Preparation, purification and common characteristics of anticoagulant peptide from peloid

Abstract

Aim. To obtain the peloid extract containing peptide anticoagulant, to separate it from co-extracted compounds, to confirm the chemical structure of the purified anticoagulant, to characterize the amino acid profile and to evaluate its effect on blood clotting activity in vitro and the effect on hemostasis in laboratory animals. Methods. The anticoagulant was derived from lake Taraskul (Tyumen region) peloid. The effector was prepared and purified by original method including extraction, sedimentation of humic acids by salting-out, and removal of fulvic acids and low molecular weight fractions of humic acids by ion exchange chromatography. Purity and homogeneity of gained anticoagulant was evaluated by high performance liquid chromatography and polyacrylamide gel electrophoresis. The chemical composition of the active substance was determined by acid hydrolysis and identification of phenylthiohydantoin amino acid derivatives by reversed-phase high performance liquid chromatography. Specific anticoagulant activity was evaluated by substance’s influence on effect of thrombin-dependent conversion of fibrinogen to fibrin and serum thrombin clotting time after being administered to laboratory animals. Results. Peptide anticoagulant was isolated of the peloid and purified. Its activity is realized at the stage of thrombin and fibrinogen interaction. The substance prolongs the fibrin clotting time both in a model test system and when administered intravenously to rats. The anticoagulant effect of the peptide when administered to animals lasts at least 60 min, and no significant toxic effects appear. Conclusion. An isolated active substance might be a basis for the drug development of anticoagulants.