2007
DOI: 10.1002/elps.200600560
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Study of recombinant cytochrome P450 2C9 activity with diclofenac by MEKC

Abstract: Cytochrome P450 2C9 (CYP2C9) is one of the most important isoforms in human liver involved in the metabolism of a large number of therapeutic agents. The aim of this paper is to demonstrate the applicability of CE for the determination of the enzymatic activity of CYP2C9 with diclofenac as a probe substrate. MEKC with SDS as a pseudostationary phase was used for this purpose. Compared to other assays, the MEKC-based method is rapid, can be automated and requires only a small quantity of enzymes and substrate. … Show more

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Cited by 19 publications
(13 citation statements)
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“…. The obtained values of K m ′ = 2.66 ± 0.18 μM, V max ′ = 7.91 ± 0.22 nmol min −1 nmol ‑1 and n = 1.59 ± 0.16 are in a good agreement with literature data determined by other techniques .…”
Section: Resultssupporting
confidence: 90%
See 1 more Smart Citation
“…. The obtained values of K m ′ = 2.66 ± 0.18 μM, V max ′ = 7.91 ± 0.22 nmol min −1 nmol ‑1 and n = 1.59 ± 0.16 are in a good agreement with literature data determined by other techniques .…”
Section: Resultssupporting
confidence: 90%
“…On the basis of the statistical evaluation of the obtained data , 50 mM phosphate (pH 7.4) was chosen as the incubation buffer. Compared to the initial MEKC conditions , the ionic strength of the sample was considerably lower and it was therefore possible to separate the reaction products by means of CZE. A BGE composed of 20 mM sodium dihydrogen phosphate, 20 mM disodium tetraborate buffer (pH 8.6), and the application of 23 kV (356.6 V/cm, positive polarity) were selected as appropriate separation conditions on the basis of the migration time and resolution of the analyte peak.…”
Section: Resultsmentioning
confidence: 99%
“…As was mentioned above, the main aim of this study was to confirm the atypical (non‐Michaelis–Menten) kinetic behaviour of CYP2C9 with diclofenac as the probe substrate at low concentrations. Since this concentration lies close to the LOQ of the previously developed MEKC method 8, it was first necessary to find appropriate conditions for more sensitive determination of reaction product – 4′‐hydroxydiclofenac. Several approaches can be used to solve this problem in CE.…”
Section: Resultsmentioning
confidence: 99%
“…A MEKC method has recently been developed for the kinetic study of the reaction of the cytochrome P450 2C9 isoform (CYP2C9) with diclofenac as a probe substrate 8. Although CYP2C9, one of the most important isoforms in the human liver, constitutes about 20% of total human liver CYP and metabolizes approximately 10% of therapeutically important drugs 9, 10, diclofenac is a widely used nonsteroidal anti‐inflammatory drug which acts as a potent inhibitor of prostaglandin endoperoxide synthase 11.…”
Section: Introductionmentioning
confidence: 99%
“…Metabolism of flurbiprofene by CYP2C9 in vitro was significantly activated by dapsone in microsomes, but only a minor effect was observed in vivo (Hutzler et al, 2001a). Homotropic cooperativity (n H = 1.4) of diclofenac hydroxylation by CYP2C9 was observed in (Konecny et al, 2007). Heterotropic effects in CYP2C9 have been reviewed (Egnell et al, 2003; Tracy & Hummel, 2004; Tracy, 2006), also see Section 7.…”
Section: Homotropic Cooperativity In Steady-state Kinetics Of Subsmentioning
confidence: 98%