Regioselectivity in the sulfation of dermatan sulfate and methyl 4,6-O-benzylidene-α-d-idopyranoside

Abstract The sulfation of dermatan sulfate by SO 3 -trimethylamine in N,N -dimethylformamide led to substitution initially at HO-6 of residues of 2-acetamido-2-deoxy-β- d -galactopyranosyl 4-sulfate ( 1 ), to produce the 4,6-disulfate ( 6 ). When this step reached a level of > 50%, sulfation occurred with equal facility at HO-2 and HO-3 of residues of α- l -idopyranosyluronic acid ( 2 ), giving rise to a mixture of 2-,3-, and 2,3-disulfates. An analogous substitution pattern was observed for HO-2 and -3 of a simpler idopyranose unit, in the sulfation of methyl 4,6- O -benzylidene-α- d -idopyranoside ( 12 ). This lack of regioselectivity in the reaction of 2 (and 12 ) contrasts markedly with the high affinity of the reagent for HO-3 of residues of α- l -idopyranosyluronic acid present in a modified form of heparin. It is attributed to a difference between the two polymers in the relative orientation of their neighboring amino sugar residues, whereby there is an unobstructed access of the reagent in one instance, and hindrance of HO-2 selectively in the other. Enzymolysis by chondroitinase ABC was found to yield unsaturated disaccharide containing residues of 4,6-disulfate, as well as larger fragments containing unsaturated glycosyl groups derived from l -idopyranosyluronic acid 2-sulfate, evidence of a relatively broad enzyme specificity. The presence of extra sulfate groups in dermatan sulfate did not enhance its weak antithrombotic activity, as measured by anti Xa assay, in disagreement with earlier reports.