Bispecific antibodies against cd3 and cd20

Abstract

Bispecific antibodies directed to CD3 and CD20 and uses of such bispecific antibodies, in particular use thereof in the treatment of diseases in which specific targeting and T cell-mediated killing of cells that express CD20 is desired.

1 - 71 . (canceled) 
     
     
         72 . A method of treating cancer comprising administering to a subject in need thereof a therapeutically effective amount of a bispecific antibody comprising:
 (a) a first binding arm comprising a first antigen-binding region which binds to CD3e and comprises a variable heavy chain (VH) region comprising the amino acid sequence set forth in SEQ ID NO: 6 and a variable light chain (VL) region comprising the amino acid sequence set forth in SEQ ID NO: 10, and   (b) a second binding arm comprising a second antigen-binding region which binds to human CD20 and comprises VH CDR1, VH CDR2, and VH CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 32, 33, and 34, respectively, and VL CDR1, VL CDR2, and VL CDR3 regions comprising the amino acid sequences set forth in SEQ ID NO: 35, the sequence DAS, and SEQ ID NO: 36, respectively.   
     
     
         73 . The method of  claim 72 , wherein the bispecific antibody comprises a first heavy chain and a second heavy chain and a first light chain and a second light chain, wherein said first heavy chain and said second heavy chain comprise at least a hinge region, a CH2 and CH3 region, wherein in said first heavy chain at least one of the amino acids in the positions corresponding to a position selected from the group consisting of T366, L368, K370, D399, F405, Y407, and K409 in a human IgG1 heavy chain has been substituted, and in said second heavy chain at least one of the amino acids in the positions corresponding to a position selected from the group consisting of T366, L368, K370, D399, F405, Y407, and K409 in a human IgG1 heavy chain has been substituted, wherein said first heavy chain and said second heavy chain are not substituted in the same position, and wherein the positions are numbered according to the EU Index. 
     
     
         74 . The method of  claim 73 , wherein (i) the amino acid in the position corresponding to F405 in a human IgG1 heavy chain is L in said first heavy chain, and the amino acid in the position corresponding to K409 in a human IgG1 heavy chain is R in said second heavy chain, or (ii) the amino acid in the position corresponding to K409 in a human IgG1 heavy chain is R in said first heavy chain, and the amino acid in the position corresponding to F405 in a human IgG1 heavy chain is L in said second heavy chain. 
     
     
         75 . The method of  claim 72 , wherein the bispecific antibody comprises a first heavy chain and a second heavy chain and a first light chain and a second light chain, wherein the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain of both said first heavy chain and said second heavy chain are F, E, and A, respectively, wherein the positions are numbered according to the EU Index. 
     
     
         76 . The method of  claim 72 , wherein the bispecific antibody comprises a first heavy chain and a second heavy chain and a first light chain and a second light chain, wherein the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain of both said first heavy chain and said second heavy chain are F, E, and A, respectively, wherein the position corresponding to F405 in a human IgG1 heavy chain of said first heavy chain is L, and the position corresponding to K409 in a human IgG1 heavy chain of said second heavy chain is R, and wherein the positions are numbered according to the EU Index. 
     
     
         77 . The method of  claim 72 , wherein the bispecific antibody is a full-length IgG1, kappa antibody. 
     
     
         78 . The method of  claim 72 , wherein the cancer is a mature B cell neoplasm. 
     
     
         79 . The method of  claim 78 , wherein the mature B cell neoplasm is selected from the group consisting of: B cell chronic lymphocytic leukemia, small lymphocytic lymphoma, B cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, mantle cell lymphoma, follicular lymphoma, cutaneous follicle center lymphoma, marginal zone lymphoma, hairy cell leukemia, diffuse large B cell lymphoma, Burkitt's lymphoma, plasmacytoma, plasma cell myeloma, post-transplant lymphoproliferative disorder, Waldenström's macroglobulinemia, malignant melanoma, and anaplastic large-cell lymphoma. 
     
     
         80 . The method of  claim 72 , wherein the cancer is diffuse large B-cell lymphoma. 
     
     
         81 . The method of  claim 72 , wherein the cancer is follicular lymphoma. 
     
     
         82 . The method of  claim 72 , wherein the method comprises further administering one or more therapeutic agents. 
     
     
         83 . A method of treating cancer comprising administering to a subject in need thereof a therapeutically effective amount of a bispecific antibody comprising:
 (a) a first binding arm comprising a first antigen-binding region which binds to CD3e, wherein said first antigen-binding region comprises a variable heavy chain (VH) region comprising the sequence set forth in SEQ ID NO: 6 and a variable light chain (VL) region comprising the sequence set forth in SEQ ID NO: 10, and   (b) a second binding arm comprising a second antigen-binding region which binds to human CD20 and comprises a VH region comprising the amino acid sequence set forth in SEQ ID NO: 27 and a VL region comprising the sequence set forth in SEQ ID NO: 28.   
     
     
         84 . The method of  claim 83 , wherein the bispecific antibody comprises a first heavy chain and a second heavy chain and a first light chain and a second light chain, wherein said first heavy chain and said second heavy chain comprise at least a hinge region, a CH2 and CH3 region, wherein in said first heavy chain at least one of the amino acids in the positions corresponding to a position selected from the group consisting of T366, L368, K370, D399, F405, Y407, and K409 in a human IgG1 heavy chain has been substituted, and in said second heavy chain at least one of the amino acids in the positions corresponding to a position selected from the group consisting of T366, L368, K370, D399, F405, Y407, and K409 in a human IgG1 heavy chain has been substituted, wherein said first heavy chain and said second heavy chain are not substituted in the same position, and wherein the positions are numbered according to the EU Index. 
     
     
         85 . The method of  claim 84 , wherein (i) the amino acid in the position corresponding to F405 in a human IgG1 heavy chain is L in said first heavy chain, and the amino acid in the position corresponding to K409 in a human IgG1 heavy chain is R in said second heavy chain, or (ii) the amino acid in the position corresponding to K409 in a human IgG1 heavy chain is R in said first heavy chain, and the amino acid in the position corresponding to F405 in a human IgG1 heavy chain is L in said second heavy chain. 
     
     
         86 . The method of  claim 83 , wherein the bispecific antibody comprises a first heavy chain and a second heavy chain and a first light chain and a second light chain, wherein the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain of both said first heavy chain and said second heavy chain are F, E, and A, respectively, wherein the positions are numbered according to the EU Index. 
     
     
         87 . The method of  claim 83 , wherein the bispecific antibody comprises a first heavy chain and a second heavy chain and a first light chain and a second light chain, wherein the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain of both said first heavy chain and said second heavy chain are F, E, and A, respectively, wherein the position corresponding to F405 in a human IgG1 heavy chain of said first heavy chain is L, and the position corresponding to K409 in a human IgG1 heavy chain of said second heavy chain is R, and wherein the positions are numbered according to the EU Index. 
     
     
         88 . The method of  claim 83 , wherein the bispecific antibody is a full-length IgG1, kappa antibody. 
     
     
         89 . The method of  claim 83 , wherein the cancer is a mature B cell neoplasm. 
     
     
         90 . The method of  claim 89 , wherein the mature B cell neoplasm is selected from the group consisting of: B cell chronic lymphocytic leukemia, small lymphocytic lymphoma, B cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, mantle cell lymphoma, follicular lymphoma, cutaneous follicle center lymphoma, marginal zone lymphoma, hairy cell leukemia, diffuse large B cell lymphoma, Burkitt's lymphoma, plasmacytoma, plasma cell myeloma, post-transplant lymphoproliferative disorder, Waldenström's macroglobulinemia, malignant melanoma, and anaplastic large-cell lymphoma. 
     
     
         91 . The method of  claim 83 , wherein the cancer is diffuse large B-cell lymphoma. 
     
     
         92 . The method of  claim 83 , wherein the cancer is follicular lymphoma. 
     
     
         93 . The method of  claim 83 , wherein the method comprises further administering one or more therapeutic agents. 
     
     
         94 . A nucleic acid encoding the heavy chain and/or light chain of a first and/or second binding arm of a bispecific antibody comprising (a) a first binding arm comprising a first antigen-binding region which binds to CD3e and comprises a variable heavy chain (VH) region comprising the amino acid sequence set forth in SEQ ID NO: 6 and a variable light chain (VL) region comprising the amino acid sequence set forth in SEQ ID NO: 10, and (b) a second binding arm comprising a second antigen-binding region which binds to human CD20, an expression vector which comprises the nucleic acid, or a host cell comprising the nucleic acid or expression vector. 
     
     
         95 . A method for detecting whether cross-linking between CD3- and CD20-expressing cells occurs in a sample derived from a patient upon administration of a bispecific antibody comprising (a) a first binding arm comprising a first antigen-binding region which binds to CD3e and comprises a variable heavy chain (VH) region comprising the amino acid sequence set forth in SEQ ID NO: 6 and a variable light chain (VL) region comprising the amino acid sequence set forth in SEQ ID NO: 10, and (b) a second binding arm comprising a second antigen-binding region which binds to human CD20, comprising the steps of:
 (i) contacting the sample with the bispecific antibody under conditions that allow for formation of a complex between said bispecific antibody and the CD3- and CD20-expressing cells; and   (ii) analyzing whether a complex has been formed.