2012 Fiscal Year Final Research Report
Application of Sandwich-cultured hepatocytes for analysis of drug-drug interaction on bile canalicular transporters
| Project/Area Number |
23659076
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Medical pharmacy
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| Research Institution | Kanazawa University |
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Principal Investigator |
TAMAI Ikumi 金沢大学, 薬学系, 教授 (20155237)
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| Co-Investigator(Kenkyū-buntansha) |
NAKANISHI Takeo 金沢大学, 薬学系, 准教授 (30541742)
KUNISHIMA Munetaka 金沢大学, 薬学系, 教授 (10214975)
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| Project Period (FY) |
2011 – 2012
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| Keywords | 輸送体 / 肝細胞 / 薬物間相互作用 / 副作用 / 胆汁排泄 / 可視化 |
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| Research Abstract |
Interplay of transporters and enzymes is essential to understand drug disposition in tissues such as liver and intestine. When we consider drug-drug interaction (DDI) on liver bile canalicular transporters,metabolites formed in the hepatocytes must be considered, since many of conjugated metabolites show higher affinity to those transporters like MRP2. We established a quantitative time-lapse imaging-based analysis (QTLI) to assess Mrp2-mediated DDIs in sandwich-cultured hepatocytes (SCHs), utilizing fluorescent probe substrate of MRP2, (5,6)-carboxy-2’,7’-dichlorofluorescein (CDF). When estradiol (E2) was chosen as affecting compound, its metabolite estradiol-17β-glucuronide (E17G) but not E2 itself was confirmed to inhibit Mrp2-mediated CDF transport. When SCRHs were preincubated with E2, fluorescence accumulated in bile canaliculi formed in SCRH was decreased depending upon both length of preincubation period and concentration of E2 given in extracellular medium. The decrease in accumulated fluorescence agreed with an increase in intracellular concentration of E17G generated in hepatocytes, suggesting that the phase II biotransformation is mirrored in MRP2-mediated transport by QTLI. Since SCHs well maintain hepatic uptake transport activity, intracellular binding and drug metabolizing activity as in vitro system, QTLI in SCHs provides a convenient platform to develop an evaluation system for transporter-based DDIs without identifying metabolites of drug candidates.
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