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WO1992022579A1 - PEPTIDES MIMANT LA gp120 - Google Patents

PEPTIDES MIMANT LA gp120 Download PDF

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Publication number
WO1992022579A1
WO1992022579A1 PCT/US1992/004972 US9204972W WO9222579A1 WO 1992022579 A1 WO1992022579 A1 WO 1992022579A1 US 9204972 W US9204972 W US 9204972W WO 9222579 A1 WO9222579 A1 WO 9222579A1
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Prior art keywords
peptide
amino acid
acid sequence
asn arg
ser
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PCT/US1992/004972
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English (en)
Inventor
Malcolm L. Gefter
Kenneth L. Melmon
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Immulogic Pharmaceutical Corporation
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Priority to EP92914211A priority Critical patent/EP0590057A1/fr
Publication of WO1992022579A1 publication Critical patent/WO1992022579A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/02—Libraries contained in or displayed by microorganisms, e.g. bacteria or animal cells; Libraries contained in or displayed by vectors, e.g. plasmids; Libraries containing only microorganisms or vectors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1037—Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • G01N33/56988—HIV or HTLV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • HIV Human immunodeficiency virus
  • gpl20 binds the cellular receptor of the virus, CD4, and cells expressing the envelope fuse with CD4-positive cells in culture. Fusion of infected and noninfected cells and infection by cell-free virions are important routes of HIV infection.
  • the amino acid sequence of the envelope is known to vary between different HIV isolates and the neutralizing antibodies elicited by the envelope of one HIV isolate are usually effective against only a subset of heterologous isolates. This observation suggests that such neutralizing antibodies are directed to a variable region of the envelope. This idea is supported by the fact that observation suggests that such neutralizing antibodies are directed to a variable region of the envelope. This idea is supported by the fact that neutralizing antibodies are elicited by an E. coli-produced recombinant fragment from the carboxyl-terminal region of gpl20, PB1, that contains 37% of gpl20 and is from a variable region of the envelope.
  • This fragment contains the dominant neutralizing epitopes (epitope being the basic element or smallest unit of recognition by a receptor such as an antibody), and a more complete understanding of the number, the location, and the potential role of antibodies to these epitopes in preventing viral infection may facilitate development of a subunit vaccine able to induce immunity to diverse HIV isolates.
  • This invention relates to peptides having a primary amino acid sequence which is unrelated to the primary gpl20 amino acid sequence and are, by themselves, or conjugated to an immunogenic carrier molecule, able to stimulate the production of anti-gpl20 antibodies.
  • the anti-gpl20 antibodies produced in response to peptides of the present invention are directed toward a region of the gpl20 molecule known as the V3-loop, which is known to contain important epitopes.
  • the peptides can be formulated in a physiologically acceptable carrier for use as a vaccine composition.
  • This composition can optionally include an adjuvant.
  • the peptide can be conjugated to an immunogenic carrier molecule to increase immunogenicity. In this context the peptide serve as an immunogen.
  • the invention also pertains to peptides which serve as antiviral agents and a method for interfering with viral propagation in an individual.
  • the method involves administering an effective amount of a therapeutic composition comprising a peptide in a physiological carrier.
  • the peptide has a primary amino acid sequence which is unrelated to the primary amino acid sequence of an essential protein of a virus (e.g., gpl20) and the peptide has the ability to interfere with a component involved in the function of the virus.
  • the peptide serves as an antiviral agent.
  • the invention in another aspect, pertains to a method for screening an epitope library with a non-isolate specific antibody.
  • Such antibodies typically do not bind antigen with high affinity.
  • Figure 1 is a schematic representation of results from a competition assay for V3-loop binding.
  • Figure 2 is a schematic representation of results from a competition assay for recombinant gpl20 binding.
  • Figure 3 is a schematic representation of results from a direct binding assay with serum from mice immunized with M306 G3.
  • Figure 4 is a schematic representation of results from a direct binding assay with serum from mice immunized with M306 G3 conjugated to an immunogenic carrier molecule.
  • Figure 5 is a schematic representation of results from a competition experiment in which V3-loop was attached to an ELISA plate and reacted with serum from mice immunized with M306 G3 conjugated to an immunogenic carrier molecule mixed with various peptides.
  • Figure 6 is a schematic representation of results from a competition experiment in which M306 G3 conjugated to bovine serum albumin was fixed to an ELISA plate and reacted with serum from mice immunized with M306 G3 and conjugated to an immunogenic carrier molecule mixed with various peptides.
  • Figure 7 is a schematic representation of results from a competition experiment in which V3-loop was fixed to an ELISA plate and the antibody was an affinity purified rat monoclonal known to be HIV neutralizing.
  • Figure 8 is a schematic representation of results from a competition experiment in which recombinant gpl20 was fixed to an ELISA plate and the antibody was an affinity purified rat monoclonal known to be HIV neutralizing.
  • One embodiment of subject invention is based on Applicants' discovery of peptides, unrelated in primary amino acid sequence to the primary amino acid sequence of gpl20, which stimulate the production of anti-gpl20 antibodies when administered in vivo to a mammal.
  • the primary amino acid sequence refers to the linear order of amino acid residues in a peptide or protein.
  • Peptides of the invention were identified initially by screening an epitope library with a rat monoclonal antibody known to be HIV neutralizing in an in vitro assay. Such a method is described, for example, by Scott and Smith (Science 24_iJ386-390 (1990)).
  • an epitope library is a vast mixture of filamentous phage clones, each displaying one peptide sequence on the virion surface.
  • the survey is accomplished using a binding protein (e.g., a monoclonal antibody) to affinity-purify phage that display tightly-binding peptides and propagating the purified phage in IL. coli.
  • the amino acid sequence of the peptides displayed on the phage are then determined by sequencing the corresponding coding region in the viral DNAs.
  • Peptides identified in this manner that are also capable of competing for antibody binding with the natural epitope are referred to herein as mimic peptides because they mimic the natural epitope, but in most instances are unrelated in primary sequence to the natural epitope.
  • the steps involved in the screening of an epitope library include providing an epitope library containing bacterial cells infected with fusion phage.
  • the fusion phage are then contacted with a biotinylated antibody under conditions appropriate for the binding of the antibody to peptides displayed on the surface of the bacterial cells infected by the fusion phage.
  • the mixture is then contacted with a streptavidin-coated substrate under conditions appropriate for the binding of biotin to streptavidin.
  • Non-specifically bound phage are removed with a wash step. Bound phage are eluted and used to infect I coli cells and peptide sequences binding to the rat monoclonal antibody were identified.
  • amino acid residues 1 through 4 and 11 through 14 were identical for all the peptides in the epitope library.
  • amino acid residues 1 through 4 and 11 through 14 are critical since amino acid residues 1 through 4 and 11 through 14 were part of the peptide that actually demonstrated function in the Exemplification.
  • Peptides having the same sequences as the peptides identified by screening the epitope library, except for the addition of amino acid 15 (alanine) were synthesized and assayed for the ability to stimulate an anti-gpl20 or anti-V3-loop response when administered to mice in vivo, as described in the Exemplification below.
  • Murine antisera was tested for reactivity in a direct binding assay as described in detail in the Exemplification section.
  • Three peptides were identified which stimulate a response which is cross-reactive with the natural HIV epitope.
  • the three peptides are 15 amino acid residues in length and can be represented generically as follows:
  • the symbols AAl, AA2 and AA3 represent amino acid residues which are not perfectly conserved in the three mimic peptides described herein.
  • the peptides of this invention include peptides of the generic formula set forth above wherein AAl, AA2 and AA3 represent any amino acid residue.
  • the peptides of the invention include fragments of the 15 amino acid sequence which have the desired properties discussed herein. More preferably, AAl and AA3 are selected from the group of amino acids consisting of serine, threonine, methionine and cysteine and AA2 is selected from the group consisting of lysine, arginine, histidine, serine and threonine. Due to the critical nature of the hexapeptide, the peptides of this invention can also be represented generically including the following sequence: AA1 Gin AA2 Asn Arg AA3
  • Peptides of the generic formula set forth above can be routinely synthesized by standard chemical techniques or by recombinant DNA methodology. Such techniques are well known in the art. Peptide synthesizers are available commercially and standard techniques have been described in numerous publications including, for example, Merrifield (J. Chem. Soc. 85:2149-2154 (1963)) and Hunkapillar et al. (Nature 310:105-111 (1984)). It is well known to one skilled in the art that the immunogenicity of small non-, or weakly-immunogenic molecules (haptens) can be increased by conjugating the weakly- immunogenic molecule to a carrier protein. Commonly used carrier proteins include keyhole limpet hemacyanin and bovine serum albumin. The peptides of this invention can be conjugated to a carrier molecule, using conventional methods, to enhance their immunogenic properties.
  • Another embodiment of the subject invention is based on Applicants' discovery of peptides, unrelated in primary amino acid sequence to the primary amino acid sequence of an essential protein of a virus (e.g., HIV gpl20), which have the ability to interfere with a component involved in the function of the virus.
  • these peptides have the ability by themselves, or conjugated to an immunogenic carrier molecule, to stimulate the production of antibodies specifically reactive with the essential viral protein when administered to a mammal such as the peptides set forth above which are mimic peptides for the V3 loop.
  • the amino acid sequences of the peptides set forth above represent only the essential portion required for immunogenic or anti-viral activity.
  • a peptide which is useful as an immunogen or anti-viral agent is not necessarily limited to a minimal sequence shown above, but must have at least a portion of an amino acid sequence shown above which provides immunogenic or anti-viral activity.
  • peptides which include modified amino acids are included within the scope of the invention provided that the modifications (e.g., conservative substitutions) do not adversely affect the functional characteristics of the peptides.
  • the mimic peptides can be used, for example, as immunogens to stimulate an anti-g ⁇ l20 immune response in an individual. Such a response could be protective in the sense that the individual to whom the peptides are administered may be immune to infection by the HIV virus.
  • the mimic peptides are formulated in a composition and administered according to conventional protocols.
  • Such a composition includes at least one mimic peptide, by itself or conjugated to an immunogenic carrier molecule, in an amount sufficient to stimulate an immune response in a host.
  • the mimic peptide is contained in a physiologically acceptable carrier.
  • Such carriers include, for example, fillers, non-toxic buffers, physiological saline solution, etc.
  • the composition can also include adjuvants, protease inhibitors, lymphokines, or compatible drugs where such a combination is seen as desirable or advantageous.
  • the role of the V3-loop has not been elucidated, it appears that the V3-loop has a viral function which leads to viral propagation.
  • Other pathogenic viruses contain proteins which have a viral function which lead to viral propagation.
  • Peptides which are unrelated in primary amino acid sequence to the primary amino acid sequence of an essential protein of a virus, preferably mimic peptides, can therefore be used in a therapeutic composition to interfere with viral replication in an individual.
  • the mimic peptides can be used as decoy molecules to inhibit the natural function of an essential viral protein (e.g., the HIV gpl20 protein) .
  • an effective amount of the mimic peptides to an infected or non-infected individual, it is reasonable to predict that their presence will interfere with viral replication thereby slowing or preventing the spread of the infection.
  • test peptides and test antibody were determined by ELISA assay.
  • Costar ELA/RIA plates were coated by overnight incubation with 50 ⁇ l of a solution containing the test peptides or controls. Following the overnight incubation, the plates were washed 3 times with 1 X Phosphate Buffered Saline (PBS) . The plates were then blocked with 1% Bovine Serum Albumin (BSA) essentially globulin free (Sigma Cat. #7638) at room temperature for about 1 hour or more.
  • BSA Bovine Serum Albumin
  • the blocker was removed and about 100 ⁇ l of affinity purified 1° antibody (or test serum) was added in a dilution series.
  • the dilutions were made in PBS-T/FCS (1 X PBS, 0.05% Tween-20, 2.5% Fetal Calf Serum (FCS) .
  • the serum was incubated on the plate for about 1 hour at room temperature. Following this incubation, the plates were washed 3 times with PBS-T/FCS. 2° antibody was then added (at about 100 ⁇ l per well) and incubated at room temperature for about 1 hour.
  • the 2° antibody is typically an anti-species antibody which is biotin labeled. This incubation is followed by 3 PBS-T/FCS washes.
  • a Streptavidin-HRP (horseradish peroxidase) conjugate (Southern Biotechnology Associates, Birmingham, AL) was added at an appropriate dilution (typically 1/10,000) in PBS-T/FCS and incubated at room temperature for about 1 hour. This step was followed by 3 washes with PBS-T/FCS.
  • the assay was developed using the TMB Microwell Substrate System (Kirkegaard & Perry Labs, Gaithersburg, MD) .
  • the optical density (OD) was read on an ELISA reader at 450 nanometers.
  • Competition experiments were carried out by mixing each test peptide and control peptide in solution with a test antibody and determining the effect (if any) of the peptide in solution on binding of the antibody to a peptide attached to the plate in an ELISA assay.
  • the protocol is essentially as described above in connection with the ELISA assay except that peptides (or peptide conjugates) to be tested for ability to compete with the fixed peptides are mixed with the antibody and incubated for about 30 minutes prior to the addition of the antibody to the ELISA plate.
  • V3-loop 50 ⁇ g/ml
  • non-cyclized V3-loop 50 ⁇ g/ml
  • recombinant gpl20 2 ⁇ g/ml
  • gpl20 2 ⁇ g/ml
  • Each of the 4 rat monoclonal antibodies was tested in the assay.
  • a murine control which was known to bind V3-loop was included as a positive control.
  • the results from this binding assay clearly showed that all four rat monoclonal antibodies react specifically with the cyclized and non-cyclized form of V3-loop.
  • Antibody 1G7 demonstrated the strongest reactivity.
  • Two of the antibodies were biotinylated (1G7 and 28G12) by conventional methods. Specifically, in separate 1.5 ml polypropylene tubes, 167 ⁇ l (500 ⁇ g) of 28G12 and 250 ⁇ l of 1G7 (500 ⁇ g) were added. To each tube was added 12 ⁇ l of a biotin solution and the tubes were incubated at room temperature, with agitation, for 2 hours. The biotin solution was prepared by dissolving 1-3 mgs NHS-LC Biotin (Pierce Cat. #21335; MW 556.59 daltons) in 456.6 ⁇ l distilled water/mg of reagents. The final concentration was 3.93 mM. The solution was made fresh each time the experiment was performed.
  • IM ethanalamine 500 ⁇ l was added (pH 9) and the mixture was incubated at room temperature for 2 hours. 20 ⁇ l of carrier ovalbumin (10 ⁇ g/ml was also added) and the biotinylated antibodies were diluted against 1 X TBS overnight. The concentration of each was determined to be about 400-500 ⁇ g/ml. The material was concentrated in a Centricon R 30-kilodalton ultrafilter (Amicon), and washed three times with 2ml TBS to remove unconjugated biotin.
  • UV-killed blocking phage 300 ⁇ l Of TBS/0.02% NaN3 and 40 ⁇ l (7.44 X 10 12 virions) UV-killed blocking phage was added directly to the Centricon R . The mixture was vortexed to thoroughly mix the blocking phage with the antibody solution, then reconcentrated. The blocking phage are meant to block antibodies that cross-react with the filamentous phage in general, apart from the foreign peptide they display. The retentate was collected in the conical cup by back-centrifugation, as described in the Centricon R instructions, and stored at 4°C.
  • Biotinylated antibodies prepared as described above, were used to screen an epitope library (Scott and Smith, Science 249:386-390 (1990)).
  • the epitope library was screened essentially as described by Scott and Smith (supra) .
  • peptides were discovered which specifically bind the HIV neutralizing rat monoclonal antibodies described above.
  • Four 15 amino acid length synthetic peptides were produced which contain the 14 amino acid peptide identified by screening the epitope library. These peptides are referred to herein as M303 G7, M305 G14, M306 G3 and M307 G10 and the sequences of these peptides are as follows:
  • the V3-loop sequence is: Gly Cys Thr Arg Pro Asn Asn Asn Thr Arg Lys Ser lie Arg lie Gin Arg Gly Pro Gly Arg Ala Phe Val Thr lie Gly Lys lie Gly Asn Met Arg Gin Ala His Cys Gly.
  • the cyclized form of the V3-loop is generated by joining the amino terminus and the carboxy terminus of the linear molecule.
  • M303 G7 Reactivity of M303 G7. M305 G14 and M306 G3 with rat monoclonal 28GI2
  • the M303 G7, M305 G14, M306 G3 and M307 G10 peptides were synthesized by conventional methods and tested for direct binding with rat monoclonal antibody 28G12 by the ELISA assay described above. Each of the four peptides was attached to separate wells of an ELISA plate. Appropriate controls were included such as an V3-loop positive control in an ELISA well and a BSA negative control in a separate well.
  • the antibody showed strong binding to the positive controls (V3-loop and recombinant gpl20) but showed binding to M303 G7, M305 G14, M306 G3 and M307 G10 at near background levels or slightly elevated. This may be an artifact of the small size of these peptides (15 amino acids as compared to 38 for V3-loop) .
  • test peptides were tested in the competition assay described above. Specifically, when three of the four test peptides were incubated with the 28G12 antibody in solution prior to incubation with plates coated with V3-loop or recombinant gpl20 a significant decrease in antibody binding, as evidenced by a decrease on the OD45 0 determination, was observed. When tested in this manner, the peptide M307 G10 was the only test peptide that did not result in a significant decrease in antibody binding.
  • This M307 G10 peptide had glycine as the ninth amino acid residue, whereas the other three test peptides, M303 G7, M305 G14 and M306 G3 all had arginine as the ninth amino acid residue.
  • the control peptides used were lambda repressor amino acid residues 12-26, said peptide having the sequence Leu Glu Asp Ala Arg Arg Leu Lys Ala lie Tyr Glu Lys Lys Lys (CONTROL in Figures 1-7) and a synthesized peptide, M304 P8, containing the sequence of a 14 amino acid peptide that did not bind biotinylated antibody during the screening of the epitope library, said M304 P8 peptide having the sequence Ala Asp Gly Ala lie Ser Asn Leu lie Ser Gly Ala Ala Gly Ala.
  • the results of this experiment are shown in Figures 1 and 2.
  • mice were immunized using conventional methods with each of the three mimic peptides.
  • the mice were primed on Day 0 with 50 ⁇ g per mouse of either M306 G3 in complete Freund's adjuvant (CFA) or M306 G3 conjugated to keyhole limpet hemacyanin (KLH) in CFA.
  • CFA complete Freund's adjuvant
  • KLH keyhole limpet hemacyanin
  • IFA incomplete Freund's adjuvant
  • Figure 3 represents an average value of assays performed after each bleed from 4 mice immunized with M306 G3.
  • the figure legend lists the antigen which was bound to the plate.
  • the control was bovine serum albumin (BSA).
  • BSA bovine serum albumin
  • immunization with M306 G3 mimic peptide led to the production of murine antibodies specifically reactive with both V3-loop and the M306 G3 peptide.
  • the immunized mice had never been exposed to KLH and therefore the reactivity seen in Figure 3, middle bar, is due to specific reactivity with the mimic peptide.
  • immunization with the mimic peptide results in the production of murine antibodies which are specifically reactive with the V3-loop peptide.
  • Figure 4 shows the results of a similar experiment in which M306 G3 conjugated to KLH was used to immunize 4 mice.
  • the data presented in Figure 4 represents an average value for the 4 mice.
  • the antigen which was fixed to the plate is shown in the legend.
  • KLH was not present on the plate and therefore the middle bar represents reactivity toward the mimic peptide specifically.
  • the antisera was reactive with V3-loop although a greater disparity was observed between the relative affinities of the antisera for V3-loop versus M306 G3 than was observed in the experiment described in the preceding paragraph.
  • FIG. 6 shows the results of competition experiments in which M306 G3 BSA was fixed to the ELISA plate and the serum was from a mouse immunized with M306 G3 KLH.
  • the M306 G3 peptide was able to compete effectively at antigen concentrations exceeding 10 ⁇ g/ml.
  • FIG. 7 shows the results of a competition experiment with this antibody using ELISA plates to which V3-loop had been attached. Again, M306 G3 mimic peptide showed competitive behavior at concentrations exceeding 10 ⁇ g/ml.
  • Figure 8 shows the results of a 28G12 competition experiment in which recombinant g ⁇ l20 was fixed to the ELISA plate. In this experiment, the inclusion of the M306 G3 peptide resulted in a substantial reduction in the OD determination indicating effective competition.

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Abstract

Cette invention concerne des peptides comprenant une séquence primaire d'aides aminés qui n'est pas liée à la gp120. Les peptides se caractérisent par leur aptitude à stimuler la production d'anticorps anti-gp120, plus particulièrement d'anticorps anti-boucle-V3 lorsqu'on les administre à un mammifère. Ces peptides se caractérisent également par leur capacité à entraver avec la fonction virale. On peut formuler lesdits peptides sous forme d'une composition vaccinale et les administrer pour stimuler la réponse immunitaire neutralisant le VIH-1.
PCT/US1992/004972 1991-06-19 1992-06-19 PEPTIDES MIMANT LA gp120 WO1992022579A1 (fr)

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US72906491A 1991-07-12 1991-07-12
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0280468A2 (fr) * 1987-02-19 1988-08-31 Nissin Shokuhin Kabushiki Kaisha Méthodes et matériaux de détection et de thérapie du VIH
EP0373070A1 (fr) * 1988-12-06 1990-06-13 Centre National De La Recherche Scientifique Procédé de préparation d'une librairie de déterminants antigéniques peptidiques, nouveaux peptides constitués par ou comportant les déterminants antigéniques obtenus et application des peptides, notamment au diagnostic

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0280468A2 (fr) * 1987-02-19 1988-08-31 Nissin Shokuhin Kabushiki Kaisha Méthodes et matériaux de détection et de thérapie du VIH
EP0373070A1 (fr) * 1988-12-06 1990-06-13 Centre National De La Recherche Scientifique Procédé de préparation d'une librairie de déterminants antigéniques peptidiques, nouveaux peptides constitués par ou comportant les déterminants antigéniques obtenus et application des peptides, notamment au diagnostic

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
GENE. vol. 71, 1988, AMSTERDAM NL pages 247 - 256 WRAY AND FISHER 'Cloning and nucleotide sequence of the Streptomyces coelicor gene encoding glutamine sythetase' underlined sequences in figure 2, page 251 *
GENE. vol. 99, no. 2, 23 April 1991, AMSTERDAM NL pages 261 - 265 Y.TSUNETSUGU-YOKOTA ET AL. 'Expression of an immunogenic region of HIV by a filamentous bacteriophage vector' page 265 paragraph (3) *
SCIENCE. vol. 249, 27 July 1990, LANCASTER, PA US pages 386 - 390 SCOTT AND SMITH 'Searching for peptide ligands with an epitope library' cited in the application *

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