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

Thromb Res. 2010 Sep;126(3):e180-7. doi: 10.1016/j.thromres.2010.04.008. Epub 2010 May 31.

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, and 13C NMR spectroscopy. Its anticoagulant activity was determined by assays of aPTT, TT, alpha-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 beta-D-Manp units and OH-2 and OH-4 of the (1-->6)-linked alpha-D-Galp 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 alpha-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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anticoagulants / isolation & purification
  • Anticoagulants / pharmacology*
  • Anticoagulants / toxicity
  • Antithrombin Proteins / metabolism*
  • Bleeding Time
  • Blood Coagulation / drug effects*
  • Chromatography, Gel
  • Disease Models, Animal
  • Dose-Response Relationship, Drug
  • Factor Xa / metabolism
  • Factor Xa Inhibitors
  • Female
  • Fibrinolytic Agents / isolation & purification
  • Fibrinolytic Agents / pharmacology*
  • Fibrinolytic Agents / toxicity
  • Galactans / isolation & purification
  • Galactans / pharmacology*
  • Galactans / toxicity
  • Hemorrhage / chemically induced
  • Heparin Cofactor II / metabolism*
  • Humans
  • Magnetic Resonance Spectroscopy
  • Male
  • Partial Thromboplastin Time
  • Platelet Aggregation / drug effects
  • Pleurotus / chemistry
  • Prothrombin Time
  • Rats
  • Rats, Wistar
  • Scattering, Radiation
  • Structure-Activity Relationship
  • Sulfates / isolation & purification
  • Sulfates / pharmacology*
  • Sulfates / toxicity
  • Thrombin / antagonists & inhibitors*
  • Thrombin / metabolism
  • Venous Thrombosis / blood
  • Venous Thrombosis / prevention & control

Substances

  • Anticoagulants
  • Antithrombin Proteins
  • Factor Xa Inhibitors
  • Fibrinolytic Agents
  • Galactans
  • Sulfates
  • Heparin Cofactor II
  • Thrombin
  • Factor Xa