Development of a Simplified in Vitro P-glycoprotein Substrate Assay and in Silico Prediction Models to Evaluate Transport Potential of P-glycoprotein.

08:00 EDT 1st April 2019 | BioPortfolio

Summary of "Development of a Simplified in Vitro P-glycoprotein Substrate Assay and in Silico Prediction Models to Evaluate Transport Potential of P-glycoprotein."

For efficient drug discovery and screening, it is necessary to simplify P-glycoprotein (P-gp) substrate assays and to provide in silico models that predict the transport potential of P-gp. In this study, we developed a simplified in vitro screening method to evaluate P-gp substrates by unidirectional membrane transport in P-gp-overexpressing cells. The unidirectional flux ratio positively correlated with parameters of the conventional bidirectional P-gp substrate assay (R2 = 0.941) and in vivo Kp, brain ratio (mdr1a/b KO/WT) in mice (R2 = 0.800). Our in vitro P-gp substrate assay had high reproducibility and required approximately half the labor of the conventional method. We also constructed regression models to predict the value of P-gp-mediated flux and three-class classification models to predict P-gp substrate potential (low-, medium-, and high-potential) using 2397 data entries with the largest dataset collected under the same experimental conditions. Most compounds in the test set fell within 2- and 3-fold errors in the random forest regression model (71.3 and 88.5%, respectively). Furthermore, the random forest three-class classification model showed a high balanced accuracy of 0.821 and precision of 0.761 for the low-potential classes in the test set. We concluded that the simplified in vitro P-gp substrate assay was suitable for compound screening in the early stages of drug discovery and the in silico regression model and three-class classification model using only chemical structure information could identify the transport potential of compounds including P-gp-mediated flux ratios. Our proposed method is expected to be a practical tool to optimize effective central nervous system (CNS) drugs, to avoid CNS side effects, and to improve intestinal absorption.


Journal Details

This article was published in the following journal.

Name: Molecular pharmaceutics
ISSN: 1543-8392


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