Validated method for rapid inhibition screening of six cytochrome P450 enzymes by liquid chromatography–tandem mass spectrometry

Li, Xiaoyan; Chen, Xiaoyan; Li, Qiang; Wang, Linling; Zhong, Dafang

doi:10.1016/j.jchromb.2007.01.006

Cited by 27 publications

(13 citation statements)

References 25 publications

(40 reference statements)

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“…To expedite these assays, several cocktail approaches, also known as n-in-one assays, have been developed to test for inhibition of several P450 isoforms simultaneously. Most of these assays test for inhibition of five to eight P450 isoforms and use a wide variety of experimental conditions and probe substrates (Dierks et al, 2001;Testino and Patonay, 2003;He et al, 2007;Li et al, 2007;Smith et al, 2007;Workman and Raynaud, 2007;Zientek et al, 2008;Alden et al, 2010;Otten et al, 2011;Yao et al, 2012;Kozakai et al, 2012;Lee and Kim, 2013;Qiao et al, 2014;Qin et al, 2014;Liu et al, 2015). Although a few approaches have claimed using 9 or 10 substrates to evaluate nine isoforms, they actually carry out separate incubations of subsets of probe substrates to avoid P450 interactions before pooling the mixtures immediately prior to a quantitative analysis step, use P450 substrates which are not recommended by the FDA, and/or use long incubation times of up to 60 minutes (Kim et al, 2005;Turpeinen et al, 2005;Tolonen et al, 2007;Dinger et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

High-Throughput Cytochrome P450 Cocktail Inhibition Assay for Assessing Drug-Drug and Drug-Botanical Interactions

Huang

Nikolić

et al. 2015

Drug Metab Dispos

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Detection of drug-drug interactions is essential during the early stages of drug discovery and development, and the understanding of drug-botanical interactions is important for the safe use of botanical dietary supplements. Among the different forms of drug interactions that are known, inhibition of cytochrome P450 (P450) enzymes is the most common cause of drug-drug or drug-botanical interactions. Therefore, a rapid and comprehensive mass spectrometry-based in vitro high-throughput P450 cocktail inhibition assay was developed that uses 10 substrates simultaneously against nine CYP isoforms. Including probe substrates for CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and two probes targeting different binding sites of CYP3A4/5, this cocktail simultaneously assesses at least as many P450 enzymes as previous assays while remaining among the fastest due to short incubation times and rapid analysis using ultrahigh pressure liquid chromatography-tandem mass spectrometry. The method was validated using known inhibitors of each P450 enzyme and then shown to be useful not only for single-compound testing but also for the evaluation of potential drug-botanical interactions using the botanical dietary supplement licorice (Glycyrrhiza glabra) as an example.

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Section: Introductionmentioning

confidence: 99%

High-Throughput Cytochrome P450 Cocktail Inhibition Assay for Assessing Drug-Drug and Drug-Botanical Interactions

Huang

Nikolić

et al. 2015

Drug Metab Dispos

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“…Определение активности различных изоформ цитохромов Р450 осуществляется на основании количества образующегося продукта реакции, при этом для детекции используется в основном масс-спектрометр с тройным квадруполем (QQQ) [74][75][76][77][78][79][80][81][82][83][84][85][86][87]. QQQ состоит из двух квадруполей и находящейся между ними ячейки соударений.…”

Section: методы основанные на вэжх с ультрафиолетовым и масс-спектроunclassified

“…Наличие двух фильтров позволяет получить селективный сигнал исследуемого вещества и повысить чувствительность определения. Анализ литературных данных о наиболее часто используемых субстратах цитохромов Р450 и масс-спектрометрических параметрах, используемых для детектирования суммирован в таблице 3 [74][75][76][77][78][79][80][81][82][83][84][85][86][87].…”

Section: методы основанные на вэжх с ультрафиолетовым и масс-спектроunclassified

“…Например, в статьях [74][75][76][77][78][79][80][81][82] для исследования активности изоформы 1А2 в качестве субстрата использован фенацетин, а его метаболита -ацетоминофен. После инкубации пробы с фенацетином авторы статей [74][75][76][77][78][79][80][81][82] проводили анализ методом обращенно-фазовой жидкостной хроматографии с ионизацией в электроспрее и масс-спектрометрическим детектированием.…”

Section: методы основанные на вэжх с ультрафиолетовым и масс-спектроunclassified

“…После инкубации пробы с фенацетином авторы статей [74][75][76][77][78][79][80][81][82] проводили анализ методом обращенно-фазовой жидкостной хроматографии с ионизацией в электроспрее и масс-спектрометрическим детектированием. Для QQQ в качестве родительского использовали ион [М+Н]+ с m/z 152 Da, соответствующий протонированной форме фенацетина, а в качестве дочернего -фрагмент с m/z 110 Da.…”

Section: методы основанные на вэжх с ультрафиолетовым и масс-спектроunclassified

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Current methods of cytochrome P450 analysis

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BIOMED KHIM

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Current review describes recent approaches of cytochrome P450 concentration and activity evaluation. Special attention paid to modern methods of proteomic analysis such as electrophoresis and chromato-mass-spectrometry. Methods of targeted proteomic applicable for quantitative and qualitative study of P450s in biological samples as well as methods for the enzyme activity measurements are reviewed. Finally, data on correlation between certain P450 isoform content and its specific enzymatic activities were described and discussed in the review.

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In Vitro Liver Systems to Study Induction/Inhibition: Prediction of In Vivo Metabolism and Drug–Drug Interactions

Kono

Iwaki

2012

Encyclopedia of Drug Metabolism and Interactions

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This chapter was the review focused on the evaluation systems for metabolic stability, CYP inhibition and induction not only in the selection process of development candidates including lead optimizations in drug discovery but also to support a design of clinical studies in drug development. In the former process, the advanced in vitro systems have been developed, associating with an inevitable paradigm shift from the throughput to the quality. The similar trend has been gradually transmitted to the prediction of clearance and DDI. The predictions appeared to be in the movement from the basic models to the more mechanistic models including PBPK.

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Validated method for rapid inhibition screening of six cytochrome P450 enzymes by liquid chromatography–tandem mass spectrometry

Cited by 27 publications

References 25 publications

High-Throughput Cytochrome P450 Cocktail Inhibition Assay for Assessing Drug-Drug and Drug-Botanical Interactions

High-Throughput Cytochrome P450 Cocktail Inhibition Assay for Assessing Drug-Drug and Drug-Botanical Interactions

Current methods of cytochrome P450 analysis

In Vitro Liver Systems to Study Induction/Inhibition: Prediction of In Vivo Metabolism and Drug–Drug Interactions

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