Methods of treating bone disease using vascular endothelial growth factor fusion constructs

Abstract

The 121-amino acid isoform of vascular endothelial growth factor (VEGF 121 ) is linked by a flexible G4S tether to a cytotoxic molecule such as toxin gelonin or granzyme B and expressed as a soluble fusion protein. The VEGF 121 fusion protein exhibits significant anti-tumor vascular-ablative effects that inhibit the growth of primary tumors and inhibit metastatic spread and vascularization of metastases. The VEGF 121 fusion protein also target osteoclast precursor cells in vivo and inhibits osteoclastogenesis.

1 - 21 . (canceled) 
     
     
         22 . A method of treating a subject with a bone disease, comprising administering to a subject with a bone disease a pharmaceutically effective amount of a composition comprising a conjugate comprising a cytotoxic polypeptide and a VEGF polypeptide that binds to both vascular endothelial growth factor (VEGF) receptor type 1 (Flt-1) and VEGF receptor type 2 (kinase domain receptor/Flk-1). 
     
     
         23 . The method of  claim 22 , wherein the subject is a human. 
     
     
         24 . The method of  claim 22 , wherein the bone disease is an osteolytic bone lesion. 
     
     
         25 . The method of  claim 22 , wherein the bone disease is bone lysis. 
     
     
         26 . The method of  claim 22 , wherein the bone disease is osteoporosis. 
     
     
         27 . The method of  claim 22 , wherein the bone disease is osteoarthritis. 
     
     
         28 . The method of  claim 22 , wherein the bone disease is a tumor. 
     
     
         29 . The method of  claim 22 , wherein tumor is skeletal metastasis. 
     
     
         30 . The method of  claim 22 , wherein the conjugate is a fusion protein of a VEGF polypeptide and a cytotoxic polypeptide. 
     
     
         31 . The method of  claim 22 , wherein said VEGF polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:28-34. 
     
     
         32 . The method of  claim 22 , wherein the cytotoxic polypeptide is a toxin or a signal transduction protein capable of generating apoptotic signals. 
     
     
         33 . The method of  claim 22 , wherein the toxin is gelonin. 
     
     
         34 . The method of  claim 32 , wherein the signal transduction protein capable of generating apoptotic signals is selected from the group consisting of granzyme B, Bax, TNF-a, TNF-b, TNF-like molecule, TGF-b, IL-12, IL-3, IL-24, IL-18, TRAIL, IFN-a, IFN-b, IFN-g, Bcl-2, Fas ligand and caspases. 
     
     
         35 . The method of  claim 34 , wherein the signal transduction protein capable of generating apoptotic signals is granzyme B. 
     
     
         36 . The method of  claim 22 , wherein the conjugate is a fusion protein comprising the 121-amino acid isoform of vascular endothelial growth factor (VEGF 121 ) and gelonin. 
     
     
         37 . The method of  claim 22 , wherein the conjugate is a fusion protein comprising the 121-amino acid isoform of vascular endothelial growth factor (VEGF 121 ) and granzyme B.