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KR-102963944-B1 - coronavirus vaccine

KR102963944B1KR 102963944 B1KR102963944 B1KR 102963944B1KR-102963944-B1

Abstract

The present invention relates to nucleic acids suitable for the treatment or prevention of coronavirus, preferably coronavirus SARS-CoV-2 infection, or disorders associated with such infection, preferably COVID-19. The present invention also relates to compositions, polypeptides, and vaccines. The compositions and vaccines preferably comprise at least one nucleic acid sequence, preferably a nucleic acid sequence associated with lipid nanoparticles (LNPs). The present invention also relates to first and second medical uses of the nucleic acids, compositions, polypeptides, combinations, vaccines, and kits, and methods for treating or preventing coronavirus infection, preferably Coronavirus infection.

Inventors

  • 라우치, 수잔
  • 그로쎄, 한스 볼프강
  • 페취, 벤자민

Assignees

  • 큐어백 에스이

Dates

Publication Date
20260511
Application Date
20210203
Priority Date
20200204

Claims (20)

  1. Purified RNA containing the following: (a) 5' cap structure; (b) a protein derived from the SARS-CoV-2 coronavirus spike protein, at least one coding sequence encoding a pre-fusion stabilized spike protein (S_stab) which is at least 90% identical to SEQ ID NO. 10, includes K986P and V987P stabilizing substitutions compared to SEQ ID NO. 10, and further includes a D614G amino acid substitution; and (c) At least one heterologous 3' untranslated region (UTR) comprising at least one poly(A) sequence having 30 to 200 adenosine nucleotides.
  2. Purified RNA according to claim 1, wherein the 5' cap is an m7G, cap0, cap1, cap2, modified cap0, or modified cap1 structure.
  3. In paragraph 2, purified RNA having a 5' cap of the cap1 structure.
  4. The purified RNA of claim 1, wherein the purified RNA comprises a terminal poly(A) sequence of 30 to 200 adenosine nucleotides.
  5. Purified RNA according to claim 1, further comprising at least one heterologous 5' UTR.
  6. In claim 1, at least one coding sequence encoding the SARS-CoV-2 spike protein is purified RNA having a G/C content of at least 50%.
  7. In paragraph 1, purified RNA is a replicon RNA.
  8. In paragraph 7, purified RNA in which the replicon RNA encodes a replicase element derived from an alphavirus.
  9. In paragraph 8, purified RNA in which the replicon RNA encodes a replicase element derived from VEE.
  10. Purified RNA according to claim 1, comprising a 1-methylpseudouridine nucleotide substitution at one or more uracil positions.
  11. In claim 1, the purified RNA is purified by RP-HPLC and/or TFF.
  12. In paragraph 11, purified RNA that is purified by TFF.
  13. In claim 1, purified RNA in which the protein derived from the SARS-CoV-2 coronavirus spike protein contains an amino acid sequence that is 95% identical to SEQ ID NO. 10.
  14. In claim 13, purified RNA in which at least one coding sequence comprises a nucleic acid sequence that is at least 80% identical to sequence number 137.
  15. In claim 14, purified RNA in which at least one coding sequence comprises a nucleic acid sequence that is at least 90% identical to sequence number 137.
  16. A pharmaceutical composition for use in the treatment or prevention of coronavirus infection, comprising purified RNA according to any one of claims 1 to 15 and at least one pharmaceutically acceptable carrier.
  17. A composition according to claim 16 in which RNA is complexed or associated with lipid nanoparticles (LNPs).
  18. In paragraph 17, a composition in which the LNPs comprise the following: (i) at least one cationic lipid component; (ii) at least one neutral lipid component; (iii) at least one sterol component; and (iv) At least one PEG-lipid component.
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Description

coronavirus vaccine The present invention relates particularly to nucleic acids suitable for use in the treatment or prevention of infections caused by coronaviruses, preferably coronavirus SARS-CoV-2, or disorders associated with such infections, preferably COVID-19. The present invention also relates to compositions, polypeptides, and vaccines. The compositions and vaccines preferably comprise at least one nucleic acid sequence, preferably a nucleic acid sequence associated with a polymer carrier, a polycationic protein or peptide, or a lipid nanoparticle (LNP). The present invention also relates to first and second medical uses of said nucleic acids, said compositions, said polypeptides, said vaccines, and said kits, and methods for treating or preventing coronavirus infections, preferably SARS-CoV-2 infections. Coronavirus is an enveloped, positive single-stranded RNA virus of the Coronaviridae family. They typically cause a wide variety of diseases in various vertebrates, such as mammals, birds, and fish. Coronaviruses are genetically highly variable, and individual virus species can overcome species barriers to infect multiple host species. This migration led to human infections with SARS-associated coronavirus (SARS-CoV) and Middle East Respiratory Syndrome coronavirus (MERS-CoV). The coronavirus epidemic that began in Wuhan, China, during the transition period of 2019/2020 is attributed to a previously unknown coronavirus given the preliminary name nCoV-2019 or Wuhan Human Coronavirus (WHCV). This virus was later officially named SARS-CoV-2. Typical symptoms of a viral infection caused by SARS-CoV-2, also known as COVID-19 (Coronavirus disease 2019), include fever, cough, shortness of breath, pneumonia, and gastrointestinal symptoms (e.g., diarrhea). Severe illness can lead to respiratory failure requiring mechanical ventilation and support in the intensive care unit. On January 30, 2020, the World Health Organization (WHO) declared a global health emergency regarding the novel coronavirus infection. On March 11, the WHO declared COVID-19 a pandemic, noting over 118,000 cases of coronavirus disease across more than 110 countries and territories worldwide and the continuing risk of further global spread. By the end of March 2020, over 800,000 cases of SARS-CoV-2 infection had been confirmed, the virus had spread to almost every country in the world, and more than 40,000 COVID-19-related deaths had occurred. Currently, there are no vaccines or specific treatments available for SARS-CoV-2 infection and/or COVID-19 disease. Patients diagnosed with SARS-CoV-2 infection receive only supportive care based on their individual symptoms and clinical condition. Due to the significant risk of a severe global pandemic, safe and effective treatment or prevention of SARS-CoV-2 infection is urgently needed. A vaccine is particularly necessary to protect the elderly, who have a high mortality rate. Nucleic acid-based vaccination, including DNA or RNA, represents a promising technology for new vaccines against novel viruses. Nucleic acids can be genetically engineered and administered to human subjects. Transfected cells directly produce encoded antigens (e.g., DNA or RNA, particularly provided by mRNA) to trigger a protective immunological response. The pivotal role of virus-specific memory T cells in extensive and long-term protection against SARS-CoV infection has been revealed (e.g., see Channappanavar, Rudragouda, et al. "Virus-specific memory CD8 T cells provide substantial protection from lethal severe acute respiratory syndrome coronavirus infection." Journal of virology 88.19 (2014): 11034-11044). Virus-specific CD8 T cells are required, for example, for pathogen clearance and for mediating protection after viral infection. Therefore, an effective SARS-CoV-2 vaccine not only induces a potent functional humoral immune response but also induces SARS-CoV-2-specific CD8+ T cell and CD4+ T cell responses. Figure 1 shows that mRNA constructs encoding different SARS-CoV-2 S protein designs induce detectable protein expression using the in vitro translation system. Specific details are provided in Example 2a and Table 5 . Figure 2 shows, using FACS analysis, that mRNA constructs encoding different SARS-CoV-2 S protein designs are expressed on the cell surface of mammalian cells. Specific details are provided in Example 2b and Table 6 . Figure 3 shows that mRNA constructs encoding different SARS-CoV-2 S proteins are expressed in mammalian cells using Western blot analysis. Specific details are provided in Example 2c and Table 7 . Figure 4 shows significant IgG1 and IgG2a responses for the group vaccinated with the mRNA vaccine encoding the full-length stabilized S protein. Figure 4 shows comparable IgG1 responses for the mRNA vaccine and the rec. SARS-CoV-S protein, as well as higher IgG2a titers for the mRNA vaccine compared to the rec. SARS-CoV-S protein. IgG1 and IgG2a antibody titers were evaluated by ELISA using