Chimeric Anti-VEGF-D Antibodies and Humanized Anti-VEGF-D Antibodies and Methods of Using the Same

Abstract

The present invention relates to materials and methods for modulating angiogenesis and lymphangiogenesis. The compositions of the invention provide chimeric and/or humanized VEGF-D antibody substances, antibodies, polypeptides and fragments thereof useful for modulating angiogenesis and lymphangiogenesis in a subject.

1 . A chimeric or humanized antibody or antibody fragment comprising:
 (a) an antibody that specifically binds Vascular Endothelial Growth Factor-D (VEGF-D), comprising
 i) a light chain variable region comprising complementarity determining regions (L-CDR), wherein said L-CDR comprise amino acid sequences identical to SEQ ID NOs: 47-49, or comprise 1 or 2 amino acid substitutions in the L-CDR compared to a light chain variable region comprising SEQ ID NOs: 47-49; 
 ii) a heavy chain variable region comprising complementarity determining regions (H-CDR), wherein said H-CDR comprise amino acid sequences identical to SEQ ID NOs: 50-52, or comprise 1 or 2 amino acid substitutions in the H-CDR compared to a heavy chain variable region comprising SEQ ID NOs: 50-52; 
 iii) a human antibody light chain constant region; and 
 iv) a human antibody heavy chain constant region; or 
   (b) a fragment of (a) that binds VEGF-D.   
     
     
         2 - 3 . (canceled) 
     
     
         4 . A chimeric or humanized antibody or antibody fragment comprising:
 (a) an antibody that specifically binds Vascular Endothelial Growth Factor-D (VEGF-D), comprising
 i) heavy chain complementarity determining regions from a mouse antibody and framework regions from non-murine source, wherein the heavy chain variable region (VH) comprises complementarity determining regions (CDR) at least 90% identical to the amino acid sequences: 
 H-CDR1 set out in SEQ ID NO: 50 
 H-CDR2 set out in SEQ ID NO: 51 
 H-CDR3 set out in SEQ ID NO: 52 
 ii) light chain complementarity determining regions from a mouse antibody and framework regions from non-murine source, wherein the light chain variable region (VL) comprises complementarity determining regions (CDR) at least 90% identical to the amino acid sequences: 
 L-CDR1: set out in SEQ ID NO: 47 
 L-CDR2: set out in SEQ ID NO: 48 
 L-CDR3: set out in SEQ ID NO: 49; 
 iii) a human antibody light chain constant region; and 
 iv) a human antibody heavy chain constant region; or 
   (b) a fragment of (a) that binds VEGF-D.   
     
     
         5 . The antibody or antibody fragment of  claim 1 , wherein the framework regions of the heavy chain variable region and the framework regions of the light chain variable region comprise human framework regions. 
     
     
         6 - 7 . (canceled) 
     
     
         8 . The antibody or antibody fragment according to  claim 1 , wherein the light chain constant region is a kappa or lambda light chain. 
     
     
         9 . The antibody or antibody fragment according to  claim 1 , wherein the heavy chain constant region is selected from the group consisting of a constant region from an IgM chain, an IgG chain, an IgA chain, an IgE chain, an IgD chain, fragments thereof, and combinations thereof. 
     
     
         10 . The antibody or antibody fragment according to  claim 1 , wherein the heavy chain constant region comprises an IgG chain selected from the group consisting of IgG1, IgG2, IgG3, IgG4, fragments thereof, and combinations thereof. 
     
     
         11 . The antibody or antibody fragment of  claim 10  wherein the constant region comprises at least one of CH1, CH2, and CH3 regions of a human IgG1 heavy chain constant region. 
     
     
         12 . The antibody substance or antibody fragment according to  claim 1  that comprises a Fab fragment of the antibody. 
     
     
         13 . The antibody substance or antibody fragment according to  claim 1  that is a monoclonal antibody. 
     
     
         14 - 32 . (canceled) 
     
     
         33 . The antibody or antibody fragment according to  claim 1  that inhibits VEGF-D binding to VEGFR-2 or to VEGFR-3. 
     
     
         34 . (canceled) 
     
     
         35 . The antibody, or fragment according to  claim 1  that inhibits VEGF-D stimulation of endothelial cell growth. 
     
     
         36 . A composition comprising the antibody or fragment according to  claim 1  and in a pharmaceutically acceptable carrier. 
     
     
         37 - 50 . (canceled) 
     
     
         51 . An isolated polynucleotide comprising a nucleotide sequence that encodes the antibody or fragment according to  claim 1 , or encodes a light chain polypeptide or a heavy chain polypeptide of said antibody. 
     
     
         52 - 59 . (canceled) 
     
     
         60 . A method for inhibiting VEGF-D mediated cell growth, migration, or differentiation, comprising administering to a human subject an antibody or fragment according to  claim 1 , in an amount effective to inhibit VEGF-D interaction with VEGFR-2 or VEGFR-3. 
     
     
         61 . The method according to  claim 60 , wherein the antibody or fragment is administered in an amount effective to inhibit angiogenesis or lymphangiogenesis in the human subject. 
     
     
         62 . The method of  claim 61  wherein the subject is suffering from a condition or disorder resulting from aberrant angiogenesis or lymphangiogenesis. 
     
     
         63 . The method of  claim 62  wherein the condition or disorder is cancer. 
     
     
         64 . The method of  claim 63  wherein the antibody or fragment is administered in combination with a second agent selected from the group consisting of a chemotherapeutic agent, a radiotherapeutic agent, or radiation therapy. 
     
     
         65 . (canceled) 
     
     
         66 . The chimeric or humanized antibody or antibody fragment of  claim 4  wherein the light chain CDRs comprise amino acid sequences: SEQ ID NO: 47, SEQ ID NO: 48 and SEQ ID NO: 49. 
     
     
         67 . The antibody or antibody fragment of  claim 4 , wherein the framework regions of the heavy chain variable region and the framework regions of the light chain variable region comprise human framework regions.