New Capillary Electrophoretic Method for On-line Screenings of
Drug Metabolism Mediated by Cytochrome P450 Enzymes

J 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 GLATZ

Základní 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

DOI

http://dx.doi.org/10.1002/elps.201300124

UT WoS

000327590900013

Klíčová slova česky

CE; Cytochrom P450; Metabolismus léčiv;

Klíčová slova anglicky

CE; Cytochrome P450; Drug Metabolism;

Štítky

AKR, rivok, ZR

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 25. 4. 2014 14:59, Ing. Zdeňka Rašková

Anotace

ORIG CZ

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

Název: Vytvoření multidisciplinárního výzkumného a vzdělávacího centra bioanalytických technologií

GAP206/10/0057, projekt VaV

Název: Miniaturizovaný systém pro "on-line" studie metabolismu léčiv založený na kapilární elektroforéze

Investor: Grantová agentura ČR, Miniaturizovaný systém pro "on-line" studie metabolismu léčiv založený na kapilární elektroforéze

Citovat

ŘEMÍNEK, Roman, Marta ZEISBERGEROVÁ, Monika LANGMAJEROVÁ a Zdeněk GLATZ. New Capillary Electrophoretic Method for On-line Screenings of Drug Metabolism Mediated by Cytochrome P450 Enzymes. Electrophoresis. Weinheim: WILEY-VCH Verlag GmbH, 2013, roč. 34, č. 18, s. 2705-2711. ISSN 0173-0835. Dostupné z: https://dx.doi.org/10.1002/elps.201300124.

@article{1093552,
   author = {Řemínek, Roman and Zeisbergerová, Marta and Langmajerová, Monika and Glatz, Zdeněk},
   article_location = {Weinheim},
   article_number = {18},
   doi = {http://dx.doi.org/10.1002/elps.201300124},
   keywords = {CE; Cytochrome P450; Drug Metabolism;},
   language = {eng},
   issn = {0173-0835},
   journal = {Electrophoresis},
   title = {New Capillary Electrophoretic Method for On-line Screenings of Drug Metabolism Mediated by Cytochrome P450 Enzymes},
   volume = {34},
   year = {2013}
}

TY  - JOUR
ID  - 1093552
AU  - Řemínek, Roman - Zeisbergerová, Marta - Langmajerová, Monika - Glatz, Zdeněk
PY  - 2013
TI  - New Capillary Electrophoretic Method for On-line Screenings of Drug Metabolism Mediated by Cytochrome P450 Enzymes
JF  - Electrophoresis
VL  - 34
IS  - 18
SP  - 2705-2711
EP  - 2705-2711
PB  - WILEY-VCH Verlag GmbH
SN  - 01730835
KW  - CE
KW  - Cytochrome P450
KW  - Drug Metabolism;
N2  - 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.
ER  -

ŘEMÍNEK, Roman, Marta ZEISBERGEROVÁ, Monika LANGMAJEROVÁ a Zdeněk GLATZ. New Capillary Electrophoretic Method for On-line Screenings of Drug Metabolism Mediated by Cytochrome P450 Enzymes. \textit{Electrophoresis}. Weinheim: WILEY-VCH Verlag GmbH, 2013, roč.~34, č.~18, s.~2705-2711. ISSN~0173-0835. Dostupné z: https://dx.doi.org/10.1002/elps.201300124.

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