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US-12622959-B2 - Coronavirus recombinant spike protein, polynucleotide encoding same, vector comprising polynucleotide, and vaccine for preventing or treating coronavirus infection, comprising vector

US12622959B2US 12622959 B2US12622959 B2US 12622959B2US-12622959-B2

Abstract

The present invention relates to a novel coronavirus recombinant spike protein, a polynucleotide encoding the same, a vector comprising the polynucleotide, and a vaccine for preventing or treating coronavirus infection, comprising the vector. The coronavirus recombinant spike protein of the present invention is stable and thereby not easily decomposed in cells, and effectively activates immune cells thereby resulting in a high antibody production amount and T cell reactivity. It was confirmed that the vector of the present invention exhibits a high antigen expression level and thereby has a high antibody production amount and T cell reactivity, has a long antibody production period and expression period, and does not show liver toxicity. Accordingly, the vector of the present invention can be helpfully used as a vaccine for preventing or treating coronavirus infection.

Inventors

  • Chang-Yuil Kang
  • Seung-Phil SHIN
  • Kwang-Soo SHIN
  • Tae-Gwon OH

Assignees

  • CELLID CO., LTD.

Dates

Publication Date
20260512
Application Date
20210827
Priority Date
20200827

Claims (14)

  1. 1 . A vector expressing a recombinant spike protein derived from a coronavirus, wherein the vector comprises a polynucleotide encoding the recombinant spike protein in which an amino acid sequence TNSPRRAR (SEQ ID NO: 32) of a cleavage site between S1 and S2 genes of a coronavirus spike protein is replaced by a linker sequence comprising an amino acid sequence of TGGGGSR (SEQ ID NO: 19), wherein the vector is derived from an adenoviral vector comprising the sequence as set forth in SEQ ID NO: 21, wherein E1 and E3 regions of the adenoviral vector are deleted, and wherein the polynucleotide encoding the recombinant spike protein is inserted in the deleted E1 region.
  2. 2 . The vector according to claim 1 , wherein the coronavirus is SARS-COV, MERS-COV or SARS-COV-2.
  3. 3 . The vector according to claim 2 , wherein the coronavirus is SARS-COV-2.
  4. 4 . The vector according to claim 1 , wherein the spike protein linked by the linker sequence has an increased stability.
  5. 5 . The according to claim 1 , wherein the spike protein linked by the linker sequence has an increased antigen expression level.
  6. 6 . The vector according to claim 1 , wherein the coronavirus is SARS-COV, MERS-COV or SARS-COV-2.
  7. 7 . The vector according to claim 1 , wherein the coronavirus is SARS-COV-2.
  8. 8 . The vector according to claim 1 , wherein the vector is a virus or a plasmid.
  9. 9 . The vector according to claim 8 , wherein the virus is adenovirus, retrovirus, lentivirus, modified vaccinia virus ankara (MVA), adeno-associated virus (AAV), herpes simplex virus or baculovirus.
  10. 10 . The vector according to claim 9 , wherein the adenovirus is Ad2, Ad4, Ad5, Ad11, Ad26, Ad35, ChAd68, FAd9 or PAd3.
  11. 11 . The vector according to claim 9 , wherein the adenovirus has a high antigen expression level.
  12. 12 . The vector according to claim 10 , wherein the adenovirus has a high antigen expression level.
  13. 13 . A method for preventing or treating coronavirus infection comprising a step of administering to a subject a vector of claim 1 .
  14. 14 . The method for preventing or treating coronavirus infection according to claim 13 , wherein the coronavirus infection is SARS, MERS or COVID-19.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a U.S. National Stage Application filed under 35 U.S.C. § 371 of International Application No. PCT/KR2021/011512, filed Aug. 27, 2021, which claims the benefit of and priority to Korean Application No. 10-2020-0108276, filed Aug. 27, 2020, and Korean Application No. 10-2020-0152184, filed Nov. 13, 2020, all of which are hereby incorporated by reference in their entireties to the extent not inconsistent herewith. REFERENCE TO AN ELECTRONIC SEQUENCE LISTING The contents of the electronic sequence listing (“3-23 revised seq listing 2026 ST25. txt”; Size: 545,662 bytes; and Date of Creation: Feb. 6, 2026) is herein incorporated by reference in its entirety. BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel coronavirus recombinant spike protein, a polynucleotide encoding the same, a vector comprising the polynucleotide, and a vaccine for preventing or treating coronavirus infection comprising the vector. 2. Description of the Related Art Coronavirus, which causes severe respiratory disease and causes death, is classified as RNA virus belonging to Coronaviridae, and coronavirus infection is defined as respiratory syndrome caused by infection with coronavirus. Among the viruses belonging to Coronaviridae, there are a total of seven types of viruses known to infect humans, including four types that cause colds (229E, OC43, NL63 and HKU1), two types that cause severe pneumonia (SARS-CoV and MERS-CoV), and SARS-CoV-2, the virus responsible for this pandemic. The three types of viruses that cause severe pneumonia (SARS-CoV, MERS-CoV and SARS-CoV-2) are spreading worldwide, starting with SARS-CoV in 2002, followed by MERS-CoV in 2012, and the current SARS-CoV-2, which has high gene sequence homology with SARS-CoV. Coronaviruses (Coronaviridae) have unstable RNAs as their genomes, so mutations easily occur, and due to these characteristics, there is a possibility of transmission in both animals and humans. Studies to date have shown that SARS-CoV has been transmitted from civets to humans, and MERS-CoV has been transmitted from dromedaries to humans. Judging from these cases, coronaviruses of other species that use animals as hosts have the potential to evolve into mutant viruses that infect humans. SARS-CoV-2 was first reported in late 2019. Compared to SARS-CoV that occurred in China in 2002, SARS-CoV has a higher severity, but SARS-CoV-2 has a higher transmissibility, which is attributed to mutations in spike protein, the cell receptor binding site of SARS-CoV-2. This resulted in a worldwide pandemic. Common signs of infection include respiratory symptoms, fever, cough, shortness of breath, and dyspne. In more severe cases, infection can cause pneumonia, severe acute respiratory syndrome, kidney failure and even death. Currently, numerous countries and institutions are striving to develop vaccines to eradicate the COVID-19 pandemic. The types of preventive vaccines are classified into inactivated vaccines, attenuated vaccines, protein subunit vaccines, viral vector-based vaccines, DNA vaccines, and mRNA vaccines depending on the production method. Inactivated vaccines and attenuated live vaccines, which have traditionally been widely used as vaccine formulations, have the advantage of a simple production process, but the inactivated vaccines do not induce a high level of immune response, and the live attenuated vaccines have a risk of reacquiring pathogenicity through mutations. Since the protein subunit vaccines do not induce an immune response well, there is a problem that an immune enhancer must be used together. With the development of biotechnology, vaccine formulations based on new platforms have begun to be developed, one of which is a viral vector-based vaccine. Vector-based vaccines activate the immune response by delivering vaccine antigen genes to human cells with high efficiency to produce antigen proteins on their own in the body. Viral vector-based vaccines are safe and induce a high level of immune response, so efficient immunization is possible only with a single administration without the need for repeated administration. Viral vector-based vaccines also have the advantage of efficiently inducing cytotoxic T cell immunity as well as antibody production based on the virus structurally. Many viruses are being studied as gene vectors, and among them, adenovirus is widely used in the field of gene therapy because it is easy to manipulate and safety has been verified through a lot of research. Adenovirus has high competitiveness compared to other viral vectors because it can deliver relatively large antigens (9-35 kb). Adenovirus is a linear, double-stranded DNA virus, 70-90 nm in diameter, has no envelope, and has an icosahedral capsid, which is formed by 240 hexons, 12 pentons, and fibers extending from each vertex of the icosahedron. These hexons, pentons and fibers determine the major adenovirus an