Met-binding agents and uses thereof

Abstract

The present invention relates to binding agents that specifically bind human MET, binding agents that specifically bind one or more components of the WNT pathway, bispecific agents that bind both human MET and one or more components of the WNT pathway, and methods of using the agents for treating diseases such as cancer.

1 - 2 . (canceled) 
     
     
         3 . A method of inhibiting Wnt signaling in a cell, comprising contacting the cell with an effective amount of a bispecific agent, wherein the bispecific agent comprises:
 a) a first binding site comprising an antigen-binding site of an antibody that specifically binds human MET, wherein the antigen-binding site comprises a heavy chain CDR1 comprising ASYAWS (SEQ ID NO:1), a heavy chain CDR2 comprising YISYSGGTDYNPSLKS (SEQ ID NO:2), and a heavy chain CDR3 comprising KGAY (SEQ ID NO:3); and a light chain CDR1 comprising SASSSVSSSYLY (SEQ ID NO:4), a light chain CDR2 comprising STSNLAS (SEQ ID NO:5), and a light chain CDR3 comprising HQWSSYPYT (SEQ ID NO:6); and   b) a second binding site that specifically binds one or more components of the WNT pathway, wherein the second binding site comprises a soluble human frizzled 8 (FZD8) receptor.   
     
     
         4 . The method of  claim 3 , wherein the second binding site of the bispecific agent comprises the Fri domain of human FZD8. 
     
     
         5 . The method of  claim 4 , wherein the Fri domain of human FZD8 comprises SEQ ID NO:28, SEQ ID NO:29, or SEQ ID NO:39. 
     
     
         6 . The method of  claim 4 , wherein the Fri domain of human FZD8 is linked to a heterologous polypeptide. 
     
     
         7 . The method of  claim 6 , wherein the heterologous polypeptide comprises a human Fc region. 
     
     
         8 . The method of  claim 3 , wherein the soluble FZD8 receptor comprises SEQ ID NO:56. 
     
     
         9 . The method of  claim 3 , wherein the first binding site of the bispecific agent comprises a heavy chain variable region comprising SEQ ID NO:7 and a light chain variable region comprising SEQ ID NO:8. 
     
     
         10 . The method of  claim 3 , wherein the first binding site of the bispecific agent comprises SEQ ID NO: 13 and SEQ ID NO: 14 and the second binding site comprises SEQ ID NO:56. 
     
     
         11 . The method of  claim 3 , wherein the first binding site of the bispecific agent comprises a heavy chain variable region encoded by the plasmid deposited with ATCC designated PTA-13609 and a light chain variable region encoded by the plasmid deposited with ATCC designated PTA-13610; and the second binding site comprises a polypeptide encoded by the plasmid deposited with ATCC designated PTA-13611. 
     
     
         12 . The method of  claim 3 , wherein the bispecific agent comprises a first human IgG2 constant region with amino acid substitutions at positions corresponding to positions 249 and 288 of SEQ ID NO:75, wherein the amino acids are replaced with glutamate or aspartate, and a second human IgG2 constant region with amino acid substitutions at positions corresponding to positions 236 and 278 of SEQ ID NO:75, wherein the amino acids are replaced with lysine. 
     
     
         13 . The method of  claim 3 , wherein the cell is a tumor cell. 
     
     
         14 . The method of  claim 3 , wherein the Wnt signaling is canonical Wnt signaling. 
     
     
         15 . The method of  claim 3 , wherein the method comprises contacting the cell with at least one additional therapeutic agent. 
     
     
         16 . A method of inhibiting MET activation in a cell, comprising contacting the cell with an effective amount of a bispecific agent, wherein the bispecific agent comprises:
 a) a first binding site comprising an antigen-binding site of an antibody that specifically binds human MET, wherein the antigen-binding site comprises a heavy chain CDR1 comprising ASYAWS (SEQ ID NO:1), a heavy chain CDR2 comprising YISYSGGTDYNPSLKS (SEQ ID NO:2), and a heavy chain CDR3 comprising KGAY (SEQ ID NO:3); and a light chain CDR1 comprising SASSSVSSSYLY (SEQ ID NO:4), a light chain CDR2 comprising STSNLAS (SEQ ID NO:5), and a light chain CDR3 comprising HQWSSYPYT (SEQ ID NO:6); and   b) a second binding site that specifically binds one or more components of the WNT pathway, wherein the second binding site comprises a soluble human frizzled 8 (FZD8) receptor.   
     
     
         17 . The method of  claim 16 , wherein the second binding site of the bispecific agent comprises the Fri domain of human FZD8. 
     
     
         18 . The method of  claim 17 , wherein the Fri domain of human FZD8 comprises SEQ ID NO:28, SEQ ID NO:29, or SEQ ID NO:39. 
     
     
         19 . The method of  claim 17 , wherein the Fri domain of human FZD8 is linked to a heterologous polypeptide. 
     
     
         20 . The method of  claim 19 , wherein the heterologous polypeptide comprises a human Fc region. 
     
     
         21 . The method of  claim 16 , wherein the soluble FZD8 receptor comprises SEQ ID NO:56. 
     
     
         22 . The method of  claim 16 , wherein the first binding site of the bispecific agent comprises a heavy chain variable region comprising SEQ ID NO:7 and a light chain variable region comprising SEQ ID NO:8. 
     
     
         23 . The method of  claim 16 , wherein the first binding site of the bispecific agent comprises SEQ ID NO:13 and SEQ ID NO:14 and the second binding site comprises SEQ ID NO:56. 
     
     
         24 . The method of  claim 16 , wherein the first binding site of the bispecific agent comprises a heavy chain variable region encoded by the plasmid deposited with ATCC designated PTA-13609 and a light chain variable region encoded by the plasmid deposited with ATCC designated PTA-13610; and the second binding site comprises a polypeptide encoded by the plasmid deposited with ATCC designated PTA-13611. 
     
     
         25 . The method of  claim 16 , wherein the cell is a tumor cell. 
     
     
         26 . A polynucleotide comprising a sequence selected from the group consisting of: SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:89, and SEQ ID NO:90.