Combination therapy of cancer involving multi-specific binding proteins that bind nkg2d, cd16, and a tumor-associated antigen

Abstract

Combination therapy of a cancer with a multi-specific binding protein that bind a tumor associated antigen, the NKG2D receptor, and CD16, in combination with a second anti-cancer agent are described. Also described are pharmaceutical compositions of the multi-specific binding protein, and therapeutic methods useful for the treatment of cancer in combination with a second anti-cancer agent.

1 . A method of enhancing cell death, directly or indirectly, of a tumor cell expressing a tumor-associated antigen, the method comprising exposing the tumor cell and a natural killer cell to a multi-specific binding protein comprising:
 (a) a first antigen-binding site that binds NKG2D;   (b) a second antigen-binding site that binds the tumor-associated antigen; and   (c) an antibody Fc domain or a portion thereof sufficient to bind CD16, or a third antigen-binding site that binds CD16; 
in combination with a second therapeutic agent comprising an anti-LAG3 antibody. 
     
     
         2 . A method of treating a cancer expressing a tumor-associated antigen in a subject in need thereof, the method comprising administering to the subject effective amounts of a multi-specific binding protein comprising:
 (a) a first antigen-binding site that binds NKG2D;   (b) a second antigen-binding site that binds the tumor-associated antigen; and   (c) an antibody Fc domain or a portion thereof sufficient to bind CD16, or a third antigen-binding site that binds CD16;   and a second therapeutic agent comprising an anti-LAG3 antibody.   
     
     
         3 - 32 . (canceled) 
     
     
         33 . The method of  claim 2 , wherein the cancer is selected from the group consisting of acute myeloid leukemia, acute myelomonocytic leukemia, B cell lymphoma, bladder cancer, breast cancer, colorectal cancer, diffuse large B cell lymphoma esophageal cancer, Ewing’s sarcoma, follicular lymphoma, gastric cancer, gastrointestinal cancer, gastrointestinal stromal tumors, glioblastoma, head and neck cancer, melanoma, mesothelioma, multiple myeloma, myelodysplastic syndrome, renal cell carcinoma, neuroblastoma, non-small cell lung cancer, neuroendocrine tumors, ovarian cancer, and pancreatic cancer, prostate cancer, sarcomas, small cell lung cancer, T cell lymphoma, testis cancer, thymic carcinoma, thyroid cancer, urothelial cancer, cancers infiltrated by myeloid-derived suppressor cells, cancers with extracellular matrix deposition, cancers with high levels of reactive stroma, and cancers with neoangiogenesis. 
     
     
         34 . The method of  claim 2 , wherein the first antigen-binding site of the multi-specific binding protein binds to NKG2D in humans and non-human primates. 
     
     
         35 . The method of  claim 2 , wherein the first antigen-binding site comprises a heavy chain variable domain and a light chain variable domain. 
     
     
         36 - 42 . (canceled) 
     
     
         43 . The method of any one of  claim 2 , wherein the second antigen-binding site comprises a heavy chain variable domain and a light chain variable domain. 
     
     
         44 . The method of  claim 2 , wherein the tumor-associated antigen is selected from the group consisting of HER2, EpCAM, CD2, CD20, CD30, CD38, CD40, CD52, CD70, EGFR/ERBB1, IGF1R, HER3/ERBB3, HER4/ERBB4, MUC1, SLAMF7, PSCA, MICA, MICB, TRAILR1, TRAILR2, MAGE-A3, B7.1, B7.2, CTLA4, and PD-L1. 
     
     
         45 . The method of any one of  claim 2 , wherein the multi-specific binding protein comprises a portion of an antibody Fc domain sufficient to bind CD16, wherein the antibody Fc domain comprises hinge and CH2 domains. 
     
     
         46 . The method of  claim 2 , wherein the multi-specific binding protein comprises an amino acid sequence at least 90% identical to amino acids 234-332 of a human IgG1 antibody. 
     
     
         47 . The method of  claim 2 , wherein the multi-specific binding is administered in a formulation that further comprises a pharmaceutically acceptable carrier. 
     
     
         48 . The method of  claim 2 , wherein the first antigen binding site comprises a heavy chain variable domain comprising a complementarity-determining region (CDR) 1 amino acid sequence of SEQ ID NO:92, a CDR2 amino acid sequence of SEQ ID NO:58, and a CDR3 amino acid sequence of SEQ ID NO: 113; and a light chain variable domain comprising a CDR1 amino acid sequence of SEQ ID NO:60, a CDR2 amino acid sequence of SEQ ID NO:61, and a CDR3 amino acid sequence of SEQ ID NO:62. 
     
     
         49 . The method of  claim 48 , wherein the first antigen-binding site comprises a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:47 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:48. 
     
     
         50 . The method of  claim 2 , wherein the first antigen binding site comprises a heavy chain variable domain comprising a CDR1 amino acid sequence of SEQ ID NO:57, a CDR2 amino acid sequence of SEQ ID NO:58, and a CDR3 amino acid sequence of SEQ ID NO:59; and a light chain variable domain comprising a CDR1 amino acid sequence of SEQ ID NO:60, a CDR2 amino acid sequence of SEQ ID NO:61, and a CDR3 amino acid sequence of SEQ ID NO:62. 
     
     
         51 . The method of  claim 50 , wherein the first antigen-binding site comprises a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:47 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:48. 
     
     
         52 . The method of  claim 2 , wherein the first antigen binding site comprises:
 (a) a heavy chain variable domain comprising a CDR1 amino acid sequence of SEQ ID NO:90, a CDR2 amino acid sequence of SEQ ID NO:52, and a CDR3 amino acid sequence of SEQ ID NO:91; and a light chain variable domain comprising a CDR1 amino acid sequence of SEQ ID NO:54, a CDR2 amino acid sequence of SEQ ID NO:55, and a CDR3 amino acid sequence of SEQ ID NO:56;   (b) a heavy chain variable domain comprising a CDR1 amino acid sequence of SEQ ID NO:94, a CDR2 amino acid sequence of SEQ ID NO:64, and a CDR3 amino acid sequence of SEQ ID NO:95; and a light chain variable domain comprising a CDR1 amino acid sequence of SEQ ID NO:66, a CDR2 amino acid sequence of SEQ ID NO:67, and a CDR3 amino acid sequence of SEQ ID NO:68;   (c) a heavy chain variable domain comprising a CDR1 amino acid sequence of SEQ ID NO:122, a CDR2 amino acid sequence of SEQ ID NO:117, and a CDR3 amino acid sequence of SEQ ID NO:123; and a light chain variable domain comprising a CDR1 amino acid sequence of SEQ ID NO:119, a CDR2 amino acid sequence of SEQ ID NO:120, and a CDR3 acid sequence of SEQ ID NO:121; or   (d) a heavy chain variable domain comprising a CDR1 amino acid sequence of SEQ ID NO:122, a CDR2 amino acid sequence of SEQ ID NO:117, and a CDR3 amino acid sequence of SEQ ID NO:130; and a light chain variable domain comprising a CDR1 amino acid sequence of SEQ ID NO:127, a CDR2 amino acid sequence of SEQ ID NO:128, and a CDR3 amino acid sequence of SEQ ID NO:129.   
     
     
         53 . The method of  claim 52 , wherein the first antigen-binding site comprises:
 (a) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:45 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:46, wherein the heavy chain variable domain comprises a CDR1 amino acid sequence of SEQ ID NO:90, a CDR2 amino acid sequence of SEQ ID NO:52, and a CDR3 amino acid sequence of SEQ ID NO:91; and the light chain variable domain comprises a CDR1 amino acid sequence of SEQ ID NO:54, a CDR2 amino acid sequence of SEQ ID NO:55, and a CDR3 amino acid sequence of SEQ ID NO:56;   (b) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:49 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:50, wherein the heavy chain variable domain comprises a CDR1 amino acid sequence of SEQ ID NO:94, a CDR2 amino acid sequence of SEQ ID NO:64, and a CDR3 amino acid sequence of SEQ ID NO:95; and the light chain variable domain comprises a CDR1 amino acid sequence of SEQ ID NO:66, a CDR2 amino acid sequence of SEQ ID NO:67, and a CDR3 amino acid sequence of SEQ ID NO:68;   (c) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:114 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:115, wherein the heavy chain variable domain comprises a CDR1 amino acid sequence of SEQ ID NO:122, a CDR2 amino acid sequence of SEQ ID NO:117, and a CDR3 amino acid sequence of SEQ ID NO:123; and the light chain variable domain comprises a CDR1 amino acid sequence of SEQ ID NO:119, a CDR2 amino acid sequence of SEQ ID NO:120, and a CDR3 acid sequence of SEQ ID NO:121; or   (d) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:124 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:125, wherein the heavy chain variable domain comprises a CDR1 amino acid sequence of SEQ ID NO:122, a CDR2 amino acid sequence of SEQ ID NO:117, and a CDR3 amino acid sequence of SEQ ID NO:130; and the light chain variable domain comprises a CDR1 amino acid sequence of SEQ ID NO:127, a CDR2 amino acid sequence of SEQ ID NO:128, and a CDR3 amino acid sequence of SEQ ID NO:129.   
     
     
         54 . The method of  claim 33 , wherein the cancer is breast cancer.