Pharmaceutical compositions including variants of vascular endothelial cell growth factor having altererd pharmacological properties

Abstract

Described herein are pharmaceutical compositions including vascular endothelial cell growth factor (VEGF) variants having modifications in the C-terminal heparin binding domain. The variants exhibit reduced clearance rates for systemic administration generally at lower doses compared with native VEGF thus providing variants having longer availability for therapeutic effect.

1 . An isolated DNA sequence comprising a sequence that encodes a vascular endothelial cell growth factor (VEGF) variant wherein said variant comprises C-terminus heparin binding domain modifications.  
     
     
         2 . The DNA sequence according to  claim 1  wherein said DNA sequence encodes a variant containing a C-terminus modification beyond about amino acid 120.  
     
     
         3 . A polypeptide which comprises a vascular endothelial cell growth factor variant comprising a C-terminus heparin binding domain structural alteration resulting in modified pharmacokinetic properties.  
     
     
         4 . A polypeptide according to  claim 3  wherein said C-terminus heparin binding domain contains cleaved sites therein.  
     
     
         5 . A polypeptide according to  claim 3  wherein said C-terminus heparin binding domain contains amino acid variations compared with native vascular endothelial cell growth factor.  
     
     
         6 . A polypeptide according to  claim 5  wherein said variations comprise a reduction in the effect of positively charged amino acids.  
     
     
         7 . A polypeptide according to  claim 6  wherein said variant has altered C-terminus heparin binding domain conformational structure.  
     
     
         8 . A polypeptide which comprises a vascular endothelial cell growth factor variant containing a C-terminus heparin binding domain modification such that the binding characteristic of said domain is altered resulting in said polypeptide having a reduced clearance rate compared with native VEGF.  
     
     
         9 . A replicable expression vector capable in a transformant host cell of expressing the DNA sequence of  claim 1 .  
     
     
         10 . Host cells transformed with a vector according to  claim 9 .  
     
     
         11 . Host cells according to  claim 10  which are Chinese hamster ovary cells.  
     
     
         12 . A composition of matter comprising the VEGF variant according to  claim 3  or  8  compounded with a pharmaceutically acceptable carrier.  
     
     
         13 . A method of treatment which comprises administering a composition according to  claim 12 .  
     
     
         14 . A method of preparing a VEGF variant according to  claim 4  which comprises expressing transforming DNA encoding said variant in a recombinant host cell.  
     
     
         15 . A method for preparing a polypeptide according to  claim 3  which comprises introducing cleavage sites within the C-terminus heparin binding domain.  
     
     
         16 . A method according to  claim 14  wherein said variant contains fewer C-terminus heparin binding domain amino acids exhibiting a positive charge.  
     
     
         17 . A method according to  claim 14  wherein said variant contains deletions in the C-terminus heparin binding domain.  
     
     
         18 . A method according to  claim 14  wherein said variant contains amino acid substitutions in the C-terminus heparin binding domain.  
     
     
         19 . A polypeptide according to  claim 3  or  8  having amino acids 1 to 120 of native vascular endothelial cell growth factor.  
     
     
         20 . A vascular endothelial cell growth factor having a reduced in vivo clearance rate compared with native vascular endothelial cell growth factor.