title: Is thrombin generation the new rapid, reliable and relevant pharmacological tool for the development of anticoagulant drugs?
abstract: The ex vivo testing emerges as an essential and critical step for the selection of the most promising prospective anticoagulant agents. The aim of the present study was to validate the thrombin generation assay as an ex vivo pharmacological screening test for measuring the anticoagulant behaviour and potency of molecules. The effects of six thrombin and/or factor Xa (FXa) inhibitors (argatroban, lepirudin, PPACK, enoxaparin, ZK-807834, fondaparinux) were investigated on the time course of thrombin catalytic activity triggered by the tissue factor pathway in platelet-poor plasma (PPP) of male healthy volunteers using the Calibrated Automated Thrombogram((R)) (CAT) method. In the presence of the anticoagulant drugs, the thrombin activity profiles were dose-dependently modified according to their specific enzyme inhibitory activity. ZK-807834 was the most potent drug for reducing the C(max) and the V(max) but also for prolonging the T(max). Lepirudin most efficiently delayed the lag time whereas enoxaparin was the most powerfully drug for diminishing the endogenous thrombin potential (ETP). In conclusion, the thrombin activity profile performed with the CAT method is a very rapid, suitable and reliable pharmacological tool for screening thrombin and/or FXa inhibitors whatever their inhibition mode. It consists of a powerful alternative for the classical PT clotting assay, especially regarding to the time course and the total amount of active thrombin generated. Last but not least, it provides insight into the mechanism of action of the compounds.
authors.affiliation: Department of Pharmacy, Drug Design and Discovery Center, FUNDP, University of Namur, Namur, Belgium. severine.robert@fundp.ac.be
authors.forename: Séverine
authors.initials: S
authors.lastname: Robert
authors.forename: Jérémie
authors.initials: J
authors.lastname: Ghiotto
authors.forename: Bernard
authors.initials: B
authors.lastname: Pirotte
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: David
authors.forename: Bernard
authors.initials: B
authors.lastname: Masereel
authors.forename: Lionel
authors.initials: L
authors.lastname: Pochet
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Dogné
mesh_terms: D000925:Anticoagulants / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D006801:Humans; D008297:Male; D013917:Thrombin / Q000037:antagonists & inhibitors* / Q000096:biosynthesis
chemical_list: D000925:Anticoagulants; D013917:Thrombin