Biosynthetic heparin

Abstract

The present disclosure relates to synthesis of heparin, which may be bioequivalent to porcine USP Heparin Sodium. The synthesis may involve three intermediates starting from heparosan.

What is claimed is: 
     
         1 . A glycosaminoglycan comprising 31-73% of NS2S6S disaccharide group; 6-40% of NS6S disaccharide group; 0-27% of NS2S group and 1-22% of NS group. 
     
     
         2 . The glycosaminoglycan of  claim 1 , comprising 36-70% of NS2S6S disaccharide group and 6-32% of NS6S disaccharide group. 
     
     
         3 . The glycosaminoglycan of  claim 1 , comprising 40-67% of NS2S6S disaccharide group and 6-26% of NS6S disaccharide group. 
     
     
         4 . The glycosaminoglycan of  claim 1 , comprising 43-64% of NS2S6S disaccharide group and 6-22% of NS6S disaccharide group. 
     
     
         5 . The glycosaminoglycan of  claim 1  comprising 12-27% of NS2S group and 1-17% of NS group. 
     
     
         6 . The glycosaminoglycan of  claim 1 , having a weight average molecular weight properties appropriate to form a final heparin product of 15,000-19,000 Da, the percentage of heparin chains with a molecular weight of greater than 24,000 Da being not more than 20% of the total, and the ratio of chains between molecular weights of 8,000 to16,000 Da to the percentage of chains with molecular weights of 16,000 to 24,000 Da being not less than 1.0. 
     
     
         7 . A method for producing a biosynthetic heparin, comprising;
 (a) obtaining a glycosaminoglycan comprising 31-73% of NS2S6S disaccharide group, 6-40% of NS6S disaccharide group, 0-27% of NS2S group and 1-22% of NS group;   (b) treating the glycosaminoglycan with an enzyme, which is 3-O-sulfotransferase isoform 1 (3OST-1), in the presence of a sulfate donor.   to produce a biosynthetic heparin batch.   
     
     
         8 . The method of  claim 7 , wherein in step (b) sufficient glucosamine residues are converted to 3-O-sulfoglucosamine residues so that the produced batch has anti-Xa and anti-IIa activities of no less than 180 units/mg and an anti-Xa/anti-IIa ratio of 0.85-1.15. 
     
     
         9 . The method of  claim 8 , wherein the anti-Xa/anti-IIa ratio is 0.9-1.1. 
     
     
         10 . The method of  claims 7 , wherein the sulfate donor comprises PAPS. 
     
     
         11 . The method of  claim 10 , wherein the PAPS is in a solution. 
     
     
         12 . The method of  claims 7 , wherein the sulfate donor comprises PAPS and PNPS. 
     
     
         13 . The method of  claims 7 , wherein said treatment is performed in the presence of a recycling system comprising PAPS, PNPS and a catalyst. 
     
     
         14 . The method of  claim 13 , wherein the catalyst is aryl sulfotransferase IV (AST-IV). 
     
     
         15 . The method of  claims 7 , wherein the enzyme is in solution. 
     
     
         16 . The method of  claims 7 , wherein the enzyme is immobilized.