Characterization of Aripiprazole Uptake Transporter in the Blood-Brain Barrier Model hCMEC/D3 Cells by Targeted siRNA Screening
Characterization of Aripiprazole Uptake Transporter in the Blood-Brain Barrier Model hCMEC/D3 Cells by Targeted siRNA Screening
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通过靶向 siRNA 筛选表征血脑屏障模型 hCMEC/D3 细胞中的阿立哌唑摄取转运蛋白
DOI：
10.1007/s11095-022-03223-z
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发表时间：
2022
影响因子：
3.7
通讯作者：
Ikumi Tamai
中科院分区：
文献类型：
--
作者：
Moeno Kadoguchi;Hiroshi Arakawa;Ryokichi Honda;Kazuki Hotta;Yoshiyuki Shirasaka;Yoshiharu Deguchi;Ikumi Tamai
目的血脑屏障（BBB）摄取转运蛋白的鉴定是中枢神经系统（CNS）药物研发中的一个重大挑战。然而，考虑已知药物摄取特征的常规方法未能识别负责的转运蛋白分子。本研究的目的是确定阿立哌唑摄取转运BBB模型hCMEC/D3细胞使用敲低筛选研究针对各种转运蛋白，包括uncharacterized ons.MethodsWe评估了214种类型的siRNA靶向转运蛋白的影响，阿立哌唑，非典型抗精神病药物，在hCMEC/D3细胞的摄取。阿立哌唑的摄取测定usingXenopusoocytes表达的候选基因提取的siRNA screening assay.ResultsThe估计未结合的脑血浆浓度比（KP，UU，脑）的阿立哌唑估计为0.67野生型小鼠和1.94 inabcb 1a/1b/abcg 2敲除小鼠，这表明在BBB渗透的摄取和外排转运蛋白的参与。根据siRNA敲除筛选研究，有机阳离子/肉毒碱转运蛋白2（OCTN 2）和长链脂肪酸转运蛋白1（FATP 1）被确定为候选基因。OCTN 2抑制剂可降低hCMEC/D3细胞对阿立哌唑的摄取，而FATP 1抑制剂则无此作用。在表达OCTN 2的异种卵母细胞中观察到阿立哌唑的摄入部分增加。最后，评估转运蛋白介导的血脑屏障渗透的药物，报告和estimatedKp，uu，brainvalues were summarized.ConclusionsA knockdown筛选研究结合与Kp，uu，brainvalues表明，阿立哌唑是一个潜在的底物OCTN 2。本研究中所描述的技术可用于鉴定CNS药物的新型BBB转运体。
AimIdentification of blood-brain barrier (BBB) uptake transporters is a major challenge in the research and development of central nervous system (CNS) drugs. However, conventional methods that consider known drug uptake characteristics have failed at identifying the responsible transporter molecule. The present study aimed at identifying aripiprazole uptake transporters in BBB model hCMEC/D3 cells using a knockdown screening study targeting various transporters, including uncharacterized ones.MethodsWe evaluated the effect of 214 types of siRNA targeting transporters on the uptake of aripiprazole, an atypical antipsychotic drug, in hCMEC/D3 cells. Aripiprazole uptake was determined usingXenopusoocytes expressing the candidate genes extracted from the siRNA screening assay.ResultsThe estimated unbound brain to plasma concentration ratio (Kp,uu,brain) of aripiprazole was estimated as 0.67 in wild-type mice and 1.94 inabcb1a/1b/abcg2knockout mice, suggesting the involvement of both uptake and efflux transporters in BBB permeation. According to siRNA knockdown screening studies, organic cation/carnitine transporter 2 (OCTN2) and long-chain fatty acid transporter 1 (FATP1) were identified as candidate genes. The uptake of aripiprazole by hCMEC/D3 cells was decreased by OCTN2 inhibitors, but not by FATP1 inhibitors. A partially increased uptake of aripiprazole was observed in OCTN2-expressingXenopusoocytes. Finally, to evaluate transporter-mediated BBB permeation of drugs, the reported and estimatedKp,uu,brainvalues were summarized.ConclusionsA knockdown screening study in combination withKp,uu,brainvalues showed that aripiprazole was a potential substrate of OCTN2. The technique described in this study can be applied to identifying novel BBB transporters for CNS drugs.
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DOI：
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发表时间：
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期刊：
影响因子：
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作者：
通讯作者：
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影响因子：
3.6
作者：
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通讯作者：
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DOI：
10.1124/jpet.116.232447
发表时间：
2016-08-01
影响因子：
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作者：
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影响因子：
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作者：
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通讯作者：
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