Method of producing biologically active human acidic fibroblast growth factor and its use in promoting angiogenesis

Abstract

The present invention relates to the treatment of coronary heart disease by revascularization therapy, and more particularly to the intramyocardial injection of a pharmaceutical composition comprising a recombinant fibroblast growth factor-1 protein or a fragment of a recombinant fibroblast growth factor-1 protein, optionally, with a physiologic glue for inducing local neoangiogenesis in ischemic myocardium. Methods of producing the recombinant fibroblast growth factor 1 protein and fragments are also disclosed.

1 . A method for revascularizing an ischemic region, comprising the steps of:
 (a) preparing a pharmaceutical composition comprising a recombinant fibroblast growth factor-1 (FGF-1); and   (b) injecting an amount of said pharmaceutical composition into the ischemic region, said amount being sufficient to induce local neoangiogenesis.   
     
     
         2 . The method of  claim 1 , wherein the expressible gene has a sequence which is contained within the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID NO: 6. 
     
     
         3 - 11 . (canceled) 
     
     
         12 . The method of  claim 1 , wherein the human acidic fibroblast growth factor protein has the sequence as set forth in SEQ ID NO: 7. 
     
     
         13 - 15 . (canceled) 
     
     
         16 . The method of  claim 1 , wherein the biologically active human acidic fibroblast growth factor protein contains amino acids 9-155 as shown in SEQ ID NO: 2. 
     
     
         17 . The method of  claim 1 , wherein the biologically active human acidic fibroblast growth factor protein contains amino acids 2-141 as shown in SEQ ID NO: 7. 
     
     
         18 . The method of  claim 1 , wherein the biologically active human acidic fibroblast growth factor protein contains amino acids 2-135 as shown in SEQ ID NO: 5. 
     
     
         19 . The method of  claim 1 , wherein the biologically active human acidic fibroblast growth factor protein comprises a sequence shown in SEQ ID NO: 8. 
     
     
         20 . The method of  claim 1 , wherein said FGF-1 is injected at a final concentration in a range of about 0.1 μg/kg body weight per site to about 10 mg/kg body weight per site. 
     
     
         21 . The method of  claim 1  wherein said FGF-1 is injected at a final concentration in a range of about 10 to 100 μg/kg body weight per site. 
     
     
         22 . The method of  claim 1 , wherein the pharmaceutical composition further comprises a physiologic glue. 
     
     
         23 . The method of  claim 22 , wherein said physiologic glue is fibrin glue. 
     
     
         24 . The method of  claim 1 , wherein said FGF-1 and said physiologic glue are mixed immediately prior to application. 
     
     
         25 . The method of  claim 1 , wherein said pharmaceutical composition further comprises an anticoagulant. 
     
     
         26 . The method  claim 25 , wherein said anticoagulant is heparin. 
     
     
         27 . The method of  claim 26 , wherein the heparin is applied at a final concentration in a range of about 1 Upper ml to about 1000 Upper ml. 
     
     
         28 . The method of  claim 1 , wherein said injecting step further comprises:
 making a thoracotomy incision;   identifying the at least one site of coronary artery stenosis;   administering a 0-blocker to reduce the heart rate to a range of about 20-60 beats per minute; and   injecting the pharmaceutical composition intramyocardially at or near the at least one site of coronary artery stenosis.   
     
     
         29 . The method of  claim 28 , wherein said thoracotomy incision further comprises an anterior left-sided incision; dissecting a region of costal cartilage over a 5 th  rib; and
 opening a left pleural space and a pericardium.   
     
     
         30 . The method of  claim 28 , wherein the step of identifying the at least one site of coronary artery stenosis further comprises retracting the heart forward using traction sutures. 
     
     
         31 . The method of  claim 1 , wherein the neoangiogenesis is long term and occurs in the ischemic region at 6 weeks after the injection. 
     
     
         32 . The method of  claim 1 , wherein the neoangiogenesis is long term and occurs in the ischemic region at 3 months after the injection. 
     
     
         33 . The method of  claim 1 , wherein the method further comprises performing a coronary artery bypass graft. 
     
     
         34 . The method of  claim 1 , further comprising the step of injecting a composition comprising a physiologic glue subsequent to injection with the pharmaceutical composition. 
     
     
         35 . A method for treating coronary artery disease in a patient, comprising the steps of:
 (a) preparing a pharmaceutical composition comprising a recombinant fibroblast growth factor-1 (FGF-1);   (b) injecting an amount of said pharmaceutical composition into at least one site in a heart wall, said amount being sufficient to improve myocardial perfusion; and   (c) injecting a composition comprising a physiological glue to a surface of the heart at the site(s) where the pharmaceutical composition was injected.