Conjugates of carboxy polysaccharides with fibroblast growth factors and variants thereof

Abstract

Conjugates of hydrophilic carboxy polysaccharides with protein members of the fibroblast growth factor (FGF) family and variants thereof. The conjugates provide a modified bioactivity and stability of FGFs for various therapeutic applications.

1 .- 14 . (canceled) 
     
     
         15 . A conjugate comprising a fibroblast growth factor (FGF) or a variant thereof covalently coupled to a hyaluronic acid, wherein said FGF is selected from FGF-2 and FGF-18. 
     
     
         16 . The conjugate of  claim 15 , wherein the molecular weight of said hyaluronic acid is 17 kDa to 234 kDa. 
     
     
         17 . The conjugate of  claim 15 , wherein the molar ratio of FGF to said hyaluronic acid is within the range of 1:1 to 50:1. 
     
     
         18 . The conjugate according to  claim 15 , wherein the FGF variant is an N-terminal truncated variant. 
     
     
         19 . The conjugate according to  claim 15 , formed by coupling the amino functional group of FGF or variant thereof with an active ester carboxy moiety of the hyaluronic acid. 
     
     
         20 . The conjugate of  claim 15 , wherein the hyaluronic acid and the FGF or the variant thereof are directly coupled via an amide bond between a carboxylic functional group of hyaluronic acid and an amino functional group of said FGF or a variant thereof. 
     
     
         21 . The conjugate of  claim 15 , wherein the conjugate exerts activity of the FGF or variant thereof which is at least 80% of the activity of a non-conjugated FGF or variant thereof. 
     
     
         22 . The conjugate of  claim 15 , wherein the conjugate exerts activity of the FGF or variant thereof which is longer by at least 1 day than the activity of the non-conjugated FGF or variant thereof. 
     
     
         23 . The conjugate of  claim 15 , wherein the conjugate exerts activity of the FGF or variant thereof for at least 7 days. 
     
     
         24 . The conjugate of  claim 15 , wherein said conjugated FGF has a prolonged activity in vivo by at least two fold, when compared to the non-conjugated FGF or variant thereof. 
     
     
         25 . The conjugate of  claim 15 , wherein the conjugate is active in the conjugate form. 
     
     
         26 . The conjugate of  claim 15 , wherein the FGF or the variant thereof and the hyaluronic acid are covalently coupled via a linker comprising a hydrazido moiety coupled to a maleimido moiety or a vinyl sulfone moiety. 
     
     
         27 . The conjugate of  claim 26 , wherein the conjugate is inactive in the conjugate form. 
     
     
         28 . The of  claim 15 , further comprising fibrin(ogen), wherein the fibrin(ogen) and FGF or variant thereof are each coupled directly with the hyaluronic acid via an amide bond between a carboxylic functional group of the hyaluronic acid and an amino functional group of said fibrin(ogen) and FGF or variant thereof, thereby forming a triple conjugate. 
     
     
         29 . The conjugate of  claim 28 , wherein the fibrinogen is cleaved by a fibrinogen cleaving agent, thereby producing a water insoluble fibrin clot. 
     
     
         30 . The conjugate of  claim 29 , wherein the water insoluble clot is freeze-dried, thereby producing a porous fibrin matrix or scaffold. 
     
     
         31 . A pharmaceutical composition comprising a conjugate according to  claim 15  and a pharmaceutically acceptable carrier or excipient.