Antibodies against epidermal growth factor receptor (egfr) and uses thereof

Abstract

Anti-EGFR antibodies, therapeutic compositions comprising combinations of anti-EGFR antibodies, as well as methods for using such antibodies and compositions to treat EGFR-related disorders (e.g., cancers), are disclosed.

What is claimed is: 
     
         1 . A monoclonal antibody preparation comprising at least one monoclonal antibody which binds EGFR extracellular domain and comprises heavy and light chain variable region sequences comprising CDR1, CDR2, and CDR3 sequences, wherein the at least one monoclonal antibody is selected from the group consisting of:
 antibody (a) having heavy chain CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 1, 2, and 3 respectively, and light chain CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 4, 5, and 6, respectively;   antibody (b) having heavy chain CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 7, 8, and 9, respectively, and light chain CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 10, 11 and 12, respectively; and   antibody (c) having heavy chain CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 13, 14, and 15, respectively, and light chain CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 16, 17, and 18, respectively.   
     
     
         2 . The monoclonal antibody preparation of  claim 1 , wherein the heavy and light chain variable region sequences of:
 antibody (a) comprise a heavy chain variable region comprising SEQ ID NO: 19 and a light chain variable region comprising SEQ ID NO: 20;   antibody (b) comprise a heavy chain variable region comprising SEQ ID NO: 21 and a light chain variable region comprising SEQ ID NO: 22; and   antibody (c) comprise a heavy chain variable region comprising SEQ ID NO: 23 and a light chain variable region comprising SEQ ID NO: 24.   
     
     
         3 . The monoclonal antibody preparation of  claim 1 , wherein each of (a), (b) and/or (c) binds to EGFR with a K D  of better than 100 nM. 
     
     
         4 . The monoclonal antibody preparation of  claim 3 , wherein each of (a), (b) and/or (c) binds to EGFR with a K D  of better than 10 nM. 
     
     
         5 . The monoclonal antibody preparation of  claim 3 , wherein each of (a), (b) and/or (c) binds to EGFR with a K D  of better than 1 nM. 
     
     
         6 . The monoclonal antibody preparation of  claim 1 , wherein each of (a), (b) and/or (c) is a human antibody. 
     
     
         7 . The monoclonal antibody preparation of  claim 1 , wherein each of (a), (b) and/or (c) is selected from the group consisting of a bispecific antibody, immunoconjugate, Fab, Fab′2, ScFv, avimer, nanobody and a domain antibody. 
     
     
         8 . The monoclonal antibody preparation of  claim 1 , wherein each of (a), (b) and/or (c) is selected from the group consisting of IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgAsec, IgD and IgE isotype antibodies. 
     
     
         9 . A pharmaceutical composition comprising the monoclonal antibody preparation of  claim 6  and a pharmaceutically acceptable carrier. 
     
     
         10 . A method of treating cancer in a subject, comprising administering to the subject an effective amount of the pharmaceutical composition of  claim 9 . 
     
     
         11 . A composition comprising all three of monoclonal antibodies (a), (b), and (c) of  claim 1  in a pharmaceutically acceptable carrier. 
     
     
         12 . The composition of  claim 11 , wherein:
 antibody (a) comprises a heavy chain variable region comprising SEQ ID NO: 19 and a light chain variable region comprising SEQ ID NO: 20;   antibody (b) comprises a heavy chain variable region comprising SEQ ID NO: 21 and a light chain variable region comprising SEQ ID NO: 22; and   antibody (c) comprised a heavy chain variable region comprising SEQ ID NO: 23 and a light chain variable region comprising SEQ ID NO: 24.   
     
     
         13 . The composition of  claim 12 , wherein each of monoclonal antibodies (a), (b) and (c) binds to EGFR with a K D  of better than 100 nM. 
     
     
         14 . The composition of  claim 13 , wherein each of monoclonal antibodies (a), (b) and (c) binds to EGFR with a K D  of better than 10 nM. 
     
     
         15 . The composition of  claim 13 , wherein each of monoclonal antibodies (a), (b) and (c) binds to EGFR with a K D  of better than 1 nM. 
     
     
         16 . The composition of  claim 11 , wherein each of monoclonal antibodies (a), (b) and (c) is a human antibody. 
     
     
         17 . The composition of  claim 11 , wherein one or more of monoclonal antibodies (a), (b), and (c) is independently selected from the group consisting of a bispecific antibody, immunoconjugate, Fab, Fab′2, ScFv, avimer, nanobody and a domain antibody. 
     
     
         18 . The composition of  claim 11 , wherein each of monoclonal antibodies (a), (b), and (c) is independently selected from the group consisting of IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgAsec, IgD and IgE isotype antibodies. 
     
     
         19 . A method of treating cancer in a subject, comprising administering to the subject an effective amount of the composition of  claim 16 . 
     
     
         20 . The composition of  claim 11 , wherein antibodies (a), (b), and (c) are present at a molar ratio of 2:2:1 to each other. 
     
     
         21 . The composition of  claim 20 , wherein each of antibody (a), antibody (b) and antibody (c) binds to EGFR with a K D  of better than 100 nM. 
     
     
         22 . The composition of  claim 21 , wherein each of antibody (a), antibody (b) and antibody (c) binds to EGFR with a K D  of better than 10 nM. 
     
     
         23 . The composition of  claim 20 , wherein antibody (a) binds to EGFR with a K D  in a range of 1×10 −9  M to 1.1×10 −11  M, antibody (b) binds to EGFR with a K D  in a range of 1×10 −9  M to 7.0×10 −11  M and antibody (c) binds to EGFR with a K D  in a range of 1×10 −9  M to 3.6×10 −10  M. 
     
     
         24 . The composition of  claim 20 , wherein each of antibody (a), antibody (b) and antibody (c) are human antibodies. 
     
     
         25 . A kit comprising the composition of  claim 20  in a container. 
     
     
         26 . A method of treating cancer in a subject, comprising administering to the subject an effective amount of the composition of  claim 20 . 
     
     
         27 . A method of preparing an anti-EGFR antibody composition, the method comprising combining in a single composition:
 all three of monoclonal antibodies (a), (b), and (c) of  claim 1 , and a pharmaceutically acceptable carrier;   wherein (a), (b) and (c) are combined at a molar ratio of 2:2:1 to each other.   
     
     
         28 . The method of  claim 27  wherein:
 antibody (a) comprises a heavy chain variable region comprising SEQ ID NO: 19 and a light chain variable region comprising SEQ ID NO:20; 
 antibody (b) comprises a heavy chain variable region comprising SEQ ID NO: 21 and a light chain variable region comprising SEQ ID NO: 22; and 
 antibody (c) comprises a heavy chain variable region comprising SEQ ID NO: 23 and a light chain variable region comprising SEQ ID NO: 24. 
 
     
     
         29 . A method of treating a subject with anti-EGFR antibodies, the method comprising administering to the subject:
 (a) a monoclonal antibody comprising heavy chain CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 1, 2, and 3 respectively, and light chain CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 4, 5, and 6, respectively;   (b) a monoclonal antibody comprising heavy chain CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 7, 8, and 9, respectively, and light chain CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 10, 11 and 12, respectively; and   (c) a monoclonal antibody comprising heavy chain CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 13, 14, and 15 respectively, and light chain CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 16, 17, and 18, respectively;   wherein (a), (b) and (c) are administered to the subject at a molar ratio of 2:2:1 to each other.   
     
     
         30 . The method of  claim 29  wherein
 antibody (a) comprises a heavy chain variable region comprising SEQ ID NO: 19 and a light chain variable region comprising SEQ ID NO:20; 
 antibody (b comprises a heavy chain variable region comprising SEQ ID NO: 21 and a light chain variable region comprising SEQ ID NO: 22; and 
 antibody (c) comprises a heavy chain variable region comprising SEQ ID NO: 23 and a light chain variable region comprising SEQ ID NO: 24.