Generic Method for Cytochrome P450 Metabolism Studies Employing Short End Injection Capillary Electrophoresis Mode
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| Year of publication | 2011 |
| Type | Conference abstract |
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| Description | Cytochromes P450 (CYP) play a key role in metabolism of more than 60 % currently prescribed drugs [1], therefore stability against this group of enzymes represents one of the important drug parameters. Pharmaceutical companies strive to determine the action of CYP on a new drug candidate early within development process in order to eliminate compounds with inappropriate qualities and thus economize following tests. Human liver microsomes (HLM) constitute a favourable screening media for this purpose offering fast analyses, high CYP concentration and good simulation of metabolism inside the liver tissue. The aim of this study was to introduce a method based on capillary electrophoresis (CE) allowing both assessment of drug stability against HLM and determination of main CYP isoform responsible for a metabolism of a given drug. The method based on NADP+ production monitoring was employed to ensure its generic character enabling screening of large groups of miscellaneous compounds. Conceptually, particular CYP isoform responsible for metabolism of a new drug candidate is then determined by decrease in NADP+ production in reaction with its specific inhibitor. Finally, the generic method enabling fast assessment of relation between a new drug candidate and CYP in human liver was established. Separation of reaction mixture components was achieved within 2 minutes analysis with 20 mM phosphate-borate buffer pH 8.6 primarily due to combination of CE short end injection mode and application of high voltage of -30 kV. Presented method thus represents a promising tool for high-throughput screenings of candidate compounds within early stages of a new drug development. |
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