Method of Promoting Natural Bypass

Abstract

An angiogenic factor comprising a mixture of proteins derived from bone. The angiogenic protein mixture is produced by a series of steps that allow the proteins to be kept in solution. The angiogenic mixture of bone proteins is produced by a multi-step process that includes at least one ultrafiltration step, an anion exchange chromatography step, a cation exchange chromatography step and a high performance liquid chromatography (HPLC) purification step.

1 - 37 . (canceled)  
   
   
       38 . A method of improving muscle blood flow in a mammal, comprising administering to the mammal a mixture of proteins derived from ground bone.  
   
   
       39 . The method according to  claim 38 , wherein the mixture of proteins derived from ground bone comprises at least two growth factors selected from the group consisting of bone morphogenic protein-2 (BMP-2), bone morphogenic protein-3 (BMP-3), bone morphogenic protein-4 (BMP-4), bone morphogenic protein-5 (BMP-5), bone morphogenic protein-6 (BMP-6), bone morphogenic protein-7 (BMP-7), transforming growth factor β1 (TGF-β1), transforming growth factor β2 (TGF-β2), transforming growth factor β3 (TGF-β3) and fibroblast growth factor 1 (FGF-1).  
   
   
       40 . The method of  claim 38 , wherein the mammal is a human.  
   
   
       41 . The method of  claim 38 , wherein the mixture is administered subcutaneously, intramuscularly, or intravenously.  
   
   
       42 . The method of  claim 38 , wherein the mixture is administered discretely or continuously.  
   
   
       43 . The method of  claim 38 , wherein said mixture further comprises a growth factor selected from the group consisting of insulin-like growth factor-1 (IGF-1), epidermal growth factor (EGF), hepatocyte growth factor (HGF), transforming growth factor α (TGF-α), and platelet-derived growth factor (PDGF).  
   
   
       44 . The method of  claim 38 , wherein the mixture further comprises a preservative or an adjuvant.  
   
   
       45 . The method of  claim 38 , wherein the bone is mammalian bone.  
   
   
       46 . The method of  claim 45 , wherein the mammalian bone is bovine bone.  
   
   
       47 . The method of  claim 38 , wherein the mixture is derived by: 
 (i) grinding mammalian bone, to produce ground bone;    (ii) cleaning the ground bone, to produce cleaned ground bone;    (iii) demineralizing the cleaned ground bone, to produce demineralized cleaned ground bone;    (iv) extracting protein from the demineralized cleaned ground bone using a protein denaturant to yield extracted protein;    (v) ultrafiltering the extracted protein to separate out high molecular weight proteins;    (vi) ultrafiltering the extracted protein to separate out low molecular weight proteins;    (vii) transferring the extracted protein to a non-ionic denaturant;    (viii) subjecting the extracted protein to an anion exchange process;    (ix) subjecting the extracted protein to a cation exchange process; and    (x) subjecting the extracted protein to a reverse phase HPLC process.    
   
   
       48 . The method of  claim 38 , wherein the muscle is myocardial muscle.  
   
   
       49 . The method of  claim 38 , wherein the muscle is ischemic.  
   
   
       50 . The method of  claim 49 , wherein the muscle is served by an at least partially occluded vessel.  
   
   
       51 . The method of  claim 38 , wherein the mixture of proteins comprises BMP-2, BMP-3, BMP-7, TGF-β, and FGF-1.  
   
   
       52 . The method of  claim 38 , wherein the mixture of proteins is mixed with a carrier selected from the group consisting of polylactic acid, polyglycolic acid, copolymers of lactic acid and glycolic acid, collagen, polyalkylene ether copolymer surfactant, and polyvinylpyrrolidone.  
   
   
       53 . The method of  claim 52 , wherein the carrier is polyvinylpyrrolidone.  
   
   
       54 . The method of  claim 53 , wherein the mixture of the proteins and the carrier contains 10 μg of the mixture of proteins per 0.1 cc of polyvinylpyrrolidone or 100 μg of the mixture of proteins per 0.1 cc of polyvinylpyrrolidone.