An algal sulfated galactan has an unusual dual effect on venous thrombosis due to activation of factor XII and inhibition of the coagulation proteases

Summary

Sulfated galactan from the red alga Botryocladia occidentalis has a potent anticoagulant activity,due to its ability to enhance thrombin and factor Xa inhibition by antithrombin and/or heparin cofactor II. It is less active than unfractionated heparin in arterial thrombosis,but in a venous thrombosis presents a dual effect,inhibiting thrombosis in low but not in high doses.This dual effect on venous thrombosis is a consequence of two actions, one that inhibits thrombin and factor Xa and one that induces factor XII activation. In order to dissociate these effects, we prepared derivatives of the sulfated galactan with low molecular weights. Two fractions that were similar in size to unfractionated heparin and low-molecular-weight heparin were obtained. As the molec- ular weight decreased, the ability to activate factor XII and to promote inhibition of coagulation proteases in the presence of antithrombin and heparin cofactor II diminished.At ∼5 kDa, the sulfated galactan fragment had no effect on factor XII activation, and showed the same effect as unfractionated heparin in a venous thrombosis model.The ∼5-kDa fragment is an antithrombotic with several advantages: i) It is as active as unfractionated heparin in venous thrombosis, but it has little activity in arterial thrombosis; ii) It inhibits venous thrombosis with very little anticoagulant effect; iii) It does not cause bleeding; and iv) It is not obtained from mammals.

Keywords

Sulfated polysaccharide - anticoagulant polysaccharide - antithrombotic polysaccharide - heparin - Hageman factor

• References

• 1 Fareed J, Hoppensteadt DA. Heparins in the new millennium: Will unfractionated heparin survive?. Semin Thromb Haemost 2000; 26: 87-88.
• 2 Beguin S, Lindhout T, Hemker HC. The effect of trace amounts of tissue factor on thrombin generation in platelet rich plasma, its inhibition by heparin. Thromb Haemost 1989; 61: 25-29.
• 3 Warkentin TE. Heparin-induced thrombocytopenia: A clinicopathologic syndrome. Thromb Haemost 1999; 82: 439-447.
• 4 Visentin GP. Heparin-induced thrombocytopenia: Molecular pathogenesis. Thromb Haemost 1999; 82: 448-456.
• 5 Kelton JG, Hirsh J. Bleeding associated with antithrombotic therapy. Semin Hematol 1980; 17: 259-291
• 6 Kakkar VV, Howes J, Sharma V. et al. A comparative, double-blind, randomised trial of a new second generation LMWH (Bemiparin) and UFH in the prevention of post-operative venous thromboembolism. Thromb Haemost 2000; 83: 523-529.
• 7 Liaw PCY, Becker DL, Stafford AR. et al. Molecular basis for the susceptibility of fibrin-bound thrombin to inactivation by heparin cofactor II in the presence of dermatan sulfate but not heparin. J Biol Chem 2001; 276: 20959-20965.
• 8 Farias WRL, Valente AP, Pereira MS. et al. Structure and anticoagulant activity of sulfated galactans – Isolation of a unique sulfated galactan from the red algae Botryocladia occidentalis and comparison of its anticoagulant action with that of sulfated galactans from invertebrates. J Biol Chem 2000; 275: 29299-29307.
• 9 Pereira MS, Melo FR, Mourão PAS. Is there a correlation between structure and anticoagulant action of sulfated galactans and sulfated fucans?. Glycobiology 2002; 12: 573-580.
• 10 Pereira MS, Mulloy B, Mourão PAS. Structure and anticoagulant activity of sulfated fucans – Comparison between the regular, repetitive, and linear fucans from echinoderms with the more heterogeneous and branched polymers from brown algae. J Biol Chem 1999; 274: 7656-7667.
• 11 Melo FR, Pereira MS, Foguel D. et al. Antithrombin- mediated anticoagulant activity of sulfated polysaccharides – Different mechanisms for heparin and sulfated galactans. J Biol Chem 2004; 279: 20824-20835.
• 12 Farias WRL, Nazareth RA, Mourão PAS.. Dual effects of sulfated D-galactans from the red algae Botryocladia occidentalis preventing thrombosis and inducing platelet aggregation. Thromb Haemost 2001; 86: 1540-1546.
• 13 Tans G, Rosin J, Griffin JH. Sulfatid-dependent autoactivation of human blood coagulation factor XII (Hageman factor). J Biol Chem 1983; 258: 8215-8222.
• 14 Farndale RW, Buttle DJ, Barret AJ. Improved quantitation and discrimination of sulfated glycosaminoglycans by use of dimethylmethylene blue. Biochim Biophys Acta 1986; 883: 173-177.
• 15 Dubois M, Gilles KA, Hamilton JK. et al. Colorimetric method for determination of sugars and related substances. Anal Chem 1956; 28: 350-356.
• 16 Mourão PAS, Pereira MS, Pavão MSG. 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: 23973-23984.
• 17 Wessler S, Reimer SM, Sheps MC. Biologic assay of thrombosis-inducing activity in human serum. J Appl Physiol 1959; 14: 943-946.
• 18 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.
• 19 Eitzman DT, Westrick RJ, Nabel EG. et al. Plasminogen activator inhibitor-1 and vitronectin promote vascular thrombosis in mice. Blood 2000; 95: 577-580.
• 20 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.
• 21 Fonseca RJC, Mourão PAS.. Fucosylated chondroitin sulfate as a new oral antithrombotic agent. Thromb Haemost 2006; 96: 822-829.
• 22 Herbert JM, Bernat A, Maffrand JP. Importance of platelets in experimental venous thrombosis in the rat. Blood 1992; 80: 2281-2286.
• 23 Mitropoulos KA. High affinity binding of factor XIIa to an electronegative surface controls the rates of factor XII and prekallikrein activation in vitro. Thromb Res 1999; 94: 117-129.
• 24 Kitchen S, Cartwright I, Woods TAL. et al. Lipid composition of seven APTT reagents in relation to heparin sensitivity. Br J Haematol 1999; 106: 801-808.
• 25 Ansell JE. Empowering patients to monitor and manage oral anticoagulation therapy. J Am Med Assoc 1999; 2: 182-183.
• 26 Mousa SA, Linhardt R, Francis JL. et al. Anti-metastatic effect of a non-anticoagulant low-molecularweight heparin versus the standard low-molecularweight heparin, enoxaparin. Thromb Haemost 2006; 96: 816-821.
• 27 Ruf W, Kalnik MW, Lundhansen T. et al. Characterization of factor VII association with tissue factor in solution. J Biol Chem 1991; 266: 15719-15725.
• 28 Renne T, Pozgajova M, Gruner S. et al. Defective thrombus formation in mice lacking coagulation factor XII. J Exp Med 2005; 202: 271-281.
• 29 Dunn JT, Silverberg M, Kaplan AP. The cleavege and formation of activated human Hageman factor by autodigestion and by kallikrein. J Biol Chem 1982; 25: 1779-1784.
• 30 Tans G, Griffin JH. Properties of sulfatides in factor- XII-dependent contact activation. Blood 1982; 59: 69-75.
• 31 Silverberg M, Kaplan AP. Enzymatic activities of activated and zymogen forms of human Hageman Factor (factor XII). Blood 1982; 60: 64-70.
• 32 Daykin HJ, Sturgeon SA, Jones C. et al. Arterial antithrombotic effects of aspirin, heparin, enoxaparin and clopidogrel alone, or in combination, in the rat. Thromb Res 2006; 118: 755-762.