Fucosylated chondroitin sulfate as a new oral antithrombotic agent

Summary

Fucosylated chondroitin sulfate is a potent anticoagulant polysaccharide extracted from sea cucumber. Its anticoagulant activity is attributed to the presence of sulfated fucose branches. We have shown that intravascular injection of fucosylated chondroitin sulfate inhibits thrombus formation in a venous and an arterial shunt model in rats. Since this compound resists digestion by enzymes that cleave mammalian glycosaminoglycans, we investigated the possibility that fucosylated chondroitin sulfate might be absorbed after oral administration. In fact, after oral administration of fucosylated chondroitin sulfate to rats, we observed a dose-dependent increase in the plasma anticoagulant activity, as assessed by assays for activated partial thromboplastin time (aPTT) and thrombin time (TT) (about 3-and 5-fold, respectively) and by anti-IIa activity. Furthermore, animals receiving daily oral doses of this glycosaminoglycan showeda decrease in thrombus weight on experimental models of venous and arterial shunt thrombosis. This antithrombotic action clearly has a strong relationship with anticoagulant activity. Similar doses of heparin administered orally had no effect on the plasma anticoagulant activity or on the thrombus weight. Finally, we observed that fucosylated chondroitin sulfate given orally to rats did not modify the bleeding time. Overall, our results indicate that fucosylated chondroitin sulfate is absorbed after oral administration and could become a promising oral anticoagulant.

Keywords

Glycosaminoglycans - fucosylated chondroitin sulfate - oral anticoagulants - heparin

• References

• 1 Biermann CH, Marks JA, Vilela-Silva AC. et al. Carbohydrate-based species recognition in sea urchin fertilization: another avenue for speciation?. Evol Dev 2004; 06: 353-61.
• 2 Caterson B, Mahmoodian F, Sorrel JM. et al. Modulation of native chondroitin sulfate structures in tissue development and disease. J Cell Sci 1990; 97: 411-7.
• 3 Mourão PAS, Pereira MS. Searching for alternatives to heparin. Trends Cardiovasc Med 1999; 09: 225-32.
• 4 Mulloy B, Mourão PAS, Gray E. Structure/function studies of anticoagulant sulfated polysaccharides using NMR. J Biotech 2000; 77: 123-35.
• 5 Mourão PAS, Guimarães B, Mulloy B. et al. Antithrombotic activity of a fucosylated chondroitin sulfate from echinoderm: Sulfated fucose branches on the polysaccharide account for its antithrombotic action. Br J Haematol 1998; 101: 647-52.
• 6 Pacheco RG, Vicente CP, Zancan P. et al. Different antithrombotic mechanisms among glycosaminoglycans revealed witha new fucosylated chondroitin sulfate from an echinoderm. Blood Coagul Fibrinolysis 2000; 11: 563-73.
• 7 Zancan P, Mourão PAS. Venous and arterial thrombosis in rat models: dissociation of the antithrombotic effects of glycosaminoglycans. Blood Coagul Fibrinolysis 2004; 15: 45-54.
• 8 Vieira RP, Mourão PAS. Occurrence of a unique fucose-branched chondroitin sulfate in the body wall of a sea cuccumber. J Biol Chem 1988; 263: 18176-83.
• 9 Vieira RP, Mulloy B, Mourão PAS. Structure of a fucose-branched chondroitin sulfate from sea cuccumber: evidence for the presence of 3-O-sulfo-β-Dglucuronosyl residues. J Biol Chem 1991; 266: 13530-6.
• 10 Vinazzer H. Assay of total factor XII and of activated factor XII in plasma with a chromogenic substrate. Thromb Res 1979; 14: 155-166.
• 11 Wessler S, Reimer SM, Sheps MC. Biological assay of a thrombosis inducing activity in human serum. J Appl Physiol 1959; 14: 943-6.
• 12 Umetsu T, Sanai K. Effect of 1-methyl-2-mercapto-5-(3-pyridyl)-imidazole (KC-6141), an anti-aggregating compound, on experimental thrombosis in rats. Thromb Haemost 1978; 39: 74-83.
• 13 Herbert JM, Bernat A, Maffrand JP. Importance of platelets in experimental venous thrombosis in rats. Blood 1992; 80: 2281-6.
• 14 Mourão PAS, Boisson-Vidal C, Tapon-Bretaudiere J. et al. Inactivation of thrombin bya fucosylated chondroitin sulfate from echinoderm. Thromb Res 2001; 102: 167-76.
• 15 Zhiguang LI, Hongli W, Jiazeng LI. et al. Basic and clinical study on the antithrombotic mechanism of glycosaminoglycan extracted from sea cucumber. Chin Med J 2000; 113: 706-11.
• 16 Yoon SJ, Pereira MS, Pavão MSG. et al. The medicinal plant Porana volubilis contains polysaccharides with anticoagulant activity mediated by heparin cofactor II. Thromb Res 2002; 106: 51-8.
• 17 Rocha HAO, Moraes FA, Trindade ES. et al. Structural and hemostatic activities of a sulfated galactofucan from the brown alga Spatoglossum schroederi: an ideal antithrombotic agent?. J Biol Chem 2006; 280: 41278-88.
• 18 Mourão PAS, Pereira MS, Pavão MS. et al. Structure and anticoagulant activity of a fucosylated chondroitin sulfate from echinoderm. Sulfated fucose branches on the polysaccharide account for its high anticoagulant action. J Biol Chem 1996; 271: 23973-84.
• 19 Leadley Jr RJ, Chi L, Rebello SS. et al. Contribution of in vivo models of thrombosis to the discovery and development of novel antithrombotic agents. J Pharmacol Toxicol Methods 2000; 43: 101-16.
• 20 Hu M, Hsi-Hsien L, Weng-Ching K. Hypolipidemic Effect of glycosaminoglycans from the sea cucumber Metriatyla scabra in rats fed a cholesterolsupplemented diet. J Agric Food Chem 2002; 50: 3602-6.
• 21 Imanari T, Washio T, Huang H. et al. Oral absorption and clearance of partially depolymerized fucosyl chondroitin sulfate from sea cucumber. Thromb Res 1999; 93: 259-74.
• 22 Hiebert LM, Pinel C, Wice SM. Orally administered heparins prevent arterial thrombosis in a rat model. Thromb Haemost 2004; 91: 919-26.
• 23 Hiebert LM, Wice SM, Ping T. et al. Antithrombotic efficacy ina rat model of the low molecular weight heparin, reviparin sodium, administered by the oral route. Thomb Haemost 2001; 85: 114-8.
• 24 Ronca G, Ronca F, Palmieri L. Anti-inflammatory activity of chondroitin sulfate. Osteoarthrits Cart 1998; 06 Suppl A 14-21.
• 25 Volpi N. Oral bioavailability of chondroitin sulfate (Condrosulf ®) and its constituents in healthy male volunteers. Osteoarthrits Cart 2002; 10: 768-77.
• 26 Wuisman P. Glucosamine and chondroitin sulfate supplementation to treat symptomatic disc degeneration: Biochemical rationale and case report. BMC Compl Altern Med 2003; 03: 1-8.
• 27 Houin G, Barthe L, Woodley J. et al. In vitro intestinal degradation and absorption of chondroitin sulfate, a glycosaminoglycan drug. Arzneimittelforschung 2004; 54: 286-92.
• 28 Gomez-Orellana I, Goldberg M. Challenges for the oral delivery of macromolecules. Nat Rev 2003; 02: 289-95.
• 29 Hirsh J, Levine LM. Low molecular weight heparin. Blood 1992; 79: 1-18.
• 30 Perzbone E, Strassburger J, Wilmer A. et al. In vitro and in vivo studies of the novel antithrombotic agent BAY 59-7939: an oral, direct Factor Xa inhibitor. Thromb Haemost 2005; 03: 514-21.
• 31 Gustafsson D, Bylund R, Antosson T. et al. A new oral anticoagulant: the 50-year challenge. Nat Rev 2004; 03: 649-59.