Pharmaceutical Composition Containing Antagonist of EGF Family Ligand as Component

Abstract

The present inventors identified inhibition of a combination of EGFR ligands that serve as targets for inhibition of cancer cell proliferation. More specifically, EREG antagonists and TGFα antagonists were found to be useful as inhibitors of cell growth. The present invention relates to pharmaceutical compositions containing EGF family ligand antagonists as components.

1 . A pharmaceutical composition comprising an EREG antagonist and a TGFα antagonist as active ingredients. 
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein the pharmaceutical composition is an inhibitor of cell proliferation. 
     
     
         3 . The pharmaceutical composition of  claim 1 , wherein the pharmaceutical composition is an anticancer agent. 
     
     
         4 . The pharmaceutical composition of  claim 1 , wherein the EREG antagonist is an anti-EREG antibody. 
     
     
         5 . The pharmaceutical composition of  claim 4 , wherein the anti-EREG antibody is the antibody of any of (a) to (x) below:
 (a) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 61, CDR2 having the amino acid sequence of SEQ ID NO: 62, and CDR3 having the amino acid sequence of SEQ ID NO: 63;   (b) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 64, CDR2 having the amino acid sequence of SEQ ID NO: 65, and CDR3 having the amino acid sequence of SEQ ID NO: 66;   (c) an antibody that comprises the heavy chain of (a) and the light chain of (b);   (d) an antibody that recognizes the same epitope recognized by the antibody of (c);   (e) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 67, CDR2 having the amino acid sequence of SEQ ID NO: 68, and CDR3 having the amino acid sequence of SEQ ID NO: 69;   (f) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 70, CDR2 having the amino acid sequence of SEQ ID NO: 71, and CDR3 having the amino acid sequence of SEQ ID NO: 72;   (g) an antibody that comprises the heavy chain of (e) and the light chain of (f);   (h) an antibody that recognizes the same epitope recognized by the antibody of (g);   (i) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 73, CDR2 having the amino acid sequence of SEQ ID NO: 74, and CDR3 having the amino acid sequence of SEQ ID NO: 75;   (j) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 76, CDR2 having the amino acid sequence of SEQ ID NO: 77, and CDR3 having the amino acid sequence of SEQ ID NO: 78;   (k) an antibody that comprises the heavy chain of (i) and the light chain of (j);   (l) an antibody that recognizes the same epitope recognized by the antibody of (k);   (m) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 79, CDR2 having the amino acid sequence of SEQ ID NO: 80, and CDR3 having the amino acid sequence of SEQ ID NO: 81;   (n) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 82, CDR2 having the amino acid sequence of SEQ ID NO: 83, and CDR3 having the amino acid sequence of SEQ ID NO: 84;   (o) an antibody that comprises the heavy chain of (m) and the light chain of (n);   (p) an antibody that recognizes the same epitope recognized by the antibody of (o);   (q) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 85, CDR2 having the amino acid sequence of SEQ ID NO: 86, and CDR3 having the amino acid sequence of SEQ ID NO: 87;   (r) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 88, CDR2 having the amino acid sequence of SEQ ID NO: 89, and CDR3 having the amino acid sequence of SEQ ID NO: 90;   (s) an antibody that comprises the heavy chain of (q) and the light chain of (r);   (t) an antibody that recognizes the same epitope recognized by the antibody of (s);   (u) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 91, CDR2 having the amino acid sequence of SEQ ID NO: 92, and CDR3 having the amino acid sequence of SEQ ID NO: 93;   (v) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 94, CDR2 having the amino acid sequence of SEQ ID NO: 95, and CDR3 having the amino acid sequence of SEQ ID NO: 96;   (w) an antibody that comprises the heavy chain of (u) and the light chain of (v); and   (x) an antibody that recognizes the same epitope recognized by the antibody of (w).   
     
     
         6 . The pharmaceutical composition of  claim 1 , wherein the TGFα antagonist is an anti-TGFα antibody. 
     
     
         7 . The pharmaceutical composition of  claim 6 , wherein the anti-TGFα antibody is the antibody of any of (a) to (d) below:
 (a) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 47, CDR2 having the amino acid sequence of SEQ ID NO: 48, and CDR3 having the amino acid sequence of SEQ ID NO: 49; 
 (b) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 50, CDR2 having the amino acid sequence of SEQ ID NO: 51, and CDR3 having the amino acid sequence of SEQ ID NO: 52; 
 (c) an antibody that comprises the heavy chain of (a) and the light chain of (b); and 
 (d) an antibody that recognizes the same epitope recognized by the antibody of (c). 
 
     
     
         8 . The pharmaceutical composition of  claim 1 , wherein the EREG antagonist is an anti-EREG antibody, and the TGFα antagonist is an anti-TGFα antibody. 
     
     
         9 . The pharmaceutical composition of  claim 8 , wherein the anti-EREG antibody is the antibody of any of (a) to (x) below:
 (a) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 61, CDR2 having the amino acid sequence of SEQ ID NO: 62, and CDR3 having the amino acid sequence of SEQ ID NO: 63;   (b) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 64, CDR2 having the amino acid sequence of SEQ ID NO: 65, and CDR3 having the amino acid sequence of SEQ ID NO: 66;   (c) an antibody that comprises the heavy chain of (a) and the light chain of (b);   (d) an antibody that recognizes the same epitope recognized by the antibody of (c);   (e) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 67, CDR2 having the amino acid sequence of SEQ ID NO: 68, and CDR3 having the amino acid sequence of SEQ ID NO: 69;   (f) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 70, CDR2 having the amino acid sequence of SEQ ID NO: 71, and CDR3 having the amino acid sequence of SEQ ID NO: 72;   (g) an antibody that comprises the heavy chain of (e) and the light chain of (f);   (h) an antibody that recognizes the same epitope recognized by the antibody of (g);   (i) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 73, CDR2 having the amino acid sequence of SEQ ID NO: 74, and CDR3 having the amino acid sequence of SEQ ID NO: 75;   (j) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 76, CDR2 having the amino acid sequence of SEQ ID NO: 77, and CDR3 having the amino acid sequence of SEQ ID NO: 78;   (k) an antibody that comprises the heavy chain of (i) and the light chain of (j);   (l) an antibody that recognizes the same epitope recognized by the antibody of (k);   (m) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 79, CDR2 having the amino acid sequence of SEQ ID NO: 80, and CDR3 having the amino acid sequence of SEQ ID NO: 81;   (n) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 82, CDR2 having the amino acid sequence of SEQ ID NO: 83, and CDR3 having the amino acid sequence of SEQ ID NO: 84;   (o) an antibody that comprises the heavy chain of (m) and the light chain of (n);   (p) an antibody that recognizes the same epitope recognized by the antibody of (o);   (q) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 85, CDR2 having the amino acid sequence of SEQ ID NO: 86, and CDR3 having the amino acid sequence of SEQ ID NO: 87;   (r) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 88, CDR2 having the amino acid sequence of SEQ ID NO: 89, and CDR3 having the amino acid sequence of SEQ ID NO: 90;   (s) an antibody that comprises the heavy chain of (q) and the light chain of (r);   (t) an antibody that recognizes the same epitope recognized by the antibody of (s);   (u) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 91, CDR2 having the amino acid sequence of SEQ ID NO: 92, and CDR3 having the amino acid sequence of SEQ ID NO: 93;   (v) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 94, CDR2 having the amino acid sequence of SEQ ID NO: 95, and CDR3 having the amino acid sequence of SEQ ID NO: 96;   (w) an antibody that comprises the heavy chain of (u) and the light chain of (v); and   (x) an antibody that recognizes the same epitope recognized by the antibody of (w).   
     
     
         10 . The pharmaceutical composition of  claim 8 , wherein the anti-TGFα antibody is the antibody of any of (a) to (d) below:
 (a) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 47, CDR2 having the amino acid sequence of SEQ ID NO: 48, and CDR3 having the amino acid sequence of SEQ ID NO: 49; 
 (b) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 50, CDR2 having the amino acid sequence of SEQ ID NO: 51, and CDR3 having the amino acid sequence of SEQ ID NO: 52; 
 (c) an antibody that comprises the heavy chain of (a) and the light chain of (b); and 
 (d) an antibody that recognizes the same epitope recognized by the antibody of (c). 
 
     
     
         11 . An antibody that binds to EREG and TGFα. 
     
     
         12 . The antibody of  claim 11 , which has antagonistic activity against EREG and TGFα. 
     
     
         13 . The antibody of  claim 11 , which is the antibody of any one of (a) to (d) below:
 (a) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 10, CDR2 having the amino acid sequence of SEQ ID NO: 12, and CDR3 having the amino acid sequence of SEQ ID NO: 14;   (b) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 16, CDR2 having the amino acid sequence of SEQ ID NO: 18, and CDR3 having the amino acid sequence of SEQ ID NO: 20;   (c) an antibody that comprises the heavy chain of (a) and the light chain of (b); and   (d) an antibody that recognizes the same epitope recognized by the antibody of (c).   
     
     
         14 . An antibody that recognizes the region from Gln at position 56 to Leu at position 102 of SEQ ID NO: 4 (human EREG), and/or the region from Val at position 41 to Leu at position 87 of SEQ ID NO: 26 (human TGFα). 
     
     
         15 . A pharmaceutical composition comprising an EREG- and TGFα-binding substance as an active ingredient. 
     
     
         16 . A pharmaceutical composition comprising an EREG- and TGFα-binding substance as an active ingredient, wherein the EREG- and TGFα-binding substance is the antibody of  claim 11 . 
     
     
         17 . A method of screening for an anticancer agent, which comprises:
 (a) measuring the antagonistic activity of a test substance against EREG;   (b) measuring the antagonistic activity of a test substance against TGFα; and   (c) selecting a test substance that has antagonistic activity against EREG and TGFα.   
     
     
         18 . A method for suppressing cell proliferation, which comprises administering the pharmaceutical composition of  claim 1 . 
     
     
         19 . A method for treating cancer, which comprises administering the pharmaceutical composition of  claim 1  to a subject. 
     
     
         20 . The pharmaceutical composition of  claim 9 , wherein the anti-TGFα antibody is the antibody of any of (a) to (d) below:
 (a) an antibody that comprises a heavy chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 47, CDR2 having the amino acid sequence of SEQ ID NO: 48, and CDR3 having the amino acid sequence of SEQ ID NO: 49; 
 (b) an antibody that comprises a light chain comprising CDR1 having the amino acid sequence of SEQ ID NO: 50, CDR2 having the amino acid sequence of SEQ ID NO: 51, and CDR3 having the amino acid sequence of SEQ ID NO: 52; 
 (c) an antibody that comprises the heavy chain of (a) and the light chain of (b); and 
 (d) an antibody that recognizes the same epitope recognized by the antibody of (c). 
 
     
     
         21 . A pharmaceutical composition comprising an EREG- and TGFα-binding substance as an active ingredient, wherein the EREG- and TGFα-binding substance is the antibody of  claim 13 .