title: Substrate and method dependent inhibition of three ABC-transporters (MDR1, BCRP, and MRP2).
abstract: Drug transport and drug-drug interactions (DDI) with human ABC transporters are generally investigated in mammalian cell lines or inverted membrane vesicles from insect cells (Sf9) overexpressing the transporter of interest. In this study, we instead used membrane vesicles from human embryonic kidney cells (HEK293) overexpressing wild type MDR1/Pgp (ABCB1), BCRP (ABCG2), and MRP2 (ABCC2) with the aim to study the concentration dependent inhibition of shared and prototypic probe substrates. We first investigated 15 substrates and identified estrone-17-beta-glucorinide (E17G) as shared substrate. Nine specific and general inhibitors were then studied using E17G and prototypic probe substrates. The results were compared with those previously obtained in Sf9 vesicles and cell lines of canine (MDCKII) and human (Saos-2) origin. For the majority of inhibitors, Kivalues differed <10-fold between E17G and probe substrates. Significant differences in Kivalues were observed for about one third of the inhibitors. The transport inhibition potencies in HEK293 vesicles were in good agreement with those obtained in Sf9 vesicles. Large differences were found in the inhibition potencies observed in the vesicular systems compared to the cellular systems. Nevertheless, the rank order correlations between the different experimental systems were generally good. Our study provides further information on substrate dependent inhibition of ABC-transporters, and suggests that simple ranking of compounds can be used as a tier one approach to bridge results obtained in different experimental systems.
authors.affiliation: Department of Pharmacy, Uppsala University, Uppsala, Sweden. Electronic address: jenny.pedersen@qps.com.
authors.forename: Jenny M
authors.initials: JM
authors.lastname: Pedersen
authors.affiliation: Department of Pharmacy, Uppsala Univeristy, Uppsala, Sweden.
authors.forename: Elin K
authors.initials: EK
authors.lastname: Khan
authors.affiliation: Department of Pharmacy, Uppsala Univeristy, Uppsala, Sweden.
authors.forename: Christel A S
authors.initials: CAS
authors.lastname: Bergström
authors.affiliation: Pharmaceutical Technology and Development, AstraZeneca R&D Gothenburg, SE-43183 Mölndal, Sweden.
authors.forename: Johan
authors.initials: J
authors.lastname: Palm
authors.affiliation: CNS and Pain Innovative Medicines DMPK, AstraZeneca R&D, Södertälje, Sweden.
authors.forename: Janet
authors.initials: J
authors.lastname: Hoogstraate
authors.affiliation: Department of Pharmacy and Uppsala University Drug Optimization and Pharmaceutical Profiling Platform, Chemical Biology Consortium, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.
authors.forename: Per
authors.initials: P
authors.lastname: Artursson
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B / Q000037:antagonists & inhibitors; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2 / Q000037:antagonists & inhibitors*; D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D004285:Dogs; D004970:Estrone / Q000031:analogs & derivatives* / Q000737:chemistry; D020719:Glucuronides / Q000737:chemistry*; D006801:Humans; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000037:antagonists & inhibitors*; D061987:Sf9 Cells; D013329:Structure-Activity Relationship
chemical_list: D018435:ATP Binding Cassette Transporter, Subfamily B; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D004970:Estrone; D020719:Glucuronides; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C513055:ABCB1 protein, human; C000715087:ABCC2 protein, human; C000625731:estrone-17-glucuronide
keywords: ATP binding cassette transporter; DDI, drug-drug-interaction; HEK293; Membrane vesicles; Transport inhibition; Transport protein