CN1315963A - Benzodiazepines and benzothiazepines derivatives and HBSAG peptides binding to annexins, their compositions and use - Google Patents

Benzodiazepines and benzothiazepines derivatives and HBSAG peptides binding to annexins, their compositions and use Download PDF

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CN1315963A
CN1315963A CN99810388A CN99810388A CN1315963A CN 1315963 A CN1315963 A CN 1315963A CN 99810388 A CN99810388 A CN 99810388A CN 99810388 A CN99810388 A CN 99810388A CN 1315963 A CN1315963 A CN 1315963A
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seq
annexin
bio
protein
peptide
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E·德普拉
H·梅雷尔斯
G·梅尔滕斯
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Fujirebio Europe NV SA
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Innogenetics NV SA
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • A61P31/22—Antivirals for DNA viruses for herpes viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10—Tetrapeptides
    • C07K5/1002—Tetrapeptides with the first amino acid being neutral
    • C07K5/1005—Tetrapeptides with the first amino acid being neutral and aliphatic
    • C07K5/1008—Tetrapeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atoms, i.e. Gly, Ala
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10—Tetrapeptides
    • C07K5/1019—Tetrapeptides with the first amino acid being basic
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/16011—Herpesviridae
    • C12N2710/16711—Varicellovirus, e.g. human herpesvirus 3, Varicella Zoster, pseudorabies
    • C12N2710/16722—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2730/00—Reverse transcribing DNA viruses
    • C12N2730/00011—Details
    • C12N2730/10011—Hepadnaviridae
    • C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
    • C12N2730/10122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/16011—Orthomyxoviridae
    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
    • C12N2760/16122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
    • G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
    • G01N2333/4701—Details
    • G01N2333/4718—Lipocortins

Abstract

The present invention relates to 1,4-benzodiazepines or 1,4-benzothiazepines derivatized with a peptide that can inhibit the interaction between annexin and annexin binding proteins. In particular, the present invention relates to 1,4-benzodiazepines or 1,4-benzothiazepines derivatives that can inhibit the interaction between annexin and viral proteins that bind annexins such as the HBaAg protein of HBV, glycoprotein B of the cytomegalovirus or any annexin binding protein from the influenza virus. These 1,4-benzodiazepines or 1,4-benzothiazepines derivates can be used to prevent or treat diseases in which interactions between annexin family members and annexin binding proteins are involved such as HBV and/or HBV infections, cytomegalovirus infections or influenza virus infections.

Description

Binding film joins proteic benzodiazepine class and Benzothiazepine derivatives and HBSAG peptide, its composition and purposes
Invention field
The present invention relates to finds to derive 1 of peptide chain, and 4-benzodiazepine class and 1,4-Benzothiazepine class suppress the combination of HbsAg to annexin V.Therefore the present invention relates to such molecule and treatment hepatitis B virus and hepatitis delta viral infection thereof or wherein relate to protein and the purposes of interactional any other disease of annexin family member.
Background of invention
Hepatitis B virus (HBV) belongs to hepadnavirus, it is characterized in that become significantly liver property and species specificity.Hepatitis D virus (HDV) is represented the naturally occurring subvirus satellite (Rizetto etc., 1986) of HBV.HBV causes main medical problem, for example chronic hepatopathy and liver cell cancer (Schroder and Zentgraf, 1990).Infect with independent HBV that to compare the HDV superingection more serious usually.Estimate that the whole world has 300 million peoples to carry this virus, in the U.S. 70,000 carrier's co-infected HDV are just arranged simultaneously.
In the HBV genome, there is the native sequences variation in more particularly in the S gene (also referring to following paragragh).Be basic design A to the F genotype of HBV (about summarizing) with this sequence difference percentage referring to Magnius and Norder, 1995.
Tunicle HBV is made up of three kinds of relevant glycoprotein, they are called hepatitis B surface antigen(HBsAg) (HBsAg), it is the product of S gene: 1) " little " transmembrane protein, also be referred to as major protein or little S-albumen, form by 226 amino acid (aa), 2) " in " protein, it comprises little S-albumen and at 55 other amino acid corresponding to the preceding-S2 district of S gene of N-end, with 3) " greatly " protein is by forming corresponding to 389 or 400 amino acid distinguishing below: before the S+-S2+ before-S1 (the terminal aa of 108-119N-) (Heerman etc., 1984; Robinson etc., 1987).The tunicle of HDV is also derived by HBV fully and mainly by little HBsAg, HBsAg forms among the 5-10%, does not have big HBsAg or big HBsAg to be less than 1% (Bonino etc., 1986).
Influenza virus causes the recurrent prevailing disease, or even global infectious disease.Absence from duty during these outbreaks of communicable diseases causes the tremendous economic loss and follows admission rate to increase, even causes coming from the death (about summarizing referring to Cox and Fukuda, 1998) of respiratory disease.
Cytomegalovirus (cytomegalovirus) is a member of simplexvirus family and is for example AIDS patient's important pathogenic agent of non-responsiveness individuality.In the coating of simplexvirus, there is the protein that is referred to as Glycoprotein B.Prove that this protein is essential for infection, described infection comprises and enters the propagation (Cai etc., 1988) to cell of host cell and cell.
Annexin V (also being referred to as the endonexin II, placenta anticoagulant protein, PP4 or lipocortin V) is Ca relevant on the structure 2+A member of-dependency phosphatide-conjugated protein family is known as annexin (Klee, 1988 of molecular weight between 32-67kDa; Zaks and Creutz, 1990).In various tissues, find the annexin V, liver for example, spleen, lung, intestines and placenta (Walker etc., 1990).This protein was once described with Ca 2+-dependency mode is in conjunction with placental membrane (Haigler etc., 1987) and vitro inhibition blood coagulation (Grundmann etc., 1988) and Phospholipase A2 activity (Pepinsky etc., 1988).Other researchist proves that the behavior of annexin V resembles conformity membrane albumen and forms calcium-selectivity cationic channel (Rojas etc., 1990; Bianchi etc., 1992).
Prove 1 recently, 4-benzodiazepine class (Hofmann etc., 1998) and 1, (Kaneko etc., 1997a b) can be in conjunction with the annexin V for 4-Benzothiazepine class.The annexin II is relevant with the annexin V and also in conjunction with 1,4-benzodiazepine class (Hofmann etc., 1998).Annexin also plays a large amount of and the effect of non-virus protein bonded, and described non-virus protein is collagen protein for example, cytotactin, Actin muscle, synapsin, calspectin, insulin receptor, Profibrinolysin or the like (about summarizing) referring to Sheldon and Chen, 1996.
The annexin V that exists on our the nearest reference liver plasma membrane is with Ca 2+-dependency mode specificity is in conjunction with " little " HBsAg (Hertogs etc., 1993; WO94/01554).With the rabbit of natural human liver annexin V or the immunity of reorganization annexin V, perhaps with the F of the anti-annexin V IgG of rabbit (ab ') 2Spontaneous anti--the Te that produces specific recognition HBsAg of the chicken of-fragment immunity answers the discovery of antibody (Ab2) further to support the receptor-ligand between HBsAg and the annexin V to concern (Hertogs etc., 1994).We prove that also the HBsAg of HDV particle by comprising the HBV coating is in conjunction with annexin V (de Bruin etc., 1994).Position, site on the HBsAg that relates in the combination of annexin V has been described among the WO97/07268.These sites are made up of 100-160 the amino acid of " little " HBsAg in this zone, estimate that it is positioned at this viral outside surface (relevant summary referring to Berting etc., 1995).
Combine (Pietropaolo and Compton, 1997) of the combining of influenza virus and annexin V (Huang etc., 1996) and cytomegalovirus and annexin II have also been proved.
From above-mentioned document clear learn that HBV and/or HDV infect and other virus infection for example influenza and cytomegalovirus infection still are major health.Therefore developing and disturbing the new medicine of HBV and/or HDV or these other viral life cycle still is a main demand.Being suppressed a kind of like this mode binding film with HBV and/or HDV or these other virus and combining of annexin, to join proteic compound may be the good candidate that disturbs these viral life cycles, therefore may be as the good candidate for the treatment of medicine for treating viral infections.Therefore, pressing for provides and the annexin specific effect, suppresses HBV and/or HDV or these other virus and the interactional molecule of annexin simultaneously.
Purpose of the present invention
The objective of the invention is to suppress the protein-bonded interaction of annexin and annexin.
The present invention's purpose more specifically is the interaction that inhibition annexin and energy binding film join proteic viral protein.
The present invention another more specifically purpose be to suppress the interaction of the surface antigen of annexin V and HBV.
The present invention another more specifically purpose be to suppress the interaction of the Glycoprotein B of annexin II and cytomegalovirus.
The present invention another more specifically purpose be to suppress annexin V and protein interactions derived from any strains of influenza viruses that combines the annexin V.
Another object of the present invention is that prevention or treatment wherein relate to annexin family member and the annexin interactional any disease between conjugated protein.
The present invention's purpose more specifically is that prevention or treatment wherein relate to the interactional any virus disease between annexin family member and the viral protein.
The present invention another more specifically purpose be that prevention or treatment HBV and/or HDV infect.
The present invention another more specifically purpose be the prevention or the treatment cytomegalovirus infection.
The present invention another more specifically purpose be the prevention or the treatment influenza infection.
Think and realize all these purposes of the present invention by following embodiment.
Detailed description of the present invention
The present invention described herein quotes the previous disclosed work and the patent application of not winding up the case.For instance, such work is by scientific and technical literature, and the patent or the patent application of not winding up the case are formed.Above or all these publications of hereinafter quoting and the application incorporated by reference here.
The present invention relates to derive has 1 of one or more peptides, 4-benzodiazepine class and 1,4-Benzothiazepine class.
Specifically, the present invention relates to derive and have one or more peptides to make this derivative suppress interactional 1 between conjugated protein of annexin and some annexins, 4-benzodiazepine class and 1,4-Benzothiazepine class.
Molecule--1 of the annexin V that relates in entering in conjunction with HBV virus, 4-benzodiazepine class and 1,4-Benzothiazepine class does not suppress the combination of HBV to the annexin V.Interaction between the HBsAg of the application annexin V and HBV during based on these compound deriving peptide parts can repressed surprising discovery.These are new 1 years old, therefore 4-benzodiazepine class and 1, the derivative of 4-Benzothiazepine class are to disturb HBV and/or HDV and in conjunction with annexin in general or new leading in the drug discovery of other viral life cycle of annexin V more particularly.These molecules generally can disturb annexin and another kind of proteinic any interaction.By with 1,4-benzodiazepine class or 1,4-Benzothiazepine class selective derivatizationization, can with particular approach lead these interactional any.
Term " annexin " (also being referred to as endonexin II, placenta anticoagulant protein, PP4 or lipocortin V) refers to Ca relevant on the structure 2+Any member of-dependency phosphatide-conjugated protein family is known as annexin (Klee, 1988 of molecular weight between 32-67kDa; Zaks and Creutz, 1990).More particularly, described annexin can be to show in conjunction with the HBsAg of HBV with in conjunction with the annexin V of influenza virus, the annexin II of perhaps known Glycoprotein B in conjunction with cytomegalovirus.But specified annexin is not limited to these annexins among the present invention, and can be and all other annexins of the conjugated protein bonded of annexin.
Described 1,4-benzodiazepine or 1,4-Benzothiazepine can have any modification well known in the art, for example those that mention among W094/11360 and the W092/12148.Preferably, 1,4-benzodiazepine or 1,4-Benzothiazepine have structure shown in the formula I (Fig. 1), wherein:
* at 1 :-Benzothiazepine class has a sulphur atom (X represents S); X also can be SO n, n=0 wherein, 1,2;
-benzodiazepine class has a nitrogen-atoms (X represents N), and it is modified with carboxymethyl (CH in addition at 1 2COOH) afterwards, allow by the C-of peptide terminal that connect or by the N-of peptide terminal connect be modified with in claims peptide or its part that is defined as " containing the peptide of a protein-bonded annexin of annexin " in addition in conjunction with epi-position.Be not modified with at N under the situation of a peptide, modification can be H, alkyl, and phenyl ,-COZ, wherein Z represents H, alkyl, the phenyl of phenyl or replacement;
* 1,2,3,4,5 can form two keys with adjoining position; If this is the case, side chain R then 2, R 4, R 5Or R 7Do not exist;
* the R on 2 and/or 2 ' 1And/or R 2Can represent H, amine, alkyl or 2 can be oxidized (=O).At R 1And/or R 2Be under the situation of amine, it can be further or directly connects or to be connected in the N-of peptide terminal and be defined as peptide or its part of " containing the peptide of a protein-bonded annexin of annexin in conjunction with epi-position " in claims and replace by resembling the such linking group of glutaraldehyde or succinyl oxide by the C-of peptide is terminal;
* the R on 3 and/or 3 ' 3And/or R 4Can represent H, amine or alkyl.At R 3And/or R 4Be under the situation of amine, it can be further or directly connects or to be connected in the N-of peptide terminal and be defined as peptide or its part of " containing the peptide of a protein-bonded annexin of annexin in conjunction with epi-position " in claims and replace by resembling the such linking group of glutaraldehyde or succinyl oxide by the C-of peptide is terminal;
* the R on 4 5Can represent 4 to be modified with carboxymethyl (CH in addition 2COOH) back or directly terminal that connect or by being defined as the peptide of " containing the peptide of a protein-bonded annexin of annexin " or the side chain of its part in the terminal claims that connect of the N-of peptide in conjunction with epi-position by the C-of peptide.At R 5Not under the situation of the peptide that defines in claims, R 5Can represent H, alkyl ,-CO-R 12Perhaps 3-(1-(4-benzyl) piperidyl) propionyl);
* R 6And/or R 7Represent H or alkyl or phenyl, may further be substituted with alkyl, cyano group, halogen, nitro, alkyl alkoxy, alkyloyl, carboxyl, alkyloyl alkoxyl group, formamyl;
* R 8, R 9, R 10And/or R 11Represent H, alkyl, cyano group, halogen, nitro, alkyl alkoxy, alkyloyl, carboxyl, alkyloyl alkoxyl group, formamyl;
* R 12Represent H or alkyl or may further be substituted with alkyl, cyano group, halogen, nitro, alkyl alkoxy, alkyloyl, carboxyl, alkyloyl alkoxyl group, the phenyl of formamyl.
Be used to prepare 1 of molecule of the present invention, 4-benzodiazepine or 1,4-Benzothiazepine can also be any pharmaceutically acceptable salts of this structural formula.Be used to prepare 1 of molecule of the present invention, 4-benzodiazepine can be all derivatives well known in the art, such as but not limited to alprazolam, and Bromazepam, chloordiazepoxide, clobazam, clonazepam, chlordiazepoxide, diazepam, fludiazepam, flunitrazepam, flurazepam, lorazepam, nitrazepam, oxazepam, temazepam, triazolam, BDA250, BDA452, BDA753 (also referring to Fig. 6 and Hoffman etc., 1998).But compound of the present invention is not BDA753, a kind of compound (FR2479818 according to the formula II; Fig. 2) or a kind of compound (Nachman etc., 1998 according to the formula III; Fig. 3).1, the 4-Benzothiazepine also can be any known derivative in this area, such as but not limited to K-201 (Kaneko etc., 1997a).
According to the present invention, these 1,4-benzodiazepine or 1, the 4-Benzothiazepine is 2, and deriving after 2 ', 3 and/or 3 ' (in the formula I) is modified with amine has a peptide species, they can be 2, and 2 ', 3 and/or 3 ' (in the formula I) connects (two peptides can connect in identical position).This peptide can or directly pass through the terminal amine that connects of C-of this peptide, and perhaps this peptide can pass through terminal connection of N-of this peptide by using a linking group.
Described linking group can be any linking group that two amine residues are connected.Preferred described linking group is a dialdehyde, for example glutaraldehyde or acid anhydrides, for example succinyl oxide.Other possible linking group also can (Rockford, IL USA) provide by Pierce.Fig. 9 and Figure 10 have provided by utilizing a linking group to make peptide and 1, the embodiment of 4-benzodiazepine connection basis reaction.
Described polypeptide also can be connected to 1 and/or 4 (in the formula I).In this case, described peptide or the C-by peptide are terminal connects or is modified with carboxymethyl (CH respectively in addition at 1 and/or 4 2COOH) this peptide connects by N-is terminal afterwards.
In a preferred embodiment, peptide with at 5 places by phenyl replace 1,1 and/or 3 connection of 4-benzodiazepine .A peptide, R, can be by its C-terminal and described 3 be modified with 1 after the amine, 3 connections of 4-benzodiazepine , perhaps, a peptide, R ', can be by its N-terminal and described 1 be modified with 1 after the carboxymethyl, 1 connection of 4-benzodiazepine , perhaps, two peptides, R and R ', respectively with described 1,3 of 4-benzodiazepine is connected with 1.Therefore, the present invention relates to described deriving has one or two peptide, R and/or R ', 1,4-benzodiazepine , R, R ' and/or R+R ' representative comprises peptide or its part of the protein-bonded annexin of annexin in conjunction with epi-position.Described 1,4-benzodiazepine has the structure shown in the formula IV (Fig. 4) then, wherein:
* R, R ' and/or R+R ' constitute and comprise peptide or its part of the protein-bonded annexin of annexin in conjunction with epi-position.Having only R is under the situation of a peptide, and 1 is methylated or carboxymethylated.Having only R ' is under the situation of a peptide, and 3 contain amine;
* X representative can be by-COCH 3The N-end of the peptide of modifying;
* Y representative can be by-NH 2Or-the C-end of the peptide of Methionin-biotin modification.
Be used for by Fmoc describedly 1 for the peptide synthetic on basis, the synthetic structural unit of 4-benzodiazepine derivative can obtain from Neosystem (Strasbourg, France).
In another embodiment of the invention, by using aforesaid linking group, peptide, R, terminal and described 1 by its N-, 3 connections of 4-benzodiazepine (being replaced by phenyl) at 5.In this case, 1 nitrogen-atoms (formula I) is methylated or carboxymethylation.Therefore the present invention relates to derive have and comprise the peptide of the protein-bonded annexin of annexin, R, the perhaps compound according to the formula V of its part in conjunction with epi-position.
With 1,4-benzodiazepine or 1, the polypeptide that the 4-Benzothiazepine connects can be that all comprising from known binding film joins the polypeptide of the annexin of proteic protein derived in conjunction with epi-position.
Term " annexin is conjugated protein " refers to form with annexin all proteins of mixture.
Term " annexin is in conjunction with epi-position " refer to the annexin specificity interact to form annexin conjugated protein-the protein-bonded district of annexin of annexin mixture." annexin is in conjunction with epi-position " comprises at least 2, preferred 3,4,5,6,7,8,9,10,11,12,13,14,15,20,25,30,35 or more a plurality of amino acid whose annexin conjugated protein.Can determine epi-position or pass through various computer attribute models indication well known in the art by any technology well known in the art.
Term " polypeptide " or " peptide " can exchange use, refer to and its identical or close polymer of amino acid of the proteinic part of deutero-therefrom.Described polypeptide can be made up of multiple amino acids, and preferably, polypeptide is by being less than 13 amino acid, and is preferred 12,11,10,9,8,7,6,5, and 4,3 or 2 amino acid are formed, and such annexin is included in this polypeptide in conjunction with epi-position.Under The compounds of this invention has situation according to the structure of formula IV, peptide R or R ' only are made up of 1 amino acid and/or annexin to be included in conjunction with epi-position also be possible in the polypeptide of representing R+R '.Should notice that from the polypeptide of the land of the conjugated protein deutero-of annexin and formation and annexin may be the part of conformation land, therefore can not form by the protein-bonded approaching aminoacid sequence of annexin.
By any method well known in the art, for example, the classical chemosynthesis as Houbenweyl (1974) and Atherton and Shepard (1989) description perhaps utilizes the recombinant DNA technology of describing as (1982) such as Maniatis, can prepare polypeptide.Term " polypeptide " does not refer to not get rid of the posttranslational modification of polypeptide yet, for example glycosylation, and acylations, phosphorylation is with fatty acid modifying or the like.Comprise in this definition and for example contain one or more amino acid analogues polypeptide of (comprising alpha-non-natural amino acid or D-isomeric form), polypeptide with substituted key, the sudden change version of this polypeptide or native sequences variant are (for example, as mentioned above corresponding to the proteinic peptide variant of HBsAg of the genotypic HBV of A to F of HBV), the polypeptide that disulfide linkage is arranged in the middle of the cysteine residues, oppositely-(for example Guichard etc. is described for counter-rotating (reverso-inverso) polypeptide, 1994), and other modifications well known in the art.
In a specific embodiments, join proteic viral protein from known binding film and derive and 1,4-benzodiazepine or 1, the polypeptide that the 4-Benzothiazepine connects.Term " from the viral protein polypeptides derived " refers to polymer of amino acid, and identical or the close and annexin that comprises this viral protein of its part with deutero-viral protein therefrom is in conjunction with epi-position.
In another specific embodiment, from known antigens (Heerman etc., 1984; Robinson etc., 1987) the HBsAg protein derived and 1 of HBV, 4-benzodiazepine or 1, the polypeptide that the 4-Benzothiazepine connects.Here employed term " from the polypeptide of the HBsAg protein derived of HBV " refers to a kind of polypeptide with aminoacid sequence, and identical or the close and annexin that comprises HBsAg of its part with the aminoacid sequence of " little " HBsAg is in conjunction with epi-position.Should be clear " from the polypeptide of the HBsAg protein derived of HBV " (genotype A to F, Magnius and Norder1995) can derive from any genotype of HBV.
In another specific embodiment of the present invention, derive and 1 4-benzodiazepine or 1, the polypeptide that the 4-Benzothiazepine connects from the Glycoprotein B of cytomegalovirus.Term " from the Glycoprotein B polypeptides derived " refers to a kind of polymer of amino acid, and identical or the close and annexin that comprises Glycoprotein B of its part with the Glycoprotein B of cytomegalovirus is in conjunction with epi-position.
In another specific embodiment of the present invention, from protein derived and 1 from any strains of influenza viruses, 4-benzodiazepine or 1, the polypeptide that the 4-Benzothiazepine connects, condition is that this influenza proteins matter is in conjunction with the annexin V.Term " polypeptide of the protein derived of the susceptible strain that always flows automatically " refers to a kind of polymer of amino acid, its with and the annexin that comprise influenza proteins identical or close from the proteinic part of strains of influenza viruses in conjunction with epi-position.
In addition, the object of the invention is to provide has 1 of derived chemotactic peptide effect, 4-benzodiazepine or 1,4-Benzothiazepine, described derived chemotactic peptide comprise as defined above or any combination of the peptide that produces from all mutant strains of HBV and/or HDV, cytomegalovirus or influenza virus.
In a specific embodiment, the invention provides derives 1 of a peptide, 4-benzodiazepine or 1,4-Benzothiazepine molecule, wherein said peptide comprise be less than 13 amino acid of the annexin in one of zone of comprising following HBV surface antigen in conjunction with epi-position:
KTCTTPAQGN(SEQ?ID?NO1)
FAKYLWEWASVR (SEQ ID NO2) once described this and related to (W097/07268 and the present invention) in the interaction of HBsAg and annexin V.
More particularly, of the present invention with 1,4-benzodiazepine or 1, the polypeptide that the 4-Benzothiazepine derivatives connects relate to 122-131 amino acids sequence deutero-2-10 the amino acid whose polypeptide that comprises from the HBsAg of HBV, shown in SEQ ID NO1.More particularly, relate to the peptide that comprises or form by following amino acid (aa) sequence:
Aa122-123, aa123-124, aa124-125, aa125-126, aa126-127, aa127-128, aa128-129, aa129-130, aa130-131, aa122-124, aa123-125, aa124-126, aa125-127, aa126-128, aa127-129, aa128-130, aa129-131, aa122-125, aa123-126, aa124-127, aa125-128, aa126-129, aa127-130, aa128-131, aa122-126, aa123-127, aa124-128, aa125-129, aa126-130, aa127-131, aa122-127, aa123-128, aa124-129, aa125-130, aa126-131, aa122-128, aa123-129, aa124-130, aa125-131, aa122-129, aa123-130, aa124-131, aa122-130, aa123-131, aa122-131. similarly, of the present invention with 1,4-benzodiazepine or 1, the polypeptide that the 4-Benzothiazepine derivatives connects is particularly related to 158-169 amino acids sequence deutero-2-12 the amino acid whose polypeptide that comprises from the HBsAg of HBV, as above shown in the SEQ ID NO2.More particularly, relate to the peptide that comprises or form by following amino acid (aa) sequence:
Aa158-159, aa159-160, aa160-161, aa161-162, aa162-163, aa163-164, aa164-165, aa165-166, aa166-167, aa167-168, aa168-169, aa158-160, aa159-161, aa160-162, aa161-163, aa162-164, aa163-165, aa164-166, aa165-167, aa166-168, aa167-169, aa158-161, aa159-162, aa160-163, aa161-164, aa162-165, aa163-166, aa164-167, aa165-168, aa166-169, aa158-162, aa159-163, aa160-164, aa161-165, aa162-166, aa163-167, aa164-168, aa165-169, aa158-163, aa159-164, aa160-165, aa161-166, aa162-167, aa163-168, aa164-169, aa158-164, aa159-165, aa160-166, aa161-167, aa162-168, aa163-169, aa158-165, aa159-166, aa160-167, aa161-168, aa162-169, aa158-166, aa159-167, aa160-168, aa161-169, aa158-167, aa159-168, aa160-169, aa158-168, aa159-169, aa158-169. more particularly, the present invention relates to derive has 1 of one of following peptide or its part, 4-benzodiazepine :
(FAKYLWEWASVR) 2-KKGK(bio)GA(SEQ?ID?NO4)
FAKYLW-K(bio)(SEQ?ID?NO5)
FAKYLWEW-K(bio)(SEQ?ID?NO6)
FAKYLWEWAS-K(bio)(SEQ?ID?NO7)
FAKYLWEWASVR-K(bio)(SEQ?ID?NO8)
FAKYLW(SEQ?ID?NO9)
FAKYLWEW(SEQ?ID?NO10)
FAKYLWEWAS(SEQ?ID?NO11)
FAKYLWEWASVR(SEQ?ID?NO2)
IGP1362 or its part and BDA250 or BDA452 or its any derivative are connected the specific compound that obtains having the raising in the interaction of blocking-up annexin V and HBsAg.Should notice that not all sequence obtains identical result.For example, just do not suppress combining of annexin V and HBsAg from the sequence of HBsAg (aa115-125) deutero-IGP1363 peptide representative yet.
In another specific embodiment of the present invention, 1,4-benzodiazepine or 1,4-Benzothiazepine derive by the polypeptide of being made up of following aminoacid sequence or its part:
FARFLWEWASVR-K(bio)(SEQ?ID?NO12)
FGKFLWEWASAR-K(bio)(SEQ?ID?NO13)
LGKYLWEWASAR-K(bio)(SEQ?ID?NO14)
FAKFLWEWASVR-K(bio)(SEQ?ID?NO15)
KYGW-K(bio)(SEQ?ID?NO16)
KFGW-K(bio)(SEQ?ID?NO17)
RFGW-K(bio)(SEQ?ID?NO18)
AYLW-K(bio)(SEQ?ID?NO19)
AFLW-K(bio)(SEQ?ID?NO20)
KYLW-K(bio)(SEQ?ID?NO21)
RFLW-K(bio)(SEQ?ID?NO22)
KFLW-K(bio)(SEQ?ID?NO23)
FARFLWEWASVR(SEQ?ID?NO24)
FGKFLWEWASAR(SEQ?ID?NO25)
LGKYLWEWASAR(SEQ?ID?NO26)
FAKFLWEWASVR(SEQ?ID?NO27)
KYGW(SEQ?ID?NO28)
KFGW(SEQ?ID?NO29)
RFGW(SEQ?ID?NO30)
AYLW(SEQ?ID?NO31)
AFLW(SEQ?ID?NO32)
KYLW(SEQ?ID?NO33)
RFLW(SEQ?ID?NO34)
KFLW(SEQ?ID?NO35)
Above-mentioned peptide or its part can be for example as mentioned above 1,2, and 2 ', 3,3 ' and/or 4 and 1,4-benzodiazepine or 1, the 4-Benzothiazepine connects.
In a preferred embodiment, according to 1 of formula IV, 1 (R ') of 4-benzodiazepine and/or 3 (R) position connect described peptides.Adding and (R+R ') the aforesaid peptide of representative or its part of right latter two peptide.Therefore, 1,4-benzodiazepine can be in any position of above-mentioned peptide.For example, when R+R '=FAKYLWEWASVR (SEQ ID NO2), R and/or R ' can connect 1 in every way, 4-benzodiazepine (Bdz):
Bdz-FAKYLWEWASVR:R '=FAKYLWEWASVR (SEQ ID NO2); Or
F-Bdz-AKYLWEWASVR:R=F, R '=AKYLWEWASVR (SEQ ID NO36); Or
FA-Bdz-KYLWEWASVR:R=FA (SEQ ID NO37), R '=KYLWEWASVR (SEQ ID NO38); Or
FAK-Bdz-YLWEWASVR:R=FAK (SEQ ID NO39), R '=YLWEWASVR (SEQ ID NO40); Or
FAKY-Bdz-LWEWASVR:R=FAKY (SEQ ID NO41), R '=LWEWASVR (SEQ ID NO42); Or
FAKYL-Bdz-WEWASVR:R=FAKYL (SEQ ID NO43); R '=WEWASVR (SEQ ID NO44); Or
FAKYLW-Bdz-EWASVR:R=FAKYLW (SEQ ID NO9), R '=EWASVR (SEQ ID NO45); Or
FAKYLWE-Bdz-WASVR:R=FAKYLWE (SEQ ID NO46), R '=WASVR (SEQ ID NO47); Or
FAKYLWEW-Bdz-ASVR:R=FAKYLWEW (SEQ ID NO10), R '=ASVR (SEQ ID NO48); Or
FAKYLWEWA-Bdz-SVR:R=FAKYLWEWA (SEQ ID NO49), R '=SVR (SEQ ID NO50); Or
FAKYLWEWAS-Bdz-VR:R=FAKYLWEWAS (SEQ ID NO11), R '=VR (SEQ ID NO51); Or
FAKYLWEWASV-Bdz-R:R=FAKYLWEWASV (SEQ ID NO52), R '=R; Or
FAKYLWEWASVR-Bdz:R=FAKYLWEWASVR (SEQ ID NO2); Wherein the N-end by the peptide of 3 C-ends that are modified with the peptide after the amine after 3 connect R and/or are modified with carboxymethyl by 1 is at 1 connection R '.Having only R is under the situation of a peptide, and 1 is methylated or carboxymethylation.Having only R ' is under the situation of a peptide, and 3 comprise amine.Fig. 8 has provided the embodiment of these compounds.
1,4-benzodiazepine also can connect the only part of above-mentioned peptide.For example, can connect 1,4-benzodiazepine in any position in the one or more amino acid whose above-mentioned peptides of disappearance.
When R+R '=FAKYLWEWASVR (SEQ ID NO2), its part can be by any way with 1, and 4-benzodiazepine (Bdz) connects, for example:
Bdz-AKYLWEWASVR; Or Bdz-KYLWEWASVR; Or
F-Bdz-KYLWEWASVR; Or F-Bdz-YLWEWASVR; Or
FA-Bdz-YLWEWASVR; Or FA-Bdz-LWEWASVR; Or
FAK-Bdz-LWEWASVR; Or FAK-Bdz-WEWASVR; Or
FAKY-Bdz-WEWASVR; Or FAKY-Bdz-EWASVR; Or
FAKYL-Bdz-EWASVR; Or FAKYL-Bdz-WASVR; Or
FAKYLW-Bdz-WASVR or; FAKYLW-Bdz-ASVR; Or
FAKYLWE-Bdz-ASVR; Or FAKYLWE-Bdz-SVR; Or
FAKYLWEW-Bdz-SVR; Or FAKYLWEW-Bdz-VR; Or
FAKYLWEWA-Bdz-VR; Or FAKYLWEWA-Bdz-R; Or
FAKYLWEWAS-Bdz-R; Or FAKYLWEWAS-Bdz; Or
FAKYLWEWASV-Bdz; Wherein as mentioned above, peptide is by its C-end and/or the terminal connection of N-.
Aforesaid peptide or its part also can utilize aforesaid linking group to be connected with 3 by their N-is terminal.Figure 11 has provided the embodiment of such compound.
The invention further relates to the polypeptide of forming by one of following aminoacid sequence or its part:
FAKYLW-K(bio)(SEQ?ID?NO5)
FAKYLWEW-K(bio)(SEQ?ID?NO6)
FAKYLWEWAS-K(bio)(SEQ?ID?NO7)
FAKYLWEWASVR-K(bio)(SEQ?ID?NO8)
FARFLWEWASVR-K(bio)(SEQ?ID?NO12)
FGKFLWEWASAR-K(bio)(SEQ?ID?NO13)
LGKYLWEWASAR-K(bio)(SEQ?ID?NO14)
FAKFLWEWASVR-K(bio)(SEQ?ID?NO15)
KYGW-K(bio)(SEQ?ID?NO16)
KFGW-K(bio)(SEQ?ID?NO17)
RFGW-K(bio)(SEQ?ID?NO18)
AYLW-K(bio)(SEQ?ID?NO19)
AFLW-K(bio)(SEQ?ID?NO20)
KYLW-K(bio)(SEQ?ID?NO21)
RFLW-K(bio)(SEQ?ID?NO22)
KFLW-K(bio)(SEQ?ID?NO23)
FAKYLW(SEQ?ID?NO9)
FAKYLWEW(SEQ?ID?NO10)
FAKYLWEWAS(SEQ?ID?NO11)
FARFLWEWASVR(SEQ?ID?NO24)
FGKFLWEWASAR(SEQ?ID?NO25)
LGKYLWEWASAR(SEQ?ID?NO26)
FAKFLWEWASVR(SEQ?ID?NO27)
KYGW(SEQ?ID?NO28)
KFGW(SEQ?ID?NO29)
RFGW(SEQ?ID?NO30)
AYLW(SEQ?ID?NO31)
AFLW(SEQ?ID?NO32)
KYLW(SEQ?ID?NO33)
RFLW(SEQ?ID?NO34)
KFLW(SEQ?ID?NO35)
These peptides be natural or the annexin of the HBsAg of HBV genotype A/B in conjunction with the homology variant of epi-position.Find unexpectedly that also their suppress interaction between the HBsAg of annexin V and HBV.
The polypeptide that uses in compound according to the present invention can be made up of the amino acid of L shaped formula or D form or its mixed form.In preferred embodiments, compound characteristic of the present invention is that all amino acid all is L shaped formula.In another preferred embodiment, compound characteristic of the present invention is that all amino acid all is the D form.
The present invention also comprise comprise or by polypeptide as mentioned above oppositely-compound of counter-rotating polypeptide (Guichard etc. for example, 1994 describe) composition.
Should notice that all compounds of mentioning may be lead compound, can further improve 1 on it, 4-benzodiazepine /1, the other derivatization of 4-Benzothiazepine core or peptide part.
The invention still further relates to for example composition of phosphatidylserine of polypeptide as defined above and electronegative phosphatide.Interaction among the WO97/07268 between verified phosphatidylserine and the annexin.The composition of described polypeptide and electronegative phospholipid composition can be any possible mode well known in the art, for example covalently or non-covalently the form of coupling molecule or liposome form etc.
It is above-mentioned 1 to the invention further relates to preparation, 4-benzodiazepine class or 1, the method for 4-Benzothiazepine analog derivative.Be used to prepare 1,4-benzodiazepine class or 1, the method for 4-Benzothiazepine is well known in the art.These methods include but not limited to (1963) such as Sternbach, US3109843, US3116203, US3121114, the method that US3123529 and US3203990 describe.1,4-benzodiazepine class and 1,4-Benzothiazepine class also can be buied from Sigma (Dreisenhofen, Germany) or Neosystem (Strasbourg, France) with commercial sources.From the conjugated protein polypeptides derived of annexin, more particularly, join proteic viral protein polypeptides derived from binding film, more particularly from the HBsAg polypeptides derived of HBV and/or HDV, from the Glycoprotein B of cytomegalovirus or from the conjugated protein polypeptides derived of the annexin of any influenza virus by any method well known in the art and 1,4-benzodiazepine class or 1,4-Benzothiazepine class connects.For example, by use (R, S)-Fmoc-3-amino-N-1-carboxymethyl-2-oxo-5-cyclohexyl-1,4-benzodiazepine (FB03601; N eosystem, Strasbourg, France, Fig. 7) synthesizing by the peptide based on Fmoc as structural unit can connection peptides.Fig. 8 has provided the example of the molecule that obtains.By utilize the Pierce products catalogue (Pierce, Rockford, IL, the difunctional linking group of describing in USA) of homotype makes peptide and 1 by the N-end of peptide, 3 connections of 4-benzodiazepine can be synthesized other 1,4-benzodiazepine analog derivative.Fig. 9 or 10 has provided such reaction.Figure 11 has provided the example of the molecule that obtains.
The invention still further relates to compound of the present invention prevention or treatment and wherein relate to the purposes of the medicine of annexin family member and the annexin interactional any disease between conjugated protein.
More particularly, the present invention relates to the purposes that compound prevention of the present invention or treatment wherein relate to the medicine of the interactional any virus disease between viral protein and the annexin family member.
More particularly, thus the present invention relates to compound of the present invention is disturbed the medicine of the life cycle prevention of HBV and/or HDV or treatment HBV and/or HDV infection by the interaction that suppresses HBV and/or HDV and annexin V purposes.About this point, the application of term " HBV and/or HDV " refers to that it is clearly that the HBV infection can HDV superingection this point take place separately or follow.On the other hand, HDV does not infect and takes place separately.In other words, molecule of the present invention can be used to prepare prevention or treat the medicine that independent HBV infects or blended HBV/HDV infects.
Thereby the invention further relates to the purposes that compound of the present invention is disturbed the life cycle prevention of cytomegalovirus by the interaction that suppresses cytomegalovirus and annexin II or treated the medicine of cytomegalovirus infection.
Thereby the invention further relates to the purposes that compound of the present invention is disturbed the life cycle prevention of influenza virus by the interaction that suppresses influenza virus and annexin V or treated the medicine of influenza infection.
Therefore, the present invention relates to any disease that compound prevention of the present invention or treatment wherein relate to annexin family member and protein interaction, relate more particularly to prevent or treat the interactional any virus disease that wherein relates between viral protein and the host's annexin family member, relate more particularly to prevent or treat the purposes of the medicine of HBV and/or HDV infection, cytomegalovirus infection or influenza infection.
The invention further relates to DBA753, prevent or treat any disease that wherein relates to annexin family member and protein interaction according to the compound of formula II or according to the compound of formula III, more particularly prevention or treatment wherein relate to the interactional any virus disease between viral protein and the host's annexin family member, more particularly the purposes of the medicine of prevention or treatment HBV and/or HDV infection, cytomegalovirus infection or influenza infection.
In the present invention, BDA753 is used for suppressing the interaction of HBV and annexin V especially.Although confirmed the annexin V and derived 1 of AYGW is arranged, 4-benzodiazepine class and 1, combination (the Hoffman etc. of 4-Benzothiazepine, 1998), but do not have the derived chemotactic peptide effect or 3 only derive have one amino acid whose 1,4-benzodiazepine class can not suppress the HBsAg of HBV and the interaction of annexin V.Go out that we expect be with peptide AYGW with 1,4-benzodiazepine derivatize obtains suppressing the HBsAg of HBV and the interactional compound of annexin V.
Therefore, the invention still further relates to the purposes of the peptide that comprises following aminoacid sequence or its part: AYGW (SEQ ID NO3) is used for the preparation prevention or treats wherein relating to protein and the interactional any disease of annexin family member, more particularly prevention or treatment wherein relate to the interactional any virus disease between viral protein and the host's annexin family member, more particularly the purposes of the medicine of prevention or treatment HBV and/or HDV infection, cytomegalovirus infection or influenza infection.
The invention further relates to a kind of prevention or treat and wherein relate to protein and the interactional any disease of annexin family member, more particularly prevention or treatment wherein relate to the interactional any virus disease between viral protein and the annexin family member, more particularly prevention or treatment HBV and/or HDV infect, pharmaceutical composition or medicine (two terms can exchange use) that cytomegalovirus infection or influenza infection or their any mutated viruses strain are infected, they comprise at least a compound of the present invention, at least BDA753, at least a compound according to the formula II, at least a according to the compound of formula III or peptide AYGW (SEQ ID N03) and possibly at least, a kind of pharmaceutical acceptable carrier or vehicle (two terms can exchange use).Randomly, described composition can contain other therapeutical agent that is useful on anti-above-mentioned infection.These therapeutical agents include but not limited to be used for the adefovir of HBV, BMS200475, famciclovir, phosphine formic acid, fiacitabine, fialuridine, (-)-FTC, ganciclovir, GEM132, Interferon, rabbit, lamivuding, L-FMAU, lobucavir, just-V-1326, ribavirin, Sorivudine, the compound of mentioning among vidarabine or the WO98/18818; The acyclovir that is used for cytomegalovirus, adefovir, cidofovir, ring HPMPC, fiacitabine, fomivirsen, ganciclovir, I263W94, lobucavir, ribavirin, valaciclovir or vidarabine; The amantadine that is used for influenza, GG167, GS4104, ribavirin or Rimantadine." medicine " can be used by any suitable method in technician's knowledge.Preferably, method of application can be through intestines or non-through enteral administration.Composition of the present invention can be any known drug composition forms that is fit to such medication.Described composition can be well known to a person skilled in the art method preparation, makes active compound sustained release, for example snap-out release or slowly-releasing.The active substance of these pharmaceutical compositions also can be individually dosed, do not have vehicle excipients.But they also can accept non-toxic carrier or thinner is used with pharmacy, and wherein ratio is determined by the suitability and the chemical property of concrete carrier.Known suitable carriers of technician or vehicle are salt solution, Ringer ' s solution, glucose solution, Hank ' s solution, fixed oil, ethyl oleate, 5% dextrose in saline solution, material, damping fluid and the sanitas of raising isotonicity and chemical stability.
Non-in enteral administration, medicine of the present invention with the acceptable vehicle bonded of pharmacy solution as defined above, the unitary dose injectable forms preparation of suspension or milk sap.But the dosage of administration and method will depend on individuality.Generally speaking, drug administration makes with 1 microgram/kilogram to 100 mg/kg, more preferably with 10 microgram/kilograms to 20 mg/kg, most preferably with 0.1 to 2 mg/kg to and compound of the present invention.Preferably, with the bolus dosed administration.Also can use continous pouring.If the use continous pouring can be with 1 to 100 micrograms/kg/min, more preferably the 5 dosage dabbling drugs to 20 micrograms/kg/min.
Be fit to oral dosage form and include but not limited to tablet, pill, capsule, caplets, granule, powder agent, elixir, syrup, solution and moisture or oil suspension.The per daily dose that is used for the suitable active compound of people's administration generally is about 1 milligram to about 5000 milligrams, more generally about 5 milligrams to about 1000 milligrams, and dose administration or once a day or repeatedly divided dose administration separately.
The present invention relate at last aforesaid compound screening blocking-up annexin and with the interactional any protein of annexin, more particularly, with the interactional any viral protein of annexin, more particularly, the purposes in the method for the bonded molecule of the annexin of the Glycoprotein B of the HBsAg protein of HBV, cytomegalovirus or any influenza virus between conjugated protein.As used herein, term " method of screening molecule " refers to be fit to well known in the art any method of molecular screening.Especially, this term refers to the method that embodiments of the invention 1 are described.
The present invention describes in detail by the embodiment with reference to the particularly advantageous embodiment that provides now.But should notice that these embodiment are illustrative not limiting the present invention in any way.In specification sheets and claims except as otherwise noted, term " comprises ", and the change list example is interpreted as step or the group that refers to comprise described integral part or described integral part or step as " comprising " and " containing ", but do not get rid of the step or the group of any other integral part or integral part or step.
Table and brief description of drawings
Table 1 provide relate to 6 genotype determining HBsAg (A, B, C, D, E, the polypeptide that the district of the 99-169 amino acids of " little " HBsAg F) and assert checks is in conjunction with the sequence information of annexin V.
Fig. 1 has provided 1,4-benzodiazepine class and 1, and the foundation structure formula of 4-Benzothiazepine class (formula I), wherein:
* at 1 :-Benzothiazepine class has a sulphur atom (X represents S); X also can be SO n, n=0 wherein, 1,2;
-benzodiazepine class has a nitrogen-atoms (X represents N), and it is modified with carboxymethyl (CH in addition at 1 2COOH) afterwards, allow by the C-of peptide terminal that connect or by the N-of peptide terminal connect be modified with in claims peptide or its part that is defined as " containing the peptide of a protein-bonded annexin of annexin " in addition in conjunction with epi-position.Be not modified with at N under the situation of a peptide, modification can be H, alkyl, and phenyl ,-COZ, wherein Z represents H, alkyl, the phenyl of phenyl or replacement;
* 1,2,3,4,5 can form two keys with adjoining position; If this is the case, side chain R then 2, R 4, R 5Or R 7Do not exist;
* the R on 2 and/or 2 ' 1And/or R 2Can represent H, amine, alkyl or position 2 can be oxidized (=O).At R 1And/or R 2Be under the situation of amine, it can be further or directly connects or to be connected in the N-of peptide terminal and be defined as peptide or its part of " containing the peptide of a protein-bonded annexin of annexin in conjunction with epi-position " in claims and replace by resembling the such linking group of glutaraldehyde or succinyl oxide by the C-of peptide is terminal;
* the R on 3 and/or 3 ' 3And/or R 4Can represent H, amine or alkyl.At R 3And/or R 4Be under the situation of amine, it can be further or directly connects or to be connected in the N-of peptide terminal and be defined as peptide or its part of " containing the peptide of a protein-bonded annexin of annexin in conjunction with epi-position " in claims and replace by resembling the such linking group of glutaraldehyde or succinyl oxide by the C-of peptide is terminal;
* the R on 4 5Can represent 4 to be modified with carboxymethyl (CH in addition 2COOH) back or directly terminal that connect or by being defined as the peptide of " containing the peptide of a protein-bonded annexin of annexin " or the side chain of its part in the terminal claims that connect of the N-of peptide in conjunction with epi-position by the C-of peptide.At R 5Not under the situation of the peptide that defines in claims, R 5Can represent H, alkyl ,-CO-R 12Perhaps 3-(1-(4-benzyl) piperidyl) propionyl);
* R 6And/or R 7Represent H or alkyl or phenyl, may further be substituted with alkyl, cyano group, halogen, nitro, alkyl alkoxy, alkyloyl, carboxyl, alkyloyl alkoxyl group, formamyl;
* R 8, R 9, R 10And/or R 11Represent H, alkyl, cyano group, halogen, nitro, alkyl alkoxy, alkyloyl, carboxyl, alkyloyl alkoxyl group, formamyl;
* R 12Represent H or alkyl or may further be substituted with alkyl, cyano group, halogen, nitro, alkyl alkoxy, alkyloyl, carboxyl, alkyloyl alkoxyl group, the phenyl of formamyl.
Fig. 2 has provided the structure (FR2479818) of formula II, wherein:
* at 2, the peptide that R represents an amino acid or is made up of two or three amino acid;
* X represents chlorine or fluorine.
Fig. 3 has provided the structure of formula III, wherein 1,3 of 4-benzodiazepine replaced by peptide ARPYN and 1 replaced (Nachman etc., 1998) by the L residue.
Fig. 4 has provided the structure of formula IV, wherein:
* R, R ' and/or R+R ' constitute and comprise peptide or its part of the conjugated protein annexin of annexin in conjunction with epi-position.Having only R is under the situation of a peptide, and position 1 is methylated.Having only R ' is under the situation of a peptide, and amine is contained in position 3;
* X representative can be by-COCH 3The N-end of the peptide of modifying;
* Y representative can be by-NH 2Or-the C-end of the peptide of Methionin-biotin modification.
Fig. 5 has provided the structure of formula V, and wherein peptide R connects by linking group (SPC) by its N-is terminal.
Fig. 6 provided among the embodiment 1 describe 1, the structure of 4-benzodiazepine analog derivative: BDA250:1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepine -2-ketone; BDA452:3-(R, S)-(L-tryptophyl)-1,3-hydrogen-1-methyl-5-phenyl-2H-1,4-benzodiazepine -2-ketone; BDA753:3-(R, S)-complete-L-(NH-Trp-G1y-Tyr-Ala-H)-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepine -2-ketone.
Fig. 7 has provided and has been used for 1,4-benzodiazepine analog derivative synthetic Fmoc protection (R, S)-Fmoc-3-amino-N-1-carboxymethyl-2-oxo-5-cyclohexyl-1,4-benzodiazepine (FB03601; Neosystem, Strasbourg, France).
Fig. 8 has provided some according to 1 of formula IV, the example of 4-benzodiazepine analog derivative.
Fig. 9 has provided by using glutaraldehyde to synthesize according to 1 of formula V the basic reaction of 4-benzodiazepine derivative as linking group.
Figure 10 has provided by using succinyl oxide to synthesize according to 1 of formula V the basic reaction of 4-benzodiazepine derivative as linking group.
Figure 11 has provided some according to 1 of formula V, the example of 4-benzodiazepine analog derivative.
Figure 12 proves that different compounds suppresses the interactional difference between annexin V and the HBsAg: described BDA250 in Fig. 6, BDA452 and BDA753; IGP1362 and IGP1363 are respectively from the 158-169 amino acids of the HBsAg of HBV and 115-125 amino acids deutero-peptide and provide table 1.BDA753 can suppress the interaction between annexin V and the HBsAg, and the compound that only comprises the benzodiazepine part of this molecule (BDA250) then can not.
Figure 13 proves that the different peptides of the natural variant of the 158-169 amino acids of representing HBsAg or the peptide (IGP1632) that representative C-end does not have other modified biological elementization Methionin are to interactional different suppress (tables 1) between annexin V and the HBsAg.These results are normalized to the inhibition of IGP1481 and render a service.
Figure 14 proves that the different peptides of the variant of the terminal brachymemma of C-of representing IGP1481 suppress (tables 1) to the interactional difference between annexin V and the HBsAg.These results are normalized to the inhibition of IGP1481 and render a service.
Figure 15 proves that the different peptides of the non-natural mutant of representing IGP1481 are to interactional different suppress (tables 1) between annexin V and the HBsAg.These results are normalized to the inhibition of IGP1481 and render a service.The peptide of band asterisk shows at the maximum concentration of testing does not have restraining effect, and bar is rectified and represented the highest relative concentration of test in these cases.
Embodiment
Embodiment 1:1,4-benzodiazepine and derivative are to the bonded influence of annexin V to HBsAg
In order to prove that annexin V binding molecule has carried out competition experiments to the interactional influence of HBsAg and annexin V.Annexin V binding compounds (Fig. 6 and table 1) is competed with HBsAg in conjunction with the annexin V.Measure annexin V at last with HBsAg bonded mark.In brief, recombinant HBsAg is coated on the microtiter plate (2 mcg/ml, 4 ℃ under spend the night in the TBS that is supplemented with 1mM calcium chloride and 1mM magnesium chloride).With the 3% cold dull and stereotyped back of isinglass sealing (in the TBS that is supplemented with 1mM calcium chloride and 1mM magnesium chloride, cultivating 2 hours under the room temperature), in the presence of excessive peptide or compound, add 125The annexin V (50ng) of I-mark, and make interaction (in the TBS that is supplemented with 1mM calcium chloride and 1mM magnesium chloride and 0.3% cold isinglass, cultivating 2 hours under 37 ℃).After the incubation with the TBS washing plate that is supplemented with 1mM calcium chloride and 1mM magnesium chloride and 0.05%Tween-20 three times.This test repeats 6 times, and the result of expression is mean value (line on the mean value summit is represented standard deviation).The all cpds or the peptide that use are as follows to the molar excess of HBsAg:
BDA250 25000
BDA452 13750
BDA753 1600
IGP1362 400
IGP1363 560
Can judge that from Figure 12 BDA753 and IGP1362 molecule destruction annexin V combine with HBsAg's.BDA753 has 1 of 4 amino acid whose peptide chains by deriving, 4-benzodiazepine forms.Its benzodiazepine part is identical with BDA250, and it still can not suppress the combination of HBsAg to the annexin V in conjunction with annexin.On the contrary, BDA250 has but strengthened combination.Have with the similar core texture of BDA250 but the molecule BDA452 that has an aminating tryptophane does not influence the combination of HBsAg to the annexin V, and molecule BDA753 suppresses the combination of HBsAg to the annexin V really.Can reach a conclusion from these results, BDA753 is last exist and BDA452 goes up non-existent three reasons that other amino acid is observed competition effect.Most probably these amino acid the same with HBsAg to the annexin V in the same loci combination.Already in the aminating tryptophan residue among the BDA452 also may partly disturb the HBsAg combination, because this additional tryptophan residue destroys the combination enhancing that BDA250 causes really.This shows that also can further modify BDA452 obtains better to suppress the annexin V to HBsAg bonded specificity.The peptide IGP1362 that comprises the aa158-169 of HBsAg also can suppress HBsAg to the combination (Figure 12) of annexin V and most probable combination and BDA753 in conjunction with the time same loci.Should notice that not all peptide sequence all obtains the identical result of sequence who represents with peptide IGP1363, described IGP1363 is also derived by HBsAg (amino acid/11 15-125), but does not suppress the combination of annexin V to HBsAg.The HBsAg land of the sequence debond annexin V that exists in this peptide of can reaching a conclusion.
Therefore, IGP1362 or its part connect 1,4-benzodiazepine class, BDA250 or BDA452 or its any derivative.Such molecule can use (R, S)-Fmoc-3-amino-N-1-carboxymethyl-2-oxo-5-cyclohexyl-1,4-benzodiazepine (FB03601; Neosystem, Strasbourg, France Fig. 7) synthesizes by the peptide based on Fmoc as structural unit and prepares.Fig. 8 has provided the embodiment of the molecule that obtains.By in reaction as shown in Figures 9 and 10, utilizing the linking group molecule to make peptide and 3 amine-1 by the N-end of peptide, 4-benzodiazepine connect to synthesize other 1,4-benzodiazepine analog derivative.Figure 11 has provided the embodiment of the molecule that obtains.Test obtains in competitive assay 1, and 4-benzodiazepine analog derivative proves the specificity of the raising in the interaction of blocking-up annexin V and HBsAg.
Embodiment 2: with the further evaluation of annexin V bonded HBsAg sequence
In order to study peptide sequence in more detail, further optimized in conjunction with measuring to HBsAg-annexin V bonded restraining effect.This causes following mensuration setting:
With the microtiter plate bag by the HBsAg:500ng/100 microlitre, 1 hour, 37 ℃;
Sealing: the cold isinglass of TBS3% (v/v), 1 hour, room temperature;
In conjunction with: 50ng radio-labeled annexin V/100 microlitres (at TBS, 10mM calcium chloride, in the cold isinglass of 0.3% (v/v) 37 ℃ down with the pre-cultivation of competing compound 1 hour), 1 hour, 37 ℃;
Flushing: with 200 microlitre TBS, 10mM calcium chloride, 0.05% (v/v) Tween-20, flushing is three times under the room temperature;
With the marker of SDS (60 microlitres, 1 hour, 37 ℃) elution of bound, then in gamma counter, count.
Peptide IGP1362 is the peptide that side chain is arranged, and has two side chains of amino acid/11 58-169 (table 1) of little HBsAg on biotinylated Methionin core.This sequence is also come synthetic from having the monomeric form (IGP1481 and 1623 sees Table 1) that has the terminal Methionin of C-of vitamin H or have an amidated C-end at the ε amine functional group.In addition, the same zone (amino acid/11 58-169) that HBsAg represent in preparation is still from other genotype deutero-peptide of HBV: the IGP1624 and 1625 that represents the main sequence of finding the C genotype, represent the IGP1618 and the IGP1619 that represents the main sequence of finding among the genotype F of the main sequence of finding in the D/E genotype, and IGP1481 represents the main sequence of finding in the A/B genotype (table 1).Peptide IGP1481, total sequence of 1624,1625,1618 and 1619 is represented 90% of obtainable 460 HBsAg sequences in October, the 1998 common sequence database.All peptides are all analyzed with mass spectrum the fraction that contains peptide through the reverse-phase chromatography purifying and for the existence of the peptide that confirms to expect.Calculate the concentration that causes 50% competition effect at annexin V/HBsAg in conjunction with the peptide of measuring purifying in measuring and with molar excess to the recombinant HBsAg of bag quilt.Is 17.5 to the molar excess that reaches 50% competitive level for IGP1481, and IGP1481 is and the HBsAg homologous sequence of bag quilt.The result of other peptide provides and is normalized to the value (molar excess that will reach the IGP1481 of 50% competitive level is set at 1) that obtains with IGP1481 in Figure 13.Can judge the competitive level (maximum difference is for low 5 times of IGP1625) that shows the same levels size from all peptides of other genotype deutero-from this figure.All the natural variants that can foretell the HBsAg of existence from this result show identical effectiveness.Beat all is that peptide identical with 1481 except biotinylation Methionin 1623 is active little 7 times.Therefore peptide is added that a biotinylation Methionin changes its character in such a way and makes effectiveness be enhanced.IGP1481,1618,1619,1624 or 1625 or its part connect 1,4-benzodiazepine , BDA250, BDA452 or its any derivative.In competitive assay, measure obtain 1,4-benzodiazepine analog derivative proves the specificity that they improve in the interaction of blocking-up annexin V and HBsAg.
Embodiment 3: with further describing of annexin V bonded HBsAg sequence
Important for which part of measuring sequence 158-169 for observed competition effect, synthesized the peptide of the terminal brachymemma of C-, confirm its structure by the reverse-phase chromatography purifying and by mass spectrum.Preparation is compared with IGP1481 and is lacked 2,4 and 6 amino acid whose three such peptides respectively, IGP1556,1557 and 1558 (seeing Table 1) and mensuration competition effect.The peptide IGP1556 effectiveness that only lacks 2 C-end amino acids is only little 10 times, and lacking 4 amino acid whose IGP1557 also is so, and lacks 6 amino acid whose IGP1558 active little 100 times (Figure 14).
IGP1556,1557 or 1558 or its part connect 1,4-benzodiazepine class, BDA250, BDA452 or its any derivative.In competitive assay, measure obtain 1,4-benzodiazepine analog derivative proves the specificity that they improve in the interaction of blocking-up annexin V and HBsAg.
Embodiment 4: the evaluation that has the specific compound of raising in the interaction of blocking-up annexin V and HBsAg
Two kinds of peptide IGP1623 and BDA753 suppress the interaction of annexin V and HBsAg really.And these compounds show sequence homology to a certain degree:
FAKYLWEWASVR
：：
AYGW-Bdz passes through to improve for example IGP1618 of BDA753 and IGP1623 or its natural variant, the homology between 1619,1624 and 1625, and generation has the blocking-up annexin V of raising and the specific new compound in the interaction between the HBsAg.Such compound is:
KYGW-Bdz
KFGW-Bdz
RFGW-Bdz
AYLW-Bdz
AFLW-Bdz
KYLW-Bdz
RFLW-Bdz
KFLW-BdzBdz representative wherein 3 be modified with peptide by the terminal connected amine of its C-and 1 compd B DA250 (Fig. 6) that can be modified to carboxymethyl.
Embodiment 5: for the amino acid whose evaluation in the 158-169 district of the HBsAg of the important HBV of the high specific in the interaction between blocking-up annexin V and HBsAg
Suppress residue among the aminoacid sequence 158-169 of HBsAg of the important HBV of interactional effectiveness between HBsAg and the annexin V in order to identify for peptide IGP1481, several amino acid sports L-Ala or glutamine (IGP1482,1483,1484,1485,1486,1487,1488,1489,1493 and 1622, see Table 1).All peptides are once more by the reverse-phase chromatography purifying and by mass spectrum alleged occurrence desired results.Figure 15 illustrates the relative competition of these peptides for IGP1481.These results prove that 160 amino acid is important and this lysine mutation is that L-Ala (IGP1484) is induced at least 20 times activity (the highest experimental concentration does not still constitute competition).Specify as embodiment 4, those experiments support that amino acid alanine is to the influence of the sudden change of Methionin (or arginine) among the compd B DA753.Found identical result for normally leucic 162 amino acids.This leucine sports glutamine (IGP1486) and has almost destroyed activity, and this leucine sports L-Ala (IGP1622) and causes active at least 25 times (the highest experimental concentration does not still constitute competition) of reducing.Specify as embodiment 4, those experiments support that the amino acid glycine is to the influence of leucic sudden change among the compd B DA753.Table 1:HBV genotype and detected modal sequence with annexin V bonded polypeptide
100 110 120 130 140 150 160 170geno * * * * * * * *HBsAg?A DYQGMLPVCPLIPGSTTTSTGPCKTCTTPAQGNSMFPSCCCTKPTDGNCTCIPIPSSWAFAKYLWEWASVR (SEQ?ID?NO53)HBsAg?B DYQGMLPVCPLIPGSSTTSTGPCKTCTTPAQGTSMFPSCCCTKPTDGNCTCIPIPSSWAFAKYLWEWASVR (SEQ?ID?NO54)HBsAg?C DYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGTSMFPSCCCTKPSDGNCTCIPIPSSWAFARFLWEWASVR (SEQ?ID?NO55)HBsAg?C K (SEQ?ID?NO56)HBsAg?D DYQGMLPVCPLIPGSSTTSTGPCRTCTTPAQGTSMYPSCCCTKPSDGNCTCIPIPSSWAFGKFLWEWASAR (SEQ?ID?NO57)HBsAg?E DYQGMLPVCPLIPGSSTTSTGPCRTCTTLAQGTSMYPSCCCSKPSDGNCTCIPIPSSWAFGKFLWEWASAR (SEQ?ID?NO58)HBsAg?F DYQGMLPVCPLIPGSTTTSTGPCKTCTTLAQGTSMYPSCCCSFPSDGNCTCIPIPSSWALGKYLWEWASAR (SEQ?ID?NO59)IGP1363 (TTSTGPCKTCT)2-KKGK(bio)GA (SEQ?ID?NO60)IGP1362 (FAKYLWEWASVR)2-KKGK(bio)GA (SEQ?ID?NO4)IGP1481 FAKYLWEWASVR-K(bio) (SEQ?ID?NO8)IGP1623 FAKYLWEWASVR (SEQ?ID?NO2)IGP1624 FARFLWEWASVR-K(bio) (SEQ?ID?NO12)IGP1618 FGKFLWEWASAR-K(bio) (SEQ?ID?NO13)IGP1619 LGKYLWEWASAR-K(bio) (SEQ?ID?NO14)IGP1625 FAKFLWEWASVR-K(bio) (SEQ?ID?NO15)IGP1556 FAKYLWEWAS-K(bio) (SEQ?ID?NO7)IGP1557 FAKYLWEW-K(bio) (SEQ?ID?NO6)IGP1558 FAKYLW-K(bio) (SEQ?ID?NO5)IGP1482 AAKYLWEWASVR-K(bio) (SEO?ID?NO61)IGP1483 FNKYLWEWASVR-K(bio) (SEQ?ID?NO62)IGP1484 FAAYLWEWASVR-K(bio) (SEQ?ID?NO63)IGP1485 FAKALWEWASVR-K(bio) (SEQ?ID?NO64)IGP1486 FAKYNWEWASVR-K(bio) (SEQ?ID?NO65)IGP1487 FAKYLAEWASVR-K(bio) (SEQ?ID?NO66)IGP1488 FAKYLWAWASVR-K(bio) (SEQ?ID?NO67)IGP1489 FAKYLWEAASVR-K(bio) (SEQ?ID?NO68)IGP1493 FAKYLWEWASVA-K(bio) (SEQ?ID?NO69)IGP1622 FAKYAWEWASVR-K(bio) (SEQ?ID?NO70)
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Claims (17)

1. one or more protein-bonded annexins of annexin that comprise are arranged in conjunction with the peptide of epi-position or the compound of its part according to deriving of formula I, prerequisite is that described compound is not BDA753, according to the compound of formula II or according to the compound of formula III.
2. the compound that one or two peptide R and/or R ' are arranged according to deriving of formula IV, make R, R ' and/or R+R ' representative comprises peptide or its part of the protein-bonded annexin of annexin in conjunction with epi-position, and prerequisite is that described compound is not BDA753 or according to the compound of formula III.
3. have according to deriving of formula V to comprise peptide or its part of the protein-bonded annexin of annexin in conjunction with epi-position, R, compound.
4. according to each compound of claim 1-3, wherein said annexin is conjugated protein to be viral protein.
5. according to the compound of claim 4, wherein said viral protein is the surface antigen of HBV.
6. according to the compound of claim 4, wherein said viral protein is the Glycoprotein B of cytomegalovirus.
7. according to the compound of claim 4, wherein said viral protein is from any strains of influenza viruses deutero-albumen, and condition is that this influenza proteins is in conjunction with the annexin V.
8. according to deriving of claim 4 the one or more following peptides or the compound of its part are arranged:
KTCTTPAQGN(SEQ?ID?NO1)
(FAKYLWEWASVR) 2-KKGK(bio)GA(SEQ?ID?NO4)
FAKYLW-K(bio)(SEQ?ID?NO5)
FAKYLWEW-K(bio)(SEQ?ID?NO6)
FAKYLWEWAS-K(bio)(SEQ?ID?NO7)
FAKYLWEWASVR-K(bio)(SEQ?ID?NO8)
FAKYLW(SEQ?ID?NO9)
FAKYLWEW(SEQ?ID?NO10)
FAKYLWEWAS(SEQ?ID?NO11)
FAKYLWEWASVR(SEQ?ID?NO2)
FARFLWEWASVR-K(bio)(SEQ?ID?NO12)
FGKFLWEWASAR-K(bio)(SEQ?ID?NO13)
LGKYLWEWASAR-K(bio)(SEQ?ID?NO14)
FAKFLWEWASVR-K(bio)(SEQ?ID?NO15)
KYGW-K(bio)(SEQ?ID?NO16)
KFGW-K(bio)(SEQ?ID?NO17)
RFGW-K(bio)(SEQ?ID?NO18)
AYLW-K(bio)(SEQ?ID?NO19)
AFLW-K(bio)(SEQ?ID?NO20)
KYLW-K(bio)(SEQ?ID?NO21)
RFLW-K(bio)(SEQ?ID?NO22)
KFLW-K(bio)(SEQ?ID?NO23)
FARFLWEWASVR(SEQ?ID?NO24)
FGKFLWEWASAR(SEQ?ID?NO25)
LGKYLWEWASAR(SEQ?ID?NO26)
FAKFLWEWASVR(SEQ?ID?NO27)
KYGW(SEQ?ID?NO28)
KFGW(SEQ?ID?NO29)
RFGW(SEQ?ID?NO30)
AYLW(SEQ?ID?NO31)
AFLW(SEQ?ID?NO32)
KYLW(SEQ?ID?NO33)
RFLW(SEQ?ID?NO34)
KFLW(SEQ?ID?NO35)
9. the polypeptide or its part that have one of following aminoacid sequence:
FAKYLW-K(bio)(SEQ?ID?NO5)
FAKYLWEW-K(bio)(SEQ?ID?NO6)
FAKYLWEWAS-K(bio)(SEQ?ID?NO7)
FAKYLWEWASVR-K(bio)(SEQ?ID?NO8)
FARFLWEWASVR-K(bio)(SEQ?ID?NO12)
FGKFLWEWASAR-K(bio)(SEQ?ID?NO13)
LGKYLWEWASAR-K(bio)(SEQ?ID?NO14)
FAKFLWEWASVR-K(bio)(SEQ?ID?NO15)
KYGW-K(bio)(SEQ?ID?NO16)
KFGW-K(bio)(SEQ?ID?NO17)
RFGW-K(bio)(SEQ?ID?NO18)
AYLW-K(bio)(SEQ?ID?NO19)
AFLW-K(bio)(SEQ?ID?NO20)
KYLW-K(bio)(SEQ?ID?NO21)
RFLW-K(bio)(SEQ?ID?NO22)
KFLW-K(bio)(SEQ?ID?NO23)
FAKYLW(SEQ?ID?NO9)
FAKYLWEW(SEQ?ID?NO10)
FAKYLWEWAS(SEQ?ID?NO11)
FARFLWEWASVR(SEQ?ID?NO24)
FGKFLWEWASAR(SEQ?ID?NO25)
LGKYLWEWASAR(SEQ?ID?NO26)
FAKFLWEWASVR(SEQ?ID?NO27)
KYGW(SEQ?ID?NO28)
KFGW(SEQ?ID?NO29)
RFGW(SEQ?ID?NO30)
AYLW(SEQ?ID?NO31)
AFLW(SEQ?ID?NO32)
KYLW(SEQ?ID?NO33)
RFLW(SEQ?ID?NO34)
KFLW(SEQ?ID?NO35)
10. according to each compound of claim 1-9, it comprise or by all amino acid wherein be L shaped formula or wherein all amino acid be that the polypeptide of D form is formed.
11. according to each compound of claim 1-10, it comprise or by oppositely-the counter-rotating polypeptide forms.
12. preparation is according to each the method for concrete molecule of claim 1-11.
13. each molecule preparation prevention or treatment wherein relates to protein and the interactional any disease of annexin family member according to claim 1-11, more specifically prevention or treatment wherein relate to the interactional any virus disease between viral protein and the host's annexin family member, more particularly prevention or treatment HBV and/or HDV infect the purposes of the medicine of cytomegalovirus infection or influenza infection.
14. prevent or treat wherein to relate to protein and the interactional any disease of annexin family member according to the compound of BDA753, formula II or according to the compound of formula III, more particularly prevention or treatment wherein relate to the interactional any virus disease between viral protein and the host's annexin family member, more particularly the purposes of the medicine of prevention or treatment HBV and/or HDV infection, cytomegalovirus infection or influenza infection.
15. comprising the peptide or its part: the AYGW (SEQ ID NO3) of following aminoacid sequence is used for the preparation prevention or treats wherein relating to protein and the interactional any disease of annexin family member, more particularly prevention or treatment wherein relate to the interactional any virus disease between viral protein and the host's annexin family member, more particularly the purposes of the medicine of prevention or treatment HBV and/or HDV infection, cytomegalovirus infection or influenza infection.
16. pharmaceutical composition, it contains at least a molecule according to claim 1-11 or 13-15, be used for prevention or treat wherein relating to protein and the interactional any disease of annexin family member, more particularly prevention or treatment wherein relate to the interactional any virus disease between viral protein and the host's annexin family member, more particularly prevention or treatment HBV and/or HDV infection, cytomegalovirus infection or influenza infection.
17. according to the compound of claim 1-11 or 13-15 screening blocking-up annexin and with the annexin interacting proteins, more particularly, with the interactional viral protein of annexin, more particularly, the purposes in the method for the bonded molecule of the annexin of the Glycoprotein B of the HBsAg albumen of HBV, cytomegalovirus or any influenza virus between conjugated protein.
CN99810388A 1998-09-01 1999-08-25 Benzodiazepines and benzothiazepines derivatives and HBSAG peptides binding to annexins, their compositions and use Pending CN1315963A (en)

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