Notch1 receptor binding agents and methods of use thereof

Abstract

The present invention relates to compositions and methods for characterizing, diagnosing, and treating cancer. In particular the invention provides the means and methods for the diagnosis, characterization, prognosis and treatment of cancer and specifically targeting cancer stem cells. The present invention provides an antibody that specifically binds to a non-ligand binding membrane proximal region of the extracellular domain of a human Notch receptor and inhibits tumor growth. The present invention further provides a method of treating cancer, the method comprising administering a therapeutically effective amount of an antibody that specifically binds to a non-ligand binding membrane proximal region of the extracellular domain of a human Notch receptor protein and inhibits tumor growth.

1 - 84 . (canceled) 
     
     
         85 . A method of treating colorectal cancer in a subject, comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds human Notch1, wherein the antibody comprises:
 (a) a heavy chain CDR1 comprising RGYWIE (SEQ ID NO:15), a heavy chain CDR2 comprising QILPGTGRTNYNEKFKG (SEQ ID NO:16), and a heavy chain CDR3 comprising FDGNYGYYAMDY (SEQ ID NO:17); and   (b) a light chain CDR1 comprising RSSTGAVTTSNYAN (SEQ ID NO:18), a light chain CDR2 comprising GTNNRAP (SEQ ID NO:19), and a light chain CDR3 comprising ALWYSNHWVFGGGTKL (SEQ ID NO:20).   
     
     
         86 . The method of  claim 85 , wherein the antibody binds to a non-ligand binding membrane proximal region of an extracellular domain of human Notch1 receptor. 
     
     
         87 . The method of  claim 85 , wherein the antibody comprises a heavy chain variable region having at least 90% sequence identity to SEQ ID NO:24 and a light chain variable region having at least 90% sequence identity to SEQ ID NO:28. 
     
     
         88 . The method of  claim 85 , wherein the antibody comprises a heavy chain variable region comprising SEQ ID NO:24 and a light chain variable region comprising SEQ ID NO:28. 
     
     
         89 . The method of  claim 85 , wherein the antibody is a monoclonal antibody. 
     
     
         90 . The method of  claim 89 , wherein the antibody is a humanized antibody. 
     
     
         91 . The method of  claim 85 , further comprising administering to the subject at least one additional therapeutic agent. 
     
     
         92 . The method of  claim 91 , wherein at least one additional therapeutic agent is a chemotherapeutic agent. 
     
     
         93 . The method of  claim 92 , wherein the chemotherapeutic agent is a pyrimidine analog. 
     
     
         94 . The method of  claim 91 , wherein at least two additional therapeutic agents are chemotherapeutic agents. 
     
     
         95 . The method of  claim 94 , wherein one of the chemotherapeutic agents is a pyrimidine analog. 
     
     
         96 . The method of  claim 85 , wherein the colorectal cancer has metastasized. 
     
     
         97 . The method of  claim 95 , wherein the colorectal cancer has metastasized. 
     
     
         98 . The method of  claim 85 , wherein the antibody is administered after chemotherapy. 
     
     
         99 . The method of  claim 85 , wherein the antibody is administered after radiotherapy. 
     
     
         100 . A method of inhibiting the formation of a Notch1 intracellular domain (ICD), the method comprising: contacting a cell with an effective amount of an antibody that specifically binds human Notch1, wherein the antibody comprises:
 (a) a heavy chain CDR1 comprising RGYWIE (SEQ ID NO:15), a heavy chain CDR2 comprising QILPGTGRTNYNEKFKG (SEQ ID NO:16), and a heavy chain CDR3 comprising FDGNYGYYAMDY (SEQ ID NO:17); and   (b) a light chain CDR1 comprising RSSTGAVTTSNYAN (SEQ ID NO:18), a light chain CDR2 comprising GTNNRAP (SEQ ID NO:19), and a light chain CDR3 comprising ALWYSNHWVFGGGTKL (SEQ ID NO:20).   
     
     
         101 . The method of  claim 98 , wherein the antibody inhibits cleavage within the membrane proximal region of the extracellular domain of Notch1. 
     
     
         102 . An antibody that inhibits formation of a Notch1 intracellular domain (ICD), wherein the antibody comprises:
 (a) a heavy chain CDR1 comprising RGYWIE (SEQ ID NO:15), a heavy chain CDR2 comprising QILPGTGRTNYNEKFKG (SEQ ID NO:16), and a heavy chain CDR3 comprising FDGNYGYYAMDY (SEQ ID NO:17); and   (b) a light chain CDR1 comprising RSSTGAVTTSNYAN (SEQ ID NO:18), a light chain CDR2 comprising GTNNRAP (SEQ ID NO:19), and a light chain CDR3 comprising ALWYSNHWVFGGGTKL (SEQ ID NO:20).