title: 3,6-disubstituted coumarins as mechanism-based inhibitors of thrombin and factor Xa.
abstract: In this work, coumarins were screened on thrombin (THR) and factor Xa (FXa), two of the most promising targets for the development of anticoagulant drugs. This allowed us to highlight compound 30, characterized by a 2,5-dichlorophenyl ester in the 3-position and a chloromethyl moiety in the 6-position, as a very potent THR inhibitor (ki/KI= 37,000 M(-1) s(-1)). Moreover, this compound exhibits good selectivity over FXa (168-fold) and trypsin (54-fold). The mechanism of inactivation was investigated in this series and significantly differs from that previously observed with alpha-chymotrypsin. Indeed, the addition of hydrazine on the THR-inhibitor complex promotes a partial induced THR reactivation. This reactivation, confirmed by LC/MS, showed the resurgence of the native THR and a new dihydrazide complex. Docking experiments were then efficiently used to explain the trends observed in the enzymatic assays as well as to corroborate the postulated inhibition mechanism. Finally, the cell permeability of our derivatives was estimated using a computational approach.
authors.affiliation: Department of Pharmacy and CBS Laboratory, University of Namur, FUNDP, 61, Rue de Bruxelles, B-5000 Namur, Belgium.
authors.forename: Raphaël
authors.initials: R
authors.lastname: Frédérick
authors.forename: Séverine
authors.initials: S
authors.lastname: Robert
authors.forename: Caroline
authors.initials: C
authors.lastname: Charlier
authors.forename: Jérôme
authors.initials: J
authors.lastname: de Ruyck
authors.forename: Johan
authors.initials: J
authors.lastname: Wouters
authors.forename: Bernard
authors.initials: B
authors.lastname: Pirotte
authors.forename: Bernard
authors.initials: B
authors.lastname: Masereel
authors.forename: Lionel
authors.initials: L
authors.lastname: Pochet
mesh_terms: D000818:Animals; D018938:Caco-2 Cells; D002417:Cattle; D002463:Cell Membrane Permeability; D002853:Chromatography, Liquid; D003198:Computer Simulation; D003374:Coumarins / Q000138:chemical synthesis* / Q000737:chemistry; D004789:Enzyme Activation; D015951:Factor Xa / Q000737:chemistry; D065427:Factor Xa Inhibitors; D006801:Humans; D007700:Kinetics; D013058:Mass Spectrometry; D008958:Models, Molecular; D013329:Structure-Activity Relationship; D013917:Thrombin / Q000037:antagonists & inhibitors* / Q000737:chemistry
chemical_list: C507717:6-chloromethyl-3-(2,5-dichlorophenoxycarbonyl)-2-benzopyrone; D003374:Coumarins; D065427:Factor Xa Inhibitors; D013917:Thrombin; D015951:Factor Xa