Characterization of human warfarin reductase
Doctoral thesis, Master thesis
mesheuropmc: cardiovascular diseases | heterocyclic compounds
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Simona Sokolová Supervisor: PharmDr. Petra Malátková, Ph.D. Title of diploma thesis: Characterization of human warfarin reductase Warfarin is widely used anticoagulant drug. Considering the narrow therapeutic window of warfarin, it is important to fully understand its metabolism in human body. Oxidative, reductive and conjugation reactions are involved in warfarin metabolism. However, the reductive metabolism of warfarin has not been studied in details until now. The reduced metabolite of warfarin, i.e. warfarin-alcohol, is produced by the conversion of the carbonyl group of the side chain. It is known that human liver cytosolic and microsomal fractions exhibit warfarin reductase activity but the specific enzymes catalysing the reduction of warfarin are not known yet. The aim of this study was to identify the enzyme(s) participating in reduction of warfarin and to describe enzyme kinetics. Human liver cytosolic and microsomal fractions and recombinant enzymes AKR1A1, AKR1B1, AKR1B10, AKR1C1, AKR1C2, AKR1C3, AKR1C4, CBR1 and CBR3 were incubated with warfarin at various concentrations. The produced warfarin-alcohol was quantified by UHPLC and the specific activities of enzymes and...