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Home / Drug Metabolism Letters, Volume 5, Number 2

Evaluation of Different Approaches to Identifying a Higher Throughput Assay for Time-Dependent Inhibition (TDI)

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Authors: Kajbaf, M.; Longhi, R.; Fontana, S.

Source: Drug Metabolism Letters, Volume 5, Number 2, 2011, pp. 104-113(10)

Publisher: Bentham Science Publishers

DOI: https://doi.org/10.2174/187231211795305230

The availability of a reliable in vitro assay to evaluate time-dependent inhibition (TDI) of cytchrome P450 enzymes by novel compounds is essential for the identification of candidate medicines. We have evaluated three assay methods, making use of 59 marketed compounds and 28 novel GSK compounds. Recombinant bactosomes expressing the CYP3A4 isozyme were used with two fluorescence-based methods: a “Re-addition” assay and a “30 min” assay. The third method evaluated used pooled human liver microsomes (PHLM) with LC-MS/MS detection (the data for GSK compounds were evaluated in this study, whereas data for marketed drugs were reported recently). Our evaluation showed that the Re-addition method is comparable to the LC-MS/MS method in terms of predictivity and reproducibility. In conclusion, Re-addition method is inexpensive, and provides a simple assessment of the risk of TDI for novel compounds. This assay is particularly appropriate for use during the early stages of drug discovery.





Keywords: CYP2D6; Diethoxyfluoresin (DEF); Time-dependent inhibition (TDI); cerivastatin; diethoxyfluorescein (DEF); mibefradil; midazolam; pooled human liver microsomes; recombinant CYP3A4 isozyme

Document Type: Research Article

Publication date: 01 April 2011

More about this publication?
  • Drug Metabolism Letters publishes short papers on major advances in all areas of drug metabolism and disposition. The emphasis will be on publishing quality papers very rapidly. Letters will be processed rapidly by taking full advantage of the Internet technology for both the submission and review of manuscripts. The journal covers the following areas:

    In vitro systems including CYP-450; enzyme induction and inhibition; drug-drug interactions and enzyme kinetics; pharmacokinetics, toxicokinetics, species scaling and extrapolations; P-glycoprotein and transport carriers; target organ toxicity and interindividual variability; drug metabolism and disposition studies; extrahepatic metabolism; phase I and phase II metabolism; recent developments for the identification of drug metabolites, reactive intermediate and glutathione conjugates.
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