Quantitative Contribution of Six Major Transporters to the Hepatic Uptake of Drugs: “SLC-Phenotyping” Using Primary Human Hepatocytes
Open Access
- 11 April 2019
- journal article
- research article
- Published by American Society for Pharmacology & Experimental Therapeutics (ASPET) in Journal of Pharmacology and Experimental Therapeutics
- Vol. 370 (1), 72-83
- https://doi.org/10.1124/jpet.119.257600
Abstract
Hepatic uptake transporters (solute carriers, SLCs), including organic anion transporting polypeptide (OATP)1B1, OATP1B3, OATP2B1, sodium-dependent taurocholate co-transporting polypeptide (NTCP), organic anion (OAT2) and organic cation (OCT1) transporters, play a key role in determining the systemic and liver exposure of chemically diverse drugs. Here, we established a phenotyping approach to quantify the contribution of the six SLCs, and passive diffusion, to the overall uptake using plated human hepatocytes (PHH). First, selective inhibitor conditions were identified by screening about 20 inhibitors across the six SLCs, using single-transfected HEK293 cells. Data implied rifamycin SV (20 µM) inhibits three OATPs, while rifampicin (5µM) inhibits OATP1B1/1B3 only. Further, hepatitis B virus myristoylated-preS1 peptide (HBVpep, 0.1 µM), quinidine (100 µM) and ketoprofen (100-300 µM) are relatively selective against NTCP, OCT1 and OAT2, respectively. Second, using these inhibitory conditions, the fraction transported (ft) by the individual SLCs was characterized for 20 substrates with PHH. Generally, extended clearance classification system (ECCS) class 1A/3A (e.g., warfarin) and 1B/3B compounds (e.g., statins) showed predominant OAT2 and OATP1B1/1B3 contribution, respectively. OCT1-mediated uptake was prominent for class 2/4 compounds (e.g., metformin). Third, in vitro ft values were corrected using quantitative proteomics data to obtained 'scaled ft'. Fourth, in vitro-in vivo extrapolation of the scaled OATP1B1/1B3 ft was assessed leveraging statin clinical drug-drug interaction data with rifampicin as perpetrator. Finally, we outlined a novel step-wise strategy to implement phenotypic characterization of SLC-mediated hepatic uptake for new molecular entities and drugs in a drug discovery and development setting.Keywords
This publication has 53 references indexed in Scilit:
- Dissecting the relative contribution of OATP1B1-mediated uptake of xenobiotics into human hepatocytes using siRNAXenobiotica, 2013
- Clinical significance of organic anion transporting polypeptides (OATPs) in drug disposition: their roles in hepatic clearance and intestinal absorptionBiopharmaceutics & Drug Disposition, 2013
- Classification of Inhibitors of Hepatic Organic Anion Transporting Polypeptides (OATPs): Influence of Protein Expression on Drug–Drug InteractionsJournal of Medicinal Chemistry, 2012
- Determination of OATP-, NTCP- and OCT-mediated substrate uptake activities in individual and pooled batches of cryopreserved human hepatocytesEuropean Journal of Pharmaceutical Sciences, 2011
- Further Characterization of the Electrogenicity and pH Sensitivity of the Human Organic Anion-Transporting Polypeptides OATP1B1 and OATP1B3Molecular Pharmacology, 2010
- Membrane transporters in drug developmentNature Reviews Drug Discovery, 2010
- Design, Data Analysis, and Simulation of in Vitro Drug Transport Kinetic Experiments Using a Mechanistic in Vitro ModelDrug Metabolism and Disposition, 2008
- Ritonavir, Saquinavir, and Efavirenz, but Not Nevirapine, Inhibit Bile Acid Transport in Human and Rat HepatocytesThe Journal of pharmacology and experimental therapeutics, 2006
- PREDOMINANT CONTRIBUTION OF ORGANIC ANION TRANSPORTING POLYPEPTIDE OATP-B (OATP2B1) TO APICAL UPTAKE OF ESTRONE-3-SULFATE BY HUMAN INTESTINAL CACO-2 CELLSDrug Metabolism and Disposition, 2006
- The Intestinal First-pass Metabolism of Substrates of CYP3A4 and P-glycoprotei—Quantitative Analysis Based on Information from the LiteratureDrug Metabolism and Pharmacokinetics, 2003