Fluorescence-based assays for the assessment of drug interaction with the human transporters OATP1B1 and OATP1B3.
Fluorescence-based assays for the assessment of drug interaction with the human transporters OATP1B1 and OATP1B3.
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DOI：
10.1016/j.ab.2010.06.012
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发表时间：
2010-10-01
影响因子：
2.9
通讯作者：
Bednarczyk D
中科院分区：
文献类型：
--
作者：
Bednarczyk D
肝倾向在多种药物的药代动力学和药效学中起着重要作用。窦膜转运体已被证明参与许多药物的肝脏处置。OATP1B1和OATP1B3是两种已确定在肝脏配置中起作用的窦膜转运蛋白。OATP1B1和OATP1B3与他汀类药物的肝脏摄取有关，与OATP1B1相关的多态性与有害的患者终点有关。因此，OATP1B1和OATP1B3代表了潜在的药物-药物相互作用位点。有许多方法可以识别潜在的药物-药物与转运体的相互作用。然而，相对较少提供基于荧光的检测的便利性和速度。在这里，我们开发了一种基于荧光的检测方法，用于测量OATP1B1和OATP1B3介导的8-荧光素- camp （8-FcA）的转运。OATP1B1和OATP1B3介导的8-FcA转运具有时间依赖性和可饱和性（Km = 2.9 μM, Vmax = 0.20 pmol/min/cm2和1.8 μM和0.33 pmol/min/cm2）。已知与oatp相互作用的分子，包括环孢素A、利福平和格列本脲，均显示出OATP1B1和OATP1B3对8-FcA转运的浓度依赖性抑制。本文描述的以8-FcA为底物的体外荧光检测方法方便、快速，并且在筛选候选药物与OATP1B1和OATP1B3的潜在药物相互作用方面具有实用价值。
Hepatic disposition plays a significant role in the pharmacokinetics and pharmacodynamics of a variety drugs. Sinusoidal membrane transporters have been shown to participate in the hepatic disposition of many pharmaceuticals. Two sinusoidal membrane transporters with an established role in hepatic disposition are OATP1B1 and OATP1B3. OATP1B1 and OATP1B3 have been implicated in the hepatic uptake of statin drugs and polymorphisms linked to OATP1B1 have been associated with deleterious patient endpoints. As a result, OATP1B1 and OATP1B3 represent sites for potential drug-drug interactions. Numerous methods exist for identifying potential drug-drug interactions with transporters. However, relatively few offer the convenience and speed of fluorescence-based assays. Here, a fluorescence-based assay was developed for measuring the OATP1B1 and OATP1B3 mediated transport of 8-fluorescein-cAMP (8-FcA). The OATP1B1 and OATP1B3 mediated transport of 8-FcA was time dependent and saturable (Km = 2.9 μM, Vmax = 0.20 pmol/min/cm2 and 1.8 μM and 0.33 pmol/min/cm2, respectively). Molecules known to interact with OATPs, including cyclosporin A, rifampicin, and glibenclamide, each demonstrated concentration dependent inhibition of 8-FcA transport by OATP1B1 and OATP1B3. The in vitro fluorescence-based assays described here using 8-FcA as the substrate are convenient, rapid, and have utility in screening drug candidates for potential drug-drug interactions with OATP1B1 and OATP1B3.
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