A Systematic In Vitro Investigation of the Inhibitor Preincubation Effect on Multiple Classes of Clinically Relevant Transporters
A Systematic In Vitro Investigation of the Inhibitor Preincubation Effect on Multiple Classes of Clinically Relevant Transporters
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抑制剂预孵育对多类临床相关转运蛋白影响的系统体外研究
DOI：
10.1124/dmd.118.085993
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发表时间：
2019
影响因子：
3.9
通讯作者：
F. Huth
中科院分区：
文献类型：
--
作者：
Péter Tátrai;Patrick Schweigler;B. Poller;Norbert Domange;R. D. de Wilde;Imad Hanna;Zsuzsanna Gáborik;F. Huth
在基于细胞的测定中，药物转运蛋白与其抑制剂的预孵育可能导致抑制效力的明显增强。目前，关于预孵育（PTIP）增强除OATP 1B 1和OATP 1B 3以外的临床相关溶质载体转运蛋白的转运蛋白抑制作用的数据有限。因此，使用OATP 1B 1、OATP 1B 3、OAT 1、OAT 3、OCT 1、OCT 2、MATE 1和MATE 2-K细胞系系统检查PTIP。在预孵育或未预孵育3小时的情况下测定30种抑制剂的IC 50值，并通过评估转运抑制效力和细胞浓度的时间过程进一步表征PTIP ≥2.5×的化合物。对于每种化合物，计算观察到的最高PTIP与物理化学性质之间的相关性。PTIP在有机阳离子转运蛋白（OCT）和有机阴离子转运多肽（OATP）中普遍存在，但在有机阴离子转运蛋白（OAT）或多药和毒素外排转运蛋白（MATE）中不存在，并且在控制毒性和非特异性结合后，大多数PTIP病例仍持续存在。偶尔，需要预孵育超过2小时以达到完全抑制效力。对于所检查的四种药物，根据现行监管标准，预孵育有可能将体外药物相互作用风险预测从“无风险”变为“风险”。分子量和LogD 7.4以及被动细胞蓄积和细胞摄取速率的比值与PTIP相关;因此，低细胞渗透和未结合细胞内抑制剂浓度的缓慢积累可能导致PTIP。综上所述，我们的数据表明，PTIP部分由肇事药物的理化性质决定，预孵育可能会影响OCT和OATP的体外预测药物相互作用风险。重要性声明在新药开发过程中，进行体外研究以评估患者可能已经服用的现有药物与新型化合物之间潜在不良相互作用的风险。进行这些体外试验的确切方式可能会影响风险评估的结果。在这里，我们认为，相互作用的风险可能被低估，除非特定的试验方案进行修改，包括一个额外的孵育步骤，允许测试药物在细胞内积累，并证明，增加这一步骤是特别重要的大和疏水性的药物分子。
Preincubation of a drug transporter with its inhibitor in a cell-based assay may result in the apparent enhancement of the inhibitory potency. Currently, limited data are available on potentiation of transporter inhibition by preincubation (PTIP) for clinically relevant solute-carrier transporters other than OATP1B1 and OATP1B3. Therefore, PTIP was examined systematically using OATP1B1, OATP1B3, OAT1, OAT3, OCT1, OCT2, MATE1, and MATE2-K cell lines. IC50 values of 30 inhibitors were determined with or without 3 hours of preincubation, and compounds with a PTIP ≥2.5× were further characterized by assessing the time course of transport inhibition potency and cellular concentration. For each compound, correlations were calculated between highest observed PTIP and physicochemical properties. PTIP was prevalent among organic cation transporters (OCTs) and organic anion–transporting polypeptides (OATPs) but not among organic anion transporters (OATs) or multidrug and toxin extrusion transporters (MATEs), and most instances of PTIP persisted after controlling for toxicity and nonspecific binding. Occasionally, preincubation in excess of 2 hours was required to attain full inhibitory potency. For four drugs examined, preincubation had the potential to change the in vitro drug-drug interaction risk prediction from “no risk” to “risk” on the basis of current regulatory criteria. Molecular weight and LogD7.4, as well as the ratio of passive cellular accumulation and cellular uptake rate correlated with PTIP; thus, low cellular permeation and a slow build-up of unbound intracellular inhibitor concentration may contribute to PTIP. Taken together, our data suggest that PTIP is partly determined by the physicochemical properties of the perpetrator drug, and preincubation may affect the in vitro predicted drug-drug interaction risk for OCTs as well as OATPs. Significance Statement During the development of a novel pharmaceutical drug, in vitro studies are conducted to assess the risk of potential adverse interactions between existing medications a patient may already be taking and the novel compound. The exact way these in vitro assays are performed may influence the outcome of risk assessment. Here we suggest that the interaction risk may be underestimated unless specific assay protocols are modified to include an additional incubation step that allows the test drug to accumulate inside the cells, and demonstrate that adding this step is particularly important for large and hydrophobic drug molecules.
影响因子：
3.9
作者：
Hirano, Masaru;Maeda, Kazuya;Sugiyama, Yuichi
通讯作者：
Sugiyama, Yuichi
环孢素 A 和他克莫司对 OATP1B1 和 OATP1B3 介导的摄取的比较抑制作用
DOI：
--
发表时间：
2009
期刊：
影响因子：
--
作者：
設楽悦久;竹内久美子;和田怜美;永松佳子;杉山雄一;堀江利治
通讯作者：
堀江利治