Monoclonal antibody against human lag-3, method for preparing the same, and use thereof

Patent metadata

Field	Value
Publication number	US-2020407442-A1
Application number	US-201916976404-A
Country	US
Kind code	A1
Filing date	Feb 27, 2019
Priority date	Feb 28, 2018
Publication date	Dec 31, 2020
Grant date	—

Abstract

Provided are novel fully human monoclonal antibodies that bind to human LAG-3. It also provides the methods of hybridoma generation using humanized rats, the nucleic acid molecules encoding the anti-LAG-3 antibodies, expression vectors and host cells used for the expression of anti-LAG-3 antibodies. The invention further provides the methods for validating the function of antibodies in vitro. The antibodies of invention provide a potent agent for the treatment of multiple cancers via modulating human immune function.

First claim

1 . An isolated antibody or the antigen-binding portion thereof, wherein the isolated antibody or the antigen-binding portion thereof comprises:
A) one or more heavy chain CDRs (CDRHs) selected from the group consisting of:
(i) a CDRH1 with at least 90% sequence identity to a CDRH1 as set forth in one of the sequences selected from the group consisting of SEQ ID NOs: 1 and 7;
(ii) a CDRH2 with at least 90% sequence identity to a CDRH2 as set forth in one of the sequences selected from the group consisting of SEQ ID NOs: 2 and 8; and
(iii) a CDRH3 with at least 90% sequence identity to a CDRH3 as set forth in one of the sequences selected from the group consisting of SEQ ID NOs: 3 and 9;
B) one or more light chain CDRs (CDRLs) selected from at least one of the group consisting of:
(i) a CDRL1 with at least 90% 0 sequence identity to a CDRL1 as set forth in one of the sequences selected from the group consisting of SEQ ID NOs: 4 and 10;
(ii) a CDRL2 with at least 90% sequence identity to a CDRL2 as set forth in one of the sequences selected from the group consisting of SEQ ID NOs: 5 and 11; and
(iii) a CDRL3 with at least 90% sequence identity to a CDRL3 as set forth in one of the sequences selected from the group consisting of SEQ ID NOs: 6 and 12; or
C) one or more CDRHs of A) and one or more CDRLs of B).

2 . The isolated antibody or the antigen-binding portion thereof of claim 1 , wherein the isolated antibody or the antigen-binding portion thereof comprises:
A) one or more heavy chain CDRs (CDRHs) selected from the group consisting of:
(i) a CDRH1 selected from the group consisting of SEQ ID NOs: 1 and 7 or a CDRH1 that differs in amino acid sequence from the CDRH1 by an amino acid addition, deletion or substitution of not more than 2 amino acids;
(ii) a CDRH2 selected from the group consisting of SEQ ID NOs: 2 and 8 or a CDRH2 that differs in amino acid sequence from the CDRH2 by an amino acid addition, deletion or substitution of not more than 2 amino acids; and
(iii) a CDRH3 selected from the group consisting of SEQ ID NOs: 3 and 9 or a CDRH3 that differs in amino acid sequence from the CDRH3 by an amino acid addition, deletion or substitution of not more than 2 amino acids;
B) one or more light chain CDRs (CDRLs) selected from the group consisting of:
(i) a CDRL1 selected from the group consisting of SEQ ID NOs: 4 and 10 or a CDRL1 that differs in amino acid sequence from the CDRL1 by an amino acid addition, deletion or substitution of not more than 2 amino acids;
(ii) a CDRL2 selected from the group consisting of SEQ ID NOs: 5 and 11 or a CDRL2 that differs in amino acid sequence from the CDRL2 by an amino acid addition, deletion or substitution of not more than 2 amino acids; and
(iii) a CDRL3 selected from the group consisting of SEQ ID NOs: 6 and 12 or a CDRL3 that differs in amino acid sequence from the CDRL3 by an amino acid addition, deletion or substitution of not more than 2 amino acids; or
C) one or more CDRHs of A) and one or more CDRLs of B).

3 . The isolated antibody or the antigen-binding portion thereof of claim 1 , wherein the isolated antibody or the antigen-binding portion thereof comprises:
(a) a CDRH1 comprising or consisting of SEQ ID NO: 1; (b) a CDRH2 comprising or consisting of SEQ ID NO: 2; (c) a CDRH3 comprising or consisting of SEQ ID NO: 3; (d) a CDRL1 comprising or consisting of SEQ ID NO: 4; (e) a CDRL2 comprising or consisting of SEQ ID NO: 5; and (f) a CDRL3 comprising or consisting of SEQ ID NO: 6.

4 . The isolated antibody or the antigen-binding portion thereof of claim 1 , wherein the isolated antibody or the antigen-binding portion thereof comprises:
(a) a CDRH1 comprising or consisting of SEQ ID NO: 7; (b) a CDRH2 comprising or consisting of SEQ ID NO: 8; (c) a CDRH3 comprising or consisting of SEQ ID NO: 9; (d) a CDRL1 comprising or consisting of SEQ ID NO: 10; (e) a CDRL2 comprising or consisting of SEQ ID NO: 11; and (f) a CDRL3 comprising or consisting of SEQ ID NO: 12.

5 . The isolated antibody or the antigen-binding portion thereof of claim 1 , wherein the isolated antibody or the antigen-binding portion thereof comprises:
(A) a heavy chain variable region:
(i) comprising the amino acid sequence of SEQ ID NO: 13;
(ii) comprising an amino acid sequence at least 85%, 90%, or 95% identical to SEQ ID NO: 13; or
(iii) comprising an amino acid sequence with addition, deletion and/or substitution of one or more amino acids compared with SEQ ID NO: 13; and/or
(B) a light chain variable region:
(i) comprising the amino acid sequence of SEQ ID NO: 14;
(ii) comprising an amino acid sequence at least 85%, at least 90%, or at least 95% identical to SEQ ID NO: 14; or
(iii) comprising an amino acid sequence with addition, deletion and/or substitution of one or more amino acids compared with SEQ ID NO: 14.

6 . The isolated antibody or the antigen-binding portion thereof of claim 1 , wherein the isolated antibody or the antigen-binding portion thereof comprises:
(A) a heavy chain variable region:
(i) comprising the amino acid sequence of SEQ ID NO: 15;
(ii) comprising an amino acid sequence at least 85%, at least 90%, or at least 95% identical to SEQ ID NO: 15; or
(iii) comprising an amino acid sequence with addition, deletion and/or substitution of one or more amino acids compared with SEQ ID NO: 15; and/or
(B) a light chain variable region:
(i) comprising the amino acid sequence of SEQ ID NO: 16;
(ii) comprising an amino acid sequence at least 85%, at least 90%, or at least 95% identical to SEQ ID NO: 16; or
(iii) comprising an amino acid sequence with addition, deletion and/or substitution of one or more amino acids compared with SEQ ID NO: 16.

7 . The isolated antibody or the antigen-binding portion thereof of claim 1 , having one or more of the following properties:
(a) binds to human LAG-3 with a K D of 2×10 −10 M or less; (b) inhibits binding of LAG-3 to major histocompatibility (MHC) class II molecules; (c) inhibits binding of LAG-3 to fibrinogen-like protein 1 (FGL1) ligand molecules; (d) inhibits binding of LAG-3 to LSECtin and/or Galectin-3; (e) binds to human LAG-3 without cross-family reactions; or (f) has no cross-reactivity to human CD4.

8 . The isolated antibody or the antigen-binding portion thereof of claim 1 , wherein the antibody is a monoclonal antibody, for example, a fully human monoclonal antibody, for example, a fully human monoclonal antibody produced by a transgenic mammal, preferably a transgenic rat, more preferably a transgenic rat with recombinant immunoglobulin loci.

9 . An isolated nucleic acid molecule, comprising a nucleic acid sequence encoding the heavy chain variable region and/or the light chain variable region of the isolated antibody as defined in claim 1 , for example, a nucleic acid sequence as shown in SEQ ID NOs: 17-20.

10 . An expression vector comprising the nucleic acid molecule of claim 9 .

11 . A host cell comprising the expression vector of claim 10 .

12 . A pharmaceutical composition comprising at least one antibody or antigen-binding portion thereof as defined in claim 1 and a pharmaceutically acceptable carrier.

13 . A method for preparing antibody or antigen-binding portion thereof as defined in claim 1 comprising the steps of:
expressing the antibody or antigen-binding portion thereof as defined in claim 1 in a host cell comprising an expression vector encoding the antibody or antigen-binding portion thereof; and
isolating the antibody or antigen-binding portion thereof from the host cell.

14 . A method of modulating an antigen-specific T ce

Assignees

• Wuxi Biologics Ireland Ltd

Inventors

• Zheng Yong
• Wu Qiong
• Li Jing

Classifications

• G01N33/5759

  involving compounds localised on the membrane of tumour or cancer cells · CPC title

• A61P35/00 Primary

  Antineoplastic agents · CPC title

• C07K2317/33

  Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity · CPC title

• C07K2317/21

  from primates, e.g. man · CPC title

• C07K2317/76

  Antagonist effect on antigen, e.g. neutralization or inhibition of binding · CPC title