Improvement of the chemical inhibition phenotyping assay by cross-reactivity correction

Nicholas M. Njuguna; Ken-ichi Umehara; Felix Huth; Hilmar Schiller; Kelly Chibale; Gian Camenisch

doi:10.1515/dmpt-2016-0028

Article

Improvement of the chemical inhibition phenotyping assay by cross-reactivity correction

Nicholas M. Njuguna , Ken-ichi Umehara , Felix Huth , Hilmar Schiller , Kelly Chibale and Gian Camenisch

Published/Copyright: October 8, 2016

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From the journal Drug Metabolism and Personalized Therapy Volume 31 Issue 4

Abstract

Background:

The fraction of an absorbed drug metabolized by the different hepatic cytochrome P450 (CYP) enzymes, relative to total hepatic CYP metabolism (fm_CYP), can be estimated by measuring the inhibitory effects of presumably selective CYP inhibitors on the intrinsic metabolic clearance of a drug using human liver microsomes. However, the chemical inhibition data are often affected by cross-reactivities of the chemical inhibitors used in this assay.

Methods:

To overcome this drawback, the cross-reactivities exhibited by six chemical inhibitors (furafylline, montelukast, sulfaphenazole, ticlopidine, quinidine and ketoconazole) were quantified using specific CYP enzyme marker reactions. The determined cross-reactivities were used to correct the in vitrofm_CYPs of nine marketed drugs. The corrected values were compared with reference data obtained by physiologically based pharmacokinetics simulation using the software SimCYP.

Results:

Uncorrected in vitrofm_CYPs of the nine drugs showed poor linear correlation with their reference data (R²=0.443). Correction by factoring in inhibitor cross-reactivities significantly improved the correlation (R²=0.736).

Conclusions:

Correcting in vitro chemical inhibition results for cross-reactivities appear to offer a straightforward and easily adoptable approach to provide improved fm_CYP data for a drug.

Keywords: chemical inhibition; cross-reactivity; CYP; enzyme phenotyping; fmCYP; physiologically based pharmacokinetics (PBPK) model

Acknowledgments

The study was conducted with the great support by the Next Generation Scientist program, which was organized by the Diversity and Inclusion Office of Novartis Pharma AG. The University of Cape Town, South African Medical Research Council, and South African Research Chairs Initiative of the Department of Science and Technology administered through the South African National Research Foundation are gratefully acknowledged for support. The authors wish to acknowledge Dr. Markus Zollinger for the critical evaluation of this work. Special thanks go to Bertrand-Luc Birlinger, Claire Juif, Judith Streckfuss and Marc Witschi for their support in the data generation.

Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: Declaration or None declared.
Employment or leadership: Declaration or None declared.
Honorarium: Declaration or None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Received: 2016-8-29

Accepted: 2016-9-13

Published Online: 2016-10-8

Published in Print: 2016-12-1

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Articles in the same Issue

https://doi.org/10.1515/dmpt-2016-0028

Keywords for this article

chemical inhibition; cross-reactivity; CYP; enzyme phenotyping; fmCYP; physiologically based pharmacokinetics (PBPK) model