CN101289500A - Long lasting fusion peptide inhibitor of viral infection - Google Patents

Abstract

Peptides with antivirus and anti-integration activity are modified to provide higher stability and improved half-life in vivo. The selected peptides include fusion inhibitors DP178 and DP107 and related peptides and analogs thereof. The modified peptides are capable of forming covalent bonds with one or more blood components, preferably a mobile blood component.

Description

The long lasting fusion peptide inhibitor of virus infection

The application's dividing an application that be the exercise question submitted on May 17th, 2000 for Chinese patent application 00807671.5 (the corresponding international application PCT/US00/13651) of " long lasting fusion peptide inhibitor of virus infection ".

Invention field

The present invention relates to modified peptide, it is the inhibitor of virus activity and/or shows anti-fusion characteristics.Particularly, the present invention relates to the modified inhibitor peptides of human immunodeficiency virus (HIV), respiratory syncytial virus (RSV), human parainfluenza virus (HPV), Measles virus (MeV) and simian immunodeficiency virus (SIV), it has long-acting in the treatment of each virus infection.The invention still further relates to the conjugate of modified peptide and endogenous carrier, particularly modified peptide and multiple mobile blood ingredient, the especially conjugate of mobile endogenous protein.

Background of invention

Though be ordinary in the normal cell biological procedures, film fusion activity also participates in various disease states, comprises that for example envelope virus enters in the cell.The known peptide class can stop or interrupt film and merge the dependency activity, comprises that for example suppressing retrovirus propagates to non-infected cells.For example, synthetic peptide DP-107 and DP-178 stride the individual domain of film (" TM ") glycoprotein gp41 derived from I type human immunodeficiency virus (" HIV-1 "), are that HIV-1 infects the effective inhibitor with HIV inductive cell-cytogamy.

Lambert etc. are at " peptide of paramyxovirus fusion (F) proteic conserved regions is effective inhibitor that virus merges " Proc.Natl.Acad.Science U.S.A., March 5,1996, Vol.93 (5), synthetic peptide DP-107 and DP-178 (T-20) are disclosed among the pp.2186-91, they stride the interior individual domain of film (TM) albumen gp41 derived from I type human immunodeficiency virus (HIV-1), are effective inhibitor that HIV-1 infects and merges.Utilize computer search strategy (computerized antiviralsearching technology, C.A.S.T.) based on the secondary structure of DP-107 and DP-178 (T20), Lambert etc. identify the DP-107 that is similar to HIV-1gp41 and conservative septuple (answering) structural domain in DP-178 district in other merges the glycoprotein of virus.Block the EC that the virus-mediated synplasm of homology forms and shows by three kinds of representative paramyxovirus respiratory syncytial virus (RSV), III type human parainfluenza virus (HPIV-3) and Measles virus (MV) deutero-antiviral peptide ₅₀Value is in 0.015-0.250 μ M scope.In addition, these peptides are very selective for the source of virus.

U.S. patent 6,013, and 263,6,017,536 and 6,020,459 are incorporated herein by reference in full at this, similarly disclose 36 amino acid whose peptide DP178, and it is equivalent to be derived from HIV-1 isolate LAI (HIV-1 _LAI) amino acid 638-673 and 38 amino acid whose peptide DP107 of gp41, it is corresponding to HIV-1 _LAIThe amino acid 558-595 of gp41, both all show the anti-HIV-1 activity.

Although disclosed many antiviral or antifusogenic peptides demonstrate effectively antiviral and/or anti-fusion-activity in the prior art, these peptides are deposited in vivo short defective plasma half-life, and this mainly removes effect of fast and peptase and proteolytic enzyme owing to serum.Therefore greatly reduced effective antiviral activity of described peptide.So, need a kind of body interior action time of more permanent method that prolongs the transformation period that has antiviral and/or antifusogenic peptides now and make these peptides.

Summary of the invention

The present invention meets the demand of these and other, and relates to the modified peptide with antiviral activity and/or anti-fusion-activity.These modified peptides provide enhanced body internal stability and have reduced susceptibility to peptase or proteasome degradation.Thereby these modified peptides have farthest reduced very frequent or even the needs of successive administration of these peptides for example.The product of different embodiments of the present invention can comprise human immunodeficiency virus (HIV), human respiratory syncytial virus (RSV), human parainfluenza virus (HPV), Measles virus (MeV) and simian immunodeficiency virus (SIV) as for example pre-and/or treatment to supporting of multiple virus infection.The modification that participates in other peptides of virus transfection (for example hepatitis, the relevant virus with other of Epstein-Barr virus) also belongs in the scope of the present invention.

The chemically reactive of peptide that the present invention relates to have antiviral and/or anti-fusion-activity modified, so these modified peptides can form stable covalent linkage with available functionality (functionalitis) reaction in the blood constitutent.In an embodiment of the invention, described modified peptide contain with blood constitutent on amino, hydroxyl or sulfydryl reaction form the reactive group of stablizing covalent linkage.In another embodiment of the present invention, reactive group can be with blood protein on the maleimide of sulfydryl reaction, described blood protein comprises fluid flow blood albumen such as albumin.

Particularly, the present invention relates to the chemical reaction sex modification of DP107 and DP178 peptide and analogue thereof, analogue comprises and contains the peptide that is derived from other (non-HIV) viral aminoacid sequences, and it is corresponding to the gp41 district of the HIV that produces DP107 and DP178 and show antiviral or anti-fusion-activity.More particularly, wherein these peptides can have antivirus action to human respiratory syncytial virus (RSV), human parainfluenza virus (HPV), Measles virus (MeV) and simian immunodeficiency virus (SIV).The invention still further relates to the chemical reaction sex modification of the peptide of SEQ ID NO:1 to SEQ ID NO:86.

The present invention also relates to be used to prevent and/or treat the composition of virus infection, wherein contain the peptide that useful described reactive group is modified with antiviral activity.More particularly, the present invention relates to be used to prevent and/or treat the composition of AIDS, human respiratory syncytial virus (RSV), human parainfluenza virus (HPV), Measles virus (MeV) and simian immunodeficiency virus (SIV).

The subordinate list summary

The present invention may be better understood by the reference subordinate list, wherein:

Table 1 has been listed common amino acid and single-letter and trigram abbreviation and protecting group commonly used.

Table 2 expression DP178 carboxyl terminal is truncate.

Table 3 expression DP178 aminoterminal is truncate.

Table 4 expression DP107 carboxyl terminal is truncate.

Table 5 expression DP107 aminoterminal is truncate.

Table 6 expression HIV-2 _NIHZDP178 analogue carboxyl terminal is truncate.

Table 7 expression HIV-2 _NIHZDP178 analogue aminoterminal is truncate.

Table 8 expression RSV F2 district DP107 analogue carboxyl terminal is truncate.

Table 9 expression RSV F2 district DP107 analogue aminoterminal is truncate.

Table 10 expression RSV F1 district DP178 analogue carboxyl terminal is truncate.

Table 11 expression RSV F1 district DP178 analogue aminoterminal is truncate.

Table 12 expression HPV3F1 district DP178 analogue carboxyl terminal is truncate.

Table 13 expression HPV3F1 district DP178 analogue aminoterminal is truncate.

Table 14 expression HPV3F1 district DP107 analogue carboxyl terminal is truncate.

Table 15 expression HPV3F1 district DP107 analogue aminoterminal is truncate.

The typical anti-RSV peptide of table 16 expression.

The typical anti-HPV3 peptide of table 17 expression.

The typical anti-SIV peptide of table 18 expression.

The typical anti-MeV peptide of table 19 expression.

The summary of sequence table

List of reference sequences will be understood the present invention better, wherein:

SEQ ID NO:1 represents the peptide sequence of DP178.

SEQ ID NO:2 represents the peptide sequence of DP107.

SEQ ID NO:3-9 represents the peptide sequence of some DP178 analogue.

SEQ ID NO:10-30 represents the peptide sequence corresponding to the RSV F1 district of DP178 and DP107 and F2 district and typical anti-RSV peptide;

SEQ ID NO:31-62 represents corresponding to the HPIV3 F1 district of DP178 and DP107 and the peptide sequence of typical anti-HPIV3 peptide;

SEQ ID NO:63-73 represents corresponding to the SIV of DP178 and the peptide sequence of typical anti-SIV peptide; And

SEQ ID NO:74-78 represents corresponding to the MeV of DP178 and the peptide sequence of typical anti-MeV peptide.

Detailed Description Of The Invention

In order to ensure understanding the present invention fully, provide following definition:

Antiviral peptide: will be meant the peptide of the virus infection that suppresses cell at this used antiviral peptide, it infects by for example suppressing cell-cytogamy or free virus.The approach that infects can comprise that film merges, as what occurred in the envelope virus situation; Or some relate to virus and cyto-architectural other fusion activities.Suppress the peptide of the virus infection of specific virus and can quote for example anti-HIV peptide, anti-RSV peptide etc. with respect to this specific virus.

Antifusogenic peptides: antifusogenic peptides is meant the capable peptide that suppresses or reduce the film fusion active level between two or more entities such as virus-cell or the cell-cell of confirmation, and it is not for there is the level that occurs the film fusion down in peptide.

HIV and anti-HIV peptide: causing the human immunodeficiency virus (HIV) of acquired immune deficiency syndrome (AIDS) is a member of the lentivirus of retrovirus.HIV exists two kinds of popular type HIV-1 and HIV-2, has identified it already and has had multiple strain separately.HIV is a target with the CD-4+ cell, and entering of virus depends on combining of HIV albumen gp41 and CD-4+ cell surface receptor.Anti-HIV peptide is meant the peptide that HIV is shown antiviral activity, comprises that suppressing free virus forms the infection of CD-4+ cell and/or the HIV inductive synplasm that suppresses to infect and do not infect between the CD-4+ cell.

SIV and anti-SIV peptide: simian immunodeficiency virus (SIV) is the slow virus that causes acquired immune deficiency syndrome (AIDS) (AIDS) sample disease in susceptible ape and monkey.Anti-SIV peptide is the peptide that SIV is shown antiviral activity, comprises the infection that suppresses the SIV pair cell and suppresses synplasm formation between infection or the non-infected cells.

RSV and anti-RSV peptide: respiratory syncytial virus (RSV) is a kind of respiratory pathogen, and especially dangerous for baby and underage child, this virus can cause bronchiolitis (inflammation of tiny air flue) and pneumonia in them.RSV is negative justice, single strand RNA virus and is the member of the Paramyxoviridae of virus.The route of infection of RSV is nose, throat, tracheae, segmental bronchus and bronchiolar mucous membrane through respiratory tract usually.Anti-RSV peptide is the peptide that RSV is shown antiviral activity, comprises that the mucomembranous cell infection and the synplasm between infection and the non-infected cells that suppress free RSV virus form.

HPV and anti-HPV peptide: human parainfluenza virus (HPIV or HPV) as RSV, is the factor that another kind causes respiratory tract disease, and as the RSV viroid, is negative justice, single strand RNA virus, and it is the member of the Paramyxoviridae of virus.There are four kinds of serotypes of having discerned HIPV--HPIV-1, HPIV-2, HPIV-3 and HPIV-4.HPIV-1 is the reason that causes croup among the children, and HPIV-1 and HPIV-2 all cause and go up lower respiratory illness.HPIV-3 is usually relevant with bronchiolitis and pneumonia.Anti-HPV peptide is the peptide that HPV is shown antiviral activity, comprises that synplasm forms between the infection that suppresses free HPV virus and infection and the non-infected cells.

MeV and anti-MeV peptide: Measles virus (VM or MeV) is a kind of the have negative justice of coating, single strand RNA virus, belongs to the Paramyxoviridae of virus.As RSV and HPV, MeV causes respiratory disease, and produce cause adding, the immunosuppression of opportunistic infection.In some cases, the MeV infection that can set up brain causes neurological (neurlogical) complication.Anti-MeV peptide is the peptide that MeV is shown antiviral activity, comprises the infection and infection and the formation of the synplasm between the infection that suppress free MeV virus.

DP-178 and DP178 analogue: unless spell out in addition or put down in writing, DP-178 is meant 36 amino acid DP-178 peptides, and it is corresponding to HIV-1 isolate LAI (HIV _LAI) gp41 glycoprotein amino-acid residue 638-673 and have sequence: YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNWF (SEQ ID NO:1) and truncate, disappearance thereof and/or insert.The truncate of DP178 peptide can comprise 3-36 amino acid whose peptide.Disappearance comprises from the DP178 peptide removes one or more amino-acid residues, and can remove a contiguous sections and a plurality of part of peptide sequence.Insertion can comprise an amino-acid residue or residue chain, and can carry out at the carboxyl of DP178 peptide or N-terminal or at the interior location of this peptide.

The DP178 peptide analogs is a peptide, and its aminoacid sequence contains except that HIV-1 _LAIOutside the sequence in peptide district of virus, it is corresponding to the gp41 district that produces DP178, with and truncate, disappearance or insertion.Described other viruses can include but not limited to other HIV isolate such as HIV-2 _NIHZ, respiratory syncytial virus (RSV), human parainfluenza virus (HPV), simian immunodeficiency virus (SIV) and Measles virus (MeV).The DP178 analogue is meant that also those pass through ALLMOTI5,107 * 178 * 4 and search for the peptide sequence with the structure similar to DP178 and/or amino acid primitive that primitive is identified or discerned with PLZIP, and it is disclosed in United States Patent (USP) 6,013,263,6,017,536 and 6,020,459 and be incorporated herein.The DP178 analogue further is meant the peptide that is called " DP178 sample ", and this term is at United States Patent (USP) 6,013, in 263,6,017,536 and 6,020,459.

DP-107 and DP107 analogue: unless spell out in addition or put down in writing, DP-107 is meant 38 amino acid whose DP-107 peptides, and it is corresponding to HIV-1 isolate LAI (HIV _LAI) gp41 glycoprotein amino-acid residue 558-595 and have sequence:

NNLLRAIEAQQHLLQLTVWQIKQLQARILAVERYLKDQ(SEQ?ID?NO：2)

And truncate, disappearance and/or insert.The truncate of DP107 peptide can contain the amino acid whose peptide of 3-38.Disappearance is to comprise from the DP107 peptide removing one or more amino-acid residues, and can remove a contiguous sections and a plurality of part of peptide sequence.Insertion can comprise an amino-acid residue or residue chain, and can carry out at the carboxyl of DP107 peptide or N-terminal or at the interior location of this peptide.

The DP107 peptide analogs is a peptide, and its aminoacid sequence contains except that HIV-1 _LAIOutside the sequence in peptide district of virus, it is corresponding to the gp41 district that produces DP107, with and truncate, disappearance or insertion.Described other viruses can include but not limited to other HIV isolate such as HIV-2 _NIHZ, respiratory syncytial virus (RSV), human parainfluenza virus (HPV), simian immunodeficiency virus (SIV) and Measles virus (MeV).The DP107 analogue is meant that also those pass through ALLMOTI5,107 * 178 * 4 and search for the peptide sequence with the structure similar to DP107 and/or amino acid primitive that primitive is identified or discerned with PLZIP, and it is disclosed in United States Patent (USP) 6,013,263,6,017,536 and 6, in 020,459.The DP107 analogue further is meant the peptide that is called " DP107 sample ", and this term is at United States Patent (USP) 6,013, definition in 263,6,017,536 and 6,020,459.

Reactive group: reactive group is the chemical group that can form covalent linkage.Described reactive group and DP-107 or DP-178 peptide or its analogue or other interested antiviral or antifusogenic peptides coupling or bonding.Reactive group is generally stable in aqueous environment; and be generally carboxyl, phosphoryl or suitable acyl group; or ester or mixed acid anhydride, or imidoether (salt), can form covalent linkage as amino, hydroxyl or sulfydryl with the functionality that is positioned at the target site place on the fluid flow blood component thus.For the overwhelming majority, described ester should comprise phenolic compound, or thiol ester, alkyl ester, phosphoric acid ester etc.

Functionality: functionality is the group that the reaction-ity group reaction in the blood constitutent and the modified antiviral peptide forms covalent linkage.Functionality comprises: the hydroxyl of the reactive entity of linked ester; Sulfydryl with maleimide, imidoether and thioester substrate reaction; The amino of bonding carboxyl, phosphoryl or acyl group; With with the carboxyl of amino bonded.

Blood constitutent: blood constitutent can be fixing or mobile.Fixed blood constitutent is non-fluid flow blood component and comprises tissue, membrane receptor, a matter albumen, scleroproein, collagen, thrombocyte, endotheliocyte, epithelial cell and relevant film and membrane receptor, somatocyte, skeletal muscle and smooth muscle cell, neurone component, osteocyte and osteoclast and whole body tissue thereof, especially relevant with circulation and lymphsystem tissue.Mobile blood constitutent is not have the fixedly blood constitutent in site in the time that prolongs arbitrarily, generally is no more than 5 minutes, and more frequent is 1 minute.These blood constitutents and film are irrelevant, and are present in for a long time in the blood and exist with the minimum concentration of at least 0.1 μ g/ml.Mobile blood constitutent comprises serum albumin, Transferrins,iron complexes, ferritin and immunoglobulin (Ig) such as IgM and IgG.The transformation period of mobile blood constitutent was at least about 12 hours.

Protecting group: protecting group is to be used for protecting peptide derivant that the chemical group of id reaction does not take place.The kinds of protect base is disclosed in this paper and U.S.5, in 493,007 (being hereby incorporated by).This type of protecting group comprises ethanoyl, fluorenyl methoxy carbonyl (Fmoc), tert-butoxycarbonyl (Boc), benzyloxycarbonyl (CBZ) etc.Concrete protection amino acid is listed in the table 1.

Linking group: connect (at interval) group and be reactive entity bonding or be connected in the chemical group of antiviral or antifusogenic peptides.Linking group can comprise one or more moieties, alkoxyl group part, alkenyl part, alkynyl part or the amino part that is replaced by moieties, cycloalkyl moiety, many loop sections, aryl moiety, polyaryl part, substituted aryl part, heterocyclic moiety and substituted heterocycle part; Linking group also can contain polyethoxye amino acid, for example AEA ((2-amino) ethoxyacetic acid) or preferred linking group AEEA ([2-(2-amino) oxyethyl group)] ethoxyacetic acid).

Responsive functional group-responsive functional group is a group of representing the potential reaction site on antiviral and/or the antifusogenic peptides.If exist, the tie point that can select responsive functional group to modify as linker-reactive group.Responsive functional group includes but not limited to carboxyl, amino, sulfydryl and hydroxyl.

Modified peptide-modified peptide is the antiviral and/or antifusogenic peptides by the ligation base group modification.Described reactive group can be connected with described peptide by linking group, or does not randomly adopt linking group.Also can consider one or more additional amino acid are introduced described peptide so that the connection of reactive group.Modified peptide can vivo medicine-feeding makes it take place in vivo and the combining of blood constitutent, and they is combined and with coupling peptide (as giving a definition) vivo medicine-feeding of gained with the extracorporeal blood component.

The coupling peptide-coupling peptide is and reactive group and the covalent linkage functionality of blood constitutent between, under dispensable linking group link coupled modified peptide of blood constitutent by modified peptide.Term " coupling peptide " used among the application can more specifically refer to specific coupling peptide, for example " coupling DP178 " or " coupling DP107 ".

In view of above-mentioned definition, the present invention has the superiority of the characteristic that has antiviral and antifusogenic peptides now.The virus that can utilize described peptide to suppress for example includes but not limited at United States Patent (USP) 6,013,263,6,017,536 and 6,020, and institute's toxic strain of institute's influenza virus among 459 Table V-VII and the IX-XIV.These viruses comprise that for example: human inverse transcription virus comprises HIV-1, HIV-2; And human T lymphotropic virus (HTLV-I and HTLV-II); With inhuman retrovirus, comprise bovine leukosis's virus, cat sarcoma virus, feline leukaemia virus, simian immunodeficiency virus (SIV), simian sarcoma virus, ape and monkey leukemia and zwogerziekte virus.Non-retrovirus also can suppress by peptide of the present invention, comprises human respiratory syncytial virus (RSV), canine distemper virus, Avian pneumo-encephalitis virus, human parainfluenza virus (HPIV), influenza virus, Measles virus (MeV), dust-Pasteur's virus, hepatitis B virus and ape and monkey Mason-Pfizer virus.Non-envelope virus also can suppress with peptide of the present invention, and includes but not limited to picornavirus such as poliovirus, hepatitis A virus, enterovirus, Echo virus, Coxsackie virus, papova viruses such as papillomavirus, parvovirus, adenovirus and reovirus.

As described in embodiment, the mechanism of action of HIV fusogenic peptide is inquired in the application's background parts, and the antiviral and anti-fusion characteristics of described peptide was established already well.Demonstrated the virus-mediated cell-cytogamy of inhibition under low-down concentration corresponding to the synthetic peptide of C-terminal ectodomain sequence (for example, the amino-acid residue 643-678 of the Type B HIV-1 of LAI strain or be derived from the residue 638-673 and the residue 558-595 of similar strain).The leucine zipper structure district competition of fusogenic peptide and natural viral gp41, viral interference enters the fusion/infection of cell thus.

Of the present inventionly focus on utilizing DAC (pharmaceutical activity mixture) technology, modify selected antiviral and/or antifusogenic peptides on the protein carrier by optionally peptide being coupled at, give the application of the improved bioavailability of this peptide, prolongation and better distribution, but do not change the ntiviral characteristic of peptide.It is albumin that the present invention selects the carrier of (but being not limited to), by its free sulfhydryl groups and the antiviral and/or antifusogenic peptides coupling of modifying with maleimide base group.

Open already in the literature to the very effective some peptide sequences of the prevention of HIV-1 fusion/infection.For example, peptide DP178 is in conjunction with the conformation of the gp41 relevant with fusion.Therefore in an embodiment of the invention, modify DP178 and DP178 sample peptide.Similarly, other embodiments of the present invention comprise DP107 and the DP107 sample peptide that is used for anti-HIV and are similar to the DP107 that finds in RSV, HPV, MeV and SIV virus and the modification of the peptide of DP178.

1. DP178 and DP107

A. The DP178 peptide

The DP178 peptide is corresponding to being derived from HIV-1 _LAIThe amino-acid residue 638-673 of the transmembrane protein gp41 of isolate, and have 36 amino acid whose sequences (reading to C-terminal) from amino:

NH ₂-YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNWF-COOH(SEQ?ID?NO：1)

Except 36 aggressiveness of total length DP178, peptide of the present invention comprises truncate (that is, the magnitude range of peptide is the poly-polypeptide of tripeptides to 36) of the DP178 peptide that contains 3-36 amino-acid residue, and these truncate peptides are shown in table 2 and 3.

The amino acid of DP178 is replaced and is also belonged in the scope of the present invention in addition.HIV-1 is different on structural formula with the HIV-2 envelope protein, but at the region memory of HIV-1 and the corresponding DP178 of HIV-2 in tangible amino acid conservative property.This amino acid conservative property has periodically, has pointed out the conservative property of some structures and/or function.What therefore, amino acid was replaced a kind ofly may type will comprise that those expectations stablize the amino acid of the structure of DP178 peptide of the present invention and change.Utilize DP178 described herein and DP178 analogue sequence, one skilled in the art can compile the DP178 consensus sequence at an easy rate and determine to represent the conservative amino acid residue of preferred amino acid replacement thus.

Amino acid is replaced can have conservative or non-conservation.Conservative amino acid is replaced by forming with one or more amino acid of the amino-acid substitution DP178 peptide sequence with similar electric charge, size and/or hydrophobic property, and for example L-glutamic acid (E) is replaced the amino acid of aspartic acid (D).Non-conservation is replaced by forming with one or more amino acid of the amino-acid substitution DP178 peptide sequence with dissimilar charges, size and/or hydrophobic property, and for example L-glutamic acid (E) is to the replacement of Xie Ansuan (V).

The aminoacid insertion of DP178 can be made up of an amino-acid residue or residue chain.Insertion can be carried out at the carboxyl or the N-terminal of DP178 or the truncate peptide of DP178, and can carry out at the interior location of peptide.

Described being inserted in is generally 2-15 amino acid on the length.Considerablely be, can be in the magnitude range of more widening in the insertion that the carboxyl or the N-terminal of peptide interested carries out, preferred about 2 to about 50 amino acid.One or more described insertions can be introduced DP178 or DP178 truncate in, as long as the peptide that such insertion makes gained still can be by above-mentioned 107 * 178 * 4, ALLMOTI5 or the identification of PLZIP search primitive.

It is length range at about 2 peptides to about 50 amino-acid residues that preferred amino or C-terminal insert, and these peptides are respectively corresponding to the gp41 albumen zone of actual DP178 gp41 aminoacid sequence amino or carboxyl side.So preferred N-terminal or C-terminal aminoacid insertion will contain the gp41 aminoacid sequence, this sequence is next to the amino or the carboxyl terminal in the proteic DP178 of gp41 district.

The truncate disappearance of DP178 or DP178 also belongs in the scope of the present invention.Described disappearance is made up of one or more amino acid of removing DP178 or DP178 sample peptide sequence, and the length of gained peptide sequence following is limited to 4-6 amino acid.

Described disappearance can comprise a contiguous sections or the more than one discontinuous part of peptide sequence.One or more such disappearances can be introduced DP178 or DP178 truncate in, as long as the peptide that these disappearances obtain can be by above-mentioned 107 * 178 * 4, ALLMOTI5 or the identification of PLZIP search primitive.

B. The DP107 peptide

DP107 is a kind of 38 amino acid whose peptides, and it has effective antiviral activity, and corresponding to being derived from HIV-1 _LAIThe amino-acid residue 558-595 of the transmembrane protein gp41 of isolate, as follows:

NH ₂-NNLLRAIEAQQHLLQLTVWQIKQLQARILAVERYLKDQ-COOH(SEQ?ID?NO：2)

Except 38 aggressiveness of total length DP107, peptide of the present invention comprises truncate (that is, the magnitude range of peptide is the poly-polypeptide of tripeptides to 38) of the DP107 peptide that contains 3-38 amino-acid residue, and these truncate peptides are shown in table 4 and 5.

The amino acid of DP178 is replaced and is also belonged in the scope of the present invention in addition.As DP178, in the zone of HIV-1 and the corresponding DP178 of HIV-2, also there is tangible amino acid conservative property.This amino acid conservative property has periodically, has pointed out the conservative property of structure and/or function.What therefore, amino acid was replaced a kind ofly may type comprises that those expectations stablize the amino acid of the structure of DP107 peptide of the present invention and change.Utilize DP107 described herein and DP107 analogue sequence, one skilled in the art can compile the DP107 consensus sequence at an easy rate and determine to represent the conservative amino acid residue of preferred amino acid replacement thus.

Amino acid is replaced can have conservative or non-conservation.Conservative amino acid is replaced by forming with one or more amino acid of the amino-acid substitution DP107 peptide sequence with similar electric charge, size and/or hydrophobic property, and for example L-glutamic acid (E) is replaced the amino acid of aspartic acid (D).Non-conservation is replaced by forming with one or more amino acid of the amino-acid substitution DP107 peptide sequence with dissimilar charges, size and/or hydrophobic property, and for example L-glutamic acid (E) is to the replacement of Xie Ansuan (V).

Aminoacid insertion can be made up of an amino-acid residue or residue chain.Insertion can be carried out at the carboxyl or the N-terminal of DP107 or the truncate peptide of DP107, and can carry out at the interior location of peptide.

Described being inserted in is generally 2-15 amino acid on the length.Considerablely be, can be in the magnitude range of more widening in the insertion that the carboxyl or the N-terminal of peptide interested carries out, preferred about 2 to about 50 amino acid.One or more described insertions can be introduced DP107 or DP107 truncate in, as long as the peptide that such insertion obtains still can be by above-mentioned 107 * 178 * 4, ALLMOTI5 or the identification of PLZIP search primitive.

It is length range at about 2 peptides to about 50 amino-acid residues that preferred amino or C-terminal insert, respectively corresponding to the gp41 albumen zone of actual DP107 gp41 aminoacid sequence amino or carboxyl side.So preferred N-terminal or C-terminal aminoacid insertion will contain the gp41 aminoacid sequence, this sequence is next to the amino or the carboxyl terminal in the proteic DP107 of gp41 district.

The truncate disappearance of DP107 or DP107 also belongs in the scope of the present invention.Described disappearance comprises one or more amino acid of removing DP107 or DP107 sample peptide sequence, and the length of gained peptide sequence following is limited to 4-6 amino acid.

Described disappearance can comprise a contiguous sections or the more than one discontinuous part of peptide sequence.One or more such disappearances can be introduced DP107 or DP107 truncate in, as long as the peptide that these disappearances obtain can be by above-mentioned 107 * 178 * 4, ALLMOTI5 or the identification of PLZIP search primitive.

The truncate United States Patent (USP) 5,656 that comprehensively is disclosed in of DP107 and DP107, in 480, it is incorporated herein by reference in full at this.

2. DP107 and DP178 analogue

Analogue corresponding to above-mentioned DP178 of the present invention, DP178 are truncate, the peptide of DP107 and the truncate sequence of DP107 can be found in other virus, comprise for example non-HIV-1 envelope virus, non-envelope virus and other non-viral organism.

Described DP178 and DP107 analogue can be for example corresponding to the peptide sequence in film (" the TM ") albumen of striding that is present in envelope virus, and can be corresponding to the peptide sequence that is present in non-coating and the non-viral organism.Described peptide can have anti-fusion-activity, antiviral activity, most particularly to finding that wherein the virus of native sequences has specific antiviral activity, perhaps can show the performance of regulating the born of the same parents' internal procedure that relates to the coiled coil structure.

A. The DP178 analogue

The DP178 analogue is a peptide, and its aminoacid sequence contains for example aminoacid sequence in the peptide district of other (being non-HIV-1) virus, and this aminoacid sequence is corresponding to the gp41 peptide district that produces DP178.Described virus can include but not limited to other HIV-1 isolate and HIV-2 isolate.

DP178 analogue derived from the corresponding gp41 peptide district of other (being non-HIV-1LAI) HIV-1 isolate can comprise peptide sequence for example as follows:

NH2-YTNTIYTLLEESQNQQEKNEQELLELDKWASLWNWF-COOH(SEQ?ID?NO：3)

NH2-YTGIIYNLLEESQNQQEKNEQELLELDKWANLWNWF-COOH(SEQ?ID?NO：4)

NH2-YTSLIYSLLEKSQIQQEKNEQELLELDKWASLWNWF-COOH(SEQ?ID?NO：5)

The peptide of SEQ ID NO:3, SEQ ID NO:4 and SEQ ID NO:5 is respectively derived from HIV-1 _SF2, HIV-1 _RFAnd HIV-1 _MNOther DP178 analogue comprises derived from those of HIV-2, comprises the peptide of SEQ ID NO:6 and SEQ ID NO:7, and they are respectively derived from HIV-2 _RODAnd HIV-2 _NIHZOther effective analogue comprises the peptide of SEQ ID NO:8 and SEQ IDNO:9, verified they have antiviral activity.

In the present invention, preferred DP178 analogue is represented the peptide of those its aminoacid sequences corresponding to the proteic DP178 of gp41 district; In addition, can consider that also this peptide described herein contains length range at about 2 aminoacid sequences to about 50 amino-acid residues, this sequence is corresponding to the gp41 district of actual DP178 aminoacid sequence amino or carboxyl terminal.

Table 6 and table 7 expression HIV-2 _NIHZPossible truncate of some of DP178 analogue, they contain the peptide (being that the peptide magnitude range is the poly-polypeptide of tripeptides to 36) of 3 to 36 amino-acid residues.Peptide sequence in these tables is to list to carboxyl (right-hand) end from amino (left).

B. Other DP178 analogue and DP107 analogue

DP178 and DP107 analogue are by for example using one or more above-mentioned 107 * 178 * 4, ALLMOTI5 or PLZIP computer aided search strategy to discern and identify.These search strategies identify precognition and have the structure that is similar to DP107 and/or DP178 and/or the additional peptide district of aminoacid sequence feature.

Above-mentioned search strategy is disclosed in United States Patent (USP) 6,013 comprehensively, and 263,6,017,536 and 6,020, among the embodiment in 459 the 9th part.Though this search strategy part is based on the primary amino acid primitive of being deduced by DP107 and DP178, but it is not only based on search primary amino acid sequence homology, because this protein sequence homology is present in the main cohort of virus, but is not present between each main cohort.For example, the primary amino acid sequence homology is in the TM of the different strains of HIV-1 albumen or very high in the TM albumen of the different isolates of simian immunodeficiency virus (SIV).

Be disclosed in United States Patent (USP) 6,013, the computer search strategy in 263,6,017,536 and 6,020,459 has successfully been differentiated the proteic zone that is similar to DP107 or DP178.This search strategy is designed to use with commercially available sequence library bag, preferred PC/Gene.

At United States Patent (USP) 6,013, in 263,6,017,536 and 6,020,459, designed a series of search primitive 107 * 178 * 4, ALLMOTI5 and PLZIP primitive, and through engineering approaches in arriving wide strict scope as strict as possible, wherein preferred 107 * 178 * 4.Sequence through above-mentioned search primitive evaluation, for example those are at United States Patent (USP) 6,013,263,6,017,536 and 6,020, the listed sequence that is incorporated herein by reference with the application among Table V-XIV in 459 has anti-(as the antiviral) activity that merges potentially, can be used for anti-evaluation of merging (as antiviral) compound in addition.

3. Other antiviral peptide

A. Anti-RSV peptide

Anti-RSV peptide comprises the DP178 and/or the DP107 analogue that are gone out by corresponding peptides Sequence Identification among the RSV, and its further evaluation can suppress the virus infection of RSV.Interested this type of peptide comprises the peptide of table 16 and the peptide of SEQ ID NO:10 to SEQ ID NO:30.Interested especially is following peptide:

YTSVITIELSNIKENKCNGAKVKLIKQELDKYK(SEQ?ID?NO：14)

TSVITIELSNIKENKCNGAKVKLIKQELDKYKN(SEQ?ID?NO：15)

VITIELSNIKENKCNGAKVKLIKQELDKYKNAV(SEQ?ID?NO：16)

IALLSTNKAVVSLSNGVSVLTSKVLDLKNYIDK(SEQ?ID?NO：29)

The peptide of SEQ ID NO:10 is derived from the F2 district of RSV, and at United States Patent (USP) 6,103, utilizes described search primitive (i.e. " DP107/178 the sample ") evaluation corresponding to DP107 and DP178 peptide in 236 and 6,020,459.The peptide of SEQ ID NO:14 to SEQ ID NO:16 has contained aminoacid sequence among the SEQ ID NO:10 respectively, and demonstrated anti-RSV activity separately, particularly formed at the fusion and the synplasm that suppress rsv infection under the concentration that is lower than 50 μ g/m1 and do not infect between the Hep-2 cell.

The peptide of SEQ ID NO:11 is derived from the F1 district of RSV, and at United States Patent (USP) 6,103, utilizes described search primitive (i.e. " DP107 the sample ") evaluation corresponding to DP107 in 236 and 6,020,459.SEQ ID NO:29 contains contained aminoacid sequence among the SEQ ID NO:10, and similarly shows anti-RSV activity, particularly forms at the fusion and the synplasm that suppress rsv infection under the concentration that is lower than 50 μ g/ml and do not infect between the Hep-2 cell.

B. Anti-HPIV peptide

Anti-HPIV peptide comprises by the DP178 of corresponding peptides Sequence Identification among the HPIV and/or DP107 analogue, and it further is accredited as the virus infection that can suppress HPIV.Interested this type of peptide comprises the peptide of table 17 and the peptide of SEQ ID NO:31 to SEQ ID NO:62.Interested especially is following peptide:

VEAKQARSDIEKLKEAIRDTNKAVQSVQSSIGNLI(SEQ?ID?NO：52)

RSDIEKLKEAIRDTNKAVQSVQSSIGNLIVAIKSV(SEQ?ID?NO：58)

NSVALDPIDISIELNKAKSDLEESKEWIRRSNQKL(SEQ?ID?NO：35)

ALDPIDISIELNKAKSDLEESKEWIRRSNQKLDSI(SEQ?ID?NO：38)

LDPIDISIELNKAKSDLEESKEWIRRSNQKLDSIG(SEQ?ID?NO：39)

DPIDISIELNKAKSDLEESKEWIRRSNQKLDSIGN(SEQ?ID?NO：40)

PIDISIELNKAKSDLEESKEWIRRSNQKLDSIGNW(SEQ?ID?NO：41)

IDISIELNKAKSDLEESKEWIRRSNQKLDSIGNWH(SEQ?ID?NO：42)

The peptide of SEQ ID NO:31 is derived from the F1 district of HPIV-3, and at United States Patent (USP) 6,103, utilizes the described search primitive (i.e. " DP107 sample ") of corresponding DP107 to identify in 236 and 6,020,459.The peptide of SEQ ID NO:52 and SEQ ID NO:58 has contained aminoacid sequence among the SEQ ID NO:30 respectively, and demonstrate anti-HPIV-3 activity separately, particularly fusion between the CV-1W cell of Hep-2 cell that suppresses the HPIV-3-infection under the concentration that is lower than 1 μ g/ml and not infection and synplasm form.

The peptide of SEQ ID NO:32 is derived from the F1 district of HPIV-3, and at United States Patent (USP) 6,103, utilizes the described search primitive (i.e. " DP178 sample ") of corresponding DP178 to identify in 236 and 6,020,459.The peptide of SEQ ID NO:35 and SEQ ID NO:38 to SEQ IN NO:42 has contained aminoacid sequence among the SEQ ID NO:32 respectively, and demonstrate anti-HPIV-3 activity separately, particularly fusion between the CV-1W cell of Hep-2 cell that suppresses the HPIV-3-infection under the concentration that is lower than 1 μ g/ml and not infection and synplasm form.

C. Anti-MeV peptide

Anti-MeV peptide is the DP178 and/or the DP107 analogue that are gone out by corresponding peptides Sequence Identification in the Measles virus (MeV), and it further is accredited as the viral infection that can suppress Measles virus.Interested especially this type of peptide comprises the peptide of table 19 and the peptide of SEQ ID NO:74 to SEQ ID NO:86.Interested especially is following peptide:

HRIDLGPPISLERLDVGTNLGNAIAKLEAKELLE(SEQ?ID?NO：77)

IDLGPPISLERLDVGTNLGNAIAKLEAKELLESS(SEQ?ID?NO：79)

LGPPISLERLDVGTNLGNAIAKLEAKELLESSDQ(SEQ?ID?NO：81)

PISLERLDVGTNLGNAIAKLEAKELLESSDQILR(SEQ?ID?NO：84)

At United States Patent (USP) 6,103, utilize the described search primitive (i.e. " DP178 sample ") of corresponding DP178 to identify in 236 and 6,020,459 derived from the sequence of Measles virus.The peptide of SEQ ID NO:77, SEQ ID NO:79, SEQ ID NO:81 and SEQ ID NO:83 has the aminoacid sequence of identifying thus respectively, and demonstrate anti-MeV activity separately, particularly fusion between the Vero cell of Hep2 cell that suppresses the MeV infection under the concentration that is lower than 1 μ g/ml and not infection and synplasm form.

D. Anti-SIV peptide

Anti-SIV peptide is the DP178 and/or the DP107 analogue that are gone out by corresponding peptides Sequence Identification among the SIV, and it further is accredited as the viral infection that can suppress SIV.Interested this type of peptide comprises the peptide of table 18 and the peptide of SEQ ID NO:63 to SEQ ID NO:73.Interested especially is following peptide:

WQEWERKVDFLEENITALLEEAQIQQEKNMYELQK(SEQ?ID?NO：64)

QEWERKVDFLEENITALLEEAQIQQEKNMYELQKL(SEQ?ID?NO：65)

EWERKVDFLEENITALLEEAQIQQEKNMYELQKLN(SEQ?ID?NO：66)

WERKVDFLEENITALLEEAQIQQEKNMYELQKLNS(SEQ?ID?NO：67)

ERKVDFLEENITALLEEAQIQQEKNMYELQKLNSW(SEQ?ID?NO：68)

RKVDFLEENITALLEEAQIQQEKNMYELQKLNSWD(SEQ?ID?NO：69)

KVDFLEENITALLEEAQIQQEKNMYELQKLNSWDV(SEQ?ID?NO：70)

VDFLEENITALLEEAQIQQEKNMYELQKLNSWDVF(SEQ?ID?NO：71)

DFLEENITALLEEAQIQQEKNMYELQKLNSWDVFG(SEQ?ID?NO：72)

FLEENITALLEEAQIQQEKNMYELQKLNSWDVFGN(SEQ?ID?NO：73)

The sequence of striding synexin derived from SIV is at United States Patent (USP) 6,103, utilizes the described search sequence (i.e. " DP178 sample ") of corresponding DP178 to identify in 236 and 6,020,459.The peptide of SEQ ID NO:64 to SEQ ID NO:73 has the aminoacid sequence of identifying thus respectively, and demonstrates effective anti-SIV activity as the crude product peptide separately.

4. The modification of antiviral and antifusogenic peptides

The present invention relates to have the modification of the peptide of antiviral and/or anti-fusion-activity, comprise this modification of DP-107 and DP-178 and analogue thereof.Described modified peptide can react through covalent linkage with the reactive functionalities utilized in the blood constitutent.The invention still further relates to described modification, with the combining and application method of blood constitutent.These methods comprise and use coupling peptide not to the patient and compare the effective treatment life-span that prolongs coupling antiviral peptide derivative.Described modified peptide is the peptide that is designed to DAC (the affine mixture of medicine) type, its contain antiviral peptide molecule and linking group and can with the reactive group of the reactive functionalities reaction of mobile blood protein.By with blood constitutent or albumen test, described modified peptide or DAC can be passed to suitable site or acceptor through blood.

For with albumen on functionality form covalent linkage, people can utilize various active carboxyl, particularly ester as reactive group, wherein hydroxyl is that physiology is acceptable in the needed level of modified peptide.When adopting multiple different hydroxyl in these reactive groups, modal will be N-hydroxy-succinamide or (NHS), N-hydroxyl-sulfosuccinimide (sulfo group-NHS).In preferred implementation of the present invention, functionality on the protein is that sulfydryl and reactive group are the groups that contains dimaleoyl imino, and for example γ-maleimide-butyramide (butyralamide) (GMBA) and dimaleoyl imino (maleimido) propionic acid (MPA).

Primary amine is the main target of NHS ester.Available alpha-amino group that exists on the proteinic N-terminal and the reaction of NHS ester.Yet the alpha-amino group on the protein also can not be that the NHS coupling is needed or available.When 5 amino acid are nitrogenous in its side chain, have only the ε-amine of Methionin to react with the NHS ester significantly.When discharging N-hydroxy-succinamide, the linked reaction of NHS ester and primary amine can form amido linkage, shown in following route

In preferred implementation of the present invention, the functional group on this albumen should be that sulfydryl and reactive group chemically should be the groups that contains dimaleoyl imino, as MPA and GMBA (γ-maleimide-butyramide).When the pH of reaction mixture maintains sulfydryl on the first-selected peptide of 6.5 dimaleoyl iminos between 7.4 time.PH 7.0 times, fast 1000 times of the speed of the speed ratio of the reaction of dimaleoyl imino and sulfydryl and amine reaction.Form stable thioether bond between dimaleoyl imino and the sulfydryl, this thioether bond can not rupture under physiological condition, shown in following route.

A. specific marker

Preferably, modified peptide of the present invention be designed to specifically with fluid flow blood protein on the sulfydryl reaction.This type of reaction is suitable to setting up with the peptide of maleimide key (for example, by GMBS, MPA or other maleimide) modification and the covalent linkage between the sulfydryl on the fluid flow blood albumen (as serum albumin or IgG).

Under some environment, adopt the specific marker of maleimide to provide than some kinds of better superiority of mobile proteic non-specific mark with group such as NHS and sulfo group-NHS.Sulfydryl is abundant not as amino acid in vivo.So, the amine-modified peptide (being the maleimide peptide) of maleimide of the present invention should with less albumen covalent bonding.For example, in albumin (the abundantest blood protein), only there is a sulfydryl.Therefore, peptide-maleimide-albumin conjugate will tend to contain about 1: 1 peptide of mol ratio and albumin.Except that albumin, IgG molecule (II type) also has free sulfhydryl groups.Because IgG molecule and serum albumin have constituted the overwhelming majority of soluble proteins in the blood, thereby they also constitute most covalently bound free sulfhydryl groups of peptide that can be amine-modified with maleimide in the blood.

In addition, in containing the blood protein of free sulfhydryl groups (comprising the IgG class), cause the preferential formation of peptide-maleimide-albumin conjugate with the maleimide specific marker, this is owing to the unique property of albumin self.The albuminous unique free sulfhydryl groups conservative at multiple material camber is positioned at amino-acid residue 34 (Cys ³⁴) on.Confirmed albuminous Cys at present already ³⁴Has higher reactivity than the free sulfhydryl groups that is positioned on other protein that contain free sulfhydryl groups.This part is owing to albumin Cys ³⁴Has low-down pK value 5.5.This value is more much lower than the typical pK value of common cysteine residues, and described typical pK value is generally about 8.Since this low pK, albuminous Cys under normal physiological condition ³⁴Mainly be the ionization form, this has greatly improved its reactivity.Except low Cys ³⁴Beyond the pK value, another improves Cys ³⁴Reactive factor be its position, it is positioned at the crack on the surface of a ring texture that approaches albumin V district.Such position makes Cys ³⁴Be very easy to by the part utilization of all kinds, and at Cys ³⁴It in the biological action as radical ravin and free sulfhydryl groups scavenging agent an important factors.These characteristics make Cys ³⁴Be very easy to and maleimide-reactive polypeptide, and the quickening of speed of reaction can be 1000 times that maleimide-peptide and other contain the proteic speed of reaction of free sulfhydryl groups.

Another superiority of peptide-maleimide-albumin conjugate is a circulation ratio, this circulation ratio with at Cys ³⁴Locating 1: 1 specific peptide loads on the albumin relevant.The selectivity of other technologies as glutaraldehyde, DCC, EDC and other chemical activations (as unhindered amina) lack.For example, albumin contains 52 lysine residues, and 25-30 wherein is positioned at albuminous surface and is also therefore utilized by linked reaction.Activate these lysine residues, perhaps in addition peptide is modified with by these lysine residue couplings, obtain allos conjugate colony.Even adopt the peptide and the albumin of 1: 1 mol ratio, product will be made up of multiple coupled product, some contain 0,1,2 or a polypeptide/albumin more, and have at random link coupled peptide on any one or a plurality of site in 25-30 available lysine site separately.Owing to may there be multiple array configuration, the difficulty so the characterization of precise combination thing and each coupling batch character becomes, and batch and batch between circulation ratio almost be impossible, cause this type of conjugate can't become desired therapeutical agent.In addition, though as if should possess the advantage of the more therapeutical agents of each albumin molecule transmission at least, studies show that 1: 1 therapeutical agent and albumin ratio is more desirable by the coupling of albuminous lysine residue." the cancer target characteristic of load factor decision methotrexate-albumin conjugate in rat " at Stehle etc. Anti-Cancer Drugs, Vol.8, among the pp.677-685 (1997) (document is hereby incorporated by), the author reports anticancer methotrexate and produces most promising result through albuminous 1: 1 ratio of glutaraldehyde link coupled.These conjugates are preferentially absorbed by tumour cell, and the conjugate that carries 5: 1 to 20: 1 methotrexate molecules has changed HPLC and distributes and absorbed by liver rapidly in vivo.Suppose thus that under these higher ratios albuminous conformational change has weakened its validity as the treatment carrier.

By the vivo medicine-feeding of control maleimide-peptide, people also can control volume in the specific marker of albumin and IgG.In the administration of routine, the 80-90% of the maleimide-peptide of using is with tagged albumin and less than 5% mark IgG.The trace mark that free sulfhydryl groups such as gsh also can occur.This type of specific marker is suitable for using in the body, the theoretical transformation period that it can medicine that accurate calculation is given.

Except controlled body internal specific mark was provided, maleimide-peptide can provide the specific marker of exsomatize serum albumin and IgG.This in vitro marker comprise maleimide-peptide added blood, serum or contain serum albumin and/or the salt brine solution of IgG in.In case occur and the coupling of exsomatizing of maleimide-peptide, blood, serum or salt brine solution can be administered to once more and be used for interior therapeutic in the blood samples of patients.

With the contrast of NHS-peptide, maleimide-peptide is general quite stable in the presence of the aqueous solution and unhindered amina.Because maleimide-peptide only reacts with free sulfhydryl groups, does not need usually to prevent maleimide-peptide generation id reaction with protecting group.In addition, the enhancing of modified peptide stability allow to use further purification step, for example utilize the product that uses in the very pure suitable body of HPLC preparation.At last, the chemical stability that has strengthened provides the longer transformation period for product.

B. non-spy's property led mark

Antiviral peptide of the present invention also can be used for the non-specific mark of blood constitutent by modification.For non-specific mark, also adopt and amino bonded, especially form amido linkage.In order to form this key, can adopt various active carboxyl, particularly ester as chemically reactive group, wherein hydroxylic moiety is physiologically acceptable under desired level.Though can adopt multiple different hydroxyl in these linking agents, and the most frequently used is N-hydroxy-succinamide (NHS) and N-hydroxyl-sulfosuccinimide (sulfo group-NHS).

Other available linking agents are disclosed in the U.S. patent 5,612,034, and it is hereby incorporated by.

The chemically reactive group of modified peptide can comprise with its different loci of reacting in vivo: cell, particularly blood rbc (red corpuscle) and thrombocyte; And protein, immunoglobulin (Ig) for example comprises that IgG and IgM, serum albumin, ferritin, steroid class are conjugated protein, Transferrins,iron complexes, thyroxine-binding protein, α-2-macroglobulin etc.Those and modified reactive polypeptide and the acceptor that can't survive were for a long time generally eliminated from the human body host in about 3 days.Based on the concentration in blood, above-mentioned protein (albumen that comprises cell) kept in blood flow 3 days at least, and can keep especially 5 days or the longer time (generally be no more than 60 days, more common be no more than 30 days) as the transformation period.

At first, reaction be with blood in flowable component carry out particularly blood protein and cell, more particularly blood protein and red corpuscle.So-called " flowing " be meant any long-time in fixing site not, generally be no more than 5 minutes, more normal is 1 minute, though some blood constitutents may be static relatively in for a long time.At first, should there be allogenic relatively functionalized albumen and cell colony.Yet in most cases, substantial variation will take place from initial colony in above-mentioned colony in a couple of days, and this depends on the transformation period of functionalized albumen in blood flow.So, normally about 3 days or the longer time in, IgG will become the main functionalized albumen in the blood flow.

Usually, after administration 5 days, IgG, serum albumin and red corpuscle in blood be the blood coupling component at least about 60mole%, usually at least about 75mole%, and IgG, IgM (substantially in low-down scope) and serum albumin be the acellular coupling component at least about 50mole%, usually at least about 75mole%, more usually be at least about 80mole%.

By use non-special modified peptide that modified peptide can make expection in vivo to the patient with and the conjugate of blood constitutent, this patient can be human body or other Mammalss.Administration can take bolus injection or by the long-time form of slowly introducing of infusion, utilize metered flow etc. to carry out.

If wish, also can make modified peptide and the reactive functionalities covalent bonding in the blood constitutent by blood and modified peptide of the present invention are mixed to come prepared ex vivo target conjugate, subsequently coupling blood is returned or is administered to the host.In addition, also the component by at first each blood constitutent of purifying or limited quantity such as red corpuscle, immunoglobulin (Ig), serum albumin wait and realize, and the modified peptide of these stripped components and chemical reactivity is combined in above-mentioned preparation.Subsequently functionalized blood or blood constitutent are returned to the host, provide goal treatment effective conjugate in vivo.In isolated operation, also can handle and prevent its aggegation blood.

5. Synthesizing of modified antiviral and antifusogenic peptides

A. Peptide is synthetic

The standard method of the solid-phase peptide chemistry that antiviral and/or antifusogenic peptides of the present invention can be known by one skilled in the art is synthetic.For example, peptide can by the solid state chemistry technology according to following by Steward and Young (Steward, J.M. and Young, J.D., " solid-phase peptide is synthetic " (Solid Phase Peptide Synthesis) the 2nd edition, Pierce Chemical Company, Rockford, III., (1984)) described method, utilize Applied Biosystem synthesizer preparation.Similarly, can synthesize a plurality of peptide fragment, be joined together to form bigger peptide fragment subsequently.These synthetic peptide fragments also carry out amino acid at specific position and replace.

Synthetic for solid-phase peptide, about the general introduction of multiple technologies can be referring to J.M.Stewart and J.D.Young, " solid-phase peptide is synthetic ", W.H.Freeman Co. (San Francisco), 1963 and J.Meienhofer, " neurophysin and peptide " (Hormonal Proteins and Peptide) vol 2, p.46, Academic Press (New York), 1973.Classical solution synthetic method can be referring to G.Schroder and K.Lupke, " peptide " (The Peptides), Vol.1, AcacemicPress (New York).Usually, these methods comprise that the amino acid whose order adding of one or more amino acid or due care makes peptide elongation.Generally, first amino acid whose amino or carboxyl are by suitable protecting group protection.Subsequently, the amino acid of this protection or derivatize is connected on the inertia solid phase carrier or the amino acid by adding the next due care that has complementation (amino or carboxyl) group in sequence and utilize being fit to form under the condition of amido linkage in solution.Remove protecting group and add next amino acid (due care) from initiate amino-acid residue subsequently, and so continue.

After the amino acid of whole expections has connected with suitable sequence, remove any residual protecting group (with any solid phase carrier) successively or simultaneously, obtain final polypeptide.By simple modifications to this universal method; add more than one amino acid on the chain in prolonging once; for example, by coupling (under the condition of the racemization chiral centre not) tripeptides of protection and the dipeptides of due care, behind deprotection, form pentapeptide.

The particularly preferred method for preparing The compounds of this invention comprises the solid phase method of peptide synthesis, wherein protects amino acid whose α-N-end with acid or alkali sensitive group.Such protecting group should have the characteristic of the conditional stability that peptide bond is formed, and is easy to simultaneously remove but can destroy the peptide chain of prolongation or can not make wherein contained any chiral centre racemization.The protecting group that is suitable for is: 9-fluorenyl methoxy carbonyl (Fmoc), tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), xenyl isopropoxy carbonyl, tert-pentyloxy carbonyl, isobornyl oxygen base carbonyl, α; alpha-alpha-dimethyl-3,5-dimethoxy benzyloxycarbonyl, ortho-nitrophenyl base sulfinyl, 2-cyano group-tert-butoxycarbonyl etc.9-fluorenyl-methoxycarbonyl (Fmoc) protecting group is particularly suitable for the synthetic of peptide of the present invention.Other preferred Side chain protective groups are: amino as Methionin and arginine for side chain, be 2,2,5,7,8-pentamethyl-benzo dihydropyrane-6-alkylsulfonyl (pmc), nitro, p-toluenesulfonyl, 4-anisole alkylsulfonyl, Cbz, Boc and adamantyl oxygen base carbonyl; For tyrosine, benzyl, adjacent bromo-benzyloxy-carbonyl, 2,6-dichloro benzyl, sec.-propyl, the tertiary butyl (t-Bu), cyclohexyl, cyclopentyl and ethanoyl (Ac); For Serine, the tertiary butyl, benzyl and THP trtrahydropyranyl; For Histidine, trityl, benzyl, Cbz, p-toluenesulfonyl and 2,4-dinitrophenyl; For tryptophane, formyl radical; For aspartic acid and L-glutamic acid, the benzyl and the tertiary butyl; With for halfcystine, trityl group (trityl).

In the solid phase method of peptide synthesis, α-C-end amino acid is connected on the suitable solid phase carrier or resin.The above-mentioned synthetic solid phase carrier that is suitable for is those to the reagent of gradual condensation-deprotection reaction and condition is inertia and undissolved material in used medium.The preferred solid phase carrier of synthetic institute of C-terminal carboxyl(group) peptide is 4-hydroxymethyl phenoxymethyl-copolymerization (vinylbenzene-1% Vinylstyrene).The preferred solid phase carrier of α-C-terminal amide peptide be Applied Biosystems (Foster City, Calif.) 4-of Chu Shouing (2 ', 4 '-Dimethoxyphenyl-Fmoc-amino methyl) phenoxy-acetamide base ethylamide resin.At dispensable 4-dimethylaminopyridine (DMAP), I-hydroxybenzotriazole (HOBT), benzotriazole-1-base oxygen base-three is (under dimethylamino) Phosphonium hexafluorophosphate (BOP) or two (2-oxo-3-oxazolidinyl) phosphonium chloride (BOPCl), α-C-end amino acid and resin are at N, N '-dicyclohexylcarbodiimide (DCC), N, N '-DIC (DIC) or O-benzotriazole-1-base-N, N, N ', N '-tetramethyl-urea-hexafluorophosphate (HBTU) mediation is down in 10 to 50 ℃ temperature, coupling about 1 was to about 24 hours in solvent (as methylene dichloride or DMF).

When solid phase carrier be 4-(2 ', 4 '-Dimethoxyphenyl-Fmoc-amino methyl) during phenoxy group-acetamido ethylamide resin, with above-mentioned α-C-end amino acid coupling before, the Fmoc base is with secondary amine, preferred piperidines cracking.With the 4-of deprotection (2 '; 4 '-Dimethoxyphenyl-Fmoc-amino methyl) phenoxy group-acetamido ethylamide resin link coupled preferred method is to utilize O-benzotriazole-1-base-N in DMF; N; N '; N '-tetramethyl-urea hexafluoro-phosphoric acid (HBTU; 1 equivalent) and I-hydroxybenzotriazole (HOBT, 1 equivalent).The amino acid whose coupling that success is protected can be carried out in the automatic Peptide synthesizer that affiliated field is known.In a preferred implementation, with the α--terminal amino acid of Fmoc protection growthing peptide chain.By handle the Fmoc protecting group of the α-N-end side that can slough growthing peptide chain with secondary amine, preferred piperidines.Subsequently the amino acid of various protections is introduced with about 3 times of molar excess, linked reaction suits to carry out in DMF.Coupling agent generally is O-benzotriazole-1-base-N, N, N ', N '-tetramethyl-urea phosphofluoric acid (HBTU, 1 equivalent) and I-hydroxybenzotriazole (HOBT, 1 equivalent).

When solid phase synthesis finished, polypeptide removed with deprotection or carries out continuously or carry out in an operation on the resin.Can utilize cracking agent process resin bonded polypeptide to finish removing of polypeptide and deprotection in same operation, cracking agent comprises thioanisole, water, ethane two mercaptan and trifluoroacetic acid.α-C-end at polypeptide is that resin is separated cracking with alkylamine by ammonia in the situation of alkane acid amides.In addition, peptide can pass through transesterification, for example uses methyl alcohol, separate by ammonia subsequently or directly amide group shift and remove.The peptide of protection is purifying at this moment, perhaps directly carries out next step.Removing of Side chain protective group is to utilize above-mentioned mixed pyrolysis agent to carry out.The peptide of deprotection carries out purifying by the chromatographic step of the following any or all of type of a series of employings fully: the ion-exchange on the weakly base resin (acetate type); Do not derive hydrophobic adsorption chromatography on the polystyrene-divinylbenzene (for example Amberlite XAD); Silica gel adsorption chromatography; Ion-exchange chromatography on the carboxymethyl cellulose; Distribution chromatography, for example on Sephadex G-25, LH-20 or adverse current distribute; High efficiency liquid chromatography (HPLC), especially octyl group-or octadecyl silyl-silica gel bonded phase column packed on reversed-phase HPLC.

The molecular weight of these ITP utilizes fast atom bombardment (FAB) mass spectrometric determination.

1.N-terminal protecting group

As mentioned above, term " N-protected base " is meant that the group of not expected response does not take place in building-up process those α that is used for protecting amino acid or peptide-N-amino terminal or protection amino acid or peptide.N-protected base commonly used is disclosed in " protecting group in the organic synthesis " (Protective Groupsin Organic Synthesis, John Wiley ﹠amp; Sons, New York (1981)), it is hereby incorporated by.In addition, protecting group can be used as prodrug and uses, and it is easy to discharge the biological activity parent by for example enzymically hydrolyse cracking in vivo.α-N-protected base comprises: lower alkane acyl group (loweralkanoyl), for example formyl radical, ethanoyl (" Ac "), propionyl, valeryl, tertiary butyl ethanoyl etc.; Other acyl groups, comprise 2-chloracetyl, 2-acetyl bromide, trifluoroacetyl group, tribromo-acetyl base, phthaloyl, ortho-nitrophenyl oxygen base ethanoyl ,-chlorobutyryl, benzoyl, 4-chlorobenzene formacyl, 4-benzoyl bromide, 4-nitro benzoyl etc.; Alkylsulfonyl, for example benzenesulfonyl, p-toluenesulfonyl etc.; Carbamate forms group, benzyloxycarbonyl for example, to the chlorine benzyloxycarbonyl, to methoxyl group benzyloxy base carbonyl, to the nitro benzyloxycarbonyl, 2-nitro benzyloxycarbonyl, to the bromo-benzyloxy-carbonyl, 3,4-dimethoxy benzyloxycarbonyl, 3,5-dimethoxy benzyloxycarbonyl, 2,4-dimethoxy benzyloxycarbonyl, 4-oxyethyl group benzyloxy base carbonyl, 2-nitro-4,5-dimethoxy benzyloxycarbonyl, 3,4,5-trimethoxy benzyloxycarbonyl, 1-(to xenyl)-1-methyl ethoxy carbonyl, α, alpha-alpha-dimethyl-3,5-dimethoxy benzyloxycarbonyl, diphenyl-methyl oxygen base carbonyl, tert-butoxycarbonyl (Boc), the di-isopropyl methoxycarbonyl, isopropoxy carbonyl, ethoxy carbonyl, methoxycarbonyl, allyloxy carbonyl, 2,2,2-trichlorine ethoxy carbonyl, phenyloxycarbonyl, the 4-nitrophenoxy carbonyl, fluorenyl-9-methoxycarbonyl, the cyclopentyloxy carbonyl, the Buddha's warrior attendant alkoxy carbonyl, cyclohexyl oxygen base carbonyl, phenyl thiocarbonyl etc.; Aralkyl, for example benzyl, trityl, benzyloxymethyl, 9-fluorenyl methoxy carbonyl (Fmoc) etc.; And silyl, for example trimethyl silyl etc.

2. carboxyl-protecting group

As mentioned above, term " carboxyl-protecting group " is meant carboxylic acid protection ester or the amide group that is used to intercept or protect carboxylic functionality when the reaction of other functional site that relate to compound.Carboxyl-protecting group is disclosed in Greene's " protecting group in the organic synthesis " pp.152-186 (1981), and it is hereby incorporated by.In addition, carboxyl-protecting group can be used as prodrug, and carboxyl-protecting group can be easy to cracking in vivo thus, for example by enzymically hydrolyse, discharges the biological activity parent.This type of carboxyl-protecting group is known for one of ordinary skill in the art, and they had been widely used in the carboxy protective in penicillin and the cynnematin field already, and as U.S. patent 3,840,556 and 3,719,667 is described, and its disclosure is hereby incorporated by.Typical carboxyl-protecting group is: C1-C8 low alkyl group (for example methyl, ethyl or the tertiary butyl etc.); Aralkyl, for example styroyl or benzyl and substitutive derivative thereof, for example alkoxybenzyl or nitrobenzyl etc.; Aromatic yl alkenyl, for example phenyl vinyl etc.; Aryl and substitutive derivative thereof, for example 5-dihydro indenyl etc.; Dialkyl aminoalkyl, for example dimethyl aminoethyl etc.); Alkanoyloxy alkyl, for example acetoxy-methyl, butyryl acyloxy methyl, valeryl oxygen ylmethyl, isobutyl acyl-oxygen ylmethyl, isoamyl acyl-oxygen ylmethyl, 1-(propionyloxy)-1-ethyl, 1-(new pentane acyloxy)-1-ethyl, 1-methyl isophthalic acid-(propionyloxy)-1-ethyl, pivalyl ethyl-methyl, propionyloxy methyl etc.; Cycloalkanes acyloxy alkyl, for example cyclopropyl carbonyl oxy-methyl, cyclobutyl carbonyl oxy-methyl, cyclopentyl carbonyl oxy-methyl, cyclohexyl carbonyl oxy-methyl etc.; Aryl acyloxy alkyl, for example benzoyloxy methyl, benzoyloxy ethyl etc.; Aralkyl carbonyl oxygen base alkyl, for example benzyl carbonyl oxy-methyl, 2-benzyl carbonyl oxygen base ethyl etc.; Alkoxy carbonyl alkyl or cyclo alkoxy carbonyl alkyl, for example methoxycarbonyl methyl, cyclohexyloxy carbonyl methyl, 1-methoxycarbonyl-1-ethyl etc.; Alkoxyl group carbonyl oxygen base alkyl or cycloalkyloxy carbonyl oxygen base alkyl, for example methoxyl group carbonyl oxy-methyl, tert.-butoxy carbonyl oxy-methyl, 1-oxyethyl group carbonyl Oxy-1-ethyl, 1-cyclohexyloxy carbonyl oxygen-1-ethyl etc.; Aryloxy carbonyl oxygen base alkyl, for example 2-(phenoxy group carbonyl oxygen base) ethyl, 2-(5-dihydro indenyl oxygen base carbonyl oxygen base) ethyl etc.; Alkoxyalkyl carbonyl oxygen base alkyl, for example 2-(1-methoxyl group-2-methyl-prop-2-acyloxy) ethyl etc.; Aralkoxy carbonyl oxygen base alkyl, for example 2-(benzyloxycarbonyloxy base) ethyl etc.; Aromatic yl alkenyl oxygen base carbonyl oxygen base alkyl, for example 2-(3-phenyl propylene-2-base oxygen base carbonyl oxygen base) ethyl etc.; Alkoxycarbonyl amino alkyl, for example tert-butoxycarbonyl amino methyl etc.; Alkyl amino-carbonyl aminoalkyl group, for example methylamino carbonylamino methyl etc.; Alkanoylamino alkyl, for example acetylamino methyl etc.; Heterocycle carbonyl oxygen base alkyl, for example 4-methylpiperazine base carbonyl oxy-methyl etc.; Dialkyl amino carbonyl alkyl, for example dimethylamino carbonyl methyl, diethylamino carbonyl methyl etc.; (5-(low alkyl group)-2-oxo-1,3-Dioxol-4-yl) alkyl, for example (the 5-tertiary butyl-2-oxo-1,3-Dioxol-4-yl) methyl etc.; (5-phenyl-2-oxo-1,3-Dioxol-4-yl) alkyl, for example (5-phenyl-2-oxo-1,3-Dioxol-4-yl) methyl etc.

Representational amido carboxyl protecting group is aminocarboxyl and low-grade alkyl amino carbonylic.

Preferred carboxy protective compound of the present invention is that the carboxyl of wherein protecting is low alkyl group, cycloalkyl or aralkyl ester, such as methyl esters, ethyl ester, propyl ester, isopropyl ester, butyl ester, secondary butyl ester, isobutyl ester, pentyl ester, isopentyl ester, monooctyl ester, cyclohexyl, phenethyl ester etc.; Or the compound of alkanoyloxy alkyl, cycloalkanes acyloxy alkyl, aryl acyloxy alkyl or aralkyl carbonyl oxygen base alkyl ester.Preferred amido carboxyl protecting group is a low-grade alkyl amino carbonylic.For example, aspartic acid can be terminal with acid-unstable group (for example tertiary butyl) protection and terminal with hydrogenation sensitive group (for example benzyl) protection at β-C-at α-C-, subsequently deprotection optionally in building-up process.

B. The modification of peptide

The present invention prepares the mode of modified peptide can carry out multiple variation in the essence of interior different factors according to containing peptide.The selection of synthetic method should be easy, high yield is provided and can accesses highly purified product.Usually, chemically reactive group should produce in the synthetic final stage, and for example, for carboxyl, esterification generates active ester.It is as described below that the present invention prepares the concrete grammar of modified peptide.

Particularly, at first measure the antiviral activity of selected peptide, only modify with linking group subsequently in the N of peptide end, C end or inside.Measure the antiviral activity of this modified peptide-linking group subsequently.If antiviral activity does not sharply reduce (promptly reducing less than 10 times), then utilize its volume lifetime to measure the stability of this modified peptide-linker.If stability is not improved and is come up to the expectation, then modify this peptide in another site, repeat this process until the antiviral level and the stability that obtain expection.

More particularly, selection stands each peptide of linker and reaction entity base group modification and should modify according to following standard: if the terminal carboxyl(group) on the peptide can utilize and the maintenance of enantiopathy cytotoxic activity does not have decisive role, there are not other responsive functional groups on the peptide simultaneously, the tie point of then selecting this carboxyl to modify as linker-reactive group.If terminal carboxyl(group) participates in antiviral activity, if or do not have carboxyl to utilize, then select any other tie point to keeping antiviral activity not have responsive functional group of materially affect to modify as linker-reaction entity.If there are several available responsive functional groups on peptide, the cooperative programs of protecting group should be used in a certain way, so that behind the deprotection of the adding of linker/reaction entity and all protected responsive functional groups, still can keep antiviral activity.If there is not responsive functional group to utilize on the peptide, perhaps wish to carry out simpler modification approach, should carry out in the mode that can keep antiviral activity for the synthetic practice of the modification of initial peptide.In this case, modify the opposite ends that occurs in peptide.

The NHS derivative can not exist under the condition of other responsive functional groups synthetic in peptide by carboxylic acid.Particularly, this peptide is at anhydrous CH ₂Cl ₂With among the EDC with N-hydroxy-succinamide reaction, product obtains the NHS derivative by chromatography or recrystallization from appropriate solvent system purifying.

In addition, the NHS derivative can be synthetic from the peptide that contains amino and/or sulfydryl and carboxylic acid.When having free amine group or sulfydryl in the molecule, preferably before adding the NHS derivative, these responsive protective groups are got up.For example, if molecule contains free amine group, be necessary to state on the implementation the amine that chemistry makes amine be converted into Fmoc before or preferably is converted into the tBoc protection.The amine functionality is not a deprotection after the preparation of NHS derivative.So this method only is applicable to that those needn't discharge its amino compound that produces the expection antiviral effect.Discharge the amino primary characteristic that keeps molecule if desired, another kind of chemistry below then having to carry out.

In addition, the NHS derivative can be from containing amino or sulfydryl but not have the peptide of carboxylic acid synthetic.When the molecule of selecting does not contain carboxylic acid, can utilize one group of difunctionality linker to make molecule be converted into reactive NHS derivative.For example, ethylene glycol-two (succinimido succinates) (EGS) are dissolved among the DMF with triethylamine,, produce single NHS derivative to wherein adding the molecule (being suitably 10: 1) that contains free amine group with the ratio of EGS.In order to prepare the NHS derivative, can utilize the N-[-dimaleoyl imino butyryl acyloxy that is present among the DMF from the sulfydryl derived molecules] succinimide ester (GMBS) and triethylamine.The reaction of dimaleoyl imino and free sulfhydryl groups utilizes on silica gel chromatography or can be from reaction mixture purifying NHS derivative by HPLC.

The NHS derivative also can be synthetic from the peptide that contains a plurality of responsive functional groups.Various situations are through analyzing and solving in a different manner.Yet; by means of a large amount of commercially available protecting group and difunctionality linkers; the present invention can be applied to any peptide; preferably modify described peptide (as mentioned above) by a chemical step, or two steps (comprising the protection formerly of sensitive group as mentioned above) or three steps (protection, activation and deprotection).Only in the situation of exception, need utilize the synthetic of multistep (surpassing three steps) that peptide is converted into active NHS or maleimide derivatives.

Maleimide derivatives also can be synthetic by the peptide that contains free amine group and free carboxy acid.In order to prepare maleimide derivatives, can utilize the N-[γ-dimaleoyl imino butyryl acyloxy that is present among the DMF from the amino derivatization molecule] succinimide ester (GMBS) and triethylamine.The reaction of succinimide ester group and free amine group, and by crystallization or on silica gel chromatography or HPLC can be from reaction mixture the purifying maleimide derivatives.

At last, maleimide derivatives can be synthetic from containing a plurality of other responsive functional groups and not having free carboxy acid's peptide.The branch period of the day from 11 p.m. to 1 a.m when selecting not contain carboxylic acid can utilize one group of bifunctional cross-linker that this molecule is converted into reactive NHS derivative.For example, the reaction of the carboxyl by unhindered amina and MPA, utilize the HBTU/HOBt/DIEA activation, dimaleoyl imino propionic acid (MPA) can generate maleimide derivatives with the unhindered amina coupling in DMF.

Many other commercially available Heterobifunctional linking agents can be where necessary as an alternative.A large amount of difunctional compounds can be used for connecting entity.The example of reagent comprises: the triazobenzene formyl hydrazine; N-[4-(to azido-salicyl amino) butyl]-3 '-[2 '-the pyridyl dithio) propionic acid amide); two sulfosuccinimide base suberates; dimethyl adipimide ester; two succinimido tartrates; N-y-dimaleoyl imino butyryl acyloxy succinimide ester; N-hydroxysulphosuccinimide base-4-triazobenzene manthanoate; N-succinimido [4-azido-phenyl]-1,3 '-the dithio propionic ester; N-succinimido [4-iodo ethanoyl] Aminobenzoate; glutaraldehyde and succinimido 4-[N-maleimide ylmethyl] hexanaphthene-1-manthanoate.

6. The application of modified antiviral peptide

Modified antiviral peptide of the present invention can be used as therapeutical agent in the patient's who suffers from virus infection treatment, and can be according to method described below and other currently known methods of affiliated field to patient's administration.The genotoxic potential of peptide can be measured and should consider to the dose therapeutically effective of modified peptide by well-known to one skilled in the art method.

Modified peptide also can preventatively give formerly not infected individuals.This is of value to the dangerous high individual instances of those contact viruses, and this may appear at those and carry in the contacted individuality of infected individuals of high-risk viral communication.This is particularly advantageous in the treatment of those known viruses (as HIV virus).For example, the preventive administration of modified anti-HIV peptide will be of value to those situations that has contacted the healthcare worker of HIV infected individuals blood, or be engaged in the high-risk active individual instances that may contact HIV virus at other.

7. The administration of modified antiviral and antifusogenic peptides

Usually, modified peptide should be present in when administration in the physiology acceptable medium, for example deionized water, phosphate buffered saline (PBS) (PBS), salt solution, aqueous ethanol or other alcohol, blood plasma, albumen solution, mannitol, G/W, alcohol, plant wet goods.Other additive that can contain comprises: buffer reagent, its medium generally are buffered in about 5 to 10 the pH scope, and the concentration of buffer reagent generally is about 50 to 250mM; Salt, wherein the concentration of salt is about 5 usually to 500mM; Physiology can be accepted stablizer etc.Composition can be frozen drying so that conventional the storage and transportation.

Modified peptide majority is (IV), intra-arterial (IA), intramuscular (IM), subcutaneous (SC) etc. in parenterai administration such as blood vessel.Suitably can pass through the transfusion administration in the situation.In some occasion, when the reacting phase of functional group when slow, can be by oral, intranasal, rectum, transdermal or aerosol administration, the characteristic of conjugate allows to be delivered to vascular system thus.Usually should adopt single dose injection, though can adopt more than the injection once if desired the time.Modified peptide can utilize any instrument administration, comprises syringe, trochar, intubate etc.

Concrete administering mode should be according to dosage, single dose bolus injection or successive administration etc. change.More preferably, answer intravascular administration, the site of introducing is unimportant for the present invention, and preferred site is blood flow position rapidly, for example intravenously, periphery or maincenter vein.Find also can utilize other approach, wherein administration combines with slow release method or protectiveness matrix phase.The intention of doing like this is, modified peptide effectively is distributed in the blood, can react with blood constitutent thus.The concentration of conjugate is about 1pg/ml to 50mg/ml usually with changeful.The total amount of intravascular administration generally should be at about 0.1mg/ml to about 10mg/ml, Chang Weiyue 1mg/ml extremely in the scope of about 5mg/ml more.

By combining, a plurality of superiority have been produced with long lifetime blood constitutent such as immunoglobulin (Ig), serum albumin, red corpuscle and thrombocyte.The activity of described peptide is lengthened to a couple of days to several weeks.During this period of time only need be administered once.Can obtain higher specificity,, as if seldom be absorbed into other physiological process of cell internal interference thus because active compound is main and the macromole bonding.

8. Monitor the existence of modified peptide

Can monitor the blood one or many of mammalian hosts for the existence of monitoring modified peptide.Change into and have therapeutic activity by gathering the sample of a part or host's blood, can determine peptide that whether q.s is arranged and long lifetime blood constitutent bonding, and can measure the level of peptide compounds in the blood subsequently.If wish, can also measure peptide actually with which kind of blood constitutent bonding.This is extremely important when adopting non-specific modified peptide.For the peptide of specificity maleimide-modification, the transformation period of calculating serum albumin and IgG can be simpler.

A. Immunoassay

Another aspect of the present invention relates to the method for measuring antiviral peptide and/or analogue or derivatives thereof and the concentration of conjugate in biological specimen (for example blood), and this method has been utilized described peptide, peptide analogs or derivatives thereof and the specific antibody of conjugate; And relate to the application of this antibody-like in the toxic treatment relevant with described peptide, analogue and/or its derivative or conjugate possibility.This is useful, because body internal stability and life-span that described peptide has improved may cause new problem in treatment, comprises the increase of poisoning possibility.

The application that particular peptide, peptide analogs or derivatives thereof is had specific mono-clonal or polyclonal treatment-resistant agent antibody can help to solve this type of problem.Antibody can produce or derived from the host with particular peptide, the immunity of analogue or derivatives thereof, or with the immunogenic fragments of reagent or with the host of the corresponding synthetic immunogen immunity of the immunologic determinants of reagent.Preferred antibody reply natural, peptide modified and the coupling form has the specificity and the avidity of height.This antibody-like also can carry out mark with enzyme, fluorescence dye or radioactively labelled substance.

Can prepare with the peptide of purifying modified peptide is had specific antibody, thereby induce peptide specific antibody.By inducing of antibody, not only through to can immune response stimulating after the animal injection, and in the preparation of synthetic antibody or other specific binding molecules also to carry out similar step, for example screen the recombination immunoglobulin library.Mono-clonal and polyclonal antibody can prepare by the method that affiliated field is known.

Can also utilize anti-peptide antibody to treat the poisoning that causes by modified peptide, its analogue or derivative administration, and can exsomatize or the interior use of body.Stripped method comprises utilizes the treatment-resistant agent antibody that is fixed on the solid phase carrier that immune dialysis treatment is carried out in poisoning.The interior method of body comprises with the amount of the removing of effective initiation antibody-therapeutical agent mixture uses treatment-resistant agent antibody.

Contact with antibody by the blood of ordering at sterilising conditions, can remove patient exsomatize modified peptide or its analogue or conjugate in the blood with antibody.For example, antibody can be fixed on the base for post matter, and blood samples of patients is taken out and this matrix of process from the patient.Modified peptide, peptide analogs, derivative or conjugate will with this antibodies, the blood that contains peptide, analogue, derivative or the conjugate of lower concentration can turn back in patient's recycle system.Can control the amount of removing of peptide compounds by regulating pressure and flow.

Described peptide, analogue, derivative and conjugate are preferentially removed from the plasma component of blood samples of patients and can be affected, and for example, utilize semi-permeable membranes; Or the currently known methods in field separates cellular component and plasma component under at first utilizing, and makes plasma component flow through containing the matrix of treatment-resistant agent antibody subsequently.In addition, peptide link coupled hemocyte (comprising red corpuscle) preferentially remove can by collect with concentrated blood samples of patients in hemocyte and the serum component that the anti-ITP antibody of these cells and fixed is contacted with the eliminating blood samples of patients be affected.

Treatment-resistant agent antibody can be administered to the patient who accepts described peptide, analogue, derivative or conjugate treatment in non-enteron aisle body.Antibody will combine with peptide compounds and conjugate.In case combination, even be not to block fully, the activity of peptide also will be hindered, reduce thus peptide compounds in patient's blood flow biological effective concentration and alleviate deleterious side effect.In addition, bonded antibody-peptide complex will help the removing of peptide compounds and conjugate in patient's blood flow.

The present invention who describes now illustrates by following non-limiting examples comprehensively.

Embodiment 1

The preparation of modified DP178--

YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNWFK (MPA)-NH ₂Synthetic

In the present embodiment, synthetic and modify DP178 (SEQ IDNO:1) according to following synthetic route for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, DP178 is effective inhibitor of HIV-1, and can suppress that HIV-1 infects and non-infected cells between the synplasm of cell induction form and non-infected cells by the infection of acellular HIV-1 virus.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer (Symphony Peptide Synthesizer) of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH; Fmoc-Phe-OH; Fmoc-Trp (Boc)-OH; Fmoc-Asn (Trt)-OH; Fmoc-Trp (Boc)-OH; Fmoc-Leu-OH; Fmoc-Ser (tBu)-OH; Fmoc-Ala-OH; Fmoc-Trp (Boc)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Asp (tBu)-OH; Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH; Fmoc-Gln (Trt)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Met-OH; Fmoc-Lys (Boc)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Asn (Trt)-OH; Fmoc-Lys (Boc)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Asn (Trt)-OH; Fmoc-Gln (Trt)-OH, Fmoc-Ser (tBu)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Ser (tBu)-OH; Fmoc-His (Boc)-OH; Fmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Ser (tBu)-OH; Fmoc-Thr (tBu)-OH, Fmoc-Tyr (tBu)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.During end of synthesis, by being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; The modified peptide (being DAC) that uses the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm to obtain expecting is measured purity＞95% of peptide by RP-HPLC.

Embodiment 2

The preparation of modified DP107--

NNLLRAIEAQQHLLQLTVWQIKQLQARILAVERYLKDQK (MPA) NH ₂Synthetic

In the present embodiment, synthetic and modify DP107 (SEQ IDNO:2) according to following synthetic route for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and DP107 shows the effective antiviral activity to HIV-1.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH; Fmoc-Gln (Trt)-OH; Fmoc-Asp (tBu)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Leu-OH; Fmoc-Tyr (tBu)-OH; Fmoc-Arg (Pbf)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Val-OH; Fmoc-Ala-OH; Fmoc-Leu-OH, Fmoc-Ile-OH, Fmoc-Arg (Pbf)-OH; Fmoc-Ala-OH; Fmoc-Gln (Trt)-OH, Fmoc-Leu-OH, Fmoc-Gln (Trt)-OH; Fmoc-Lys (Boc)-OH; Fmoc-Ile-OH, Fmoc-Gln (Trt)-OH, Fmoc-Trp (Boc)-OH; Fmoc-Val-OH; Fmoc-Thr (tBu)-OH, Fmoc-Leu-OH, Fmoc-Gln (Trt)-OH; Fmoc-Leu-OH; Fmoc-Leu-OH, Fmoc-His (Boc)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Gln (Trt)-OH; Fmoc-Ala-OH, Fmoc-Glu (tBu)-OH, Fmoc-Ile-OH; Fmoc-Ala-OH; Fmoc-Arg (Pbf)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH; Fmoc-Asn (Trt)-OH, Fmoc-Asn (Trt)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; The modified peptide (being DAC) that uses the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm to obtain expecting is measured purity＞95% of peptide by RP-HPLC.

Embodiment 3

The preparation of modified anti-RSV peptide (C-terminal 1)

In the present embodiment, according to following synthetic route with peptide

VITIELSNIKENKCNGAKVKLIKQELDKYKNAV (SEQ ID NO:16) is modified to and contains linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and native sequences (SEQ ID NO) suppresses the viral infection of respiratory syncytial virus (RSV), comprises that the fusion and the synplasm that suppress rsv infection and do not infect between the Hep-2 cell form.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH, Fmoc-Val-OH, Fmoc-Ala-OH; Fmoc-Asn (Trt)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Tyr (tBu)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Asp (tBu)-OH, Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Lys (Boc)-OH; Fmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH; Fmoc-Val-OH, Fmoc-Lys (Boc)-OH, Fmoc-Ala-OH; Fmoc-Gly-OH; Fmoc-Asn (Trt)-OH, Fmoc-Cys (Trt)-OH, Fmoc-Lys (Boc)-OH; Fmoc-Asn (Trt)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Ile-OH; Fmoc-Asn (Trt)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH; Fmoc-Ile-OH; Fmoc-Thr (tBu)-OH, Fmoc-Ile-OH, Fmoc-Val-OH..They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; The modified peptide (being DAC) that uses the UV detector (VarianDynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm to obtain expecting is measured purity＞95% of peptide by RP-HPLC.

Embodiment 4

The preparation of modified anti-RSV peptide (T-N end)

In the present embodiment, according to the synthetic peptide of following synthetic route

VITIELSNIKENKCNGAKVKLIKQELDKYKNAV (SEQ ID NO:17), its corresponding to the peptide of SEQ ID NO:16 but halfcystine (C) replaced by methionine(Met) (M) residue, and be modified to and contain linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and native sequences (SEQ ID NO:16) suppresses the viral infection of respiratory syncytial virus (RSV), comprises that the fusion and the synplasm that suppress rsv infection and do not infect between the Hep-2 cell form.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Val-OH, Fmoc-Ala-OH, Fmoc-Asn (Trt)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Tyr (tBu)-OH, Fmoc-Lys (Boc)-OH; Fmoc-Asp (tBu)-OH, Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH; Fmoc-Gln (Trt)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Ile-OH; Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH, Fmoc-Val-OH; Fmoc-Lys (Boc)-OH; Fmoc-Ala-OH, Fmoc-Gly-OH, Fmoc-Asn (Trt)-OH; Fmoc-Met-OH; Fmoc-Lys (Boc)-OH, Fmoc-Asn (Trt)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Lys (Boc)-OH; Fmoc-Ile-OH, Fmoc-Asn (Trt)-OH, Fmoc-Ser (tBu)-OH; Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH, Fmoc-Ile-OH, Fmoc-Thr (tBu)-OH; Fmoc-Ile-OH, Fmoc-Val-OH..They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; The modified peptide (being DAC) that uses the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm to obtain expecting is measured purity＞95% of peptide by RP-HPLC.

Embodiment 5

The preparation of modified anti-RSV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:14 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:14 suppresses the viral infection of respiratory syncytial virus (RSV), comprises that the fusion and the synplasm that suppress rsv infection and do not infect between the Hep-2 cell form.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Tyr (tBu)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Asp (tBu)-OH, Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Lys (Boc)-OH; Fmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH; Fmoc-Val-OH, Fmoc-Lys (Boc)-OH, Fmoc-Ala-OH; Fmoc-Gly-OH, Fmoc-Asn (Trt)-OH, Fmoc-Cys (Trt)-OH; Fmoc-Lys (Boc)-OH; Fmoc-Asn (Trt)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Lys (Boc)-OH; Fmoc-Ile-OH; Fmoc-Asn (Trt)-OH, Fmoc-Ser (tBu)-OH, Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH; Fmoc-Ile-OH, Fmoc-Thr (tBu)-OH, Fmoc-Ile-OH; Fmoc-Val-OH; Fmoc-Ser (tBu)-OH, Fmoc-Thr (tBu)-OH, Fmoc-Tyr (tBu)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (VarianDynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 6 (T-143)

The preparation of modified anti-RSV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:15 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:15 suppresses the viral infection of respiratory syncytial virus (RSV), comprises that the fusion and the synplasm that suppress rsv infection and do not infect between the Hep-2 cell form.

The solid-phase peptide of the modified peptide analogs on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH, Fmoc-Asn (Trt)-OH, Fmoc-Lys (Boc)-OH; Fmoc-Tyr (tBu)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Asp (tBu)-OH; Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Ile-OH, Fmoc-Leu-OH; Fmoc-Lys (Boc)-OH, Fmoc-Val-OH, Fmoc-Lys (Boc)-OH; Fmoc-Ala-OH, Fmoc-Gly-OH, Fmoc-Asn (Trt)-OH; Fmoc-Cys (Trt)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Asn (Trt)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Lys (Boc)-OH; Fmoc-Ile-OH, Fmoc-Asn (Trt)-OH, Fmoc-Ser (tBu)-OH; Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH, Fmoc-Ile-OH, Fmoc-Thr (tBu)-OH; Fmoc-Ile-OH; Fmoc-Val-OH, Fmoc-Ser (tBu)-OH, Fmoc-Thr (tBu)-OH..They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mLCHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 7

The preparation of modified anti-RSV peptide(C-terminal)

In the present embodiment, according to the synthetic peptide SEQ ID NO:17 of following synthetic route), its corresponding to its corresponding to the peptide of SEQ ID NO:16 but halfcystine (C) replaced by methionine(Met) (M) residue, and be modified to and contain linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and native sequences SEQ ID NO:16 suppresses the viral infection of respiratory syncytial virus (RSV), comprises that the fusion and the synplasm that suppress rsv infection and do not infect between the Hep-2 cell form.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH, Fmoc-Val-OH, Fmoc-Ala-OH; Fmoc-Asn (Trt)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Tyr (tBu)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Asp (tBu)-OH, Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Lys (Boc)-OH; Fmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH; Fmoc-Val-OH, Fmoc-Lys (Boc)-OH, Fmoc-Ala-OH; Fmoc-Gly-OH; Fmoc-Asn (Trt)-OH, Fmoc-Met-OH, Fmoc-Lys (Boc)-OH; Fmoc-Asn (Trt)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Ile-OH; Fmoc-Asn (Trt)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH; Fmoc-Ile-OH; Fmoc-Thr (tBu)-OH, Fmoc-Ile-OH, Fmoc-Val-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; The modified peptide (being DAC) that uses the UV detector (VarianDynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm to obtain expecting is measured purity＞95% of peptide by RP-HPLC.

Embodiment 8

The preparation of modified anti-RSV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:29 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:29 suppresses the viral infection of respiratory syncytial virus (RSV), comprises that the fusion and the synplasm that suppress rsv infection and do not infect between the Hep-2 cell form.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Asp (tBu)-OH; Fmoc-Ile-OH, Fmoc-Tyr (tBu)-OH, Fmoc-Asn (Trt)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Leu-OH, Fmoc-Asp (tBu)-OH; Fmoc-Leu-OH, Fmoc-Val-OH, Fmoc-Lys (Boc)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Thr (tBu)-OH, Fmoc-Leu-OH; Fmoc-Val-OH, Fmoc-Ser (tBu)-OH, Fmoc-Val-OH; Fmoc-Gly-OH; Fmoc-Asn (Trt)-OH, Fmoc-Ser (tBu)-OH, Fmoc-Leu-OH; Fmoc-Ser (tBu)-OH; Fmoc-Val-OH, Fmoc-Val-OH, Fmoc-Ala-OH; Fmoc-Lys (Boc)-OH; Fmoc-Asn (Trt)-OH, Fmoc-Thr (tBu)-OH, Fmoc-Ser (tBu)-OH; Fmoc-Leu-OH; Fmoc-Leu-OH, Fmoc-Ala-OH, Fmoc-Ile-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (VarianDynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 9 (T-173)

The preparation of modified anti-HPIV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:52 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:52 suppresses human parainfluenza virus's 3 (HPIV3) viral infection, comprises that the fusion and the synplasm that suppress HPIV3 infection Hep-2 cell and do not infect between the CV-1W cell form.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH, Fmoc-Ile-OH, Fmoc-Leu-OH; Fmoc-Asn (Trt)-OH, Fmoc-Gly-OH, Fmoc-Ile-OH; Fmoc-Ser (tBu)-OH, Fmoc-Ser (tBu)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Val-OH, Fmoc-Ser (tBu)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Val-OH, Fmoc-Ala-OH, Fmoc-Lys (Boc)-OH; Fmoc-Asn (Trt)-OH, Fmoc-Thr (tBu)-OH, Fmoc-Asp (tBu)-OH; Fmoc-Arg (Pbf)-OH, Fmoc-Ile-OH, Fmoc-Ala-OH; Fmoc-Glu (tBu)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Leu-OH; Fmoc-Lys (Boc)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Ile-OH, Fmoc-Asp (tBu)-OH; Fmoc-Ser (tBu)-OH; Fmoc-Arg (Pbf)-OH, Fmoc-Ala-OH, Fmoc-Gln (Trt)-OH; Fmoc-Lys (Boc)-OH; Fmoc-Ala-OH, Fmoc-Glu (tBu)-OH, Fmoc-Val-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 10

The preparation of modified anti-HPIV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:58 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:58 suppresses human parainfluenza virus's 3 (HPIV3) viral infection, comprises that the fusion and the synplasm that suppress HPIV3 infection Hep-2 cell and do not infect between the CV-1W cell form.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin:

Fmoc-Lys(Aloc)-OH，Fmoc-Val-OH，Fmoc-Ser(tBu)-OH，Fmoc-Lys(Boc)-OH，Fmoc-Ile-OH，Fmoc-Ala-OH，Fmoc-Val-OH，Fmoc-Ile-OH，Fmoc-Leu-OH，Fmoc-Asn(Trt)-OH，Fmoc-Gly-OH，Fmoc-Ile-OH，Fmoc-Ser(tBu)-OH，Fmoc-Ser(tBu)-OH，Fmoc-Gln(Trt)-OH，Fmoc-Val-OH，Fmoc-Ser(tBu)-OH，Fmoc-Gln(Trt)-OH，Fmoc-Val-OH，Fmoc-Ala-OH，Fmoc-Lys(Boc)-OH，Fmoc-Asn(Trt)-OH，Fmoc-Thr(tBu)-OH，Fmoc-Asp(tBu)-OH，Fmoc-Arg(Pbf)-OH，Fmoc-Ile-OH，Fmoc-Ala-OH，Fmoc-Glu(tBu)-OH，Fmoc-Lys(Boc)-OH，Fmoc-Leu-OH，Fmoc-Lys(Boc)-OH，Fmoc-Glu(tBu)-OH，Fmoc-Ile-OH，Fmoc-Asp(tBu)-OH，Fmoc-Ser(tBu)-OH，Fmoc-Arg(Pbf)-OH。They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 11

The preparation of modified anti-HPIV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:35 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:35 suppresses human parainfluenza virus's 3 (HPIV3) viral infection, comprises that the fusion and the synplasm that suppress HPIV3 infection Hep-2 cell and do not infect between the CV-1W cell form.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH; Fmoc-Leu-OH; Fmoc-Lys (Boc)-OH; Fmoc-Gln (Trt)-OH; Fmoc-Asn (Trt)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Arg (Pbf)-OH, Fmoc-Arg (Pbf)-OH; Fmoc-Ile-OH; Fmoc-Trp (Boc)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Lys (Boc)-OH; Fmoc-Ser (tBu)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH; Fmoc-Asp (tBu)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Ala-OH; Fmoc-Lys (Boc)-OH; Fmoc-Asn (Trt)-OH, Fmoc-Leu-OH, Fmoc-Ile-OH; Fmoc-Glu (tBu)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Ile-OH, Fmoc-Asp (tBu)-OH; Fmoc-Ile-OH; Fmoc-Pro-OH, Fmoc-Asp (tBu)-OH, Fmoc-Leu-OH; Fmoc-Gln (Trt)-OH Fmoc-Ala-OH; Fmoc-Val-OH, Fmoc-Ser (tBu)-OH, Fmoc-Asn (Trt)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 12

The preparation of modified anti-HPIV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:38 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:38 suppresses human parainfluenza virus's 3 (HPIV3) viral infection, comprises that the fusion and the synplasm that suppress HPIV3 infection Hep-2 cell and do not infect between the CV-1W cell form.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Ile-OH; Fmoc-Ser (tBu)-OH; Fmoc-Asp (tBu)-OH; Fmoc-Leu-OH; Fmoc-Lys (Boc)-OH; Fmoc-Gln (Trt)-OH; Fmoc-Asn (Trt)-OH, Fmoc-Ser (tBu)-OH, Fmoc-Arg (Pbf)-OH; Fmoc-Arg (Pbf)-OH; Fmoc-Ile-OH, Fmoc-Trp (Boc)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Lys (Boc)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH; Fmoc-Asp (tBu)-OH, Fmoc-Ser (tBu)-OH, Fmoc-Lys (Boc)-OH; Fmoc-Ala-OH; Fmoc-Lys (Boc)-OH, Fmoc-Asn (Trt)-OH, Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH; Fmoc-Ile-OH, Fmoc-Ser (tBu)-OH, Fmoc-Ile-OH; Fmoc-Asp (tBu)-OH; Fmoc-Ile-OH, Fmoc-Pro-OH, Fmoc-Asp (tBu)-OH; Fmoc-Leu-OH, Fmoc-Ala-OHBOC-Lys (Aloc)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (VarianDynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 13

The preparation of modified anti-HPIV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:39 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:39 suppresses human parainfluenza virus's 3 (HPIV3) viral infection, comprises that the fusion and the synplasm that suppress HPIV3 infection Hep-2 cell and do not infect between the CV-1W cell form.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH; Fmoc-Gly-OH; Fmoc-Ile-OH; Fmoc-Ser (tBu)-OH; Fmoc-Asp (tBu)-OH; Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH, Fmoc-Gly-OH; Fmoc-Asn (Trt)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Arg (Pbf)-OH, Fmoc-Arg (Pbf)-OH; Fmoc-Ile-OH; Fmoc-Trp (Boc)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Lys (B oc)-OH; Fmoc-Ser (tBu)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH; Fmoc-Asp (tBu)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Ala-OH; Fmoc-Lys (Boc)-OH; Fmoc-Asn (Trt)-OH, Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH; Fmoc-Ile-OH; Fmoc-Ser (tBu)-OH, Fmoc-Ile-OH, Fmoc-Asp (tBu)-OH; Fmoc-Ile-OH; Fmoc-Pro-OH, Fmoc-Asp (tBu)-OH, Fmoc-Leu-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (VarianDynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 14

The preparation of modified anti-HPIV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:40 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:40 suppresses human parainfluenza virus's 3 (HPIV3) viral infection, comprises that the fusion and the synplasm that suppress HPIV3 infection Hep-2 cell and do not infect between the CV-1W cell form.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin:

Fmoc-Lys(Aloc)-OH，Fmoc-Asn(Trt)-OH，Fmoc-Gly-OH，Fmoc-Ile-OH，Fmoc-Ser(tBu)-OH，Fmoc-Asp(tBu)-OH，Fmoc-Leu-OH，Fmoc-Lys(Boc)-OH，Fmoc-Gly-OH，Fmoc-Asn(Trt)-OH，Fmoc-Ser(tBu)-OH，Fmoc-Arg(Pbf)-OH，Fmoc-Arg(Pbf)-OH，Fmoc-Ile-OH，Fmoc-Trp(Boc)-OH，Fmoc-Glu(tBu)-OH，Fmoc-Lys(Boc)-OH，Fmoc-Ser(tBu)-OH，Fmoc-Glu(tBu)-OH，Fmoc-Glu(tBu)-OH，Fmoc-Leu-OH，Fmoc-Asp(tBu)-OH，Fmoc-Ser(tBu)-OH，Fmoc-Lys(Boc)-OH，Fmoc-Ala-OH，Fmoc-Lys(Boc)-OH，Fmoc-Asn(Trt)-OH，Fmoc-Leu-OH，Fmoc-Glu(tBu)-OH，Fmoc-Ile-OH，Fmoc-Ser(tBu)-OH，Fmoc-Ile-OH，Fmoc-Asp(tBu)-OH，Fmoc-Ile-OH，Fmoc-Pro-OH，Fmoc-Asp(tBu)-OH。They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mLCHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 15

The preparation of modified anti-HPIV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:41 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:41 suppresses human parainfluenza virus's 3 (HPIV3) viral infection, comprises that the fusion and the synplasm that suppress HPIV3 infection Hep-2 cell and do not infect between the CV-1W cell form.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Trp (Boc)-OH; Fmoc-Asn (Trt)-OH; Fmoc-Gly-OH; Fmoc-Ile-OH; Fmoc-Ser (tBu)-OH; Fmoc-Asp (tBu)-OH; Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH, Fmoc-Gly-OH; Fmoc-Asn (Trt)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Arg (Pbf)-OH, Fmoc-Arg (Pbf)-OH; Fmoc-Ile-OH; Fmoc-Trp (Boc)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Lys (Boc)-OH; Fmoc-Ser (tBu)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH; Fmoc-Asp (tBu)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Ala-OH; Fmoc-Lys (Boc)-OH; Fmoc-Asn (Trt)-OH, Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH; Fmoc-Ile-OH; Fmoc-Ser (tBu)-OH, Fmoc-Ile-OH, Fmoc-Asp (tBu)-OH; Fmoc-Ile-OH, Fmoc-Pro-OH Boc-Lys (Aloc)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mLCHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 16

The preparation of modified anti-HPIV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:42 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:42 suppresses human parainfluenza virus's 3 (HPIV3) viral infection, comprises that the fusion and the synplasm that suppress HPIV3 infection Hep-2 cell and do not infect between the CV-1W cell form.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH; Fmoc-His (Boc)-OH; Fmoc-Trp (Boc)-OH; Fmoc-Asn (Trt)-OH; Fmoc-Gly-OH; Fmoc-Ile-OH, Fmoc-Ser (tBu)-OH, Fmoc-Asp (tBu)-OH; Fmoc-Leu-OH; Fmoc-Lys (Boc)-OH, Fmoc-Gly-OH, Fmoc-Asn (Trt)-OH; Fmoc-Ser (tBu)-OH; Fmoc-Arg (Pbf)-OH, Fmoc-Arg (Pbf)-OH, Fmoc-Ile-OH; Fmoc-Trp (Boc)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Ser (tBu)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH, Fmoc-Asp (tBu)-OH; Fmoc-Ser (tBu)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Ala-OH, Fmoc-Lys (Boc)-OH; Fmoc-Asn (Trt)-OH; Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH, Fmoc-Ile-OH; Fmoc-Ser (tBu)-OH; Fmoc-Ile-OH, Fmoc-Asp (tBu)-OH, Fmoc-Ile-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 17

The preparation of modified anti-MeV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:77 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:77 suppresses the viral infection of Measles virus (MeV), comprises suppressing MeV infection and not fusion between the vero cells infection and synplasm formation.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH; Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH, Fmoc-Lys (Boc)-OH; Fmoc-Ala-OH, Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH; Fmoc-Lys (Boc)-OH, Fmoc-Ala-OH, Fmoc-Ile-OH; Fmoc-Ala-OH, Fmoc-Asn (Trt)-OH, Fmoc-Gly-OH; Fmoc-Leu-OH, Fmoc-Asn (Trt)-OH, Fmoc-Thr (tBu)-OH; Fmoc-Gly-OH, Fmoc-Val-OH, Fmoc-Asp (tBu)-OH; Fmoc-Leu-OH; Fmoc-Arg (Pbf)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH; Fmoc-Ser (tBu)-OH; Fmoc-Ile-OH, Fmoc-Pro-OH, Fmoc-Pro-OH; Fmoc-Gly-OH; Fmoc-Leu-OH, Fmoc-Asp (tBu)-OH, Fmoc-Ile-OH; Fmoc-Arg (Pbf)-OH, Fmoc-His (Boc)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 18

The preparation of modified anti-MeV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:79 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:79 suppresses the viral infection of Measles virus (MeV), comprises suppressing MeV infection and not fusion between the vero cells infection and synplasm formation.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH, Fmoc-Ser (tBu)-OH, Fmoc-Ser (tBu)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Ala-OH; Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH; Fmoc-Ala-OH, Fmoc-Ile-OH, Fmoc-Ala-OH; Fmoc-Asn (Trt)-OH, Fmoc-Gly-OH, Fmoc-Leu-OH; Fmoc-Asn (Trt)-OH, Fmoc-Thr (tBu)-OH, Fmoc-Gly-OH; Fmoc-Val-OH; Fmoc-Asp (tBu)-OH, Fmoc-Leu-OH, Fmoc-Arg (Pbf)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH, Fmoc-Ser (tBu)-OH, Fmoc-Ile-OH; Fmoc-Pro-OH; Fmoc-Pro-OH, Fmoc-Gly-OH, Fmoc-Leu-OH; Fmoc-Asp (tBu)-OH, Fmoc-Ile-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 19

The preparation of modified anti-MeV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:81 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:79 suppresses the viral infection of Measles virus (MeV), comprises suppressing MeV infection and not fusion between the vero cells infection and synplasm formation.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Asp (tBu)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Ser (tBu)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Ala-OH, Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH, Fmoc-Ala-OH; Fmoc-Ile-OH, Fmoc-Ala-OH, Fmoc-Asn (Trt)-OH; Fmoc-Gly-OH, Fmoc-Leu-OH, Fmoc-Asn (Trt)-OH; Fmoc-Thr (tBu)-OH; Fmoc-Gly-OH, Fmoc-Val-OH, Fmoc-Asp (tBu)-OH; Fmoc-Leu-OH; Fmoc-Arg (Pbf)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH; Fmoc-Ser (tBu)-OH; Fmoc-Ile-OH, Fmoc-Pro-OH, Fmoc-Pro-OH; Fmoc-Gly-OH, Fmoc-Leu-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 20

The preparation of modified anti-MeV peptide

In the present embodiment, according to following synthetic route synthetic and modified peptides SEQ ID NO:84 for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:84 suppresses the viral infection of Measles virus (MeV), comprises suppressing MeV infection and not fusion between the vero cells infection and synplasm formation.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH, Fmoc-Arg (Pbf)-OH, Fmoc-Leu-OH; Fmoc-Ile-OH, Fmoc-Gln (Trt)-OH, Fmoc-Asp (tBu)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Ser (tBu)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Ala-OH, Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH, Fmoc-Ala-OH; Fmoc-Ile-OH, Fmoc-Ala-OH, Fmoc-Asn (Trt)-OH; Fmoc-Gly-OH; Fmoc-Leu-OH, Fmoc-Asn (Trt)-OH, Fmoc-Thr (tBu)-OH; Fmoc-Gly-OH; Fmoc-Val-OH, Fmoc-Asp (tBu)-OH, Fmoc-Leu-OH; Fmoc-Arg (Pbf)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH, Fmoc-Ser (tBu)-OH; Fmoc-Ile-OH, Fmoc-Pro-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 21

The preparation of modified anti-SIV peptide

In the present embodiment, synthetic and modify SEQ ID NO:64 according to following synthetic route for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:64 shows antiviral activity to simian immunodeficiency virus (SIV) as the crude product peptide.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH; Fmoc-Lys (Boc)-OH; Fmoc-Gln (Trt)-OH; Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH; Fmoc-Tyr (tBu)-OH, Fmoc-Met-OH, Fmoc-Asn (Trt)-OH; Fmoc-Lys (Boc)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Ile-OH; Fmoc-Gln (Trt)-OH, Fmoc-Ala-OH, Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Ala-OH; Fmoc-Thr (tBu)-OH; Fmoc-Ile-OH, Fmoc-Asn (Trt)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH, Fmoc-Phe-OH, Fmoc-Asp (tBu)-OH; Fmoc-Val-OH; Fmoc-Lys (Boc)-OH, Fmoc-Arg (Pbf)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Trp (Boc)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Trp (Boc)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mLCHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 22

The preparation of modified anti-SIV peptide

In the present embodiment, synthetic and modify SEQ ID NO:65 according to following synthetic route for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:65 shows antiviral activity to simian immunodeficiency virus (SIV) as the crude product peptide.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH; Fmoc-Leu-OH; Fmoc-Lys (Boc)-OH; Fmoc-Gln (Trt)-OH; Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH, Fmoc-Tyr (tBu)-OH, Fmoc-Met-OH; Fmoc-Asn (Trt)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Gln (Trt)-OH; Fmoc-Ile-OH, Fmoc-Gln (Trt)-OH, Fmoc-Ala-OH; Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH; Fmoc-Ala-OH; Fmoc-Thr (tBu)-OH, Fmoc-Ile-OH, Fmoc-Asn (Trt)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH, Fmoc-Phe-OH; Fmoc-Asp (tBu)-OH; Fmoc-Val-OH, Fmoc-Lys (Boc)-OH, Fmoc-Arg (Pbf)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Trp (Boc)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mLCHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 23

The preparation of modified anti-SIV peptide

In the present embodiment, synthetic and modify SEQ ID NO:66 according to following synthetic route for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:66 shows antiviral activity to simian immunodeficiency virus (SIV) as the crude product peptide.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Asn (Trt)-OH; Fmoc-Leu-OH; Fmoc-Lys (Boc)-OH; Fmoc-Gln (Trt)-OH; Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH; Fmoc-Tyr (tBu)-OH, Fmoc-Met-OH, Fmoc-Asn (Trt)-OH; Fmoc-Lys (Boc)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Ile-OH; Fmoc-Gln (Trt)-OH, Fmoc-Ala-OH, Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Ala-OH; Fmoc-Thr (tBu)-OH; Fmoc-Ile-OH, Fmoc-Asn (Trt)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH, Fmoc-Phe-OH, Fmoc-Asp (tBu)-OH; Fmoc-Val-OH; Fmoc-Lys (Boc)-OH, Fmoc-Arg (Pbf)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Trp (Boc)-OH, Fmoc-Glu (tBu)-OH Boc-Lys (Aloc)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mLCHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 24

The preparation of modified anti-SIV peptide

In the present embodiment, synthetic and modify SEQ ID NO:67 according to following synthetic route for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:67 shows antiviral activity to simian immunodeficiency virus (SIV) as the crude product peptide.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH; Fmoc-Ser (tBu)-OH; Fmoc-Asn (Trt)-OH; Fmoc-Leu-OH; Fmoc-Lys (Boc)-OH; Fmoc-Gln (Trt)-OH, Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH; Fmoc-Tyr (tBu)-OH; Fmoc-Met-OH, Fmoc-Asn (Trt)-OH, Fmoc-Lys (Boc)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Gln (Trt)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Ile-OH; Fmoc-Gln (Trt)-OH; Fmoc-Ala-OH, Fmoc-Glu (tBu)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH; Fmoc-Leu-OH, Fmoc-Ala-OH, Fmoc-Thr (tBu)-OH; Fmoc-Ile-OH; Fmoc-Asn (Trt)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH; Fmoc-Phe-OH, Fmoc-Asp (tBu)-OH, Fmoc-Val-OH; Fmoc-Lys (Boc)-OH; Fmoc-Arg (Pbf)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Trp (Boc)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mLCHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 25

The preparation of modified anti-SIV peptide

In the present embodiment, synthetic and modify SEQ ID NO:68 according to following synthetic route for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:68 shows antiviral activity to simian immunodeficiency virus (SIV) as the crude product peptide.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Trp (Boc)-OH; Fmoc-Ser (tBu)-OH; Fmoc-Asn (Trt)-OH; Fmoc-Leu-OH; Fmoc-Lys (Boc)-OH; Fmoc-Gln (Trt)-OH; Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH, Fmoc-Tyr (tBu)-OH; Fmoc-Met-OH; Fmoc-Asn (Trt)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Gln (Trt)-OH; Fmoc-Gln (Trt)-OH, Fmoc-Ile-OH, Fmoc-Gln (Trt)-OH; Fmoc-Ala-OH; Fmoc-Glu (tBu)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH; Fmoc-Leu-OH; Fmoc-Ala-OH, Fmoc-Thr (tBu)-OH, Fmoc-Ile-OH; Fmoc-Asn (Trt)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH; Fmoc-Phe-OH; Fmoc-Asp (tBu)-OH, Fmoc-Val-OH, Fmoc-Lys (Boc)-OH; Fmoc-Arg (Pbf)-OH, Fmoc-Glu (tBu)-OH Boc-Lys (Aloc)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mLCHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 26

The preparation of modified anti-SIV peptide

In the present embodiment, synthetic and modify SEQ ID NO:69 according to following synthetic route for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:69 shows antiviral activity to simian immunodeficiency virus (SIV) as the crude product peptide.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH; Fmoc-Asp (tBu)-OH; Fmoc-Trp (Boc)-OH; Fmoc-Ser (tBu)-OH; Fmoc-Asn (Trt)-OH; Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH, Fmoc-Tyr (tBu)-OH, Fmoc-Met-OH; Fmoc-Asn (Trt)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Gln (Trt)-OH; Fmoc-Ile-OH, Fmoc-Gln (Trt)-OH, Fmoc-Ala-OH; Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH; Fmoc-Ala-OH; Fmoc-Thr (tBu)-OH, Fmoc-Ile-OH, Fmoc-Asn (Trt)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH, Fmoc-Phe-OH; Fmoc-Asp (tBu)-OH; Fmoc-Val-OH, Fmoc-Lys (Boc)-OH, Fmoc-Arg (Pbf)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 27

The preparation of modified anti-SIV peptide

In the present embodiment, synthetic and modify SEQ ID NO:70 according to following synthetic route for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:70 shows antiviral activity to simian immunodeficiency virus (SIV) as the crude product peptide.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Val-OH; Fmoc-Asp (tBu)-OH; Fmoc-Trp (Boc)-OH; Fmoc-Ser (tBu)-OH; Fmoc-Asn (Trt)-OH; Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH, Fmoc-Tyr (tBu)-OH, Fmoc-Met-OH; Fmoc-Asn (Trt)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Gln (Trt)-OH; Fmoc-Ile-OH, Fmoc-Gln (Trt)-OH, Fmoc-Ala-OH; Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH; Fmoc-Ala-OH; Fmoc-Thr (tBu)-OH, Fmoc-Ile-OH, Fmoc-Asn (Trt)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH, Fmoc-Phe-OH; Fmoc-Asp (tBu)-OH; Fmoc-Val-OH, Fmoc-Lys (Boc)-OH, Boc-Lys (Aloc)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10u phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 28

The preparation of modified anti-SIV peptide

In the present embodiment, synthetic and modify SEQ ID NO:71 according to following synthetic route for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:71 shows antiviral activity to simian immunodeficiency virus (SIV) as the crude product peptide.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH, Fmoc-Phe-OH, Fmoc-Val-OH; Fmoc-Asp (tBu)-OH, Fmoc-Trp (Boc)-OH, Fmoc-Ser (tBu)-OH; Fmoc-Asn (Trt)-OH, Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH; Fmoc-Gln (Trt)-OH, Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH; Fmoc-Tyr (tBu)-OH, Fmoc-Met-OH, Fmoc-Asn (Trt)-OH; Fmoc-Lys (Boc)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Gln (Trt)-OH, Fmoc-Ile-OH, Fmoc-Gln (Trt)-OH; Fmoc-Ala-OH, Fmoc-Glu (tBu)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH; Fmoc-Leu-OH, Fmoc-Ala-OH, Fmoc-Thr (tBu)-OH; Fmoc-Ile-OH; Fmoc-Asn (Trt)-OH, Fmoc-Glu (tBu)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH; Fmoc-Phe-OH, Fmoc-Asp (tBu)-OH, Fmoc-Val-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mLCHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 29

The preparation of modified anti-SIV peptide

In the present embodiment, synthetic and modify SEQ ID NO:72 according to following synthetic route for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:72 shows antiviral activity to simian immunodeficiency virus (SIV) as the crude product peptide.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH, Fmoc-Gly-OH, Fmoc-Phe-OH; Fmoc-Val-OH, Fmoc-Asp (tBu)-OH, Fmoc-Trp (Boc)-OH; Fmoc-Ser (tBu)-OH, Fmoc-Asn (Trt)-OH, Fmoc-Leu-OH; Fmoc-Lys (Boc)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Leu-OH; Fmoc-Glu (tBu)-OH, Fmoc-Tyr (tBu)-OH, Fmoc-Met-OH; Fmoc-Asn (Trt)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Gln (Trt)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Ile-OH; Fmoc-Gln (Trt)-OH, Fmoc-Ala-OH, Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Ala-OH; Fmoc-Thr (tBu)-OH; Fmoc-Ile-OH, Fmoc-Asn (Trt)-OH, Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Leu-OH, Fmoc-Phe-OH, Fmoc-Asp (tBu)-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Embodiment 30

The preparation of modified anti-SIV peptide

In the present embodiment, synthetic and modify SEQ ID NO:73 according to following synthetic route for containing linker and maleimide base group.As United States Patent (USP) 6,013,236 and 6,020,459 is described, and SEQ ID NO:73 shows antiviral activity to simian immunodeficiency virus (SIV) as the crude product peptide.

The solid-phase peptide of the modified peptide on the 100 μ mole scales is synthetic to be to adopt synthetic, collaborative peptide synthesizer of manual solid-phase and Fmoc protection Rink acid amides mbha resin to carry out.The amino-acid sequence of following protection is added resin: Fmoc-Lys (Aloc)-OH, Fmoc-Asn (Trt)-OH, Fmoc-Gly-OH; Fmoc-Phe-OH, Fmoc-Val-OH, Fmoc-Asp (tBu)-OH; Fmoc-Trp (Boc)-OH, Fmoc-Ser (tBu)-OH, Fmoc-Asn (Trt)-OH; Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Leu-OH, Fmoc-Glu (tBu)-OH, Fmoc-Tyr (tBu)-OH; Fmoc-Met-OH, Fmoc-Asn (Trt)-OH, Fmoc-Lys (Boc)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Gln (Trt)-OH; Fmoc-Ile-OH, Fmoc-Gln (Trt)-OH, Fmoc-Ala-OH; Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH; Fmoc-Ala-OH; Fmoc-Thr (tBu)-OH, Fmoc-Ile-OH, Fmoc-Asn (Trt)-OH; Fmoc-Glu (tBu)-OH; Fmoc-Glu (tBu)-OH, Fmoc-Leu-OH, Fmoc-Phe-OH.They are dissolved in N, dinethylformamide (DMF) and according to sequence with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU) and diisopropylethylamine (DIEA) activate.With being present in N, 20% (V/V) piperidine solution in the dinethylformamide (DMF) was carried out (step 1) that removes of Fmoc protecting group in 20 minutes.By being dissolved in 5mL CHCl with 3 equivalents ₃: the Pd (PPh among NMM: the HOAc (18: 1: 0.5) ₃) ₄The solution-treated resin was manually finished the selectivity deprotection (step 2) of Lys (Aloc) base in 2 hours.Resin CHCl subsequently ₃(6 * 5mL), 20% be present in HOAc among the DCM (6 * 5mL), DCM (6 * 5mL) and DMF (6 * 5mL) washings.After this synthetic automatically once more so that add 3-dimaleoyl imino propionic acid (step 3).Between each coupling, resin N, dinethylformamide (DMF) washing 3 times is also used washed with isopropyl alcohol 3 times.Take off peptide with 85%TFA/5%TIS/5% thioanisole and 5% phenol from resin, use dry ice refrigerative Et subsequently ₂O precipitates (step 4).Product is by preparing the preparation binary HPLC system purifying of reversed-phase HPLC, use Varian (Rainin): the 30-55%B (H that contains 0.045%TFA in about 180 minutes ₂O (A) and contain the CH of 0.045%TFA ₃CN (B)) gradient elution to carry out in 9.5mL/ minute; Use the UV detector (Varian Dynamax UVD II) of Phenomenex Luna 10 μ phenyl-hexyl 21mm * 25cm post and λ 214 and 254nm, the modified peptide that obtains expecting (being DAC) is measured purity＞95% of peptide by RP-HPLC.

Though described and exemplified some embodiment of the present invention, one skilled in the art should understand the qualification that the present invention is not subjected to these embodiment particular contents, and are limited by the accompanying claims.

Table 2

Adopt the single-letter amino acid coding of table 1

Table 3

Adopt the single-letter amino acid coding of table 1

Table 4

Adopt the single-letter amino acid coding of table 1

Table 5

Adopt the single-letter amino acid coding of table 1

Table 6

Adopt the single-letter amino acid coding of table 1

Table 7

Adopt the single-letter amino acid coding of table 1

Table 8

Adopt the single-letter amino acid coding of table 1

Table 9

Adopt the single-letter amino acid coding of table 1

Table 10

Adopt the single-letter amino acid coding of table 1

Table 11

Adopt the single-letter amino acid coding of table 1

Table 12

Adopt the single-letter amino acid coding of table 1

Table 13

Adopt the single-letter amino acid coding of table 1

Table 14

Adopt the single-letter amino acid coding of table 1

Table 15

Adopt the single-letter amino acid coding of table 1

Table 16

Adopt the single-letter amino acid coding of table 1

Table 17

Adopt the single-letter amino acid coding of table 1

Table 18

Adopt the single-letter amino acid coding of table 1

Table 19

Adopt the single-letter amino acid coding of table 1

Sequence table

＜110〉Conjuchem Inc.

＜120〉long lasting fusion peptide inhibitor of virus infection

<130>SPI081664-01

<140>

<141>

<150>US?60/134,406

<151>1999-05-17

<150>US?60/153,406

<151>1999-09-10

<160>86

<170>PatentIn?Ver.2.1

<210>1

<211>36

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>1

Tyr?Thr?Ser?Leu?Ile?His?Ser?Leu?Ile?Glu?Glu?Ser?Gln?Asn?Gln?Gln

1 5 10 15

Glu?Lys?Asn?Glu?Gln?Glu?Leu?Leu?Glu?Leu?Asp?Lys?Trp?Ala?Ser?Leu

20 25 30

Trp?Asn?Trp?Phe

35

<210>2

<211>38

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>2

Asn?Asn?Leu?Leu?Arg?Ala?Ile?Glu?Ala?Gln?Gln?His?Leu?Leu?Gln?Leu

1 5 10 15

Thr?Val?Trp?Gln?Ile?Lys?Gln?Leu?Gln?Ala?Arg?Ile?Leu?Ala?Val?Glu

20 25 30

Arg?Tyr?Leu?Lys?Asp?Gln

35

<210>3

<211>36

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>3

Tyr?Thr?Asn?Thr?Ile?Tyr?Thr?Leu?Leu?Glu?Glu?Ser?Gln?Asn?Gln?Gln

1 5 10 15

Glu?Lys?Asn?Glu?Gln?Glu?Leu?Leu?Glu?Leu?Asp?Lys?Trp?Ala?Ser?Leu

20 25 30

Trp?Asn?Trp?Phe

35

<210>4

<211>36

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>4

Tyr?Thr?Gly?Ile?Ile?Tyr?Asn?Leu?Leu?Glu?Glu?Ser?Gln?Asn?Gln?Gln

1 5 10 15

Glu?Lys?Asn?Glu?Gln?Glu?Leu?Leu?Glu?Leu?Asp?Lys?Trp?Ala?Asn?Leu

20 25 30

Trp?Asn?Trp?Phe

35

<210>5

<211>36

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>5

Tyr?Thr?Ser?Leu?Ile?Tyr?Ser?Leu?Leu?Glu?Lys?Ser?Gln?Thr?Gln?Gln

1 5 10 15

Glu?Lys?Asn?Glu?Gln?Glu?Leu?Leu?Glu?Leu?Asp?Lys?Trp?Ala?Ser?Leu

20 25 30

Trp?Asn?Trp?Phe

35

<210>6

<211>36

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>6

Leu?Glu?Ala?Asn?Ile?Ser?Lys?Ser?Leu?Glu?Gln?Ala?Gln?Ile?Gln?Gln

1 5 10 15

Glu?Lys?Asn?Met?Tyr?Glu?Leu?Gln?Lys?Leu?Asn?Ser?Trp?Asp?Ile?Phe

20 25 30

Gly?Asn?Trp?Phe

35

<210>7

<211>36

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>7

Leu?Glu?Ala?Asn?Ile?Ser?Gln?Ser?Leu?Glu?Gln?Ala?Gln?Ile?Gln?Gln

1 5 10 15

Glu?Lys?Asn?Met?Tyr?Glu?Leu?Gln?Lys?Leu?Asn?Ser?Trp?Asp?Val?Phe

20 25 30

Thr?Asn?Trp?Leu

35

<210>8

<211>41

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>8

Cys?Gly?Gly?Asn?Asn?Leu?Leu?Arg?Ala?Ile?Glu?Ala?Gln?Gln?His?Leu

1 5 10 15

Leu?Gln?Leu?Thr?Val?Trp?Gly?Ile?Lys?Gln?Leu?Gln?Ala?Arg?Ile?Leu

20 25 30

Ala?Val?Glu?Arg?Tyr?Leu?Lys?Asp?Gln

35 40

<210>9

<211>38

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>9

Gln?Gln?Leu?Leu?Asp?Val?Val?Lys?Arg?Gln?Gln?Glu?Met?Leu?Arg?Leu

1 5 10 15

Thr?Val?Trp?Gly?Thr?Lys?Asn?Leu?Gln?Ala?Arg?Val?Thr?Ala?Ile?Glu

20 25 30

Lys?Tyr?Leu?Lys?Asp?Gln

35

<210>10

<211>46

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>10

Tyr?Thr?Ser?Val?Ile?Thr?Ile?Glu?Leu?Ser?Asn?Ile?Lys?Glu?Asn?Lys

1 5 10 15

Cys?Asn?Gly?Ala?Lys?Val?Lys?Leu?Ile?Lys?Gln?Glu?Leu?Asp?Lys?Tyr

20 25 30

Lys?Asn?Ala?Val?Thr?Glu?Leu?Gln?Leu?Leu?Met?Gln?Ser?Thr

35 40 45

<210>11

<211>54

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>11

Ala?Ser?Gly?Val?Ala?Val?Ser?Lys?Val?Leu?His?Leu?Glu?Gly?Glu?Val

1 5 10 15

Asn?Lys?Ile?Ala?Leu?Leu?Ser?Thr?Asn?Lys?Ala?Val?Val?Ser?Leu?Ser

20 25 30

Asn?Gly?Val?Ser?Val?Leu?Thr?Ser?Lys?Val?Leu?Asp?Leu?Lys?Asn?Tyr

35 40 45

Ile?Asp?Lys?Gln?Leu?Leu

50

<210>12

<211>53

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>12

Gly?Glu?Pro?Ile?Ile?Asn?Phe?Tyr?Asp?Pro?Leu?Val?Phe?Pro?Ser?Asp

1 5 10 15

Glu?Phe?Asp?Ala?Ser?Ile?Ser?Gln?Val?Asn?Glu?Lys?Ile?Asn?Gln?Ser

20 25 30

Leu?Ala?Phe?Ile?Arg?Lys?Ser?Asp?Glu?Leu?Leu?His?Asn?Val?Asn?Ala

35 40 45

Gly?Lys?Ser?Thr?Thr

50

<210>13

<211>48

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>13

Tyr?Thr?Ser?Val?Ile?Thr?Ile?Glu?Leu?Ser?Asn?Ile?Lys?Glu?Asn?Lys

1 5 10 15

Cys?Asn?Gly?Thr?Asp?Ala?Lys?Val?Lys?Leu?Ile?Lys?Gln?Glu?Leu?Asp

20 25 30

Lys?Tyr?Lys?Asn?Ala?Val?Thr?Glu?Leu?Gln?Leu?Leu?Met?Gln?Ser?Thr

35 40 45

<210>14

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>14

Tyr?Thr?Ser?Val?Ile?Thr?Ile?Glu?Leu?Ser?Asn?Ile?Lys?Glu?Asn?Lys

1 5 10 15

Cys?Asn?Gly?Asp?Ala?Lys?Val?Lys?Leu?Ile?Lys?Gln?Glu?Leu?Asp?Lys

20 25 30

Tyr?Lys

<210>15

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>15

Thr?Ser?Val?Ile?Thr?Ile?Glu?Leu?Ser?Asn?Ile?Lys?Glu?Asn?Lys?Cys

1 5 10 15

Asn?Gly?Asp?Ala?Lys?Val?Lys?Leu?Ile?Lys?Gln?Glu?Leu?Asp?Lys?Tyr

20 25 30

Lys?Asn

<210>16

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>16

Val?Ile?Thr?Ile?Glu?Leu?Ser?Asn?Ile?Lys?Glu?Asn?Lys?Cys?Asn?Gly

1 5 10 15

Asp?Ala?Lys?Val?Lys?Leu?Ile?Lys?Gln?Glu?Leu?Asp?Lys?Tyr?Lys?Asn

20 25 30

Ala?Val

<210>17

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>17

Val?Ile?Thr?Ile?Glu?Leu?Ser?Asn?Ile?Lys?Glu?Asn?Lys?Met?Asn?Gly

1 5 10 15

Asp?Ala?Lys?Val?Lys?Leu?Ile?Lys?Gln?Glu?Leu?Asp?Lys?Tyr?Lys?Asn

20 25 30

Ala?Val

<210>18

<211>33

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>18

Val?Ala?Val?Ser?Lys?Val?Leu?His?Leu?Glu?Gly?Glu?Val?Asn?Lys?Ile

1 5 10 15

Ala?Leu?Leu?Ser?Thr?Asn?Lys?Ala?Val?Val?Ser?Leu?Ser?Asn?Gly?Val

20 25 30

Ser

<210>19

<211>33

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>19

Ala?Val?Ser?Lys?Val?Leu?His?Leu?Glu?Gly?Glu?Val?Asn?Lys?Ile?Ala

1 5 10 15

Leu?Leu?Ser?Thr?Asn?Lys?Ala?Val?Val?Ser?Leu?Ser?Asn?Gly?Val?Ser

20 25 30

Val

<210>20

<211>33

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>20

Val?Ser?Lys?Val?Leu?His?Leu?Glu?Gly?Glu?Val?Asn?Lys?Ile?Ala?Leu

1 5 10 15

Leu?Ser?Thr?Asn?Lys?Ala?Val?Val?Ser?Leu?Ser?Asn?Gly?Val?Ser?Val

20 25 30

Leu

<210>21

<211>33

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>21

Ser?Lys?Val?Leu?His?Leu?Glu?Gly?Glu?Val?Asn?Lys?Ile?Ala?Leu?Leu

1 5 10 15

Ser?Thr?Asn?Lys?Ala?Val?Val?Ser?Leu?Ser?Asn?Gly?Val?Ser?Val?Leu

20 25 30

Thr

<210>22

<211>33

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>22

Lys?Val?Leu?His?Leu?Glu?Gly?Glu?Val?Asn?Lys?Ile?Ala?Leu?Leu?Ser

1 5 10 15

Thr?Asn?Lys?Ala?Val?Val?Ser?Leu?Ser?Asn?Gly?Val?Ser?Val?Leu?Thr

20 25 30

Ser

<210>23

<211>33

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>23

Leu?Glu?Gly?Glu?Val?Asn?Lys?Ile?Ala?Leu?Leu?Ser?Thr?Asn?Lys?Ala

1 5 10 15

Val?Val?Ser?Leu?Ser?Asn?Gly?Val?Ser?Val?Leu?Thr?Ser?Lys?Val?Leu

20 25 30

Asp

<210>24

<211>33

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>24

Gly?Glu?Val?Asn?Lys?Ile?Ala?Leu?Leu?Ser?Thr?Asn?Lys?Ala?Val?Val

1 5 10 15

Ser?Leu?Ser?Asn?Gly?Val?Ser?Val?Leu?Thr?Ser?Lys?Val?Leu?Asp?Leu

20 25 30

Lys

<210>25

<211>33

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>25

Glu?Val?Asn?Lys?Ile?Ala?Leu?Leu?Ser?Thr?Asn?Lys?Ala?Val?Val?Ser

1 5 10 15

Leu?Ser?Asn?Gly?Val?Ser?Val?Leu?Thr?Ser?Lys?Val?Leu?Asp?Leu?Lys

20 25 30

Asn

<210>26

<211>33

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>26

Val?Asn?Lys?Ile?Ala?Leu?Leu?Ser?Thr?Asn?Lys?Ala?Val?Val?Ser?Leu

1 5 10 15

Ser?Asn?Gly?Val?Ser?Val?Leu?Thr?Ser?Lys?Val?Leu?Asp?Leu?Lys?Asn

20 25 30

Tyr

<210>27

<211>33

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>27

Asn?Lys?Ile?Ala?Leu?Leu?Ser?Thr?Asn?Lys?Ala?Val?Val?Ser?Leu?Ser

1 5 10 15

Asn?Gly?Val?Ser?Val?Leu?Thr?Ser?Lys?Val?Leu?Asp?Leu?Lys?Asn?Tyr

20 25 30

Ile

<210>28

<211>33

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>28

Lys?Ile?Ala?Leu?Leu?Ser?Thr?Asn?Lys?Ala?Val?Val?Ser?Leu?Ser?Asn

1 5 10 15

Gly?Val?Ser?Val?Leu?Thr?Ser?Lys?Val?Leu?Asp?Leu?Lys?Asn?Tyr?Ile

20 25 30

Asp

<210>29

<211>33

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>29

Ile?Ala?Leu?Leu?Ser?Thr?Asn?Lys?Ala?Val?Val?Ser?Leu?Ser?Asn?Gly

1 5 10 15

Val?Ser?Val?Leu?Thr?Ser?Lys?Val?Leu?Asp?Leu?Lys?Asn?Tyr?Ile?Asp

20 25 30

Lys

<210>30

<211>33

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>30

Ala?Leu?Leu?Ser?Thr?Asn?Lys?Ala?Val?Val?Ser?Leu?Ser?Asn?Gly?Val

1 5 10 15

Ser?Val?Leu?Thr?Ser?Lys?Val?Leu?Asp?Leu?Lys?Asn?Tyr?Ile?Asp?Lys

20 25 30

Gln

<210>31

<211>70

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>31

Gly?Thr?Ile?Ala?Leu?Gly?Val?Ala?Thr?Ser?Ala?Gln?Ile?Thr?Ala?Ala

1 5 10 15

Val?Ala?Leu?Val?Glu?Ala?Lys?Gln?Ala?Arg?Ser?Asp?Ile?Glu?Lys?Leu

20 25 30

Lys?Glu?Ala?Ile?Arg?Asp?Thr?Asn?Lys?Ala?Val?Gln?Ser?Val?Gln?Ser

35 40 45

Ser?Ile?Gly?Asn?Leu?Ile?Val?Ala?Ile?Lys?Ser?Val?Gln?Asp?Tyr?Val

50 55 60

Asn?Lys?Glu?Ile?Val?Pro

65 70

<210>32

<211>56

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>32

Tyr?Thr?Pro?Asn?Asp?Ile?Thr?Leu?Asn?Asn?Ser?Val?Ala?Leu?Asp?Pro

1 5 10 15

Ile?Asp?Ile?Ser?Ile?Glu?Leu?Asn?Lys?Ala?Lys?Ser?Asp?Leu?Glu?Glu

20 25 30

Ser?Lys?Glu?Trp?Ile?Arg?Arg?Ser?Asn?Gln?Lys?Leu?Asp?Ser?Ile?Gly

35 40 45

Asn?Trp?His?Gln?Ser?Ser?Thr?Thr

50 55

<210>33

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>33

Thr?Leu?Asn?Asn?Ser?Val?Ala?Leu?Asp?Pro?Ile?Asp?Ile?Ser?Ile?Glu

1 5 10 15

Leu?Asn?Lys?Ala?Lys?Ser?Asp?Leu?Glu?Glu?Ser?Lys?Glu?Trp?Ile?Arg

20 25 30

Arg?Ser?Asn

35

<210>34

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>34

Leu?Asn?Asn?Ser?Val?Ala?Leu?Asp?Pro?Ile?Asp?Ile?Ser?Ile?Glu?Leu

1 5 10 15

Asn?Lys?Ala?Lys?Ser?Asp?Leu?Glu?Glu?Ser?Lys?Glu?Trp?Ile?Arg?Arg

20 25 30

Ser?Asn?Gln

35

<210>35

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>35

Asn?Asn?Ser?Val?Ala?Leu?Asp?Pro?Ile?Asp?Ile?Ser?Ile?Glu?Leu?Asn

1 5 10 15

Lys?Ala?Lys?Ser?Asp?Leu?Glu?Glu?Ser?Lys?Glu?Trp?Ile?Arg?Arg?Ser

20 25 30

Asn?Gln?Lys

35

<210>36

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>36

Asn?Ser?Val?Ala?Leu?Asp?Pro?Ile?Asp?Ile?Ser?Ile?Glu?Leu?Asn?Lys

1 5 10 15

Ala?Lys?Ser?Asp?Leu?Glu?Glu?Ser?Lys?Glu?Trp?Ile?Arg?Arg?Ser?Asn

20 25 30

Gln?Lys?Leu

35

<210>37

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>37

Ser?Val?Ala?Leu?Asp?Pro?Ile?Asp?Ile?Ser?Ile?Glu?Leu?Asn?Lys?Ala

1 5 10 15

Lys?Ser?Asp?Leu?Glu?Glu?Ser?Lys?Glu?Trp?Ile?Arg?Arg?Ser?Asn?Gln

20 25 30

Lys?Leu?Asp

35

<210>38

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>38

Val?Ala?Leu?Asp?Pro?Ile?Asp?Ile?Ser?Ile?Glu?Leu?Asn?Lys?Ala?Lys

1 5 10 15

Ser?Asp?Leu?Glu?Glu?Ser?Lys?Glu?Trp?Ile?Arg?Arg?Ser?Asn?Gln?Lys

20 25 30

Leu?Asp?Ser

35

<210>39

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>39

Ala?Leu?Asp?Pro?Ile?Asp?Ile?Ser?Ile?Glu?Leu?Asn?Lys?Ala?Lys?Ser

1 5 10 15

Asp?Leu?Glu?Glu?Ser?Lys?Glu?Trp?Ile?Arg?Arg?Ser?Asn?Gln?Lys?Leu

20 25 30

Asp?Ser?Ile

35

<210>40

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>40

Leu?Asp?Pro?Ile?Asp?Ile?Ser?Ile?Glu?Leu?Asn?Lys?Ala?Lys?Ser?Asp

1 5 10 15

Leu?Glu?Glu?Ser?Lys?Glu?Trp?Ile?Arg?Arg?Ser?Asn?Gln?Lys?Leu?Asp

20 25 30

Ser?Ile?Gly

35

<210>41

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>41

Asp?Pro?Ile?Asp?Ile?Ser?Ile?Glu?Leu?Asn?Lys?Ala?Lys?Ser?Asp?Leu

1 5 10 15

Glu?Glu?Ser?Lys?Glu?Trp?Ile?Arg?Arg?Ser?Asn?Gln?Lys?Leu?Asp?Ser

20 25 30

Ile?Gly?Asn

35

<210>42

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>42

Pro?Ile?Asp?Ile?Ser?Ile?Glu?Leu?Asn?Lys?Ala?Lys?Ser?Asp?Leu?Glu

1 5 10 15

Glu?Ser?Lys?Glu?Trp?Ile?Arg?Arg?Ser?Asn?Gln?Lys?Leu?Asp?Ser?Ile

20 25 30

Gly?Asn?Trp

35

<210>43

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>43

Ile?Asp?Ile?Ser?Ile?Glu?Leu?Asn?Lys?Ala?Lys?Ser?Asp?Leu?Glu?Glu

1 5 10 15

Ser?Lys?Glu?Trp?Ile?Arg?Arg?Ser?Asn?Gln?Lys?Leu?Asp?Ser?Ile?Gly

20 25 30

Asn?Trp?His

35

<210>44

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>44

Asp?Ile?Ser?Ile?Glu?Leu?Asn?Lys?Ala?Lys?Ser?Asp?Leu?Glu?Glu?Ser

1 5 10 15

Lys?Glu?Trp?Ile?Arg?Arg?Ser?Asn?Gln?Lys?Leu?Asp?Ser?Ile?Gly?Asn

20 25 30

Trp?His?Gln

35

<210>45

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>45

Ile?Ser?Ile?Glu?Leu?Asn?Lys?Ala?Lys?Ser?Asp?Leu?Glu?Glu?Ser?Lys

1 5 10 15

Glu?Trp?Ile?Arg?Arg?Ser?Asn?Gln?Lys?Leu?Asp?Ser?Ile?Gly?Asn?Trp

20 25 30

His?Gln?Ser

35

<210>46

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>46

Ser?Ile?Glu?Leu?Asn?Lys?Ala?Lys?Ser?Asp?Leu?Glu?Glu?Ser?Lys?Glu

1 5 10 15

Trp?Ile?Arg?Arg?Ser?Asn?Gln?Lys?Leu?Asp?Ser?Ile?Gly?Asn?Trp?His

20 25 30

Gln?Ser?Ser

35

<210>47

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>47

Ile?Glu?Leu?Asn?Lys?Ala?Lys?Ser?Asp?Leu?Glu?Glu?Ser?Lys?Glu?Trp

1 5 10 15

Ile?Arg?Arg?Ser?Asn?Gln?Lys?Leu?Asp?Ser?Ile?Gly?Asn?Trp?His?Gln

20 25 30

Ser?Ser?Thr

35

<210>48

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>48

Glu?Leu?Asn?Lys?Ala?Lys?Ser?Asp?Leu?Glu?Glu?Ser?Lys?Glu?Trp?Ile

1 5 10 15

ArgArg?Ser?Asn?Gln?Lys?Leu?Asp?Ser?Ile?Gly?Asn?Trp?His?Gln?Ser

20 25 30

Ser?Thr?Thr

35

<210>49

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>49

Thr?Ala?Ala?Val?Ala?Leu?Val?Glu?Ala?Lys?Gln?Ala?Arg?Ser?Asp?Ile

1 5 10 15

Glu?Lys?Leu?Lys?Glu?Ala?Ile?Arg?Asp?Thr?Asn?Lys?Ala?Val?Gln?Ser

20 25 30

Val?Gln?Ser

35

<210>50

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>50

Ala?Val?Ala?Leu?Val?Glu?Ala?Lys?Gln?Ala?Arg?Ser?Asp?Ile?Glu?Lys

1 5 10 15

Leu?Lys?Glu?Ala?Ile?Arg?Asp?Thr?Asn?Lys?Ala?Val?Gln?Ser?Val?Gln

20 25 30

Ser?Ser?Ile

35

<210>51

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>51

Leu?Val?Glu?Ala?Lys?Gln?Ala?Arg?Ser?Asp?Ile?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Ile?Arg?Asp?Thr?Asn?Lys?Ala?Val?Gln?Ser?Val?Gln?Ser?Ser?Ile

20 25 30

Gly?Asn?Leu

35

<210>52

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>52

Val?Glu?Ala?Lys?Gln?Ala?Arg?Ser?Asp?Ile?Glu?Lys?Leu?Lys?Glu?Ala

1 5 10 15

Ile?Arg?Asp?Thr?Asn?Lys?Ala?Val?Gln?Ser?Val?Gln?Ser?Ser?Ile?Gly

20 25 30

Asn?Leu?Ile

35

<210>53

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>53

Glu?Ala?Lys?Gln?Ala?Arg?Ser?Asp?Ile?Glu?Lys?Leu?Lys?Glu?Ala?Ile

1 5 10 15

Arg?Asp?Thr?Asn?Lys?Ala?Val?Gln?Ser?Val?Gln?Ser?Ser?Ile?Gly?Asn

20 25 30

Leu?Ile?Val

35

<210>54

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>54

Ala?Lys?Gln?Ala?Arg?Ser?Asp?Ile?Glu?Lys?Leu?Lys?Glu?Ala?Ile?Arg

1 5 10 15

Asp?Thr?Asn?Lys?Ala?Val?Gln?Ser?Val?Gln?Ser?Ser?Ile?Gly?Asn?Leu

20 25 30

Ile?Val?Ala

35

<210>55

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>55

Lys?Gln?Ala?Arg?Ser?Asp?Ile?Glu?Lys?Leu?Lys?Glu?Ala?Ile?Arg?Asp

1 5 10 15

Thr?Asn?Lys?Ala?Val?Gln?Ser?Val?Gln?Ser?Ser?Ile?Gly?Asn?Leu?Ile

20 25 30

Val?Ala?Ile

35

<210>56

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>56

Gln?Ala?Arg?Ser?Asp?Ile?Glu?Lys?Leu?Lys?Glu?Ala?Ile?Arg?Asp?Thr

1 5 10 15

Asn?Lys?Ala?Val?Gln?Ser?Val?Gln?Ser?Ser?Ile?Gly?Asn?Leu?Ile?Val

20 25 30

Ala?Ile?Lys

35

<210>57

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>57

Ala?Arg?Ser?Asp?Ile?Glu?Lys?Leu?Lys?Glu?Ala?Ile?Arg?Asp?Thr?Asn

1 5 10 15

Lys?Ala?Val?Gln?Ser?Val?Gln?Ser?Ser?Ile?Gly?Asn?Leu?Ile?Val?Ala

20 25 30

Ile?Lys?Ser

35

<210>58

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>58

Arg?Ser?Asp?Ile?Glu?Lys?Leu?Lys?Glu?Ala?Ile?Arg?Asp?Thr?Asn?Lys

1 5 10 15

Ala?Val?Gln?Ser?Val?Gln?Ser?Ser?Ile?Gly?Asn?Leu?Ile?Val?Ala?Ile

20 25 30

Lys?Ser?Val

35

<210>59

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>59

Ser?Asp?Ile?Glu?Lys?Leu?Lys?Glu?Ala?Ile?Arg?Asp?Thr?Asn?Lys?Ala

1 5 10 15

Val?Gln?Ser?Val?Gln?Ser?Ser?Ile?Gly?Asn?Leu?Ile?Val?Ala?Ile?Lys

20 25 30

Ser?Val?Gln

35

<210>60

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>60

Lys?Leu?Lys?Glu?Ala?Ile?Arg?Asp?Thr?Asn?Lys?Ala?Val?Gln?Ser?Val

1 5 10 15

Gln?Ser?Ser?Ile?Gly?Asn?Leu?Ile?Val?Ala?Ile?Lys?Ser?Val?Gln?Asp

20 25 30

Tyr?Val?Asn

35

<210>61

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>61

Leu?Lys?Glu?Ala?Ile?Arg?Asp?Thr?Asn?Lys?Ala?Val?Gln?Ser?Val?Gln

1 5 10 15

Ser?Ser?Ile?Gly?Asn?Leu?Ile?Val?Ala?Ile?Lys?Ser?Val?Gln?Asp?Tyr

20 25 30

Val?Asn?Lys

35

<210>62

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>62

Ala?Ile?Arg?Asp?Thr?Asn?Lys?Ala?Val?Gln?Ser?Val?Gln?Ser?Ser?Ile

1 5 10 15

Gly?Asn?Leu?Ile?Val?Ala?Ile?Lys?Ser?Val?Gln?Asp?Tyr?Val?Asn?Lys

20 25 30

Glu?Ile?Val

35

<210>63

<211>47

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>63

Thr?Trp?Gln?Glu?Trp?Glu?Arg?Lys?Val?Asp?Phe?Leu?Glu?Glu?Asn?Ile

1 5 10 15

Thr?Ala?Leu?Leu?Glu?Glu?Ala?Gln?Ile?Gln?Gln?Glu?Lys?Asn?Met?Tyr

20 25 30

Glu?Leu?Gln?Lys?Leu?Asn?Ser?Trp?Asp?Val?Phe?Gly?Asn?Trp?Phe

35 40 45

<210>64

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>64

Trp?Gln?Glu?Trp?Glu?Arg?Lys?Val?Asp?Phe?Leu?Glu?Glu?Asn?Ile?Thr

1 5 10 15

Ala?Leu?Leu?Glu?Glu?Ala?Gln?Ile?Gln?Gln?Glu?Lys?Ash?Met?Tyr?Glu

20 25 30

Leu?Gln?Lys

35

<210>65

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>65

Gln?Glu?Trp?Glu?Arg?Lys?Val?Asp?Phe?Leu?Glu?Glu?Asn?Ile?Thr?Ala

1 5 10 15

Leu?Leu?Glu?Glu?Ala?Gln?Ile?Gln?Gln?Glu?Lys?Asn?Met?Tyr?Glu?Leu

20 25 30

Gln?Lys?Leu

35

<210>66

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>66

Glu?Trp?Glu?Arg?Lys?Val?Asp?Phe?Leu?Glu?Glu?Asn?Ile?Thr?Ala?Leu

1 5 10 15

Leu?Glu?Glu?Ala?Gln?Ile?Gln?Gln?Glu?Lys?Asn?Met?Tyr?Glu?Leu?Gln

20 25 30

Lys?Leu?Asn

35

<210>67

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>67

Trp?Glu?Arg?Lys?Val?Asp?Phe?Leu?Glu?Glu?Asn?Ile?Thr?Ala?Leu?Leu

1 5 10 15

Glu?Glu?Ala?Gln?Ile?Gln?Gln?Glu?Lys?Asn?Met?Tyr?Glu?Leu?Gln?Lys

20 25 30

Leu?Asn?Ser

35

<210>68

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>68

Glu?Arg?Lys?Val?Asp?Phe?Leu?Glu?Glu?Asn?Ile?Thr?Ala?Leu?Leu?Glu

1 5 10 15

Glu?Ala?Gln?Ile?Gln?Gln?Glu?Lys?Asn?Met?Tyr?Glu?Leu?Gln?Lys?Leu

20 25 30

Asn?Ser?Trp

35

<210>69

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>69

Arg?Lys?Val?Asp?Phe?Leu?Glu?Glu?Asn?Ile?Thr?Ala?Leu?Leu?Glu?Glu

1 5 10 15

Ala?Gln?Ile?Gln?Gln?Glu?Lys?Asn?Met?Tyr?Glu?Leu?Gln?Lys?Leu?Asn

20 25 30

Ser?Trp?Asp

35

<210>70

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>70

Lys?Val?Asp?Phe?Leu?Glu?Glu?Asn?Ile?Thr?Ala?Leu?Leu?Glu?Glu?Ala

1 5 10 15

Gln?Ile?Gln?Gln?Glu?Lys?Asn?Met?Tyr?Glu?Leu?Gln?Lys?Leu?Asn?Ser

20 25 30

Trp?Asp?Val

35

<210>71

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>71

Val?Asp?Phe?Leu?Glu?Glu?Asn?Ile?Thr?Ala?Leu?Leu?Glu?Glu?Ala?Gln

1 5 10 15

Ile?Gln?Gln?Glu?Lys?Asn?Met?Tyr?Glu?Leu?Gln?Lys?Leu?Asn?Ser?Trp

20 25 30

Asp?Val?Phe

35

<210>72

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>72

Asp?Phe?Leu?Glu?Glu?Asn?Ile?Thr?Ala?Leu?Leu?Glu?Glu?Ala?Gln?Ile

1 5 10 15

Gln?Gln?Glu?Lys?Asn?Met?Tyr?Glu?Leu?Gln?Lys?Leu?Asn?Ser?Trp?Asp

20 25 30

Val?Phe?Gly

35

<210>73

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>73

Phe?Leu?Glu?Glu?Asn?Ile?Thr?Ala?Leu?Leu?Glu?Glu?Ala?Gln?Ile?Gln

1 5 10 15

Gln?Glu?Lys?Asn?Met?Tyr?Glu?Leu?Gln?Lys?Leu?Asn?Ser?Trp?Asp?Val

20 25 30

Phe?Gly?Asn

35

<210>74

<211>35

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>74

Pro?Asp?Ala?Val?Tyr?Leu?His?Arg?Ile?Asp?Leu?Gly?Pro?Pro?Ile?Ser

1 5 10 15

Leu?Glu?Arg?Leu?Asp?Val?Gly?Thr?Asn?Leu?Gly?Asn?Ala?Ile?Ala?Lys

20 25 30

Leu?Glu?Asp

35

<210>75

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>75

Leu?Glu?Arg?Leu?Asp?Val?Gly?Thr?Asn?Leu?Gly?Asn?Ala?Ile?Ala?Lys

1 5 10 15

Leu?Glu?Ala?Lys?Glu?Leu?Leu?Glu?Ser?Ser?Asp?Gln?Ile?Leu?Arg?Ser

20 25 30

Met?Lys

<210>76

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>76

Leu?His?Arg?Ile?Asp?Leu?Gly?Pro?Pro?Ile?Ser?Leu?Glu?Arg?Leu?Asp

1 5 10 15

Val?Gly?Thr?Asn?Leu?Gly?Asn?Ala?Ile?Ala?Lys?Leu?Glu?Ala?Lys?Glu

20 25 30

Leu?Leu

<210>77

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>77

His?Arg?Ile?Asp?Leu?Gly?Pro?Pro?Ile?Ser?Leu?Glu?Arg?Leu?Asp?Val

1 5 10 15

Gly?Thr?Asn?Leu?Gly?Asn?Ala?Ile?Ala?Lys?Leu?Glu?Ala?Lys?Glu?Leu

20 25 30

Leu?Glu

<210>78

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>78

Arg?Ile?Asp?Leu?Gly?Pro?Pro?Ile?Ser?Leu?Glu?Arg?Leu?Asp?Val?Gly

1 5 10 15

Thr?Asn?Leu?Gly?Asn?Ala?Ile?Ala?Lys?Leu?Glu?Ala?Lys?Glu?Leu?Leu

20 25 30

Glu?Ser

<210>79

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>79

Ile?Asp?Leu?Gly?Pro?Pro?Ile?Ser?Leu?Glu?Arg?Leu?Asp?Val?Gly?Thr

1 5 10 15

Asn?Leu?Gly?Asn?Ala?Ile?Ala?Lys?Leu?Glu?Ala?Lys?Glu?Leu?Leu?Glu

20 25 30

Ser?Ser

<210>80

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>80

Asp?Leu?Gly?Pro?Pro?Ile?Ser?Leu?Glu?Arg?Leu?Asp?Val?Gly?Thr?Asn

1 5 10 15

Leu?Gly?Asn?Ala?Ile?Ala?Lys?Leu?Glu?Ala?Lys?Glu?Leu?Leu?Glu?Ser

20 25 30

Ser?Asp

<210>81

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>81

Leu?Gly?Pro?Pro?Ile?Ser?Leu?Glu?Arg?Leu?Asp?Val?Gly?Thr?Asn?Leu

1 5 10 15

Gly?Asn?Ala?Ile?Ala?Lys?Leu?Glu?Ala?Lys?Glu?Leu?Leu?Glu?Ser?Ser

20 25 30

Asp?Gln

<210>82

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>82

Gly?Pro?Pro?Ile?Ser?Leu?Glu?Arg?Leu?Asp?Val?Gly?Thr?Asn?Leu?Gly

1 5 10 15

Asn?Ala?Ile?Ala?Lys?Leu?Glu?Ala?Lys?Glu?Leu?Leu?Glu?Ser?Ser?Asp

20 25 30

Gln?Ile

<210>83

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>83

Pro?Pro?Ile?Ser?Leu?Glu?Arg?Leu?Asp?Val?Gly?Thr?Asn?Leu?Gly?Asn

1 5 10 15

Ala?Ile?Ala?Lys?Leu?Glu?Ala?Lys?Glu?Leu?Leu?Glu?Ser?Ser?Asp?Gln

20 25 30

Ile?Leu

<210>84

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>84

Pro?Ile?Ser?Leu?Glu?Arg?Leu?Asp?Val?Gly?Thr?Asn?Leu?Gly?Asn?Ala

1 5 10 15

Ile?Ala?Lys?Leu?Glu?Ala?Lys?Glu?Leu?Leu?Glu?Ser?Ser?Asp?Gln?Ile

20 25 30

Leu?Arg

<210>85

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>85

Ser?Leu?Glu?Arg?Leu?Asp?Val?Gly?Thr?Asn?Leu?Gly?Asn?Ala?Ile?Ala

1 5 10 15

Lys?Leu?Glu?Ala?Lys?Glu?Leu?Leu?Glu?Ser?Ser?Asp?Gln?Ile?Leu?Arg

20 25 30

Ser?Met

<210>86

<211>34

<212>PRT

＜213〉artificial sequence

<220>

＜223〉artificial sequence description: synthetic peptide

<400>86

Leu?Glu?Arg?Leu?Asp?Val?Gly?Thr?Asn?Leu?Gly?Asn?Ala?Ile?Ala?Lys

1 5 10 15

Leu?Glu?Ala?Lys?Glu?Leu?Leu?Glu?Ser?Ser?Asp?Gln?Ile?Leu?Arg?Ser

20 25 30

Met?Lys

Claims (37)

1. the peptide of modified antiviral and anti-fusion comprises:

Peptide with antiviral and anti-fusion-activity, wherein said peptide is modified to have the connected group that contains maleimide, sulfydryl on this group and the albuminous halfcystine 34 has reactivity, and the wherein said group that contains maleimide is not connected with described peptide by linking group or be connected with described peptide by (2-amino) ethoxyacetic acid (AEA) or [2-(2-amino) oxyethyl group)] ethoxyacetic acid (AEEA) linking group.

2. the modified peptide of claim 1, wherein said peptide is DP178 or DP107, or has the peptide corresponding to the aminoacid sequence in human immunodeficiency virus (HIV) gp41 protein D P178 zone.

3. the peptide of modified antiviral and anti-fusion comprises:

The peptide that human immunodeficiency virus (HIV) is had antiviral and anti-fusion-activity, wherein said peptide is modified to have the connected group that contains maleimide, sulfydryl on this group and the albuminous halfcystine 34 has reactivity, and the wherein said group that contains maleimide is not connected with described peptide by linking group or be connected with described peptide by (2-amino) ethoxyacetic acid (AEA) or [2-(2-amino) oxyethyl group)] ethoxyacetic acid (AEEA) linking group.

4. each modified peptide among the claim 1-3, wherein said peptide is selected from the group of being made up of SEQID NO:1 to SEQ ID NO:9.

5. the modified peptide of claim 3, wherein said peptide is DP178 or DP107, or has the peptide corresponding to the aminoacid sequence in HIV gp41 protein D P178 zone.

6. the modified peptide of claim 1, wherein said peptide demonstrates antiviral and anti-fusion-activity to human respiratory syncytial virus (RSV).

7. the modified peptide of claim 6, wherein said peptide is selected from the group of being made up of SEQ ID NO:10 to SEQ ID NO:30.

8. the modified peptide of claim 6, wherein said peptide is selected from the group of being made up of SEQ ID NO:14 to SEQ ID NO:17 and SEQ ID NO:29.

9. the modified peptide of claim 1, wherein said peptide demonstrates antiviral and anti-fusion-activity to human parainfluenza virus (HPIV).

10. the modified peptide of claim 9, wherein said peptide is selected from the group of being made up of SEQ ID NO:31 to SEQ ID NO:62.

11. the modified peptide of claim 9, wherein said peptide are selected from the group of being made up of SEQ ID NO:35, SEQ ID NO:38 to SEQ ID NO:42, SEQ ID NO:52 and SEQ ID NO:58.

12. the modified peptide of claim 1, wherein said peptide demonstrates antiviral and anti-fusion-activity to Measles virus (MeV).

13. the modified peptide of claim 12, wherein said peptide are selected from the group of being made up of SEQ ID NO:74 to SEQ ID NO:86.

14. the modified peptide of claim 12, wherein said peptide are selected from the group of being made up of SEQ IDNO:77, SEQ ID NO:79, SEQ ID NO:81 and SEQ ID NO:84.

15. the modified peptide of claim 1, wherein said peptide demonstrates antiviral and anti-fusion-activity to simian immunodeficiency virus (SIV).

16. the modified peptide of claim 15, wherein said peptide are selected from the group of being made up of SEQ ID NO:63 to SEQ ID NO:73.

17. composition that is used to prevent and/or treat acquired immune deficiency syndrome (AIDS) (AIDS), described composition comprises the peptide that human immunodeficiency virus (HIV) is had antiviral and anti-fusion-activity, described peptide is modified with dimaleoyl imino, sulfydryl on described dimaleoyl imino and the albuminous halfcystine 34 has reactivity, and the wherein said group that contains maleimide is not connected with described peptide by linking group or is connected with described peptide by (2-amino) ethoxyacetic acid (AEA) or [2-(2-amino) oxyethyl group)] ethoxyacetic acid (AEEA) linking group.

18. the composition of claim 17, wherein said peptide is DP178 or DP107, or has the peptide corresponding to the aminoacid sequence in HIV gp41 protein D P178 zone.

19. one kind is used to prevent and/or treat the composition that human respiratory syncytial virus (RSV) infects, described composition comprises the peptide that RSV is had antiviral and anti-fusion-activity, described peptide is modified with dimaleoyl imino, sulfydryl on described maleimide base and the albuminous halfcystine 34 has reactivity, and the wherein said group that contains maleimide is not connected with described peptide by linking group or is connected with described peptide by (2-amino) ethoxyacetic acid (AEA) or [2-(2-amino) oxyethyl group)] ethoxyacetic acid (AEEA) linking group.

20. the composition of claim 19, wherein said peptide are selected from the group of being made up of SEQ ID NO:14 to SEQ ID NO:17 and SEQ ID NO:29.

21. one kind is used to prevent and/or treat the composition that human parainfluenza virus (HPIV) infects, described composition comprises the peptide that human parainfluenza virus (HPIV) is shown antiviral and anti-fusion-activity, described peptide is modified with dimaleoyl imino, sulfydryl on described dimaleoyl imino and the albuminous halfcystine 34 has reactivity, and the wherein said group that contains maleimide is not connected with described peptide by linking group or is connected with described peptide by (2-amino) ethoxyacetic acid (AEA) or [2-(2-amino) oxyethyl group)] ethoxyacetic acid (AEEA) linking group.

22. the composition of claim 21, wherein said peptide are selected from the group of being made up of SEQ ID NO:35, SEQ ID NO:38 to SEQ ID NO:42, SEQ ID NO:52 and SEQ ID NO:58.

23. one kind is used to prevent and/or treat the composition that Measles virus (MeV) infects, described composition comprises the peptide that Measles virus (MeV) is shown antiviral and anti-fusion-activity, described peptide is modified with dimaleoyl imino, sulfydryl on described dimaleoyl imino and the albuminous halfcystine 34 has reactivity, and the wherein said group that contains maleimide is not connected with described peptide by linking group or is connected with described peptide by (2-amino) ethoxyacetic acid (AEA) or [2-(2-amino) oxyethyl group)] ethoxyacetic acid (AEEA) linking group.

24. the composition of claim 23, wherein said peptide are selected from the group of being made up of SEQ ID NO:77, SEQ ID NO:79, SEQ ID NO:81 and SEQ ID NO:84.

25. claim 3, the application in the medicine of preparation handler's immunodeficiency virus (HIV) of 4 or 5 modified peptide.

26. claim 6, the application in the medicine of preparation handler's respiratory syncytial virus (RSV) of 7 or 8 modified peptide.

27. claim 9, the application in the medicine of preparation handler parainfluenza virus (HPIV) of 10 or 11 modified peptide.

28. claim 12,13 or 14 modified peptide are handled application in the medicine of Measles virus (MeV) in preparation.

29. the application of the modified peptide of claim 15 or 16 in the medicine of preparation processing simian immunodeficiency virus (SIV).

30. the modified peptide of claim 5, wherein said peptide corresponding to HIV gp41 protein D P178 zone comprises length range in addition at about 2 aminoacid sequences to about 50 amino-acid residues, and these aminoacid sequences are respectively corresponding to the gp41 zone of DP178 aminoacid sequence amino or carboxyl side.

31. each peptide in the claim 1,3,6,12,15 or 30, the group that wherein contains maleimide are dimaleoyl imino propionic acid (MPA) or γ-maleimide-butyramide (GMBA).

32. each composition in the claim 17,19,21 or 23, the group that wherein contains maleimide are dimaleoyl imino propionic acid (MPA) or γ-maleimide-butyramide (GMBA).

33. suppress or reduce the method for the film fusion between human immunodeficiency virus (HIV) and the cell, comprise the peptide that makes each modified anti-fusion among the HIV virus contact claim 1-5 or 30.

34. the method for the anti-peptide-albumin conjugate that merges of preparation comprises:

Make the peptide contact albumin of each modified anti-fusion in claim 1-5 or 30 form the anti-peptide-albumin conjugate that merges.

35. the method for claim 34, being prepared as of the peptide of wherein modified anti-fusion: peptide, the N-[γ-dimaleoyl imino butyryl acyloxy that will have the anti-fusion of free amine group] succinimide ester (GMBS) and triethylamine or the mixing of dimaleoyl imino propionic acid, formation contains the group of dimaleoyl imino at the free amine group place, thereby forms the peptide of modified anti-fusion.

36. each modified peptide in the claim 1,3,6,9,12 or 15, wherein said peptide reveals antiviral and anti-fusion-activity by virus-cytogamy hotlist that adjusting relates to coiled coil peptide structure.

37. each composition in the claim 17,19,21 or 23, wherein said peptide reveals antiviral and anti-fusion-activity by virus-cytogamy hotlist that adjusting relates to coiled coil peptide structure.