Bispecific anti-vegf/anti-ang-2 antibodies

Abstract

The present invention relates to bispecific antibodies against human VEGF and against human ANG-2, methods for their production, pharmaceutical compositions containing said antibodies, and uses thereof.

1 . A bispecific antibody that binds specifically to human vascular endothelial growth factor (VEGF) and human angiopoietin-2 (ANG-2), said antibody comprising a first antigen-binding site that specifically binds to human VEGF and a second antigen-binding site that specifically binds to human ANG-2. 
     
     
         2 . A bispecific antibody according to  claim 1 , characterized in that
 i) said antigen-binding sites each comprise an antibody heavy chain variable domain and an antibody light chain variable domain;   ii) said first antigen-binding site comprises in the heavy chain variable domain:   a CDR3 region having an amino acid sequence selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 9, SEQ ID NO: 17, and SEQ ID NO: 94;   a CDR2 region having an amino acid sequence selected from the group consisting of: SEQ ID NO: 2, SEQ ID NO: 10, SEQ ID NO: 18, and SEQ ID NO: 95;   and a CDRI region having an amino acid sequence selected from the group consisting of: SEQ ID NO:3, SEQ ID NO: 11, SEQ ID NO: 19, and SEQ ID NO: 96, and   in the light chain variable domain:   a CDR3 region having an amino acid sequence selected from the group consisting of: SEQ ID NO: 4, SEQ ID NO: 12, SEQ ID NO: 20, and SEQ ID NO: 97,   a CDR2 region having an amino acid sequence selected from the group consisting of: SEQ ID NO:5, SEQ ID NO: 13, SEQ ID NO: 21, and SEQ ID NO: 98; and   a CDR1 region having an amino acid sequence selected from the group consisting of: SEQ ID NO:6, SEQ ID NO: 14, SEQ ID NO: 22, and SEQ ID NO: 99; and   iii) said second antigen-binding site comprises in the heavy chain variable domain:   a CDR3 region having an amino acid sequence selected from the group consisting of: SEQ ID NO: 25, SEQ ID NO: 38, SEQ ID NO: 46, SEQ ID NO: 54, SEQ ID NO: 62, SEQ ID NO: 70, SEQ ID NO: 78, and SEQ ID NO: 86;   a CDR2 region having an amino acid sequence selected from the group consisting of: SEQ ID NO: 26, SEQ ID NO: 39, SEQ ID NO: 47, SEQ ID NO: 55, SEQ ID NO: 63, SEQ ID NO: 71, SEQ ID NO: 79, and SEQ ID NO: 87; and   a CDR1 region having an amino acid sequence selected from the group consisting of: SEQ ID NO:27, SEQ ID NO: 40, SEQ ID NO: 48, SEQ ID NO: 56, SEQ ID NO: 64, SEQ ID NO: 72, SEQ ID NO: 80, and SEQ ID NO: 88; and   in the light chain variable domain:   a CDR3 region having an amino acid sequence selected from the group consisting of: SEQ ID NO: 28, SEQ ID NO: 28 with the mutations T92L, H93Q and W94T, SEQ ID NO: 41, SEQ ID NO: 49, SEQ ID NO: 57, SEQ ID NO: 65, SEQ ID NO: 73, SEQ ID NO: 81, and SEQ ID NO: 89;   a CDR2 region having an amino acid sequence selected from the group consisting of: SEQ ID NO:29, SEQ ID NO: 42, SEQ ID NO: 50, SEQ ID NO: 58, SEQ ID NO: 66, SEQ ID NO: 74, SEQ ID NO: 82 and SEQ ID NO: 90; and   a CDR1 region having an amino acid sequence selected from the group consisting of: SEQ ID NO:30, SEQ ID NO: 43, SEQ ID NO: 51, SEQ ID NO: 59, SEQ ID NO: 67, SEQ ID NO: 75, SEQ ID NO: 83, and SEQ ID NO: 91.   
     
     
         3 . A bispecific antibody according to  claim 2 , characterized in that said first antigen-binding site s comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO: 1, a CDR2 region of SEQ ID NO: 2, and a CDR1 region of SEQ ID NO:3, and, in the light chain variable domain, a CDR3 region of SEQ ID NO: 4, a CDR2 region of SEQ ID NO:5, and a CDR1 region of SEQ ID NO:6; and
 said second antigen-binding site comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO: 46, a CDR2 region of SEQ ID NO: 47, and a CDR1 region of SEQ ID NO: 48, and, in the light chain variable domain, a CDR3 region of SEQ ID NO: 49, a CDR2 region of SEQ ID NO: 50, and a CDR1 region of SEQ ID NO: 51.   
     
     
         4 . A bispecific antibody according to  claim 3 , characterized in that
 said first antigen-binding site comprises, as the heavy chain variable domain, SEQ ID NO: 7, and, as the light chain variable domain, SEQ ID NO: 8, and   said second antigen-binding site comprises, as the heavy chain variable domain, SEQ ID NO: 52, and, as the light chain variable domain, SEQ ID NO: 53.   
     
     
         5 . A bispecific antibody according to  claim 2 , characterized in that
 said first antigen-binding site comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO: 1, a CDR2 region of SEQ ID NO: 2, and a CDR1 region of SEQ ID NO:3, and, in the light chain variable domain, a CDR3 region of SEQ ID NO: 4, a CDR2 region of SEQ ID NO:5, and a CDR1 region of SEQ ID NO:6;   said second antigen-binding site comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO: 62, a CDR2 region of SEQ ID NO: 63, and a CDR1 region of SEQ ID NO: 64, and, in the light chain variable domain, a CDR3 region of SEQ ID NO: 65, a CDR2 region of SEQ ID NO: 66, and a CDR1 region of SEQ ID NO: 67.   
     
     
         6 . A bispecific antibody according to  claim 5 , characterized in that
 said first antigen-binding site comprises, as the heavy chain variable domain, SEQ ID NO: 7, and, as the light chain variable domain, SEQ ID NO: 8; and   said second antigen-binding site comprises, as the heavy chain variable domain, SEQ ID NO: 68, and, as the light chain variable domain, SEQ ID NO: 69.   
     
     
         7 . A bispecific antibody according to  claim 2 , characterized in that
 said first antigen-binding site s comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO: 1, a CDR2 region of SEQ ID NO: 2, and a CDR1 region of SEQ ID NO:3, and, in the light chain variable domain, a CDR3 region of SEQ ID NO: 4, a CDR2 region of SEQ ID NO:5, and a CDR1 region of SEQ ID NO:6;   said second antigen-binding site comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO: 78, a CDR2 region of SEQ ID NO: 79, and a CDR1 region of SEQ ID NO: 80, and, in the light chain variable domain, a CDR3 region of SEQ ID NO: 81, a CDR2 region of SEQ ID NO: 82, and a CDR1 region of SEQ ID NO: 83.   
     
     
         8 . A bispecific antibody according to  claim 7 , characterized in that
 said first antigen-binding site comprises, as the heavy chain variable domain, SEQ ID NO: 7, and, as the light chain variable domain, SEQ ID NO: 8; and   said second antigen-binding site comprises, as the heavy chain variable domain SEQ ID NO: 84, and, as the light chain variable domain, SEQ ID NO: 85.   
     
     
         9 . A bispecific antibody according to  claim 1 , characterized in that the ratio of the binding affinities K D  (antigen-binding site specific for VEGF)/K D  (antigen-binding site specific for ANG-2) is 1.0-10.0 
     
     
         10 . A bispecific antibody according to  claim 1 , characterized in that the second antigen binding site that specifically binds to human ANG-2 does not specifically bind to human Angiopoetin 1 (ANG-1). 
     
     
         11 . A bispecific antibody according to  claim 1 , characterized in that said antibody is bivalent, trivalent or tetravalent 
     
     
         12 . A pharmaceutical composition comprising an antibody according to  claim 1 . 
     
     
         13 . A method of treatment of a patient suffering from cancer comprising the step of administering an antibody according to  claim 1  to a patient in the need of such treatment. 
     
     
         14 . A method of treatment of a patient suffering from a vascular disease comprising the step of administering an antibody according to  claim 1  to a patient in the need of such treatment. 
     
     
         15 . A nucleic acid encoding a bispecific antibody according to  claim 1 . 
     
     
         16 . An expression vector which comprises the nucleic acid according  claim 15  and is capable of expressing said nucleic acid in a prokaryotic or eukaryotic host cell. 
     
     
         17 . A prokaryotic or eukaryotic host cell comprising a vector according to  claim 16 . 
     
     
         18 . A method for the production of a bispecific antibody according to  claim 1 , comprising expressing a nucleic acid according to  claim 15  in a prokaryotic or eukaryotic host cell and recovering said bispecific antibody from said cell or the cell culture supernatant. 
     
     
         19 . An antibody obtained by the recombinant method of  claim 18 .