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 
                 
                 
               
                   (SEQ ID NO: 1) 
                 
                 
                 
               
                     
                   RSSQSLVHSNGNTYLH 
                 
                     
                 
                     
                   LCDR 2 comprises the amino acid sequence 
                 
                 
               
                   (SEQ ID NO: 2) 
                 
                 
                 
               
                     
                   KVSYRFS; 
                 
                     
                   and 
                 
                     
                 
                     
                   LCDR 3 comprises the amino acid sequence 
                 
                 
               
                   (SEQ ID NO: 3) 
                 
                 
                 
               
                     
                   SQNTHVPRT; 
                 
                     
                 
                     
                   HCDR1 comprises the amino acid sequence 
                 
                 
               
                   (SEQ ID NO: 4) 
                 
                 
                 
               
                     
                   GYTFTNYYVF; 
                 
                     
                 
                     
                   HCDR 2 comprises the amino acid sequence 
                 
                 
               
                   (SEQ ID NO: 5) 
                 
                 
                 
               
                     
                   DINPVNGDTNFNEKFKN; 
                 
                     
                   and 
                 
                     
                 
                     
                   HCDR 3 comprises the amino acid sequence 
                 
                 
               
                   (SEQ ID NO: 6) 
                 
                 
                 
               
                     
                   GGYTMDY. 
                 
             
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
                
               
            
             
                
               
            
             
                
               
            
           
         
       
     
     
         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, SEQ ID NO:121, and SEQ ID NO:124. 
     
     
         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 (colorectal cancer), rectal cancer, bladder cancer, lung cancer (non-small cell lung cancer, adenocarcinoma of the lung, squamous cell carcinoma of the lung), bronchial cancer, bone cancer, prostate cancer, pancreatic cancer, gastric cancer, hepatocellular carcinoma, gall bladder cancer, bile duct cancer, esophageal cancer, renal cell carcinoma, thyroid cancer, squamous cell carcinoma of the head and neck (head and neck cancer), testicular cancer, cancer of the endocrine gland, cancer of the adrenal gland, cancer of the pituitary gland, cancer of the skin, cancer of soft tissues, cancer of blood vessels, cancer of brain, cancer of nerves, cancer of eyes, cancer of meninges, cancer of oropharynx, cancer of hypopharynx, cancer of cervix, and cancer of uterus, glioblastoma, meduloblastoma, astrocytoma, glioma, meningioma, gastrinoma, neuroblastoma, 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 13  wherein:
 said leukemia is selected from the group consisting of systemic mastocytosis, acute lymphocytic (lymphoblastic) leukemia (ALL), T cell—ALL, acute myeloid leukemia (AML), myelogenous leukemia, chronic lymphocytic leukemia (CLL), multiple myeloma (MM), chronic myeloid leukemia (CML), myeloproliferative disorder/neoplasm, myelodysplastic syndrome, monocytic cell leukemia, and plasma cell leukemia; 
 wherein said lymphoma is selected from the group consisting of histiocytic lymphoma and T cell lymphoma, B cell lymphomas, including Hodgkin's lymphoma and non-Hodgkin's lymphoma, such as low grade/follicular non-Hodgkin's lymphoma (NHL), cell lymphoma (FCC), mantle cell lymphoma (MCL), diffuse large cell lymphoma (DLCL), small lymphocytic (SL) NHL, intermediate grade/follicular NHL, intermediate grade diffuse NHL, high grade immunoblastic NHL, high grade lymphoblastic NHL, high grade small non-cleaved cell NHL, bulky disease NHL, and Waldenstrom's Macroglobulinemia; and 
 wherein 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.