Mutant of L1 protein of human papillomavirus type 58

Patent metadata

Field	Value
Publication number	US-10940194-B2
Application number	US-201615781035-A
Country	US
Kind code	B2
Filing date	Dec 2, 2016
Priority date	Dec 4, 2015
Publication date	Mar 9, 2021
Grant date	Mar 9, 2021

Abstract

Provided are a mutated HPV58 L1 protein or a variant thereof, a sequence encoding the same, a method for preparing the same, and a virus-like particle comprising the same, wherein the protein or a variant thereof and the virus-like particle can induce the generation of neutralizing antibodies against at least two HPV types, and therefore can be used to prevent infection by said at least two HPV types, and a disease caused by said infection, such as cervical cancer and condyloma acuminatum. The invention further relates to the use of the protein and the virus-like particle in the manufacture of a pharmaceutical composition or a vaccine for preventing infection by said at least two HPV types, and a disease caused by said infection, such as cervical cancer and condyloma acuminatum.

First claim

The invention claimed is:

1. A mutated HPV58 L1 protein, wherein as compared with a wild type HPV58 L1 protein, the mutated HPV58 L1 protein has the following mutations:
(1) N-terminal truncation of 5-70 amino acids; and
(2)(a) substitution of amino acid residues at positions 80-87 of the wild type HPV58 L1 protein with amino acid residues at the corresponding positions of the wild-type HPV33 L1 protein; or
(2)(b) substitution of amino acid residues at positions 376-383 of the wild type HPV58 L1 protein with amino acid residues at the corresponding positions of the wild-type HPV33 L1 protein;
wherein said wild type HPV58 L1 protein has an amino acid sequence as set forth in SEQ ID NO: 1 or a sequence having 95% or greater identity to SEQ ID NO: 1; and wherein said wild type HPV33 L1 protein has an amino acid sequence as set forth in SEQ ID NO: 2 or a sequence having 95% or greater identity to SEQ ID NO: 2.

2. An isolated nucleic acid, encoding the mutated HPV58 L1 protein according to claim 1 .

3. A vector comprising the isolated nucleic acid according to claim 2 .

4. An isolated host cell comprising the isolated nucleic acid according to claim 2 and/or a vector comprising the isolated nucleic acid.

5. A HPV virus-like particle, comprising or consisting of the mutated HPV58 L1 protein according to claim 1 .

6. A pharmaceutical composition or vaccine, comprising the HPV virus-like particle according to claim 5 , and a pharmaceutically acceptable carrier and/or excipient.

7. A method for preparing the mutated HPV58 L1 protein according to claim 1 , comprising expressing the mutated HPV58 L1 protein in a host cell, and then recovering the mutated HPV58 L1 protein from a culture of the host cell.

8. A method for preparing a vaccine, comprising combining the HPV virus-like particle according to claim 5 with a pharmaceutically acceptable carrier and/or excipient.

9. A method for preventing HPV infection or a disease caused by HPV infection, comprising administering to a subject a prophylactically effective amount of the HPV virus-like particle according to claim 5 or a pharmaceutical composition or vaccine comprising the HPV virus-like particle and a pharmaceutically acceptable carrier and/or excipient.

10. The mutated HPV58 L1 protein according to claim 1 , wherein the mutated HPV58 L1 protein is characterized by one or more of the following items:
(i) the mutated HPV58 L1 protein has 5, 15, 27, 35, 40, 60 or 70 amino acids truncated at N-terminal, as compared with the wild type HPV58 L1 protein;
(ii) the amino acid residues at the corresponding positions as described in claim 1 section (2)(a) are amino acid residues at positions 54-61 of the wild type HPV33 L1 protein; and
(iii) the amino acid residues at the corresponding positions as described in claim 1 section (2)(b) are amino acid residues at positions 350-357 of the wild type HPV33 L1 protein.

11. The mutated HPV58 L1 protein according to claim 1 , wherein the mutated HPV58 L1 protein has an amino acid sequence selected from the group consisting of: SEQ ID NOs: 4 and 10.

12. The method according to claim 7 , wherein the host cell is E. coli.

13. The method according to claim 9 , wherein: the HPV infection is HPV58 infection and/or HPV33 infection; and/or, the disease caused by HPV infection is selected from the group consisting of cervical cancer and condyloma acuminatum.

14. A mutated HPV58 L1 protein, wherein as compared with a wild type HPV58 L1 protein, the mutated HPV58 L1 protein has the following mutations:
(1) N-terminal truncation of 5-70 amino acids; and
(2)(a) substitution of amino acid residues at positions 80-87 of the wild type HPV58 L1 protein with amino acid residues at the corresponding positions of a wild-type HPV33 L1 protein; and
(3)(a) substitution of amino acid residues at positions 375-383 of the wild type HPV58 L1 protein with the amino acid residues at the corresponding positions of a wild type HPV52 L1 protein; or
(3)(b) substitution of amino acid residues at positions 144-168 of the wild type HPV58 L1 protein with amino acid residues at the corresponding positions of a wild type HPV52 L1 protein;
wherein said wild type HPV58 L1 protein has an amino acid sequence as set forth in SEQ ID NO: 1 or a sequence having 95% or greater identity to SEQ ID NO:1; wherein said wild type HPV33 L1 protein has an amino acid sequence as set forth in SEQ ID NO: 2 or a sequence having 95% or greater identity to SEQ ID NO:2; and wherein said wild type HPV52 L1 protein has an amino acid sequence as set forth in SEQ ID NO: 3 or a sequence having 95% or greater identity to SEQ ID NO:3.

15. The mutated HPV58 L1 protein according to claim 14 , wherein the mutated HPV58 L1 protein comprises one or more of the following items:
(i) the mutated HPV58 L1 protein has 5, 15, 27, 35, 40, 60 or 70 amino acids truncated at N-terminus, as compared with a wild type HPV58 L1 protein;
(ii) the amino acid residues at the corresponding positions as described in claim 14 , section (2) are amino acid residues at positions 54-61 of the wild type HPV33 L1 protein;
(iii) the amino acid residues at the corresponding positions as described in claim 14 , section (3)(a) are amino acid residues at positions 380-388 of a wild type HPV52 L1 protein; and
(iv) the amino acid residues at the corresponding positions as described in in claim 14 , section (3)(b) are amino acid residues at positions 146-170 of a wild type HPV52 L1 protein.

16. The mutated HPV58 L1 protein according to claim 14 , wherein the mutated HPV58 L1 protein has an amino acid sequence selected from the group consisting of: SEQ ID NOs: 11 and 14.

17. An isolated nucleic acid, encoding the mutated HPV58 L1 protein according to claim 14 .

18. A vector comprising the isolated nucleic acid according to claim 17 .

19. An isolated host cell comprising the isolated nucleic acid according to claim 17 and/or a vector comprising the isolated nucleic acid.

20. A HPV virus-like particle, comprising or consisting of the mutated HPV58 L1 protein according to claim 14 .

21. A pharmaceutical composition or vaccine, comprising the HPV virus-like particle according to claim 20 , and a pharmaceutically acceptable carrier and/or excipient.

22. A method for preparing the mutated HPV58 L1 protein according to claim 14 , comprising expressing the mutated HPV58 L1 protein in a host cell, and then recovering the mutated HPV58 L1 protein from a culture of the host cell.

23. The method according to claim 22 , wherein the host cell is E. coli.

24. A method for preparing a vaccine, comprising combining the HPV virus like particle according to claim 20 with a pharmaceutically acceptable carrier and/or excipient.

25. A method for preventing HPV infection or a disease caused by HPV infection, comprising administering to a subject a prophylactically effective amount of the HPV virus-like particle according to claim 20 or a pharmaceutical composition or vaccine comprising the HPV virus-like particle and a pharmaceutically acceptable carrier and/or excipient.

26. The method according to claim 25 , wherein the HPV infection is selected from one or more of the following: HPV58 infection, HPV33 infection and HPV52 infection; and/or, the disease caused by HPV infection is selected from the group consisting of cervical cancer and condyloma acuminatum.

Assignees

• Univ Xiamen
• Xiamen Innovax Biotech Co Ltd

Inventors

• Li Shaowei
• Li Zhihai
• Song Shuo
• He Maozhou
• Gu Ying
• Xia Ningshao

Classifications

• C12N2710/20023

  Virus like particles [VLP] · CPC title

• C07K14/005 Primary

  from viruses · CPC title

• A61P31/20

  for DNA viruses · CPC title

• C12N7/04

  Inactivation or attenuation; Producing viral sub-units · CPC title

• C12N15/70

  Vectors or expression systems specially adapted for E. coli · CPC title