Polypeptides and antibodies for treating HBV infection and related diseases

Patent metadata

Field	Value
Publication number	US-10246494-B2
Application number	US-201715649967-A
Country	US
Kind code	B2
Filing date	Jul 14, 2017
Priority date	Jun 11, 2012
Publication date	Apr 2, 2019
Grant date	Apr 2, 2019

Abstract

The present invention relates to epitope peptides (or mutants thereof) for treating hepatitis B virus infection, recombinant proteins comprising such epitope peptides (or mutants thereof) and carrier proteins, and uses of such epitope peptides (or mutants thereof) and recombinant proteins. The present invention also relates to antibodies against such epitope peptides, cell lines producing said antibodies, and uses thereof. Furthermore, the present invention relates to vaccines or pharmaceutical compositions for treating or alleviating one or more symptoms associated with hepatitis B virus infection, which comprise the recombinant proteins or antibodies according to the invention, respectively.

First claim

The invention claimed is:

1. A monoclonal antibody or an antigen binding fragment thereof, wherein the monoclonal antibody specifically binds to positions 119 to 125 of HBsAg protein of HBV, and is selected from one of the following monoclonal antibodies:
1) the monoclonal antibody produced by the hybridoma cell line HBs-E6F6, wherein the hybridoma cell line HBs-E6F6 is deposited in China Center for Type Culture Collection (CCTCC), with a deposition number of CCTCC NO. C201270;
2) the monoclonal antibody produced by the hybridoma cell line HBs-E7G11, wherein the hybridoma cell line HBs-E7G11 is deposited in China Center for Type Culture Collection (CCTCC), with a deposition number of CCTCC NO. C201271;
3) the monoclonal antibody produced by the hybridoma cell line HBs-G12F5, wherein the hybridoma cell line HBs-G12F5 is deposited in China Center for Type Culture Collection (CCTCC), with a deposition number of CCTCC NO. C201272;
4) the monoclonal antibody produced by the monoclonal antibody produced by hybridoma cell line HBs-E13C5, wherein the hybridoma cell line HBs-E13C5 is deposited in China Center for Type Culture Collection (CCTCC), with a deposition number of CCTCC NO. C201273 and
5) the monoclonal antibody or an antigen binding fragment selected from a humanized antibody, a chimeric antibody or a single chain antibody made from any of the monoclonal antibodies as described in 1-4 above.

2. The monoclonal antibody or antigen binding fragment thereof of claim 1 , wherein the monoclonal antibody or antigen binding fragment thereof has one or more of the following features:
(1) the antigen binding fragment thereof is selected from a group consisting of Fab, Fab′, F(ab′) 2 , Fd, Fv, dAb, complementary determining region fragment, or a single chain antibody;
(2) the monoclonal antibody binds to HBsAg protein with a K D of less than about 10 −5 M, or less than about 10 −6 M, 10 −7 M, 10 −8 M, 10 −9 M, 10 −10 M or less;
(3) the monoclonal antibody comprises non-CDR region, and the non-CDR region is from species other than murine species; and
(4) the monoclonal antibody is capable of reducing serum level of HBV DNA and/or HBsAg in a subject.

3. The monoclonal antibody or antigen binding fragment thereof of claim 2 , wherein the single chain antibody is scFv, and/or, the chimeric antibody is a human-mouse chimeric antibody.

4. The monoclonal antibody or antigen binding fragment thereof of claim 2 , wherein the non-CDR region is from human antibody.

5. An isolated nucleic acid molecule, encoding the monoclonal antibody or antigen binding fragment thereof of claim 1 .

6. A vector, comprising the isolated nucleic acid molecule of claim 5 .

7. A host cell, comprising the isolated nucleic acid molecule according to claim 5 or a vector comprising the isolated nucleic acid molecule.

8. A method for producing the monoclonal antibody or antigen binding fragment thereof of claim 1 , comprising
1) expressing an isolated nucleic acid molecule encoding the monoclonal antibody or antigen binding fragment thereof in a host cell, to produce the monoclonal antibody or antigen binding fragment thereof; and
2) isolating the monoclonal antibody or antigen binding fragment thereof.

9. A hybridoma cell line, selected from:
1) hybridoma cell line HBs-E6F6, deposited in China Center for Type Culture Collection (CCTCC), with a deposition number of CCTCC NO. C201270;
2) hybridoma cell line HBs-E7G11, deposited in China Center for Type Culture Collection (CCTCC), with a deposition number of CCTCC NO. C201271;
3) hybridoma cell line HBs-G12F5, deposited in China Center for Type Culture Collection (CCTCC), with a deposition number of CCTCC NO. C201272; and
4) hybridoma cell line HBs-E13C5, deposited in China Center for Type Culture Collection (CCTCC), with a deposition number of CCTCC NO. C201273.

10. A kit for detecting the presence or level of HBsAg protein in a sample or for diagnosing whether a subject is infected by HBV, comprising the monoclonal antibody or antigen binding fragment thereof of claim 1 .

11. The kit according to claim 10 , wherein the monoclonal antibody or antigen binding fragment thereof further comprises a detectable marker.

12. The kit according to claim 11 , wherein the detectable marker comprised in the monoclonal antibody or antigen binding fragment thereof is selected from a radioisotope, a fluorescent substance, a luminescent substance, a chromophoric substance and an enzyme.

13. The kit according to claim 10 , wherein the kit further comprises a second antibody that specifically recognizes the monoclonal antibody or antigen binding fragment thereof; optionally, the second antibody further comprises a detectable marker.

14. The kit according to claim 13 , wherein the detectable marker comprised in the second antibody is selected from a radioisotope, a fluorescent substance, a luminescent substance, a chromophoric substance and an enzyme.

15. A method for detecting the presence or level of HBsAg protein in a sample, comprising contacting the monoclonal antibody or antigen binding fragment thereof of claim 1 with the sample, and detecting any binding between the monoclonal antibody or antigen binding fragment thereof and HBsAg protein.

16. A method for diagnosing whether a subject is infected by HBV, comprising (1) contacting the monoclonal antibody or antigen binding fragment thereof of claim 1 with a sample from a subject that is suspected to be infected by HBV, and (2) detecting the presence of HBV in the sample by detecting any binding between the monoclonal antibody or antigen binding fragment thereof and HBsAg protein.

17. A pharmaceutical composition comprising the monoclonal antibody or antigen binding fragment thereof of claim 1 , and a pharmaceutically acceptable carrier and/or excipient.

18. A method for preventing or treating HBV infection or a disease associated with HBV infection in a subject, comprising administering a prophylactically or therapeutically effective amount of the monoclonal antibody or antigen binding fragment thereof of claim 1 to a subject in need thereof.

19. The method according to claim 18 , wherein the disease associated with HBV infection is hepatitis B.

20. A method for reducing serum level of HBV DNA and/or HBsAg in a subject, comprising administering to a subject infected with HBV and in need of reducing serum level of HBV DNA and/or HBsAg an effective amount of one of the following:
(1) the monoclonal antibody or antigen binding fragment thereof of claim 1 ; and
(2) a monoclonal antibody or an antigen binding fragment thereof, capable of blocking the binding of HBsAg protein to the monoclonal antibody or antigen binding fragment thereof as defined in (1) by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 99%.

21. The monoclonal antibody or antigen binding fragment thereof according to claim 1 , wherein the antibody is a humanized antibody.

22. The monoclonal antibody or antigen binding fragment thereof according to claim 1 , wherein the antibody is a chimeric antibody.

23. The monoclonal antibody or antigen binding fragment thereof according to claim 1 , wherein the antibody is a single chain antibody.

Assignees

• Univ Xiamen
• Xiamen Innovax Biotech Co Ltd
• Yang Sheng Tang Co Ltd

Inventors

• Yuan Quan
• Zhang Tianying
• Luo Wenxin
• Chen Yixin
• Zhang Jun
• Xia Ningshao

Classifications

• A61P37/04

  Immunostimulants · CPC title

• A61P31/20

  for DNA viruses · CPC title

• A61P1/16

  for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics · CPC title

• A61K2039/505

  comprising antibodies · CPC title

• C07K14/005 Primary

  from viruses · CPC title