PROTEINS BINDING BCMA, NKG2D AND CD16
Multi-specific binding proteins that bind BCMA, the NKG2D receptor, and CD 16 are described, as well as pharmaceutical compositions and therapeutic methods useful for the treatment of cancer.
1 . A protein comprising:
(a) a first antigen-binding site that binds NKG2D;
(b) a second antigen-binding site that binds BCMA; and
(c) an antibody Fc domain or a portion thereof sufficient to bind CD16, or a third antigen-binding site that binds CD16.
2 . The protein of claim 1 , wherein the first antigen-binding site binds to NKG2D in humans, non-human primates, and rodents.
3 . The protein of claim 1 or 2 , wherein the first antigen-binding site comprises a heavy chain variable domain and a light chain variable domain.
4 . A protein according to claim 3 , wherein the heavy chain variable domain and the light chain variable domain are present on the same polypeptide.
5 . A protein according to any one of claims 3 - 4 , wherein the second antigen-binding site comprises a heavy chain variable domain and a light chain variable domain.
6 . A protein according to claim 5 , wherein the heavy chain variable domain and the light chain variable domain of the second antigen-binding site are present on the same polypeptide.
7 . A protein according to claim 5 or 6 , wherein the light chain variable domain of the first antigen-binding site has an amino acid sequence identical to the amino acid sequence of the light chain variable domain of the second antigen-binding site.
8 . A protein according to any one of the preceding claims, wherein the first antigen-binding site comprises a heavy chain variable domain at least 90% identical to SEQ ID NO: 1.
9 . A protein according to any of claims 1 - 7 , wherein the first antigen-binding site comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:41 and a light chain variable domain at least 90% identical to SEQ ID NO:42.
10 . A protein according to any of claims 1 - 7 , wherein the first antigen-binding site comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:43 and a light chain variable domain at least 90% identical to SEQ ID NO:44.
11 . A protein according to any of claims 1 - 7 , wherein the first antigen-binding site comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:45 and a light chain variable domain at least 90% identical to SEQ ID NO:46.
12 . A protein according to any of claims 1 - 7 , wherein the first antigen-binding site comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:47 and a light chain variable domain at least 90% identical to SEQ ID NO:48.
13 . The protein of claim 1 or 2 , wherein the first antigen-binding site is a single-domain antibody.
14 . The protein of claim 13 , wherein the single-domain antibody is a V H H fragment or a V NAR fragment.
15 . A protein according to any one of claims 1 - 2 or 13 - 14 , wherein the second antigen-binding site comprises a heavy chain variable domain and a light chain variable domain.
16 . A protein according to claim 15 , wherein the heavy chain variable domain and the light chain variable domain of the second antigen-binding site are present on the same polypeptide.
17 . A protein according to any of the preceding claims, wherein the heavy chain variable domain of the second antigen-binding site comprises an amino acid sequence at least 90% identical to SEQ ID NO:49 and the light chain variable domain of the second antigen-binding site comprises an amino acid sequence at least 90% identical to SEQ ID NO:53 or SEQ ID NO:54.
18 . A protein according to any of the preceding claims, wherein the heavy chain variable domain of the second antigen-binding site comprises an amino acid sequence including:
a heavy chain CDR1 sequence identical to the amino acid sequence of SEQ ID NO:50;
a heavy chain CDR2 sequence identical to the amino acid sequence of SEQ ID NO:51; and
a heavy chain CDR3 sequence identical to the amino acid sequence of SEQ ID NO:52.
19 . A protein according to claim 18 , wherein the light chain variable domain of the second antigen-binding site comprises an amino acid sequence including:
a light chain CDR1 sequence identical to the amino acid sequence of SEQ ID NO:55;
a light chain CDR2 sequence identical to the amino acid sequence of SEQ ID NO:56; and
a light chain CDR3 sequence identical to the amino acid sequence of SEQ ID NO:57 or SEQ ID NO:57.
20 . A protein according to any one of claims 1 - 16 , wherein the heavy chain variable domain of the second antigen-binding site comprises an amino acid sequence at least 90% identical to SEQ ID NO:59 and the light chain variable domain of the second antigen-binding site comprises an amino acid sequence at least 90% identical to SEQ ID NO:60.
21 . A protein according to any one of claims 1 - 16 or 20 , wherein the heavy chain variable domain of the second antigen-binding site comprises an amino acid sequence including:
a heavy chain CDR1 sequence identical to the amino acid sequence of SEQ ID NO:79;
a heavy chain CDR2 sequence identical to the amino acid sequence of SEQ ID NO:80; and
a heavy chain CDR3 sequence identical to the amino acid sequence of SEQ ID NO:81.
22 . A protein according to claim 21 , wherein the light chain variable domain of the second antigen-binding site comprises an amino acid sequence including:
a light chain CDR1 sequence identical to the amino acid sequence of SEQ ID NO:82;
a light chain CDR2 sequence identical to the amino acid sequence of SEQ ID NO:83; and
a light chain CDR3 sequence identical to the amino acid sequence of SEQ ID NO:84.
23 . A protein according to any one of claims 1 - 16 , wherein the heavy chain variable domain of the second antigen-binding site comprises an amino acid sequence at least 90% identical to SEQ ID NO:61 and the light chain variable domain of the second antigen-binding site comprises an amino acid sequence at least 90% identical to SEQ ID NO:62.
24 . A protein according to any one of claims 1 - 16 or 23 , wherein the heavy chain variable domain of the second antigen-binding site comprises an amino acid sequence including:
a heavy chain CDR1 sequence identical to the amino acid sequence of SEQ ID NO:85;
a heavy chain CDR2 sequence identical to the amino acid sequence of SEQ ID NO:86; and
a heavy chain CDR3 sequence identical to the amino acid sequence of SEQ ID NO:87.
25 . A protein according to any one of claim 24 , wherein the light chain variable domain of the second antigen-binding site comprises an amino acid sequence including:
a light chain CDR1 sequence identical to the amino acid sequence of SEQ ID NO:88;
a light chain CDR2 sequence identical to the amino acid sequence of SEQ ID NO:89; and
a light chain CDR3 sequence identical to the amino acid sequence of SEQ ID NO:90.
26 . A protein according to any one of claims 1 - 4 or 8 - 14 , wherein the second antigen-binding site is a single-domain antibody.
27 . The protein of claim 26 , wherein the second antigen-binding site is a V H H fragment or a V NAR fragment.
28 . A protein according to any one of the preceding claims, wherein the protein comprises a portion of an antibody Fc domain sufficient to bind CD16, wherein the antibody Fc domain comprises hinge and CH2 domains.
29 . A protein according to claim 28 , wherein the antibody Fc domain comprises hinge and CH2 domains of a human IgG1 antibody.
30 . A protein according to claim 28 or 29 , wherein the Fc domain comprises an amino acid sequence at least 90% identical to amino acids 234-332 of a human IgG1 antibody.
31 . A protein according to any one of claims 28 - 30 , wherein the Fc domain comprises amino acid sequence at least 90% identical to the Fc domain of human IgG1 and differs at one or more positions selected from the group consisting of Q347, Y349, L351, S354, E356, E357, K360, Q362, S364, T366, L368, K370, N390, K392, T394, D399, S400, D401, F405, Y407, K409, T411, K439.
32 . A formulation comprising a protein according to any one of the preceding claims and a pharmaceutically acceptable carrier.
33 . A cell comprising one or more nucleic acids expressing a protein according to any one of claims 1 - 31 .
34 . A method of directly and/or indirectly enhancing tumor cell death, the method comprising exposing a tumor and natural killer cells to a protein according to any one of claims 1 - 31 .
35 . A method of treating cancer, wherein the method comprises administering a protein according to any one of claims 1 - 31 or a formulation according to claim 32 to a patient.
36 . The method of claim 35 , wherein the cancer is selected from the group consisting of multiple myeloma, acute myelomonocytic leukemia, T cell lymphoma, acute monocytic leukemia, and follicular lymphoma.