WO2002026252A1 - Vaccin contre le virus de la grippe et son procede de preparation - Google Patents

Vaccin contre le virus de la grippe et son procede de preparation Download PDF

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Publication number
WO2002026252A1
WO2002026252A1 PCT/CN2001/001218 CN0101218W WO0226252A1 WO 2002026252 A1 WO2002026252 A1 WO 2002026252A1 CN 0101218 W CN0101218 W CN 0101218W WO 0226252 A1 WO0226252 A1 WO 0226252A1
Authority
WO
WIPO (PCT)
Prior art keywords
epitope
influenza virus
polypeptide
vaccine
influenza
Prior art date
Application number
PCT/CN2001/001218
Other languages
English (en)
Chinese (zh)
Inventor
Yinghua Chen
Yun Lu
Jian Ding
Original Assignee
Tsinghua University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from CNB001212931A external-priority patent/CN1163271C/zh
Priority claimed from CN 00121290 external-priority patent/CN1338307A/zh
Application filed by Tsinghua University filed Critical Tsinghua University
Priority to AU2002223398A priority Critical patent/AU2002223398A1/en
Publication of WO2002026252A1 publication Critical patent/WO2002026252A1/fr

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Classifications

    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • 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
    • A61K39/12—Viral antigens
    • A61K39/145—Orthomyxoviridae, e.g. influenza virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
    • A61K2039/6031—Proteins
    • A61K2039/6081—Albumin; Keyhole limpet haemocyanin [KLH]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/16011—Orthomyxoviridae
    • C12N2760/16022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/16011—Orthomyxoviridae
    • C12N2760/16034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • the present invention relates to a vaccine prepared by bioengineering technology and a preparation method thereof, in particular to an influenza vaccine and a preparation method thereof.
  • Influenza is a common and frequent viral infection that poses a great threat to humans, especially the elderly, children and the weaker.
  • the existing influenza vaccines and their design and preparation technologies are as follows:
  • Inactivated influenza virus vaccine which is currently widely used in adults, and is made from pathogens that have been inactivated by the influenza virus. However, the vaccine strain needs to be replaced in time with the antigenic variation of the influenza strain. Otherwise, the immune effect will not be guaranteed or even invalid. Due to the high variability of the influenza virus, it is difficult for this vaccine to ensure its preventive effect.
  • a live attenuated influenza virus vaccine The vaccine was used in large quantities in humans in I960. It was also made using pathogens that were attenuated by influenza viruses. For the same reasons as in the first case, the vaccine strain needs to be replaced in time with the antigenic variation of the influenza strain. At present, screening experiments for new vaccine strains are underway at home and abroad.
  • Influenza virus subunit vaccine This vaccine is currently used by children abroad. It is the use of genetically recombined virus surface antigens to induce immune protection. The virus surface antigens also need to be replaced in time with the antigenic variation of influenza strains, otherwise the immune effect will not be guaranteed or even invalid.
  • influenza vaccines currently in use are generally characterized by their inability to deal with the antigenic variation of influenza virus strains.
  • influenza viruses are highly variable and antigenic mutations are constantly occurring.
  • influenza virus membrane protein HA for example: HA92-105, KAYSNCYPYDVPDY; HA127-133, WTGVAQD; HA183-195, HHPITDSDQTRLY; HA317-341, ATGLRNIPSIQSRGLFGAIAGFIEG
  • HA92-105, KAYSNCYPYDVPDY HA127-133, WTGVAQD
  • HA183-195 HHPITDSDQTRLY
  • HA317-341 ATGLRNIPSIQSRGLFGAIAGFIEG
  • multi-epitope epitope polypeptides can induce high titers , Predefined, multi-neutralizing epitope-specific neutralizing antibodies.
  • the object of the present invention is to provide an influenza virus vaccine capable of coping with mutations of the influenza virus.
  • influenza virus vaccine which includes at least one neutralization table containing influenza virus cell membrane protein HA or M2 coupled to a carrier protein or carrier polypeptide to form a conjugate. Polypeptides or epitopes of variants and T cell epitopes of influenza virus.
  • the neutralizing epitope or variant epitope in the polypeptide and the T-cell epitope of the influenza virus are a single epitope repeated at least once or a multiple epitope repeated at least once.
  • the neutralizing epitope and variant epitope are selected from one or more of the following members:
  • the T-cell epitope of the influenza virus is selected from one or more of the following members:
  • the vaccine also contains acceptable pharmaceutical adjuvants.
  • Another object of the present invention is to provide a method for producing the above-mentioned influenza virus vaccine. This is achieved in this way.
  • a method for preparing an influenza virus vaccine basically includes the following steps:
  • the artificially synthesized polypeptide containing the neutralizing epitope or variant epitope of the antigenic region on the influenza virus membrane protein HA or M2 and the T cell epitope of the influenza virus is selected from:
  • the neutralizing epitope or variant epitope of the influenza virus membrane protein protein HA or M2 contained in the polypeptide and the T-cell epitope of the influenza virus are repeated at least once.
  • At least one repeated single-epitope or at least one repeated multi-epitope polypeptide are selected from one or more of the following members:
  • the vaccine of the present invention cleverly utilizes the neutralizing epitopes on the influenza virus membrane proteins HA and M2 and the influenza virus T cell epitopes, which can induce strong human body to produce a longer-lasting immune protection characteristic.
  • the theory of the epitope vaccine proposed by the inventors of the invention is that the epitope vaccine has a strong anti-mutation ability and can induce high titer, predefined, multiple neutralizing epitope-specific neutralizing antibodies (Immunology Today, 20 : 588-589, December 1999; Immunobiology, 201: 323-331, December 1999; Scand. J. Immunology. 2000, 51: 497-501; Int. Arch.
  • influenza vaccine of the present invention is a multiple vaccine that can stimulate the human body to produce a variety of neutralizing antibodies and cytotoxic T cells (CTL) to the influenza virus.
  • CTL cytotoxic T cells
  • the influenza virus One or more epitopes are mutated, and antibodies to the mutated influenza virus that correspond to other unmutated epitopes are still present in the human being injected with the vaccine.
  • the vaccine of the present invention has the following advantages: 1.
  • the vaccine is a highly efficient multiple vaccine that can stimulate the production of a variety of neutralizing antibodies and CTLs, and can effectively deal with the mutation of the virus, thereby overcoming the insufficient or even ineffective effects of existing influenza vaccines. Disadvantages.
  • the active ingredient of the vaccine is a neutralizing epitope or a variant epitope on the influenza virus membrane protein HA or M2 and a T cell epitope of the influenza virus, which are coupled to a carrier protein or carrier polypeptide to form a conjugate These peptides do not have the genetic material and activity of the influenza virus, do not cause side effects induced by the genetic material of the influenza virus, and there is no possibility that the inactivated or attenuated vaccine may be renatured in humans. 3.
  • the antibody response level is 10-20 times that of the specific neutralizing antibody induced by the corresponding subunit vaccine.
  • the amount of antibody at the position is 10-240 ⁇ g / ml serum. 4.
  • This technology will have a significant impact on world preventive medicine research, it will revolutionize the technology for preparing influenza virus vaccines, and it will bring huge economic and social benefits.
  • Example 1 Preparation of a synthetic peptide influenza vaccine based on the influenza virus membrane protein HA, including the following steps:
  • He Ij uses MBS (m-maleimidobenzoyl-N-hydroxy succinimide ester) to couple the polypeptide with the carrier protein BSA;
  • a vaccine is prepared by coupling the conjugate with an aluminum adjuvant.
  • Example 2 The preparation of a multiple-synthetic peptide influenza vaccine based on the influenza virus membrane protein HA, including the following steps:
  • An influenza virus vaccine is prepared by mixing three kinds of conjugates with aluminum adjuvant, wherein the proportion of the three kinds of conjugates can be performed according to the mutation probability of each point obtained by statistical processing in different years. Adjustment.
  • Example 3 The preparation of a multiple-synthetic peptide influenza vaccine based on the influenza virus membrane protein HA, including the following steps:
  • a peptide vaccine is prepared by mixing 5 kinds of conjugates with aluminum adjuvant, wherein the ratio of the proportions of the 3 kinds of conjugates can be adjusted according to the mutation probability of each point obtained by statistical processing in different years. .
  • Example 4 The preparation of a multi-synthetic peptide influenza vaccine based on the influenza virus M2 protein includes the following steps:
  • An influenza vaccine is prepared by mixing 4 kinds of conjugates with aluminum adjuvant, wherein the proportion of the 4 kinds of conjugates can be adjusted according to the mutation probability of each point obtained by statistical processing in different years. .
  • Example 5 Preparation of a multi-partite synthetic peptide influenza vaccine based on the influenza virus membrane protein HA and M2 protein, including the following steps
  • An influenza vaccine is prepared by mixing 5 kinds of conjugates with aluminum adjuvant, wherein the proportion of the 5 kinds of conjugates can be adjusted according to the mutation probability of each point obtained by statistical processing in different years. .
  • Example 6 The preparation of a single epitope influenza vaccine based on the influenza virus membrane protein HA includes the following steps:
  • a single epitope influenza vaccine is prepared by coupling the conjugate with an aluminum adjuvant.
  • Example 7 The preparation of a single epitope-multiple influenza vaccine based on the influenza virus membrane protein HA, including the following steps:
  • the four kinds of conjugates are mixed with aluminum adjuvant to prepare a polyvalent epitope vaccine for influenza virus.
  • the proportions of the four kinds of conjugates can be statistically processed according to different years. The point's mutation probability is adjusted.
  • Example 8 Preparation of a multi-epitope influenza virus epitope vaccine based on the influenza virus membrane proteins HA and M2 proteins, including the following steps
  • Example 9 Preparation of a multi-epitope influenza virus epitope vaccine based on influenza virus T cell epitopes, including the following steps
  • Example 10 Preparation of a single epitope-multiple influenza vaccine based on the neutralizing epitopes on influenza virus membrane proteins HA and M2 proteins and influenza virus T cell epitopes, including the following steps
  • the 8 kinds of conjugates are mixed and mixed with aluminum adjuvant to prepare an influenza vaccine, wherein the proportion of the 8 kinds of conjugates can be adjusted according to the mutation probability of each point obtained by statistical processing in different years. .
  • Example 11 The preparation of a multi-epitope-multiple influenza vaccine based on the influenza virus membrane proteins HA and M2 protein neutralizing epitopes and T cell epitopes, comprising the following steps:
  • An influenza vaccine is prepared by mixing 4 kinds of conjugates with aluminum adjuvant, wherein the proportion of the 4 kinds of conjugates can be adjusted according to the mutation probability of each point obtained by statistical processing in different years. .
  • Example 12 Preparation of an anti-mutated multi-epitope-multiple epitope influenza vaccine based on the influenza virus membrane proteins HA and M2 protein neutralizing epitopes and T cell epitopes, including the following steps
  • An influenza vaccine is prepared by mixing 4 kinds of conjugates with aluminum adjuvant.
  • the share ratios of the four types of conjugates can be adjusted according to the variation probability of each point obtained by statistical processing in different years.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Virology (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Microbiology (AREA)
  • Pulmonology (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Oncology (AREA)
  • Communicable Diseases (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

L'invention concerne un vaccin contre le virus de la grippe renfermant au moins un polypeptide épitope combiné à une protéine porteuse ou un polypeptide porteur formant des conjugués, chacun contenant un épitope de neutralisation ou un épitope de mutation se trouvant sur la protéine transmembranaire HA ou M2 du virus de la grippe ainsi que des polypeptides épitopes de lymphocytes T. Le procédé de préparation du vaccin de cette invention consiste à 1) synthétiser de manière artificielle des polypeptides contenant des épitopes de neutralisation ou des épitopes de mutation de la région antigénique se trouvant sur la protéine transmembranaire HA ou M2 du virus de la grippe et des épitopes de lymphocytes T ; 2) combiner les polypeptides à la protéine porteuse ou au polypeptide porteur afin de former des conjugués distincts ; 3) formuler un vaccin contre le virus de la grippe en combinant lesdits conjugués à l'aide d'un adjuvant acceptable. Cette invention a le mérite de lutter efficacement contre les mutations du vaccin contre le virus de la grippe, d'être rentable tout en ayant une vaste portée sociale.
PCT/CN2001/001218 2000-08-10 2001-08-03 Vaccin contre le virus de la grippe et son procede de preparation WO2002026252A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2002223398A AU2002223398A1 (en) 2000-08-10 2001-08-03 A vaccine for influenza virus and its preparation

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
CN00121290.7 2000-08-10
CNB001212931A CN1163271C (zh) 2000-08-10 2000-08-10 一种合成肽流感疫苗的制备方法
CN00121293.1 2000-08-10
CN 00121290 CN1338307A (zh) 2000-08-10 2000-08-10 一种流感病毒表位疫苗及其制备方法

Publications (1)

Publication Number Publication Date
WO2002026252A1 true WO2002026252A1 (fr) 2002-04-04

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PCT/CN2001/001218 WO2002026252A1 (fr) 2000-08-10 2001-08-03 Vaccin contre le virus de la grippe et son procede de preparation

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AU (1) AU2002223398A1 (fr)
WO (1) WO2002026252A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1603590A4 (fr) * 2003-03-07 2008-08-27 Merck & Co Inc Vaccin contre le virus de la grippe
US8444995B2 (en) 2006-02-07 2013-05-21 PepTCell, Ltd. Peptide sequences and compositions
CN110290805A (zh) * 2017-01-03 2019-09-27 埃默杰克斯疫苗控股有限公司 通用流感疫苗组合物

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993020846A1 (fr) * 1992-04-17 1993-10-28 Yeda Research And Development Co. Ltd Vaccins contre la grippe recombines
WO1996003145A1 (fr) * 1994-07-26 1996-02-08 Connaught Laboratories, Inc. Conjugues de sous-unites de virus de la grippe
US5591823A (en) * 1991-12-11 1997-01-07 American Home Products Corporation Expression of specific immunogens using viral antigens
WO2000032228A2 (fr) * 1998-11-30 2000-06-08 Yeda Research And Development Co. Ltd. Vaccin a base de peptides contre la grippe

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5591823A (en) * 1991-12-11 1997-01-07 American Home Products Corporation Expression of specific immunogens using viral antigens
WO1993020846A1 (fr) * 1992-04-17 1993-10-28 Yeda Research And Development Co. Ltd Vaccins contre la grippe recombines
WO1996003145A1 (fr) * 1994-07-26 1996-02-08 Connaught Laboratories, Inc. Conjugues de sous-unites de virus de la grippe
WO2000032228A2 (fr) * 1998-11-30 2000-06-08 Yeda Research And Development Co. Ltd. Vaccin a base de peptides contre la grippe

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1603590A4 (fr) * 2003-03-07 2008-08-27 Merck & Co Inc Vaccin contre le virus de la grippe
US8444995B2 (en) 2006-02-07 2013-05-21 PepTCell, Ltd. Peptide sequences and compositions
US8475802B2 (en) 2006-02-07 2013-07-02 Pep T cell Limited Peptide sequences and compositions
US9446116B2 (en) 2006-02-07 2016-09-20 Peptcell Limited Peptide sequences and compositions
US9889191B2 (en) 2006-02-07 2018-02-13 Peptcell Limited Peptide sequences and compositions
US10279032B2 (en) 2006-02-07 2019-05-07 Peptcell Limited Peptide sequences and compositions
US10335480B2 (en) 2006-02-07 2019-07-02 Peptcell Limited Peptide sequences and compositions
US10765734B2 (en) 2006-02-07 2020-09-08 Peptcell Limited Influenza peptides and compositions
US11439702B2 (en) 2006-02-07 2022-09-13 Peptcell Limited Influenza peptides and compositions
CN110290805A (zh) * 2017-01-03 2019-09-27 埃默杰克斯疫苗控股有限公司 通用流感疫苗组合物

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Publication number Publication date
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