3,6-disubstituted coumarins as mechanism-based inhibitors of thrombin and factor Xa.
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.
Frédérick
Raphaël
R
Department of Pharmacy and CBS Laboratory, University of Namur, FUNDP, 61, Rue de Bruxelles, B-5000 Namur, Belgium.
Robert
Séverine
S
Charlier
Caroline
C
de Ruyck
Jérôme
J
Wouters
Johan
J
Pirotte
Bernard
B
Masereel
Bernard
B
Pochet
Lionel
L
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
C507717:6-chloromethyl-3-(2,5-dichlorophenoxycarbonyl)-2-benzopyrone; D003374:Coumarins; D065427:Factor Xa Inhibitors; D013917:Thrombin; D015951:Factor Xa