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WO2009148229A2 - Peptide immunogène et composition contenant le peptide destinée à prévenir ou à traiter des maladies liées au vph - Google Patents

Peptide immunogène et composition contenant le peptide destinée à prévenir ou à traiter des maladies liées au vph Download PDF

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Publication number
WO2009148229A2
WO2009148229A2 PCT/KR2009/002713 KR2009002713W WO2009148229A2 WO 2009148229 A2 WO2009148229 A2 WO 2009148229A2 KR 2009002713 W KR2009002713 W KR 2009002713W WO 2009148229 A2 WO2009148229 A2 WO 2009148229A2
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Prior art keywords
peptide
seq
peptides
hla
hpv
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WO2009148229A3 (fr
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이경률
임종백
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Biocore Co Ltd
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Biocore Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • 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
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
    • 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
    • A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/20011—Papillomaviridae
    • C12N2710/20022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • the present invention relates to an immunological peptide and a composition for preventing or treating HPV-related diseases comprising the peptide, and more particularly, to a composition for preventing or treating HPV-related diseases comprising an E7 immunopeptide derived from an E7 protein of HPV type 16 and the peptide. It is about.
  • Cervical cancer is the second most common malignant tumor in women, with more than 350,000 people dying from cervical cancer each year, and the number continues to increase (zur Hausen H .. Nat Rev). Cancer 2002. 2 (5): 342-50).
  • the main cause of cervical cancer is known as human papilomavirus (HPV), and it has been found that 40 out of 100 HPV infections cause female genital infections (de V Amsterdam EM, Fauquet C, Broker TR, Bernard HU and Kir). Hausen H. Classification of papillomaviruses. Virology. 2004. 324: 17-27).
  • HPV type 16 and HPV type 18 are most commonly found in cervical cancer patients and are known to be important in tumor development. Among them, HPV type 16 is detected in most cervical cancer patients. (Munoz N, et al. N. Engl. J. Med. 2003. 348: 518-527).
  • E6 and E7 are potent oncoproteins that inhibit the actions of tumor suppressors p53 and RB. It has been shown to promote the progression and continued growth of cancer cells (Werness BA, et al. Science. 1990. 248: 76-79; Dyson N, et al. Science. 1989. 243: 934-937). E7 inhibits the function of RB, a tumor suppressor (Kiyono T, et al. Nature. 1998. 396: 84-88), promotes the activities of the S-phase genes cyclin A and cyclin E (Zerank K, et al. J.).
  • HPV type 16 E7 epitope is proposed to induce the production of cytotoxic T lymphocytes that can be treated in cervical cancer patients with HLA-A * 0201 type.
  • the present invention has been made by the above necessity, an object of the present invention to provide a peptide for the prevention or treatment of HPV-related diseases.
  • the present invention to achieve the above object
  • Peptides containing the LEDLLMGTLGIVCPI amino acid sequence of SEQ ID NO: 13 or the GTLGIVCPICSQKP amino acid sequence of SEQ ID NO: 14 are provided.
  • the peptides of the present invention are not only peptides described in SEQ ID NO: 13 or SEQ ID NO: 14, but also one or more deletions, substitutions, and inversions of these peptides. It also includes mutant peptides that can achieve the object of the present invention by causing mutations, such as translocation.
  • the present invention also provides a nucleic acid encoding the peptide of SEQ ID NO: 13 or the peptide of SEQ ID NO: 14 or the above mutant peptides of the present invention, respectively.
  • the nucleic acid sequence is SEQ ID NO: 18 for the peptide of SEQ ID NO: 13 (ttg gaa gac ctg tta atg ggc aca cta gga att gtg tgc ccc atc) and SEQ ID NO: 19 for the peptide of SEQ ID NO: 14 ggc aca cta gga att gtg tgc cc atc tgt tct cag aaa cca) It is not limited to this.
  • the present invention also provides a composition for preventing or treating a disease associated with HPV infection, comprising (a) the peptide of the present invention and (b) a pharmaceutically acceptable carrier.
  • the HPV infection-related disease is bowenoid papulosis, anal dysplasia, respiratory or conjunctival papilloma, cervical dysplasia, cervical cancer, vulval cancer, Or prostate cancer, but is not limited thereto.
  • the present invention also provides a microparticle, liposome, or immune stimulating complex (ISCOM) comprising the peptide of the present invention as an active ingredient.
  • ISCOM immune stimulating complex
  • the present invention also provides a microparticle, liposome, or immune stimulating complex (ISCOM) comprising the nucleic acid of the present invention as an active ingredient.
  • ISCOM immune stimulating complex
  • the composition of the present invention is preferably HLA-A * 3303 type patient specific, but is not limited thereto.
  • the peptides or nucleic acids of the invention may optionally be formulated in microparticles, liposomes, or immune stimulating complexes (ISCOM) or other vehicles suitable for feeding the subject peptide to the subject.
  • microparticles it preferably has a polymer matrix which is a copolymer such as poly-lactic-co-glycolic acid (PLGA).
  • an immune response e.g., a cellular immune response comprising an MHC class I-mediated or class II-mediated immune response
  • an immune response is a mammal having an MHC molecule to which an immunogenic peptide binds, ie humans, apes, dogs, cats, and animals. It can be triggered by administering immunogenic peptides to cows, mice, and rats.
  • the peptide of the present invention may be administered as a microparticle, liposome, or ISCOM, or as part of a solution.
  • Another method of administering a peptide of the invention is to use an expression vector comprising a nucleic acid sequence encoding the peptide of the invention.
  • the nucleic acid sequence may optionally encode a signal sequence linked to the above peptide of the present invention.
  • the nucleic acid encodes a signal sequence, preferably it is a signal sequence Met Ala Ile Ser Gly Val from HLA-DR.alpha. Pro Val Leu Gly Phe Phe Ile Ala Val Leu Met Ser Ala Gln Glu Ser Trp Ala (SEQ ID NO: 20).
  • the sequence of the present invention may have the sequence of SEQ ID NO: 13 or 14, for example.
  • the nucleic acid does not contain the viral gene causing the nucleic acid infection and does not encode the intact E7 protein.
  • Nucleic acids of the invention can be included in the plasmid and optionally provided in microparticles comprising a polymer matrix.
  • the polymer matrix consists essentially of a copolymer of PLGA.
  • the microparticles have a diameter of, for example, 0.02 to 20 microns or less than about 11 microns.
  • the plurality of microparticles has a diameter of 0.02 to 20 microns or less than about 11 microns.
  • Cells containing the plasmid of the present invention are also within the scope of the present invention.
  • the cell can be, for example, a B cell or other antigen presenting cell (APC).
  • the cells are cultured or maintained under conditions that allow peptide expression from the plasmids encoding them.
  • Nucleic acids and plasmids of the present invention are useful in methods of inducing an immune response in mammals, eg, humans, by administering the aforementioned plasmids to the mammal, eg, as "naked DNA.”
  • the mammal may have a risk or disease of HPV infection, cervical dysplasia, and / or cervical cancer.
  • Nucleic acids and plasmids of the invention can also be inserted into microparticles, liposomes, ISCOMS, or other suitable means of transport as described above.
  • Peptides disclosed herein and nucleic acids encoding the peptides can be used to elicit an immune response against HPV E7 protein.
  • the peptide of the present invention may be linked to a transporting sequence that carries the peptide to intracellular organs.
  • the transport sequence is an amino acid sequence that functions to regulate intracellular transport (directing movement from organelles to organelles or to the cell surface) of the peptide to which it is attached.
  • Such transport sequences can transport the polypeptides to ER, lysosomes or endosomes and signal peptides (N-terminal sequences that direct the protein to the ER during translation), for example ER retention sequences such as KDEL, and KFERQ, Or a lysosomal target sequence such as QREFK.
  • Short amino acid sequences can act as signals to target proteins to specific intracellular organs. For example, a hydrophobic signal peptide is found at the amino terminus of the protein towards ER.
  • Such a transport sequence may be the HLA-DR.alpha leader sequence Met Ala Ile Ser Gly Val Pro Val Leu Gly Phe Ile Ala Val Leu Met Ser Ala Gln Glu Ser Trp Ala (SEQ ID NO: 20). If that portion is sufficient to transport the polypeptide of the invention to the ER, it may comprise only a portion of the above specified 25 residue sequences (eg at least 10 amino acid residues) of the signal peptide.
  • Standard techniques can be used to construct DNA encoding the peptides of the invention (see eg the technique described in WO 94/04171).
  • the construct can be located on additional sequences that increase expression in human cells, such as appropriate promoters, RNA stable 5 'and 3' sequences of coding sequences, introns (5 'or 3' in the sequence being encoded).
  • poly (A) addition sites, as well as their constructs may include replication origin and selection markers that allow for selection and replication for prokaryotic and / or eukaryotic hosts.
  • the plasmid of the present invention may have a kanamycin resistance gene (519-1313 site), an SV40 early promoter (131-484 site), and a thymidine kinase (TK) polyadenylation site (1314-1758 site).
  • Kanamycin resistance genes and related regulatory sequences are for selection purposes only and can be removed from the plasmid if selection is not required or desired.
  • the peptides and nucleic acids of the invention can be used as prophylactic or therapeutic vaccines in subjects known to be infected by HPV, suspected of being infected by HPV, or susceptible to infection by HPV. Other suitable subjects include those who exhibit or are likely to develop symptoms of HPV-related diseases.
  • the peptides and vaccines of the present invention are useful for treating diseases associated with infection of HPV strain 16, such as bowenoid papulosis, anal dysplasia, respiratory or conjunctival papillomas, cervical dysplasia, cervical cancer, and vulvar cancer. vulval cancer, or as a vaccine to treat or prevent prostate cancer.
  • Peptides or nucleic acids of the invention can be administered alone or in combination with other therapies known in the art, such as chemotherapeutic agents, radiation and surgery to treat HPV infection or diseases associated with HPV infection.
  • Peptides and nucleic acids of the invention can also be administered in combination with other therapies designed to enhance an immune response, for example adjuvant or cytokines (or nucleic acids encoding cytokines), as is well known in the art. .
  • Peptides or nucleic acids of the invention can be used in the manufacture of a medicament for the prevention or treatment of HPV infection or a disease associated with HPV infection.
  • the delivery systems of the present invention can be used to deliver a peptide or DNA construct that expresses the peptide to appropriate cells that are intended to promote an immune response against HPV.
  • Peptides of the invention or nucleic acids encoding the peptides are standard methods, for example Donnelly et al., J. Imm. Methods 176: 145, 1994, and Vitiello et al., J. Clin. Invest. 95: 341, may be administered using a method such as that described in 1995.
  • Peptides or nucleic acids of the invention can be injected in a form known in the art to a subject, such as intramuscular injection, intravenous injection, arterial injection, intradermal injection, intraperitoneal injection, intranasal, intravaginal, enema, subcutaneous or they For example, it may be administered to the gastrointestinal tract, mucosa or respiratory tract by inhalation of a powder or solution containing microparticles. Administration can be local (eg cervical or other infectious site) or systemic.
  • the peptides of the present invention or nucleic acids encoding the peptides can be carried in pharmaceutically acceptable carriers such as colloidal suspensions, powders, saline, lipids, liposomes, microspheres, or nanospheres. They may be complexed with, associated with, or naked with a vehicle and may be lipids, liposomes, microparticles, gold, nanoparticles, polymers, catalyzers, polysaccharides, polyamino acids, dendrimers, saponins, adsorption enhancing substances or fatty acids. The delivery can be carried out using a delivery system known in the art.
  • a dose of about 0.1 to 100 micromoles of peptide or about 1 to 200 micrograms of DNA is administered per kg of body weight once.
  • a dose for a given patient may be based on a number of factors including the patient's height, body surface area, age, the specific compound administered, sex, time and route of administration, general health and other agents administered simultaneously. Depends on Determination of the appropriate dosage is skillful within the capabilities of pharmacists of ordinary skill.
  • APCs antigen presenting cells
  • dendritic cells dendritic cells
  • peripheral blood mononuclear cells or bone marrow cells
  • APCs antigen presenting cells
  • bone marrow cells can be obtained from a patient or a suitable donor and activated in vitro with the present immune composition and then administered to the patient. have.
  • Microparticles including those described in US Pat. No. 5,783,567, can be used as vehicles for transporting macromolecules such as DNA or peptides into cells. They contain macromolecules enclosed within the shell of the polymer or entered into the polymer matrix. Microparticles act to maintain the properties of macromolecules, for example to keep the DNA intact. Microparticles can also be used for pulsed delivery of macromolecules and for delivery to specific locations or to specific cells or target cell populations.
  • the polymer matrix may be a biodegradable copolymer such as poly-lactic-co-glycolic acid, starch, gelatin or chitin.
  • Microparticles can be used in particular to maximize the transport of DNA molecules into the phagocytes of the subject. Or the microparticles can be injected or implanted into tissue.
  • Peptides of the invention can be administered to a subject via lipids, dendrimers or liposomes well known in the art.
  • liposomes carrying an immunopeptide or nucleic acids encoding the peptide are known to cause a CTL response in vivo (Reddy et al., J. Immunol. 148: 1585, 1992; Collins et al., J. Immunol. 148: 3336-3341, 1992; Fries et al., Proc. Natl. Acad. Sci. USA 89: 358, 1992; Nabel et al., Proc. Nat. Acad. Sci. (USA) 89: 5157, 1992).
  • Peptides and nucleic acids of the invention can be administered using Immune Stimulating Complexes (ISCOMS), a 30-40 nm sized (-) occupied cage-like structure produced by mixing saponin alone or cholesterol and Quil A (saponin) simultaneously. have. Peptides and nucleic acids of the invention can be administered simultaneously or separately from ISCOMS.
  • ISCOMS Immune Stimulating Complexes
  • the ability of the peptides and nucleic acids of the invention to elicit an immune response can be analyzed using methods for measuring immune responses well known in the art.
  • the generation of cytotoxic T cells can be expressed using MHC tetramers, measuring intracellular cytokine expression or in a standard 51 Cr release assay.
  • Standard assays such as ELISA or ELISPOT can also be used to determine cytokine profiles that contribute to T cell activation.
  • T cell proliferation can also be measured using other assays known in the art and assays such as the 3 H-thymidine uptake.
  • B cell responses can be measured using an assay such as ELISA.
  • Cervical cancer is the second most common malignant tumor in women and its risk is well known.
  • Human papillomavirus (HPV) is known to be the major cause of cervical cancer, among which HPV type 16 has been found to be the most detected in cervical cancer patients.
  • HPV types 16 E6 and E7 are viral oncoproteins that have been shown to cause HPV-infected cervical cells to progress into cancer cells and promote the continued growth of cancer cells.
  • HPV type 16 E7 inhibits the function of Rb, a tumor suppressor, inhibits apoptosis of infected cells and induces the transition to tumors.
  • HPV type 16 E7 is a potent target antigen in the development of immune cell therapy using cytotoxic T lymphocytes for the treatment of cervical cancer. It is being studied as a target antigen.
  • E7 candidate peptides as a result of flow cytometry using HPV type 16 E7 candidate peptides synthesized to develop specific HPV type 16 E7 epitopes that induce the production of HLA-A * 0201 type cytotoxic T lymphocytes was found that the induction of the E7 79-93 (LEDLLMGTLGIVCPI) and E7 85-98 (GTLGIVCPICSQKP) epitopeuyi higher cell differentiation and activity of cytotoxic T cells, and E7 79-93 through cytotoxicity (cytotoxicity) experimental ( The LEDLLMGTLGIVCPI) and E7 85-98 (GTLGIVCPICSQKP) epitopes showed that the E7 82-90 (LLMGTLGIV) and E7 86-93 (TLGIVCPI) peptides, which were well identified in previous studies, caused the same or superior cervical cancer cell death.
  • a PBMC in order to find a peptide having a higher anticancer effect than the known E7 epitope, a PBMC was supplied from a normal HLA-A * 0201 type blood donor and treated with a synthesized E7 peptide, followed by incubation for 1 week or 2 weeks, and then IFN- ⁇ expression was measured to select candidate peptides to induce differentiation into CD8 + T cells, and the actual anti-cancer effect was confirmed using 51 Cr release assay, a cytotoxicity test.
  • E7 79-93 LEDLLMGTLGIVCPI
  • E7 85- 98 GTLGIVCPICSQKP
  • E7 79-93 CMVpp65 in the experimental group treated with the peptide 495-503 significantly higher cell killing effect than the positive control group treated with (A02, NLVPMVATV) peptide ( Figures 3 and 4).
  • the fluid in the cell measurement and cell vaccines and adoptive immune cells, treatment of the HPV type 16 E7 E7 79- 93 (LEDLLMGTLGIVCPI) and E7 85-98 (GTLGIVCPICSQKP) peptide is HLA-A * 0201-type cervical cancer through toxicity test It was found that it was an effective epitope for the production of the necessary cytotoxic T lymphocytes.
  • HPV type 16 E7 E7 79-93 (LEDLLMGTLGIVCPI) and E7 85-98 (GTLGIVCPICSQKP) peptide is HLA-A * 0201-type vaccine development of cervical cancer and cytotoxic T required for adoptive immune therapy cell The epitope was found to be effective for lymphocyte production.
  • Figures 1 and 2 measure the degree of IFN- ⁇ expression in HLA-A * 0201 type specific CD8 + T cells induced by HPV type 16 E7 candidate epitopes using fluid cytometry.
  • FIG. 1 shows HLA-A * 0201 type specific CD8 + T cells induced after stimulation of peripheral blood monocytes extracted from HLA-A * 0201/2401 type patients with HPV type 16 E7 candidate peptides and negative and positive control peptides. It is a measure of the degree of IFN- ⁇ expression.
  • IL-2 negative control
  • A02 CMVpp65 495-493
  • HLA-A * 0201, NLVPMVATV positive control
  • A24 CMVpp65 328-336
  • HLA-A * 2402, QYDPVAALF positive control
  • A33 CMVpp65 91-100 (HLA-A * 3303, SVNVHNPTGR, Negative Control)
  • FIG. 2 shows the intracellular HLA-A * 0203 type specific CD8 + T cells induced after stimulation of peripheral blood monocytes extracted from HLA-A * 0201/3303 type patients with HPV type 16 E7 candidate peptides and negative and positive control peptides. It is a measure of the degree of IFN- ⁇ expression.
  • IL-2 negative control
  • A02 CMVpp65 495-493
  • A24 CMVpp65 328-336
  • HLA-A * 2402, QYDPVAALF negative control
  • A33 CMVpp65 91-100 (HLA-A * 3303, SVNVHNPTGR, positive control).
  • HLA-A * 0201 type cervical cancer cells measured after stimulation of peripheral blood monocytes extracted from HLA-A * 0201/3303 type patients with HPV type 16 E7 candidate peptides and negative and positive control peptides. being.
  • IL-2 negative control
  • A02 positive control
  • CMVpp65 495-493 HLA-A * 0201, NLVPMVATV, positive control
  • HLA-A * 0201 type cervical cancer cells measured after stimulation of peripheral blood monocytes extracted from HLA-A * 0201/2402 type patients with HPV type 16 E7 candidate peptides and negative and positive control peptides. being.
  • IL-2 negative control
  • A02 positive control
  • CMVpp65 495-493 HLA-A * 0201, NLVPMVATV, positive control.
  • HPV type 16 E7 protein The entire amino acid sequence of HPV type 16 E7 protein was synthesized by using computer algorithms to overlap each of 15mer in size, and then purified using a high performance liquid chromatography (HPLC) instrument. A total of 14 peptides were synthesized by finally selecting more than 95% of the peptides (Table 1). Peptide synthesized in this way was dissolved in 1% DMSO PBS and stored frozen at -20 °C and thawed at room temperature immediately before use.
  • HPLC high performance liquid chromatography
  • Table 1 shows the synthesized HPV type 16 E7 peptide.
  • PBMCs Peripheral blood mononuclear cells
  • Table 2 shows HLA-A * 0201 type blood donors.
  • Example 3 Generation of autologous dendritic cells from PBMC
  • PBMC obtained by centrifugation was placed in a T75 flask containing RPMI culture (10% FBS, 5% antibiotics) and incubated at 37 ° C. for 2 hours. Thereafter, the suspended cells were removed, and interleukin-4 (IL-4, 1000 U / ml) and granulocyte-macrophage stimulating factor (GM-CSF, 800 U / ml) were added to the monocytes fixed in the flask. IL-4 (1000U / ml) and GM-CSF (1600U / ml) were added at 2 and 4 days after incubation. RPMI culture medium was changed from time to time as the cells grow. At 5 days after the start of the culture, tumor necrosis factor- ⁇ (TNF- ⁇ , 1000u / ml) was added.
  • TNF- ⁇ tumor necrosis factor- ⁇
  • PBMCs of cryofreezing HLA-A * 0201 type blood donors were thawed and placed in the appropriate number of cells in a 24 well culture vessel filled with 2 ml RPMI culture medium per well.
  • the peptide (10 ⁇ g / ml) and Interleukin-2 (IL-2, 100 U / ml) to be tested were added to each well.
  • IL-2 (1000U / ml) was added at intervals of two days during the incubation period, and the culture medium was replaced.
  • the dendritic cells at least 10/10 the number of cytotoxic T cells
  • peptides (20 ⁇ g / ml) generated by the above-mentioned method were added.
  • dendritic cells and cytotoxic T cells to which the peptide was added were incubated at 37 ° C. for 1 hour.
  • phytohemagglutinin PHA, 0.25 ⁇ g / ml
  • BFA brefeldin A
  • cells were collected from each well and washed with PBS (phosphate-buffered saline). After washing, put PBS containing 1 mM concentration of EDTA and incubated for 10 minutes at 37 °C.
  • the cells were washed twice with PBS containing 5% FBS, and a fluorescently labeled antibody, perdinin-chlorophyll-protien (PerCP) -conjugated mouse anti-human CD3 + antibody and phycoerythrin (PE) -conjugated mouse anti-human CD8 + antibodies were put in a ratio of 1: 100, respectively, and then incubated in a dark state at 4 ° C. for 15 minutes. After incubation, the lysing solution and permeabilization solution were treated, and fluorescein isothiocyanate (FITC) -conjugated mouse anti-human IFN- ⁇ antibody was added and incubated in dark at 4 ° C for 30 minutes. Thereafter, the cells were washed twice with PBS containing 5% FBS, fixed with 1% formaldehyde, and analyzed by FACS (fluorescence activated cell sorting).
  • PerCP perdinin-chlorophyll-protien
  • PE phycoerythrin
  • Cytotoxicity experiments were performed using a 51 Cr release assay using cervical cancer cell lines as target cells. Briefly, 0.1 mCi Na 2 51 CrO 4 was added to HLA-A * 3303 type cervical cancer cell line cultured in RPMI culture medium and incubated at 37 ° C. for 45 minutes. After washing twice with PBS, an appropriate number of target cells were put in various ratios to cytotoxic T cells and incubated at 37 ° C. for 5 hours. After incubation, centrifuge at 1500 rpm for 5 minutes and transfer 100 ⁇ l to the prepared ⁇ -counter tube. Measure and analyze using ⁇ -ray counter. The extent of cytotoxic T cell specific target cell death is calculated using the formula [(experimental cpm-spontaneous cpm) / (maximum cpm-spontaneous cpm)] ⁇ 100.
  • each peptide was treated with PBMCs of blood donor for 1 week. Thereafter, the control groups treated with the cultured IL-2 (negative control) and CMVpp65 495-503 (NLVPMVATV, HLA-A * 0201, positive control) were compared. As shown in Figures 1 and 2, most of the 14 candidate peptides induced the production of IFN- ⁇ higher than the control group treated with IL-2 only, and peptides showing higher levels of IFN- ⁇ production than the negative control.
  • E7 79-93 LEDLLMGTLGIVCPI
  • GTLGIVCPICSQKP E7 85-98
  • HLA-A * 0201 type cervical cancer cell line acts on apoptosis, it was analyzed using 51 Cr release assay. Giving process in PBMC of the blood donor to IL-2, CMVpp65 495-503 (NLVPMVATV , HLA-A * 0201, positive control), E7 79-93 (LEDLLMGTLGIVCPI) and E7 85-98 (GTLGIVCPICSQKP) peptides each culture after 1 week Each peptide was treated with dendritic cells of the same patient once again, followed by incubation for one week, and then mixed with 51 Cr-labeled HLA-A * 0201 type cervical cancer cell line and incubated for 5 hours using a gamma radiation meter. Analyzed.
  • IL-2 negative control
  • CMVpp65 495-503 NLVPMVATV, HLA-A * 0201, positive control
  • SEQ ID NO: 17 E7 82-90
  • E7 86-93 TLGIVCPI, positive control
  • E7 79-93 LEDLLMGTLGIVCPI
  • GTLGIVCPICSQKP E7 85-98

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Abstract

La présente invention concerne un peptide immunogène et une composition contenant le peptide destinée à prévenir ou à traiter des maladies liées au VPH, et plus particulièrement, un peptide immunogène E7 dérivé d'une protéine E7 de type 16 du VPH, ainsi qu'une composition contenant le peptide destinée à prévenir ou à traiter des maladies liées au VPH.
PCT/KR2009/002713 2008-06-02 2009-05-22 Peptide immunogène et composition contenant le peptide destinée à prévenir ou à traiter des maladies liées au vph Ceased WO2009148229A2 (fr)

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KR1020080051833A KR20090125629A (ko) 2008-06-02 2008-06-02 면역성 펩타이드 및 그 펩타이드를 포함하는 ｈｐｖ 관련질환 예방 또는 치료용 조성물
KR10-2008-0051833 2008-06-02

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017220463A1 (fr) 2016-06-20 2017-12-28 Isa Pharmaceuticals B.V. Formulation d'un vaccin peptidique.
WO2018085751A1 (fr) * 2016-11-07 2018-05-11 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Développement d'épitopes agonistes du papillomavirus humain
US11072660B2 (en) 2016-10-03 2021-07-27 Juno Therapeutics, Inc. HPV-specific binding molecules
US11471489B2 (en) 2018-04-05 2022-10-18 Juno Therapeutics, Inc. T cell receptors and engineered cells expressing same
US11952408B2 (en) 2017-10-03 2024-04-09 Juno Therapeutics, Inc. HPV-specific binding molecules
WO2024223299A2 (fr) 2023-04-26 2024-10-31 Isa Pharmaceuticals B.V. Procédés de traitement du cancer par administration de compositions immunogènes et d'un inhibiteur de pd-1

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JP2006512300A (ja) * 2002-10-03 2006-04-13 エピミューン インコーポレイテッド Ｈｌａ結合ペプチド及びその使用
KR20090103571A (ko) * 2008-03-28 2009-10-01 바이오코아 주식회사 면역성 펩타이드 및 그 펩타이드를 포함하는 ｈｐｖ 관련질환 예방 또는 치료용 조성물

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017220463A1 (fr) 2016-06-20 2017-12-28 Isa Pharmaceuticals B.V. Formulation d'un vaccin peptidique.
EP3552623A1 (fr) 2016-06-20 2019-10-16 ISA Pharmaceuticals B.V Formulation d'un vaccin peptidique
US11072660B2 (en) 2016-10-03 2021-07-27 Juno Therapeutics, Inc. HPV-specific binding molecules
WO2018085751A1 (fr) * 2016-11-07 2018-05-11 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Développement d'épitopes agonistes du papillomavirus humain
US11311613B2 (en) 2016-11-07 2022-04-26 The United States of Americans represented by the Secretary, Department of Health and Human Services Development of agonist epitopes of the human papillomavirus
US11952408B2 (en) 2017-10-03 2024-04-09 Juno Therapeutics, Inc. HPV-specific binding molecules
US11471489B2 (en) 2018-04-05 2022-10-18 Juno Therapeutics, Inc. T cell receptors and engineered cells expressing same
WO2024223299A2 (fr) 2023-04-26 2024-10-31 Isa Pharmaceuticals B.V. Procédés de traitement du cancer par administration de compositions immunogènes et d'un inhibiteur de pd-1

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