Structure-activity relationship of the pro- and anticoagulant effects of Fucus vesiculosus fucoidan

Summary

Fucoidan is a highly complex sulfated polysaccharide commonly extracted from brown seaweed. In addition to their many biological activities, fucoidans have recently been demonstrated to inhibit or increase coagulation at different concentration ranges. Their structural features, i.e. molecular weight (Mw), Mw distribution, degree of sulfation, monosaccharide composition, and different linkages, are known to affect these activities. Therefore, structure-activity relationship (SAR) analysis of fucoidan is crucial for its potential use as a procoagulant. In this study, Fucus vesiculosus (F.v.) fucoidan was fractionated by charge and size as well as over- and desulfated to different degrees to yield preparations with various structural properties. The fractions’ pro- and anticoagulant activities were assessed by calibrated automated thrombography (CAT) and activated partial thromboplastin time (aPTT) assays. Binding to and inhibition of the anticoagulant protein tissue factor pathway inhibitor (TFPI) and the ability to activate coagulation via the contact pathway were also investigated. This paper discusses the impact of charge density, size, and sugar composition on fucoidan’s pro- and anticoagulant activities. Fucoidan requires a minimal charge density of 0.5 sulfates per sugar unit and a size of 70 sugar units to demonstrate desired procoagulant activities for improvement of haemostasis in factor VIII/factor IX-deficient plasma.

Keywords

Blood coagulation - fucoidan - molecular weight - structure-activity relationship - sulfates

^* These authors contributed equally.

• References

• 1 Berteau O, Mulloy B. Sulfated fucans, fresh perspectives: structures, functions, and biological properties of sulfated fucans and an overview of enzymes active toward this class of polysaccharide. Glycobiology 2003; 13: 29R-40R.
• 2 Pomin VH, Mourao PA. Structure, biology, evolution, and medical importance of sulfated fucans and galactans. Glycobiology 2008; 18: 1016-1027.
• 3 Liu T, Scallan CD, Broze Jr. GJ. et al. Improved coagulation in bleeding disorders by Non-Anticoagulant Sulfated Polysaccharides (NASP). Thromb Hae-most 2006; 95: 68-76.
• 4 Prasad S, Lillicrap D, Labelle A. et al. Efficacy and safety of a new-class hemos-tatic drug candidate, AV513, in dogs with hemophilia A. Blood 2008; 111: 672-679.
• 5 Hilden I, Lauritzen B, Sorensen BB. et al. Hemostatic effect of a monoclonal antibody mAb 2021 blocking the interaction between FXa and TFPI in a rabbit hemophilia model. Blood 2012; 119: 5871-5878.
• 6 Waters EK, Genga RM, Schwartz MC. et al. Aptamer ARC19499 mediates a pro-coagulant hemostatic effect by inhibiting tissue factor pathway inhibitor. Blood 2011; 117: 5514-5522.
• 7 Dockal M, Hartmann R, Polakowski T. et al. Improvement of hemostasis in hemophilia by peptides inhibiting tissue factor pathway inhibitor. J Thromb Hae-most 2011; 9: 1-250.
• 8 Skriptsova A, Shevchenko N, Zvyagintseva T. et al. Monthly changes in the content and monosaccharide composition of fucoidan from Undaria pinnatifida (Laminariales, Phaeophyta). J Appl Phycol 2010; 22: 79-86.
• 9 Zhang Z, Li B, Suwan J. et al. Analysis of pharmaceutical heparins and potential contaminants using (1)H-NMR and PAGE. J Pharm Sci 2009; 98: 4017-4026.
• 10 Farias WR, 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.
• 11 Fonseca RJ, Oliveira SN, Melo FR. et al. Slight differences in sulfation of algal galactans account for differences in their anticoagulant and venous antithrom-botic activities. Thromb Haemost 2008; 99: 539-545.
• 12 Pereira MS, Mulloy B, Mourao PA. 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.
• 13 Pereira MS, Melo FR, Mourao PA. Is there a correlation between structure and anticoagulant action of sulfated galactans and sulfated fucans?. Glycobiology 2002; 12: 573-580.
• 14 Ustyuzhanina NE, Ushakova NA, Zyuzina KA. et al. Influence of fucoidans on hemostatic system. Mar Drugs 2013; 11: 2444-2458.
• 15 Handa N. Examination on the Applicability of the Phenol Sulfuric Acid Method to the Determination of Dissolved Carbohydrate in Sea Water. J Oceanograph Soc Japan 1966; 22: 79-86.
• 16 Qiu X, Amarasekara A, Doctor V. Effect of oversulfation on the chemical and biological properties of fucoidan. Carbohydrate Polymers 2006; 63: 224-228.
• 17 Miller IJ, Blunt JW. Desulfation of algal galactans. Carbohydrate Res 1998; 309: 39-43.
• 18 Zhang Z, Khan NM, Nunez KM. et al. Complete monosaccharide analysis by high-performance anion-exchange chromatography with pulsed amperometric detection. Anal Chem 2012; 84: 4104-4110.
• 19 Hemker HC, Giesen P, al Dieri R. et al. Calibrated automated thrombin generation measurement in clotting plasma. Pathophysiol Haemost Thromb 2003; 33: 4-15.
• 20 Knappe S, Gorczyca ME, Jilma B. et al. Plasmatic tissue factor pathway inhibitor is a major determinant of clotting in factor VIII inhibited plasma or blood. Thromb Haemost 2013; 109: 450-457.
• 21 Melo FR, Mourao PA. An algal sulfated galactan has an unusual dual effect on venous thrombosis due to activation of factor XII and inhibition of the coagulation proteases. Thromb Haemost 2008; 99: 531-538.
• 22 Muller F, Mutch NJ, Schenk WA. et al. Platelet polyphosphates are proinflam-matory and procoagulant mediators in vivo. Cell 2009; 139: 1143-1156.
• 23 Guerrini M, Beccati D, Shriver Z. et al. Oversulfated chondroitin sulfate is a contaminant in heparin associated with adverse clinical events. Nat Biotechnol 2008; 26: 669-675.
• 24 Kishimoto TK, Viswanathan K, Ganguly T. et al. Contaminated heparin associated with adverse clinical events and activation of the contact system. N Engl J Med 2008; 358: 2457-2467.
• 25 Dargaud Y, Luddington R, Baglin TP. Elimination of contact factor activation improves measurement of platelet-dependent thrombin generation by calibrated automated thrombography at low-concentration tissue factor. J Thromb Haemost 2006; 4: 1160-1161.
• 26 Broze Jr. GJ, Girard TJ. Tissue factor pathway inhibitor: structure-function. Front Biosci 2012; 17: 262-280.
• 27 Li B, Lu F, Wei X. et al. Fucoidan: structure and bioactivity. Molecules 2008; 13: 1671-1695.
• 28 Haroun-Bouhedja F, Ellouali M, Sinquin C. et al. Relationship between sulfate groups and biological activities of fucans. Thromb Res 2000; 100: 453-459.
• 29 Chen S, Li G, Wu N. et al. Sulfation pattern of the fucose branch is important for the anticoagulant and antithrombotic activities of fucosylated chondroitin sulfates. Biochim Biophys Acta 2013; 1830: 3054-3066.