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WO2007083806A1 - Peptide antigène à restriction hla-a2 dérivés du virus de l'hépatite c 2a - Google Patents

Peptide antigène à restriction hla-a2 dérivés du virus de l'hépatite c 2a Download PDF

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Publication number
WO2007083806A1
WO2007083806A1 PCT/JP2007/050966 JP2007050966W WO2007083806A1 WO 2007083806 A1 WO2007083806 A1 WO 2007083806A1 JP 2007050966 W JP2007050966 W JP 2007050966W WO 2007083806 A1 WO2007083806 A1 WO 2007083806A1
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Prior art keywords
peptide
seq
hcv
hcv2a
cells
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English (en)
Japanese (ja)
Inventor
Akira Yamada
Kyogo Itoh
Michio Sata
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Kurume University
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Kurume University
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • 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
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/24011—Flaviviridae
    • C12N2770/24211—Hepacivirus, e.g. hepatitis C virus, hepatitis G virus
    • C12N2770/24222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • the present invention relates to a composition useful for treatment of a hepatitis C virus (HCV) -related disease and diagnosis or progression prediction of an HCV-related disease.
  • HCV hepatitis C virus
  • HCV Hepatitis C virus
  • CTL cytotoxic T lymphocyte
  • HCV is a highly mutated virus, and at least six genotypes have been found in the world (Bukh, J “et al, Semin. Liver Dis. 1995. 15: 41-63). Of these, lb is the main genotype of HCV, and so much research has been conducted on these types, and several HLA-restricted epitopes derived from HCVlb have been used for peptide vaccines. It has been identified as a candidate (WO2005 / 028503, Kurok ohchi K, et al "J Hepatol. 2001; 34 (6): 930-935; Uno— Furuta S, et al., V accine.
  • HCV2a is the dominant genotype in China, Japan and Italy, little is known about HCV2a (Zein NN, Clin Microbiol Re v. 2000; 13 (2): 223-235 Huy TT, Abe K., Pediatr Int. 2004; 46 (2): 223-23 0).
  • an object of the present invention is to provide a molecule that is the basis for specific immunotherapy for hepatitis C virus (HCV) 2a infection.
  • Patent Document 1 WO2005 / 028503
  • Non-patent literature l Kurokohchi K, et al., J Hepatol. 2001; 34 (6): 930-935
  • Non-patent literature 2 Uno- Furuta S, et al, Vaccine. 2001; 19: 2190-2196
  • An object of the present invention is to provide a therapy for HCV-related diseases that can be applied to patients infected with HCV2a.
  • HCV hepatitis C virus
  • HLA human leukocyte antigen
  • the present invention provides a peptide having any of the following amino acid sequences:
  • VVLDSLDPMV SEQ ID NO: 7
  • NS5 2850-2858 WLGNIIQYA (SEQ ID NO: 8) NS5 3012-3021; RLLLLGLLLL (SEQ ID NO: 9)
  • peptides can be recognized by antibodies against hepatitis C virus.
  • the peptides of the present invention have any of the following sequences:
  • VVLDSLDPMV SEQ ID NO: 7
  • the peptide of the present invention can induce HLA-A2-restricted cytotoxic T cells.
  • the present invention provides a pharmaceutical composition for treating a patient having an HCV-related disease, comprising the above-described peptide of the present invention as an active ingredient.
  • the present invention also provides a composition for diagnosing HCV-related diseases or predicting the progression of HCV-related diseases, comprising the above-described peptide of the present invention.
  • HCV-related disease means a disease caused by infection with hepatitis C virus, such as acute and chronic hepatitis, cirrhosis, and hepatocellular carcinoma. included.
  • the progression of HCV-related disease is typically progression from chronic hepatitis to cirrhosis and further to cirrhotic liver cancer.
  • Figure 1 shows the cytotoxic activity of HCV2a peptide-specific CTLs derived from PBMC of patients infected with HCV2a.
  • FIG. 2 shows the cytotoxic activity of HCV2a peptide-specific CTL derived from healthy donors.
  • FIG. 3 shows the cytotoxic activity of HCV2a peptide-induced CTLs against cells transfected with HCV2aNS3-NS5.
  • the present invention provides an immunogenic HCV-derived peptide that can be used for the treatment of a patient infected with HCV2a.
  • the hepatitis C virus-derived peptide (HCV peptide) of the present invention includes a partial amino acid sequence of a protein derived from hepatitis C virus type 2a, and includes an HLA-binding motif compatible with the patient's HLA in the sequence. This peptide is It can be recognized by antibodies in the blood of patients infected with Luz.
  • the hepatitis C virus-derived peptide according to the present invention can induce CTL, and the CTL thus induced targets HCV-infected cells and attacks the cells.
  • peripheral blood mononuclear cells of HCV2a-infected patients are stimulated with an HCV2a-derived peptide having an HLA-A2 binding motif, and peptide-specific
  • HLA-A2-restricted cytotoxic activity was examined.
  • peptide-specific CTLs were effectively induced from PBMCs of 9 types of peptide strength HLA-A2-positive HCV2a-infected patients.
  • the percentage of patients who responded to the peptide varied with each peptide and was in the range of 13-67%.
  • HCV2aE2 432 441; SLHTGFLASL (SEQ ID NO: 4), HCV2aE 2 716-724; YIVRWEWVV (SEQ ID NO: 5) and HCV2aNS5 2251-2260;
  • WLDSLDPMV (SEQ ID NO: 7)) induced CTL at a particularly high frequency.
  • peptide-specific CD8 + T cells reactive to each peptide showed HLA-A2-restricted cytotoxic activity.
  • HCV2a2251-2260-reactive T cells had cytotoxic activity against target cells transfected with the HCV 2aNS3-NS5 gene.
  • HLA class I binding tumor antigen-derived peptides previously identified as capable of inducing tumor-reactive CTLs are also recognized by IgG (Ohkouchi S, et al. Tissue Antigens. 2002; 59: 259-272; Kawamoto N, et al, Tissue Antigens. 2003; 61: 352-361), in clinical trials, peptide vaccines are often known to induce IgG reactive to the peptide. (Noguchi M, et al., Prost ate. 2003; 57: 80-92; Tanaka S, et al., J Immunother. 2003; 26: 357-366).
  • HCV is known to cause base sequence mutations at high frequency in its genome.
  • GenBank database HC—J6, PH77C V-J6S, PJ6CF, JCH-1, MD2A—1, NDM228, JFH-1, and G2AK3
  • HCV2a576-584, HCV2a627-635, and HCV2al085-1094 were found to have amino acid mutations in 2, 3 or 1 of each of the 8 strains. That is, it is considered that a peptide having an amino acid sequence mutated from the amino acid sequence shown in SEQ ID NOs: 2-9 can induce a peptide-specific CTL against HCV having such a mutation, and therefore, within the scope of the present invention. is there.
  • 1, 2 or 3 amino acid residues may be conservatively substituted in the amino acid sequences shown in SEQ ID NOs: 2-9.
  • Conservative substitutions are those in which one amino acid is replaced with another amino acid with similar properties, so that the secondary structure and hydepathic properties of the polypeptide are not substantially altered by those skilled in the peptide chemistry arts! , Replace as expected.
  • the following groups of amino acids that are conservative substitutions for each other are generally known: (1) glycine, asparagine, gnoletamine, cystine, serine, threonine and tyrosine; (2) alanine, parin, leucine, isoleucine, (3) Glycine, Endolanin, Serine, Threonine and Methionine; (4) Leucine, Isoleucine and Parin; (5) Glutamine and Asparagine; (6) Glutamate and Aspartate; 7) Arginine, lysine and histidine; (8) Feruaranine, tryptophan and tyrosine.
  • the peptide of the present invention only needs to have the above-mentioned amino acid sequence to the extent that a desired action and effect can be obtained, and may further include an additional sequence.
  • a person skilled in the art can naturally understand how much sequence is allowed to be added to the N-terminal side and C-terminal side, respectively, in order to achieve a desired effect as an antigen peptide. Therefore, for example, an amino acid sequence useful for promoting immunization, an amino acid sequence that facilitates formulation, or a peptide as a fusion protein on the N-terminal side and / or C-terminal side of the peptide according to the present invention.
  • amino acid sequence that is convenient for the expression of amino acids or an amino acid sequence that is convenient for the production and purification of peptides may be added.
  • the present invention As long as the specificity of binding to the antibody is not lost, the peptide may be chemically modified, or it may be added with a polymer or sugar chain.
  • the HCV peptide according to the present invention is a gene recombination technique using a normal chemical synthesis method, an enzymatic decomposition method of a protein molecule, or a host transformed to express a base sequence encoding the target amino acid sequence. Etc. can be manufactured.
  • the target peptide When the target peptide is produced by a chemical synthesis method, it can be produced by a known and commonly used technique in ordinary peptide chemistry, for example, using a peptide synthesizer. Thus, it can be synthesized by a solid phase synthesis method.
  • the crude peptide thus obtained can be purified by purification methods commonly used in protein chemistry, such as salting-out, ultrafiltration, reverse phase chromatography, ion exchange chromatography, and affinity. It can be purified by chromatographic methods.
  • a desired peptide is produced by gene recombination technology
  • a synthetic or cloned DNA fragment encoding the target amino acid sequence is incorporated into an appropriate expression vector, and this expression vector is used.
  • the desired peptide can be obtained by transforming microorganisms or animal cells using, and culturing the resulting transformant.
  • expression vectors that can be used plasmids, virus vectors, and the like known in the art can be used.
  • Host cell transformation methods using expression vectors in this peptide production technique include methods known per se, such as the calcium chloride method, calcium phosphate coprecipitation method, DEAE dextran method, lipofectin method, The positioning method can be used and should be selected as appropriate based on the host cell used.
  • the obtained peptide can be purified from the cell extract or culture supernatant collected from the cultured medium by the purification method described above.
  • the peptide of the present invention is useful as a pharmaceutical composition for treating a patient having an HCV-related disease, that is, as a peptide vaccine.
  • a vaccine consisting of a hepatitis C virus-derived peptide is prepared by mixing the peptide produced as described above with a pharmaceutically acceptable adjuvant and Z or a carrier as appropriate.
  • Adjuvants include adjuvants that can enhance the immune response, such as Freund's incomplete adjuvant, aluminum hydroxide gel. Can be used.
  • the carrier for example, phosphate buffered saline (
  • PBS distilled water
  • physiological saline physiological saline
  • the peptide vaccine can be administered by a transdermal route such as oral or intravenous administration or subcutaneous administration, depending on the form of use.
  • a transdermal route such as oral or intravenous administration or subcutaneous administration, depending on the form of use.
  • the dosage form include tablets, granules, soft capsules, hard capsules, liquids, oils, and emulsifiers.
  • the dosage of such a pharmaceutical composition may vary depending on the symptoms of the patient to whom it is administered, but in general, 0.1 to 10 mg of peptide per day is preferable for adults.
  • the interval is preferably administered once every few days to several months. You can also use peptide vaccine and interferon together.
  • the hepatitis C virus-derived peptide to be administered is selected based on the peptide reactivity of anti-HCV antibodies present in the patient's blood or the presence of CTL. More preferably, the selection is based on the peptide reactivity of the anti-HCV antibody and the presence of CTL.
  • “peptide reactivity of anti-HCV antibody” means that any of the peptides to be administered is recognized by the anti-HCV antibody present in the blood of the patient. Higher therapeutic effects can be expected by examining the peptide reactivity of anti-HCV antibodies in each patient and administering only the appropriate peptides for each patient, so-called tailor-made administration.
  • the tailor-made administration induces secondary immunity promptly, resulting in a safer and better clinical effect (Mine et al., Clin Can. Res. 929-937, 2004).
  • HCV-related diseases can be confirmed by monitoring the blood antibody titers of subjects suffering from HCV-related diseases, that is, the blood concentration of antibodies reactive to peptides, by conventional methods such as ELISA. Can be done.
  • the amount of virus can be monitored by measuring the amount of HCV RNA by conventional methods such as RT-PCR.
  • the peptide of the present invention is also useful as a composition for diagnosing a patient having an HCV-related disease or predicting the progression of the disease.
  • the antibody titer in the blood of a subject can be determined using an antigen-antibody reaction well known in the art Can be measured.
  • the measurement can be performed as follows.
  • the antigen peptide is bound to a conventional ELISA plate such as 96well, and the plate is appropriately blocked to prevent nonspecific adsorption.
  • dilute the prepared blood serum of the test subject appropriately and cover each well of the plate and react for a predetermined time.
  • the plate After washing the plate to remove unbound components, add antibodies that can bind to human antibodies (eg, rabbit anti-human antibodies). If it is desired to detect IgG, ⁇ chain specific anti-HgG can be used. After reacting for a specified time, the plate is washed and detectably labeled antibody (eg anti-rabbit IgG) is added. Labeling can be performed by methods well known to those skilled in the art using enzymes, fluorescent dyes, chemiluminescent substances, piotin, radiation compounds and the like. After reacting the plate for a specified time, the label is detected by adding an appropriate substrate and measuring the decrease in substrate or increase in product, or by measuring fluorescence, luminescence, or radioactivity. In this way, the amount of antibody against a specific peptide in the serum of the subject can be measured. In addition, the progression of HCV-related disease can be predicted by monitoring the amount of antibody.
  • detectably labeled antibody eg anti-rabbit IgG
  • Labeling can be performed by methods well known to
  • xMAP technology is one of highly sensitive methods. This is a flowmetry measurement method developed by Luminex using a fluorescent microbead array system. Microbeads to which peptides are bound are brought into contact with serum, and then a fluorescently labeled secondary antibody is bound to the flow. Measure fluorescence intensity by measurement.
  • an immunochromatography method can be used.
  • a portion in which the antigen (or antibody) is linearly distributed is made on the test paper, and the complex of the antibody in the sample and the antigen labeled with the colored particles moves on the test paper.
  • Qualitative analysis is based on the presence or absence of colored lines that appear by intensive capture by antigens (or antibodies). Using this method, results can be obtained in a short time (within 20 to 30 minutes) with simple equipment.
  • the subjects were 10 HLA-A2-positive HCV2a-infected patients and 10 healthy HLA-A2-positive donors.
  • HCV2a patients were confirmed to be positive for anti-HCV antibodies by immunoassay.
  • HCV2a-derived peptides used in the following Examples. These peptides are 9 types of 9-mer or 10-mer synthetic peptides with HLA-A2 binding motif derived from HCV-genotype 2a protein.
  • the positive control included an influenza (Flu) virus-derived peptide (GILGFVFTL (SEQ ID NO: 10)), an EBV-derived peptide (GLCTLVAML (SEQ ID NO: 11)) having an HLA-A2 binding motif, and an HIV-derived peptide (SLYNTVATL ( SLYNTVATL ( SEQ ID NO: 12)) was used.
  • peptides were purchased from BIOSYNTHESIS (Lewisville, TX) or SynPep (Dublin, CA) and were more than 90% pure. The peptide was dissolved in dimethyl sulfoxide (DMSO) at a concentration of lOmgZml and stored at 120 ° C.
  • DMSO dimethyl sulfoxide
  • T2 and HEK293-A2 cell lines expressing HLA-A2, were maintained in RPMI-1640 medium supplemented with 10% FCS.
  • HEK293-A2 cells 0.5 gZ ml of hygromycin B (GibcoBRL, New York, NY, USA) was added to the culture.
  • HCV2a subgenome Plasmid pSGR- containing the conserved sequence of NS 3— NS 5 of JFH— 1 HI is described in Kato et al. (Gastroenterology. 2003; 125 (6): 1808-1817). The plasmid was linearized by Xbal digestion, and then Mangbean 'nuclease (New England
  • HEK293 A2 cells treated with trypsin were washed with PBS and added to ice-cold Cytomix buffer (v an den Hoff MJ, et al, Nucleic Acids Res. 1992; 20 (11): 2902) 0.5—lxl 0 7 / ml Resuspended at a concentration of Synthetic levulincon RNA (5 ⁇ g) is mixed with 400 ⁇ 1 cell suspension, transferred to an erect-portion cuvette (Bio-Rad, Hercules, CA), and Gene Pulser II instrument (Bio-Rad). A pulse was applied at 260V and 950 F using Next, the transferred cells were transferred to RPMI1640 medium containing 10% FCS and cultured in a culture flask. 18—24 hours after transfusion, G418 (0.5 mg / ml) (Nacalai
  • RNA—Bee® TEL—TEST, TX, USA
  • CDNA was synthesized from 5 ⁇ g of total RNA.
  • HCV cDNA consists of a set of oligonucleotide primers specific for HCV2a (forward primer, nucleotide position 7244— 7263: 5′— AGGAGGCCAGAT TACCAACC— 3 ′ (SEQ ID NO: 13), reverse primer, nucleotide position, 7376— 7395: 5 '-AAGGTCTTG ATGGCC AGTTG-3' (SEQ ID NO: 14) Detected by amplification.
  • PCR was performed with TaqDNA polymerase (Promega, WI, USA) for 30 cycles (95 ° C for 1 minute, 60 ° C for 1 minute, 72 ° C for 1 minute). PCR products were analyzed on 2 o / o agarose gels.
  • Medium is 45% RPMI—1640, 45% IV—V medium (QbcoBRL, NY, USA), 10% FCS, lOUU / ml interleukin 2 (IL—2), and 40 g / ml gentamicin strength.
  • RPMI—1640 45% RPMI—1640, 45% IV—V medium (QbcoBRL, NY, USA)
  • 10% FCS lOUU / ml interleukin 2 (IL—2)
  • IL—2 interleukin 2
  • 40 g / ml gentamicin strength Become. Every 3-4 days, half of the culture was removed and replaced with fresh medium containing the corresponding peptide at 20 / z gZml. On the 15th day, half of the cultured cells in the well were equally divided into four wells. Two wells were further stimulated with T2 cells pulsed with the corresponding peptide, and the other two wells were stimulated with T2 cells pulsed with the control HIV
  • PBMCs stimulated with the HCV2a peptide were cultured for about 2-3 weeks using irradiated HLA-A2-positive buffy coat cells as feeder cells to obtain a sufficient number of cells for analysis of cytotoxic activity.
  • PBMCs isolated from HLA-A2-positive healthy donors are seeded in culture plates at 4xl0 6- cell Zml in 10% FCS, PRMI1640 medium, incubated at 37 ° C for 60-90 minutes, and the cells are plated on the culture plates. Attached. The non-adherent cells were removed by gentle washing, and the attached monocyte-rich population was cultured in the following medium for 7 days.
  • the DCs are washed with PBS, 1. Resuspended in Oml medium and incubated with 3 gZml j8 2-microglobulin (Sigma) and 10 ⁇ gZml peptide for 2 hours at 37 ° C. I started. After irradiation (40 Gy), DC were further incubated with a non-adherent PBMC in a ratio of 1:20. On days 7, 14 and 21, the PBMC cultures were restimulated with a DC pulsed with peptide and irradiated at a ratio of 1:40. On day 28, a cytotoxic activity assay was performed.
  • HLA-A2-binding putative peptides derived from HCV2a were analyzed by BIMAS software (Bioinfomatics and Molecular Analysis Section, NIH). Peptide bond scores were calculated based on the predicted half-life of dissociation from HLA class I molecules (Bioinformatics and Molecular Analysis Section, Division of Computer Research & Technology, NIH). [0046] The ability of these peptides to induce PBMC force CTL in 10 HLA-A2-positive HCV2a-infected patients and 5 HLA-A2-positive healthy donors was examined. PBMC from patients and healthy donors were stimulated in vitro with each peptide for 2 weeks and then tested for reactivity to the corresponding peptide.
  • Table 9 shows the BIMAS binding scores for the nine types of peptides and their peptides—half-life of MHC dissociation in which CTL induction was observed.
  • Table 2 shows the results of CTL induction. The values in Table 2 represent IFN- ⁇ produced from effector PBMC in response to T2 cells pre-pulsed with the corresponding peptide. The IFN- ⁇ response to ⁇ 2 cells pre-pulsed with control HIV peptide was subtracted as background. Significant values (two-sided Student t test ⁇ ⁇ 0.05) are underlined. Pt; HCV2a infected patients, HD; healthy donors, ND; not measured.
  • the nine peptides of the present invention were able to induce peptide-specific CTLs from PBMC cultures of at least one patient.
  • the HCV2a432-441 peptide most effectively induced peptide-specific CTL, which generated peptide-specific CTL from 8 out of 9 HLA-A2-positive patients.
  • HCV2a716-724 and HCV2a 2251-2260 peptides induced peptide-specific CTL from 4 of 8 patients and 5 of 9 patients, respectively.
  • the ratio of positive responses was less than 50%.
  • these nine peptides were able to induce peptide-specific CTLs from healthy donor PBMCs.
  • HLA-A2-positive healthy donor PBMCs were stimulated in vitro with DC pulsed with HCV2a peptide and their cytotoxic activity was examined by 6 hr 51 Cr-release assay.
  • T2 cells pulsed with the corresponding HCV2a peptide or control HIV peptide were used.
  • a typical result is shown in Figure 2A.
  • In vitro stimulation of DC pulsed with HCV2a432—441, HCV2a716-72 4, or HCV2a2251—2260 peptide PBMCs in a safe donor could induce peptide-specific CTL.
  • the values shown are the average of three measurements, and statistical analysis was performed by Student's t test (* P ⁇ 0. 05).
  • CD8 + cells of PBMC stimulated with HCV2a peptide were purified and examined for cytotoxic activity against T2 cells norsted with the corresponding HCV2a peptide in the presence of various monoclonal antibodies.
  • Purified CD8 + T cells showed peptide-specific cytotoxic activity, which was blocked by the anti-HLA class I antibody, but not blocked by the anti-HLA class II antibody ( ( Figure 2B). These results indicate that peptide-specific CD8 + T cells show cytotoxic activity in an HLA class I-restricted manner.
  • HCV2aRNA HEK293-A2 cells expressing HCV2aNS3-NS5 were used as target cells, and the cytotoxic activity of CTL induced by the HCV2a2251-26-2 peptide derived from the NS5 protein of HCV2a was measured.
  • HCV2aNS3-NS5 mRNA in HEK293-A2 cells and parental HEK2 93-A2 cells transfected with HCV2aNS3-NS5 was examined by RT-PCR.
  • As a control j8-actin was used. PCR products were analyzed on 2% agarose gels. As a result, the presence of HCV2a subgenomic RNA was confirmed in stably transfected HEK293-A2 cells (Fig. 3A).
  • HCV2a peptide was naturally processed and presented on HCV2a-infected cells, and that HCV2a2 251-2260 peptide-specific CTL was able to lyse HCV2a-infected cells.
  • the HCV2a peptide of the present invention has immunogenicity, and thus is considered useful for specific immunotherapy of HLA-A2-positive HCV2a-infected patients.
  • the peptides of the present invention are useful for the treatment of HCV-related diseases and the diagnosis or prediction of progression of HCV-related diseases.

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Abstract

La présente invention concerne un peptide contenant une séquence d'acides aminés sélectionnée parmi les séquences suivantes : E1 284-293: VMLAAQMFIV ; E1 285-293: MLAAQMFIV ; E2 432-441: SLHTGFLASL ; E2 716-724: YIVRWEWVV ; NS2 888-897: VVFDITKWLL ; NS5 2251-2260: VVLDSLDPMV ; NS5 2850-2858 ; WLGNIIQYA ; et NS5 3012-3021: RLLLLGLLLL. Le peptide selon l'invention peut être administré à un sujet infecté par le VHC 2a. L'invention a également trait à une composition pharmaceutique destinée à traiter un patient atteint d'une maladie associée au VHC, ou à une composition destinée à prédire l'évolution d'une maladie associée au VHC, lesdites compositions contenant le peptide selon l'invention comme ingrédient actif.
PCT/JP2007/050966 2006-01-23 2007-01-23 Peptide antigène à restriction hla-a2 dérivés du virus de l'hépatite c 2a Ceased WO2007083806A1 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009069302A1 (fr) * 2007-11-28 2009-06-04 Oncotherapy Science, Inc. Peptides épitopes de stat3
WO2009150822A1 (fr) * 2008-06-10 2009-12-17 Oncotherapy Science, Inc. Peptides épitopes dérivés de mybl2 et vaccins les contenant

Citations (1)

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Publication number Priority date Publication date Assignee Title
WO2005028503A1 (fr) * 2003-09-22 2005-03-31 Green Peptide Co., Ltd. Peptide provenant du virus de l'hepatite c

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Publication number Priority date Publication date Assignee Title
WO2005028503A1 (fr) * 2003-09-22 2005-03-31 Green Peptide Co., Ltd. Peptide provenant du virus de l'hepatite c

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WO2009069302A1 (fr) * 2007-11-28 2009-06-04 Oncotherapy Science, Inc. Peptides épitopes de stat3
WO2009150822A1 (fr) * 2008-06-10 2009-12-17 Oncotherapy Science, Inc. Peptides épitopes dérivés de mybl2 et vaccins les contenant
RU2496787C2 (ru) * 2008-06-10 2013-10-27 Онкотерапи Сайенс, Инк. Пептиды эпитопов mybl2 и содержащие их вакцины

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