2013
New Capillary Electrophoretic Method for On-line Screenings of Drug Metabolism Mediated by Cytochrome P450 Enzymes
ŘEMÍNEK, Roman, Marta ZEISBERGEROVÁ, Monika LANGMAJEROVÁ a Zdeněk GLATZZákladní údaje
Originální název
New Capillary Electrophoretic Method for On-line Screenings of Drug Metabolism Mediated by Cytochrome P450 Enzymes
Název česky
Nová metoda pro on-line studie metabolismu léčiv pomocí CE
Autoři
ŘEMÍNEK, Roman (203 Česká republika, garant, domácí), Marta ZEISBERGEROVÁ (203 Česká republika, domácí), Monika LANGMAJEROVÁ (203 Česká republika) a Zdeněk GLATZ (203 Česká republika, domácí)
Vydání
Electrophoresis, Weinheim, WILEY-VCH Verlag GmbH, 2013, 0173-0835
Další údaje
Jazyk
angličtina
Typ výsledku
Článek v odborném periodiku
Obor
10600 1.6 Biological sciences
Stát vydavatele
Německo
Utajení
není předmětem státního či obchodního tajemství
Impakt faktor
Impact factor: 3.161
Kód RIV
RIV/00216224:14310/13:00066068
Organizační jednotka
Přírodovědecká fakulta
UT WoS
000327590900013
Klíčová slova česky
CE; Cytochrom P450; Metabolismus léčiv;
Klíčová slova anglicky
CE; Cytochrome P450; Drug Metabolism;
Příznaky
Mezinárodní význam, Recenzováno
Změněno: 25. 4. 2014 14:59, Ing. Zdeňka Rašková
V originále
A new method for the determination of kinetic and inhibition parameters of cytochromes P450 reactions by means of on-line capillary electrophoresis was developed. It is based on transverse diffusion of laminar flow profiles methodology introduced by Krylov et al. which injection procedure was modified. The solutions of an enzyme and its substrates are injected by hydrodynamic pressure as a series of repeated consecutive plugs. Proposed injection of 3 plugs of enzyme surrounded with plugs of substrates represents a certain trade-off to obtain the reaction mixture with the satisfying homogeneity by the short injection procedure as possible. Mathematical modelling confirmed the assumption of a consistent distribution of reactants in the final reaction mixture. Kinetic and inhibition studies of cytochrome P450 2C9’s reaction with diclofenac as a probe substrate and sulfaphenazole as a probe inhibitor were conducted in order to prove the practical applicability of the proposed method for on-line screenings of drug metabolism mediated by cytochrome P450 enzymes. As a result, an apparent Michaelis constant of 2.66 +- 0.18 uM, apparent maximum reaction velocity of 7.91 +- 0.22 nmol min-1 nmol-1, Hill coefficient of 1.59 +- 0.16, half maximal inhibitory concentration of 0.94 +- 0.04 uM and apparent inhibition constant of 0.39 +- 0.07 uM were determined. All these values are in agreement with literature data obtained using different techniques. In addition, less than 30 nL of cytochrome P450 2C9 solution was consumed per analysis in the kinetic and inhibition studies using this method.
Česky
Byla nová metoda pro on-line studie metabolismu léčiv pomocí CE
Návaznosti
EE2.3.20.0182, projekt VaV |
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GAP206/10/0057, projekt VaV |
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