Notch binding agents and antagonists and methods of use thereof

Abstract

The present invention relates to Notch-binding agents and Notch antagonists and methods of using the agents and/or antagonists for treating diseases such as cancer. The present invention provides antibodies that specifically bind to a non-ligand binding region of the extracellular domain of one or more human Notch receptor, such as Notch2 and/or Notch3, and inhibit tumor growth. The present invention further provides methods of treating cancer, the methods comprising administering a therapeutically effective amount of an antibody that specifically binds to a non-ligand binding region of the extracellular domain of a human Notch receptor protein and inhibits tumor growth.

1 - 89 . (canceled) 
     
     
         90 . An isolated antibody that competes with a reference antibody for specific binding to a non-ligand binding region of the extracellular domain of human Notch2 and/or Notch3, wherein the reference antibody comprises:
 (a) a heavy chain CDR1 comprising SSSGMS (SEQ ID NO:5), a heavy chain CDR2 comprising VIASSGSNTYYADSVKG (SEQ ID NO:6), and a heavy chain CDR3 comprising SIFYTT (SEQ ID NO:51); and a light chain CDR1 comprising RASQSVRSNYLA (SEQ ID NO:8), a light chain CDR2 comprising GASSRAT (SEQ ID NO:9), and a light chain CDR3 comprising QQYSNFPI (SEQ ID NO:10); or   (b) a heavy chain CDR1 comprising SSSGMS (SEQ ID NO:5), a heavy chain CDR2 comprising VIASSGSNTYYADSVKG (SEQ ID NO:6), and a heavy chain CDR3 comprising GIFFAI (SEQ ID NO:7); and a light chain CDR1 comprising RASQSVRSNYLA (SEQ ID NO:8), a light chain CDR2 comprising GASSRAT (SEQ ID NO:9), and a light chain CDR3 comprising QQYSNFPI (SEQ ID NO:10).   
     
     
         91 . The antibody of  claim 90 , wherein the reference antibody comprises:
 a heavy chain CDR1 comprising SSSGMS (SEQ ID NO:5), a heavy chain CDR2 comprising VIASSGSNTYYADSVKG (SEQ ID NO:6), and a heavy chain CDR3 comprising SIFYTT (SEQ ID NO:51); and a light chain CDR1 comprising RASQSVRSNYLA (SEQ ID NO:8), a light chain CDR2 comprising GASSRAT (SEQ ID NO:9), and a light chain CDR3 comprising QQYSNFPI (SEQ ID NO:10).   
     
     
         92 . The antibody of  claim 90 , wherein the reference antibody comprises:
 (a) a heavy chain variable region having at least about 90% sequence identity to SEQ ID NO:50 and a light chain variable region having at least about 90% sequence identity to SEQ ID NO:13; or   (b) a heavy chain variable region having at least about 90% sequence identity to SEQ ID NO:14 and a light chain variable region having at least about 90% sequence identity to SEQ ID NO:13.   
     
     
         93 . The antibody of  claim 90 , wherein the reference antibody comprises:
 (a) a heavy chain variable region having at least about 95% sequence identity to SEQ ID NO:50 and a light chain variable region having at least about 95% sequence identity to SEQ ID NO:13; or   (b) a heavy chain variable region having at least about 95% sequence identity to SEQ ID NO:14 and a light chain variable region having at least about 95% sequence identity to SEQ ID NO:13.   
     
     
         94 . The antibody of  claim 90 , wherein the reference antibody comprises a heavy chain variable region comprising SEQ ID NO:50 and a light chain variable region comprising SEQ ID NO:13. 
     
     
         95 . The antibody of  claim 90 , wherein the reference antibody comprises a heavy chain variable region comprising SEQ ID NO:14 and a light chain variable region comprising SEQ ID NO:13. 
     
     
         96 . The antibody of  claim 90 , which is a monoclonal antibody, a chimeric antibody, a humanized antibody, a human antibody, a bispecific antibody, or an antibody fragment. 
     
     
         97 . A pharmaceutical composition comprising the antibody of  claim 90 . 
     
     
         98 . An isolated antibody that competes with a reference antibody for specific binding to epidermal growth factor (EGF) repeat 10 of human Notch2 and EGF repeat 9 of human Notch3. 
     
     
         99 . The antibody of  claim 98 , wherein the reference antibody binds at least part of the sequence HKGAL (SEQ ID NO:28) within EGF repeat 10 of human Notch2. 
     
     
         100 . The antibody of  claim 99 , wherein the reference antibody binds at least part of the sequence HEDAI (SEQ ID NO:29) within EGF repeat 9 of human Notch3. 
     
     
         101 . The antibody of  claim 98 , wherein the reference antibody binds at least part of the sequence HEDAI (SEQ ID NO:29) within EGF repeat 9 of human Notch3. 
     
     
         102 . The antibody of  claim 98 , which is a monoclonal antibody, a chimeric antibody, a humanized antibody, a human antibody, a bispecific antibody, or an antibody fragment. 
     
     
         103 . A pharmaceutical composition comprising the antibody of  claim 98 . 
     
     
         104 . A method of treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of an isolated antibody that specifically binds a non-ligand binding region of the extracellular domain of human Notch2 and/or Notch3, wherein the antibody competes for binding with a reference antibody comprising:
 (a) a heavy chain CDR1 comprising SSSGMS (SEQ ID NO:5), a heavy chain CDR2 comprising VIASSGSNTYYADSVKG (SEQ ID NO:6), and a heavy chain CDR3 comprising SIFYTT (SEQ ID NO:51); and a light chain CDR1 comprising RASQSVRSNYLA (SEQ ID NO:8), a light chain CDR2 comprising GASSRAT (SEQ ID NO:9), and a light chain CDR3 comprising QQYSNFPI (SEQ ID NO:10); or   (b) a heavy chain CDR1 comprising SSSGMS (SEQ ID NO:5), a heavy chain CDR2 comprising VIASSGSNTYYADSVKG (SEQ ID NO:6), and a heavy chain CDR3 comprising GIFFAI (SEQ ID NO:7); and a light chain CDR1 comprising RASQSVRSNYLA (SEQ ID NO:8), a light chain CDR2 comprising GASSRAT (SEQ ID NO:9), and a light chain CDR3 comprising QQYSNFPI (SEQ ID NO:10).   
     
     
         105 . The method of  claim 104 , further comprising administering to the subject at least one additional cancer treatment. 
     
     
         106 . The method of  claim 105 , wherein at least one additional cancer treatment is a chemotherapeutic agent. 
     
     
         107 . A method of treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of an isolated antibody that specifically human Notch2 and Notch3, wherein the antibody competes for binding with a reference antibody that binds epidermal growth factor (EGF) repeat 10 of human Notch2 and EGF repeat 9 of human Notch 3. 
     
     
         108 . The method of  claim 107 , further comprising administering to the subject at least one additional cancer treatment. 
     
     
         109 . The method of  claim 108 , wherein at least one additional cancer treatment is a chemotherapeutic agent.