Therapeutic cd47 antibodies

Abstract

Provided are monoclonal antibodies and antigen-binding fragments thereof that bind to CD47 of multiple mammalian species, block the binding of SIRPalpha and TSP1 to CD47, promote phagocytosis of susceptible cancer cells, and reverse TSP1 inhibition of nitric oxide signaling, as well as monoclonal antibodies and antigen binding fragments thereof that compete with the former for binding to CD47 and that exhibit similar biological activities. Also provided are combinations of any of the foregoing. Such antibody compounds are variously effective in 1) treating tissue ischemia and ischemia-reperfusion injury (IRI) in the setting of organ preservation and transplantation, pulmonary hypertension, sickle cell disease, myocardial infarction, stroke, and other instances of surgery and/or trauma in which IRI is a component of pathogenesis; 2) in treating autoimmune and inflammatory diseases; and 3) as anti-cancer agents for treating susceptible cancer cells, promoting their phagocytic uptake and clearance.

What is claimed is: 
     
         1 . A monoclonal antibody, or antigen-binding fragment thereof, that:
 (i) specifically binds human, rat, mouse, pig, cynomolgus monkey, and dog CD47;   (ii) blocks SIRPalpha and TSP1 binding to CD47;   (iii) promotes phagocytosis of cancer cells; and   (iv) reverses TSP1 inhibition of nitric oxide signaling.   
     
     
         2 . The monoclonal antibody or antigen-binding fragment thereof of  claim 1 , which is chimeric or humanized. 
     
     
         3 . The monoclonal antibody, or antigen-binding fragment thereof, of  claim 2 , which comprises three light chain complementarity determining regions (LCDRs 1-3) and three heavy chain complementarity determining regions (HCDRs 1-3), wherein:
 LCDR 1 comprises the amino acid sequence RSSQSLVHSNGNTYLH (SEQ ID NO:1) LCDR 2 comprises the amino acid sequence KVSYRFS (SEQ ID NO:2); and   LCDR 3 comprises the amino acid sequence SQNTHVPRT (SEQ ID NO:3);   HCDR1 comprises the amino acid sequence GYTFTNYYVF (SEQ ID NO:4);   HCDR 2 comprises the amino acid sequence DINPVNGDTNFNEKFKN (SEQ ID NO:5); and   HCDR 3 comprises the amino acid sequence GGYTMDY (SEQ ID NO:6).   
     
     
         4 . The monoclonal antibody, or antigen-binding fragment thereof of  claim 3 , which comprises a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein said LCVR and said HCVR comprise, respectively, amino acid sequences selected from among the following combinations of LCVRs and HCVRs:
 SEQ ID NO:7 and SEQ ID NO:57;   SEQ ID NO:8 and SEQ ID NO:58;   SEQ ID NO:9 and SEQ ID NO:59;   SEQ ID NO:10 and SEQ ID NO:60;   SEQ ID NO:11 and SEQ ID NO:61;   SEQ ID NO:12 and SEQ ID NO:62;   SEQ ID NO:13 and SEQ ID NO:63;   SEQ ID NO:14 and SEQ ID NO:64;   SEQ ID NO:15 and SEQ ID NO:65;   SEQ ID NO:16 and SEQ ID NO:66;   SEQ ID NO:17 and SEQ ID NO:67;   SEQ ID NO:18 and SEQ ID NO:68;   SEQ ID NO:19 and SEQ ID NO:69;   SEQ ID NO:20 and SEQ ID NO:70;   SEQ ID NO:21 and SEQ ID NO:71;   SEQ ID NO:22 and SEQ ID NO:72;   SEQ ID NO:23 and SEQ ID NO:73;   SEQ ID NO:24 and SEQ ID NO:74;   SEQ ID NO:25 and SEQ ID NO:75;   SEQ ID NO:26 and SEQ ID NO:76;   SEQ ID NO:27 and SEQ ID NO:77;   SEQ ID NO:28 and SEQ ID NO:78;   SEQ ID NO:29 and SEQ ID NO:79;   SEQ ID NO:30 and SEQ ID NO:80; and   SEQ ID NO:31 and SEQ ID NO:81,   wherein each one of LCVR SEQ ID NOs:7-31 further comprises a constant domain having the amino acid sequence shown in SEQ ID NO:117,   and   wherein each one of HCVR SEQ ID NOs:57-81 comprises a constant domain selected from among SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, and SEQ ID NO:121.   
     
     
         5 . A monoclonal antibody, or antigen-binding fragment thereof, that competes with said monoclonal antibody or antigen binding fragment thereof of  claim 4  for binding to human CD47. 
     
     
         6 . A pharmaceutical composition, comprising said monoclonal antibody or antigen-binding fragment thereof, or competing antibody or antigen binding fragment thereof, of  claim 1  and a pharmaceutically or physiologically acceptable carrier, diluent, or excipient. 
     
     
         7 . A monoclonal antibody or antigen-binding fragment thereof, or competing antibody or antigen binding fragment thereof, of  claim 1  for use in human therapy. 
     
     
         8 . The monoclonal antibody or antigen-binding fragment thereof, or competing antibody or antigen binding fragment thereof of  claim 1  for use in reducing, preventing, and/or treating ischemia-reperfusion injury, or an autoimmune or inflammatory disease in a human patient. 
     
     
         9 . The monoclonal antibody or antigen-binding fragment thereof, or competing antibody or antigen binding fragment thereof, of  claim 8 , wherein said ischemia-reperfusion injury occurs in organ transplantation, acute kidney injury, cardiopulmonary bypass surgery, pulmonary hypertension, sickle cell disease, myocardial infarction, stroke, surgical resections and reconstructive surgery, reattachment of an appendage or other body part, skin grafting, or trauma. 
     
     
         10 . The monoclonal antibody, or antigen-binding fragment thereof, or competing antibody or antigen binding fragment thereof, of  claim 8 , wherein said autoimmune or inflammatory disease is selected from the group consisting of arthritis, multiple sclerosis, psoriasis, Crohn's disease, inflammatory bowel disease, lupus, Grave's disease and Hashimoto's thyroiditis, and ankylosing spondylitis. 
     
     
         11 . The monoclonal antibody or antigen-binding fragment thereof, or competing antibody or antigen binding fragment thereof of  claim 1  for use in treating or reducing a susceptible cancer. 
     
     
         12 . The monoclonal antibody or antigen binding fragment thereof, or competing antibody or antigen binding fragment thereof, of  claim 11 , which promotes phagocytosis of cells of said susceptible cancer. 
     
     
         13 . The monoclonal antibody or antigen binding fragment thereof, or competing antibody or antigen binding fragment thereof, of  claim 11  wherein said susceptible cancer is selected from the group consisting of a leukemia, a lymphoma, ovarian cancer, breast cancer, endometrial cancer, colon cancer, rectal cancer, gastric cancer, bladder cancer, lung cancer, bronchial cancer, bone cancer, prostate cancer, pancreatic cancer, liver and bile duct cancer, esophageal cancer, renal cancer, thyroid cancer, head and neck cancer, testicular cancer, glioblastoma, astrocytoma, melanoma, myelodysplastic syndrome, and a sarcoma. 
     
     
         14 . The monoclonal antibody or antigen binding fragment thereof, or competing antibody or antigen binding fragment thereof, of  claim 11  wherein:
 said leukemia is selected from the group consisting of acute lymphocytic (lymphoblastic) leukemia, acute myeloid leukemia, myelogenous leukemia, chronic lymphocytic leukemia, multiple myeloma, plasma cell leukemia, and chronic myeloid leukemia; 
 said lymphoma is selected from the group consisting of Hodgkin lymphoma and Non-Hodgkin lymphoma including B cell lymphoma, diffuse large B cell lymphoma, follicular lymphoma, mantle cell lymphoma, marginal zone B cell lymphoma, T cell lymphoma, and Waldenstrom macroglobulinemia; and 
 said sarcoma is selected from the group consisting of osteosarcoma, Ewing sarcoma, leiomyosarcoma, synovial sarcoma, alveolar soft part sarcoma, angiosarcoma, liposarcoma, fibrosarcoma, rhabdomyosarcoma, and chrondrosarcoma. 
 
     
     
         15 . Use of said monoclonal antibody or antigen-binding fragment thereof, or competing antibody or antigen binding fragment thereof, of  claim 1  to reduce, prevent, and/or treat ischemia-reperfusion injury, or an autoimmune or inflammatory disease, in a human patient. 
     
     
         16 . Use of said monoclonal antibody or antigen-binding fragment thereof, or competing antibody or antigen binding fragment thereof, of  claim 1  to reduce, prevent, and/or treat a susceptible cancer. 
     
     
         17 . Use of said monoclonal antibody or antigen-binding fragment thereof, or competing antibody or antigen binding fragment thereof, of  claim 1  for the manufacture of a medicament to prevent, reduce, and/or treat ischemia-reperfusion injury, or an autoimmune or inflammatory disease, in a human patient. 
     
     
         18 . The use of  claim 17 , wherein said ischemia-reperfusion injury occurs in organ transplantation, acute kidney injury, cardiopulmonary bypass surgery, pulmonary hypertension, sickle cell disease, myocardial infarction, stroke, surgical resections and reconstructive surgery, reattachment of an appendage or other body part, skin grafting, and trauma. 
     
     
         19 . The use of  claim 17 , wherein said autoimmune or inflammatory disease is selected from among arthritis, multiple sclerosis, psoriasis, Crohn's disease, inflammatory bowel disease, lupus, Grave's disease and Hashimoto's thyroiditis, and ankylosing spondylitis. 
     
     
         20 . Use of said monoclonal antibody or antigen-binding fragment thereof, or competing antibody or antigen binding fragment thereof, of  claim 1  for the manufacture of a medicament to treat or reduce a susceptible cancer. 
     
     
         21 . A monoclonal antibody, or antigen-binding fragment thereof, of  claim 1  that specifically binds to epitopes in the extracellular IgV domain of CD47. 
     
     
         22 . A monoclonal antibody, or antigen-binding fragment thereof, of  claim 5  that specifically binds to epitopes in the extracellular IgV domain of CD47.