Comparison of in vitro P-glycoprotein screening assays: recommendations for their use in drug discovery.
The ATP-dependent drug efflux pump P-glycoprotein (P-gp) affects the absorption and disposition of many compounds. P-gp may also play role in clinically significant drug-drug interactions. Therefore, it is important to find potential substrates or inhibitors of P-gp early in the drug discovery process. To identify compounds that interact with this ... transporter, several P-gp assays were validated and compared by testing a set of 28 reference compounds, including inhibitors of cytochrome P450 3A4 (CYP3A4). The assays included in silico predictions, inhibition assays (based on cellular uptake of rhodamine-123 or calcein AM), and functional assays (ATPase activity assay and transcellular transport assay, the latter for a subset of compounds). In addition, species differences were studied in an indirect fluorescence indicator screening assay and test systems expressing porcine, mouse, or human P-gp. Our results suggest that several P-gp assays should be used in combination to classify compounds as substrates or inhibitors of P-gp. Recommendations are given on screening strategies which can be applied to different phases of the drug discovery and development process.
Mesh Terms:
Adenosine Triphosphatases, Animals, Biological Transport, Cells, Cultured, Drug Evaluation, Preclinical, Fluoresceins, Fluorescent Dyes, Humans, Indicators and Reagents, Mice, Models, Molecular, P-Glycoprotein, Rhodamines, Species Specificity, Swine
Adenosine Triphosphatases, Animals, Biological Transport, Cells, Cultured, Drug Evaluation, Preclinical, Fluoresceins, Fluorescent Dyes, Humans, Indicators and Reagents, Mice, Models, Molecular, P-Glycoprotein, Rhodamines, Species Specificity, Swine
J. Med. Chem.
Date: Apr. 24, 2003
PubMed ID: 12699389
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