APPLICATION OF NOVEL CORONAVIRUS VACCINE PEPTIDE AND NANOEMULSION PREPARATION THEREOF IN PREVENTION OF NOVEL CORONAVIRUS WILD AND MUTANT STRAINS

20250009871

18/564932

May 30, 2022

Disclosed are an application of a coronavirus SARS-CoV-2 vaccine polypeptide, a polypeptide composition and a nanoemulsion preparation thereof in the prevention of coronavirus SARS-CoV-2 wild and mutant strain infections. Specifically, provided is a coronavirus SARS-CoV-2 vaccine polypeptide having an amino acid sequence derived from an S protein of SARS-CoV-2 wild and mutant strains, the vaccine polypeptide can enable the body to generate high-level and durable humoral immune responses against SARS-CoV-2 and to produce high titers of RBD-binding antibodies and neutralizing antibodies that block the binding of RBD to ACE2. The vaccine polypeptide can be used to prevent infections of SARS-CoV-2 wild strain and B.1.1.7, B.1.351, B.1.617, B.1.1.529 and other mutant strains.

1. A vaccine polypeptide against coronavirus SARS-CoV-2, wherein the vaccine polypeptide has an amino acid sequence derived from the S protein of SARS-CoV-2 wild-type and mutant strains; and the vaccine polypeptide is selected from the group consisting of: (a) a polypeptide having the amino acid sequence shown in any one of SEQ ID NOs: 1-15; (b) a derivative polypeptide formed by one or more amino acids addition, one or more amino acids substitution, or 1-3 amino acids deletion to the amino acid sequence of the polypeptide in (a), and the derivative polypeptide has essentially identical function to the original polypeptide before derivatization.

2. The vaccine polypeptide of claim 1, wherein the vaccine polypeptide can induce primates to produce wild-type and mutant RBD specific binding antibodies and neutralizing antibodies that block the binding of wild-type and mutant RBD to ACE2.

3. The vaccine polypeptide of claim 1, wherein the SARS-CoV-2 mutant strains are selected from the group consisting of: D614G mutant strain, B.1.1.7 mutant strain, B.1.351 mutant strain, P.1 mutant strain, B.1.617 mutant strain, B.1.1.529 mutant strain, and a combination thereof.

4. The vaccine polypeptide of claim 1, wherein the structure of the vaccine polypeptide is as shown in Formula I: X1-X-X2  (I), wherein, (a) X is a core fragment, wherein the sequence of the core fragment is selected from one or more of SEQ ID NOs: 1-15 (see Table A); (b) X1 and X2 each independently represent none, 1, 2, or 3 amino acids, and the total number of amino acids of X1 and X2 is ≤4, preferably is 3, 2, or 1, and more preferably is 0 or 1; (c) “-” represents a peptide bond, peptide linker, or other linker (that is, X1 and X, and/or, X and X2 are connected by peptide bonds, peptide linkers (such as a flexible linker consisting of 1-15 amino acids) or other linkers).

5. The vaccine polypeptide of claim 1, wherein the vaccine polypeptide is selected from Table A: [table included]

6. The vaccine polypeptide of claim 4, wherein X1 or X2 each independently represents none, K, C, G, L, or A.

7. The vaccine polypeptide of claim 1, wherein the vaccine polypeptide has at least one T cell epitope and/or at least one B cell epitope of the SARS-CoV-2 S protein.

8. An isolated peptide set, wherein the peptide set comprises at least two vaccine polypeptides against coronavirus SARS-CoV-2 of claim 1.

9. The isolated peptide set of claim 8, wherein the peptide set comprises the vaccine polypeptide selected from the group consisting of: (Z1) 1 or 2 vaccine polypeptides selected from SEQ ID Nos: 1 and 12, and 1, 2, 3, or 4 vaccine polypeptides selected from SEQ ID Nos: 2-5; (Z2) 1 or 2 vaccine polypeptides selected from SEQ ID Nos: 6 and 9, 1 or 2 vaccine polypeptides selected from SEQ ID Nos: 7 and 10, and 1 or 2 vaccine polypeptides selected from SEQ ID Nos: 8 and 11; (Z3) one or more vaccine polypeptides selected from SEQ ID Nos: 1-12, and 1, 2 or 3 vaccine polypeptides selected from SEQ ID Nos: 13-15.

10. The isolated peptide set of claim 8, wherein the peptide set comprises the vaccine polypeptide selected from the group consisting of: (s1) the vaccine polypeptide shown in SEQ ID No: 1, and the vaccine polypeptide shown in SEQ ID No: 2; (s2) 1 or 2 vaccine polypeptides selected from SEQ ID Nos: 1 and 12, and 1, 2, 3 or 4 vaccine polypeptides selected from SEQ ID Nos: 2-5, and the peptide set at least comprises one or more vaccine polypeptides selected from SEQ ID Nos: 3, 4, 5, and 12; (s3) the vaccine polypeptides shown in SEQ ID Nos: 6, 7 and 8; (s4) the vaccine polypeptides shown in SEQ ID Nos: 9, 10 and 11; (s5) the vaccine polypeptides shown in SEQ ID Nos: 6, 10 and 11; (s6) the vaccine polypeptides shown in SEQ ID Nos: 9, 7 and 11; (s7) the vaccine polypeptides shown in SEQ ID Nos: 9, 10 and 8; (s8) the vaccine polypeptides shown in SEQ ID Nos: 2, 9, 10 and 11; (s9) the vaccine polypeptides shown in SEQ ID Nos: 2, 6, 7 and 8.

11. A pharmaceutical composition comprising: the vaccine polypeptide against coronavirus SARS-CoV-2 of claim 1 or a peptide set comprising at least two of the vaccine polypeptides against coronavirus SARS-CoV-2, and a pharmaceutically acceptable carrier.

12. A vaccine polypeptide nanoemulsion formulation against coronavirus SARS-CoV-2, which comprises: (a) the vaccine polypeptide against coronavirus SARS-CoV-2 of claim 1; (b) an adjuvant, preferably the adjuvant is an oil-in-water nanoemulsion based on squalene; and (c) a pharmaceutically acceptable carrier, excipient or diluent.

13. (canceled)

14. A cell preparation, which comprises (a) immune cells which are immune activated by the vaccine polypeptide against coronavirus SARS-CoV-2 of claim 1 or a peptide set comprising at least two of the vaccine polypeptides against coronavirus SARS-CoV-2; and (b) a pharmaceutically acceptable carrier.

15. A method for generating an immune response to the coronavirus SARS-CoV-2 wild-type and mutant strains, which comprises the step of: administering the vaccine polypeptide against coronavirus SARS-CoV-2 of claim 1, or a peptide set comprising at least two of the vaccine polypeptides against coronavirus SARS-CoV-2, to a subject in need thereof.

16. The vaccine polypeptide nanoemulsion formulation of claim 12, wherein the formulation comprises a nanoemulsion based on squalene and an emulsifier, and the squalene is derived from shark liver, and the emulsifier is phospholipid, or polysorbate 80, or a combination of polysorbate 80 and phospholipids, as well as one or more of sucrose esters, citric acid fatty acid glycerides, fatty acid glycerides, fatty acid sorbitans, cyclodextrins, polyoxyethylene fatty acid esters.

61; 61; 61; 07

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