Differential Interaction of Dantrolene, Glafenine, Nalidixic Acid, and Prazosin with Human Organic Anion Transporters 1 and 3
Differential Interaction of Dantrolene, Glafenine, Nalidixic Acid, and Prazosin with Human Organic Anion Transporters 1 and 3
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DOI：
10.1124/jpet.117.241406
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发表时间：
2017-09
期刊：
影响因子：
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通讯作者：
B. Burckhardt;M. Henjakovic;Y. Hagos;G. Burckhardt
中科院分区：
文献类型：
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作者：
B. Burckhardt;M. Henjakovic;Y. Hagos;G. Burckhardt
在肾近端小管细胞中，基底外侧膜中的有机阴离子转运蛋白1和3（OAT 1和OAT 3）和顶膜中的多药耐药相关蛋白4（MRP 4）共享底物并在肾脏药物分泌中合作。我们假设，最近发现的MRP 4抑制剂丹曲林，格拉非宁，萘地酸，哌唑嗪也与人OAT 1和/或OAT 3在人胚肾293细胞中稳定转染相互作用。这四种药物被测试为OAT 1摄取p-[3 H]氨基马尿酸盐（PAH）和[14 C]戊二酸盐以及OAT 3摄取[3 H]雌酮-3-硫酸盐（ES）的可能抑制剂。此外，我们还探讨了这些药物是否会降低放射性标记的PAH、戊二酸盐或ES的平衡分布，这是一种旨在间接表明通过OAT 1和OAT 3进行药物/底物交换的方法。使用OAT 3时，观察到[3 H]ES摄取的剂量依赖性抑制和[3 H]ES平衡的下移，表明所有4种药物均与OAT 3结合，并可能发生易位。相反，与OAT 1的相互作用更为复杂。以[14 C]戊二酸为底物，所有四种药物均抑制摄取，但只有格拉非宁和萘二甲酸改变戊二酸平衡。使用[3 H]PAH作为OAT 1的底物，萘地平酸抑制但丹曲林、格拉非宁和哌唑嗪刺激摄取。萘啶酸降低了[3 H]PAH的平衡含量，这表明它可能被OAT 1交换。总之，OAT 1和OAT 3与MRP 4抑制剂丹曲林、格拉非宁、萘地酸和哌唑嗪相互作用，表明特异性重叠。在OAT 1处，必须假设有一个以上的结合位点，以解释底物和药物依赖性刺激和抑制转运活性。
In renal proximal tubule cells, the organic anion transporters 1 and 3 (OAT1 and OAT3) in the basolateral membrane and the multidrug resistance-associated protein 4 (MRP4) in the apical membrane share substrates and co-operate in renal drug secretion. We hypothesized that recently identified MRP4 inhibitors dantrolene, glafenine, nalidixic acid, and prazosin also interact with human OAT1 and/or OAT3 stably transfected in human embryonic kidney 293 cells. These four drugs were tested as possible inhibitors of p-[3H]aminohippurate (PAH) and [14C]glutarate uptake by OAT1, and of [3H]estrone-3-sulfate (ES) uptake by OAT3. In addition, we explored whether these drugs decrease the equilibrium distribution of radiolabeled PAH, glutarate, or ES, an approach intended to indirectly suggest drug/substrate exchange through OAT1 and OAT3. With OAT3, a dose-dependent inhibition of [3H]ES uptake and a downward shift in [3H]ES equilibrium were observed, indicating that all four drugs bind to OAT3 and may possibly be translocated. In contrast, the interaction with OAT1 was more complex. With [14C]glutarate as substrate, all four drugs inhibited uptake but only glafenine and nalidixic acid shifted glutarate equilibrium. Using [3H]PAH as a substrate of OAT1, nalidixic acid inhibited but dantrolene, glafenine, and prazosin stimulated uptake. Nalidixic acid decreased equilibrium content of [3H]PAH, suggesting that it may possibly be exchanged by OAT1. Taken together, OAT1 and OAT3 interact with the MRP4 inhibitors dantrolene, glafenine, nalidixic acid, and prazosin, indicating overlapping specificities. At OAT1, more than one binding site must be assumed to explain substrate and drug-dependent stimulation and inhibition of transport activity.