Monoclonal antibodies against her2 epitope and methods of use thereof

Abstract

This invention provides fully human monoclonal antibodies that recognize HER2. The invention further provides methods of using such monoclonal antibodies in a variety of therapeutic, diagnostic, and prophylactic indications.

What is claimed is: 
     
         1 . An isolated antibody or antigen binding fragment thereof that specifically binds to an epitope of the human HER2 receptor, wherein the isolated antibody or antigen binding fragment thereof binds at least a portion of the N-terminus of domain IV of human HER2 receptor but does not cross-compete with an antibody that binds to epitope 4D5 of the human HER2 receptor, and wherein the antibody or antigen binding fragment thereof specifically binds to an epitope of the human HER2 receptor that comprises amino acid residues 452 to 531 of the extracellular domain of the human HER2 receptor. 
     
     
         2 . The isolated antibody or antigen binding fragment thereof of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof specifically binds to an epitope of the human HER2 receptor that comprises amino acid residues 520 to 531 of the extracellular domain of the human HER2 receptor. 
     
     
         3 . The isolated antibody or antigen binding fragment thereof of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof specifically binds to an epitope of the human HER2 receptor that comprises amino acid residues E521, L525 and R530 of the extracellular domain of the human HER2 receptor. 
     
     
         4 . The isolated antibody or antigen binding fragment thereof of  claim 1 , wherein the antibody or antigen binding fragment thereof competes for binding the human HER2 receptor with an antibody or antigen binding fragment thereof that comprises a variable heavy chain complementarity determining region 1 (CDRH1) comprising the amino acid sequence FTFSSYSMN (SEQ ID NO: 25); a variable heavy chain complementarity determining region 2 (CDRH2) comprising the amino acid sequence YISSSSSTIYYADSVKG (SEQ ID NO: 26); a variable heavy chain complementarity determining region 3 (CDRH3) comprising the amino acid sequence GGHGYFDL (SEQ ID NO: 27); a variable light chain complementarity determining region 1 (CDRL1) comprising the amino acid sequence RASQSVSSSYLA (SEQ ID NO: 28); a variable light chain complementarity determining region 2 (CDRL2) comprising the amino acid sequence GASSRAT (SEQ ID NO: 21); and a variable light chain complementarity determining region 3 (CDRL3) comprising the amino acid sequence QQYHHSPLT (SEQ ID NO: 29). 
     
     
         5 . The isolated antibody or antigen binding fragment thereof of  claim 1 , wherein the antibody or antigen binding fragment thereof comprises a variable heavy chain complementarity determining region 1 (CDRH1) comprising the amino acid sequence FTFSSYSMN (SEQ ID NO: 25) or an amino acid sequence at least 80% identical thereto; a variable heavy chain complementarity determining region 2 (CDRH2) comprising the amino acid sequence YISSSSSTIYYADSVKG (SEQ ID NO: 26) or an amino acid sequence at least 80% identical thereto; a variable heavy chain complementarity determining region 3 (CDRH3) comprising the amino acid sequence GGHGYFDL (SEQ ID NO: 27) or an amino acid sequence at least 80% identical thereto; a variable light chain complementarity determining region 1 (CDRL1) comprising the amino acid sequence RASQSVSSSYLA (SEQ ID NO: 28) or an amino acid sequence at least 80% identical thereto; a variable light chain complementarity determining region 2 (CDRL2) comprising the amino acid sequence GASSRAT (SEQ ID NO: 21) or an amino acid sequence at least 80% identical thereto; and a variable light chain complementarity determining region 3 (CDRL3) comprising the amino acid sequence QQYHHSPLT (SEQ ID NO: 29) or an amino acid sequence at least 80% identical thereto, and wherein the antibody or antigen binding fragment thereof exhibits a K d  less than 100 nM for the human HER2 receptor. 
     
     
         6 . The isolated antibody or antigen binding fragment thereof of  claim 5 , wherein any amino acid changes relative to SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 21, or SEQ ID NO: 29 are conservative amino acid substitutions. 
     
     
         7 . The isolated antibody or antigen binding fragment thereof of  claim 1 , wherein the antibody or antigen binding fragment thereof comprises a variable heavy chain comprising a CDRH1, CDRH2, and CDRH3 and comprising the amino acid sequence of SEQ ID NO: 13, or an amino acid sequence at least 80% identical thereto and a variable light chain comprising a CDRL1, CDRL2, and CDRL3 and comprising the amino acid sequence of SEQ ID NO: 14, or an amino acid sequence at least 80% identical thereto, and wherein the antibody or antigen binding fragment thereof exhibits a K d  less than 100 nM for the human HER2 receptor. 
     
     
         8 . The isolated antibody or antigen binding fragment thereof of  claim 7 , wherein any amino acid changes relative to CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, CDRL3, given by SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 21, or SEQ ID NO: 29, respectively, are conservative amino acid substitutions. 
     
     
         9 . The isolated antibody or antigen binding fragment thereof of  claim 1 , wherein the antibody or antigen binding fragment thereof is a monoclonal antibody, a domain antibody, a single chain antibody, a Fab fragment, a F(ab′) 2  fragment, a single chain variable fragment (scFv), a scFv-Fc fragment, a single chain antibody (scAb), a domain antibody (dAb), a single domain heavy chain antibody, or a single domain light chain antibody. 
     
     
         10 . The isolated antibody or antigen binding fragment thereof of  claim 1 , wherein the antibody or antigen binding fragment thereof is a rabbit, mouse, chimeric, humanized or fully human monoclonal antibody. 
     
     
         11 . The isolated antibody or antigen binding fragment thereof of  claim 1 , wherein the antibody or antigen binding fragment thereof is an IgG isotype. 
     
     
         12 . The isolated antibody or antigen binding fragment thereof of  claim 1 , wherein the antibody or antigen binding fragment thereof is an IgG 1  isotype. 
     
     
         13 . An isolated nucleic acid molecule encoding the isolated antibody or antigen binding fragment thereof of  claim 1 . 
     
     
         14 . A vector comprising the isolated nucleic acid molecule of  claim 13 . 
     
     
         15 . A method of producing an isolated antibody or antigen binding fragment thereof by culturing a cell under conditions that lead to expression of the isolated antibody or antigen binding fragment thereof, wherein the cell comprises the vector of  claim 14 . 
     
     
         16 . A conjugate comprising the isolated antibody or antigen binding fragment of  claim 1  and one or more therapeutic or diagnostic agents (D), each of which is independently connected directly or indirectly to the antibody or antigen binding fragment thereof. 
     
     
         17 . The conjugate of  claim 16 , further comprising one or more polymeric scaffolds connected both to the antibody or antigen binding fragment thereof, wherein each of the one or more D is independently connected to the antibody or antigen binding fragment thereof via the one or more polymeric scaffolds. 
     
     
         18 . The conjugate of  claim 17 , wherein each of the one or more polymeric scaffolds independently comprises poly(1-hydroxymethylethylene hydroxymethyl-formal) (PHF) having a molecular weight ranging from about 2 kDa to about 40 kDa. 
     
     
         19 . The conjugate of  claim 18 , wherein each of the one or more polymeric scaffolds independently is of Formula (Ic): 
       
         
           
           
               
               
           
         
         wherein: 
         L D1  is a carbonyl-containing moiety; 
         each occurrence of 
       
       
         
           
           
               
               
           
         
          in is independently a first linker that contains a biodegradable bond so that when the bond is broken, D is released in an active form for its intended therapeutic effect; and the 
       
       
         
           
           
               
               
           
         
          between L D1  and D denotes direct or indirect attachment of D to L D1 ; 
         each occurrence of 
       
       
         
           
           
               
               
           
         
          is independently a second linker not yet connected to the isolated antibody or antigen binding fragment thereof, in which L P2  is a moiety containing a functional group that is yet to form a covalent bond with a functional group of the isolated antibody or antigen binding fragment thereof, and the 
       
       
         
           
           
               
               
           
         
          between L D1  and L P2  denotes direct or indirect attachment of L P2  to L D1 , and each occurrence of the second linker is distinct from each occurrence of the first linker; 
         each occurrence of 
       
       
         
           
           
               
               
           
         
          is independently a third linker that connects each D-carrying polymeric scaffold to the isolated antibody or antigen binding fragment thereof, in which the terminal 
       
       
         
           
           
               
               
           
         
          attached to L P2  denotes direct or indirect attachment of L P2  to the isolated antibody or antigen binding fragment thereof upon formation of a covalent bond between a functional group of L P2  and a functional group of the isolated antibody or antigen binding fragment thereof; and each occurrence of the third linker is distinct from each occurrence of the first linker; 
         m is an integer from 1 to about 300, 
         m 1  is an integer from 1 to about 140, 
         m 2  is an integer from 1 to about 40, 
         m 3  is an integer from 0 to about 18, 
         m 4  is an integer from 1 to about 10; 
         the sum of m, m 1 , m 2 , m 3 , and m 4  ranges from about 15 to about 300; and 
         the total number of L P2  connected to the isolated antibody or antigen binding fragment thereof is 10 or less. 
       
       
         
           
           
               
               
           
         
       
     
     
         20 . The conjugate of  claim 19 , wherein each occurrence of 
       
         
           
           
               
               
           
         
       
       is independently —C(═O)—X—(CH 2 ) v —C(═O)—NH—(CH 2 ) u —NHC(═O)—(CH 2 ) w —(OCH 2 ) x —NHC(═O)—(CH 2 ) y -M, in which X is CH 2 , O, or NH, each of v, u, w, x and y independently is an integer from 1 to 6, and M is 
       
         
           
           
               
               
           
         
       
       wherein one of X a  and X b  is H and the other is a water-soluble maleimido blocking moiety, or X a  and X b , together with the carbon atoms to which they are attached form a carbon-carbon double bond. 
     
     
         21 . The conjugate of  claim 20 , wherein each of v, u, w, x and y is 2. 
     
     
         22 . The conjugate of  claim 21 , wherein each of the one or more polymeric scaffolds independently is of Formula (Id): 
       
         
           
           
               
               
           
         
       
       wherein
 m 3a  is an integer from 0 to about 17, 
 m 3b  is an integer from 1 to about 8, and 
 the terminal 
 
       
         
           
           
               
               
           
         
          denotes the direct attachment of the one or more polymeric scaffolds to the isolated antibody or antigen binding fragment thereof. 
       
     
     
         23 . A method of alleviating a symptom of a cancer in a human subject in need thereof, the method comprising administering a conjugate according to  claim 16  to the subject in an amount sufficient to alleviate the symptom of the cancer, wherein the cancer is selected from the group consisting of anal cancer, astrocytoma, leukemia, lymphoma, head and neck cancer, liver cancer, testicular cancer, cervical cancer, sarcoma, hemangioma, esophageal cancer, eye cancer, laryngeal cancer, mouth cancer, mesothelioma, skin cancer, myeloma, oral cancer, rectal cancer, throat cancer, bladder cancer, breast cancer, uterine cancer, ovarian cancer, prostate cancer, lung cancer, non-small cell lung cancer (NSCLC), colon cancer, pancreatic cancer, renal cancer, and gastric cancer. 
     
     
         24 . The method of  claim 23 , wherein the cancer is selected from the group consisting of breast cancer, gastric cancer, non-small cell lung cancer (NSCLC), and ovarian cancer.