| KONEČNÝ, Jiří, Jan JUŘICA, Josef TOMANDL and Zdeněk GLATZ. Study of recombinant cytochrome P450 2C9 activity with diclofenac by micellar electrokinetic capillary chromatography (tudy of recombinant cytochrome P450 2C9 activity with diclofenac by micellar electrokinetic capillary chromatography). Electrophoresis. Weinheim: VHC, 2007, vol. 28, No 8, p. 1229-1234. ISSN 0173-0835. |
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@article{699387,
author = {Konečný, Jiří and Juřica, Jan and Tomandl, Josef and Glatz, Zdeněk},
article_location = {Weinheim},
article_number = {8},
keywords = {drug metabolism; cytochrome P450 2C9; diklofenac; MEKC},
language = {eng},
issn = {0173-0835},
journal = {Electrophoresis},
title = {Study of recombinant cytochrome P450 2C9 activity with diclofenac by micellar electrokinetic capillary chromatography},
volume = {28},
year = {2007}
}
TY - JOUR
ID - 699387
AU - Konečný, Jiří - Juřica, Jan - Tomandl, Josef - Glatz, Zdeněk
PY - 2007
TI - Study of recombinant cytochrome P450 2C9 activity with diclofenac by micellar electrokinetic capillary chromatography
JF - Electrophoresis
VL - 28
IS - 8
SP - 1229-1234
EP - 1229-1234
PB - VHC
SN - 01730835
KW - drug metabolism
KW - cytochrome P450 2C9
KW - diklofenac
KW - MEKC
N2 - Cytochrome P450 2C9 (CYP2C9) is one of the most important isoform in human liver involved in the metabolism of a large number of therapeutic agents. The aim of this communication is to demonstrate the applicability of capillary electrophoresis for the determination of the enzymatic activity of CYP2C9 with diclofenac as a probe substrate. Micellar electrokinetic capillary chromatography (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 small quantity of enzymes and substrate. Moreover, the enzymatic reaction can be monitored with high sensitivity and reproducibility even when the reaction mixture is used for the analysis without any pre-treatment. The kinetic study on the given enzymatic reaction was also performed since basic characterization of drug biotranformation generally begins with the enzyme kinetic analysis of metabolite formation. As a result Michaelis constant and maximum reaction velocity were evaluated, the values Km 3.44 uM, respectively 19.78 nmol.min-1.nmol-1 were in agreement with the literature data. On the other hand the slight deviation from typical Michaelis-Menten kinetics with a weak positive cooperativity was found at diclofenac concentration below 2 uM. The same atypical kinetic behaviour of CYP2C9 was also observed by other authors.
ER -
KONEČNÝ, Jiří, Jan JUŘICA, Josef TOMANDL and Zdeněk GLATZ. Study of recombinant cytochrome P450 2C9 activity with diclofenac by micellar electrokinetic capillary chromatography (tudy of recombinant cytochrome P450 2C9 activity with diclofenac by micellar electrokinetic capillary chromatography). \textit{Electrophoresis}. Weinheim: VHC, 2007, vol.~28, No~8, p.~1229-1234. ISSN~0173-0835.
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