J 2013

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

ŘEMÍNEK, Roman, Marta ZEISBERGEROVÁ, Monika LANGMAJEROVÁ and Zdeněk GLATZ

Basic information

Original name

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

Name in Czech

Nová metoda pro on-line studie metabolismu léčiv pomocí CE

Authors

ŘEMÍNEK, Roman (203 Czech Republic, guarantor, belonging to the institution), Marta ZEISBERGEROVÁ (203 Czech Republic, belonging to the institution), Monika LANGMAJEROVÁ (203 Czech Republic) and Zdeněk GLATZ (203 Czech Republic, belonging to the institution)

Edition

Electrophoresis, Weinheim, WILEY-VCH Verlag GmbH, 2013, 0173-0835

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Germany

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 3.161

RIV identification code

RIV/00216224:14310/13:00066068

Organization unit

Faculty of Science

DOI

UT WoS

000327590900013

Keywords (in Czech)

CE; Cytochrom P450; Metabolismus léčiv;

Keywords in English

CE; Cytochrome P450; Drug Metabolism;

Tags

Tags

International impact, Reviewed
Změněno: 25/4/2014 14:59, Ing. Zdeňka Rašková

Abstract

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.

In Czech

Byla nová metoda pro on-line studie metabolismu léčiv pomocí CE

Links

EE2.3.20.0182, research and development project
Name: Vytvoření multidisciplinárního výzkumného a vzdělávacího centra bioanalytických technologií
GAP206/10/0057, research and development project
Name: Miniaturizovaný systém pro "on-line" studie metabolismu léčiv založený na kapilární elektroforéze
Investor: Czech Science Foundation, Miniaturized on-line drug metabolism system based on capillary electrophoresis