MUC1 and CD16A Antibodies and Methods of Use
The present disclosure provides for multi-specific antibodies and antigen-binding fragments thereof that bind to human MUC1 and CD16A, pharmaceutical compositions comprising said antibodies, and use of the antibodies or the compositions for treating a disease, such as cancer.
1 . A multi-specific antibody or antigen-binding fragment thereof, comprising a first antigen binding domain that specifically binds to human MUC1 and a second antigen binding domain that specifically binds to human CD16A.
2 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the second antigen binding domain has high selectivity over human CD16B.
3 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the first antigen binding domain that specifically binds to human MUC1 comprises:
(i). a heavy chain variable region (VH) that comprises (a) a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 24, (b) a HCDR2 of SEQ ID NO: 25, and (c) a HCDR3 of SEQ ID NO: 26, and a light chain variable region (VL) that comprises: (d) a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 27, (e) a LCDR2 of SEQ ID NO: 28, and (f) a LCDR3 of SEQ ID NO: 29;
(ii). a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 24, (b) a HCDR2 of SEQ ID NO: 25, and (c) a HCDR3 of SEQ ID NO: 26, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 27, (e) a LCDR2 of SEQ ID NO: 65, and (f) a LCDR3 of SEQ ID NO: 29;
(iii). a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 4, (b) a HCDR2 of SEQ ID NO: 5, and (c) a HCDR3 of SEQ ID NO: 6, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 7, (e) a LCDR2 of SEQ ID NO: 8, and (f) a LCDR3 of SEQ ID NO: 9; or
(iv). a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 14, (b) a HCDR2 of SEQ ID NO: 15, and (c) a HCDR3 of SEQ ID NO: 16, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 17, (e) a LCDR2 of SEQ ID NO: 18, and (f) a LCDR3 of SEQ ID NO: 19.
4 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the first antigen binding domain comprises:
(i). a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 30, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 31;
(ii). a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 61, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 62;
(iii). a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO: 61, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 66;
(iv). a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 61, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 68;
(v). a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO: 10, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 11; or
(vi). a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO: 20, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 21.
5 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein one, two, three, four, five, six, seven, eight, nine, or ten amino acids within one or more of SEQ ID NO: 30, 31, 61, 62, 66, 68, 10, 11, 20, and 21 have been inserted, deleted, or substituted.
6 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the first antigen binding domain comprises:
(i). a heavy chain variable region (VH) that comprises SEQ ID NO: 30, and a light chain variable region (VL) that comprises SEQ ID NO: 31;
(ii). a heavy chain variable region (VH) that comprises SEQ ID NO: 61, and a light chain variable region (VL) that comprises SEQ ID NO: 62;
(iii). a heavy chain variable region (VH) that comprises SEQ ID NO: 61, and a light chain variable region (VL) that comprises SEQ ID NO: 66;
(iv). a heavy chain variable region (VH) that comprises SEQ ID NO: 61, and a light chain variable region (VL) that comprises SEQ ID NO: 68;
(v). a heavy chain variable region (VH) that comprises SEQ ID NO: 10, and a light chain variable region (VL) that comprises SEQ ID NO: 11; or
(vi). a heavy chain variable region (VH) that comprises SEQ ID NO: 20, and a light chain variable region (VL) that comprises SEQ ID NO: 21.
7 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the second antigen binding domain that specifically binds to human CD16A comprises:
(i). a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 109, (b) a HCDR2 of SEQ ID NO: 110, and (c) a HCDR3 of SEQ ID NO: 111; or
(ii). a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 109, (b) a HCDR2 of SEQ ID NO: 114, and (c) a HCDR3 of SEQ ID NO: 111.
8 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the second antigen binding domain comprises:
(i). a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 112;
(ii). a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 115;
(iii). a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 117; or
(iv). a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 119.
9 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein one, two, three, four, five, six, seven, eight, nine, or ten amino acids within one or more of SEQ ID NO: 112, 115, 117, and 119 have been inserted, deleted, or substituted.
10 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the second antigen binding domain comprises:
(i). a heavy chain variable region (VH) that comprises SEQ ID NO: 112;
(ii). a heavy chain variable region (VH) that comprises SEQ ID NO: 115;
(iii). a heavy chain variable region (VH) that comprises SEQ ID NO: 117; or
(iv). a heavy chain variable region (VH) that comprises SEQ ID NO: 119.
11 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein;
(i). the first antigen binding domain that specifically binds to human MUC1 comprises: a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 24, (b) a HCDR2 of SEQ ID NO: 25, and (c) a HCDR3 of SEQ ID NO: 26, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 27, (e) a LCDR2 of SEQ ID NO: 28, and (f) a LCDR3 of SEQ ID NO: 29; and the second antigen binding domain that specifically binds to human CD16A comprises: (a) a HCDR1 of SEQ ID NO: 109, (b) a HCDR2 of SEQ ID NO: 110, and (c) a HCDR3 of SEQ ID NO: 111;
(ii). the first antigen binding domain that specifically binds to human MUC1 comprises: a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 24, (b) a HCDR2 of SEQ ID NO: 25, and (c) a HCDR3 of SEQ ID NO: 26, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 27, (e) a LCDR2 of SEQ ID NO: 65, and (f) a LCDR3 of SEQ ID NO: 29; and the second antigen binding domain that specifically binds to human CD16A comprises: (a) a HCDR1 of SEQ ID NO: 109, (b) a HCDR2 of SEQ ID NO: 110, and (c) a HCDR3 of SEQ ID NO: 111;
(iii). the first antigen binding domain that specifically binds to human MUC1 comprises: a heavy chain variable region (VH) hat comprises (a) a HCDR1 of SEQ ID NO: 4, (b) a HCDR2 of SEQ ID NO: 5, and (c) a HCDR3 of SEQ ID NO: 6, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 7, (e) a LCDR2 of SEQ ID NO: 8, and (f) a LCDR3 of SEQ ID NO: 9; and the second antigen binding domain that specifically binds to human CD16A comprises: (a) a HCDR1 of SEQ ID NO: 109, (b) a HCDR2 of SEQ ID NO: 110, and (c) a HCDR3 of SEQ ID NO: 111;
(iv). a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 14, (b) a HCDR2 of SEQ ID NO: 15, and (c) a HCDR3 of SEQ ID NO: 16, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 17, (e) a LCDR2 of SEQ ID NO: 18, and (f) a LCDR3 of SEQ ID NO: 19; and the second antigen binding domain that specifically binds to human CD16A comprises: (a) a HCDR1 of SEQ ID NO: 109, (b) a HCDR2 of SEQ ID NO: 110, and (c) a HCDR3 of SEQ ID NO: 111;
(v). the first antigen binding domain that specifically binds to human MUC1 comprises: a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 24, (b) a HCDR2 of SEQ ID NO: 25, and (c) a HCDR3 of SEQ ID NO: 26, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 27, (e) a LCDR2 of SEQ ID NO: 28, and (f) a LCDR3 of SEQ ID NO: 29; and the second antigen binding domain that specifically binds to human CD16A comprises: (a) a HCDR1 of SEQ ID NO: 109, (b) a HCDR2 of SEQ ID NO: 114, and (c) a HCDR3 of SEQ ID NO: 111;
(vi). the first antigen binding domain that specifically binds to human MUC1 comprises: a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 24, (b) a HCDR2 of SEQ ID NO: 25, and (c) a HCDR3 of SEQ ID NO: 26, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 27, (e) a LCDR2 of SEQ ID NO: 65, and (f) a LCDR3 of SEQ ID NO: 29; and the second antigen binding domain that specifically binds to human CD16A comprises: (a) a HCDR1 of SEQ ID NO: 109, (b) a HCDR2 of SEQ ID NO: 114, and (c) a HCDR3 of SEQ ID NO: 111;
(vii). the first antigen binding domain that specifically binds to human MUC1 comprises: a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 4, (b) a HCDR2 of SEQ ID NO: 5, and (c) a HCDR3 of SEQ ID NO: 6, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 7, (e) a LCDR2 of SEQ ID NO: 8, and (f) a LCDR3 of SEQ ID NO: 9; and the second antigen binding domain that specifically binds to human CD16A comprises: (a) a HCDR1 of SEQ ID NO: 109, (b) a HCDR2 of SEQ ID NO: 114, and (c) a HCDR3 of SEQ ID NO: 111; or
(viii). a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 14, (b) a HCDR2 of SEQ ID NO: 15, and (c) a HCDR3 of SEQ ID NO: 16, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 17, (e) a LCDR2 of SEQ ID NO: 18, and (f) a LCDR3 of SEQ ID NO: 19; and the second antigen binding domain that specifically binds to human CD16A comprises: (a) a HCDR1 of SEQ ID NO: 109, (b) a HCDR2 of SEQ ID NO: 114, and (c) a HCDR3 of SEQ ID NO: 111.
12 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein;
(i). the first antigen binding domain that specifically binds to human MUC1 comprises:
a) a heavy chain variable region (VH) that comprises SEQ ID NO: 30, and a light chain variable region (VL) that comprises SEQ ID NO: 31;
b) a heavy chain variable region (VH) that comprises SEQ ID NO: 61, and a light chain variable region (VL) that comprises SEQ ID NO: 62;
c) a heavy chain variable region (VH) that comprises SEQ ID NO: 61, and a light chain variable region (VL) that comprises SEQ ID NO: 66;
d) a heavy chain variable region (VH) that comprises SEQ ID NO: 61, and a light chain variable region (VL) that comprises SEQ ID NO: 68;
e) a heavy chain variable region (VH) that comprises SEQ ID NO: 10, and a light chain variable region (VL) that comprises SEQ ID NO: 11; or
f) a heavy chain variable region (VH) that comprises SEQ ID NO: 20, and a light chain variable region (VL) that comprises SEQ ID NO: 21;
(ii). and the second antigen binding domain that specifically binds to human CD16A comprises:
a) a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 109, (b) a HCDR2 of SEQ ID NO: 110, (c) a HCDR3 of SEQ ID NO: 111; or
b) a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 109, (b) a HCDR2 of SEQ ID NO: 114, (c) a HCDR3 of SEQ ID NO: 111.
13 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein;
(i). the first antigen binding domain that specifically binds to human MUC1 comprises:
a) a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 24, (b) a HCDR2 of SEQ ID NO: 25, and (c) a HCDR3 of SEQ ID NO: 26, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 27, (e) a LCDR2 of SEQ ID NO: 28, and (f) a LCDR3 of SEQ ID NO: 29;
b) a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 24, (b) a HCDR2 of SEQ ID NO: 25, and (c) a HCDR3 of SEQ ID NO: 26, and a light chain variable region (VL) hat comprises: (d) a LCDR1 of SEQ ID NO: 27, (e) a LCDR2 of SEQ ID NO: 65, and (f) a LCDR3 of SEQ ID NO: 29;
c) a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 4, (b) a HCDR2 of SEQ ID NO: 5, and (c) a HCDR3 of SEQ ID NO: 6, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 7, (e) a LCDR2 of SEQ ID NO: 8, and (f) a LCDR3 of SEQ ID NO: 9; or
d) a heavy chain variable region (VH) that comprises (a) a HCDR1 of SEQ ID NO: 14, (b) a HCDR2 of SEQ ID NO: 15, and (c) a HCDR3 of SEQ ID NO: 16, and a light chain variable region (VL) that comprises: (d) a LCDR1 of SEQ ID NO: 17, (e) a LCDR2 of SEQ ID NO: 18, and (f) a LCDR3 of SEQ ID NO: 19;
(ii). and the second antigen binding domain that specifically binds to human CD16A comprises:
a) a heavy chain variable region (VH) that comprises SEQ ID NO: 112;
b) a heavy chain variable region (VH) that comprises SEQ ID NO: 115;
c) a heavy chain variable region (VH) that comprises SEQ ID NO: 117; or
d) a heavy chain variable region (VH) that comprises SEQ ID NO: 119.
14 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein;
(i). the first antigen binding domain that specifically binds to human MUC1 comprises:
a) a heavy chain variable region (VH) that comprises SEQ ID NO: 30, and a light chain variable region (VL) that comprises SEQ ID NO: 31;
b) a heavy chain variable region (VH) that comprises SEQ ID NO: 61, and a light chain variable region (VL) that comprises SEQ ID NO: 62;
c) a heavy chain variable region (VH) that comprises SEQ ID NO: 61, and a light chain variable region (VL) that comprises SEQ ID NO: 66;
d) a heavy chain variable region (VH) that comprises SEQ ID NO: 61, and a light chain variable region (VL) that comprises SEQ ID NO: 68;
e) a heavy chain variable region (VH) that comprises SEQ ID NO: 10, and a light chain variable region (VL) that comprises SEQ ID NO: 11; or
f) a heavy chain variable region (VH) that comprises SEQ ID NO: 20, and a light chain variable region (VL) that comprises SEQ ID NO: 21;
(ii). and the second antigen binding domain that specifically binds to human CD16A comprises:
a) a heavy chain variable region (VH) that comprises SEQ ID NO: 112;
b) a heavy chain variable region (VH) that comprises SEQ ID NO: 115;
c) a heavy chain variable region (VH) that comprises SEQ ID NO: 117; or
d) a heavy chain variable region (VH) that comprises SEQ ID NO: 119.
15 . The multi-specific antibody or antigen-binding fragment of claim 1 , which is a monoclonal antibody, a chimeric antibody, a humanized antibody, a human engineered antibody, a single chain antibody (scFv), a Fab fragment, a Fab′ fragment, or a F(ab′)2 fragment.
16 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the multi-specific antibody is a bispecific antibody.
17 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the multi-specific antibody is BG1222P comprising SEQ ID NO: 143, SEQ ID NO: 145, and SEQ ID NO: 147.
18 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the antibody or antigen-binding fragment thereof has antibody dependent cellular cytotoxicity (ADCC) or complement dependent cytotoxicity (CDC).
19 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the antibody or antigen-binding fragment thereof has reduced glycosylation or no glycosylation or is hypofucosylated.
20 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the antibody or antigen-binding fragment thereof comprises increased bisecting GlcNac structures.
21 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the Fc domain is an IgG1 with reduced effector function.
22 . The multi-specific antibody or antigen-binding fragment of claim 1 , wherein the Fc domain is an IgG4.
23 . A pharmaceutical composition comprising the multi-specific antibody or antigen-binding fragment thereof of claim 1 and a pharmaceutically acceptable carrier.
24 . The pharmaceutical composition of claim 23 , comprising histidine/histidine HCl, trehalose dihydrate, and polysorbate 20.
25 . An isolated nucleic acid that encodes the multi-specific antibody or antigen-binding fragment of claim 1 .
26 . A vector comprising the nucleic acid of claim 25 .
27 . A host cell comprising the nucleic acid of claim 25 .
28 . A process for producing a multi-specific antibody or antigen-binding fragment thereof, comprising cultivating the host cell of claim 27 and recovering the antibody or antigen-binding fragment from the culture.
29 . A host cell comprising the vector of claim 26 .
30 . A method of treating cancer comprising administering to a patient in need an effective amount of the multi-specific antibody or antigen-binding fragment of claim 1 .
31 . The method of claim 25 , wherein the cancer is gastric cancer, colon cancer, pancreatic cancer, breast cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, ovarian cancer, skin cancer, mesothelioma, lymphoma, leukemia, myeloma and sarcoma.