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Capillary electrophoretic investigation of the enantioselective metabolism of propafenone by human cytochrome P‐450 SUPERSOMES: Evidence for atypical kinetics by CYP2D6 and CYP3A4
Author(s) -
Afshar Minoo,
Thormann Wolfgang
Publication year - 2006
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200500664
Subject(s) - chemistry , stereoselectivity , enantiomer , propafenone , hydroxylation , cyp3a4 , stereochemistry , enantioselective synthesis , enzyme kinetics , cytochrome p450 , kinetics , enzyme , cyp2d6 , biochemistry , catalysis , active site , cardiology , medicine , physics , quantum mechanics , atrial fibrillation
An enantioselective CE method was used to identify the ability of CYP450 enzymes and their stereoselectivity in catalyzing the transformation of propafenone (PPF) to 5‐hydroxy‐propafenone (5OH‐PPF) and N ‐despropyl‐propafenone (NOR‐PPF). Using in vitro incubations with single CYP450 enzymes (SUPERSOMES), 5OH‐PPF is shown to be selectively produced by CYP2D6 and N ‐dealkylation is demonstrated to be mediated by CYP2D6, CYP3A4, CYP1A2, and CYP1A1. For the elucidation of kinetic aspects of the metabolism with CYP2D6 and CYP3A4, incubations with individual PPF enantiomers and racemic PPF were investigated. With the exception of the dealkylation in presence of R ‐PPF only, which can be described by the Michaelis–Menten model, all CYP2D6‐induced reactions were found to follow autoactivation kinetics. For CYP3A4, all NOR‐PPF enantiomer formation rates as function of PPF enantiomer concentration were determined to follow substrate inhibition kinetics. The formation of NOR‐PPF by the different enzymes is stereoselective and is reduced significantly when racemic PPF is incubated. Clearance values obtained for CYP3A4 dealkylation are stereoselective whereas those of CYP2D6 hydroxylation are not. This paper reports the first investigation of the PPF hydroxylation and dealkylation kinetics by the CYP2D6 enzyme and represents the first report in which enantioselective CE data provide the complete in vitro kinetics of metabolic steps of a drug.