Anti-HLA-G antibodies and use thereof
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a) a modified human HLA-G β2M MHC I complex, wherein the HLA-G specific amino acids have been replaced by HLA-A consensus amino acids, the complex comprising SEQ ID NO:40;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 .