Validated assays for human cytochrome P450 activities.

¹Pharacokinetics, Pharmacodynamics, and Drug Metabolism, Pfizer, Inc., Groton, Connecticut 06340, USA.

Drug Metabolism and Disposition: the Biological Fate of Chemicals

|May 25, 2004

Summary

Validated semiautomated assays for human cytochrome P450 (CYP) marker activities ensure reliable in vitro drug interaction data. These methods support confident claims of no in vivo interactions, aiding drug development.

Area of Science:

Pharmacology
Drug Metabolism
Analytical Chemistry

Background:

In vitro drug interaction studies are crucial for drug development.
Accurate in vitro data is essential for predicting in vivo drug interactions.
Validated methods are needed to ensure confidence in in vitro experimental results.

Purpose of the Study:

To develop and validate 12 semiautomated assays for human cytochrome P450 (CYP) marker substrate activities.
To ensure the reliability and accuracy of in vitro drug interaction assessments.
To support the use of in vitro data in lieu of in vivo studies for drug interaction claims.

Main Methods:

Development and validation of 12 semiautomated assays for human CYP marker activities.
Assays validated according to Good Laboratory Practices (GLP).
High-pressure liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) with stable isotope-labeled internal standards used for analysis.

Main Results:

Validated assays cover key human CYP isoforms (CYP1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 3A4, 3A5).
Sensitive HPLC-MS/MS method allowed low microsomal protein concentrations (0.01-0.2 mg/ml).
Excellent assay accuracy and precision; kinetic and inhibition parameters were precise and consistent with literature.

Conclusions:

The validated semiautomated assays provide reliable data for assessing drug interactions.
These methods enhance confidence in using in vitro data to predict in vivo drug interactions.
The assays are suitable for routine assessment of drug candidates' potential for CYP-mediated pharmacokinetic interactions.

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Validated assays for human cytochrome P450 activities.

Area of Science:

Background:

Purpose of the Study:

Main Methods:

Main Results:

Conclusions:

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