Bednarczyk, Dallas (2021) Passive influx and ion trapping are more relevant to the cellular accumulation of highly permeable low molecular weight acidic drugs than is Organic Anion Transporter 2 (OAT2). Drug metabolism and disposition: the biological fate of chemicals. ISSN 1521009X

Abstract

Recently published work suggests that highly permeable low molecular weight (LMW) acidic drugs are transported by Organic Anion Transporter 2 (OAT2). However, an asymmetric distribution of ionizable drugs in subcellular organelles where pH gradients are significant may occur in the presence of an inhibitor relative to its absence (e.g. lysosomal trapping). In the present study, OAT2-mediated transport of highly permeable LMW anions could not be demonstrated using OAT2 transfected cells, despite robust transport of the OAT2 substrate penciclovir. Moreover, a rifamycin SV (RifSV) dependent reduction in the accumulation of highly permeable LMW anions previously observed in hepatocytes could be qualitatively reproduced using HepG2 cells and also in MDCK cells which lack expression of OAT2. Neither HepG2 nor MDCK cells demonstrated meaningful penciclovir transport, nor was the cellular accumulation of the highly permeable LMW anions sensitive to competitive inhibition by the neutral OAT2 substrate penciclovir. Both cell lines however demonstrated sensitivity to the mitochondrial uncoupler p-trifluoromethoxy carbonyl cyanide phenyl hydrazone (FCCP) in a manner similar to RifSV. Furthermore, the transepithelial MDCK permeability of the highly permeable LMW anions was measured in the absence and presence of RifSV and FCCP at concentrations that reduced the cellular accumulation of anions. Neither inhibitor, nor the OAT2 inhibitor ketoprofen, reduced the transepithelial flux of the anions as would be anticipated for transported substrate inhibition. The findings presented here are aligned with cellular accumulation of highly permeable LMW anions being significantly determined by ion trapping sensitive to mitochondrial uncoupling rather than the result of OAT2-mediated transport. Significance Statement The manuscript illustrates that passive influx and ion trapping are more relevant to the cellular accumulation of highly permeable low molecular weight acidic drugs than is the previously proposed mechanism of OAT2-mediated transport. The outcome illustrated here highlights a rare, and perhaps previously not reported, observation of anionic drug trapping in a compartment sensitive to mitochondrial uncoupling (e.g. the mitochondrial matrix) that may be confused for transporter-mediated uptake.

Item Type:	Article
Keywords:	mitochondria organic anion transport Organic anion uptake / efflux (OATs, OATPs)
Date Deposited:	06 Jul 2021 00:45
Last Modified:	06 Jul 2021 00:45
URI:	https://oak.novartis.com/id/eprint/44349