Antagonistic CD40 Monoclonal Antibodies and Uses Thereof

Abstract

The disclosure provides for antibodies that bind CD40, including a humanized antibody and a chimeric antibody with different Fc domains. The antibodies bind CD40 and do not exhibit CD40 agonist activity. The antibodies may comprise a modified IgG1 Fc domain, and exhibit minimal activation of immature dendritic cells. Compositions comprising antibodies, methods of use for treatment of diseases involving CD40 activity, and use in the preparation of a medicament for treatment of a disease involving CD40 activity are provided.

We claim: 
     
         1 . An isolated antibody, or antigen binding portion thereof, that specifically binds to human CD40, wherein said antibody comprises a heavy chain, and a light chain, wherein:
 said heavy chain comprises a CDR1 comprising GYTFTDLSMHW (SEQ ID NO: 1), a CDR2 comprising YITPSSGYTAYNQKFKG (SEQ ID NO: 2), a CDR3 comprising LILQRGAY (SEQ ID NO: 3) and a human heavy chain constant region; and   said light chain comprises a CDR1 comprising RASKNVDSYGNSFMHW (SEQ ID NO: 4), a CDR2 comprising RASNLES (SEQ ID NO: 5), and a CDR3 comprising QQSNEDPLT (SEQ ID NO: 6) and a human light chain constant region, and   said human heavy chain constant region is a human IgG1 Fc domain comprising either (1) a mutation at Kabat position 238 that reduces binding to Fc-gamma-receptors (FcgRs), wherein proline 238 (P238) is mutated to one of the residues selected from the group consisting of lysine, serine, alanine, arginine and tryptophan, and wherein the antibody or antigen binding portion has reduced FcgR binding; or (2) or an alanine substituted at Kabat position 297.   
     
     
         2 . The isolated antibody or antigen binding portion thereof of  claim 1 , wherein the antibody or antigen binding portion thereof antagonizes activities of CD40. 
     
     
         3 . The isolated antibody or antigen binding portion thereof of  claim 1 , wherein said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 7 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 8. 
     
     
         4 . The isolated antibody or antigen binding portion thereof of  claim 1 , comprising a human IgG1 Fc domain comprising a mutation at Kabat position 238 that reduces binding to Fc-gamma-receptors (FcgRs), wherein proline 238 (P238) is mutated to one of the residues selected from the group consisting of lysine, serine, alanine, arginine and tryptophan, and wherein the antibody or antigen binding portion has reduced binding to at least one low affinity FcgR. 
     
     
         5 . The isolated antibody or antigen binding portion thereof of  claim 4 , wherein P238 is mutated to lysine. 
     
     
         6 . The isolated antibody or antigen binding portion thereof of  claim 4 , wherein the Fe domain comprises an amino acid sequence selected from: SEQ ID NO: 98 (IgG1a-P238K (-C-term Lys)), SEQ ID NO: 9 (IgG1a-P238K), SEQ ID NO: 99 (CH1-IgG1a-P238K (-C-term Lys)), SEQ ID NO: 10 (CH1-IgG1a-P238K), SEQ ID NO: 100 (IgG1f-P238K (-C-term Lys)), SEQ ID NO: 11 (IgG1f-P238K), SEQ ID NO: 101 (CH1-IgG1f-P238K (-C-term Lys)), or SEQ ID No: 12 (CH1-IgG1f-P238K). 
     
     
         7 . The isolated antibody or antigen binding portion thereof of  claim 3 , wherein the human IgG1 Fc domain comprises the amino acid sequence of SEQ ID NO: 98 or SEQ ID NO: 100. 
     
     
         8 . The isolated antibody or antigen binding portion thereof of  claim 1 , wherein the heavy chain comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 85, SEQ ID NO: 106, SEQ ID NO: 107, and SEQ ID NO: 108; and the light chain comprises or consists of the amino acid sequence of SEQ ID NO: 88. 
     
     
         9 . The isolated antibody or antigen binding portion thereof of  claim 1 , comprising a human IgG1 Fc domain comprising an alanine substituted at Kabat position 297. 
     
     
         10 . The isolated antibody or antigen binding portion thereof of  claim 9 , wherein the Fc domain comprises an amino acid sequence selected from: SEQ ID NO: 102 (IgG1a-N297A (-C-term Lys)), SEQ ID NO: 13 (IgG1a-N297A), SEQ ID NO: 103 (CH1-IgG1a-N297A (-C-term Lys)), SEQ ID NO: 14 (CH1-IgG1a-N297A), SEQ ID NO: 104 (IgG1f-N297A (-C-term Lys)), SEQ ID NO: 15 (IgG1f-N297A), SEQ ID NO: 105 (CH1-IgG1f-N297A (-C-term Lys)), or SEQ ID NO: 16 (CH1-IgG1f-N297A). 
     
     
         11 . The isolated antibody or antigen binding portion thereof of  claim 1 , wherein the isolated antibody or antigen binding portion thereof is humanized. 
     
     
         12 . The antibody or antigen binding portion thereof of  claim 1 , wherein the antibody or antigen-binding portion thereof is linked to a therapeutic agent. 
     
     
         13 . The antibody or antigen binding portion thereof of  claim 1 , wherein the antibody or antigen-binding portion thereof is linked to a second functional moiety having a different binding specificity than said antibody or antigen binding portion thereof. 
     
     
         14 . A nucleic acid encoding an isolated antibody or antigen binding portion thereof of  claim 1 . 
     
     
         15 . An expression vector comprising the nucleic acid molecule of  claim 14 . 
     
     
         16 . A cell transformed with the expression vector of  claim 15 . 
     
     
         17 . A method of preparing an anti-human CD40 antibody, or antigen binding portion thereof, comprising:
 a) expressing the antibody, or antigen binding portion thereof, in the cell of  claim 16 ; and   b) isolating the antibody, or antigen binding portion thereof, from the cell.   
     
     
         18 . A pharmaceutical composition comprising: a) the antibody, or antigen binding portion thereof, of  claim 1 ; and b) a pharmaceutically acceptable carrier. 
     
     
         19 . A method of treating or preventing an immune response in a subject comprising administering to the subject the antibody, or the antigen binding portion thereof, of  claim 1 . 
     
     
         20 . The method of  claim 19 , wherein the subject has a disease selected from the group consisting of: Addison's disease, allergies, anaphylaxis, ankylosing spondylitis, asthma, atherosclerosis, atopic allergy, autoimmune diseases of the ear, autoimmune diseases of the eye, autoimmune hepatitis, autoimmune parotitis, bronchial asthma, coronary heart disease, Crohn's disease, diabetes, epididymitis, glomerulonephritis, Graves' disease, Guillain-Barre syndrome, Hashimoto's disease, hemolytic anemia, idiopathic thrombocytopenic purpura, inflammatory bowel disease, immune response to recombinant drug products (e.g., Factor VII in hemophiliacs), systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, pemphigus, psoriasis, rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's syndrome, spondyloarthropathies, thyroiditis, transplant rejection, vasculitis, and ulcerative colitis.