Combinatorial therapy

Abstract

The present invention relates to the use of VEGF antagonists and alpha5beta1 antagonists for treating cancer and inhibiting angiogenesis and/or vascular permeability, including inhibiting abnormal angiogenesis in diseases. The present invention also relates to use of a VEGFR agonists and alpha5beta1 agonists to promote angiogenesis and vascular permeability. The present invention also relates to new anti-alpha5beta1 antibodies, compositions and kits comprising them and methods of making and using them.

1 . An antibody that can bind human alpha5beta1 and competitively inhibit the binding of an anti-alpha5beta1 antibody to human alpha5beta1, wherein the anti-alpha5beta1 antibody is produced by a hybridoma selected from the group consisting of the hybridoma deposited as Alpha5/beta1 7H5.4.2.8 (ATCC No. PTA-7421) and the hybridoma deposited as Alpha5/beta1 7H12.5.1.4 (ATCC No. PTA-7420) in the ATCC on Mar. 7, 2006. 
     
     
         2 . (canceled) 
     
     
         3 . An antibody comprising the variable heavy (VH) and variable light (VL) domain sequence of the antibody produced by the hybridoma deposited as Alpha5/beta1 7H5.4.2.8 (ATCC No. PTA-7421) in the ATCC on Mar. 7, 2006. 
     
     
         4 . An antibody comprising the variable heavy (VH) and variable light (VL) domain sequence of the antibody produced by the hybridoma deposited as Alpha5/beta1 7H12.5.1.4 (ATCC No. PTA-7420) in the ATCC on Mar. 7, 2006. 
     
     
         5 . The antibody according to  claim 3  or  4 , wherein the antibody is a humanized or chimeric antibody. 
     
     
         6 . The antibody according to  claim 1 , wherein the antibody binds a human alpha5beta1 or alpha5 with a Kd between 500 nM and 1 pM. 
     
     
         7 . (canceled) 
     
     
         8 . The antibody according to  claim 3  or  4 , wherein the antibody comprises a Fc sequence of a human IgG. 
     
     
         9 . The antibody according to  claim 8 , wherein the human IgG is IgG10 IgG4. 
     
     
         10 . The antibody according to  claim 8 , wherein the antibody comprises a Fc sequence that lacks antibody dependent cellular cytotoxicity (ADCC) effector function. 
     
     
         11 . The antibody according to  claim 3  or  4 , wherein the antibody is selected from the group consisting of a Fab, Fab′, a F(ab)′2, single-chain Fv (scFv), an Fv fragment; a diabody and a linear antibody. 
     
     
         12 . The antibody according to  claim 3  or  4 , wherein the antibody is a multi-specific antibody. 
     
     
         13 . The antibody according to any one of  claims 1 ,  3  or  4  conjugated to a therapeutic agent. 
     
     
         14 . The antibody according to  claim 13 , wherein the therapeutic agent is selected from the group consisting of a cytotoxic agent, a radioisotope and a chemotherapeutic agent. 
     
     
         15 . The antibody according to any one of  claims 1 ,  3  or  4  conjugated to a label. 
     
     
         16 . The antibody according to  claim 15 , wherein the label is selected from the group consisting of a radioisotope, fluorescent dye and enzyme. 
     
     
         17 . A isolated nucleic acid molecule that encodes the variable heavy chain domain (VH) or the variable light chain domain (VL) or both V H  and V L  domains of any one of the antibodies of  claims 1 ,  3  or  4 . 
     
     
         18 . An expression vector encoding the nucleic acid molecule of  claim 17 . 
     
     
         19 . A cell comprising the nucleic acid molecule of  claim 17 . 
     
     
         20 . The cell according to  claim 19  wherein the cell is the hybridoma deposited as Alpha5/beta1 7H5.4.2.8 (ATCC No. PTA-7421) or the hybridoma deposited as Alpha5/beta1 7H12.5.1.4 (ATCC No. PTA-7420) in the ATCC on Mar. 7, 2006. 
     
     
         21 . (canceled) 
     
     
         22 . A composition comprising the antibody of any one of  claims 1 ,  3  or  4  and a pharmaceutically acceptable carrier. 
     
     
         23 . A method of detecting alpha5beta1 protein in sample from a patient by contacting the antibody according to any one of  claims 1 ,  3 , or  4  to the sample and detecting the anti-alpha5beta1 antibody bound to the alpha5beta1 protein. 
     
     
         24 - 26 . (canceled) 
     
     
         27 . A method for inhibiting angiogenesis and/or vascular permeability in a subject suffering from a disease having abnormal angiogenesis or vascular permeability, the method comprising administering a VEGF antagonist and an alpha5beta1 antagonist. 
     
     
         28 . The method of  claim 27 , wherein the disease is selected from the group consisting of: cancer, an ocular disease, and an autoimmune disease. 
     
     
         29 - 30 . (canceled) 
     
     
         31 . The method according to  claim 27 , wherein the subject is administered the VEGF antagonist and subsequently administered the alpha5beta1 antagonist. 
     
     
         32 . The method according to  claim 27 , wherein the subject is administered the VEGF antagonist and the alpha5beta1 antagonist concurrently. 
     
     
         33 - 34 . (canceled) 
     
     
         35 . The method according to  claim 27 , where the subject is treated with the VEGF antagonist until the subject is unresponsive to VEGF antagonist treatment and then the subject is treated with an alpha5beta1 antagonist. 
     
     
         36 . The method according to  claim 27 , wherein the disease is cancer and the subject is treated with the VEGF antagonist when the cancer is non-invasive and treated with the alpha5beta1 antagonist when the cancer is invasive. 
     
     
         37 . The method according to  claim 27 , wherein the subject has elevated alpha5beta1 levels in a diseased tissue compared to tissue from a subject not suffering from the disease. 
     
     
         38 . The method of  claim 27 , wherein the subject is further administered a therapeutic agent selected from the group consisting of an anti-neoplastic agent, a chemotherapeutic agent, a growth inhibitory agent and a cytotoxic agent. 
     
     
         39 . The method according to  claim 27 , wherein the anti-VEGF antibody can be competitively inhibited from binding to human VEGF by the Avastin® antibody. 
     
     
         40 . The method according to  claim 27 , wherein the VEGF antagonist is an anti-VEGF antibody. 
     
     
         41 . The method according to  claim 27 , wherein the alpha5beta1 antagonist is conjugated to a cytotoxic agent. 
     
     
         42 . The method according to  claim 41 , wherein the cytotoxic agent is a radioactive isotope, chemotherapeutic agent or a toxin. 
     
     
         43 . The method according to  claim 40 , wherein the anti-VEGF antibody is the Avastin® antibody. 
     
     
         44 . The method according to  claim 27 , wherein the alpha5beta1 antagonist is an antibody. 
     
     
         45 . The method according to  claim 40 , wherein the anti-VEGF antibody is a humanized or a human antibody. 
     
     
         46 . The method according to  claim 44 , wherein the anti-alpha5beta1 antibody is a humanized or a human antibody. 
     
     
         47 . A composition comprising a VEGF antagonist, an alpha5beta1 antagonist and a pharmaceutically acceptable inhibitor. 
     
     
         48 . A kit for detecting alpha5beta1 in a subject who has been treated with a VEGF antagonist, the kit comprising
 an anti-alpha5beta1 antibody produced by a hybridoma selected from the group consisting of the hybridoma deposited as Alpha5/beta1 7H5.4.2.8 (ATCC No. PTA-7421) and the hybridoma deposited as Alpha5/beta1 7H12.5.1.4 (ATCC No. PTA-7420) in the ATCC on Mar. 7, 2006, and   instructions for detecting alpha5beta1 in a subject who has been treated with a VEGF antagonist.   
     
     
         49 - 60 . (canceled)