Anti-cd147 antibodies, methods and uses

Abstract

The present invention provides antibodies immunospecific for human CD147 capable of blocking bioactivity of CD147 associated with malignant disease such as the stimulation of MMPs from fibroblast cells by tumor cells, the release of VEGF, and the promotion of angiogenesis. The antibodies of the present invention of are useful in treating malignant disease and those diseases in which CD147 activity is plays a pathogenic role, such as diseases of the eye, lung, and cardiovascular system.

What is claimed: 
     
         1 . An isolated monoclonal antibody or antigen-binding fragment thereof that competes for binding to the epitope on CD147 bound by the monoclonal antibody selected from the group consisting of 2H3, 4A5, and 5F6 having the amino acid sequences of the light chain complementarity determining regions (CDRs) of one of SEQ ID NOs: 9, 11, and 13 and the amino acid sequences of the heavy chain CDRs of one of SEQ ID NOs: 10, 12, and 14. 
     
     
         2 . An isolated antibody having heavy chain CDR1, CDR 2 and CDR3 (Hc-CDR1, Hc-CDR2 and Hc-CDR3) amino acid sequences selected from the sequences shown in SEQ ID NOs: 10, 12 and 14 respectively and a light chain CDR3 (Lc-CDR3) as shown in Formula (I):
   Gln Gln Xaa 1  Tyr Ser Xaa 2  Pro Xaa 3 Thr  (I)
   wherein Xaa 1  is Tyr or Asp; Xaa 2  is Tyr or Ser; Xaa 3  is Phe or Tyr or absent; and Xaa 4  is Thr or Phe; and light chain CDR1 (Lc-CDR1) and light chain CDR2 (Lc-CDR2) amino acid sequences selected from the sequences as shown in SEQ ID NOs: 9 and 11, respectively.   
     
     
         3 . An isolated antibody having Hc-CDR1, Hc-CDR2 and Hc-CDR3 amino acid sequences shown in SEQ ID NOs: 10 and Lc-CDR1 Lc-CDR2, and Lc-CDR3 amino acid sequences as shown in SEQ ID NOs: 9. 
     
     
         4 . An isolated antibody having Hc-CDR1, Hc-CDR2 and Hc-CDR3 amino acid sequences shown in SEQ ID NOs: 12 and Lc-CDR1 Lc-CDR2, and Lc-CDR3 amino acid sequences as shown in SEQ ID NOs: 11. 
     
     
         5 . An isolated antibody having Hc-CDR1, Hc-CDR2 and Hc-CDR3 amino acid sequences shown in SEQ ID NOs: 14 and Lc-CDR1 Lc-CDR2, and Lc-CDR3 amino acid sequences as shown in SEQ ID NOs: 13. 
     
     
         6 . An isolated antibody having Hc-CDR1, Hc-CDR2 and Hc-CDR3 amino acid sequences shown in SEQ ID NOs: 16 and Lc-CDR1 Lc-CDR2, and Lc-CDR3 amino acid sequences as shown in SEQ ID NOs: 15. 
     
     
         7 . An isolated recombinant anti-CD147 antibody or antigen-binding fragment thereof, said antibody comprising a human constant region, wherein said antibody or antigen-binding fragment (i) has epitopic specificity identical to the 4A5 Mab as determined by H/D exchange on CD147, comprising SEQ ID NO: 11 and 12, and (ii) binds to the epitope of human CD 147 with a K D  of at least 1×10 −7  M. 
     
     
         8 . A humanized antibody comprising a humanized heavy chain and humanized light chain, wherein:
 a) the humanized heavy chain variable region comprises three complementarity determining regions (CDRS) from the mouse 4A5 heavy chain (SEQ ID NO: 12) and a framework from a human acceptor antibody heavy chain, and   b) the humanized light chain variable region comprises three complementarity determining regions from the mouse 4A5 light chain (SEQ ID NO: 11) and a framework from a human acceptor antibody light chain; and   c) wherein the humanized antibody specifically binds to a CD147 antigen on the surface of MDA-MB-231 cells.   
     
     
         9 . The antibody or antigen-binding fragment of any of  claims 1 - 8 , wherein said binding of the antibody or antigen-binding fragment to human CD147 inhibits a pathological activity of human CD147. 
     
     
         10 . A method of inhibiting tumor growth, comprising contacting a tumor with an effective amount of an isolated monoclonal antibody or antigen binding portion thereof that binds to an epitope comprising amino acids 64-75 of SEQ ID NO: 1 and inhibits the production of MMP-2 from human fibroblasts cells in the presence of human MDA-MB-231 cells. 
     
     
         11 . The method of  claim 10 , wherein the tumor is selected from the group consisting of colon carcinoma, breast tumor, prostate tumor, squamous cell carcinoma, and lung cancer. 
     
     
         12 . The method of  claim 10 , wherein the isolated monoclonal antibody or antigen binding portion thereof is administered systemically. 
     
     
         13 . The method of  claim 10 , wherein the isolated monoclonal antibody is administered site specifically. 
     
     
         14 . A method of inhibiting angiogenesis, comprising contacting a tissue with an effective amount of an isolated monoclonal antibody or antigen binding portion thereof that binds to an epitope comprising amino acids 64-75 of SEQ ID NO: 1 (DALPGQKTEF) and inhibits the production of MMP-2 from human fibroblasts cells in the presence of human MDA-MB-231 cells. 
     
     
         15 . A method of immunizing a subject predisposed to a condition involving pathologic CD147 bioactivity comprising administering a polypeptide comprising residues 64-75 of SEQ ID NO: 1 (DALPGQKTEF). 
     
     
         16 . An article of manufacture comprising a pharmaceutically acceptable formulation comprising the antibody of any of  claims 1 - 8 .