Anti-CD19 Antibodies

Abstract

The present invention provides humanized, chimeric and human anti-CD19 antibodies, anti-CD19 antibody fusion proteins, and fragments thereof that bind to a human B cell marker. Such antibodies, fusion proteins and fragments thereof are useful for the treatment and diagnosis of various B-cell disorders, including B-cell malignancies and autoimmune diseases. In more particular embodiments, the humanized anti-CD19 antibodies may comprise one or more framework region amino acid substitutions designed to improve protein stability, antibody binding and/or expression levels. In a particularly preferred embodiment, the substitutions comprise a Ser9lPhe substitution in the hA19 VH sequence.

What is claimed is: 
     
         1 . A method of treating an autoimmune disease comprising administering a bispecific antibody comprising:
 a) a first anti-CD19 antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises (i) the light chain complementarity determining region CDR sequences CDR1 of SEQ ID NO: 16 (KASQSVDYDGDSYLN); CDR2 of SEQ ID NO: 17 (DASNLVS); and CDR3 of SEQ ID NO: 18 (QQSTEDPWT); (ii) the heavy chain CDR sequences CDR1 of SEQ ID NO: 19 (SYWMN); CDR2 of SEQ ID NO: 20 (QIWPGDGDTNYNGKFKG) and CDR3 of SEQ ID NO: 21 (RETTTVGRYYYAMDY), and (iii) human antibody framework (FR) and constant region sequences with one or more framework region amino acid residues substituted from the corresponding framework region sequences of the parent murine antibody, wherein said substituted FR residues comprise the substitution of serine for phenylalanine at Kabat residue 91 of the heavy chain variable region; and   b) a second antibody or humanized antibody or antigen-binding fragment thereof, wherein the second antibody binds to an antigen selected from the group consisting of CD3, CD5, CD16, CD20, CD22, CD32b, CD33, CD37, CD38, CD40, and HLA-DR.   
     
     
         2 . The method of  claim 1 , wherein the autoimmune disease is selected from the group consisting of acute idiopathic thrombocytopenic purpura, chronic idiopathic thrombocytopenic purpura, dermatomyositis, Sydenham's chorea, myasthenia gravis, systemic lupus erythematosus, lupus nephritis, rheumatic fever, polyglandular syndromes, bullous pemphigoid, diabetes mellitus, Henoch-Schonlein purpura, post-streptococcal nephritis, erythema nodosum, Takayasu's arteritis, Addison's disease, rheumatoid arthritis, multiple sclerosis, sarcoidosis, ulcerative colitis, erythema multiforme, IgA nephropathy, polyarteritis nodosa, ankylosing spondylitis, Goodpasture's syndrome, thromboangitis ubiterans, Sjogren's syndrome, primary biliary cirrhosis, Hashimoto's thyroiditis, thyrotoxicosis, scleroderma, chronic active hepatitis, polymyositis, dermatomyositis, polychondritis, pemphigus vulgaris, Wegener's granulomatosis, membranous nephropathy, amyotrophic lateral sclerosis, tabes dorsalis, giant cell arteritis/polymyalgia, pernicious anemia, rapidly progressive glomerulonephritis, psoriasis, and fibrosing alveolitis. 
     
     
         3 . The method of  claim 1 , wherein the bispecific antibody is not conjugated to any therapeutic or diagnostic agent. 
     
     
         4 . The method of  claim 1 , wherein the second antibody or fragment thereof binds to CD3. 
     
     
         5 . The method of  claim 1 , wherein the second antibody or fragment thereof binds to CD16. 
     
     
         6 . The method of  claim 1 , wherein the second antibody or fragment thereof binds to HLA-DR. 
     
     
         7 . The method of  claim 3 , further comprising administering to said subject a drug selected from the group consisting of doxorubicin, methotrexate, paclitaxel, cyclophosphamide, etoposide, carmustine, vincristine, procarbazine, prednisone, bleomycin, leucovorin, phenyl butyrate, bryostatin-1 and CPT-11. 
     
     
         8 . The method of  claim 1 , wherein the anti-CD19 antibody or antigen-binding fragment thereof comprises the sequences of SEQ ID NO:7 (hA19VK) and SEQ ID NO:10 (hA19VH). 
     
     
         9 . The method of  claim 1 , wherein the bispecific antibody is conjugated to at least one therapeutic agent. 
     
     
         10 . The method of  claim 9  wherein said therapeutic agent is a drug selected from the group consisting of a vinca alkaloid, an anthracycline, a camptothecan, an epipodophyllotoxin, a taxane, a proteosome inhibitor, a nitrogen mustard, an alkyl sulfonate, a nitrosourea, an antimetabolite, an alkylating agent, a triazene, a folic acid analog, a COX-2 inhibitor, a pyrimidine analog, a purine analog, a platinum coordination complex, an antibiotic, a COX-2 inhibitor, an anti-mitotic agent, an anti-angiogenic agent and a pro-apoptotic agent. 
     
     
         11 . The method of  claim 10 , wherein said therapeutic agent is a drug selected from the group consisting of doxorubicin, methotrexate, paclitaxel, cyclophosphamide, etoposide, carmustine, vincristine, procarbazine, prednisone, bleomycin, leucovorin, phenyl butyrate, bryostatin-1 and CPT-11. 
     
     
         12 . The method of  claim 9 , wherein said therapeutic agent is a toxin selected from the group consisting of ricin, abrin, alpha toxin, saporin, onconase, ribonuclease (RNase), DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtheria toxin,  Pseudomonas  exotoxin and  Pseudomonas  endotoxin. 
     
     
         13 . The method of  claim 9 , wherein said therapeutic agent is selected from the group consisting of tumor necrosis factor (TNF), an interleukin (IL), granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), interferon-alpha, interferon-beta, and interferon-gamma. 
     
     
         14 . A method of treating an autoimmune disease comprising administering a bispecific antibody comprising:
 a) a first anti-CD19 antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises (i) the light chain complementarity determining region CDR sequences CDR1 of SEQ ID NO: 16 (KASQSVDYDGDSYLN); CDR2 of SEQ ID NO: 17 (DASNLVS); and CDR3 of SEQ ID NO: 18 (QQSTEDPWT); (ii) the heavy chain CDR sequences CDR1 of SEQ ID NO: 19 (SYWMN); CDR2 of SEQ ID NO: 20 (QIWPGDGDTNYNGKFKG) and CDR3 of SEQ ID NO: 21 (RETTTVGRYYYAMDY), and (iii) human antibody framework (FR) and constant region sequences with one or more framework region amino acid residues substituted from the corresponding framework region sequences of the parent murine antibody, wherein said substituted FR residues comprise the substitution of serine for phenylalanine at Kabat residue 91 of the heavy chain variable region; and   b) a second antibody or humanized antibody or antigen-binding fragment thereof, wherein the second antibody binds to CD3.   
     
     
         15 . A method of treating an autoimmune disease comprising administering a bispecific antibody comprising:
 a) a first anti-CD19 antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises (i) the light chain complementarity determining region CDR sequences CDR1 of SEQ ID NO: 16 (KASQSVDYDGDSYLN); CDR2 of SEQ ID NO: 17 (DASNLVS); and CDR3 of SEQ ID NO: 18 (QQSTEDPWT); (ii) the heavy chain CDR sequences CDR1 of SEQ ID NO: 19 (SYWMN); CDR2 of SEQ ID NO: 20 (QIWPGDGDTNYNGKFKG) and CDR3 of SEQ ID NO: 21 (RETTTVGRYYYAMDY), and (iii) human antibody framework (FR) and constant region sequences with one or more framework region amino acid residues substituted from the corresponding framework region sequences of the parent murine antibody, wherein said substituted FR residues comprise the substitution of serine for phenylalanine at Kabat residue 91 of the heavy chain variable region; and   b) a second antibody or humanized antibody or antigen-binding fragment thereof, wherein the second antibody binds to CD16.