Antibody against hepatitis B surface antigen and use thereof

• US 10,689,434 B2
• Filed: 10/09/2016
• Issued: 06/23/2020
• Est. Priority Date: 10/09/2015
• Status: Active Grant

• Current Assignee: Yangshengtang Company Limited (Shan Shan Zhong,), Xiamen University
• Original Assignee: Xiamen University
• Inventors: Luo, Wenxin, Zhou, Bing, Zhang, Juan, Yuan, Quan, Zhang, Tianying, Zhang, Jun, Xia, Ningshao
• Primary Examiner(s): Foley, Shanon A.
• Application Number:
  US15/766,593
• Publication Number:
  US 20190389939A1
• Time in Patent Office: 1,353 Days
• Field of Search: None
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: C07K 16/082 Hepadnaviridae, e.g. hepatitis B virus
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2317/94 Stability, e.g. half-life, pH, temperature or enzyme-resistance

Abstract

The invention provides an antibody (in particular, a humanized antibody) against hepatitis B surface antigen (HBsAg), a nucleic acid molecule encoding the same, a method for preparing the same, and a pharmaceutical composition comprising the same. The invention also provides use of the antibody and pharmaceutical composition. The antibody and pharmaceutical composition according to the invention can be used for preventing and/or treating HBV infection or a disease associated with HBV infection (such as Hepatitis B), for neutralizing HBV virulence in a subject (such as human), or for reducing the serum level of HBV DNA and/or HBsAg in a subject.

Claims

1. An antibody or an antigen binding fragment thereof, which can specifically bind to HBsAg, comprising:

(a) three complementarity determining regions (CDRs) of heavy chain variable region (VH) selected from the group consisting of;

(i) VH CDR1, consisting of the following sequence;

SEQ ID NO;

3, or a sequence that differs from SEQ ID NO;

3 by 1 or 2 substitutions selected from the group consisting of;

(01) R or Y at H31; and

(02) W at H32;

(ii) VH CDR2, consisting of the following sequence;

SEQ ID NO;

4, or a sequence that differs from SEQ ID NO;

4 by 1, 2 or 3 substitutions selected from the group consisting of;

(12) T at H56;

(13) V or N at H57; and

(14) L at H58;

(iii) VH CDR3, consisting of the following sequence;

SEQ ID NO;

5

and

(b) three CDRs of light chain variable region (VL) selected from the group consisting of;

(iv) VL CDR1, consisting of the following sequence;

SEQ ID NO;

6, or a sequence that differs from SEQ ID NO;

6 by 1 or 2 substitutions selected from the group consisting of;

(29) P or T at L27; and

(32) S or N at L30;

(v) VL CDR2, consisting of the following sequence;

SEQ ID NO;

7, and

(vi) VL CDR3, consisting of the following sequence;

SEQ ID NO;

8;

wherein, the amino acid positions mentioned above are numbered according to Kabat numbering system;

and, the antibody or an antigen binding fragment thereof is humanized, and has a humanization degree of at least 85%.

2. The antibody or an antigen binding fragment thereof according to claim 1, wherein the antibody or an antigen binding fragment thereof is characterized by any one or more of (i), (ii) and (iii):

(i) the VH CDR1 has a sequence selected from the group consisting of;

3. The antibody or an antigen binding fragment thereof according to claim 1, wherein

4. The antibody or an antigen binding fragment thereof according to claim 1, wherein the antibody comprises a VL comprising:

(a) VL CDR1 as set forth in RSSQSLVHSYGDTYLH (SEQ ID NO;

     6);

VL CDR2 as set forth in KVSNRFS (SEQ ID NO;

     7); and

VL CDR3 as set forth in SQNTHVPYT (SEQ ID NO;

     8);

(k) VL CDR1 as set forth in RSSQSLVHTYGNTYLH (SEQ ID NO;

     235);

VL CDR2 as set forth in KVSNRFS (SEQ ID NO;

     7); and

VL CDR3 as set forth in SQNTHVPYT (SEQ ID NO;

     8);

or

(l) VL CDR1 as set forth in RSSQSLVHPYGSTYLH (SEQ ID NO;

     236);

VL CDR2 as set forth in KVSNRFS (SEQ ID NO;

     7); and

VL CDR3 as set forth in SQNTHVPYT (SEQ ID NO;

     8).

5. The antibody or an antigen binding fragment thereof according to claim 4, wherein the antibody comprises a VH comprising:

(a) VH CDR1 as set forth in SGYHWN (SEQ ID NO;

     3);

VH CDR2 as set forth in YISYDGSDHYNPSLEN (SEQ ID NO;

     4); and

VH CDR3 as set forth in GFDH (SEQ ID NO;

     5);

(j) VH CDR1 as set forth in RWYHWN (SEQ ID NO;

     126);

VH CDR2 as set forth in YISYDGTVLYNPSLEN (SEQ ID NO;

     146); and

VH CDR3 as set forth in GFDH (SEQ ID NO;

     5);

(z) VH CDR1 as set forth in YGYHWN (SEQ ID NO;

     124);

VH CDR2 as set forth in YISYDGSVLYNPSLEN (SEQ ID NO;

     149); and

VH CDR3 as set forth in GFDH (SEQ ID NO;

     5);

(ad) VH CDR1 as set forth in RGYHWN (SEQ ID NO;

     121);

VH CDR2 as set forth in YISYDGTNLYNPSLEN (SEQ ID NO;

     151); and

VH CDR3 as set forth in GFDH (SEQ ID NO;

     5);

QSYNPSLKG (SEQ ID NO;

     157); and

VH CDR3 as set forth in GFDH (SEQ ID NO;

     5);

(ap) VH CDR1 as set forth in RGYHWN (SEQ ID NO;

     121);

VH CDR2 as set forth in YISYDGSVLYNPSLKS (SEQ ID NO;

     155); and

VH CDR3 as set forth in GFDH (SEQ ID NO;

     5);

or,

(aq) VH CDR1 as set forth in SGYHWN (SEQ ID NO;

     3);

VH CDR2 as set forth in YISYDGSVLYNPSLKS (SEQ ID NO;

     155); and

VH CDR3 as set forth in GFDH (SEQ ID NO;

     5).

6. The antibody or an antigen binding fragment thereof according to claim 1, wherein the antibody comprises 6 CDRs from heavy chain and light chain variable regions of an antibody selected from the group consisting of:

7. The antibody or an antigen binding fragment thereof according to claim 1, wherein the antibody has:

(i) a heavy chain variable region, which has an amino acid sequence identity of at least 80% with a heavy chain variable region selected from the group consisting of;

heavy chain variable regions set forth in SEQ ID NOs;

20, 36, 41, 69, 72, 73, 91;

and

(ii) a light chain variable region, which has an amino acid sequence identity of at least 80%, with a light chain variable region selected from the group consisting of;

light chain variable regions set forth in SEQ ID NOs;

187, 189, 201, 202, 205.

8. The antibody or an antigen binding fragment thereof according to claim 7, wherein, the antibody comprises:

(8) VH as set forth in SEQ ID NO;

69 and VL as set forth in SEQ ID NO;

189;

(10) VH as set forth in SEQ ID NO;

73 and VL as set forth in SEQ ID NO;

202;

(16) VH as set forth in SEQ ID NO;

91 and VL as set forth in SEQ ID NO;

205;

(18) VH as set forth in SEQ ID NO;

36 and VL as set forth in SEQ ID NO;

187;

(19) VH as set forth in SEQ ID NO;

36 and VL as set forth in SEQ ID NO;

189;

(27) VH as set forth in SEQ ID NO;

20 and VL as set forth in SEQ ID NO;

187;

or,

(49) VH as set forth in SEQ ID NO;

41 and VL as set forth in SEQ ID NO;

187.

9. The antibody or an antigen binding fragment thereof according to claim 1, wherein the antibody or an antigen binding fragment thereof is characterized by any one or more of (i), (ii) and (iii):

(i) the antibody or an antigen binding fragment thereof is selected from the group consisting of scFv, Fab, Fab′

, (Fab′

)₂, Fv fragment, diabody, bispecific antibody, and polyspecific antibody;

(ii) the antibody or an antigen binding fragment thereof is an antibody of IgG isotype or an antigen binding fragment thereof;

(iii) the antibody or an antigen binding fragment thereof is labeled.

10. The antibody or an antigen binding fragment thereof according to claim 1, wherein the antibody or an antigen binding fragment thereof is characterized by any one or more of (i) and (ii):

(i) the antibody or an antigen binding fragment thereof has a humanization degree of at least 85%;

(ii) the antibody or an antigen binding fragment thereof comprises no more than 20 murine amino acid residues in framework region.

11. An isolated nucleic acid molecule, encoding the antibody or an antigen binding fragment thereof according to claim 1.

12. A vector, comprising the nucleic acid molecule according to claim 11.

13. A host cell comprising the nucleic acid molecule according to claim 11 or a vector comprising the nucleic acid molecule.

14. A method for preparing the antibody or an antigen binding fragment thereof according to claim 1, comprising culturing a host cell under a condition allowing expression of the antibody or an antigen binding fragment thereof, and recovering the antibody or an antigen binding fragment thereof from a culture of the cultured host cell;

wherein the host cell comprises a nucleic acid molecule encoding the antibody or an antigen binding fragment thereof or a vector comprising the nucleic acid molecule.

15. A pharmaceutical composition, comprising the antibody or an antigen binding fragment thereof according to claim 1, and a pharmaceutically acceptable carrier and/or excipient.

16. A method for preventing or treating HBV infection or a disease associated with HBV infection in a subject, for neutralizing HBV virulence in a subject, and/or for reducing the serum level of HBV DNA and/or HBsAg in a subject, comprising administering to a subject in need thereof an effective amount of the antibody or an antigen binding fragment thereof according to claim 1 or a pharmaceutical composition comprising the antibody or an antigen binding fragment thereof.

17. The method according to claim 16, wherein the method is characterized by any one or more of (i) and (ii):

(i) the disease associated with HBV infection is Hepatitis B;

(ii) the subject is human.