Multispecific antibodies against cd40 and cd137

Abstract

Multispecific antibodies binding to human CD40 and human CD137, methods for preparing such multispecific antibodies, and methods of using such multispecific antibodies for therapeutic or other purposes.

1 . A multispecific antibody comprising
 (I) a first antigen-binding region binding to human CD40, wherein said first antigen-binding region comprises heavy and light chain variable region CDR1, CDR2, and CDR3 selected from the group consisting of:   a) heavy chain variable region CDR1, CDR2 and CDR3 having the amino acid sequences set forth in SEQ ID NOs:1, 2 and 3, respectively, and light chain variable region CDR1, CDR2 and CDR3 having the amino acid sequences set forth in SEQ ID NOs:4, YTS and 5, respectively;   b) heavy and light chain variable region CDR1, CDR2 and CDR3 according to a) having a total of one to twelve mutations; and   c) heavy and light chain variable region CDR1, CDR2 and CDR3 of an antibody which (i) competes for human CD40 binding with an antibody comprising heavy and light chain variable region CDR1, CDR2 and CDR3 according to a) orb) and/or (ii) has the specificity for CD40 of an antibody comprising heavy and light chain variable region CDR1, CDR2 and CDR3 according to a) or b),
 and 
   (II) a second antigen-binding region binding to human CD137.   
     
     
         2 . (canceled) 
     
     
         3 . The multispecific antibody of  claim 1 , wherein said first antigen-binding region comprises heavy chain variable region CDR1, CDR2 and CDR3 having the sequences set forth in SEQ ID NOs:1, 2 and 3, respectively, and light chain variable region CDR1, CDR2 and CDR3 having the sequences set forth in SEQ ID NOs:4, YTS and 5, respectively. 
     
     
         4 - 5 . (canceled) 
     
     
         6 . The multispecific antibody according to of  claim 1 , wherein said first antigen-binding region comprises a first heavy chain variable (VH) sequence and a first light chain variable (VL) sequence, said second antigen-binding region comprises a second VH sequence and a second VL sequence, wherein said VH sequence of the first antigen-binding region comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% identity to SEQ ID NO:117; and wherein said VL sequence of the first antigen-binding region comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% identity to SEQ ID NOS:121. 
     
     
         7 - 11 . (canceled) 
     
     
         12 . The multispecific antibody according to of  claim 1 , wherein said second antigen-binding region comprises heavy and light chain variable region CDR1, CDR2 and CDR3 selected from the group consisting of:
 a) heavy chain variable region CDR1, CDR2 and CDR3 having the sequences set forth in SEQ ID NOs:64, 65 and 66, respectively, and light chain variable region CDR1, CDR2 and CDR3 having the sequences set forth in SEQ ID NOs:67, GAS and 68, respectively, and   b) heavy and light chain variable region CDR1, CDR2 and CDR3 according to a) having a total of one to twelve mutations.   c) heavy and light chain variable region CDR1, CDR2 and CDR3 of an antibody which (i) competes for human CD137 binding with an antibody comprising heavy and light chain variable region CDR1, CDR2 and CDR3 according to a) orb) and/or (ii) has the specificity for CD137 of an antibody comprising heavy and light chain variable region CDR1, CDR2 and CDR3 according to a), or b).   
     
     
         13 - 14 . (canceled) 
     
     
         15 . The multispecific antibody according to of  claim 1 , wherein said VH and said VL sequence of the second antigen-binding region comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% identity to SEQ ID NO:123 and SEQ ID NO:127, respectively. 
     
     
         16 - 18 . (canceled) 
     
     
         19 . The multispecific antibody according to of  claim 1 , wherein said first antigen-binding region comprises a first heavy chain variable (VH) sequence and a first light chain variable (VL) sequence, said second antigen-binding region comprises a second VH sequence and a second VL sequence, wherein said VH and VL sequence of the second antigen-binding region comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97% or at least 99% identity to SEQ ID NO:41; and SEQ ID NO:42, respectively. 
     
     
         20 - 21 . (canceled) 
     
     
         22 . The multispecific antibody of  claim 1 , wherein said antibody comprises (I) a first binding arm comprising said first antigen-binding region, and (II) a second binding arm comprising said second antigen-binding region. 
     
     
         23 . (canceled) 
     
     
         24 . The multispecific antibody of  claim 22 , wherein (I) said first binding arm comprises a first heavy chain comprising a first heavy chain variable (VH) sequence and a first heavy chain constant (CH) sequence, and a first light chain comprising a first light chain variable (VL) sequence, and (II) said second binding arm comprises a second heavy chain comprising a second heavy chain variable (VH) sequence and a second heavy chain constant (CH) sequence, and a second light chain comprising a second light chain variable (VL) sequence. 
     
     
         25 - 39 . (canceled) 
     
     
         40 . The multispecific of  claim 24 , 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 according to EU numbering 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 according to EU numbering has been substituted, and wherein said first and said second heavy chains are not substituted in the same positions. 
     
     
         41 . (canceled) 
     
     
         42 . The multispecific antibody according to of  claim 1 , wherein said antibody comprises a first and a second heavy chain, and wherein said antibody induces Fc-mediated effector function to a lesser extent compared to a multispecific antibody comprising the same first and second antigen-binding regions and two heavy chains comprising human IgG1 hinge, CH2 and CH3 regions. 
     
     
         43 - 46 . (canceled) 
     
     
         47 . The multispecific antibody according to of  claim 1 , wherein said antibody comprises a first and a second heavy chain, wherein in at least one of said first and second heavy chains one or more amino acids in the positions corresponding to positions L234, L235, D265, N297, and P331 in a human IgG1 heavy chain according to EU numbering, are not L, L, D, N, and P, respectively. 
     
     
         48 - 49 . (canceled) 
     
     
         50 . The multispecific antibody according to claim  49 , wherein the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain according to EU numbering of both the first heavy chain and the second heavy chain are F, E, and A, respectively, and wherein (i) the position corresponding to F405 in a human IgG1 heavy chain according to EU numbering of the first heavy chain is L, and the position corresponding to K409 in a human IgG1 heavy chain according to EU numbering of the second heavy chain is R, or (ii) the position corresponding to K409 in a human IgG1 heavy chain according to EU numbering of the first heavy chain is R, and the position corresponding to F405 in a human IgG1 heavy chain according to EU numbering of the second heavy chain is L. 
     
     
         51 - 63 . (canceled) 
     
     
         64 . A nucleic acid encoding the multispecific antibody of  claim 1 . 
     
     
         65 . An expression vector comprising the nucleic acid of  claim 64 . 
     
     
         66 . A host cell comprising the nucleic acid according to of  claim 64 . 
     
     
         67 . (canceled) 
     
     
         68 . A composition comprising the multispecific antibody of  claim 1 . 
     
     
         69 - 75 . (canceled) 
     
     
         76 . A method of treatment of a disease comprising administering the multispecific antibody of  claim 1 . 
     
     
         77 . (canceled) 
     
     
         78 . The method of  claim 76 , wherein the method comprises further administration of one or more further therapeutic agent, such as a chemotherapeutic agent. 
     
     
         79 . A method for producing the multispecific antibody according to  claim 1 , wherein the multispecific antibody is a bispecific antibody, comprising the steps of
 a) providing a first antibody comprising an Fc region, said Fc region comprising a first CH3 region;   b) providing a second antibody comprising a second Fc region, said Fc region comprising a second CH3 region,
 wherein the first antibody is a CD40 antibody comprising two first antigen-binding regions according to  claim 1 , and the second antibody is a CD137 antibody comprising two second antigen-binding regions according to  claim 1 , or vice versa; and wherein the sequences of said first and second CH3 regions are different and are such that the heterodimeric interaction between said first and second CH3 regions is stronger than each of the homodimeric interactions of said first and second CH3 regions; 
   c) incubating said first antibody together with said second antibody under reducing conditions; and   d) obtaining said bispecific CD40×CD137 antibody.   
     
     
         80 . (canceled) 
     
     
         81 . A kit for detecting cross-linking between CD40- and CD137-expressing cells, in a sample obtained from a patient such as a blood sample, lymph node sample or bone marrow sample, comprising
 i) the multispecific antibody of  claim 1 ; and   ii) instructions for use of said kit.   
     
     
         82 . An anti-idiotypic antibody which binds to the second or first and second antigen-binding region as defined in  claim 1 .