Mutant of L1 protein of human papillomavirus type 16

Patent metadata

Field	Value
Publication number	US-11213580-B2
Application number	US-201816630673-A
Country	US
Kind code	B2
Filing date	Jul 13, 2018
Priority date	Jul 14, 2017
Publication date	Jan 4, 2022
Grant date	Jan 4, 2022

Abstract

Provided are a mutated HPV16 L1 protein (or a variant thereof), a sequence encoding the same and a method for preparing the same, as well as a virus-like particle comprising the same. The protein (or variant thereof) and the virus-like particle are capable of inducing neutralizing antibodies against at least two types of HPV (e.g., HPV16 and HPV35, or HPV16, HPV35, and HPV31), 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. Also provided are a use of the above protein and virus-like particle in the manufacture of a pharmaceutical composition or 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

What is claimed is:

1. A mutated HPV16 L1 protein or a variant thereof, wherein the mutated HPV16 L1 protein has the following mutations compared to a wild-type HPV16 L1 protein:
(1) a N-terminal truncation of 4-50 amino acids; and
(2) a substitution of amino acid residues at positions 292-316 of the wild-type HPV16 L1 protein with amino acid residues at the corresponding positions of a L1 protein of a second type of wild-type HPV;
optionally, the mutated HPV16 L1 protein further has the following mutation:
(3) a substitution of amino acid residues at positions 76-87 of the wild-type HPV16 L1 protein with amino acid residues at the corresponding positions of a L1 protein of a third type of wild-type HPV;
(4) a substitution of amino acid residues at positions 152-167 of the wild-type HPV16 L1 protein with amino acid residues at the corresponding positions of a L1 protein of a third type of wild-type HPV; or,
(5) a substitution of amino acid residues at positions 202-207 of the wild-type HPV16 L1 protein with amino acid residues at the corresponding positions of a L1 protein of a third type of wild-type HPV;
and, the variant differs from the mutated HPV16 L1 protein only by substitution, addition or deletion of 1, 2, 3, 4, 5, 6, 7, 8, or 9 amino acids, and maintains the function of the mutated HPV16 L1 protein, including an ability of inducing neutralizing antibodies against at least two types of HPV.

2. An isolated nucleic acid, which encodes the mutated HPV16 L1 protein or variant thereof according to claim 1 .

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

4. A 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, which comprises or consists of the mutated HPV16 L1 protein or variant thereof according to claim 1 .

6. A composition, which comprises:
(i) the mutated HPV16 L1 protein or a variant thereof according to claim 1 , or
(ii) an isolated nucleic acid encoding the mutated HPV16 L1 protein or variant thereof as described in (i), or
(iii) a vector comprising the isolated nucleic acid as described in (ii), or
(iv) a host cell comprising the isolated nucleic acid as described in (ii) or the vector as described in (iii), or
(v) a HPV virus-like particle comprising or consisting of the mutated HPV16 L1 protein or variant thereof as described in (i).

7. A pharmaceutical composition or vaccine, which comprises the HPV virus-like particle according to claim 5 , and optionally further comprises a pharmaceutically acceptable carrier and/or excipient.

8. A method for preparing the mutated HPV16 L1 protein or variant thereof according to claim 1 , which comprises expressing the mutated HPV16 L1 protein or variant thereof in a host cell, and then recovering the mutated HPV16 L1 protein or a variant thereof from a culture of the host cell.

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

10. A method for preventing HPV infection or a disease caused by HPV infection, which comprises 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 optionally a pharmaceutically acceptable carrier and/or excipient.

11. The mutated HPV16 L1 protein or variant thereof according to claim 1 , wherein the mutated HPV16 L1 protein is characterized by one or more of the following items:
(i) the mutated HPV16 L1 protein has a N-terminal truncation of 4, 6, 8, 10, 20, 30 or 40 amino acids compared with a wild-type HPV16 L1 protein;
(ii) the second type of wild-type HPV is HPV35;
(iii) the third type of wild-type HPV is HPV31;
(iv) the amino acid residues at the corresponding positions described in (2) are the amino acid residues at positions 266-288 of the wild-type HPV35 L1 protein;
(v) the amino acid residues at the corresponding positions described in (3) are the amino acid residues at positions 50-62 of the wild-type HPV31 L1 protein;
(vi) the amino acid residues at the corresponding positions described in (4) are the amino acid residues at positions 127-142 of the wild-type HPV31 L1 protein;
(vii) the amino acid residues at the corresponding positions described in (5) are the amino acid residues at positions 177-182 of the wild-type HPV31 L1 protein;
(viii) the wild-type HPV16 L1 protein has an amino acid sequence set forth in SEQ ID NO: 1;
(ix) the wild-type HPV35 L1 protein has an amino acid sequence set forth in SEQ ID NO: 2; and
(x) the wild-type HPV31 L1 protein has an amino acid sequence set forth in SEQ ID NO: 3.

12. The mutated HPV16 L1 protein or variant thereof according to claim 1 , wherein the mutated HPV16 L1 protein has an amino acid sequence selected from the group consisting of SEQ ID NOs: 7, 9, 10, and 11.

13. The pharmaceutical composition or vaccine according to claim 7 , wherein the HPV virus-like particle is present in an amount effective for preventing HPV infection or a disease caused by HPV infection.

14. The pharmaceutical composition or vaccine according to claim 13 , wherein: the HPV infection is infection by one or more HPV types; and/or, the disease caused by HPV infection is selected from the group consisting of a cervical cancer and a condyloma acuminatum.

15. The pharmaceutical composition or vaccine according to claim 13 , wherein the HPV infection is one or more selected from the group consisting of the following: HPV 16 infection, HPV 35 infection, and HPV 31 infection.

16. The method according to claim 8 , wherein the host cell is E. coli.

17. The method according to claim 8 , wherein the method comprises: expressing the mutated HPV16 L1 protein or a variant thereof in E. coli , and then obtaining the mutated HPV16 L1 protein or a variant thereof by purifying a lysate supernatant of the E. coli.

18. The method according to claim 10 , wherein: the HPV infection is infection by one or more HPV types; and/or, the disease caused by HPV infection is selected from the group consisting of cervical cancer and condyloma acuminatum.

19. The method according to claim 10 , wherein the HPV infection is one or more selected from the group consisting of the following: HPV 16 infection, HPV 35 infection, and HPV 31 infection.

Assignees

• Univ Xiamen
• Xiamen Innovax Biotech Co Ltd

Inventors

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

Classifications

• C12N2710/20034

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title

• C12N2710/20023

  Virus like particles [VLP] · CPC title

• C12N2710/20022

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• C07K14/005 Primary

  from viruses · CPC title

• A61K2039/5258

  Virus-like particles · CPC title