Novel anti-cd38 antibodies for the treatment of cancer

Abstract

Antibodies, humanized antibodies, resurfaced antibodies, antibody fragments, derivatized antibodies, and conjugates of same with cytotoxic agents, which specifically bind to CD38, are capable of killing CD38 30 cells by apoptosis, antibody-dependent cell-mediated cytotoxicity (ADCC), and/or complement-dependent cytotoxicity (CDC). Said antibodies and fragments thereof may be used in the treatment of tumors that express CD38 protein, such as multiple myeloma, chronic lymphocytic leukemia, chronic rnyelogenous leukemia, acute myelogenous leukemia, or acute lymphocytic leukemia, or the treatment of autoimmune and inflammatory diseases such as systemic lupus, rheumatoid arthritis, multiple sclerosis, erythematosus, and asthma. Said derivatized antibodies may be used in the diagnosis and imaging of tumors that express elevated levels of CD38, Also provided are cytotoxic conjugates comprising a cell binding agent and a cytotoxic agent, therapeutic compositions comprising the conjugate, methods for using the conjugates in the inhibition of cell growth and the treatment of disease, and a kit comprising the cytotoxic conjugate. In particular, the cell binding agent is a monoclonal antibody, and epitope-binding fragments thereof, that recognizes and binds the CD38 protein.

1 . An antibody or epitope-binding fragment thereof that specifically binds CD38, characterized in that said antibody or epitope-binding fragment thereof comprises at least one heavy chain and at least one light chain, wherein
 a) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 1, 2, and 3, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 4, 5, and 6, or wherein   b) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 7, 8, and 9, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 10, 11, and 12, or wherein   c) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 13, 14, and 15, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 16, 17, and 18, or wherein   d) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 19, 20, and 21, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 22, 23, and 24, or wherein   e) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 25, 26, and 27, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 28, 29, and 30, or wherein   f) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 31, 32, and 33, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 34, 35, and 36, or wherein   g) said heavy chain comprises a heavy chain variable region having an amino acid sequence of SEQ ID NO: 66 and said light chain comprises a light chain variable region having an amino asic sequence of SEQ ID NO 62 or SEQ ID NO: 64.   
     
     
         2 - 13 . (canceled) 
     
     
         14 . An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof binds CD38 with a k D  of 3×10 −9  M or lower. 
     
     
         15 - 16 . (canceled) 
     
     
         17 . An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof comprises a light chain variable region having an amino acid sequence selected from the group consisting of SEQ ID NOS: 38, 40, 42, 44, 46, and 48. 
     
     
         18 . An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence selected from the group consisting of SEQ ID NOS: 50, 52, 54, 56, 58, 60. 
     
     
         19 . (canceled) 
     
     
         20 . An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof comprises a light chain variable region having an amino acid sequence consisting of SEQ ID NO: 38 and a heavy chain variable region having an amino acid sequence of SEQ ID NO: 50. 
     
     
         21 - 22 . (canceled) 
     
     
         23 . An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof comprises a light chain variable region having an amino acid sequence of SEQ ID NO: 40 and a heavy chain variable region having an amino acid sequence of SEQ ID NO: 52. 
     
     
         24 - 25 . (canceled) 
     
     
         26 . An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof comprises a light chain variable region having an amino acid sequence of SEQ ID NO: 42 and a heavy chain variable region having an amino acid sequence of 54. 
     
     
         27 - 28 . (canceled) 
     
     
         29 . An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof comprises a light chain variable region having an amino acid sequence of SEQ ID NO: 44 and a heavy chain variable region of SEQ ID NO: 56. 
     
     
         30 - 31 . (canceled) 
     
     
         32 . An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof comprises a light chain variable region having an amino acid sequence of SEQ ID NO: 46 and a heavy chain variable region having an amino acid sequence of SEQ ID NO: 58. 
     
     
         33 - 34 . (canceled) 
     
     
         35 . An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof comprises a light chain variable region having an amino acid sequence of SEQ ID NO: 48 and a heavy chain variable region having and amino acid sequences of SEQ ID NO: 60. 
     
     
         36 . (canceled) 
     
     
         37 . An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof is produced by a hybridoma cell line selected from the group of hybridoma cell lines deposited at the American Type Culture Collection (10801 University Bld, Manassas, Va., 20110-2209, USA), on Jun. 21, 2006, under the deposit numbers PTA-7667, PTA-7669, PTA-7670, PTA-7666, PTA-7668, and PTA-7671. 
     
     
         38 . An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof comprises at least one human constant region. 
     
     
         39 . An antibody or epitope-binding fragment thereof according to  claim 38 , characterized in that said constant region is the human IgG1/IgKappa constant region. 
     
     
         40 . An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof is a humanized or resurfaced antibody. 
     
     
         41 - 47 . (canceled) 
     
     
         48 . An A humanized or resurfaced antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence represented by SEQ ID NO: 72 and a light chain variable region having an amino acid sequence selected from the group consisting of SEQ ID NOS: 68 and 70. 
     
     
         49 - 50 . (canceled) 
     
     
         51 . An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof is a Fab, Fab′, F(ab′)2 or Fv fragment. 
     
     
         52 - 64 . (canceled) 
     
     
         65 . A method of treating cancer or autoimmune disease comprising administering to a patient an antibody or epitope-binding fragment thereof that specifically binds CD38, characterized in that said antibody or epitope-binding fragment thereof comprises at least one heavy chain and at least one light chain, wherein
 a) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 1, 2, and 3, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 4, 5, and 6, or wherein   b) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 7, 8, and 9, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 10, 11, and 12, or wherein   c) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 13, 14, and 15, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 16, 17, and 18, or wherein   d) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 19, 20, and 21, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 22, 23, and 24, or wherein   e) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 25, 26, and 27, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 28, 29, and 30, or wherein   f) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 31, 32, and 33, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 34, 35, and 36, or wherein   g) said heavy chain comprises a heavy chain variable region having an amino acid sequence of SEQ ID NO: 66 and said light chain comprises a light chain variable region having an amino asic sequence of SEQ ID NO 62 or SEQ ID NO: 64.   
     
     
         66 - 71 . (canceled) 
     
     
         72 . A method of diagnosing a cancer in a subject known to or suspected to have a cancer, said method comprising:
 a) contacting cells of said patient with an antibody or epitope-binding fragment thereof that specifically binds CD38, characterized in that said antibody or epitope-binding fragment thereof comprises at least one heavy chain and at least one light chain, wherein
 i) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 1, 2, and 3, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 4, 5, and 6, or wherein 
 ii) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 7, 8, and 9, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 10, 11, and 12, or wherein 
 iii) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 13, 14, and 15, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 16, 17, and 18, or wherein 
 iv) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 19, 20, and 21, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 22, 23, and 24, or wherein 
 v) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 25, 26, and 27, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 28, 29, and 30, or wherein 
 vi) said heavy chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 31, 32, and 33, and wherein said light chain comprises three sequential complementarity-determining regions having amino acid sequences represented by SEQ ID NOS: 34, 35, and 36, or wherein 
 vii) said heavy chain comprises a heavy chain variable region having an amino acid sequence of SEQ ID NO: 66 and said light chain comprises a light chain variable region having an amino asic sequence of SEQ ID NO 62 or SEQ ID NO: 64, 
   b) measuring the binding of said antibody or epitope-binding fragment thereof to said cells, and   c) comparing the expression in part (b) with that of a normal reference subject or standard.   
     
     
         73 - 75 . (canceled) 
     
     
         76 . A polynucleotide encoding a polypeptide selected from the group consisting of SEQ ID NOS: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, and 72. 
     
     
         77 - 79 . (canceled)