RRC ID 57737
著者 Ohashi R, Watanabe R, Esaki T, Taniguchi T, Torimoto-Katori N, Watanabe T, Ogasawara Y, Takahashi T, Tsukimoto M, Mizuguchi K.
タイトル Development of Simplified in Vitro P-Glycoprotein Substrate Assay and in Silico Prediction Models To Evaluate Transport Potential of P-Glycoprotein.
ジャーナル Mol Pharm
Abstract 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/1b 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 data set collected under the same experimental conditions. Most compounds in the test set fell within two- and three-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 that 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.
巻・号 16(5)
ページ 1851-1863
公開日 2019-5-6
DOI 10.1021/acs.molpharmaceut.8b01143
PMID 30933526
MeSH ATP Binding Cassette Transporter, Subfamily B / genetics ATP Binding Cassette Transporter, Subfamily B, Member 1 / metabolism* Animals Biological Availability Cell Membrane Permeability / physiology Central Nervous System Agents / metabolism Computer Simulation* Data Accuracy Drug Discovery / methods* Drug Evaluation, Preclinical / methods* Intestinal Absorption / physiology LLC-PK1 Cells Machine Learning* Protein Transport / physiology* Reproducibility of Results Swine Transfection
IF 4.321
引用数 3
リソース情報
ヒト・動物細胞 LLC-GA5-CoL150(RCB0871)