WO2022197209A1 - Induction d'une immunité contre le sras-cov-2 chez les enfants - Google Patents

Induction d'une immunité contre le sras-cov-2 chez les enfants Download PDF

Info

Publication number
WO2022197209A1
WO2022197209A1 PCT/RU2022/000047 RU2022000047W WO2022197209A1 WO 2022197209 A1 WO2022197209 A1 WO 2022197209A1 RU 2022000047 W RU2022000047 W RU 2022000047W WO 2022197209 A1 WO2022197209 A1 WO 2022197209A1
Authority
WO
WIPO (PCT)
Prior art keywords
seq
agent
cov
genome
utilization
Prior art date
Application number
PCT/RU2022/000047
Other languages
English (en)
Inventor
Olga Vadimovna ZUBKOVA
Tatiana Andreevna OZHAROVSKAIA
Inna Vadimovna DOLZHIKOVA
Olga Popova
Dmitrii Viktorovich SHCHEBLIAKOV
Daria Mikhailovna GROUSOVA
Alina Shahmirovna DZHARULLAEVA
Amir Ildarovich TUKHVATULIN
Natalia Mikhailovna TUKHVATULINA
Dmitrii Nikolaevich SHCHERBININ
Ilias Bulatovich ESMAGAMBETOV
Elizaveta Alexandrovna TOKARSKAYA
Andrei Gennadevich BOTIKOV
Alina Sergeevna EROKHOVA
Fatima Magomedovna Izhaeva
Natalia Anatolievna NIKITENKO
Nadezhda Leonidovna LUBENETS
Aleksandr Sergeevich SEMIKHIN
Boris Savelievich NARODITSKY
Denis Yuryevich LOGUNOV
Aleksandr Leonidovich GINTSBURG
Sergey Vladimirovich Borisevich
Vladimir Aleksandrovich Chernetsov
Evgenii Vladimirovich Kriukov
Vladimir Fedorovich Babira
Dmitrii Anatolievich KUTAEV
Svetlana Yakovlevna Loginova
Original Assignee
Federal State Budgetary Institution "National Research Centre For Epidemiology And Microbiology Named After The Honorary Academician N. F. Gamaleya" Of The Ministry Of Health Of The Russian Federation
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 Federal State Budgetary Institution "National Research Centre For Epidemiology And Microbiology Named After The Honorary Academician N. F. Gamaleya" Of The Ministry Of Health Of The Russian Federation filed Critical Federal State Budgetary Institution "National Research Centre For Epidemiology And Microbiology Named After The Honorary Academician N. F. Gamaleya" Of The Ministry Of Health Of The Russian Federation
Priority to CN202280000642.4A priority Critical patent/CN116528895A/zh
Priority to EP22713843.5A priority patent/EP4436599A1/fr
Priority to KR1020227010857A priority patent/KR20240135414A/ko
Priority to CA3156264A priority patent/CA3156264A1/fr
Priority to JP2022520203A priority patent/JP2023512381A/ja
Priority to MX2022004059A priority patent/MX2022004059A/es
Priority to ZA2022/03566A priority patent/ZA202203566B/en
Priority to IL291816A priority patent/IL291816B2/en
Publication of WO2022197209A1 publication Critical patent/WO2022197209A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • 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
    • A61K39/215—Coronaviridae, e.g. avian infectious bronchitis virus
    • 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
    • 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
    • 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
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • 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
    • A61K2039/572—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 cytotoxic response
    • 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
    • A61K2039/575—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 humoral response
    • 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
    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10341—Use of virus, viral particle or viral elements as a vector
    • C12N2710/10343—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
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/20011—Coronaviridae
    • C12N2770/20022—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
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/20011—Coronaviridae
    • C12N2770/20034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • the invention relates to biotechnology, immunology and virology.
  • the claimed agent can be used for prevention of diseases caused by severe acute respiratory syndrome virus SARS- CoV-2.
  • SARS-CoV-2 spread in Hubei province, People's Republic of China.
  • the epidemic disease caused by SARS-CoV-2 is called coronavirus disease- 19, abbreviated as COVID-19.
  • the given disease can proceed both in an asymptomatic and mild form and in a severe form that can be accompanied with sepsis and multiple organ system failure.
  • the World Health Organization declared the SARS-CoV-2-related outbreak to be a public health emergency of international concern and in March it described the spread of the disease as a pandemic.
  • July 28, 2021 over 195 million cases of illness were confirmed and 4 million people died.
  • the pharmaceutical company Pfizer in cooperation with the biotechnology company BioNTech, developed the BNT162b2 vaccine (tozinameran).
  • the given vaccine represents lipid nanoparticles with encapsulated modified mRNA encoding the SARS-CoV-2 S-protein mutant form. At the moment it is allowed to use this vaccine for adults and children aged 12 years and older (F.P. Polacketal. Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine. N Engl J Med 2020; 383: 2603-2615; www.cdc.gov/coronavirus/2019- ncov/vaccines/recommendations/adolescents.html).
  • the pharmaceutical company Modema in cooperation with the National Institute of Health (USA), developed the mRNA-1273 vaccine.
  • the active component of this vaccine is mRNA encoding SARS-CoV-2 S protein, which is surrounded with a lipid coating.
  • mRNA encoding SARS-CoV-2 S protein which is surrounded with a lipid coating.
  • SerBASE a clinical study is under way that is called KidCOVE, in which children aged 6 months - 12 years are immunized (L. A. Jackson etal. An mRNA Vaccine against SARS-CoV-2 — Preliminary Report. N Engl J Med 2020; 383:1920-1931;https://penntoday.upenn.edu/news/covid-vaccine- kids; https://clinicaltrials.gov/ct2/show/NCT04796896).
  • the University of Oxford in cooperation with the pharmaceutical company AstraZeneca, developed the ChAdOxl nCoV-19 vector vaccine (AZD1222).
  • the active component of this vaccine is chimpanzee adenovirus ChAdOxl, including the codon-optimized encoding sequence of full-length S protein of SARS-CoV-2 vims (GenBank MN908947) with the tissue plasminogen activator leader sequence.
  • the vaccination protocol includes double immunization with an interval of 28 days (M. Voysey et al. Safety and efficacy of the ChAdOxl nCoV-19 vaccine (AZD1222) against SARS-CoV-2: an interim analysis of four randomised controlled trials in Brazil, South Africa, and the UK. TheLancet. Vol. 397, Issue 10269, P99-111, 2021).
  • CanSino Company developed a vector vaccine against COVID-19, based on recombinant human adenovirus serotype 5 (Ad5) expressing SARS-CoV-2 full-length S -glycoprotein. At present, the vaccine is intended for urgent use in adults aged 18 years or older.
  • GenBankYP_009724390 (Feng-Cai Zhu et al. Immunogenicity and safety of a recombinant adenovirus type-5-vectored COVID-19 vaccine in healthy adults aged 18 years or older: a randomised, double-blind, placebo-controlled, phase 2 trial. The Lancet. Vol. 369, Issue 10249, P479-488, 2020).
  • Ad26COV2.S Ad26COVSl
  • the active component of this vaccine is recombinant adenovirus vector serotype 26 with deletion of El and E3 region, comprising the SARS-CoV-2 S-protein gene, with furin cleavage site mutation and with two proline-stabilizing mutations.
  • the vaccine may be utilized in urgent conditions for adults aged 18 years or older. Currently there are ongoing studies of vaccine utilization in teenagers and children from birth. (J. Sadoff et al.
  • Protective immunity against SARS-CoV-2 coronavirus activates several chains of the immune system. It seems that an effective vaccine against COVID-19 shall induce both humoral and cell immune response. Besides, an important element of protective immunity shall be activation of mucosal immunity (for instance, the one implemented via expression of IgA antibodies) in the nasopharynx, which is the virus penetration gate.
  • the technical task of the claimed group of inventions is creation of agents for effective induction of immune response (including mucosal immune response) against the SARS-CoV-2 virus in children aged 1 month and older.
  • the technical result consists in creation of a safe and effective agent enabling development of reactions of humoral and cell immune response against the SARS-CoV-2 virus in children aged 1 month and older.
  • a child is bom with an immature congenital and adaptive immune system that develops and acquires memory with the child's growth. From human birth till puberty the immune system undergoes several stages in its development.
  • Neonatal T-cells greatly differ from adult cells, which is a consequence of antenatal life, when the impact of nonshared antigens is significantly limited by maternal alloantigens. Therefore, the very early adaptive T-cell immunity is characterized with tolerogenic reactivity, decreased alloantigen recognition and weak responses to nonshared antigens. Newborns' B-cells greatly differ from an adult's B-cells, too.
  • Neonatal B-cells are known to have decreased TACI, BCMA and BAFF-R expression as well as decreased IgG and IgA production in response to CD40L and IL-10 (Kaur K, Chowdhury S, Greenspan NS, Schreiber JR. Decreased expression of tumor necrosis factor family receptors involved in humoral immune responses in preterm neonates. Blood. 2007 Oct 15;110(8):2948-54. doi: 10.1182/blood-2007-01-069245. Epub 2007 Jul 18. PMID: 17634409). Together, these peculiarities contribute to depression of humoral immune responses with incomplete switching of the class of immunoglobulins.
  • IgG class antibodies With the child grows, his or her contacts with the outer world increase. Gradually there is switching of immune reactions to formation of IgG class antibodies. However, the primary immune response to many antigens remains (IgM synthesis). The local immunity system still remains immature. Gradually the average blood concentration of IgG and IgM increases and reaches the level corresponding to that of adults, however the blood level of IgA still does not reach end values.
  • the virus When a child faces SARS-CoV-2, the virus, first of all, affects the mucous membranes of the respiratory tract. It means that interactions between the virus and the immune system first take place primarily on the mucous membranes of the respiratory tract and the oral cavity. Therefore, induction of mucosal immunity is an important factor influencing protective properties of a pharmaceutical agent.
  • an agent comprising a component in the form of an expression vector based on the genome of recombinant human adenovirus serotype 26 in which El and E3 site are deleted, while ORF6-Ad26 site is replaced with ORF6-Ad5 with an integrated expression cassette selected from SEQ ID NO:l, SEQ ID NO:2, SEQ ID NOG for induction of specific immunity against the severe acute respiratory syndrome virus SARS-CoV-2 in children aged 1 month and older.
  • an agent comprising a component in the form of an expression vector based on the genome of recombinant human adenovirus serotype 5 with deletion of El and E3 sites with an integrated expression cassette selected from SEQ ID NO:l, SEQ ID NOG, SEQ ID NOG for induction of specific immunity against the severe acute respiratory syndrome virus SARS-CoV-2 in children aged 1 month and older.
  • an agent comprising a combination representing component 1 in the form of an expression vector based on the genome of recombinant human adenovirus serotype 26 in which El and E3 sites are deleted, while ORF6-Ad26 site is replaced with ORF6-Ad5 with an integrated expression cassette selected from SEQ ID NO:l, SEQ ID NOG, SEQ ID NOG, and component 2 in the form of an expression vector based on the genome of recombinant human adenovirus serotype 5 with deletion of El and E3 sites with an integrated expression cassette selected from SEQ ID NO:l, SEQ ID NOG, SEQ ID NOG for induction of specific immunity against the severe acute respiratory syndrome virus SARS-CoV-2 in children aged 1 month and older.
  • an agent comprising a component in the form of an expression vector based on the genome of recombinant simian adenovirus serotype 25 with deletion of El and E3 sites with an integrated expression cassette selected from SEQ ID NO:4, SEQ ID NO:2, SEQ ID NO: 3 for induction of specific immunity against the severe acute respiratory syndrome virus SARS-CoV-2 in children aged 1 month and older.
  • an agent comprising a combination representing component 1 in the form of an expression vector based on the genome of recombinant human adenovirus serotype 26 in which El and E3 sites are deleted, while ORF6-Ad26 site is replaced with ORF6-Ad5 with an integrated expression cassette selected from SEQ ID NO:l, SEQ ID NO:2, SEQ ID NO:3, and component 2 in the form of an expression vector based on the genome of recombinant simian adenovirus serotype 25 with deletion of El and E3 sites with an integrated expression cassette selected from SEQ ID NO:4, SEQ ID NO:2, SEQ ID NO:3 for induction of specific immunity against the severe acute respiratory syndrome virus SARS-CoV-2 in children aged 1 month and older.
  • an agent comprising a combination representing component 1 in the form of an expression vector based on the genome of recombinant simian adenovirus serotype 25 with deletion of El and E3 sites with an integrated expression cassette selected from SEQ ID NO:4, SEQ ID NO:2, SEQ ID NO:3, and component 2 in the form of an expression vector based on the genome of recombinant human adenovirus serotype 5 with deletion of El and E3 sites with an integrated expression cassette selected from SEQ ID NO:l, SEQ ID NO:2, SEQ ID NO: 3 for induction of specific immunity against the severe acute respiratory syndrome virus SARS-CoV-2 in children aged 1 month and older.
  • the agent induces mucosal immune response on mucous membranes of the respiratory tract
  • the agent is prepared in a liquid or lyophilized form.
  • liquid form of the agent comprises a buffer, wt. %:
  • the reduced lyophilized form of the agent comprises a buffer, wt. %:
  • a component and/or components of the agent are intended for intranasal and/or intramuscular administration.
  • the agent is intended for administration with the dose of 5*10 9 - 5*10 10 virus particles.
  • the agent components are intended for sequential administration with an interval of over 1 week or are intended for simultaneous administration.
  • agent components can be in individual packages.
  • FIG. 1 illustrates the percentage of proliferating CD4+ (A) and CD8+ (E) T lymphocytes before immunization (Day 1) and on Day 14 of the study, after immunization of mice with the developed pharmaceutical agent Ad26-CMV-S-CoV2. Dots denote the values for each animal participating in the study. The median value is shown with a black line for each group of data. Deviations denote 95% confidence interval. The symbol ** denotes a statistically significant difference between values of Days 1 and 14 (p ⁇ 0.01, Mann- Whitney test).
  • FIG. 2 illustrates the percentage of proliferating CD4+ (A) and CD8+ (B) T lymphocytes before immunization (Day 1) and on Day 14 of the study, after immunization of mice with the developed pharmaceutical agent Ad5-CMV-S-CoV2.
  • Dots denote the values for each animal participating in the study. The median value is shown with a black line for each group of data. Deviations denote 95% confidence interval.
  • the symbols * (p ⁇ 0.05) and ** (p ⁇ 0.01) denote a statistically significant difference between values of Days 1 and 14, Mann- Whitney test.
  • FIG. 3 illustrates titers of IgG antibodies specific to RBD-domain of S ARS-Cov2 virus S protein, before vaccination (Day 1) and on Days 21, 28 and 42 of the study, after immunization of volunteers with the developed pharmaceutical agent with the dose of lxlO 10 virus particles.
  • Dots denote the values for each volunteer participating in the study.
  • the mean geometric value of the antibody titer is represented with a black line for each group of data. Deviations denote 95% confidence interval.
  • the statistically significant difference between values on Days 21, 28 and 42 is shown with a bracket above which the value p, Wilcoxon rank sum test, is shown (#### - pO.OOOl).
  • UC means uncertain differences between the given data samples.
  • the statistically significant difference between values of Days 21, 28 and 42 comparing to the values before vaccination (Day 1) was determined by Wilcoxon rank sum test (**** - pO.OOOl).
  • FIG. 4 illustrates titers of IgG antibodies specific to RBD-domain of S ARS-Cov2 virus S protein, before vaccination (Day 1) and on Days 21, 28 and 42 of the study, after immunization of volunteers with the developed pharmaceutical agent with the dose of 2xl0 10 virus particles.
  • Dots denote the values for each volunteer participating in the study.
  • the mean geometric value of the antibody titer is represented with a black line for each group of data. Deviations denote 95% confidence interval.
  • the statistically significant difference between values on Days 21, 28 and 42 is shown with a bracket above which the value p, Wilcoxon rank sum test, is shown (#### - pO.OOOl).
  • UC means uncertain differences between the given data samples.
  • the statistically significant difference between values of Days 21, 28 and 42 comparing to the values before vaccination (Day 1) was determined by Wilcoxon rank sum test (**** - p O.OOOl).
  • FIG. 5 illustrates the percentage of proliferating CD4+ (A) and CD8+ (B) T lymphocytes before immunization (Day 1) and on Day 28 of the study, after immunization of volunteers with the developed pharmaceutical agent with the dose of lxlO 10 virus particles.
  • Dots denote the values for each volunteer participating in the study. The median value is shown with a black line for each group of data. Deviations denote 95% confidence interval.
  • the symbol **** denotes the statistically significant difference between values of Days 1 and 28 (p ⁇ 0.0001, according to Wilcoxon rank sum test).
  • FIG. 6 illustrates the percentage of proliferating CD4+ (A) and CD8+ (B) T lymphocytes before immunization (Day 1) and on Day 28 of the study, after immunization of volunteers with the developed pharmaceutical agent with the dose of 2x10 10 virus particles.
  • Dots denote the values for each volunteer participating in the study. The median value is shown with a black line for each group of data. Deviations denote 95% confidence interval. The symbol **** denotes the statistically significant difference between values of Days 1 and 28 (pO.0001, according to Wilcoxon rank sum test).
  • the first stage in the development of an immunobiological agent against the severe acute respiratory syndrome virus SARS-CoV-2 was the selection of a vaccine antigen.
  • the literature search was performed which demonstrated that the coronavirus S protein was the most promising antigen for creating a candidate vaccine.
  • This is Type 1 transmembrane glycoprotein responsible for virus particles binding, fusion and entry into the cells.
  • it is an inducer of neutralizing antibodies (Liang M et al, SARS patients-derived human recombinant antibodies to S and M proteins efficiently neutralize SARS-coronavirus infectivity. BiomedEnvironSci. 2005 Dec;18(6):363-74).
  • the expression cassette SEQ ID NO:l consists of the CMV promoter, gene of SARS- CoV-2 virus S protein and polyadenylation signal.
  • the CMV promoter is the promoter of early cytomegalovirus genes that enables constitutive expression in a multitude of cell types.
  • the power of target gene expression managed by the CMV promoter varies depending on the cell type.
  • the transgene expression level under control of the CMV promoter was shown to decrease with increase of cell cultivation time due to gene expression suppression, which is related to DNA methylation [Wang W., Jia YL., Li YC., Jing CQ., Guo X., Shang XF., Zhao CP., Wang TY. Impact of different promoters, promoter mutation, and an enhancer on recombinant protein expression in CHO cells. // Scientific Reports - 2017. - Vol. 8. - P. 10416].
  • the expression cassette SEQ ID NO:2 consists of the CAG promoter, gene of SARS- CoV-2 virus S protein and polyadenylation signal.
  • the CAG-promoter is the synthetic promoter that includes the early enhancer of the CMV promoter, chicken b-actin promoter and chimeric intron (chicken b-actin and rabbit b-globin). It was experimentally shown that the transcription activity of the CAG promoter is higher than that of the CMV promoter. [Yang C.Q., Li X.Y., Li Q., Fu S.L., Li H., Guo Z.K., Lin J.T., Zhao S.T. Evaluation of three different promoters driving gene expression in developing chicken embryo by using in vivo electroporation. // Genet. Mol. Res. - 2014. - Vol. 13. - P. 1270- 1277]
  • the expression cassette SEQ ID NO:3 consists of the EF1 promoter, gene of SARS- CoV-2 virus S protein and polyadenylation signal.
  • the EF1 -promoter is the promoter of human eukaryotic translation elongation factor 1b (EF-la).
  • the promoter is constitutively active within a wide range of cell types [PMID: 28557288.
  • the EF-la promoter maintains high-level transgene expression from episomal vectors in transfected CHO-K1 cells].
  • the EF-la gene encodes the elongation factor- la, which is one of the most widely spread proteins in eukaryotic cells and is expressed in all types of mammalian cells. This EF-la promoter is often active in the cells in which virus promoters are incapable of expressing controlled genes and in cells in which virus promoters are gradually suppressed.
  • the expression cassette SEQ ID NO:4 consists of the CMV promoter, gene of SARS-CoV-2 virus S protein and polyadenylation signal.
  • Adenovirus vectors have a whole range of advantages: they are incapable of proliferating in human cells, they penetrate both in proliferating and non-proliferating cells, they are capable of inducing cell and humoral immune response, they ensure a high level of target antigen expression.
  • the authors developed variants of the agent comprising two components as well as variants comprising one component based on adenoviridae of different serotypes.
  • the immune response to the adenovirus vector part which can arise after administration of the first component of the agent or single-component agent is not boostered further on and does not influence generation of antigen-specific immune responses against the vaccine antigen in case of utilization of the two-component agent or in case of the need in repeated administration of the single-component agent as in the latter case an agent based on another adenovirus can be administered.
  • the developed agents widen the range of agents for induction of immune response against the SARS-CoV-2 coronavirus, which will enable overcoming of difficulties related to the problem of presence of pre-immunity to some serotypes of adenoviridae in a part of population.
  • an agent comprising a component in the form of an expression vector based on the genome of recombinant human adenovirus serotype 26 in which El and E3 site are deleted, while ORF6-Ad26 site is replaced with ORF6-Ad5 with an integrated expression cassette selected from SEQ ID NO:l, SEQ ID NO:2, SEQ ID NO:3 for induction of specific immunity against the severe acute respiratory syndrome virus SARS-CoV-2 in children aged 1 month and older.
  • an agent comprising a component in the form of an expression vector based on the genome of recombinant human adenovirus serotype 5 with deletion of El and E3 sites with an integrated expression cassette selected from SEQ ID NO:l, SEQ ID NO:2, SEQ ID NO: 3 for induction of specific immunity against the severe acute respiratory syndrome virus SARS-CoV-2 in children aged 1 month and older.
  • an agent comprising a combination representing component 1 in the form of an expression vector based on the genome of recombinant human adenovirus serotype 26 in which El and E3 sites are deleted, while ORF6-Ad26 site is replaced with ORF6-Ad5 with an integrated expression cassette selected from SEQ ID NO:l, SEQ ID NO:2, SEQ ID NO:3, and component 2 in the form of an expression vector based on the genome of recombinant human adenovirus serotype 5 with deletion of El and E3 sites with an integrated expression cassette selected from SEQ ID NO:l, SEQ ID NO:2, SEQ ID NO:3 for induction of specific immunity against the severe acute respiratory syndrome virus SARS-CoV-2 in children aged 1 month and older.
  • an agent comprising a component in the form of an expression vector based on the genome of recombinant simian adenovirus serotype 25 with deletion of El and E3 sites with an integrated expression cassette selected from SEQ ID NO:4, SEQ ID NO:2, SEQ ID NO: 3 for induction of specific immunity against the severe acute respiratory syndrome virus SARS-CoV-2 in children aged 1 month and older.
  • an agent comprising a combination representing component 1 in the form of an expression vector based on the genome of recombinant human adenovirus serotype 26 in which El and E3 sites are deleted, while ORF6-Ad26 site is replaced with ORF6-Ad5 with an integrated expression cassette selected from SEQ ID NO:l, SEQ ID NO:2, SEQ ID NO:3, and component 2 in the form of an expression vector based on the genome of recombinant simian adenovirus serotype 25 with deletion of El and E3 sites with an integrated expression cassette selected from SEQ ID NO:4, SEQ ID NO:2, SEQ ID NO:3 for induction of specific immunity against the severe acute respiratory syndrome virus SARS-CoV-2 in children aged 1 month and older.
  • an agent comprising a combination representing component 1 in the form of an expression vector based on the genome of recombinant simian adenovirus serotype 25 with deletion of El and E3 sites with an integrated expression cassette selected from SEQ ID NO:4, SEQ ID NO:2, SEQ ID NO:3, and component 2 in the form of an expression vector based on the genome of recombinant human adenovirus serotype 5 with deletion of El and E3 sites with an integrated expression cassette selected from SEQ ID NO:l, SEQ ID NO:2, SEQ ID NO: 3 for induction of specific immunity against the severe acute respiratory syndrome virus SARS-CoV-2 in children aged 1 month and older.
  • the agent induces mucosal immune response on mucous membranes of the respiratory tract
  • the agent is prepared in a liquid or lyophilized form.
  • liquid form of the agent comprises a buffer, wt. %:
  • the reduced lyophilized form of the agent comprises a buffer, wt. %:
  • a component and/or components of the agent are intended for intranasal and/or intramuscular administration.
  • the agent is intended for administration with the dose of 5*10 9 - 5*10 10 virus particles.
  • the agent components are intended for sequential administration with an interval of over 1 week or are intended for simultaneous administration.
  • agent components can be in individual packages.
  • the authors developed buffer variants that enable storing both in a frozen form at temperature below -18°C and in the form of a lyophilisate at temperature from +2°C to +8°C.
  • the agent also consisting of one component, can be used once.
  • Revaccination can be performed with any of the claimed agents irrespective of the agent used for vaccination.
  • the design of the plasmid construct pAd26-Ends comprising two sites homologous to the genome of human adenovirus serotype 26 (two homology arms) and the gene of resistance to ampicillin.
  • One homology arm is the beginning of the genome of human adenovirus serotype 26 (from the left inverted terminal repeat to El site) and the sequence of the virus genome comprising pIX protein.
  • the second homology arm comprises the nucleotide sequence after ORF3 E4 site to the genome end.
  • the pAd26-Ends construct was synthesized by CJSC "Evrogen" (Moscow).
  • Human adenovirus serotype 26 DNA isolated from virions was mixed with pAd26- Ends.
  • the pAd26-dlEl plasmid was obtained that comprised the genome of human adenovirus serotype 26 with deleted El site.
  • the expression cassette SEQ ID NO:l consists of the CMV promoter, gene of SARS-CoV-2 virus S protein and polyadenylation signal;
  • the expression cassette SEQ ID NO:2 consists of the CAG promoter, gene of SARS-CoV-2 virus S protein and polyadenylation signal;
  • the expression cassette SEQ ID NOG consists of the EF1 promoter, gene of SARS- CoV-2 virus S protein and polyadenylation signal.
  • constructs pArms-26-CMV-S-CoV2, pArms-26-CAG-S-CoV2, pArms-26-EFl-S- CoV2 were obtained that comprised expression cassettes SEQ ID NO:l, SEQ ID NOG or SEQ ID NOG, respectively, as well as homology arms of adenovirus serotype 26 genome.
  • each plasmid was mixed with the recombinant vector pAd26-only-null.
  • the pAd26-only-CMV-S-CoV2, pAd26-only-CAG-S-CoV2, pAd26-only-EFl-S-CoV2 plasmids were hydrolyzed with specific restriction endonucleases to delete the vector part.
  • the obtained DNA preparations were used to transfect HEK293 culture cells.
  • the expression vector was obtained that comprised the genome of recombinant human adenovirus serotype 26 in which El and E3 sites were deleted, while ORF6-Ad26 region was replaced with ORF6-Ad5 with the integrated expression cassette selected from SEQ ID NO:l, SEQ ID NOG, SEQ ID NOG.
  • Example 2
  • the expression vectors obtained in example 1 were purified with the method of anion-exchange and exclusion chromatography.
  • the ready suspension comprised adenovirus particles in a buffer for the liquid form of the agent or in a buffer for lyophilized form of the agent.
  • the immunobiological agent based on the genome of recombinant human adenovirus serotype 26 in which El and E3 sites were deleted, while ORF6-Ad26 site was replaced with ORF6-Ad5 with the expression cassette comprising the CMV promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:l (Ad26- CMV-S-CoV2) in a buffer for the liquid form of the agent.
  • the immunobiological agent based on the genome of recombinant human adenovirus serotype 26 in which El and E3 sites were deleted, while ORF6-Ad26 site was replaced with ORF6-Ad5 with the expression cassette comprising the CMV promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:l (Ad26- CMV-S-CoV2) in a buffer for the lyophilized form of the agent.
  • the immunobiological agent based on the genome of recombinant human adenovirus serotype 26 in which El and E3 sites were deleted, while ORF6-Ad26 site was replaced with ORF6-Ad5 with the expression cassette comprising the CAG promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:2 (Ad26- CAG-S-CoV2) in a buffer for the liquid form of the agent.
  • the immunobiological agent based on the genome of recombinant human adenovirus serotype 26 in which El and E3 sites were deleted, while ORF6-Ad26 site was replaced with ORF6-Ad5 with the expression cassette comprising the CAG promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:2 (Ad26- CAG-S-CoV2) in a buffer for the lyophilized form of the agent.
  • the immunobiological agent based on the genome of recombinant human adenovirus serotype 26 in which El and E3 sites were deleted, while ORF6-Ad26 site was replaced with ORF6-Ad5 with the expression cassette comprising the EF1 promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:3 (Ad26- EFl-S-CoV2) in a buffer for the liquid form of the agent.
  • the immunobiological agent based on the genome of recombinant human adenovirus serotype 26 in which El and E3 sites were deleted, while ORF6-Ad26 site was replaced with ORF6-Ad5 with the expression cassette comprising the EF1 promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:3 (Ad26- EFl-S-CoV2) in a buffer for the lyophilized form of the agent.
  • Each of the given immunobiological agents is component 1 in variant 1 and in variant 2 of the developed agent.
  • the pSim25-Ends plasmid construct comprising two regions homologous to the genome of simian adenovirus serotype 25 (two homology arms).
  • One homology arm is the beginning of the genome of simian adenovirus serotype 25 (from the left inverted terminal repeat to El site) and the sequence from the end of El site to pIVa2 protein.
  • the second homology arm comprises the sequence of the end of adenovirus genome, including the right inverted terminal repeat.
  • the pSim25-Ends construct was synthesized by CJSC "Evrogen" (Moscow).
  • Simian adenovirus serotype 25 DNA isolated from virions was mixed with pSim25- Ends.
  • the pSim25-dlEl plasmid was obtained that comprised the genome of simian adenovirus serotype 25 with deleted El site.
  • the pSim25-dlEl plasmid E3 site of the adenovirus genome 3921 b.p. from the beginning of gene 12,5K to gene 14,7K was deleted to increase the vector packing capacity.
  • the pSim25-null plasmid construct was obtained that encoded the full genome of simian adenovirus serotype 25 with deletion of El and E3 sites.
  • the expression cassette SEQ ID NO:4 consists of the CMV promoter, gene of SARS-CoV-2 virus S protein and polyadenylation signal;
  • the expression cassette SEQ ID NO:2 consists of the CAG promoter, gene of SARS-CoV-2 virus S protein and polyadenylation signal;
  • the expression cassette SEQ ID NOG consists of the EF1 promoter, gene of SARS- CoV-2 virus S protein and polyadenylation signal.
  • constructs pArms-Sim25-CMV-S-CoV2, pArms-Sim25-CAG-S-CoV2, pArms-Sim25-EFl-S-CoV2 were obtained that comprised expression cassettes SEQ ID NO:4, SEQ ID NOG or SEQ ID NOG, respectively, as well as homology arms of simian adenovirus serotype 25.
  • the pSim25-CMV-S-CoV2, pSim25-CAG-S-CoV2, pSim25-EFl- S-CoV2 plasmids were hydrolyzed with the specific restriction endonuclease to delete the vector part.
  • the obtained DNA preparations were used to transfect HEK293 culture cells.
  • the obtained material was used to accumulate preparatory quantities of recombinant adenoviridae.
  • recombinant human adenoviridae serotype 25 comprising the gene of SARS-CoV-2 virus S protein: simAd25-CMV-S-CoV2 (comprising the expression cassette SEQ ID NO:4), simAd25-CAG-S-CoV2 (comprising the expression cassette SEQ ID NO:2), simAd25-EFl-S-CoV2 (comprising the expression cassette SEQ ID NO:3).
  • the expression vector was obtained that comprised the genome of recombinant simian adenovirus serotype 25 with deleted El and E3 sites with an integrated expression cassette selected from SEQ ID NO:4, SEQ ID NO:2, SEQ ID NO:3.
  • the expression vectors obtained in example 3 were purified with the method of anion-exchange and exclusion chromatography.
  • the ready suspension comprised adenovirus particles in a buffer for the liquid form of the agent or in a buffer for lyophilized form of the agent.
  • the immunobiological agent based on the genome of recombinant simian adenovirus serotype 25 with deleted El and E3 sites with the expression cassette comprising the CMV promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:4 (simAd25-CMV-S-CoV2) in a buffer for the liquid form of the agent.
  • the immunobiological agent based on the genome of recombinant simian adenovirus serotype 25 with deleted El and E3 sites with the expression cassette comprising the CMV promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:4 (Ad26-CMV-S-CoV2) in a buffer for the lyophilized form of the agent.
  • the immunobiological agent based on the genome of recombinant simian adenovirus serotype 25 with deleted El and E3 sites with the expression cassette comprising the CAG promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:4 (simAd25-CAG-S-CoV2) in a buffer for the liquid form of the agent.
  • the immunobiological agent based on the genome of recombinant simian adenovirus serotype 25 with deleted El and E3 sites with the expression cassette comprising the CAG promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:2 (simAd25-CAG-S-CoV2) in a buffer for the lyophilized form of the agent.
  • the immunobiological agent based on the genome of recombinant simian adenovirus serotype 25 with deleted El and E3 sites with the expression cassette comprising the EF1 promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NOG (simAd25-EFl-S-CoV2) in a buffer for the liquid form of the agent.
  • the immunobiological agent based on the genome of recombinant simian adenovirus serotype 25 with deleted El and E3 sites with the expression cassette comprising the EF1 promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NOG (simAd25-EFl-S-CoV2) in a buffer for the lyophilized form of the agent.
  • Each of the given immunobiological agents is component 2 in variant 1 of the developed agent and component 1 in variant 3 of the developed agent.
  • the design of the pAd5-Ends plasmid construct comprising two regions homologous to the genome of human adenovirus serotype 5 (two homology arms).
  • One homology arm is the beginning of the genome of human adenovirus serotype 5 (from the left inverted terminal repeat to El site) and the sequence of the virus genome comprising pIX protein.
  • the second homology arm comprises the nucleotide sequence after ORF3 E4 site to the genome end.
  • the pAd5-Ends construct was synthesized by CJSC "Evrogen" (Moscow). Human adenovirus serotype 5 DNA isolated from virions was mixed with pAd5- Ends. As a result of homologous recombination between pAd5-Ends and virus DNA the pAd5-dlEl plasmid was obtained that comprised the genome of human adenovirus serotype 5 with deleted El site.
  • the expression cassette SEQ ID NO:l consists of the CMV promoter, gene of SARS-CoV-2 virus S protein and polyadenylation signal;
  • the expression cassette SEQ ID NO:2 consists of the CAG promoter, gene of SARS-CoV-2 virus S protein and polyadenylation signal;
  • the expression cassette SEQ ID NO:3 consists of the EF1 promoter, gene of SARS- CoV-2 virus S protein and polyadenylation signal.
  • constructs pArms-Ad5-CMV-S-CoV2, pArms-Ad5-CAG-S-CoV2, pArms- Ad5-EFl-S-CoV2 were obtained that comprised expression cassettes SEQ ID NO:l, SEQ ID NO:2 or SEQ ID NOG, respectively, as well as homology arms of the genome of adenovirus serotype 5.
  • each plasmid was mixed with the recombinant vector pAd5-too-null.
  • the pAd5-too-CMV-S-CoV2, pAd5-too-GAC-S-CoV2, pAd5- too-EFl-S-CoV2 plasmids were hydrolyzed with the specific restriction endonuclease to delete the vector part.
  • the obtained DNA preparation was used to transfect HEK293 culture cells.
  • the obtained material was used to accumulate preparatory quantities of the recombinant adenovirus.
  • recombinant human adenoviridae serotype 5 comprising the gene of S SARS-CoV-2 virus S protein: Ad5-CMV-S-CoV2 (comprising the expression cassette SEQ ID NO:l), Ad5-CAG-S-CoV2 (comprising the expression cassette SEQ ID NO:2), Ad5-EFl-S-CoV2 (comprising the expression cassette SEQ ID NO:3).
  • the expression vector was obtained that comprised the genome of recombinant human adenovirus serotype 5 with deleted El and E3 sites with the integrated expression cassette selected from SEQ ID NO:l, SEQ ID NO:2, SEQ ID NO:3.
  • the expression vectors obtained in example 5 were purified with the method of anion-exchange and exclusion chromatography.
  • the ready suspension comprised adenovirus particles in a buffer for the liquid form of the agent or in a buffer for lyophilized form of the agent.
  • the immunobiological agent based on the genome of recombinant human adenovirus serotype 5 with deleted El and E3 sites with the expression cassette comprising the CMV promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:l (Ad5-CMV-S-CoV2) in a buffer for the liquid form of the agent.
  • the immunobiological agent based on the genome of recombinant human adenovirus serotype 5 with deleted El and E3 sites with the expression cassette comprising the CMV promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:l (Ad5-CMV-S-CoV2) in a buffer for the lyophilized form of the agent.
  • the immunobiological agent based on the genome of recombinant human adenovirus serotype 5 with deleted El and E3 sites with the expression cassette comprising the CAG promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:2 (Ad5-CAG-S-CoV2) in a buffer for the liquid form of the agent.
  • the immunobiological agent based on the genome of recombinant human adenovirus serotype 5 with deleted El and E3 sites with the expression cassette comprising the CAG promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NO:2 (Ad5-CAG-S-CoV2) in a buffer for the lyophilized form of the agent.
  • the immunobiological agent based on the genome of recombinant human adenovirus serotype 5 with deleted El and E3 sites with the expression cassette comprising the EF1 promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NOG (Ad5-EFl-S-CoV2) in a buffer for the liquid form of the agent.
  • the immunobiological agent based on the genome of recombinant human adenovirus serotype 5 with deleted El and E3 sites with the expression cassette comprising the EF1 promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, SEQ ID NOG (Ad5-EFl-S-CoV2) in a buffer for the lyophilized form of the agent.
  • Each of the given immunobiological agents is component 2 in variant 1 and in variant 3 of the developed agent.
  • Each component of the developed agent is an agent based on the recombinant adenovirus with an expression cassette in a buffer.
  • the authors of the invention developed a buffer composition that ensured stability of recombinant adenovirus particles.
  • the given solution comprises:
  • Tris(hydroxymethyl)aminomethane which is required to maintain pH of the solution.
  • EDTA which is used as a free-radical oxidation inhibitor.
  • Polysorbate-80 which is used as a surfactant.
  • the immunobiological agent based on the recombinant human adenovirus serotype 26 with the expression cassette comprising the CMV promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, l* 10 10 virus particles.
  • the immunobiological agent based on the recombinant human adenovirus serotype 5 with the expression cassette comprising the CMV promoter, the gene of SARS- CoV-2 virus S protein and polyadenylation signal, l* 10 10 virus particles.
  • stability of each of the serotypes of adenoviridae included in the agent was tested.
  • the obtained agents were stored at the temperature of -18°C and -70°C during 3 months, then defrosted and the change of the titer of the recombinant adenoviridae was analyzed.
  • the developed buffer for the liquid form of the agent ensures stability of all components of the developed agent in the following range of active ingredients (wt. %):
  • Tris 0.1831 wt. % ... 0.3432 wt. %;
  • Polysorbate-80 0.0378 wt. % ... 0.0709 wt. %;
  • Ethanol 95% 0.0004 wt. % ... 0.0007 wt. %;
  • the immunobiological agent based on the recombinant human adenovirus serotype 26 with the expression cassette comprising the CMV promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, 1*10 10 virus particles.
  • the immunobiological agent based on the recombinant human adenovirus serotype 5 with the expression cassette comprising the CMV promoter, the gene of SARS- CoV-2 virus S protein and polyadenylation signal, 1*10 10 virus particles.
  • the immunobiological agent based on the recombinant simian adenovirus serotype 25 with the expression cassette comprising the CMV promoter, the gene of SARS-CoV-2 virus S protein and polyadenylation signal, 1*10 10 virus particles.
  • the developed buffer for the lyophilized form of the agent ensures stability of all components of the developed agent in the following range of active ingredients:
  • Tris 0.0180 wt. % ... 0.0338 wt. %;
  • Magnesium chloride hexahydrate 0.0015 wt. % ... 0.0028 wt. %;
  • EDTA 0.0003 wt. % ... 0.0005 wt. %;
  • Polysorbate-80 0.0037 wt. % ... 0.0070 wt. %;
  • mice of BALB/c line were used.
  • the animals were distributed by groups of 10 and the following agents were administered to them: 1) Ad26-CMV-S-CoV2, intramuscularly (i/m), dose 5xl0 9 v.p./100 ul; in 21 days Ad26-CMV-S-CoV2, intramuscularly (i/m), dose 5x10 9 v.p./100 ul.
  • Ad26-CMV-S-CoV2 intranasally (i/n), dose 5xl0 9 v.p./100ul; in 21 days Ad26-CMV-S-CoV2 i/n, dose 5xl0 9 v.p./100ul.
  • the antigen (recombinant RBD) was adsorbed on the wells of a 96-well plate for ELISA during 16 hours at +4°C. 2) The, to get rid of non-specific binding, 5 % milk in TPBS 100 ul/well was added to the plate wells. It was incubated on a shaker at 37°C for an hour.
  • TMB tetramethylbenzidine
  • the titer of antibodies was determined as the latest dilution in which the optical density of the solution was reliably higher than in the negative control group.
  • the obtained results are given in Table 3.
  • an immunobiological agent was developed that is capable of inducing mucosal immune response against SARS-CoV-2 virus on the mucous membrane of the respiratory tract in children as well as a pattern of administration of this agent leading to potentiation of the mucosal immune response.
  • mice of BALB/c of different age were used. The animals were distributed into groups of 5:
  • the antigen (recombinant RBD) was adsorbed on the wells of a 96-well plate for ELISA during 16 hours at +4°C.
  • TMB tetramethylbenzidine
  • the titer of antibodies was determined as the latest dilution in which the optical density of the solution was reliably higher than in the negative control group.
  • the obtained results are given in Table 4.
  • the developed immunobiological agent induces development of humoral immune response both in adult and young animals of different age. This makes it possible to suggest that the agent will be effective in utilization for people of different age categories.
  • mice (aged 21-28 days) were used for the immunogenicity study. The animals were distributed into groups of 5 to whom the following agents were administered once: 1. Ad26-CMV-S-CoV2, intramuscularly (i/m), dose 10 10 vp/100 ul;
  • Cell immunity level was determined by assessing the quantity of proliferating CD4+ and CD8+ lymphocytes isolated from mouse spleen in the culture in vitro after repeated cell restimulation with recombinant S protein of SARS-CoV-2. Before immunization as well as after 14 days animals' spleens were harvested from which mononuclear cells were isolated by centrifugation in the ficoll solution density gradient (1.09 g/mL; PanEco). Then, the isolated cells were colored with the fluorescent dye CFSE (Invivogen, USA) and seeded on wells of a 96-well plate (2*10 5 cells/well). The repeated stimulation of lymphocytes was carried out in in vitro conditions by adding coronavirus S protein to the culture medium (final protein concentration 1 pg/ml). Intact cells to which the antigen was not added were used as the negative control.
  • CFSE fluorescent dye
  • proliferating cells were stained with antibodies to marker molecules of T lymphocytes CD3, CD4, CD8 (BDBioscienses, USA).
  • Proliferating (with a lower quantity of cell colorant CFSE) CD4+ and CD8+ T lymphocytes were determined in the cell mixture with the use of the flow cytofluorometer BD FACS Arialll (BD Biosciences, USA).
  • the resultant percentage of proliferating cells in each sample was determined by deducing the result obtained during analysis of intact cells from the result obtained during analysis of cells restimulated with coronavirus S protein antigen.
  • mice with the developed agent makes it possible to reliably increase the percentage of proliferating CD4+ and CD8+ T lymphocytes after antigen restimulation on Day 14 after immunization.
  • the antigen was adsorbed on the wells of a 96-well plate for ELISA during 16 hours at +4°C.
  • the serum samples of immunized mice were diluted by the method of 2-fold dilutions. In total, 12 dilutions of each sample were prepared.
  • TMB tetramethylbenzidine
  • the titer of IgG antibodies was determined as the latest dilution in which the optical density of the solution was reliably higher than in the negative control group. The obtained results (mean geometrical value) are given in Table 5.
  • Table 5- Titer Of Igg Antibodies To S Protein In Blood Serum Of Mice (Mean Geometrical Value Of The Titer Of Antibodies)
  • the aim of the given study was assessment of humoral immune response to S protein of SARS-CoV-2 with administration of the developed agent in different doses to young animals.
  • mice of C57/B16 line (3-4 weeks) were used to study immunogenicity of the developed agent.
  • the animals were distributed by groups of 5 to whom the following agents were administered intramuscularly, twice, with an interval of 28 days:
  • Ad5-CMV-S-CoV2 5*10 10 vp/100 ul.
  • the animals' blood was drawn, blood serum was isolated and the titer of IgG antibodies to S protein of SARS-CoV-2 coronavirus was determined by the ELISA method. To do that:
  • the antigen was adsorbed on the wells of a 96-well plate for ELISA during 16 hours at +4°C.
  • the serum samples of immunized mice were diluted 100 times and then by the method of 2-fold dilutions. In total, 12 dilutions of each sample were prepared.
  • TMB tetramethylbenzidine
  • the titer of antibodies was determined as the latest dilution in which the optical density of the solution was reliably higher than in the negative control group. The obtained results (mean geometrical value) are given in Table 6.
  • the developed agent demonstrates immunogenicity within the whole range of selected doses.
  • mice of C57/B16 line (3-4 weeks) were used to study immunogenicity of the developed agent.
  • the animals were distributed by groups of 5 and the following agents were administered to them:
  • the antigen was adsorbed on the wells of a 96-well plate for ELISA during 16 hours at +4°C.
  • the serum samples of immunized mice were diluted by the method of 2-fold dilutions. In total, 12 dilutions of each sample were prepared.
  • TMB tetramethylbenzidine
  • the reaction was stopped in 15 minutes by adding sulfuric acid. Then, using a spectrophotometer, the optical density (OD) of the solution in each well was measured at the wavelength of 450 nm.
  • the titer of antibodies was determined as the latest dilution in which the optical density of the solution was reliably higher than in the negative control group.
  • the obtained results are given in Table 7.
  • the obtained results demonstrate that the developed agent provides for development of humoral immune response against SARS-CoV-2 in young animals with different administration protocols.
  • the suggested immunization protocol included sequential intramuscular administration of component 1 and component 2 of the pharmaceutical agent according to variant 1 (Ad26-CMV-S-CoV2, Ad5- CMV-S-CoV2).
  • One volunteer discontinued participation before administration of component 1 because of newly diagnosed hypertension, correspondingly, 99 volunteers started the therapy under study.
  • Component 2 was not administered to: 1 person - because of non-attendance, 1 person - because of an adverse event (leukopenia), 1 person - because of an intestinal infection of unknown etiology (of enteroviral type), 1 person - because of hospitalization with a purulent furunculus in 5 days after administration of the component, 1 person - because of hospitalization (an intestinal infection) and 1 person - because of COVID, 2 people - refused to participate.
  • both vaccine components were received by 91 volunteers.
  • Table 8 includes the number (share) of volunteers with presence of AE in each group by the system organ class (SOC) and preferred term (PT) according to the MedDRA dictionary, as well as by connection with the preparation under study and severity degree.
  • SOC system organ class
  • PT preferred term
  • AEs were connected with the preparation under study (as the connection assessment is "possible”, “probable” or “certain”): tenderness at place of injection, influenza like syndrome, fatigue, subfebrile temperature, hot flash, nasal congestion arose immediately after vaccine injection, headache, sore throat, cough, joint ache, general weakness, stomachache, panic attack, reduction of neutrophils, increase of bilirubin level.
  • connection was determined as “doubtful” or "no connection". No allergic reactions to the preparation under study were observed.
  • the humoral immunity level was determined by assessing the titer of IgG antibodies specific to RBD domain of S protein of SARS-CoV-2 virus.
  • the participants of the study were divided into two groups:
  • component 1 and component 2 of the pharmaceutical agent 1) sequential intramuscular administration of component 1 and component 2 of the pharmaceutical agent by variant 1 (Ad26-CMV-S-CoV2, Ad5-CMV-S-CoV2) in the dose of lxl 0 10 virus particles/1 ml, 47 people.
  • variant 1 Ad26-CMV-S-CoV2, Ad5-CMV-S-CoV2
  • component 1 and component 2 of the pharmaceutical agent sequential intramuscular administration of component 1 and component 2 of the pharmaceutical agent by variant 1 (Ad26-CMV-S-CoV2, Ad5-CMV-S-CoV2) in the dose of 2x10 10 virus particles/1 ml, 48 people.
  • variant 1 Ad26-CMV-S-CoV2, Ad5-CMV-S-CoV2
  • the titer of antigen-specific IgG in the first group was assessed on Days 21, 28 and 42 of the study and in the second group - on Days 21 and 28 of the study.
  • the titer of antibodies was determined by means of a test system for ELISA that enables determining the titer of IgG to RBD domain of S antigen of SARS-CoV-2 virus.
  • the plates with pre-adsorbed RBD (100 ng/well) were washed 5 times with a washing buffer. Then, 100 ul of the positive control and 100 ul of the negative control were added to the plate wells in duplicate. A series of two-fold dilutions of the studied samples were added to other wells (two repeats per each sample). The plate was sealed with a film and incubated for an hour at +37 °C while mixing at the rate of 300 rpm. Then the wells were washed with a working solution of the washing buffer.
  • the titer of IgG was determined as maximum serum dilution at which OD450 serum value of an immunized participant exceeds the control serum value (serum of a participant before immunization) more than 2 times.
  • component 1 and component 2 of the pharmaceutical agent 1) sequential intramuscular administration of component 1 and component 2 of the pharmaceutical agent by variant 1 (Ad26-CMV-S-CoV2, Ad5-CMV-S-CoV2) in the dose of lxlO 10 virus particles/1 ml, 47 people.
  • variant 1 Ad26-CMV-S-CoV2, Ad5-CMV-S-CoV2
  • component 1 and component 2 of the pharmaceutical agent sequential intramuscular administration of component 1 and component 2 of the pharmaceutical agent by variant 1 (Ad26-CMV-S-CoV2, Ad5-CMV-S-CoV2) in the dose of 2xl0 10 virus particles/1 ml, 48 people.
  • variant 1 Ad26-CMV-S-CoV2, Ad5-CMV-S-CoV2
  • Cell immunity level was determined by assessing the quantity of proliferating CD4+ and CD 8+ lymphocytes of volunteers' peripheral blood in in vitro culture after repeated cell restimulation with recombinant S protein of SARS-CoV-2. Before immunization as well as after 28 days the patients' blood samples were drawn from which mononuclear cells were isolated by centrifugation in the ficoll solution density gradient (1.077 g/mL; PanEco). Then, the isolated cells were colored with the fluorescent dye CFSE (Invivogen, USA) and seeded on wells of a 96-well plate (2*10 5 cells/well).
  • CFSE fluorescent dye
  • the repeated stimulation of lymphocytes was carried out in in vitro conditions by adding coronavirus S protein to the culture medium (final protein concentration 1 pg/ml). Intact cells to which the antigen was not added were used as the negative control. In 72 hours after antigen addition % of proliferating cells was measured and the cultural medium was taken for measuring gamma interferon quantity.
  • proliferating cells were stained with antibodies to marker molecules of T lymphocytes CD3, CD4, CD8 (anti-CD3 Pe-Cy7 (BDBiosciences, SK7 clone), anti-CD4 APC (BDBiosciences, SK3 clone), anti-CD8 PerCP-Cy5.5 (BDBiosciences, SKI clone)).
  • Proliferating (with a lower quantity of cell colorant CFSE) CD4+ and CD8+ T lymphocytes were determined in the cell mixture with the use of the flow cytofluorometer BD FACS Arialll (BD Biosciences, USA). The resultant percentage of proliferating cells in each sample was determined by deducing the result obtained during analysis of intact cells from the result obtained during analysis of cells restimulated with coronavirus S protein antigen.
  • the provided examples confirm that as a result of conducted work a safe and effective agent was created that induced development of reactions of humoral and cell immune response against SARS-CoV-2 virus in children aged 1 month and older.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Virology (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Veterinary Medicine (AREA)
  • Biochemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Physics & Mathematics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Epidemiology (AREA)
  • Plant Pathology (AREA)
  • Mycology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Pulmonology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

Le groupe de l'invention concerne la biotechnologie, l'immunologie et la virologie. L'utilisation d'un agent contenant un vecteur d'expression basé sur une souche d'adénovirus humain sérotype 26 ou d'adénovirus humain sérotype 5 est décrite, dans laquelle les régions E1 et E3 sont délétées, avec une cassette d'expression intégrée choisie parmi les SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, ou d'adénovirus simien sérotype 25, dans lequel les régions E1 et E3 sont délétées avec une cassette d'expression intégrée choisie parmi les SEQ ID NO: 4, SEQ ID NO : 2, SEQ ID NO : 3 ou ne contient qu'un composant 2 pour l'induction d'une immunité spécifique contre le virus du syndrome respiratoire aigu sévère SARS-CoV-2 chez les enfants âgés de 1 mois et plus.
PCT/RU2022/000047 2021-11-26 2022-02-18 Induction d'une immunité contre le sras-cov-2 chez les enfants WO2022197209A1 (fr)

Priority Applications (8)

Application Number Priority Date Filing Date Title
CN202280000642.4A CN116528895A (zh) 2021-11-26 2022-02-18 用于在儿童中诱导针对sars-cov-2的特异性免疫的药剂的用途
EP22713843.5A EP4436599A1 (fr) 2021-11-26 2022-02-18 Induction d'une immunité contre le sras-cov-2 chez les enfants
KR1020227010857A KR20240135414A (ko) 2021-11-26 2022-02-18 아동의 중증 급성 호흡기 증후군 바이러스 SARS-COV-2에 대한 특이 면역(specific immunity) 유도제의 용도
CA3156264A CA3156264A1 (fr) 2021-11-26 2022-02-18 Utilisation de l'agent pour l'induction de l'immunite specifique contre le virus du syndrome respiratoire aigu severe (sras-cov-2) chez les enfants
JP2022520203A JP2023512381A (ja) 2021-11-26 2022-02-18 小児において重症急性呼吸器症候群ウイルスｓａｒｓ－ｃｏｖ－２に対する特異的免疫を誘導するための薬剤の利用
MX2022004059A MX2022004059A (es) 2021-11-26 2022-02-18 Utilizacion de un agente para la induccion de inmunidad especifica contra el virus del sindrome respiratorio agudo severo sars-cov-2 en ni?os.
ZA2022/03566A ZA202203566B (en) 2021-11-26 2022-03-28 Utilization of an agent for induction of specific immunity against severe acute respiratory syndrome virus sars-cov-2 in children
IL291816A IL291816B2 (en) 2021-11-26 2022-03-30 Drug administration to activate specific immunity against the severe acute respiratory syndrome virus SARS-COV-2 in children

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
RU2021134724 2021-11-26
RU2021134724A RU2761904C1 (ru) 2021-11-26 2021-11-26 Применение средства для индукции специфического иммунитета против вируса тяжелого острого респираторного синдрома SARS-CoV-2 у детей

Publications (1)

Publication Number Publication Date
WO2022197209A1 true WO2022197209A1 (fr) 2022-09-22

Family

ID=79175094

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/RU2022/000047 WO2022197209A1 (fr) 2021-11-26 2022-02-18 Induction d'une immunité contre le sras-cov-2 chez les enfants

Country Status (3)

Country Link
AR (1) AR124746A1 (fr)
RU (1) RU2761904C1 (fr)
WO (1) WO2022197209A1 (fr)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2720614C9 (ru) * 2020-04-23 2021-02-09 федеральное государственное бюджетное учреждение «Национальный исследовательский центр эпидемиологии и микробиологии имени почетного академика Н.Ф. Гамалеи» Министерства здравоохранения Российской Федерации Иммунобиологическое средство и способ его использования для индукции специфического иммунитета против вируса тяжелого острого респираторного синдрома SARS-CoV-2 (варианты)
RU2743963C1 (ru) * 2021-02-09 2021-03-01 федеральное государственное бюджетное учреждение «Национальный исследовательский центр эпидемиологии и микробиологии имени почетного академика Н.Ф. Гамалеи» Министерства здравоохранения Российской Федерации Средство для индукции специфического иммунитета против вируса тяжелого острого респираторного синдрома SARS-CoV-2 в жидкой форме (варианты)
RU2731342C9 (ru) * 2020-08-22 2021-10-05 федеральное государственное бюджетное учреждение "Национальный исследовательский центр эпидемиологии и микробиологии имени почетного академика Н.Ф. Гамалеи" Министерства здравоохранения Российской Федерации Фармацевтическое средство и способ его использования для индукции специфического иммунитета против вируса тяжелого острого респираторного синдрома SARS-CoV-2 (варианты)
WO2021204179A1 (fr) * 2020-04-09 2021-10-14 Suzhou Abogen Biosciences Co., Ltd. Vaccins à base d'acide nucléique pour coronavirus

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021205017A1 (fr) * 2020-04-09 2021-10-14 Valneva Austria Gmbh Améliorations apportées à des formulations de vaccin à usage médical

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021204179A1 (fr) * 2020-04-09 2021-10-14 Suzhou Abogen Biosciences Co., Ltd. Vaccins à base d'acide nucléique pour coronavirus
RU2720614C9 (ru) * 2020-04-23 2021-02-09 федеральное государственное бюджетное учреждение «Национальный исследовательский центр эпидемиологии и микробиологии имени почетного академика Н.Ф. Гамалеи» Министерства здравоохранения Российской Федерации Иммунобиологическое средство и способ его использования для индукции специфического иммунитета против вируса тяжелого острого респираторного синдрома SARS-CoV-2 (варианты)
RU2731342C9 (ru) * 2020-08-22 2021-10-05 федеральное государственное бюджетное учреждение "Национальный исследовательский центр эпидемиологии и микробиологии имени почетного академика Н.Ф. Гамалеи" Министерства здравоохранения Российской Федерации Фармацевтическое средство и способ его использования для индукции специфического иммунитета против вируса тяжелого острого респираторного синдрома SARS-CoV-2 (варианты)
RU2743963C1 (ru) * 2021-02-09 2021-03-01 федеральное государственное бюджетное учреждение «Национальный исследовательский центр эпидемиологии и микробиологии имени почетного академика Н.Ф. Гамалеи» Министерства здравоохранения Российской Федерации Средство для индукции специфического иммунитета против вируса тяжелого острого респираторного синдрома SARS-CoV-2 в жидкой форме (варианты)

Non-Patent Citations (15)

* Cited by examiner, † Cited by third party
Title
"GenBank", Database accession no. YP 009724390
A.K. SIMONG.A. HOLLANDERA. MCMICHAEL: "Evolution of the immune system in humans from infancy to old age", PROC BIOL SCI, vol. 282, no. 1821, 22 December 2015 (2015-12-22), pages 20143085
F.P. POLACKETAL.: "Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine", N ENGL J MED, vol. 383, 2020, pages 2603 - 2615, XP055820495, Retrieved from the Internet <URL:www.cdc.gov/coronavirus/2019-ncov/vaccines/recommendations/adolescents.html> DOI: 10.1056/NEJMoa2034577
FENG-CAI ZHU ET AL.: "Immunogenicity and safety of a recombinant adenovirus type-5-vectored COVID-19 vaccine in healthy adults aged 18 years or older: a randomised, double-blind, placebo-controlled, phase 2 trial", THE LANCET, vol. 369, 2020, pages 479 - 488, XP086249198, DOI: 10.1016/S0140-6736(20)31605-6
J IMMUNOL., vol. 176, no. 1, 1 January 2006 (2006-01-01), pages 165 - 72
J. SADOFF: "Interim Results of a Phase 1-2a Trial of Ad26.COV2.S Covid-19 Vaccine.", ENGL J MED, 13 January 2021 (2021-01-13)
JERALD SADOFF, LE GARS MATHIEU, SHUKAREV GEORGI, HEERWEGH DIRK, TRUYERS CARLA, DE GROOT ANNE M., STOOP JEROEN, TETE SARAH, VAN DAM: "Interim Results of a Phase 1–2a Trial of Ad26.COV2.S Covid-19 Vaccine", THE NEW ENGLAND JOURNAL OF MEDICINE, MASSACHUSETTS MEDICAL SOCIETY, US, 13 January 2021 (2021-01-13), US , XP055764899, ISSN: 0028-4793, DOI: 10.1056/NEJMoa2034201 *
KAUR KCHOWDHURY SGREENSPAN NSSCHREIBER JR: "Decreased expression of tumor necrosis factor family receptors involved in humoral immune responses in preterm neonates", BLOOD, vol. 110, no. 8, 15 October 2007 (2007-10-15), pages 2948 - 54
L. A. JACKSON ET AL.: "An mRNA Vaccine against SARS-CoV-2 — Preliminary Report", N ENGL J MED, vol. 383, 2020, pages 1920 - 1931, Retrieved from the Internet <URL:https://penntoday.upenn.edu/news/covid-vaccine-kids>
LIANG M ET AL.: "SARS patients-derived human recombinant antibodies to S and M proteins efficiently neutralize SARS-coronavirus infectivity", BIOMEDENVIRONSCI, vol. 18, no. 6, December 2005 (2005-12-01), pages 363 - 74, XP008139125
LISA A. JACKSON, ANDERSON EVAN J., ROUPHAEL NADINE G., ROBERTS PAUL C., MAKHENE MAMODIKOE, COLER RHEA N., MCCULLOUGH MICHELE P., C: "An mRNA Vaccine against SARS-CoV-2 — Preliminary Report", THE NEW ENGLAND JOURNAL OF MEDICINE, MASSACHUSETTS MEDICAL SOCIETY, US, 14 July 2020 (2020-07-14), US , XP055715132, ISSN: 0028-4793, DOI: 10.1056/NEJMoa2022483 *
M. VOYSEY ET AL.: "Safety and efficacy of the ChAdOxl nCoV-19 vaccine (AZD1222) against SARS-CoV-2: an interim analysis of four randomised controlled trials in Brazil, South Africa, and the UK", THELANCET, vol. 397, 2021, pages 99 - 111, XP086439265, DOI: 10.1016/S0140-6736(20)32661-1
SHCHEPLYAGINA, L.A.KRUGLOVA, I.V.: "Age peculiarities of immunity in children", RUSSIAN MEDICAL JOURNAL, no. 23, 11 November 2009 (2009-11-11), pages 1564
WANG W.JIA YL.LI YC.JING CQ.GUO X.SHANG XF.ZHAO CP.WANG TY: "Impact of different promoters, promoter mutation, and an enhancer on recombinant protein expression in CHO cells.", SCIENTIFIC REPORTS, vol. 8, 2017, pages 10416
YANG C.Q.LI X.Y.LI Q.FU S.L.LI H.GUO Z.K.LIN J.T.ZHAO S.T.: "Evaluation of three different promoters driving gene expression in developing chicken embryo by using in vivo electroporation", GENET. MOL. RES., vol. 13, 2014, pages 1270 - 1277

Also Published As

Publication number Publication date
RU2761904C1 (ru) 2021-12-13
AR124746A1 (es) 2023-05-03

Similar Documents

Publication Publication Date Title
RU2720614C1 (ru) Иммунобиологическое средство и способ его использования для индукции специфического иммунитета против вируса тяжелого острого респираторного синдрома SARS-CoV-2 (варианты)
WO2021254327A1 (fr) Vaccin à vecteur viral de type à remplacement d&#39;enveloppe et procédé de construction associé
Sui et al. Protection against SARS-CoV-2 infection by a mucosal vaccine in rhesus macaques
WO2021076010A1 (fr) Agent pharmaceutique pour induire une immunité spécifique contre le sras-cov-2
WO2018215766A1 (fr) Compositions et procédés d&#39;induction d&#39;une réponse immunitaire
RU2743963C1 (ru) Средство для индукции специфического иммунитета против вируса тяжелого острого респираторного синдрома SARS-CoV-2 в жидкой форме (варианты)
Cao et al. A single vaccine protects against SARS-CoV-2 and influenza virus in mice
JP7369276B2 (ja) 重症急性呼吸器症候群ウイルスｓａｒｓ－ｃｏｖ－２に対する発現ベクター
WO2022197209A1 (fr) Induction d&#39;une immunité contre le sras-cov-2 chez les enfants
EP4436599A1 (fr) Induction d&#39;une immunité contre le sras-cov-2 chez les enfants
RU2709659C1 (ru) Иммунобиологическое средство и способ его использования для индукции специфического иммунитета к вирусу ближневосточного респираторного синдрома (варианты)
JP2023501868A (ja) 重症急性呼吸器症候群ウイルスＳＡＲＳ－ＣｏＶ－２に対する特異的免疫を誘導するための凍結乾燥形態（変異型）の薬剤
US20220265816A1 (en) Use of the agent for induction of specific immunity against severe acute respiratory syndrome virus sars-cov-2 for revaccination of population (variants)
JP7360544B2 (ja) Ｓａｒｓ－ｃｏｖ－２に対する特異的免疫を誘導するための医薬品
EA043182B1 (ru) Применение средства для индукции специфического иммунитета против вируса тяжелого острого респираторного синдрома sars-cov-2 у детей
WO2022177466A1 (fr) Utilisation d&#39;un agent pour induire une immunité contre le sars-cov-2
JP2023507543A (ja) 年齢６０歳超であり、そして／あるいは慢性疾患を有する対象において重症急性呼吸器症候群ウイルスＳＡＲＳ－ＣｏＶ－２に対する特異的免疫を誘導するための作用物質（変形例）の使用
EA042672B1 (ru) ПРИМЕНЕНИЕ СРЕДСТВА ДЛЯ ИНДУКЦИИ СПЕЦИФИЧЕСКОГО ИММУНИТЕТА ПРОТИВ ВИРУСА ТЯЖЕЛОГО ОСТРОГО РЕСПИРАТОРНОГО СИНДРОМА SARS-CoV-2 У ЛИЦ СТАРШЕ 60 ЛЕТ И/ИЛИ ИМЕЮЩИХ ХРОНИЧЕСКИЕ ЗАБОЛЕВАНИЯ (ВАРИАНТЫ)
EA043163B1 (ru) Применение средства для индукции специфического иммунитета против вируса тяжелого острого респираторного синдрома sars-cov-2 для ревакцинации населения (варианты)
WO2022177465A1 (fr) Utilisation d&#39;un agent pour induire une immunité contre le sars-cov-2

Legal Events

Date Code Title Description
ENP Entry into the national phase

Ref document number: 2022520203

Country of ref document: JP

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 202280000642.4

Country of ref document: CN

WWE Wipo information: entry into national phase

Ref document number: 202227020559

Country of ref document: IN

WWE Wipo information: entry into national phase

Ref document number: 140150140003000278

Country of ref document: IR

ENP Entry into the national phase

Ref document number: 2022713843

Country of ref document: EP

Effective date: 20220408

REG Reference to national code

Ref country code: BR

Ref legal event code: B01A

Ref document number: 112022005904

Country of ref document: BR

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

Ref document number: 22713843

Country of ref document: EP

Kind code of ref document: A1

REG Reference to national code

Ref country code: BR

Ref legal event code: B01E

Ref document number: 112022005904

Country of ref document: BR

Free format text: APRESENTAR DOCUMENTOS COMPROBATORIOS QUE EXPLIQUEM E REGULARIZEM A DIVERGENCIA NO NOME DOS INVENTORES CONSTANTES NA PUBLICACAO INTERNACIONAL ELIZAVETA ALEXANDROVNA TOKARSKAYA, ALINA SERGEEVNA EROKHOVA, NATALIA ANATOLIEVNA NIKITENKO, SVETLANA YAKOVLEVNA LOGINOVA E DMITRII ANATOLIEVICH KUTAEV E OS CONSTANTES NO FORMULARIO DA PETICAO INICIAL ELIZAVETA ALEXSANDROVNA TOKARSKAYA, ALINA SERGEEVNA EROXOVA, NATALIA ANATOLEVNA NIKITENKO, SVETLANA IAKOVLEVNA LOGINOVA E DMITRII ANATOLEVICH KUTAEV UMA VEZ QUE NAO HOUVE ENVIO DE DOCUMENTO COMPROVANDO QUE O NOME CORRETO E O DECLARADO NA ENTRADA NACIONAL.

ENP Entry into the national phase

Ref document number: 112022005904

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20220328

NENP Non-entry into the national phase

Ref country code: DE