High‐throughput cytochrome P450 (CYP) inhibition screening via a cassette probe‐dosing strategy. VI. Simultaneous evaluation of inhibition potential of drugs on human hepatic isozymes CYP2A6, 3A4, 2C9, 2D6 and 2E1

Bu, Hai‐Zhi; Magis, Lisa; Knuth, Kim; Teitelbaùm, Philip

doi:10.1002/rcm.290

Cited by 75 publications

(57 citation statements)

References 15 publications

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“…Although strategies have been developed to increase throughput by techniques such as sample pooling, multiple assay compression, high-speed LC/MS (Bu et al, 2001;Zhang et al, 2002), single-concentration IC 50 projection (Gao et al, 2002), and virtual screening (Rao et al, 2000;Zuegge et al, 2002), the analytical approach still remains a challenge that limits sample throughput.…”

mentioning

confidence: 99%

In Vitro Drug Interactions of Cytochrome P450: An Evaluation of Fluorogenic to Conventional Substrates

Cohen¹,

Remley²,

Raunig³

et al. 2003

Drug Metab Dispos

124

109

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This article is available online at http://dmd.aspetjournals.org ABSTRACT:Clinically observed drug interactions with cytochrome P450 (P450) enzymes have increased the need to assess drug interactions of new chemical entities early in the discovery process. To meet this need, fluorogenic substrates have been commercialized. However, only limited evaluations of their utility and comparisons to drug probes have been reported. This study examines the correlation between IC 50 values obtained with fluorogenic and conventional drug probes for structurally diverse inhibitors of the five major human P450 isoforms. In general, correlations are weak, with significant numbers of compounds being missed as inhibitors by either probe. For P450s 1A2, 2C9, and 2C19, correlation coefficients were above 0.6 with slopes that ranged from 1.5 to 4.2.However, for P450s 1A2 and 2C9, about 20% of compounds were not included because an IC 50 value could not be determined with one of the two probes. CYP 2C19 had the highest correlation (correlation coefficient 0.84), with a slope of 2.0 and less than 5% of compounds excluded. CYP 2D6 showed a good correlation for IC 50 values less than 10 M. However, at higher IC 50 values, a high degree of scatter was observed. CYP 3A4 had the weakest correlation, and a large number of compounds were excluded with the fluorogenic probe. Overall, the study shows the care needed when selecting fluorogenic probes and the caution needed when results with fluorogenic probes are used to drive structure-activity relationships with respect to drug interactions.

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mentioning

confidence: 99%

In Vitro Drug Interactions of Cytochrome P450: An Evaluation of Fluorogenic to Conventional Substrates

Cohen¹,

Remley²,

Raunig³

et al. 2003

Drug Metab Dispos

124

109

View full text Add to dashboard Cite

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“…Currently, the most widely employed incubation sample preparation is protein precipitation (PPT) [11][12][13][14][15][16][17][18][19]. Our initial approach for sample preparation was based on PPT, while it was found that liquid-liquid extraction provided a 10-fold higher sensitivity for the majority of the CYP probe substrate metabolites than PPT.…”

Section: Sample Preparationmentioning

confidence: 99%

“…At present, numerous high-throughput approaches using fluorogenic substrates to measure CYP activities have been described [8,9], as well as substrate "cocktail" experiments that simultaneously measure more than one CYP activity [10][11][12][13][14][15][16][17][18]. Recently Kim et al [19] described a high-throughput method that allowed the simultaneous evaluation of the metabolic activities of nine major human CYP isoforms (i.e., CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4) on their well-known specific probe substrates using human hepatic microsomes.…”

Section: Introductionmentioning

confidence: 99%

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

Chen

et al. 2007

Journal of Chromatography B

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“…In order to estimate the metabolism in the human body, human liver microsomal preparations are often used to evaluate the in vitro metabolism of the lead compound. 168,169 LC/MS/MS using a triple quadrupole instrument is an appropriate and powerful means for that purpose. The full-scan mode can easily provide information on changes in the molecular weight due to metabolism; in addition, the neutral loss scan and precursor scan, which are unique to the MS/MS instrument, can also easily identify the metabolites in most contaminants.…”

Section: ·2 Drug Metabolismmentioning

confidence: 99%

Biomedical and Biological Mass Spectrometry

Mano

Goto

2003

ANAL. SCI.

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By the end of the 20th century, mass spectrometry (MS) had evolved drastically as a major analytical tool in biomedical and biological research. Recent mass spectrometers, designed for ease of handling, came into wide use in the 1990s. Soft ionization methods, which were made practical in the second half of the 1980s, allowed for the sensitive analysis of nonvolatile and thermolabile compounds by MS. Electrospray ionization (ESI) 1,2 and matrix-assisted laser desorption ionization (MALDI) 3 methods, made it possible to ionize large biomolecules and to transfer them to the gas phase, thereby becoming a valuable tool in both protein and peptide analysis [4][5][6] and in the study of drugs and their metabolites [7][8][9][10][11][12] in the human body. Many reviews of MS have recently been published, [13][14][15][16][17][18][19] and in particular, the number of reviews describing biochemical, biological, and biomedical MS have increased dramatically over the last ten years. Initially, there were many commentary reviews on new techniques, such as soft ionization methods and mass analyzers, 13-15 and more recently, review articles have focused on the biological and biomedical applications of MS. 18-21 Above all, two major fields, drug discovery and proteomics, have made marked advances based on MS.Articles in the MEDLINE database were searched, and the results are illustrated in Fig. 1 and Table 1. Over the last 15 years, a total of 79220 MS articles were placed in the database, with a 2.5-fold increase from 1987 to 1999. The number of articles using fast atom bombardment (FAB) as an ionization method remained almost constant during this period. This ionization method is a practical soft ionization procedure, which was applied to the analysis of peptides [22][23][24][25][26][27][28][29][30][31] and endogenous biological substances [32][33][34][35] in the 1980s. Atmospheric pressure chemical ionization (APCI) is a suitable method 36 for the ionization of middle or weakly polar low-molecular-weight molecules, and can be easily coupled to conventional highperformance liquid chromatography (HPLC), used at a flow rate of around 1 mL/min. [37][38][39][40][41][42][43][44][45][46][47][48] However, the number of articles concerning APCI have increased slowly, because most drugs developed recently contain polar groups which can be ionized without difficulty by the ESI technique. Japan This review focuses on biological and biomedical mass spectrometry, and covers a selection of publications in this area included in the MEDLINE database for the period 1987 -2001. Over the last 15 years, biological and biomedical mass spectrometry has progressed out of all recognition. The development of soft ionization methods, such as electrospray ionization and matrix-assisted laser desorption ionization, has mainly contributed to the remarkable progress, because they can easily produce gas-phase ions of large, polar, and thermally labile biomolecules, such as proteins, peptides, nucleic acids and others. The innovations of ionization meth...

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High‐throughput cytochrome P450 (CYP) inhibition screening via a cassette probe‐dosing strategy. VI. Simultaneous evaluation of inhibition potential of drugs on human hepatic isozymes CYP2A6, 3A4, 2C9, 2D6 and 2E1

Cited by 75 publications

References 15 publications

In Vitro Drug Interactions of Cytochrome P450: An Evaluation of Fluorogenic to Conventional Substrates

In Vitro Drug Interactions of Cytochrome P450: An Evaluation of Fluorogenic to Conventional Substrates

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

Biomedical and Biological Mass Spectrometry

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