Antithrombin and heparin cofactor II-mediated inactivation of α-thrombin by a synthetic, sulfated mannogalactan

Abstract Introduction A mannogalactan from Pleurotus ostreatoroseus (MgPr) was chemically sulfated to give MgPr-S1, which was evaluated for its anticoagulant and antithrombotic activities, bleeding tendency, and platelet aggregation. Materials and methods MgPr-S1 was partially characterized by HPSEC-MALLS, methylation analysis, and13 C NMR spectroscopy. Its anticoagulant activity was determined by assays of aPTT, TT, α-thrombin and factor Xa residual activity, heparin cofactor II (HCII)-, or antithrombin (AT)-mediated inhibition. The antithrombotic effect was evaluated in rats using a venous thrombosis model and the bleeding tendency was also tested in vivo. Platelet aggregation was investigated by an adaptation of the method of Born [1]. Results The hydroxyl groups of β-D-Man p units and OH-2 and OH-4 of the (1→6)-linked α-D-Gal p units were preferentially substituted. The anticoagulant activity of MgPr-S1 was mainly by thrombin inhibition with antithrombin and HCII, and had an effect on platelet aggregation induced by ADP and α-thrombin. It almost completely inhibited thrombus formation in vivo at a dose of 6 mg/kg and heparin inhibited thrombus formation at a dose of 0.200 mg/kg. Conclusions These results suggested that the chemically sulfated mannogalactan could act as an alternative to heparin as anticoagulant.