Humanized Antibodies to Cluster of Differentiation 3 (CD3)

Abstract

Humanized monoclonal antibodies (mAbs) or fragments thereof, to human Cluster of Differentiation 3 (CD3), which confer improved immune stimulation and stability, are disclosed. Pharmaceutical compositions comprising said mAbs and methods of treatment and optionally also prevention of diseases and disorders, such as autoimmune disorders, infectious diseases, and transplant rejection, that are susceptible to amelioration by binding to CD3, are also disclosed.

1 . A monoclonal antibody (mAb), or fragment thereof, that binds specifically to Cluster of Differentiation 3 (CD3), wherein said antibody comprises:
 i. a humanized framework comprising the mutations (using Kabat numbering system) Q5V, A9S, A12K, R13K, K38R, R40A, Q43K, K66R, S75T, T83R, S87T, T108L and L109V relative to the sequence of the heavy (H) chain variable (V) region set forth in SEQ ID NO: 7 and the mutations I10T, M11L, A13L, K18R, V19A, T22S, S40P, T42K, S43A, A60S, H61R, S70D, S72T, G77S, M78L, E79Q, A80P, A83F, S100G, L104V and N107K relative to the sequence of the light kappa (κ) chain V region set forth in SEQ ID NO: 8; and   ii. the six CDRs set forth in SEQ ID NOs: 1-6.   
     
     
         2 . The mAb according to  claim 1  comprising a H chain constant domain selected from the group consisting of IgG1, IgG2, IgG3, IgG4, IgA, and IgM. 
     
     
         3 . The mAb according to  claim 1  comprising a human IgG4 constant region comprising a Ser241Pro mutation. 
     
     
         4 . The mAb or fragment according to any one of  claims 1 - 3  comprising a variable heavy (VH) chain set forth in SEQ ID NO: 9. 
     
     
         5 . The mAb or fragment according to any one of  claims 1 - 3  comprising a variable kappa (Vκ) chain set forth in SEQ ID NO: 10. 
     
     
         6 . The mAb or fragment according to any one of  claims 1 - 3  comprising a VH chain set forth in SEQ ID NO: 9 and a Vκ chain set forth in SEQ ID NO: 10. 
     
     
         7 . The mAb or fragment according to any one of  claims 1 - 3  comprising a full-length H chain according to SEQ ID NO: 11 and a full-length K chain according to SEQ ID NO: 12. 
     
     
         8 . The mAb or fragment according to any one of  claims 1 - 3  comprising a H chain sequence having at least 95% identity to SEQ ID NO: 9 and a K chain sequence having at least 95% identity to SEQ ID NO: 10. 
     
     
         9 . The mAb or fragment according to any one of  claims 1 - 8  comprising a VH chain encoded by a polynucleotide sequence comprising a sequence set forth in SEQ ID NO: 13, or such polynucleotide sequence having at least 70%, 75%, 80%, 85%, 90% or 95% identity while still encoding said polypeptide. 
     
     
         10 . The mAb or fragment according to any one of  claims 1 - 8  comprising a Vκ chain encoded by a polynucleotide sequence comprising a sequence set forth in SEQ ID NO: 14, or such polynucleotide sequence having at least 70%, 75%, 80%, 85%, 90% or 95% identity while still encoding said polypeptide. 
     
     
         11 . The mAb or fragment according to any one of  claims 1 - 10 , comprising at least the V region. 
     
     
         12 . The mAb or fragment according to any one of  claims 1 - 11 , wherein the mAb or fragment is a mAb fragment. 
     
     
         13 . The mAb fragment according to  claim 12  comprising a VH chain set forth in SEQ ID NO: 9 or a Vκ chain set forth in SEQ ID NO: 10. 
     
     
         14 . The mAb fragment according to  claim 12  comprising a VH chain set forth in SEQ ID NO: 9 and a Vκ chain set forth in SEQ ID NO: 10. 
     
     
         15 . The mAb fragment according to  claim 12 , comprising at least the V regions, wherein the mAb fragment is selected from the group consisting of: Fab, Fab′, F(ab′) 2 , scFv, and dsFv antibody. 
     
     
         16 . The mAb or fragment according to any of the above claims which is a bispecific or multi-specific antibody. 
     
     
         17 . A pharmaceutical composition comprising at least one mAb or fragment according to any one of  claims 1 - 16 . 
     
     
         18 . The pharmaceutical composition according to  claim 17  further comprising an excipient, diluent or carrier. 
     
     
         19 . The pharmaceutical composition according to  claim 17  formulated for oral or mucosal administration. 
     
     
         20 . The pharmaceutical composition according to  claim 17  for treating a disease or disorder susceptible to treatment or amelioration through binding to CD3. 
     
     
         21 . A method of treating a disease or disorder susceptible to treatment or amelioration through binding to CD3 comprising administering to a subject in need thereof a pharmaceutical composition according to  claim 17 . 
     
     
         22 . The method according to  claim 21 , wherein the disease is an autoimmune disease selected from the group consisting of Alopecia Areata, Lupus, Ankylosing Spondylitis, Meniere's Disease, Antiphospholipid Syndrome, Mixed Connective Tissue Disease, Autoimmune Addison's Disease, Multiple Sclerosis, Autoimmune Hemolytic Anemia, Myasthenia Gravis, Autoimmune Hepatitis, Pemphigus Vulgaris, Behcet's Disease, Pernicious Anemia, Bullous Pemphigoid, Polyarthritis Nodosa, Cardiomyopathy, Polychondritis, Celiac Sprue-Dermatitis, Polyglandular Syndromes, Chronic Fatigue Syndrome (CFIDS), Polymyalgia Rheumatica, Chronic Inflammatory Demyelinating, Polymyositis and Dermatomyositis, Chronic Inflammatory Polyneuropathy, Primary Agammaglobulinemia, Churg-Strauss Syndrome, Primary Biliary Cirrhosis, Cicatricial Pemphigoid, Psoriasis, CREST Syndrome, Raynaud's Phenomenon, Cold Agglutinin Disease, Reiter's Syndrome, Crohn's Disease, Rheumatic Fever, Discoid Lupus, Rheumatoid Arthritis, Essential Mixed, Cryoglobulinemia Sarcoidosis, Fibromyalgia, Scleroderma, Grave's Disease, Sjogren's Syndrome, Guillain-Barre, Stiff-Man Syndrome, Hashimoto's Thyroiditis, Takayasu Arteritis, Idiopathic Pulmonary Fibrosis, Temporal Arteritis/Giant Cell Arteritis, Idiopathic Thrombocytopenia Purpura (ITP), Ulcerative Colitis, IgA Nephropathy, Uveitis, Insulin Dependent Diabetes (Type 1), non-Insulin-Dependent Diabetes (Type II), Vasculitis, Lichen Planus, Vitiligo, and Hepatitis. 
     
     
         23 . The method according to  claim 21 , wherein the disorder is associated with an immune response associated with cell, tissue or organ transplantation. 
     
     
         24 . The method according to  claim 21 , wherein the disease is hepatitis 
     
     
         25 . The method according to  claim 24 , wherein the cause of hepatitis is selected from the group consisting of: an infectious agent, a toxin, a substance which is toxic to the liver with excessive intake; non-alcoholic steatohepatitis (NASH); a liver disease associated with inflammatory bowel disease, hyperlipidemia. 
     
     
         26 . The method according to  claim 25 , wherein the infectious agent is a virus selected from the group consisting of: hepatitis A, B, C, D or E; a herpes virus, a yellow fever virus, a human immunodeficiency virus (HIV), and an adenovirus.