WO2020100990A1 - 新型インフルエンザウイルス由来ヘマグルチニンタンパク質遺伝子を有するＤＩｓ株由来組換えワクシニアウイルス - Google Patents

新型インフルエンザウイルス由来ヘマグルチニンタンパク質遺伝子を有するＤＩｓ株由来組換えワクシニアウイルス Download PDF

Info

Publication number
WO2020100990A1
WO2020100990A1 PCT/JP2019/044737 JP2019044737W WO2020100990A1 WO 2020100990 A1 WO2020100990 A1 WO 2020100990A1 JP 2019044737 W JP2019044737 W JP 2019044737W WO 2020100990 A1 WO2020100990 A1 WO 2020100990A1
Authority
WO
WIPO (PCT)
Prior art keywords
virus
avian influenza
recombinant vaccinia
influenza
vaccinia virus
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/JP2019/044737
Other languages
English (en)
French (fr)
Japanese (ja)
Inventor
文彦 安井
小原　道法
賢三郎 山地
伊藤　靖
一誠 小笠原
石井　孝司
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Tokyo Metropolitan Institute of Medical Science
National Institute of Infectious Diseases
Shiga University of Medical Science NUC
Original Assignee
Tokyo Metropolitan Institute of Medical Science
National Institute of Infectious Diseases
Shiga University of Medical Science NUC
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
Application filed by Tokyo Metropolitan Institute of Medical Science, National Institute of Infectious Diseases, Shiga University of Medical Science NUC filed Critical Tokyo Metropolitan Institute of Medical Science
Priority to JP2020556172A priority Critical patent/JPWO2020100990A1/ja
Priority to EP19884182.7A priority patent/EP3888677A4/en
Priority to US17/293,378 priority patent/US20230174586A1/en
Priority to CN201980074035.0A priority patent/CN113227360A/zh
Publication of WO2020100990A1 publication Critical patent/WO2020100990A1/ja
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • 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
    • 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
    • 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
    • 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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • 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
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/525—Virus
    • A61K2039/5256—Virus expressing foreign proteins
    • 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
    • A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
    • 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
    • A61K2039/70—Multivalent vaccine
    • 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
    • C12N2710/24011—Poxviridae
    • C12N2710/24111—Orthopoxvirus, e.g. vaccinia virus, variola
    • C12N2710/24122—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
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/24011—Poxviridae
    • C12N2710/24111—Orthopoxvirus, e.g. vaccinia virus, variola
    • C12N2710/24141—Use of virus, viral particle or viral elements as a vector
    • C12N2710/24143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • 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
    • C12N2710/24011—Poxviridae
    • C12N2710/24111—Orthopoxvirus, e.g. vaccinia virus, variola
    • C12N2710/24171—Demonstrated in vivo effect
    • 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
    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
    • C12N2760/16122—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
    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
    • C12N2760/16134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • 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
    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
    • C12N2760/16141—Use of virus, viral particle or viral elements as a vector
    • C12N2760/16143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • 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
    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
    • C12N2760/16171—Demonstrated in vivo effect

Definitions

  • the present invention relates to a preventive and therapeutic agent for H7 subtype avian influenza virus infection, which is the cause of the development of novel influenza.
  • the present invention relates to a recombinant vaccinia virus DIs strain capable of expressing the hemagglutinin (HA) protein gene of H7 subtype avian influenza virus, and a preventive and therapeutic drug for novel influenza containing the recombinant vaccinia virus DIs strain. ..
  • Seasonal influenza has been circulating in winter every year, and it has been reported that 10% to 20% of the population affected.
  • the pathogen is A / H1N1, A / H3N2, or influenza B virus, but its antigenicity changes little by little every year, so it is necessary to manufacture a vaccine adapted to the epidemic prediction strain.
  • examples of new influenza include H5N1 highly pathogenic avian influenza, H7N9 avian influenza, and H7N7 highly pathogenic avian influenza.
  • an outbreak of H7N9 avian influenza virus in humans has been reported in China since 2013, and at least 1564 cases of infection and 612 deaths have been reported so far.
  • the 2016-2017 season has the largest infection scale compared to the past four seasons, and there is concern that the epidemic will spread further in the future.
  • candidate strains for vaccine production are being selected and stored in multiple countries, including the United States and the United Kingdom. A system is in place to start manufacturing.
  • the vaccine to be manufactured is a whole-particle inactivated vaccine, and therefore requires multiple vaccinations, and therefore it takes time to induce sufficient immunity.
  • the hemagglutinin (HA) protein sequence of the H7N9 avian influenza virus contains a sequence that causes activation of suppressive T cells (Treg) (Non-patent document 1), and the H7HA protein The ability to induce specific antibodies may be diminished.
  • H1N1 (2009) Considering the temporal transition of pandemic influenza virus during a global pandemic, the first wave of domestic epidemics has started in each country 2-3 months after the epidemic in the country where it occurred. Therefore, it is necessary to develop a vaccine that can respond to the H7 subtype influenza virus earlier.
  • neuraminidase which is important for the emergence of influenza virus
  • N neuraminidase
  • an intravenous NA inhibitor has already been developed.
  • Avigan a polymerase inhibitor of influenza virus, was approved for manufacturing and sales in Japan in March 2014, but will be manufactured and supplied at the request of the Minister of Health, Labor and Welfare. It is subject to conditions such as not being sold to the general public.
  • the live vaccine is one of the most effective.
  • the development of an attenuated vaccine of an emerging virus requires a very long period of time, and it is considered that this also applies to H7 subtype influenza. Therefore, the inventors of the present invention, as a result of earnest research, make full use of genetic engineering techniques to develop a recombinant vaccinia virus (RVV) that expresses the H7 subtype HA protein gene using vaccinia virus as a mother as a live influenza vaccine. I arrived.
  • RVV recombinant vaccinia virus
  • Non- Patent Document 2 a recombinant vaccinia virus (RVV) for rabies virus or for Lindapest, in field tests and the like, has been demonstrated to exhibit an excellent infection preventive effect.
  • SARS severe acute respiratory syndrome
  • Patent Document 1 a recombinant vaccinia virus having a cDNA of SARS-CoV known as a pathogen thereof (Patent Document 1), and have an excellent preventive effect and re-creation. It has been confirmed that the preparation can be administered (Non-Patent Document 3).
  • Patent Document 2 Furthermore, against H5N1HPAIV, we have also succeeded in producing a recombinant vaccinia virus having the cDNA of HA of H5N1HPAIV, and it has been confirmed that it is a formulation showing an excellent preventive effect (Patent Document 2 and non-patent document 2). Patent Document 4).
  • An embodiment of the present invention is as follows.
  • a recombinant vaccinia virus comprising an expression promoter and all or part of a cDNA encoding a hemagglutinin protein derived from an H7 subtype avian influenza virus in the genome of the vaccinia virus DIs strain.
  • H7 subtype avian influenza virus is H7N9 avian influenza virus or H7N7 highly pathogenic avian influenza virus.
  • ⁇ 3> The recombinant vaccinia virus according to ⁇ 1> or ⁇ 2>, wherein the cDNA encoding the hemagglutinin protein is the DNA of (a) to (h) below.
  • ⁇ 4> A pharmaceutical composition comprising the recombinant vaccinia virus according to any of ⁇ 1> to ⁇ 3>).
  • ⁇ 5> The pharmaceutical composition according to ⁇ 4>, which is a preventive agent for H7 subtype avian influenza.
  • ⁇ 6> The pharmaceutical composition according to ⁇ 4>, which is a therapeutic agent for H7 subtype avian influenza.
  • ⁇ 7> The pharmaceutical composition according to ⁇ 5> or ⁇ 6>, wherein the H7 subtype avian influenza is H7N9 avian influenza or H7N7 highly pathogenic avian influenza.
  • ⁇ 8> The pharmaceutical composition according to any one of ⁇ 4> to ⁇ 7>, which is for intradermal administration.
  • a preparation for intradermal administration which comprises the recombinant vaccinia virus according to any one of ⁇ 1> to ⁇ 3>.
  • the preparation for intradermal administration according to ⁇ 9> which is a preventive agent for H7 subtype avian influenza.
  • the preparation for intradermal administration according to ⁇ 9> which is a therapeutic agent for H7 subtype avian influenza.
  • ⁇ 12> The preparation for intradermal administration according to ⁇ 10> or ⁇ 11>, wherein the H7 subtype avian influenza is H7N9 avian influenza or H7N7 highly pathogenic avian influenza.
  • influenza vaccine containing the recombinant vaccinia virus according to any one of ⁇ 1> to ⁇ 3>.
  • ⁇ 14> The influenza vaccine according to ⁇ 13>, wherein the influenza is H7 subtype avian influenza.
  • ⁇ 16> The influenza vaccine according to any of ⁇ 13> to ⁇ 15>, which is for intradermal administration.
  • ⁇ 17> A method for preventing and / or treating H7 subtype avian influenza, which comprises administering an effective amount of the recombinant vaccinia virus according to any one of ⁇ 1> to ⁇ 3> to a subject.
  • ⁇ 18> The method according to ⁇ 17>, wherein the H7 subtype avian influenza is H7N9 avian influenza or H7N7 highly pathogenic avian influenza.
  • ⁇ 19> The method according to ⁇ 17> or ⁇ 18>, wherein the administration is intradermal administration.
  • ⁇ 20> The method according to any one of ⁇ 17> to ⁇ 19>, which comprises administering in combination with a vaccine against another subtype influenza virus.
  • a mixed vaccine comprising the recombinant vaccinia virus according to any one of ⁇ 1> to ⁇ 3> and one or more recombinant vaccinia viruses derived from another subtype influenza virus.
  • ⁇ 23> Use of the recombinant vaccinia virus according to any one of ⁇ 1> to ⁇ 3> in the production of influenza vaccine.
  • FIG. 1 shows the production of recombinant vaccinia virus (RVV) (specifically rDIs-H7 HA) derived from the DIs strain having the HA protein gene of H7 subtype avian influenza virus using homologous recombination technology.
  • RVV vaccinia virus
  • FIG. 2 is a photograph of a PVDF membrane showing the results of confirming the expression of the HA protein of RVV (rDIs-H7 HA) by Western blotting.
  • FIG. 3 shows a single RVV (H7 subtype avian influenza virus vaccine: rDIs-H7 HA) from a DIs strain having a HA protein gene derived from the H7N9 avian influenza virus against BALB / c mouse attack infection with the H7N9 avian influenza virus. It is a figure which shows the examination result of the immunity induction effect by infection and the enhancement effect of the infection defense ability.
  • Fig. 4 shows a method of skin scraping of RVV (H7 subtype HA gene expression vaccine: rDIs-H7 HA) derived from DIs strain having H7N9 avian influenza virus-derived HA protein gene against attack infection of H7N9 avian influenza virus to cynomolgus monkeys.
  • FIG. 6 is a diagram showing the results of examination of the immunity-inducing effect and the enhancing effect on the infection-protecting ability, and the immediate effect by the inoculation.
  • FIG. 5 is a diagram showing the results of examination of the virus elimination effect 7 days after the H7N9 avian influenza virus challenge infection in cynomolgus monkeys inoculated with the H7 subtype HA gene expression vaccine (rDIs-H7 HA).
  • FIG. 6 is a diagram showing lung pathological findings and lung weight 7 days after challenge with H7N9 avian influenza virus in cynomolgus monkeys inoculated with the H7 subtype HA gene expression vaccine (rDIs-H7 HA).
  • FIG. 5 is a diagram showing the results of examination of the virus elimination effect 7 days after the H7N9 avian influenza virus challenge infection in cynomolgus monkeys inoculated with the H7 subtype HA gene expression vaccine (rDIs-H7 HA).
  • FIG. 7 shows a single dose of RVV (H7 subtype HA gene expression vaccine: rDIs-H7 HA) derived from a DIs strain having a H7N9-derived HA protein gene against the challenge of BALB / c mice with H7N7 highly pathogenic avian influenza virus. It is a figure which shows the examination result of the infection protection effect by inoculation.
  • FIG. 8 Examination of immediate effect of DIV strain-derived RVV (H7 subtype HA gene expression vaccine: rDIs-H7 HA) against H7N9 avian influenza virus challenge infection in BALB / c mice. It is a figure which shows a result.
  • Figure 9 Intradermal of monovalent vaccine of RVV (H7 subtype HA gene expression vaccine: rDIs-H7 HA) derived from DIs strain having H7N9 avian influenza virus-derived HA protein gene against attack infection of H7N9 avian influenza virus to cynomolgus monkeys. Inoculation using the inoculation method and skin scraping method, and four types of mixed vaccines (rDIs-H1, -H3, -H5 -H7 HAs) that also include vaccines against H1, H3, and H5 subtypes in addition to influenza H7 influenza It is a figure which shows the examination result about the immunity induction effect by the intracutaneous inoculation, and the enhancing effect of the infection defense ability.
  • RVV H7 subtype HA gene expression vaccine
  • Figure 10 shows that H7 subtype HA gene expression vaccine (rDIs-H7 HA) monovalent vaccine in the cynomolgus monkey was inoculated using the intradermal or skin scraping method.
  • H1, H3, H5 Intradermal inoculation of four mixed vaccines (rDIs-H1, -H3, -H5 -H7 HAs), including vaccines against type I viruses, or lung pathological findings and viruses in lung tissue 7 days after H7N9 avian influenza virus attack infection It is a figure which shows an infectious titer.
  • the present inventors have a high safety inoculation history because it is non-proliferative in mammalian cells, and has a proven record of inoculation in humans.
  • RVV recombinant vaccinia virus
  • vaccinia virus DIs strain is a highly attenuated vaccinia virus strain isolated by subculturing vaccinia virus Dalian strain (DEI strain) in chicken egg embryos (Tagaya I et al., A new mutant of dermovaccinia). virus. Nature, 1961, vol.192, p.1187-1188).
  • This DIs strain can grow in fetal chicken fibroblasts (CEF), but it does not grow in other mammalian cells and is highly safe.
  • the vaccinia virus LC16m8 strain is a safe and effective attenuated vaccine strain that was approved by the Ministry of Health and Welfare in 1975 as a smallpox vaccine and has been inoculated to over 50,000 infants.
  • the weak toxicity of the LC16m8 strain is derived from the fact that the B5R protein gene of the LC16mO strain, which is considered to be involved in cell adsorption / infection, has a single base deletion, and therefore the full-length B5R protein is not expressed (Morikawa S ., Et al.
  • the rDIs-H7 HA inoculated individual showed a good onset protective effect against H7N9 avian influenza virus infection and H7N7 avian influenza virus infection. Furthermore, even when infected with H7N9 avian influenza virus one week or two weeks after vaccination, the symptoms could be significantly attenuated, and an immediate effect was also confirmed. From the above results, rDIs-H7HA can be a vaccine for H7 subtype influenza that can induce immunity immediately.
  • rDIs-H7 HA recombinant novel influenza vaccine
  • DIs strain As described above, the present inventor has established a recombinant novel influenza vaccine (rDIs-H7 HA) using the DIs strain as a parent.
  • RDIs-H7 HA single-vaccinated mice showed transient weight loss after challenge with A / Anhui / 1/2013 (H7N9) influenza virus, but the body weight recovered rapidly thereafter with 100% survival rate. Is shown (FIG. 3).
  • the rDIs-H7 HA twice-inoculated group showed a rapid antipyretic effect (Fig. 4) and various places (conjunctiva, nasal cavity, oral cavity) in comparison with the non-vaccinated group (DIs inoculated group).
  • RVV derived from the DIs strain expressing the influenza virus HA protein gene was lethal by single inoculation of 1x10 7 PFU of RVV as a vaccine for H7 subtype influenza virus (rDIs-H7 HA).
  • Inhibition of body weight loss was also shown against H7 influenza virus attack, with a remarkable survival-preventing effect with a survival rate of 80% when vaccinated one week ago and 100% when vaccinated two weeks ago.
  • HA hemagglutinin
  • Treg suppressive T cells
  • the HA protein gene of H7 subtype avian influenza virus for example, H7N9 avian influenza virus of SEQ ID NO: 1 has already been assigned a predetermined accession number (Accession No. EPI439507) in the influenza virus gene database (GISAID; Global Initiative on Sharing All Influenza Data). ) Is registered.
  • the gene contained in the recombinant vaccinia virus of the present invention that is, all of the cDNA encoding the HA protein of the H7N9 avian influenza virus H7 subtype avian influenza virus (including the mutant sequence) or a part thereof is a normal gene. It can be obtained by an engineering method. For example, a nucleic acid synthesis method using a DNA synthesizer generally used as a genetic engineering method can be used.
  • a primer specific to each gene is designed, and a PCR method in which the gene sequence is amplified using a PCR device, or a gene amplification method using a cloning vector Can be used.
  • the above method can be performed by referring to “Molecular Cloning, A Laboratory Manual (4th edition)” (Cold Spring Harbor Laboratory Press (2012)) and the like. Known methods can be used to purify the obtained PCR product.
  • mutant sequences in particular, mutation substitution type DNA, for example, the above-mentioned ⁇ Molecular Cloning, A Laboratory Laboratory (4th edition) ”(Cold Spring Harbor Laboratory Laboratory Press (2012)) and“ Current Protocols in Molecular Biology, John Wiley & Sons (1987-1997) "and the like, can be prepared according to the site-specific displacement induction method.
  • known methods such as the Kunkel method and the Gapped duplex method can be used to prepare using a mutation-introducing kit that utilizes the site-directed mutagenesis method.
  • a mutation-introducing kit that utilizes the site-directed mutagenesis method.
  • QuickChangeTM Site- Directed Mutagenesis Kit manufactured by Agilent Technologies
  • GeneTailorTM Site-Directed Mutagenesis System manufactured by Invitrogen
  • TaKaRa Site-Directed Mutagenesis System are preferred. Be done.
  • the H7 subtype avian influenza virus HA protein gene inserted into the above plasmid can be used as the template for the above PCR.
  • the HA protein gene of H7N9 avian influenza virus can be used as a template for the above PCR.
  • H7 subtype highly pathogenic avian influenza virus more preferably, using the cDNA of the HA protein gene of H7N9 avian influenza virus as a template, by performing PCR using each gene-specific primer, H7 subtype highly pathogenic
  • the HA protein gene region of avian influenza virus, more preferably H7N9 avian influenza virus can be prepared.
  • Each of the above mutant DNAs can be obtained by chemical synthesis, or can be subjected to colony hybridization, plaque hybridization, or Southern blot using the DNA consisting of the nucleotide sequences shown in SEQ ID NO: 1 to SEQ ID NO: 2 or a fragment thereof as a probe. It can also be obtained from a cDNA library and a genomic library by a known hybridization method such as. Examples of stringent conditions in the above hybridization include 0.1 ⁇ SSC to 10 ⁇ SSC, 0.1% to 1.0% SDS, and 20 ° C. to 80 ° C. More specifically, 37 ° C. to 56 ° C. The conditions include performing prehybridization at 30 ° C.
  • DNA that encodes HA protein (mutant DNA), having 75% or more, 80% or more, 90% or more, 95% or more, 98% or more or 99% or more homology with the nucleotide sequence shown in SEQ ID NO: 2,
  • a DNA (mutant DNA) encoding the HA protein derived from the H7N9 avian influenza virus can be used.
  • the expression promoter contained in the recombinant vaccinia virus of the present invention is not limited as long as it is inserted into the gene deficient region of the vaccinia virus DIs strain, and examples thereof include vaccinia virus promoter mH5 and the like.
  • a hybrid promoter composed of a poxvirus type A inclusion body (ATI) promoter and a multiple repeating vaccinia virus 7.5 kDa protein (p7.5) early expression promoter can be used as the expression promoter.
  • This promoter can be ligated to an appropriate plasmid, and for example, pBMSF7C and pUC / DIs are known (Arch. Virol. Vol. 138, p. 315-330, 1994; see JP-A-6-237773). ).
  • the nucleotide sequence of vaccinia virus promoter mH5 that can be used in the present invention is shown in SEQ ID NO: 3.
  • DNA having activity can also be used as an expression promoter (particularly a hybrid promoter).
  • stringent conditions are the same as above.
  • “Having promoter activity” means having transcription activity of a gene encoding a structural protein or a non-structural protein.
  • the protein expressed by the above hybrid promoter can be expressed in a large amount in a form that has undergone complete sugar modification from the early stage to the late stage of vaccinia virus infection.
  • a recombinant vaccinia virus can be prepared by introducing the plasmid vector obtained by inserting the above-mentioned expression promoter and the DNA encoding the HA protein into the host vaccinia virus.
  • the introduction of the plasmid vector into the host can adopt any known method, for example, by introducing the obtained plasmid vector into an animal cell infected with the attenuated vaccinia virus DIs strain, Recombinant vaccinia virus (rDIs-H7) that causes homologous recombination in the flanking sequences of the gene deletion region and expresses the HA protein gene of H7 subtype avian influenza virus (eg, H7N9 avian influenza virus or H7N7 highly pathogenic avian influenza virus), respectively.
  • H7 subtype avian influenza virus eg, H7N9 avian influenza virus or H7N7 highly pathogenic avian influenza virus
  • inactivated vaccine The method of manufacturing inactivated vaccine is known. As described above, embryonated chicken eggs are inoculated with the influenza virus for vaccine production and propagated, and the virus purified and concentrated from chorioallantoic fluid is partially decomposed with ether and further inactivated with formalin.
  • the recombinant vaccinia virus (rDIs-H7 HA) derived from the DIs strain that was prepared has the HA protein gene of the H7 subtype avian influenza virus (eg, H7N9 avian influenza virus) inserted into the gene deletion region of the vaccinia virus DIs strain.
  • H7 subtype avian influenza virus eg, H7N9 avian influenza virus
  • HA protein derived from influenza virus shows agglutination reaction on guinea pig erythrocytes
  • HA protein derived from vaccinia virus does not agglutinate reaction on guinea pig erythrocytes.
  • RDIV is screened by infecting animal cells with rDIs-H7HA and using the agglutination reaction of guinea pig erythrocytes to plaques formed thereby as an index.
  • the target RVV may be selected from red plaques having hemagglutination activity.
  • the virus obtained from the red plaque was subjected to PCR with the HA genome gene-specific primer of H7 subtype avian influenza virus such as H7N9 avian influenza virus using the viral genome as a template, and the HA protein gene of the H7 subtype avian influenza virus. You can check the introduction of.
  • HA protein of H7 subtype avian influenza virus such as H7N9 avian influenza virus
  • H7N9 avian influenza virus can be confirmed by Western blotting using animal cells after rDIs-H7 HA infection as a sample.
  • the antibody a rabbit antiserum prepared by immunizing a HA peptide derived from the HA protein of H7N9 avian influenza virus (SEQ ID NO: 4 etc.) can be used.
  • the thymidine kinase (TK) gene region is generally used as the gene deletion region of the vaccinia virus DIs strain, but expression of TK by inserting the target gene into the TK gene region is used. It is known that the deficiency reduces RVV proliferation. Therefore, in the present invention, the gene deletion region of the vaccinia virus DIs strain is preferable as the insertion site of the target gene.
  • the preventive and therapeutic pharmaceutical composition of novel influenza and its use includes the above recombinant vaccinia virus derived from DIs strain, and a preventive drug and treatment of novel influenza (that is, H7N9 avian influenza and H7N7 highly pathogenic avian influenza) Pharmaceuticals (prophylactic and therapeutic pharmaceutical compositions) are provided.
  • the present invention also comprises administering the recombinant vaccinia virus to a patient (subject), a method for preventing and treating the novel influenza, and the use of the recombinant vaccinia virus for preventing and treating the novel influenza, It is also possible to provide use of the recombinant vaccinia virus for producing a prophylactic and therapeutic drug for the novel influenza described above.
  • the pharmaceutical composition of the present invention can be introduced into the living body by any known method, for example, injection by intramuscular, intraperitoneal, intradermal or subcutaneous injection, or inhalation from the nasal cavity, oral cavity or lung, or oral administration.
  • a recombinant vaccinia virus contained in the pharmaceutical composition of the present invention and an existing antiviral drug eg, Tamiflu, Relenza
  • the combination mode is not particularly limited, it is also possible to simultaneously administer the recombinant vaccinia virus of the present invention and an existing antiviral drug, and after administering one, a method of administering the other after a lapse of a certain time. Can also be introduced into a living body.
  • the pharmaceutical composition of the present invention comprises known pharmaceutically acceptable carriers such as excipients, fillers, binders, lubricants, buffers, isotonic agents, chelating agents, coloring agents, preservatives, and fragrances. , A flavoring agent, a sweetening agent and the like.
  • the pharmaceutical composition of the present invention includes oral administration agents such as tablets, capsules, powders, granules, pills, solutions, syrups, parenteral administration agents such as injections, external preparations, suppositories, eye drops, etc. It can be administered orally or parenterally depending on the form. Preferably, local injection into the skin, muscle, abdominal cavity, etc. is exemplified. In a preferred embodiment of the invention said pharmaceutical composition may be for intradermal administration.
  • the pharmaceutical composition of the present invention can be provided as a preparation for intradermal administration containing the recombinant vaccinia virus of the present invention.
  • the above-mentioned preparation for intradermal administration may be a preventive agent for H7 subtype avian influenza.
  • the above-mentioned preparation for intradermal administration may be a therapeutic agent for H7 subtype avian influenza.
  • the H7 subtype avian influenza may be H7N9 avian influenza or H7N7 highly pathogenic avian influenza.
  • a method for preventing and / or treating H7 subtype avian influenza which comprises administering an effective amount of the recombinant vaccinia virus of the present invention to a subject.
  • the H7 subtype avian influenza may be H7N9 avian influenza or H7N7 highly pathogenic avian influenza.
  • the administration of the recombinant vaccinia virus of the present invention may be performed intradermally.
  • the dose is appropriately selected depending on the type of active ingredient, administration route, administration subject, age, weight, sex of patient, condition and other conditions.
  • the daily dose of virus is 1,000 to 1,000,000,000 in the case of oral administration. It is about PFU (plaque forming units), preferably about 100,000 to 100,000,000 PFU, and about 100 to 1,000,000,000 PFU for parenteral, preferably about 1,000 to 100,000,000 PFU.
  • the virus can be administered once a day or in several divided doses.
  • the recombinant vaccinia virus of the present invention is used as a preventive and therapeutic vaccine for novel influenza. Further, so far, in the development of a vaccine against the H7 subtype avian influenza virus, research focusing on antibodies against the H7N9 avian influenza virus or H7N7 highly pathogenic avian influenza virus and cytotoxic T cells (CTL) has been conducted. .. Therefore, it is preferable to measure the antibody titer or cell-mediated immune activity as a vaccine in advance. In a preferred embodiment of the present invention, the above influenza preventive and therapeutic vaccines may be for intradermal administration.
  • antibody titers against the recombinant vaccinia virus produced (rDIs-H7 HA) or parental DIs strains after inoculating mice with these virus strains, serum is collected over time, serum H7N9 avian influenza virus or H7N7 can be obtained by measuring the ELISA value for HA protein of highly pathogenic avian influenza virus.
  • the recombinant vaccinia virus produced by the present inventors can induce humoral immunity against novel influenza.
  • the recombinant vaccinia virus of the present invention can be used in the production of the influenza vaccine of the present invention.
  • vaccines comprising the recombinant vaccinia virus of the invention can be administered in combination with vaccines against other subtype influenza viruses.
  • administering the above combination is, for example, administering a recombinant vaccine containing the recombinant vaccinia virus of the present invention and another subtype influenza virus-derived recombinant vaccinia virus. You may.
  • a mixed vaccine containing the recombinant vaccinia virus of the present invention and one or more other recombinant vaccinia viruses derived from subtype influenza virus.
  • the number of other subtype influenza virus-derived recombinant vaccinia viruses that can be included in the mixed vaccine of the present invention may be one or two or more.
  • the combination vaccine of the present invention may be a dual combination vaccine.
  • the combination vaccine of the present invention may be a triple combination vaccine.
  • the combination vaccine of the present invention may be a quadruple combination vaccine.
  • other subtype influenza virus-derived recombinant vaccinia viruses may be, for example, rDIs-H1, rDIsH3, and rDIs-H5, but are not limited thereto.
  • the combination vaccine of the present invention may be a four-type combination vaccine (rDIs-H1, -H3, -H5 -H7HAs).
  • the combination vaccine of the present invention can be prepared using methods known in the art.
  • Example 1 ⁇ Method> 1) Preparation of recombinant vaccinia virus derived from influenza virus hemagglutinin protein gene-derived DIs strain Hemagglutinin protein (HA) which is an antigen of H7N9 influenza virus (A / Anhui / 1/2013) artificially synthesized downstream of the vaccinia virus promoter mH5 sequence ) Genes were ligated and inserted into a plasmid vector for homologous recombination to prepare RVV (Fig. 1). At this time, when inserting the H7N9 subtype HA sequence, SphI and AsiSI sites were used as restriction enzyme sites.
  • HA Hemagglutinin protein
  • HA protein derived from influenza virus shows agglutination reaction on guinea pig erythrocytes
  • HA protein derived from vaccinia virus does not agglutinate reaction on guinea pig erythrocytes. Therefore, RVV was screened by infecting CEF with the above recombinant virus and using the agglutination reaction of guinea pig erythrocytes to plaques formed thereby as an index. Recombinant viruses of interest were selected to form red plaques with hemagglutination activity.
  • mouse anti-H7N9-HA protein monoclonal antibody was used as the primary antibody and sheep anti-mouse IgG antibody-HRP was used as the secondary antibody for H7N9 avian influenza virus.
  • a coloring reagent manufactured by Millipore (Millipore; Immobilon western) was used.
  • H7N7 highly pathogenic avian influenza virus (A / chicken / Netherlands / 2586/2003), which is also a H7 subtype but a different HA subtype virus, was also examined.
  • BALB / c mice that were vaccinated once with the recombinant vaccinia virus (rDIs-H7 HA) by the same method were used one week or two weeks after vaccination.
  • H7N9 avian influenza virus (A / Anhui / 1/2013) was challenged with infection (50 x MLD50 / individual) and the onset protection effect was examined.
  • Recombinant vaccinia virus (rDIs-H7 HA) against H7N9 avian influenza virus infection in cynomolgus monkeys (two-pronged skin abrasion method)
  • Recombinant vaccinia virus (rDIs-H7 HA) was inoculated into both upper arms of cynomolgus monkeys with 1 ⁇ 10 7 PFU using a bifurcated needle (bifurcated needle scraping method).
  • cynomolgus monkeys that had been vaccinated once with the recombinant vaccinia virus (rDIs-H7 HA) by the same method, two weeks after vaccination, the H7N9 avian influenza virus (A / Anhui / 1/2013) attack infection (3x10 6 TCID 50 / individual) was performed and the onset protection effect was examined.
  • non-vaccination group DIs inoculation group
  • rDIs-H7 HA recombinant vaccinia virus
  • rDIs-H7 HA recombinant vaccinia virus
  • the collected lung tissue was immersed in 10% buffered formalin solution to inactivate the virus and fix the tissue, and then a paraffin block of the tissue was prepared.
  • a 4 ⁇ m thin slice was prepared, attached to a slide glass, stained with hematoxylin and eosin, and analyzed for pathological findings in the lung.
  • the vaccine of the present invention in addition to H7N9 avian influenza, H7N7 highly pathogenic avian influenza It was confirmed that it was also effective (Fig. 7). Furthermore, even in the group that was vaccinated once a week or two weeks before the H7N9 avian influenza virus challenge infection, it showed a weight loss inhibitory effect, with 80% of the vaccination 1 week before, and 2 weeks before the vaccination. A prominent onset protective effect of 100% survival rate was recognized, and it was shown that it can be used as a preventive vaccine even for rapid spread of infection (Fig. 8).
  • non-vaccination group recombinant vaccinia virus (rDIs-H7 HA) twice inoculation group, and recombinant vaccinia virus (rDIs-H7 HA) once inoculation group (rapid) using the above cynomolgus monkey .
  • the conjunctiva, nasal cavity, oral cavity, and respiratory tract were examined for virus elimination, and a biphasic pattern was observed in the non-vaccinated group (DIs-inoculated group), but the recombinant vaccinia virus (rDIs-H7 In the HA) twice inoculation group and the recombinant vaccinia virus (rDIs-H7 HA) once inoculation group (rapid), an early virus elimination effect was confirmed (Fig. 5).
  • the vaccine for H7 subinfluenza virus using the recombinant vaccinia virus (rDIs-H7 HA) of the present invention is effective against H7N9 avian influenza, and can be used as a preventive vaccine for rapid spread of infection. It was shown that.
  • the DIs strain-derived RVV expressing the influenza virus HA protein gene can be used as a vaccine for the H7 subtype influenza virus (rDIs-H7 HA) by inoculating a single dose of 1x10 7 PFU of RVV, and the lethal H7N9 influenza virus and The H7N7 influenza virus also showed a rapid weight loss-suppressing effect and 100% survival rate (two weeks before administration of the vaccine) against the challenge infection. In addition, even one week before the administration of the vaccine, a survival rate of 80% was shown. In the control group (DIs inoculation group), all individuals died. Therefore, the RVV derived from the DIs strain has a sufficient effect as a vaccine for H7 subtype influenza virus, particularly an immediate effect.
  • Example 2 Protective effect of recombinant vaccinia virus (rDIs-H7 HA) against H7N9 avian influenza virus infection in cynomolgus monkeys (examination of two-stage intradermal inoculation method) Using cynomolgus monkeys, change the method of inoculating recombinant vaccinia virus (rDIs-H7 HA) from the skin scraping method to the double-needle intradermal injection method, and protect against H7N9 avian influenza virus infection by the intradermal vaccine injection method The effect was evaluated.
  • both upper arms of cynomolgus monkeys were inoculated with a monovalent vaccine of recombinant vaccinia virus (rDIs-H7 HA) by intradermal inoculation method or skin abrasion method (1x10 7 PFU), or in addition to influenza H7 influenza H1 Intradermal inoculation (1x10 7 PFU for each) of four mixed vaccines (rDIs-H1, -H3, -H5 -H7 HAs) including vaccines against H3, H3 and H5 subtype viruses, or DIs (control) Both were carried out by single inoculation.
  • H7N9 avian influenza virus A / Anhui / 1/2013 was challenged (3x10 6 TCID 50 / individual) to examine the onset protective effect.
  • Non-vaccination group (DIs inoculation group; control group), recombinant vaccinia virus (rDIs-H7 HA) skin abrasion inoculation group, recombinant vaccinia virus (rDIs-H7 HA) intradermal inoculation group using the above cynomolgus monkeys, and The temperature of the four types of mixed vaccines (rDIs-H1, -H3, -H5 -H7HAs) was intradermally inoculated, and body temperature was measured over time immediately before the challenge.
  • virus infectivity titer in lung tissue and lung pathological analysis were performed in the vaccinated and non-vaccinated cynomolgus monkeys.
  • the collected lung tissue was immersed in a 10% buffered formalin solution to inactivate the virus and fix the tissue, and then a paraffin block of the tissue was prepared.
  • a 4 ⁇ m thin slice was prepared, attached to a slide glass, stained with hematoxylin and eosin, and analyzed for pathological findings in the lung.
  • infectious virus was detected in multiple lobes of the lung, resulting in severe pneumonia with pulmonary edema.
  • the infectious virus titer was below the detection limit and pneumonia could be significantly reduced in any of the intradermal inoculation group of the monovalent vaccine, the skin abrasion group, and the four-type mixed inoculation group (FIG. 10).
  • influenza virus HA protein gene-expressing DIs strain-derived RVV of the present invention is a safe and effective vaccine against H7 subtype influenza virus. Further, it was shown that the RVV derived from the DIs strain expressing the influenza virus HA protein gene of the present invention can be used as a preventive vaccine even for rapid spread of infection.
  • Sequence number 3 Sequence of vaccinia virus promoter mH5 aaaaattgaa aataaataca aaggttcttg agggttgtgt taaattgaaa gcgagaaata 60 atcataaata 70
  • SEQ ID NO: 4 HA full-length amino acid sequence of H7N9 avian influenza virus MNTQILVFALIAIIPTNADKICLGHHAVSNGTKVNTLTERGVEVVNATETVERTNIPRICSKGKRTVDLGQCGLLGTITGPPQCDQFLEFSADLIIERREGSDVCYPGKFVNEEALRQILRESGGIDKEAMGFTYSGIRTNGATSACRRSGSSFYAEMKWLLSNTDNAAFPQMTKSYKNTRKSPALIVWGIHHSVSTAEQTKLYGSGNKLVTVGSSNYQQSFVPSPGARPQVNGLSGRIDFHWLMLNPNDTVTFSFNGAFIAPDRASFLRGKSMGIQSGVQVDANCEGDCYHSGGTIISNLPFQNIDSRAVGKCPRYVKQRSLLLATGMKNVPEIPKGRGLFGAIAGFIENGWEGLIDGWYGFRHQNAQGEGTAADYKSTQSAIDQITGKLNRLIEKTNQQFELIDNEFNEVE

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Virology (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • General Engineering & Computer Science (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Biomedical Technology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pulmonology (AREA)
  • Plant Pathology (AREA)
  • Oncology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Communicable Diseases (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Physics & Mathematics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Immunology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
PCT/JP2019/044737 2018-11-14 2019-11-14 新型インフルエンザウイルス由来ヘマグルチニンタンパク質遺伝子を有するＤＩｓ株由来組換えワクシニアウイルス Ceased WO2020100990A1 (ja)

Priority Applications (4)

Application Number Priority Date Filing Date Title
JP2020556172A JPWO2020100990A1 (https=) 2018-11-14 2019-11-14
EP19884182.7A EP3888677A4 (en) 2018-11-14 2019-11-14 RECOMBINANT VACCINE VIRUS DERIVED FROM A DIS STRAIN HAVING A NEW HEMAGGLUTININ PROTEIN GENE DERIVED FROM THE INFLUENZA VIRUS
US17/293,378 US20230174586A1 (en) 2018-11-14 2019-11-14 Strain dis-derived recombinant vaccinia virus having novel influenza virus-derived hemagglutinin protein gene
CN201980074035.0A CN113227360A (zh) 2018-11-14 2019-11-14 具有新型流感病毒来源的血凝素蛋白基因的DIs株来源的重组牛痘病毒

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2018213990 2018-11-14
JP2018-213990 2018-11-14

Publications (1)

Publication Number Publication Date
WO2020100990A1 true WO2020100990A1 (ja) 2020-05-22

Family

ID=70731871

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2019/044737 Ceased WO2020100990A1 (ja) 2018-11-14 2019-11-14 新型インフルエンザウイルス由来ヘマグルチニンタンパク質遺伝子を有するＤＩｓ株由来組換えワクシニアウイルス

Country Status (5)

Country Link
US (1) US20230174586A1 (https=)
EP (1) EP3888677A4 (https=)
JP (1) JPWO2020100990A1 (https=)
CN (1) CN113227360A (https=)
WO (1) WO2020100990A1 (https=)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117106075A (zh) * 2023-09-04 2023-11-24 华南农业大学 一种抗h7亚型流感病毒的纳米抗体及其应用

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH06237773A (ja) 1992-12-22 1994-08-30 Tonen Corp ポックスウイルスのａ型封入体（ａｔｉ）プロモーター及び前期プロモーターを含んで成る外来遺伝子発現ベクター
JP2002017370A (ja) * 2000-07-07 2002-01-22 Japan Science & Technology Corp 遺伝子組換えワクシニアウイルスワクチン
WO2006038742A1 (ja) 2004-10-08 2006-04-13 Post Genome Institute Co., Ltd. 組み換えウイルスおよびその用途
JP2013048597A (ja) * 2011-08-31 2013-03-14 Tokyo Metropolitan Institute Of Medical Science 新型インフルエンザウイルス由来ヘマグルチニンタンパク質遺伝子を有するＤＩｓ株由来組換えワクシニアウイルス
JP2016524915A (ja) * 2013-07-15 2016-08-22 ウィスコンシン アルムニ リサーチ ファンデイション Ｍｄｃｋ、ベロ細胞又は卵内で増強された複製を有する高力価の組換えインフルエンザウイルス

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1925318A1 (en) * 2006-11-20 2008-05-28 Paul-Ehrlich-Institut Recombinant modified vaccinia virus Ankara (MVA)-based vaccine for the avian flu
CN103298936A (zh) * 2010-10-15 2013-09-11 公益财团法人东京都医学综合研究所 具有新型流感病毒来源的血凝素蛋白基因的重组痘苗病毒

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH06237773A (ja) 1992-12-22 1994-08-30 Tonen Corp ポックスウイルスのａ型封入体（ａｔｉ）プロモーター及び前期プロモーターを含んで成る外来遺伝子発現ベクター
JP2002017370A (ja) * 2000-07-07 2002-01-22 Japan Science & Technology Corp 遺伝子組換えワクシニアウイルスワクチン
WO2006038742A1 (ja) 2004-10-08 2006-04-13 Post Genome Institute Co., Ltd. 組み換えウイルスおよびその用途
JP2013048597A (ja) * 2011-08-31 2013-03-14 Tokyo Metropolitan Institute Of Medical Science 新型インフルエンザウイルス由来ヘマグルチニンタンパク質遺伝子を有するＤＩｓ株由来組換えワクシニアウイルス
JP5884100B2 (ja) 2011-08-31 2016-03-15 公益財団法人東京都医学総合研究所 新型インフルエンザウイルス由来ヘマグルチニンタンパク質遺伝子を有するＤＩｓ株由来組換えワクシニアウイルス
JP2016524915A (ja) * 2013-07-15 2016-08-22 ウィスコンシン アルムニ リサーチ ファンデイション Ｍｄｃｋ、ベロ細胞又は卵内で増強された複製を有する高力価の組換えインフルエンザウイルス

Non-Patent Citations (12)

* Cited by examiner, † Cited by third party
Title
"Molecular Cloning, A Laboratory Manual", 2012, COLD SPRING HARBOR LABORATORY PRESS
ARCH. VIROL., vol. 138, 1994, pages 1987 - 1997
KITABATAKE M. ET AL.: "SARS-CoV spike protein-expressing recombinant vaccinia virus efficiently induces neutralizing antibodies in rabbits pre-immunized with vaccinia virus", VACCINE, vol. 25, 2007, pages 630 - 637, XP005798920, DOI: 10.1016/j.vaccine.2006.08.039
LIU R ET AL.: "H7N9 T-cell epitopes that mimic human sequences are less immunogenic and may induce Treg-mediated tolerance", HUM VACCIN IMMUNOTHER, vol. 11, no. 9, 2015, pages 2241 - 52
MESEDA, CA ET AL.: "Immunogenicity and Protection Against Influenza H7N3 in Mice by Modified Vaccinia Virus Ankara Vectors Expressing Influenza Virus Hemagglutinin or Neuraminidase", SCIENTIFIC REPORTS, vol. 8, no. 5364, 29 March 2018 (2018-03-29), pages 5364, XP055707233 *
MORIKAWA S. ET AL.: "An attenuated LC16m8 smallpox vaccine: analysis of full-genome sequence and induction of immune protection", J. VIROL., vol. 79, no. 18, 2005, pages 11873 - 1189
See also references of EP3888677A4
TAGAYA I ET AL.: "A new mutant of dermovaccinia virus", NATURE, vol. 192, 1961, pages 1187 - 1188
TSUKIYAMA K. ET AL.: "Development of heat-stable recombinant rinderpest vaccine", ARCH. VIROL., vol. 107, 1989, pages 225 - 235, XP000351661, DOI: 10.1007/BF01317919
YAMAJI, KENZABURO ET AL.: "2-3-11 Examination of onset defense effect of recombinant vaccinia vaccine expressing H7N9 avian influenza HA protein in cynomolgus monkeys", PROGRAMS AND ABSTRACTS OF 22ND ACADEMIC CONFERENCE OF THE JAPANESE SOCIETY FOR VACCINOLOGY; DECEMBER 8-9, 2018, vol. 22, 20 November 2018 (2018-11-20), JP, pages 118, XP009528371 *
YASUI F. ET AL.: "Sensitization with vaccinia virus encoding H5N1 hemagglutinin restore immune potential against H5N1 influenza virus", SCI. REP., vol. 6, 2016, pages 37915, XP055802741, DOI: 10.1038/srep37915
YASUI, FUMIHIKO ET AL: "1-2-09 Analysis of the onset defense mechanism by the recombinant H5- subtype HA influenza vaccine which is based on attenuated vaccinia virus DIs strain", PROGRAMS AND ABSTRACTS OF 21ST ACADEMIC CONFERENCE OF THE JAPANESE SOCIETY FOR VACCINOLOGY; DECEMBER 2-3, 2017, vol. 21, 2017, JP, pages 102, XP009528372 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117106075A (zh) * 2023-09-04 2023-11-24 华南农业大学 一种抗h7亚型流感病毒的纳米抗体及其应用

Also Published As

Publication number Publication date
CN113227360A (zh) 2021-08-06
JPWO2020100990A1 (https=) 2020-05-22
US20230174586A1 (en) 2023-06-08
EP3888677A1 (en) 2021-10-06
EP3888677A4 (en) 2022-08-17

Similar Documents

Publication Publication Date Title
KR100571479B1 (ko) Hiv에 대한 폴리엔브백신으로서 재조합 백시니아벡터의 혼합물
KR20090117697A (ko) 조류 독감용 재조합 변형된 백시니아 바이러스 안카라 (ｍｖａ)-기반 백신
WO2012053646A1 (ja) ワクシニアウイルスベクターおよびセンダイウイルスベクターからなるプライム／ブーストワクチン用ウイルスベクター
JP2013523096A (ja) インフルエンザマトリックス２タンパク質の外部ドメイン、発現システムおよびそれらの使用
EP4205762A1 (en) Improved dna vaccine for sars-cov-2
JP2024537250A (ja) インフルエンザワクチン
JP2023522154A (ja) インフルエンザワクチン
KR20230022206A (ko) 코로나바이러스 질환에 대응하는 재조합적 변형된 우두증 바이러스 앙카라 (mva) 백신
US20180161421A1 (en) Recombinant vaccinia virus having hemagglutinin protein genes derived from novel influenza viruses
JP5884100B2 (ja) 新型インフルエンザウイルス由来ヘマグルチニンタンパク質遺伝子を有するＤＩｓ株由来組換えワクシニアウイルス
WO2020100990A1 (ja) 新型インフルエンザウイルス由来ヘマグルチニンタンパク質遺伝子を有するＤＩｓ株由来組換えワクシニアウイルス
KR20250071176A (ko) 사스-코로나바이러스-2 범용 항원 폴리펩타이드, 폴리뉴클레오타이드 및 이의 용도
CN103865923B (zh) 流感病毒为载体的HAdV嵌合疫苗的制备及其应用
WO2006002594A1 (en) A recombinant canine adenovirus type-2 and the preparation method and usage thereof
KR102959050B1 (ko) 변형된 사스-코로나바이러스-2 ORF3a 폴리펩타이드 및 이의 용도
WO2022025298A1 (ja) 組換えワクシニアウイルス
KR20260056047A (ko) 변형된 사스-코로나바이러스-2 ORF3a 폴리펩타이드 및 이의 용도
JP2024503482A (ja) 複製可能アデノウイルス４型ｓａｒｓ－ｃｏｖ－２ワクチンおよびそれらの使用
HK40090677A (zh) 流感疫苗
EP4282429A1 (en) Multivalent covid-19 vaccines based on adenoviral vectors
JPWO2019069561A1 (ja) インフルエンザに対する医薬
ÇÖVEN et al. Development of an Inactivated Trivalent Influenza Vaccine with Saponin Adjuvant Isolated from Cephalaria Species: Evaluation of Immunization and Protection in Animal Models

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19884182

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2020556172

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2019884182

Country of ref document: EP

Effective date: 20210614