Oral administration of dermatan sulphate reduces venous thrombus formation in vivo: potential use as a formulation for venous thromboembolism.

Dermatan sulphate (DS) is a sulphated polysaccharide that displays complexity in constituent sulphated disaccharides and interacts with proteins and signalling molecules to modulate numerous biological processes, including inhibition of the coagulation cascade and regulation of blood clotting and fibrinolysis. This study shows the antithrombotic and anticoagulant effects of DS prepared from bovine collagen waste liquor following oral and intravenous administrations in a deep vein thrombosis (DVT) rabbit model. In vitro, the prothrombin time, activated partial thromboplastin time, and thrombin citrated plasma clotting assays revealed that bovine DS had strong antithrombotic and anticoagulant effects comparable to low-molecular-weight heparin [Clexane® (enoxaparin sodium)]. In a DVT rabbit model, animals received intravenous and oral administrations of bovine DS and Clexane® providing further evidence that both agents had strong antithrombotic and anticoagulant effects by significantly reducing or preventing clot formation. Thromboelastography (TEG) assays revealed further that both bovine DS and Clexane® substantially prolonged the clotting time of recalcified citrated whole blood, but only bovine DS could retain clot strength suggesting that bovine DS had less effect on platelet-fibrin interactions. In conclusion, this is the first report that oral administration of DS from bovine collagen waste liquor reduces experimental venous thrombus formation warranting further research into bovine DS as an oral antithrombotic therapeutic.

Effect of glycomacropeptide fractions on cholecystokinin and food intake.

Glycomacropeptide (GMP) is the hydrophilic 64-amino acid C-terminal glycopeptide released into cheese whey when kappa-casein is cleaved by chymosin. GMP exists as a mixture of different glycoforms due to the carbohydrates sialic acid (N-acetylneuraminic acid, NeuNAc), galactose (Gal), galactosamine and glucosamine attached by O-glycosidic linkages. GMP reportedly stimulates the release of cholecystokinin (CCK), which may promote satiety. The objectives of the present study were to manufacture three glycoforms of GMP, minimally glycosylated GMP (3.5 (sd 0.1) % NeuNAc and 1.5 (sd 0.1) % Gal), glycosylated GMP (12.0 (sd 0.3) % NeuNAc and 4.2 (sd 0.2) % Gal) and a GMP-depleted whey protein concentrate, and to assess the effects of these fractions relative to glucose on CCK, subjective measures of satiety and food intake. In a randomised double-blind acute study, twenty overweight/obese males (56.9 (sd 7.2) years, 97.4 (sd 8.1) kg, 31.5 (sd 3.0) kg/m2) were recruited to consume four 50 g preloads (two GMP preparations, GMP-depleted whey and glucose) containing 895 kJ. Blood samples and subjective measures of satiety were collected before and at 15, 30, 60, 90, 120 and 180 min after the consumption of preload, and CCK levels were measured. A lunchtime meal of hot food was provided from which subjects ate ad libitum until satisfied. Energy and nutrient intakes from the food consumed were calculated. There was no significant difference in CCK levels, subjective measures of satiety or food intake between treatments at the given preload level. These results suggest that the protein fractions at the dose employed do not influence satiety, CCK levels or energy intake at a subsequent meal.

Antithrombin activity and disaccharide composition of dermatan sulfate from different bovine tissues.

Dermatan sulfate is a glycosaminoglycan that selectively inhibits the action of thrombin through interaction with heparin cofactor II. Unlike heparin it does not interact with other coagulation factors and is able to inhibit thrombin associated with clots. This property has made dermatan sulfate an attractive candidate as an antithrombotic drug. Previous studies have showed that dermatan sulfate derived from porcine/bovine intestinal mucosa/skin or marine invertebrates is capable of stimulating heparin cofactor II-mediated thrombin inhibition in vitro. This biological activity is reported for the first time in this study using dermatan sulfate derived from mammalian tissues other than intestinal mucosa or skin. Ten different bovine tissues including the aorta, diaphragm, eyes, large and small intestine, esophagus, skin, tendon, tongue, and tongue skin were used to prepare dermatan sulfate-enriched fractions by anion exchange chromatography and acetone precipitation. Heparin cofactor II/dermatan sulfate-mediated thrombin inhibition measured in vitro revealed activity comparable to or higher than the commercial standard with 2-fold differences observed between some tissues. Analysis of the extracted dermatan sulfate using fluorophore-assisted carbohydrate electrophoresis revealed significant differences in the relative percentage of all the mono-sulfated disaccharides, in particular the predominant mammalian disaccharide uronic acid-->N-acetyl-D-galactosamine-4-O-sulfate, confirming previous reports regarding variations in sulfation in dermatan sulfate from different tissues. Overall, these findings demonstrate that dermatan sulfate extracted from a range of bovine tissues exhibits in vitro antithrombin activity equivalent to or higher than that observed for porcine intestinal mucosa, identifying additional sources of dermatan sulfate as potential antithrombotic agents.