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+ 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 myelogenous 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 is capable of killing a CD38 +  cell by apoptosis, antibody-dependent cell-mediated cytotoxicity (ADCC), and complement-dependent cytotoxicity (CDC). 
     
     
         2 : An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof is capable of killing said CD38 +  cell by apoptosis in the absence of stroma cells or stroma-derived cytokines. 
     
     
         3 : An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said antibody or epitope-binding fragment thereof is a monoclonal antibody. 
     
     
         4 : An antibody or epitope-binding fragment thereof according to  claim 1 , characterized in that said CD38 +  cell is a lymphoma cell, a leukemia cell, or a multiple myeloma cell. 
     
     
         5 : The antibody or epitope-binding fragment thereof of  claim 4 , characterized in that said CD38 +  cell is a non-Hodgkin's lymphoma (NHL) cell, a Burkitt's lymphoma (BL) cell, a multiple myeloma (MM) cell, a B chronic lymphocytic leukemia (B-CLL) cell, a B and T acute lymphocytic leukemia (ALL) cell, a T cell lymphoma (TCL) cell, an acute myeloid leukemia (AML) cell, a hairy cell leukemia (HCL) cell, a Hodgkin's Lymphoma (HL) cell, or a chronic myeloid leukemia (CML) cell. 
     
     
         6 - 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 - 37 . (canceled) 
     
     
         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 : A humanized or resurfaced antibody or epitope-binding fragment thereof according to  claim 40 , characterized in that said humanized or resurfaced antibody or epitope-binding fragment thereof comprises at least one heavy chain and at least one light chain, wherein said heavy chain comprises three sequential complementarity-determining regions of SEQ ID NOS: 1, 2, and 3, and wherein said light chain comprises three sequential complementarity-determining regions of SEQ ID NOS: 4, 5, and 6. 
     
     
         42 : A humanized or resurfaced antibody or epitope-binding fragment thereof according to  claim 40 , characterized in that said humanized or resurfaced antibody or epitope-binding fragment thereof comprises at least one heavy chain and at least one light chain, wherein said heavy chain comprises three sequential complementarity-determining regions of SEQ ID NOS: 7, 8, and 9, and wherein said light chain comprises three sequential complementarity-determining regions of SEQ ID NOS: 10, 11, and 12. 
     
     
         43 - 45 . (canceled) 
     
     
         46 : A humanized or resurfaced antibody or epitope-binding fragment thereof according to  claim 40 , characterized in that said humanized or resurfaced antibody or epitope-binding fragment thereof comprises at least one heavy chain and at least one light chain, wherein said heavy chain comprises three sequential complementarity-determining regions of SEQ ID NOS: 19, 20, and 21, and wherein said light chain comprises three sequential complementarity-determining regions SEQ ID NOS: 22, 23, and 24. 
     
     
         47 : A humanized or resurfaced antibody or epitope-binding fragment thereof according to  claim 40 , characterized in that said humanized or resurfaced antibody or epitope-binding fragment thereof comprises at least one heavy chain and at least one light chain, wherein said heavy chain comprises three sequential complementarity-determining regions of SEQ ID NOS: 25, 26, and 27, and wherein said light chain comprises three sequential complementarity-determining regions of SEQ ID NOS: 28, 29, and 30. 
     
     
         48 : A humanized or resurfaced antibody or epitope-binding fragment thereof according to  claim 47 , characterized in that said humanized or resurfaced antibody or epitope-binding fragment thereof comprises a heavy chain variable region comprising SEQ ID NO: 72 and a light chain variable region comprising SEQ ID NO: 68 or SEQ ID NO: 70. 
     
     
         49 . (canceled) 
     
     
         50 : A humanized or resurfaced antibody or epitope-binding fragment thereof according to  claim 40 , characterized in that said humanized or resurfaced antibody or epitope-binding fragment thereof comprises at least one heavy chain and at least one light chain, wherein said heavy chain comprises three sequential complementarity-determining regions of SEQ ID NOS: 31, 32, and 33, and wherein said light chain comprises three sequential complementarity-determining regions of SEQ ID NOS: 34, 35, and 36. 
     
     
         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 : A conjugate comprising the antibody or epitope-binding fragment thereof according to  claim 1  linked to a cytotoxic agent. 
     
     
         53 : The conjugate of  claim 52 , characterized in that said cytotoxic agent is selected from the group consisting of a maytansinoid, a small drug, a tomaymycin derivative, a leptomycin derivative, a prodrug, a taxoid, CC-1065 and a CC-1065 analog. 
     
     
         54 : The conjugate of  claim 53 , characterized in that said cytotoxic agent is the maytansine DM1 of formula: 
       
         
           
           
               
               
           
         
       
     
     
         55 : The conjugate of  claim 53 , characterized in that said cytotoxic agent is the maytansine DM4 of formula: 
       
         
           
           
               
               
           
         
       
     
     
         56 : The conjugate of  claim 53 , characterized in that said cytotoxic agent is a tomaymycin derivative selected from the group consisting of:
 8,8′-[1,3-benzenediylbis(methyleneoxy)]-bis[(S)-2-eth-(E)-ylidene-7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[5-methoxy-1,3-benzenediylbis(methyleneoxy)]-bis[(S)-2-eth-(E)-ylidene-7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[1,5-pentanediylbis(oxy)]-bis[(S)-2-eth-(E)-ylidene-7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[1,4-butanediylbis(oxy)]-bis[(S)-2-eth-(E)-ylidene-7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[3-methyl-1,5-pentanediylbis(oxy)]-bis[(S)-2-eth-(E)-ylidene-7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[2,6-pyridinediylbis(oxy)]-bis[(S)-2-eth-(E)-ylidene-7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[4-(3-tert-butoxycarbonylaminopropyloxy)-2,6-pyridinediylbis-(methyleneoxy)]-bis[(S)-2-eth-(E)-ylidene-7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[5-(3-aminopropyloxy)-1,3-benzenediylbis(methyleneoxy)]-bis[(S)-2-eth-(E)-ylidene-7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[5-(N-methyl-3-tert-butoxycarbonylaminopropyl)-1,3-benzenediylbis-(methyleneoxy)]-bis[(S)-2-eth-(E)-ylidene-7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-{5-[3-(4-methyl-4-methyldisulfanyl-pentanoylamino)propyloxy]-1,3-benzenediylbis(methyleneoxy)}-bis[(S)-2-eth-(E)-ylidene-7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[5-acetylthiomethyl-1,3-benzenediylbis(methyleneoxy)]-bis[(S)-2-methylene-7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   bis-{2-[(S)-2-methylene-7-methoxy-5-oxo-1,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-8-yloxy]-ethyl}-carbamic acid tert-butyl ester   8,8′-[3-(2-acetylthioethyl)-1,5-pentanediylbis(oxy)]-bis[(S)-2-methylene-7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[5-(N-4-mercapto-4,4-dimethylbutanoyl)amino-1,3-benzenediylbis(methyleneoxy)]-bis[7-methoxy-2-methylene-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[5-(N-4-methyldithio-4,4-dimethylbutanoyl)-amino-1,3-benzenediylbis(methyleneoxy)]-bis[7-methoxy-2-methylene-1,2,3, 11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[5-(N-methyl-N-(2-mercapto-2,2-dimethylethyl)amino-1,3-benzenediyl(methyleneoxy)]-bis[7-methoxy-2-methyl ene-1,2,3, 11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[5-(N-methyl-N-(2-methyldithio-2,2-dimethylethyl)amino-1,3-benzenediyl(methyleneoxy)]-bis[7-methoxy-2-methylene-1,2,3, 11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[(4-(2-(4-mercapto-4-methyl)-pentanamido-ethoxy)-pyridin-2,6-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3, 11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[(1-(2-(4-methyl-4-methyldisulfanyl)-pentanamido-ethoxy)-benzene-3, 5-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3, 11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[(4-(3-(4-methyl-4-methyldisulfanyl)-pentanamido-propoxy)-pyridin-2, 6-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3, 11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[(4-(4-(4-methyl-4-methyldisulfanyl)-pentanamido-butoxy)-pyridin-2,6-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3,11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[(4-(3-[4-(4-methyl-4-methyldisulfanyl-pentanoyl)-piperazin-1-yl]-propyl)-pyridin-2,6-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3, 11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[(1-(3-[4-(4-methyl-4-methyldisulfanyl-pentanoyl)-piperazin-1-yl]-propyl)-benzene-3, 5-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3, 11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[(4-(2-{2-[2-(4-methyl-4-methyldisulfanyl-pentanoylamino)-ethoxy]-ethoxy}-ethoxy)-pyridin-2,6-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3,11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[(1-(2-{2-[2-(2-{2-[2-(4-methyl-4-methyldisulfanyl-pentanoylamino)-ethoxy]-ethoxy}-ethoxy)-ethoxy]-ethoxy}-ethoxy)-benzene-3, 5-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3, 11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[(1-(2-{2-[2-(4-methyl-4-methyldisulfanyl-pentanoylamino)-ethoxy]-ethoxy}-ethoxy)-benzene-3,5-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3, 11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[(4-(2-{2-[2-(2-{2-[2-(4-methyl-4-methyldisulfanyl-pentanoylamino)-ethoxy]-ethoxy}-ethoxy)-ethoxy]-ethoxy}-ethoxy)-pyridin-2,6-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3, 11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[(1-(2-[methyl-(2-methyl-2-methyldisulfanyl-propyl)-amino]-ethoxy)-benzene-3,5-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3,11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[(4-(3-[methyl-(4-methyl-4-methyldisulfanyl-pentanoyl)-amino]-propyl)-pyridin-2,6-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3,11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   8,8′-[(4-(3-[methyl-(2-methyl-2-methyldisulfanyl-propyl)-amino]-propyl)-pyridin-2,6-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3,11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one]   and   8,8′-[(1-(4-methyl-4-methyldisulfanyl)-pentanamido)-benzene-3,5-dimethyl)-dioxy]-bis[(S)-2-eth-(E)-ylidene-7-dimethoxy-1,2,3,11a-tetrahydro-pyrrolo[2,1-c][1,4]benzodiazepin-5-one].   
     
     
         57 . (canceled) 
     
     
         58 : The conjugate of  claim 53 , characterized in that the cytotoxic agent is a leptomycin derivative selected from the group consisting of:
 (2-Methyl sulfanyl-ethyl)-amid of (2E,10E,12E,16Z,18E)-(R)-6-Hydroxy-3,5, 7,9,11,15,17-heptamethyl-19-((2S,3S)-3-methyl-6-oxo-3,6-dihydro-2H-pyran-2-yl)-8-oxo-nonadeca-2,10,12,16,18-pentaenoic acid (2-methylsulfanyl-ethyl)-amid   Bis-[(2-mercaptoethyl)-amid of (2E,10E,12E,16Z,18E)-(R)-6-hydroxy-3,5,7,9, 11,15,17-heptamethyl-19-((2S,3 S)-3-methyl-6-oxo-3,6-dihydro-2H-pyran-2-yl)-8-oxo-nonadeca-2,10,12,16,18-pentaenoic acid]   (2-Mercapto-ethyl)-amid of (2E,10E,12E,16Z,18E)-(R)-6-hydroxy-3,5,7,9,11, 15,17-heptamethyl-19-((2S,3 S)-3-methyl-6-oxo-3,6-dihydro-2H-pyran-2-yl)-8-oxo-nonadeca-2,10,12,16,18-pentaenoic acid   (2-Methyldisulfanyl-ethyl)-amid of (2E,10E,12E,16Z,18E)-(R)-6-hydroxy-3,5,7,9,11,15,17-heptamethyl-19-((2S,3 S)-3-methyl-6-oxo-3,6-dihydro-2H-pyran-2-yl)-8-oxo-nonadeca-2,10,12,16,18-pentaenoic acid   (2-Methyl-2-methyldisulfanyl-propyl)-amid of (2E,10E,12E,16Z,18E)-(R)-6-hydroxy-3,5,7,9,11,15,17-heptamethyl-19-((2S,3 S)-3-methyl-6-oxo-3,6-dihydro-2H-pyran-2-yl)-8-oxo-nonadeca-2,10,12,16,18-pentaenoic acid   and   (2-Mercapto-2-methyl-propyl)-amid of (2E,10E,12E,16Z,18E)-(R)-6-hydroxy-3,5,7,9,11,15,17-heptamethyl-19-((2S,3 S)-3-methyl-6-oxo-3,6-dihydro-2H-pyran-2-yl)-8-oxo-nonadeca-2,10,12,16,18-pentaenoic acid.   
     
     
         59 - 60 . (canceled) 
     
     
         61 : A pharmaceutical composition comprising (a) the antibody or epitope-binding fragment thereof according to  claim 1  or the conjugate according to  claim 52 , and (b) a pharmaceutically acceptable carrier or excipients. 
     
     
         62 : The pharmaceutical composition of claim  59 , comprising a further therapeutic agent. 
     
     
         63 : The pharmaceutical composition of claim  60 , characterized in that the further therapeutic agent is an antagonist of epidermal-growth factor (EGF), fibroblast-growth factor (FGF), hepatocyte growth factor (HGF), tissue factor (TF), protein C, protein S, platelet-derived growth factor (PDGF), heregulin, macrophage-stimulating protein (MSP) or vascular endothelial growth factor (VEGF), a receptor for epidermal-growth factor (EGF), fibroblast-growth factor (FGF), hepatocyte growth factor (HGF), tissue factor (TF), protein C, protein S, platelet-derived growth factor (PDGF), heregulin, macrophage-stimulating protein (MSP), vascular endothelial growth factor (VEGF), HER2 receptor, HER3 receptor, c-MET, or other receptor tyrosine kinase. 
     
     
         64 : The pharmaceutical composition of claim  60 , characterized in that the further therapeutic agent is an antibody directed against a cluster of differentiation antigen selected from a group consisting of: CD3, CD14, CD19, CD20, CD22, CD25, CD28, CD30, CD33, CD36, CD40, CD44, CD52, CD55, CD59, CD56, CD70, CD79, CD80, CD103, CD134, CD137, CD138, and CD152. 
     
     
         65 : A method of treating cancer or autoimmune disease comprising administering to a patient the antibody or epitope-binding fragment thereof according to  claim 1 , or a conjugate according to  claim 52 . 
     
     
         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 according to  claim 1 ,   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 : One or more polynucleotides encoding an antibody or epitope-binding fragment thereof, wherein the antibody or epitope-binding fragment thereof comprises:
 a) a heavy chain variable region (V H ) comprising SEQ ID NO: 50 and a light chain variable region (V L ) comprising SEQ ID NO: 38;   b) a heavy chain variable region (V H ) comprising SEQ ID NO: 52 and a light chain variable region (V L ) comprising SEQ ID NO: 40;   c) a heavy chain variable region (V H ) comprising SEQ ID NO: 66 and a light chain variable region (V L ) comprising SEQ ID NO: 62 or 64;   d) a heavy chain variable region (V H ) comprising SEQ ID NO: 56 and a light chain variable region (V L ) comprising 44;   e) a heavy chain variable region (V H ) comprising SEQ ID NO: 72 and a light chain variable region (V L ) comprising SEQ ID NO: 68 or 70;   f) a heavy chain variable region (V H ) comprising SEQ ID NO: 60 and a light chain variable region (V L ) comprising 48.   
     
     
         77 . (canceled) 
     
     
         78 : A recombinant vector comprising the one or more polynucleotides of  claim 76 . 
     
     
         79 : A host cell comprising a vector of  claim 78 .