Anti-HLA-G antibodies and use thereof

Abstract

The present invention relates antibodies that bind to human HLA-G, multispecific antibodies thereof, their preparation, formulations and methods of using the same.

1 . An antibody that binds to human HLA-G comprising
 A) (a) a VH domain comprising (i) CDR-H1 comprising an amino acid sequence of SEQ ID NO:1, (ii) CDR-H2 comprising an amino acid sequence of SEQ ID NO:2, and (iii) CDR-H3 comprising an amino acid sequence of SEQ ID NO:3; and (b) a VL domain comprising (i) CDR-L1 comprising an amino acid sequence of SEQ ID NO:23; (ii) CDR-L2 comprising an amino acid sequence of SEQ ID NO:5 and (iii) CDR-L3 comprising an amino acid sequence of SEQ ID NO:6, or   B) (a) a VH domain comprising (i) CDR-H1 comprising an amino acid sequence of SEQ ID NO:1, (ii) CDR-H2 comprising an amino acid sequence of SEQ ID NO:2, and (iii) CDR-H3 comprising an amino acid sequence of SEQ ID NO:3; and (b) a VL domain comprising (i) CDR-L1 comprising an amino acid sequence of SEQ ID NO:25; (ii) CDR-L2 comprising an amino acid sequence of SEQ ID NO:5 and (iii) CDR-L3 comprising an amino acid sequence of SEQ ID NO:6.   
     
     
         2 . The antibody according to  claim 1 , wherein the antibody
 A) comprises a VH domain comprising an amino acid sequence of SEQ ID NO:7 and a VL domain comprising an amino acid sequence of SEQ ID NO:24; or   B) comprises a VH domain comprising an amino acid sequence of SEQ ID NO:7 and a VL domain comprising an amino acid sequence of SEQ ID NO:26.   
     
     
         3 . The antibody of  claim 1 , wherein the antibody comprises a Fc domain of human origin. 
     
     
         4 .- 25 . (canceled) 
     
     
         26 . The antibody of  claim 3 , wherein the Fc domain of human origin is an IgG isotype. 
     
     
         27 . The antibody of  claim 26 , wherein the IgG is an IgG1 isotype. 
     
     
         28 . The antibody of  claim 27 , wherein the antibody comprises a constant region of human origin, comprising a human CH1, CH2, CH3 and/or CL domain. 
     
     
         29 . The antibody of  claim 1 , wherein the antibody does not cross-react with:
 a) a modified human HLA-G β2M MHC I complex, wherein the HLA-G specific amino acids have been replaced by HLA-A consensus amino acids, the complex comprising SEQ ID NO:40;   b) a mouse H2Kd β2M MHC I complex comprising SEQ ID NO:41; or   c) a rat RT1A β2M MHC I complex comprising SEQ ID NO:43.   
     
     
         30 . The antibody of  claim 1 , wherein the antibody:
 a) inhibits ILT2 binding to HLA-G expressed on JEG3 cells (ATCC No. HTB36); or   b) binds to HLA-G expressed on JEG3 cells (ATCC No. HTB36) and inhibits ILT2 binding to HLA-G expressed on JEG-3 cells (ATCC No. HTB36).   
     
     
         31 . The antibody of  claim 1 , wherein the antibody is a multi-specific antibody. 
     
     
         32 . One or more isolated nucleic acids encoding the antibody of  claim 1 . 
     
     
         33 . A host cell comprising the one or more nucleic acids of  claim 32 . 
     
     
         34 . The host cell of  claim 33 , wherein the host cell is an eukaryotic cell. 
     
     
         35 . A method of producing the antibody encoded by the one or more nucleic acids of the host cell of  claim 33 , comprising culturing the host cell so that the antibody is produced. 
     
     
         36 . The method of  claim 35 , further comprising recovering the antibody from the host cell. 
     
     
         37 . A pharmaceutical formulation comprising the antibody of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         38 . A method of treating an individual having cancer comprising administering to the individual an effective amount of the antibody of  claim 1 . 
     
     
         39 . A bispecific antibody, comprising
 a first antigen binding moiety that binds to human HLA-G and comprises:   A) (a) a VH domain comprising (i) CDR-H1 comprising an amino acid sequence of SEQ ID NO:1, (ii) CDR-H2 comprising an amino acid sequence of SEQ ID NO:2, and (iii) CDR-H3 comprising an amino acid sequence of SEQ ID NO:3; and (b) a VL domain comprising (i) CDR-L1 comprising an amino acid sequence of SEQ ID NO:23; (ii) CDR-L2 comprising an amino acid sequence of SEQ ID NO:5 and (iii) CDR-L3 comprising an amino acid sequence of SEQ ID NO:6, or   B) (a) a VH domain comprising (i) CDR-H1 comprising an amino acid sequence of SEQ ID NO:1, (ii) CDR-H2 comprising an amino acid sequence of SEQ ID NO:2, and (iii) CDR-H3 comprising an amino acid sequence of SEQ ID NO:3; and (b) a VL domain comprising (i) CDR-L1 comprising an amino acid sequence of SEQ ID NO:25; (ii) CDR-L2 comprising an amino acid sequence of SEQ ID NO:5 and (iii) CDR-L3 comprising an amino acid sequence of SEQ ID NO:6; and   a second antigen binding moiety that binds to human CD3 and comprises:   C) (a) a VH domain comprising (i) CDR-H1 comprising an amino acid sequence of SEQ ID NO:52, (ii) CDR-H2 comprising an amino acid sequence of SEQ ID NO:53, and (iii) CDR-H3 comprising an amino acid sequence of SEQ ID NO:54; and (b) a VL domain comprising (i) CDR-L1 comprising an amino acid sequence of SEQ ID NO:55; (ii) CDR-L2 comprising an amino acid sequence of SEQ ID NO:56 and (iii) CDR-L3 comprising an amino acid sequence of SEQ ID NO:57, or   D) (a) a VH domain comprising (i) CDR-H1 comprising an amino acid sequence of SEQ ID NO:60, (ii) CDR-H2 comprising an amino acid sequence of SEQ ID NO:61, and (iii) CDR-H3 comprising an amino acid sequence of SEQ ID NO:62; and (b) a VL domain comprising (i) CDR-L1 comprising an amino acid sequence of SEQ ID NO:63; (ii) CDR-L2 comprising an amino acid sequence of SEQ ID NO:64 and (iii) CDR-L3 comprising an amino acid sequence of SEQ ID NO:65, or   E) (a) a VH domain comprising (i) CDR-H1 comprising an amino acid sequence of SEQ ID NO:68, (ii) CDR-H2 comprising an amino acid sequence of SEQ ID NO:69, and (iii) CDR-H3 comprising an amino acid sequence of SEQ ID NO:70; and (b) a VL domain comprising (i) CDR-L1 comprising an amino acid sequence of SEQ ID NO:71; (ii) CDR-L2 comprising an amino acid sequence of SEQ ID NO:72 and (iii) CDR-L3 comprising an amino acid sequence of SEQ ID NO:73.   
     
     
         40 . The bispecific antibody according to  claim 39 ,
 wherein the first antigen binding moiety comprises:   A) a VH domain comprising an amino acid sequence of SEQ ID NO:7 and a VL domain comprising an amino acid sequence of SEQ ID NO:24; or   B) a VH domain comprising an amino acid sequence of SEQ ID NO:7 and a VL domain comprising an amino acid sequence of SEQ ID NO:26,   and wherein the second antigen binding moiety comprises:   C) a VH domain comprising an amino acid sequence of SEQ ID NO:58 and a VL domain comprising an amino acid sequence of SEQ ID NO:59; or   D) a VH domain comprising an amino acid sequence of SEQ ID NO:66 and a VL domain comprising an amino acid sequence of SEQ ID NO:67; or   E) a VH domain comprising an amino acid sequence of SEQ ID NO:74 and a VL domain comprising an amino acid sequence of SEQ ID NO:75.   
     
     
         41 . The bispecific antibody according to  claim 40 , wherein the first antigen binding moiety comprises a VH domain comprising an amino acid sequence of SEQ ID NO:7 and a VL domain comprising an amino acid sequence of SEQ ID NO:24; and wherein the second antigen binding moiety comprises a VH domain comprising an amino acid sequence of SEQ ID NO:58 and a VL domain comprising an amino acid sequence of SEQ ID NO:59. 
     
     
         42 . The bispecific antibody according to  claim 40 , wherein the first antigen binding moiety comprises a VH domain comprising an amino acid sequence of SEQ ID NO:7 and a VL domain comprising an amino acid sequence of SEQ ID NO:24; and wherein the second antigen binding moiety comprises a VH domain comprising an amino acid sequence of SEQ ID NO:66 and a VL domain comprising an amino acid sequence of SEQ ID NO:67. 
     
     
         43 . The bispecific antibody according to  claim 40 , wherein the first antigen binding moiety comprises a VH domain comprising an amino acid sequence of SEQ ID NO:7 and a VL domain comprising an amino acid sequence of SEQ ID NO:24; and wherein the second antigen binding moiety comprises a VH domain comprising an amino acid sequence of SEQ ID NO:74 and a VL domain comprising an amino acid sequence of SEQ ID NO:75. 
     
     
         44 . The bispecific antibody of  claim 39 , wherein the bispecific antibody shows:
 a) inhibition of ILT2 or ILT4 binding to HLA-G;   b) antibody-mediated IFN gamma secretion by T cells on
 i) SKOV3 cells transfected with recombinant HLA-G (SKOV3 HLA-G); or 
 ii) JEG3 cells expressing endogenous HLA-G; 
   c) T cell-mediated cytotoxicity or tumor cell killing on
 i) SKOV3 cells transfected with recombinant HLA-G (SKOV 3HLA-G); or 
 ii) JEG3 cells expressing endogenous HLA-G; 
   d) in vivo anti-tumor efficacy or tumor regression in humanized NSG mice bearing SKOV3 human ovarian carcinoma transfected with recombinant HLA-G (SKOV3 HLA-G); or   e) in vivo anti-tumor efficacy or tumor in humanized NSG mice bearing human breast cancer PDX tumors (BC004).   
     
     
         45 . One or more isolated nucleic acids encoding the bispecific antibody of  claim 39 . 
     
     
         46 . A host cell comprising the one or more nucleic acids of  claim 45 . 
     
     
         47 . The host cell of  claim 46 , wherein the host cell is an eukaryotic cell. 
     
     
         48 . A method of producing the bispecific antibody encoded by the one or more nucleic acids of the host cell of  claim 46 , comprising culturing the host cell so that the bispecific antibody is produced. 
     
     
         49 . The method of  claim 48 , further comprising recovering the bispecific antibody from the host cell. 
     
     
         50 . A pharmaceutical formulation comprising the bispecific antibody of  claim 39  and a pharmaceutically acceptable carrier. 
     
     
         51 . A method of treating an individual having cancer comprising administering to the individual an effective amount of the bispecific antibody of  claim 39 .