Assessment of Transporter-Mediated and Passive Hepatic Uptake Clearance Using Rifamycin-SV as a Pan-Inhibitor of Active Uptake
- 11 July 2018
- journal article
- research article
- Published by American Chemical Society (ACS) in Molecular Pharmaceutics
- Vol. 15 (10), 4677-4688
- https://doi.org/10.1021/acs.molpharmaceut.8b00654
Abstract
The use of in vitro data for the quantitative prediction of transporter-mediated clearance is critical. Central to this evaluation is the use of hepatocytes since they contain the full complement of transporters and metabolic enzymes. In general, uptake clearance (CLuptake) is evaluated by measuring appearance of compound in the cell. Passive clearance (CLpd) is often determined by conducting parallel studies at 4°C or by attempting to saturate uptake pathways. Both approaches have their limitations. Recent studies have proposed the use Rifamycin-SV (RFV) as a pan-inhibitor of hepatic uptake pathways. In our studies, we confirm that transport activity of all major hepatic uptake transporters is inhibited significantly by RFV 1 mM (OATP1B1, 1B3 and 2B1 = NTCP (80%), OCT1 (65%), OAT2 (60%)). Under these incubation conditions, we found that the free intracellular concentration of RFV is ~175 µM and that several major CYPs and UGTs can be reversibly inhibited. Using this approach, we also determined CLuptake and CLpd of nine known OATP substrates across three different lots of human hepatocytes. The scaling factors generated for these compounds at 37 °C with RFV and 4 °C were found to be similar. Interestingly, CLpd of passively permeable compounds like metoprolol and semagacestat were found to be higher at 37 °C compared to 4 °C indicating a temperature effect on these compounds. In addition, our data also suggests that incorporation of medium concentrations into CLuptake and CLpd calculations may be critical for highly protein bound and highly lipophilic drugs. Overall our data indicate that RFV, instead of 4 °C, can be reliably used to measure CLuptake and CLpd of drugs.Keywords
This publication has 44 references indexed in Scilit:
- Prediction of Pharmacokinetics and Drug–Drug Interactions When Hepatic Transporters are InvolvedClinical Pharmacokinetics, 2014
- The role of drug metabolizing enzymes in clearanceExpert Opinion on Drug Metabolism & Toxicology, 2014
- A Perspective on the Prediction of Drug Pharmacokinetics and Disposition in Drug Research and DevelopmentDrug Metabolism and Disposition, 2013
- Clinical significance of organic anion transporting polypeptides (OATPs) in drug disposition: their roles in hepatic clearance and intestinal absorptionBiopharmaceutics & Drug Disposition, 2013
- Species differences in drug transporters and implications for translating preclinical findings to humansExpert Opinion on Drug Metabolism & Toxicology, 2012
- Applications of Human Pharmacokinetic Prediction in First-in-Human Dose EstimationThe AAPS Journal, 2012
- Prediction of Human Drug Clearance by Multiple Metabolic Pathways: Integration of Hepatic and Intestinal Microsomal and Cytosolic DataDrug Metabolism and Disposition, 2011
- Membrane transporters in drug developmentNature Reviews Drug Discovery, 2010
- P-glycoprotein Expression, Localization, and Function in Sandwich-Cultured Primary Rat and Human Hepatocytes: Relevance to the Hepatobiliary Disposition of a Model Opioid PeptidePharmaceutical Research, 2004
- Organ Clearance Concepts: New Perspectives on Old PrinciplesJournal of Pharmacokinetics and Biopharmaceutics, 1997