Bispecific anti-cd37 antibodies, monoclonal anti-cd37 antibodies and methods of use thereof

Abstract

CD37-specific bispecific antibody molecules binding to different epitopes of the human CD37 antigen which bispecific antibody molecules have enhanced Fc-Fc interactions upon binding to CD37 on the cell surface. The invention also relates to the monoclonal parental antibodies from which the first or the second binding region of the bispecific antibody molecules is obtained. The invention also relates to pharmaceutical compositions containing these molecules and the treatment of cancer and other diseases using these compositions.

1 . A bispecific antibody comprising a first antigen binding region and a second antigen binding region, both of which bind to human CD37 having the sequence set forth in SEQ ID NO: 62, and first and second Fc regions of a human immunoglobulin, wherein the first and second antigen binding regions bind to different epitopes on human CD37, and wherein the first and second Fc regions comprise one or more amino acid mutations which enhance Fc-Fc interactions between two or more of the bispecific antibodies upon binding to a membrane-bound target compared to the Fc-Fc interactions between the bispecific antibodies which lack said mutation(s). 
     
     
         2 . The bispecific antibody of  claim 1 , wherein said first antigen binding region competes for binding to human CD37 with or binds to the same epitope on human CD37 as a CD37 antibody comprising:
 (a) a variable heavy chain (VH) region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 16, 17, and 18, respectively, and a variable light chain (VL) region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 20, KAS, and SEQ ID NO: 21, respectively, or   (b) a variable heavy chain (VH) region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 9, 10, and 11, respectively, and a variable light chain (VL) region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 13, AAS, and SEQ ID NO: 14, respectively.   
     
     
         3 . (canceled) 
     
     
         4 . The bispecific antibody of  claim 1 , wherein said first antigen binding region comprises:
 (a) a variable heavy chain (VH) region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 16, 17, and 18, respectively, and a variable light chain (VL) region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 20, KAS, and SEQ ID NO: 21, respectively, or   (b) a variable heavy chain (VH) region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 9, 10, and 11, respectively, and a variable light chain (VL) region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 13, AAS, and SEQ ID NO: 14, respectively.   
     
     
         5 . The bispecific antibody of  claim 1 , wherein said first antigen binding region comprises a variable heavy chain (VH) region and a variable light chain (VL) region selected from the group consisting of:
 (i) a variable heavy chain (VH) region comprising the amino acid sequence set forth in SEQ ID NO: 15 and a variable light chain (VL) region comprising the amino acid sequence set forth in SEQ ID NO: 19,   (ii) a VH region comprising an amino acid sequence having at least 90% identity to a VH region comprising the amino acid sequence set forth in SEQ ID NO: 15, and a VL region comprising an amino acid sequence having at least 90% identity to a VL region comprising the amino acid sequences set forth in SEQ ID NO: 19,   (iii) a VH region comprising the amino acid sequence set forth in SEQ ID NO: 8 and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 12, and   (iv) a VH region comprising an amino acid sequence having at least 90% identity to a VH region comprising the amino acid sequence set forth in SEQ ID NO: 8, and a VL region comprising an amino acid sequence having at least 90% identity to a VL region comprising the amino acid sequences set forth in SEQ ID NO: 12.   
     
     
         6 . The bispecific antibody of  claim 1 , wherein said first antigen binding region binds to a functional epitope comprising one or more amino acids selected from the group consisting of: Y182, D189, T191, 1192, D194, K195, V196, 1197 and P199 of human CD37. 
     
     
         7 - 10 . (canceled) 
     
     
         11 . The bispecific antibody of  claim 1 , wherein said second antigen binding region competes for binding to human CD37 with or binds to the same epitope on human CD37 as a CD37 antibody comprising a variable heavy chain (VH) region and variable light chain (VL) region selected from the group consisting of:
 (i) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 23, 24, and 25, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 27, YAS, and SEQ ID NO: 28, respectively;   (ii) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 23, 24, and 25, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 27, YAS, and SEQ ID NO: 31, respectively;   (iii) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 2, 3, and 4, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 6, EAS, and SEQ ID NO: 7, respectively;   (iv) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 40, 41, and 42, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 44, FAK, and SEQ ID NO: 45, respectively; and   (v) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 47, 48, and 49, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 51, VAT, and SEQ ID NO: 52, respectively.   
     
     
         12 . (canceled) 
     
     
         13 . The bispecific antibody of  claim 1 , wherein said second antigen binding region binds to a functional epitope comprising one or more amino acids selected from the group consisting of: E124, F62, Q163, V164, L165 and H175 of human CD37. 
     
     
         14 . The bispecific antibody of  claim 1 , wherein said second antigen binding region comprises a variable heavy chain (VH) region and a variable light chain (VL) region selected from the group consisting of:
 (i) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 23, 24, and 25, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 27, YAS, and SEQ ID NO: 28, respectively;   (ii) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 23, 24, and 25, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 27, YAS, and SEQ ID NO: 31, respectively;   (iii) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 2, 3, and 4, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 6, EAS, and SEQ ID NO: 7, respectively;   (iv) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 40, 41, and 42, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 44, FAK, and SEQ ID NO: 45, respectively; and   (v) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 47, 48, and 49, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 51, VAT, and SEQ ID NO: 52, respectively.   
     
     
         15 . The bispecific antibody of  claim 1 , wherein said second antigen binding region comprises a variable heavy chain (VH) region and variable light chain (VL) region selected from the group consisting of:
 (i) a VH region comprising the amino acid sequence set forth in SEQ ID NO: 22 and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 26;   (ii) a VH region comprising the amino acid sequence set forth in SEQ ID NO: 22 and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 29;   (iii) a VH region comprising the amino acid sequence set forth in SEQ ID NO: 1 and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 5;   (iv) a VH region comprising the amino acid sequence set forth in SEQ ID NO: 39 and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 43;   (v) a VH region comprising the amino acid sequence set forth in SEQ ID NO: 46 and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 50; and   (vi) a VH region comprising an amino acid sequence having at least 90% identity to a VH region comprising the amino acid sequence set forth in any one of (i) to (v), and a VL region comprising an amino acid sequence having at least 90% identity to a VL region comprising the amino acid sequence set forth in any one of (i) to (v).   
     
     
         16 . (canceled) 
     
     
         17 . The bispecific antibody of  claim 1 , wherein the one or more mutations which enhance Fc-Fc interactions in said first and second Fc regions correspond to amino acid positions 430, 440 and 345 in human IgG1, wherein the numbering of positions is according to the EU numbering system. 
     
     
         18 . The bispecific antibody of  claim 17 , comprising at least one substitution in said first and second Fc regions selected from the group consisting of: E430G, E345K, E430S, E430F, E430T, E345Q, E345R, E345Y, S440Y and S440W. 
     
     
         19 - 20 . (canceled) 
     
     
         21 . The bispecific antibody of  claim 1 , wherein the bispecific antibody is an IgG1, IgG2, IgG3 or IgG4 isotype or a combination thereof. 
     
     
         22 . (canceled) 
     
     
         23 . The bispecific antibody of  claim 1 , wherein the bispecific antibody is human, humanized or chimeric. 
     
     
         24 . (canceled) 
     
     
         25 . The bispecific antibody of  claim 1 , wherein
 (i) the first Fc region comprises a mutation corresponding to F405L in human IgG1 and the second Fc region comprises a mutation corresponding to K409R in human IgG1, or   (ii) the second Fc region comprises a mutation corresponding to F405L in human IgG1 and the first Fc region comprises a mutation corresponding to K409R in human IgG1,   when the numbering of positions is according to the EU numbering system.   
     
     
         26 - 27 . (canceled) 
     
     
         28 . An anti-CD37 antibody which binds to human CD37, wherein the antibody comprises a variable heavy chain (VH) region and a variable light chain (VL) region selected from the group consisting of:
 (i) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 16, 17, and 18, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 20, KAS, and SEQ ID NO: 2, respectively;   (ii) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 9, 10, and 11, respectively, and a variable light chain (VL) region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 113, AAS, and SEQ ID NO: 14, respectively;   (iii) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 23, 24, and 25, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 27, YAS, and SEQ ID NO: 28, respectively;   (iv) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 23, 24, and 25, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 27, YAS, and SEQ ID NO: 31, respectively;   (v) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 2, 3, and 4, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 6, EAS, and SEQ ID NO: 7, respectively;   (vi) a variable heavy chain (VH) region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 40, 41, and 42, respectively, and a variable light chain (VL) region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 44, FAK, and SEQ ID NO: 45, respectively, and   (vii) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 47, 48, and 49, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 51, VAT, and SEQ ID NO: 52, respectively,   wherein the antibody of any one of (i)-(vii) optionally comprises an Fc region comprising at least one amino acid substitution selected from the group consisting of: E430G, E345K, E430S, E430F, E430T, E345Q, E345R, E345Y, S440Y and S440W; and   wherein optionally the Fc region further comprises a K409R or F405L mutation.   
     
     
         29 - 35 . (canceled) 
     
     
         36 . The antibody of  claim 28 , wherein the antibody is an IgG1, IgG2, IgG3 or IgG4 isotype. 
     
     
         37 . The antibody of  claim 28 , wherein the antibody is human, humanized or chimeric. 
     
     
         38 . (canceled) 
     
     
         39 . A pharmaceutical composition comprising the antibody of  claim 28  or a bispecific antibody comprising the antigen-binding region of the antibody, and a pharmaceutically acceptable carrier. 
     
     
         40 . (canceled) 
     
     
         41 . A method of treating cancer, an autoimmune disease, or an inflammatory disorder comprising administering to an individual in need thereof a therapeutically effective amount of the antibody of  claim 28 , a bispecific antibody comprising the antigen-binding region of the antibody, or a composition comprising the antibody or bispecific antibody. 
     
     
         42 . The method of  claim 41 , wherein the cancer, autoimmune disease, or inflammatory disorder is selected from the group consisting of: allergy, transplantation rejection or a B-cell malignancy, such as non-Hodgkin lymphoma (NHL), chronic lymphocytic leukemia (CLL), follicular lymphoma (FL), mantle cell lymphoma (MCL), plasma cell leukemia (PCL), diffuse large B-cell lymphoma (DLBCL), or acute lymphoblastic leukemia (ALL). 
     
     
         43 . The method of  claim 41 , further comprising administering one or more further therapeutic agents such as doxorubicin, cisplatin, bleomycin, carmustine, cyclophosphamide, chlorambucil, bendamustine, vincristine, fludarabine, ibrutinib and an anti-CD20 antibody such as rituximab or ofatumumab. 
     
     
         44 . (canceled) 
     
     
         45 . The method of  claim 43 , wherein the anti-CD20 antibody comprises a variable heavy chain (VH) region and a variable light chain (VL) region selected from the group consisting of:
 (i) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 75, 76, and 77, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 79, DAS, and SEQ ID NO: 80, respectively;   (ii) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 82, 83, and 84, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 85, DAS, and SEQ ID NO: 86, respectively;   (iii) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 95, 96, and 97, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 99, ATS, and SEQ ID NO: 100, respectively;   (iv) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 88, 89, and 90, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 92, DAS, and SEQ ID NO: 93, respectively; and   (v) a VH region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NOs: 102, 103, and 104, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences comprising the amino acid sequences set forth in SEQ ID NO: 106, QMS, and SEQ ID NO: 107, respectively.   
     
     
         46 - 50 . (canceled) 
     
     
         51 . A method of inducing cell death, or inhibiting growth and/or proliferation of a tumor cell expressing CD37, comprising administering to an individual in need thereof an effective amount of the antibody of  claim 28 , a bispecific antibody comprising the antigen-binding region of the antibody, or a composition comprising the antibody or bispecific antibody. 
     
     
         52 - 54 . (canceled) 
     
     
         55 . A nucleic acid construct encoding one or more sequences selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 6a, 7, 8, 9, 10, 11, 12, 13, 13a, 14, 15, 16, 17, 18, 19, 20, 20a, 21, 22, 23, 24, 25, 26, 27, 27a, 28, 29, 30, 30a and 31 as set out in Table 1. 
     
     
         56 . A nucleic acid construct encoding the antibody of  claim 28 , or a bispecific antibody comprising the antigen binding region of the antibody. 
     
     
         57 . An expression vector comprising one or more nucleic acid constructs of  claim 55 . 
     
     
         58 . A host cell comprising the expression vector of  claim 57 . 
     
     
         59 . (canceled) 
     
     
         60 . An anti-idiotypic antibody, which binds to the antibody of  claim 28 . 
     
     
         61 . An in vitro method for detecting the presence of a human CD37 antigen or a cell expressing human CD37 in a sample, said method comprising:
 (i) contacting the sample with the antibody of  claim 28 , or a bispecific antibody comprising the antigen-binding region of the antibody, under conditions that allow for formation of a complex between the antibody or the bispecific antibody and CD37; and   (ii) detecting the formation of a complex.   
     
     
         62 . An in vivo method for detecting the presence of a human CD37 antigen, or a cell expressing human CD37 in a subject, said method comprising:
 (i) administering the antibody of  claim 28 , or a bispecific antibody comprising the antigen-binding region of the antibody, under conditions that allow for formation of a complex between the antibody or the bispecific antibody and CD37; and   (ii) detecting the formed complex.