Monoclonal antibodies to human fibroblast growth factor receptor 2 (hfgfr2) and methods of use thereof

Abstract

The present invention provides a pharmaceutical composition or a diagnostic composition targeting human fibroblast growth factor receptor 2 (hFGFR2).

1 - 47 . (canceled) 
     
     
         48 . A monoclonal antibody or an antigen binding fragment thereof, wherein the monoclonal antibody consists of
 a heavy chain comprising a complementarity determining region H1 (CDRH1) consisting of the amino acid sequence represented by SEQ ID NO: 24, a CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 25, and a CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 26; and   a light chain comprising a CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 27, a CDRL2 consisting of the amino acid sequence represented by SEQ ID NO:   28, and a CDRL3 consisting of the amino acid sequence represented by SEQ ID NO: 29.   
     
     
         49 . The monoclonal antibody or antigen binding fragment thereof according to  claim 48 , wherein the monoclonal antibody or antigen binding fragment thereof specifically binds to non-denatured human fibroblast growth factor receptor 2 (hFGFR2) IIIc and denatured hFGFR2 IIIc in a preparation fixed in formalin. 
     
     
         50 . The monoclonal antibody or antigen binding fragment thereof according to  claim 48 , wherein the monoclonal antibody comprises a heavy chain variable region and a light chain variable region described in any one of the following (i) to (iv):
 (i) a heavy chain variable region of SEQ ID NO: 3 and a light chain variable region of SEQ ID NO: 6;   (ii) a heavy chain variable region in which the framework region of the heavy chain variable region is 95% of more identical to the framework region of SEQ ID NO: 3 and a light chain variable region in which the framework region of the light chain variable region is 95% of more identical to the framework region of SEQ ID NO: 6;   (iii) a heavy chain variable region derived from SEQ ID NO: 3 by incorporating a substitution, deletion, insertion, or addition of 1 or more amino acids into the framework region, and a light chain variable region derived from SEQ ID NO: 6 by incorporating a substitution, deletion, insertion, or addition of 1 or more amino acids into the framework region; and   (iv) a heavy chain variable region encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide that encodes SEQ ID NO: 3, and a light chain variable region encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide that encodes SEQ ID NO: 6.   
     
     
         51 . The monoclonal antibody or antigen binding fragment thereof which has the following properties (i) to (iii):
 (i) specifically binds to non-denatured human fibroblast growth factor receptor 2 (hFGFR2) IIIc;   (ii) specifically binds to none of the following: non-denatured human fibroblast growth factor receptor 1 (hFGFR1); non-denatured human fibroblast growth factor receptor 3 (hFGFR3); and non-denatured human fibroblast growth factor receptor 4 (hFGFR4); and   (iii) specifically binds to denatured hFGFR2 IIIc in a preparation fixed in formalin,   wherein the monoclonal antibody or antigen binding fragment thereof binds to a site on hFGFR2 Mc which is recognized by an antibody or an antigen binding fragment thereof according to claim  3  or competes with an antibody or an antigen binding fragment thereof according to claim  3  for binding to hFGFR2 IIIc.   
     
     
         52 . The monoclonal antibody or antigen binding fragment thereof according to  claim 48 , wherein the monoclonal antibody comprises the amino acid sequence of a heavy chain represented by SEQ ID NO: 15 and the amino acid sequence of a light chain represented by SEQ ID NO: 19. 
     
     
         53 . The monoclonal antibody or antigen binding fragment thereof according to  claim 48 , wherein the monoclonal antibody or antigen binding fragment thereof binds to neither non-denatured human fibroblast growth factor receptor 2 (hFGFR2) IIIb nor denatured hFGFR2 IIIb in a preparation fixed in formalin. 
     
     
         54 . A monoclonal antibody or an antigen binding fragment thereof which is obtained by a method comprising the following steps (i) and (ii):
 (i) culturing a cell comprising a polynucleotide encoding a monoclonal antibody according to  claim 48  or a vector comprising a polynucleotide encoding a monoclonal antibody according to  claim 48 ; and   (ii) recovering the monoclonal antibody or antigen binding fragment thereof from the cultures of step (i).   
     
     
         55 . A composition comprising a monoclonal antibody or an antigen binding fragment thereof according to  claim 48 . 
     
     
         56 . A composition comprising a monoclonal antibody or an antigen binding fragment thereof according to  claim 48 , wherein the composition can detect or assay for hFGFR2 IIIc in a tissue preparation which is paraffin-embedded and then deparaffinized, the tissue preparation comprising the monoclonal antibody or antigen binding fragment thereof according to claim  1 . 
     
     
         57 . The composition according to  claim 56 , wherein the preparation is subjected to enzymatic treatment following the deparaffinization treatment. 
     
     
         58 . The composition according to  claim 57 , wherein the enzymatic treatment is the reaction of protease at 20 to 38° C. 
     
     
         59 . A pharmaceutical composition comprising an antibody or an antigen binding fragment thereof according to  claim 48  that specifically binds to hFGFR2 Mc. 
     
     
         60 . A method for detecting or assaying hFGFR2 IIIc, comprising the following step (i) or steps (i) and (ii):
 (i) contacting a monoclonal antibody or an antigen binding fragment of the antibody according to  claim 48  or a composition comprising a monoclonal antibody or an antigen binding fragment thereof according to  claim 48  with a test preparation; and   (ii) determining the test preparation to be positive when hFGFR2 IIIc is detected or assayed in the test preparation or when the expression level of hFGFR2 IIIc in the test preparation is equivalent to or higher than predetermined references; and determining the test preparation to be negative when hFGFR2 Mc is not detected or assayed in the test preparation or when the expression level of hFGFR2 IIIc in the test preparation is equivalent to or lower than the predetermined references.   
     
     
         61 . The method according to  claim 60 , wherein the method is used in a method for testing or diagnosing a hFGFR2 IIIc-positive patient. 
     
     
         62 . The method according to  claim 61 , wherein the testing or diagnosing is for cancer. 
     
     
         63 . A method for identifying a suitable individual for treating with a pharmaceutical composition comprising an antibody specifically binding to hFGFR2 or an antigen binding fragment of the antibody, the method comprising the following step (i) or steps (i) and (ii) or steps (i)-(iii):
 (i) contacting an antibody or an antigen binding fragment of the antibody according to  claim 48  or a composition comprising a monoclonal antibody or an antigen binding fragment thereof according to  claim 48  with an individual-derived sample;   (ii) determining the individual to be positive when hFGFR2 IIIc is detected or assayed in the individual-derived sample or when the expression level of hFGFR2 IIIn the individual derived sample is equivalent to or higher than predetermined references, and determining the individual to be negative when hFGFR2 IIIc is not detected or assayed in the individual-derived sample or when the expression level of hFGFR2 IIIc in the individual-derived sample is equivalent to or lower than the predetermined references; and   (iii) treating the individual who is determined to be positive with the pharmaceutical composition.   
     
     
         64 . A method for treating a patient that is positive for hFGFR2 IIIc, comprising administering a pharmaceutical composition comprising an antibody or antigen binding fragment thereof according to  claim 48  to a patient in need thereof. 
     
     
         65 . A kit for testing or diagnosing a hFGFR2 IIIc-patient disease, comprising an antibody or an antigen binding fragment of the antibody according to  claim 48 . 
     
     
         66 . The kit according to  claim 65 , wherein the testing or diagnosing is for cancer.