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NL2034221B1 - Anti-h1n1 subtype influenza virus nanoparticles based on self-assembled ferritin, preparation method and application thereof - Google Patents

Anti-h1n1 subtype influenza virus nanoparticles based on self-assembled ferritin, preparation method and application thereof Download PDF

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NL2034221B1
NL2034221B1 NL2034221A NL2034221A NL2034221B1 NL 2034221 B1 NL2034221 B1 NL 2034221B1 NL 2034221 A NL2034221 A NL 2034221A NL 2034221 A NL2034221 A NL 2034221A NL 2034221 B1 NL2034221 B1 NL 2034221B1
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signal peptide
influenza virus
nanoparticles
seq
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Wang Mingxuan
Wan Xuehua
Wan Yan
Li Junwei
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The Second Hospital Of Nanjing Nanjing Hospital Affiliated To Nanjing Univ Of Traditional Chinese Me
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
    • C07K16/1018—Orthomyxoviridae, e.g. influenza virus
    • 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
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/42—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum viral
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/44—Antibodies bound to carriers
    • 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
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y40/00—Manufacture or treatment of nanostructures
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
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    • C07K14/315—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Streptococcus (G), e.g. Enterococci
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    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
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    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • C07K2317/00—Immunoglobulins specific features
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    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/569—Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
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    • C07K2319/735—Fusion polypeptide containing domain for protein-protein interaction containing a domain for self-assembly, e.g. a viral coat protein (includes phage display)
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Claims (8)

CONCLUSIES
1. Nanodeeltjes van anti-H1N1 subtype influenzavirus op basis van zelf-geassembleerd ferritine, waarbij — de nanodeeltjes worden gevormd door een signaalpeptide-ST-FE fusie-eiwit en een signaalpeptide-SC-HA-VHH fusie-eiwit in vitro aan elkaar te koppelen; — het signaalpeptide-ST-FE fusie-eiwit wordt verkregen door een signaalpeptide-SpyTag als weergegeven in SEQ ID NO: 3 te verbinden met een N-uiteinde van een subeenheid van een ferritine-monomeer; en — het signaalpeptide-SC-HA-VHH fusie-eiwit wordt verkregen door het N-uiteinde van een nano-antilichaam tegen hemagglutinine-eiwit van influenza type A (H1N1) subtype virus te verbinden met een signaalpeptide-SpyCatcher, zoals weergegeven in SEQ ID NO: 4.
2. De nanodeeltjes van anti-H1N1 subtype influenzavirus op basis van zelf-geassembleerd ferritine volgens conclusie 2, waarbij — de aminozuursequentie van de signaalpeptide-SpyTag en het N-uiteinde van het verbindende peptide van de subeenheid van het ferritine-monomeer is weergegeven in SEQ ID NO: 5; — de aminozuursequentie van het N-uiteinde van het nano-antilichaam tegen het hemagglutinine-eiwit van influenza type A (H1N1) subtype virus en het verbindende peptide van het signaalpeptide-SpyCatcher is weergegeven in SEQ ID NO: 5.
3. De nanodeeltjes van anti-H1N1 subtype influenzavirus op basis van zelf-geassembleerd ferritine volgens conclusie 1, waarbij — de aminozuursequentie van het signaalpeptide-ST-FE fusie-eiwit is weergegeven in SEQ ID NO: 1; en — de aminozuursequentie van het signaalpeptide-SC-HA-VHH fusie-eiwit is weergegeven in SEQ ID NO: 2.
4. De nanodeeltjes van anti-H1N1 subtype influenzavirus op basis van zelf-geassembleerd ferritine volgens conclusie 1, waarbij de nucleotidevolgorde die codeert voor het signaalpeptide-ST-FE fusie-eiwit is weergegeven in SEQ ID NO: 10.
5. De nanodeeltjes van anti-H1N1 subtype influenzavirus op basis van zelf-geassembleerd ferritine volgens conclusie 1, waarbij de nucleotidevolgorde die codeert voor het signaalpeptide-SC-HA-VHH fusie-eiwit is weergegeven in SEQ ID NO: 11.
6. Een werkwijze voor het bereiden van de nanodeeltjes van anti-H1N1 subtype influenzavirus op basis van zelf-geassembleerd ferritine volgens conclusie 3, die de volgende stappen omvat: — het construeren van genen die respectievelijk coderen voor fusie-eiwitten zoals weergegeven in SEQ ID NO: 1 en SEQ ID NO: 2, vervolgens het overbrengen van de geconstrueerde genen in vectoren om expressieplasmiden te construeren, en vervolgens het transfecteren om de expressie van fusie-eiwitten te induceren om twee soorten fusie- eiwitten te verkrijgen; en — het in vitro verbinden van de twee in stap (1) verkregen fusie-eiwitten ter verkrijging van de anti-H1N1 nanodeeltjes van het subtype influenzavirus op basis van zelf- geassembleerd ferritine.
7. De werkwijze voor het bereiden volgens conclusie 8, waarbij in stap (2) de omstandigheden van het in vitro verbinden zijn: 4°C, en 16 uur.
8. Toepassing van de nanodeeltjes van anti-H1N1 subtype influenzavirus op basis van zelf- geassembleerd ferritine volgens willekeurig welke van conclusies 1 - 5 bij de bereiding van geneesmiddelen voor het behandeling van griep van het subtype H1N1.
NL2034221A 2022-05-20 2023-02-24 Anti-h1n1 subtype influenza virus nanoparticles based on self-assembled ferritin, preparation method and application thereof NL2034221B1 (en)

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