title: Application of quantitative time-lapse imaging (QTLI) for evaluation of Mrp2-based drug-drug interaction induced by liver metabolites.
abstract: We previously reported a quantitative time-lapse imaging (QTLI)-based analysis method to assess drug-drug interactions (DDI) at multidrug resistance-associated protein 2 (Mrp2) in rat sandwich-cultured hepatocyte (SCH) system, utilizing the fluorescent Mrp2 substrate, 5-(and 6)-carboxy-2',7'-dichlorofluorescein (CDF). Here, we aimed to examine the feasibility of using QTLI to evaluate DDI involving drug metabolite(s) generated in hepatocytes. We used estradiol (E2) and bilirubin as model compounds; both are not substrates of MRP2, whereas their hepatic metabolites, estradiol-17β-glucuronide (E17G) or bilirubin glucuronides, are known to be its substrates as well as inhibitors. When rat SCHs were pre-exposed with E2, fluorescence of CDF accumulated in bile canaliculi decreased depending upon both the duration of pre-exposure and the concentration of extracellular E2. The decrease corresponded with the increase in intracellular concentration of E17G in hepatocytes. Furthermore, cytotoxicity of vinblastine, a substrate of MRP2, was enhanced in SCHs treated with E2. Similarly, CDF accumulated in bile canaliculi was significantly reduced in rat SCHs pre-exposed with bilirubin. In conclusion, these results suggest that phase II biotransformation of a competitor is reflected in alteration of MRP2-mediated CDF transport detected in QTLI. The QTLI might provide a convenient platform to evaluate transporter-based DDIs involving hepatic metabolites of drug candidates without the need to identify the metabolites.
authors: Department of Membrane Transport and Biopharmaceutics, Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa 920‐1192, Japan.
authors: Takeo
authors: T
authors: Nakanishi
authors: Miho
authors: M
authors: Ikenaga
authors: Hajime
authors: H
authors: Fukuda
authors: Norikazu
authors: N
authors: Matsunaga
authors: Ikumi
authors: I
authors: Tamai
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D001663:Bilirubin / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D018929:Cell Culture Techniques; D004347:Drug Interactions; D004958:Estradiol / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D005240:Feasibility Studies; D005452:Fluoresceins / Q000737:chemistry; D022781:Hepatocytes / Q000378:metabolism*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D059008:Time-Lapse Imaging / Q000379:methods*; D014747:Vinblastine / Q000494:pharmacology*
chemical_list: D018528:ATP-Binding Cassette Transporters; C423013:Abcc2 protein, rat; D000972:Antineoplastic Agents, Phytogenic; D005452:Fluoresceins; C053897:5(6)-carboxy-2',7'-dichlorofluorescein; C025483:estradiol-17 beta-glucuronide; C016266:bilirubin glucuronate; D004958:Estradiol; D014747:Vinblastine; D001663:Bilirubin