Luminogenic cytochrome P450 assays

被引：154

作者：

Cali, James J. ^{[1
]}

Ma, Dongping ^{[1
]}

Sobol, Mary ^{[1
]}

Simpson, Daniel J. ^{[1
]}

Frackman, Susan ^{[1
]}

Good, Troy I. ^{[1
]}

Daily, William J. ^{[1
]}

Liu, David ^{[1
]}

机构：

[1] Promega Corp, Madison, WI 53711 USA

来源：

EXPERT OPINION ON DRUG METABOLISM & TOXICOLOGY | 2006年 / 2卷 / 04期

关键词：

CYP; CYP induction; CYP inhibition; cytochrome P450; drug-drug interaction; drug metabolism; fluorescent P450 assay; hepatocyte; high-throughput screening; liver microsomes; luminescent P450 assay; P450-Glo (TM); recombinant CYP enzymes;

D O I：

10.1517/17425255.2.4.629

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Luminogenic cytochrome P450 (CYP) assays couple CYP enzyme activity to firefly luciferase luminescence in a technology called P450-Glo (TM) (Promega). Luminogenic substrates are used in assays of human CYP1A1, -1A2, -1B1, -2C8, -2C9, -2C19, -2D6, -2J2, -3A4, -3A7; -4A11, -4F3B, -4F12 and -19. The assays detect dose-dependent CYP inhibition by test compounds against recombinant CYP enzymes or liver microsomes. Induction or inhibition of CYP activities in cultured hepatocytes is measured in a nonlytic approach that leaves cells intact for additional analysis: Luminogenic CYP assays offer advantages of speed and safety over HPLC and radiochemical-based methods. Compared with fluorogenic methods the approach offers advantages of improved sensitivity and decreased interference between optical properties of test compound and CYP substrate. These homogenous assays are sensitive and robust tools for high-throughput CYP screening in early drug discovery.

引用

页码：629 / 645

页数：17

共 76 条

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