並列タイトル等Application of Sandwich-cultured hepatocytes for analysis of drug-drug interaction on bile canalicular transporters
タイトル（掲載誌）平成24(2012)年度 科学研究費補助金 挑戦的萌芽研究 研究成果報告書 = 2012 Fiscal Year Final Research Report
一般注記金沢大学医薬保健研究域薬学系
本研究では肝細胞取り込み能動輸送、細胞内結合、及び肝細胞内生成代謝物の影響を含む薬物間相互作用予測手法の提案を行った。サンドイッチ培養した肝細胞が形成した胆管腔内へのABC輸送体MRP2蛍光プローブ化合物CDFの移行性をたQTLI法により定量化した。その結果CDFの移行性は阻害薬添加時、ならびに生成代謝物が阻害する場合も再現できた。したがって、本手法は、胆管腔側膜輸送体上での、相互作用予測法として有用である。
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.
研究課題/領域番号:23659076, 研究期間(年度):2011-2012
出典：研究課題「胆管側膜輸送体の定量的可視化法の樹立と薬物間相互作用評価系への展開」課題番号23659076（KAKEN：科学研究費助成事業データベース（国立情報学研究所））（https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-23659076/23659076seika/）を加工して作成
一次資料へのリンクURLhttps://kanazawa-u.repo.nii.ac.jp/?action=repository_action_common_download&item_id=43120&item_no=1&attribute_id=26&file_no=1
関連情報https://kaken.nii.ac.jp/search/?qm=20155237
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23659076/
https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-23659076/23659076seika/
連携機関・データベース国立情報学研究所 : 学術機関リポジトリデータベース（IRDB）（機関リポジトリ）