Antibody-endostatin fusion protein and its variants

Abstract

Methods of inhibiting the growth of tumors comprising administering chimeric fusion molecules comprising endostatin mutants and all or a portion of anti-Her2 or anti-EGFR antibodies.

What is claimed is: 
     
         1 . A method of inhibiting the formation or growth of blood vessels in a tumor comprising the steps of administering to an individual who has been diagnosed with a tumor, a therapeutically effective amount of a pharmaceutical composition comprising an isolated chimeric fusion molecule, wherein the chimeric fusion molecule comprises an anti-tumor antigen binding domain from an anti-HER2 or anti-EGFR antibody and at least one human endostatin protein or fragment thereof, wherein the at least one human endostatin protein or fragment thereof comprises a proline to alanine amino acid substitution at position 125 of human endostatin, wherein administration of the pharmaceutical composition results in reduction in the blood vessels in the tumor. 
     
     
         2 . The method of  claim 1 , wherein the antigen binding domain specifically binds one or more tumor antigens. 
     
     
         3 . The method of  claim 1 , wherein the human endostatin protein or fragment thereof comprise one or more NGR motifs (Asn-Gly-Arg) and/or RGD (Arg-Gly-Asp) motifs. 
     
     
         4 . The method of  claim 3 , wherein the one or more NGR motifs (Asn-Gly-Arg) and RGD (Arg-Gly-Asp) motifs are located at the amino (NH 2 —) terminal, and/or carboxy terminal (COOH—) and/or amino acid positions 93-133 of human endostatin protein or fragment thereof. 
     
     
         5 . The method of  claim 3 , wherein the one or more NGR motifs (Asn-Gly-Arg) and RGD (Arg-Gly-Asp) motifs are located at amino acid positions 126-128 following the proline or alanine at position 125 of the human endostatin protein or fragment thereof. 
     
     
         6 . The method of  claim 1 , wherein the antibody or fragment thereof, is IgA, IgM, IgG, IgE, or IgD. 
     
     
         7 . The method of  claim 1 , wherein the chimeric fusion protein is administered to a patient, simultaneously and/or in separate treatments with one or more of: cytoximab, sunitinib, sorafenib, celebrex, MTOR inhibitors, AKT inhibitors, P13K inhibitors, bevacizumab (Avastin), signal transduction inhibitors, tamoxifen, toremifen, raloxifene, droloxifene, iodoxyfene, megestrol acetate, anastrozole, letrazole, borazole, exemestane, flutamide, nilutamide, bicalutamide, cyproterone acetate, goserelin acetate, luprolide, finasteride, herceptin, methotrexate, 5-fluorouracil, cytosine arabinoside, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin, mithramycin, cisplatin, carboplatin, melphalan, chlorambucil, busulphan, cyclophosphamide, ifosfamide, nitrosoureas, thiotephan, vincristine, taxol, taxotere, etoposide, teniposide, amsacrine, irinotecan, topotecan, an epothilone; a tyrosine kinase inhibitor, Iressa or OSI-774; an angiogenesis inhibitor; an EGF inhibitor; a VEGF inhibitor; a CDK inhibitor; a Her1/2 inhibitor and monoclonal antibodies directed against growth factor receptors. 
     
     
         8 . The method of  claim 1 , wherein the chimeric fusion protein is administered in combination with and/or in separate treatments, one or more antibodies comprising cytoximab, sunitinib, sorafenib, celebrex, MTOR inhibitors, AKT inhibitors, P13K inhibitors, bevacizumab (Avastin), signal transduction inhibitors, anti PDL1, anti CTLA4, and anti her2 antibodies. 
     
     
         9 . The method of  claim 1 , wherein the chimeric fusion protein is administered in combination with and/or in separate treatments, one or more anti-angiogenic factors comprising sunitinib, sorafenib, and angiostatin. 
     
     
         10 . A method of identifying treatment option for an individual diagnosed with a tumor comprising the steps of:
 a) obtaining a biopsy sample of the tumor;   b) determining if a chimeric fusion molecule comprising an anti-tumor antigen binding domain from an anti-HER2 antibody or anti-EGFR antibody and at least one human endostatin protein or fragment thereof, wherein the at least one human endostatin protein or fragment thereof comprises a proline to alanine amino acid substitution at position 125 of human endostatin, inhibits vasculogenic mimicry in the tumor cells; and   c) if inhibition of vasculogenic mimicry is observed, then identifying the chimeric fusion molecule as a useful treatment option for the individual.   
     
     
         11 . The method of  claim 10 , comprising a further step of determining if the tumor cells exhibit vasculogenic mimicry between steps a) and b). 
     
     
         12 . A method of treatment of an individual diagnosed with a tumor comprising the steps of combining:
 a) removing the tumor using surgical or radiation treatment; and   b) administration of a chimeric fusion molecule comprising an anti-tumor antigen binding domain from an anti-HER2 antibody or anti-EGFR antibody and at least one human endostatin protein or fragment thereof, wherein the at least one human endostatin protein or fragment thereof comprises a proline to alanine amino acid substitution at position 125 of human endostatin,
 wherein the removing the tumor is done before, during or after the administration of the chimeric fusion molecule.