 (R)-ethyl-2,2-dimethyl propyl, 2-(R, S)-methyl butyl, 2, the 2-dimethylbutyl, 3,3-dimethylbutyl, 1 (R), 2,2-trimethyl butyl, 1 (R), 3,3-trimethyl butyl, the 2-ethyl-butyl, 2,2-diethyl butyl, 2-ethyl-1 (R)-methyl butyl, 2-ethyl-2-methyl butyl, 1 (R)-ethyl-3, the 3-dimethylbutyl, 2,2-dimethyl amyl group, cis or trans-2-methylcyclohexyl, 2,2-Dimethylcyclohexyl or cyclohexyl methyl; Perhaps E is NHCH (R ⁷The carbon atom that)-Z wherein carries R7 has (S) configuration, R ⁷Be 1, the 1-dimethyl ethyl, the 1-methyl-propyl, the 2-methyl-propyl, 2,2-dimethyl propyl or cyclohexyl methyl, and Z is CH ₂OH, C (O) OH), C (O) NH ₂Perhaps C (O) OR ⁸R wherein ⁸Be methyl, ethyl or propyl group.

24, pharmaceutical composition according to claim 23, wherein this active component comprises acceptable salt on the peptide derivant of structural formula 1 of claim 23 definition or its therapeutics, with the compositions that acceptable salt on a kind of antiviral nucleoside analogue or its therapeutics is formed, be used for the treatment of the herpes simplex viral infections of tool acycloguanosine resistance.

25, pharmaceutical composition according to claim 24, wherein this peptide derivant is selected from down group: PhCH ₂CH ₂C (O)-(N-Me) Val-Tbg-CH ₂-(R)-CH (CH ₂C (O)-CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Et ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-γ MeLeucinol, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, Pr ₂CHNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃, EtPr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NH-(R)-CH (Et) CMe ₃, and Etpr ₂CNHC (O)-Tbg-CH ₂-(R)-CH (CH ₂C (O) CMe ₃) C (O)-Asp (cyPn)-NHCH ₂CMe ₃Wherein this nucleoside analog is an acycloguanosine.

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