Abstract

1. Interactions of the Janus kinase (JAK) inhibitor ruxolitinib with solute carriers (SLCs) remain incompletely characterised. The present study was therefore designed to investigate this issue.

2. The interactions of ruxolitinib with SLCs were analysed using transporter-overexpressing human embryonic kidney HEK293 cells. Substrate accumulation was detected by spectrofluorimetry, liquid chromatography coupled to tandem mass spectrometry or scintillation counting.

3. Ruxolitinib was found to potently inhibit the activities of organic anion transporter 3 (OAT3), organic cation transporter 2 (OCT2), multidrug and toxin extrusion 1 (MATE1) and MATE2-K (half maximal inhibitory concentration (IC₅₀) < 10 µM). It blocked OAT1, OAT4, OATP1B1, OATP1B3, OATP2B1 and OCT3, but in a weaker manner (IC₅₀ > 10 µM), whereas OCT1 was not impacted. No time-dependent inhibition was highlighted. When applying the US Food and Drug Administration (FDA) criteria for transporters-related drug-drug interaction risk, OCT2 and MATE2-K, unlike MATE1 and OAT3, were predicted to be in vivo inhibited by ruxolitinib. Cellular uptake studies additionally indicated that ruxolitinib is a substrate for MATE1 and MATE2-K, but not for OAT3 and OCT2.

4. Ruxolitinib in vitro blocked activities of most of SLC transporters. Only OCT2 and MATE-2K may be however clinically inhibited by the JAK inhibitor, with the caution for OCT2 that in vitro inhibition data were generated with an FDA-non recommended fluorescent substrate. Ruxolitinib MATEs-mediated transport may additionally deserve attention for its possible pharmacological consequences in MATE-positive cells.

摘要

1. Janus 激酶 (JAK) 抑制剂鲁索替尼与溶质载体 (SLC) 的相互作用仍未完全表征。因此，本研究旨在调查这个问题。

2. 使用过表达转运蛋白的人胚胎肾 HEK293 细胞分析鲁索替尼与 SLC 的相互作用。通过荧光分光光度法、液相色谱法与串联质谱法或闪烁计数法检测底物积累。

3. 发现 Ruxolitinib 可有效抑制有机阴离子转运蛋白 3 (OAT3)、有机阳离子转运蛋白 2 (OCT2)、多药和毒素挤出 1 (MATE1) 和 MATE2-K（半数最大抑制浓度 (IC ₅₀ ) < 10 µM）的活性. 它阻断了 OAT1、OAT4、OATP1B1、OATP1B3、OATP2B1 和 OCT3，但作用较弱（IC ₅₀ > 10 µM），而 OCT1 没有受到影响。没有突出显示时间依赖性抑制。当应用美国食品和药物管理局 (FDA) 的转运蛋白相关药物相互作用风险标准时，与 MATE1 和 OAT3 不同，OCT2 和 MATE2-K 被预测会在体内被鲁索替尼抑制。细胞摄取研究还表明，鲁索替尼是 MATE1 和 MATE2-K 的底物，但不是 OAT3 和 OCT2 的底物。

4. Ruxolitinib在体外阻断了大多数 SLC 转运蛋白的活性。然而，只有 OCT2 和 MATE-2K 可能会被 JAK 抑制剂在临床上抑制，注意 OCT2的体外抑制数据是用 FDA 非推荐的荧光底物产生的。Ruxolitinib MATEs 介导的转运可能因其在 MATE 阳性细胞中可能产生的药理学后果而值得关注。