Therapeutic cd47 antibodies

Abstract

Provided are anti-CD47 monoclonal antibodies (anti-CD47 mAbs) with distinct functional profiles as described herein, methods to generate anti-CD47 mAbs, and to methods of using these anti-CD47 mAbs as therapeutics for the prevention and treatment of solid and hematological cancers, ischemia-reperfusion injury, cardiovascular diseases, autoimmune diseases, inflammatory diseases or as diagnostics for determining the level of CD47 in tissue samples.

1 - 101 . (canceled) 
     
     
         102 . A monoclonal antibody, or an antigen binding fragment thereof, which:
 a. binds to human CD47;   b. blocks SIRP (binding to human CD47;   c. increases phagocytosis of human tumor cells;   d. induces death of human tumor cells;   e. exhibits pH-dependent binding to human CD47 present on a cell; and
 wherein the monoclonal antibody, or an antigen binding fragment thereof, further comprises one or more characteristics selected from the group consisting of: 
 i) causes no detectable agglutination of human red blood cells, or causes reduced agglutination of human red blood cells, and 
 ii) has reduced human red blood cell binding, or has minimal human red blood cell binding. 
   
     
     
         103 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 102 , wherein said monoclonal antibody, or an antigen binding fragment thereof, possesses one or more among the following characteristics:
 causes an increase in cell surface calreticulin expression on human tumor cells;   causes an increase in adenosine triphosphate (ATP) release by human tumor cells;   causes an increase in high mobility group box 1 (HMGB1) release by human tumor cells;   causes an increase in annexin A1 release by human tumor cells;   causes an increase in Type I Interferon release by human tumor cells;   causes an increase in C-X-C Motif Chemokine Ligand 10 (CXCL10) release by human tumor cells;   causes an increase in cell surface protein disulfide-isomerase A3 (PDIA3) expression on human tumor cells;   causes an increase in cell surface heat shock protein 70 (HSP70) expression on human tumor cells; and   causes an increase in cell surface heat shock protein 90 (HSP90) expression on human tumor cells.   
     
     
         104 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 102 , wherein the antibody, or antigen binding fragment thereof, is a chimeric or humanized antibody;
 or wherein the monoclonal antibody, or antigen binding fragment thereof cross-reacts with one or more species homologs of CD47.   
     
     
         105 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 102 , wherein the antibody, or antigen binding fragment thereof, causes no detectable agglutination of human red blood cells and has minimal human red blood cell binding. 
     
     
         106 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 102 , wherein the pH-dependent binding to human CD47 has a greater affinity for human CD47 at an acidic pH compared to physiological pH. 
     
     
         107 . A method of treating a disease comprising an autoimmune disease, an autoinflammatory disease, an inflammatory disease, a cardiovascular disease, or a cancer in a subject, comprising administration of the monoclonal antibody or antigen-binding fragment thereof, of  claim 102 , wherein the subject comprises a human or companion/pet animal, a working animal, a sport animal, a zoo animal, or a valuable animal kept in captivity. 
     
     
         108 . The method of  claim 107 , wherein the disease 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, urothelial 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, a sarcoma, ischemia-reperfusion injury, heart failure, arthritis, rheumatoid arthritis, multiple sclerosis, psoriasis, psoriatic arthritis, Crohn's disease, inflammatory bowel disease, ulcerative colitis, lupus, systemic lupus erythematous, juvenile rheumatoid arthritis, juvenile idiopathic arthritis, Grave's disease, Hashimoto's thyroiditis, Addison's disease, celiac disease, dermatomyositis, multiple sclerosis, myasthenia gravis, pernicious anemia, Sjogren syndrome, type I diabetes, vasculitis, uveitis, atherosclerosis, and ankylosing spondylitis. 
     
     
         109 . A monoclonal antibody, or an antigen binding fragment thereof, which:
 a. binds to human CD47;   b. blocks SIRPα binding to human CD47;   c. increases phagocytosis of human tumor cells;   d. induces death of human tumor cells;   e. exhibits pH-dependent binding to human CD47 present on a cell;   f. exhibits reduced binding to a normal cell; and
 wherein the monoclonal antibody, or an antigen binding fragment thereof, further comprises one or more characteristics selected from the group consisting of: 
 i) causes no detectable agglutination of human red blood cells, or causes reduced agglutination of human red blood cells; and 
 ii) has reduced human red blood cell binding, or has minimal human red blood cell binding. 
   
     
     
         110 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 109 , wherein said monoclonal antibody, or an antigen binding fragment thereof, further comprises one or more characteristics selected from the group consisting of:
 causes an increase in cell surface calreticulin expression on human tumor cells;   causes an increase in adenosine triphosphate (ATP) release by human tumor cells;   causes an increase in high mobility group box 1 (HMGB1) release by human tumor cells;   causes an increase in annexin A1 release by human tumor cells;   causes an increase in Type I Interferon release by human tumor cells;   causes an increase in C-X-C Motif Chemokine Ligand 10 (CXCL10) release by human tumor cells;   causes an increase in cell surface protein disulfide-isomerase A3 (PDIA3) expression on human tumor cells;   causes an increase in cell surface heat shock protein 70 (HSP70) expression on human tumor cells; and   causes an increase in cell surface heat shock protein 90 (HSP90) expression on human tumor cells.   
     
     
         111 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 109 , wherein the antibody, or antigen binding fragment thereof, is a chimeric or humanized antibody; or wherein the monoclonal antibody, or antigen binding fragment thereof cross-reacts with one or more species homologs of CD47. 
     
     
         112 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 109 , wherein the antibody, or antigen binding fragment thereof, causes no detectable agglutination of human red blood cells and has minimal human red blood cell binding. 
     
     
         113 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 109 , wherein the pH-dependent binding to CD47 has a greater affinity for CD47 at an acidic pH compared to physiological pH. 
     
     
         114 . A method of treating a disease comprising an autoimmune disease, an autoinflammatory disease, an inflammatory disease, a cardiovascular disease, or a cancer in a subject, comprising administration of the monoclonal antibody or antigen-binding fragment thereof, of  claim 109 , wherein subject comprises a human or companion/pet animal, a working animal, a sport animal, a zoo animal, or a valuable animal kept in captivity. 
     
     
         115 . The method of  claim 114 , wherein the disease 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, urothelial 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, a sarcoma, ischemia-reperfusion injury, heart failure, arthritis, rheumatoid arthritis, multiple sclerosis, psoriasis, psoriatic arthritis, Crohn's disease, inflammatory bowel disease, ulcerative colitis, lupus, systemic lupus erythematous, juvenile rheumatoid arthritis, juvenile idiopathic arthritis, Grave's disease, Hashimoto's thyroiditis, Addison's disease, celiac disease, dermatomyositis, multiple sclerosis, myasthenia gravis, pernicious anemia, Sjogren syndrome, type I diabetes, vasculitis, uveitis, atherosclerosis, and ankylosing spondylitis. 
     
     
         116 . A monoclonal antibody, or an antigen binding fragment thereof, which binds to human CD47, wherein said monoclonal antibody, or an antigen binding fragment thereof, exhibits pH-dependent binding to CD47 present on a cell. 
     
     
         117 . The monoclonal antibody, or an antigen binding fragment thereof, of  claim 116 , which:
 a. binds to human CD47;   b. blocks SIRPα binding to human CD47;   c. increases phagocytosis of human tumor cells; and   d. induces death of human tumor cells.   
     
     
         118 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 116 , wherein said monoclonal antibody, or an antigen binding fragment thereof, further comprises one or more characteristics selected from the group consisting of:
 causes an increase in cell surface calreticulin expression on human tumor cells;   causes an increase in adenosine triphosphate (ATP) release by human tumor cells;   causes an increase in high mobility group box 1 (HMGB1) release by human tumor cells;   causes an increase in annexin A1 release by human tumor cells;   causes an increase in Type I Interferon release by human tumor cells;   causes an increase in C-X-C Motif Chemokine Ligand 10 (CXCL10) release by human tumor cells;   causes an increase in cell surface protein disulfide-isomerase A3 (PDIA3) expression on human tumor cells;   causes an increase in cell surface heat shock protein 70 (HSP70) expression on human tumor cells; and   causes an increase in cell surface heat shock protein 90 (HSP90) expression on human tumor cells.   
     
     
         119 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 116 , wherein the antibody, or antigen binding fragment thereof, is a chimeric or humanized antibody; or
 wherein the monoclonal antibody, or antigen binding fragment thereof cross-reacts with one or more species homologs of CD47.   
     
     
         120 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 116 , wherein the pH-dependent binding to CD47 has a greater affinity for CD47 at an acidic pH compared to physiological pH. 
     
     
         121 . A method of treating a disease comprising an autoimmune disease, an autoinflammatory disease, an inflammatory disease, a cardiovascular disease, or a cancer in a subject, comprising administration of the monoclonal antibody or antigen-binding fragment thereof, of  claim 116 , wherein the subject comprises a human or companion/pet animal, a working animal, a sport animal, a zoo animal, or a valuable animal kept in captivity. 
     
     
         122 . The method of  claim 121 , wherein the disease 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, urothelial 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, a sarcoma, ischemia-reperfusion injury, heart failure, arthritis, rheumatoid arthritis, multiple sclerosis, psoriasis, psoriatic arthritis, Crohn's disease, inflammatory bowel disease, ulcerative colitis, lupus, systemic lupus erythematous, juvenile rheumatoid arthritis, juvenile idiopathic arthritis, Grave's disease, Hashimoto's thyroiditis, Addison's disease, celiac disease, dermatomyositis, multiple sclerosis, myasthenia gravis, pernicious anemia, Sjogren syndrome, type I diabetes, vasculitis, uveitis, atherosclerosis, and ankylosing spondylitis. 
     
     
         123 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 116 , wherein the monoclonal antibody, or antigen binding fragment thereof, further exhibits reduced binding to a normal cell. 
     
     
         124 . A monoclonal antibody, or an antigen binding fragment thereof, which binds to human CD47, wherein said monoclonal antibody, or an antigen binding fragment thereof, exhibits reduced binding to a normal cell. 
     
     
         125 . The monoclonal antibody, or an antigen binding fragment thereof, of  claim 124 , which:
 a. binds to human CD47;   b. blocks SIRPα binding to human CD47;   c. increases phagocytosis of human tumor cells; and   d. induces death of human tumor cells.   
     
     
         126 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 124 , wherein the normal cell is selected from the group consisting of an endothelial cell, a skeletal muscle cell, an epithelial cell, a PBMC, a T cell, a red blood cell, a peripheral blood mononuclear cell, a human aortic endothelial cell, a human skeletal muscle cell, a human microvascular endothelial cell, a human renal tubular epithelial cell, a human peripherial blood CD3+ cell, and a human peripheral blood mononuclear cell. 
     
     
         127 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 124 , wherein said monoclonal antibody, or an antigen binding fragment thereof, further comprises one or more characteristics selected from the group consisting of:
 causes an increase in cell surface calreticulin expression on human tumor cells;   causes an increase in adenosine triphosphate (ATP) release by human tumor cells;   causes an increase in high mobility group box 1 (HMGB1) release by human tumor cells;   causes an increase in annexin A1 release by human tumor cells;   causes an increase in Type I Interferon release by human tumor cells;   causes an increase in C-X-C Motif Chemokine Ligand 10 (CXCL10) release by human tumor cells;   causes an increase in cell surface protein disulfide-isomerase A3 (PDIA3) expression on human tumor cells;   causes an increase in cell surface heat shock protein 70 (HSP70) expression on human tumor cells; and   causes an increase in cell surface heat shock protein 90 (HSP90) expression on human tumor cells.   
     
     
         128 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 124 , wherein the antibody, or antigen binding fragment thereof, is a chimeric or humanized antibody; or wherein the monoclonal antibody, or antigen binding fragment thereof cross-reacts with one or more species homologs of CD47. 
     
     
         129 . A method of treating a disease comprising an autoimmune disease, an autoinflammatory disease, an inflammatory disease, a cardiovascular disease, or a cancer in a subject, comprising administration of the monoclonal antibody or antigen-binding fragment thereof, of  claim 124 , wherein the subject comprises a human or companion/pet animal, a working animal, a sport animal, a zoo animal, or a valuable animal kept in captivity. 
     
     
         130 . The method of  claim 129 , wherein the disease 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, urothelial 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, a sarcoma, ischemia-reperfusion injury, heart failure, arthritis, rheumatoid arthritis, multiple sclerosis, psoriasis, psoriatic arthritis, Crohn's disease, inflammatory bowel disease, ulcerative colitis, lupus, systemic lupus erythematous, juvenile rheumatoid arthritis, juvenile idiopathic arthritis, Grave's disease, Hashimoto's thyroiditis, Addison's disease, celiac disease, dermatomyositis, multiple sclerosis, myasthenia gravis, pernicious anemia, Sjogren syndrome, type I diabetes, vasculitis, uveitis, atherosclerosis, and ankylosing spondylitis. 
     
     
         131 . The monoclonal antibody, or antigen binding fragment thereof, of  claim 124 , wherein the monoclonal antibody, or antigen binding fragment thereof, further exhibits pH-dependent binding to CD47 present on a cell.