Assessing Trans-Inhibition of OATP1B1 and OATP1B3 by Calcineurin and/or PPIase Inhibitors and Global Identification of OATP1B1/3-Associated Proteins.
Assessing Trans-Inhibition of OATP1B1 and OATP1B3 by Calcineurin and/or PPIase Inhibitors and Global Identification of OATP1B1/3-Associated Proteins.
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DOI：
10.3390/pharmaceutics16010063
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发表时间：
2023-12-31
期刊：
影响因子：
5.4
通讯作者：
Yue W
中科院分区：
文献类型：
--
作者：
Powell JT;Kayesh R;Ballesteros-Perez A;Alam K;Niyonshuti P;Soderblom EJ;Ding K;Xu C;Yue W
有机阴离子转运多肽（OATP） 1B1和OATP1B3是药物-药物相互作用（ddi）的关键决定因素。包括钙调磷酸酶抑制剂（CNI）环孢素A （CsA）在内的多种药物通过未知机制对OATP1B1和/或OATP1B3（简称OATP1B1/3）产生预孵育诱导的反式抑制作用。OATP1B1/3是磷酸化蛋白；calcalineurin能够去磷酸化并调节许多磷酸化蛋白，但在预孵育诱导的OATP1B1/3反式抑制中尚未被研究。在此，我们比较了CsA、非CNI CsA类似物他克莫司和非CNI CsA类似物SCY-635在转运蛋白过表达的人胚胎肾（HEK） 293稳定细胞系中对OATP1B1和OATP1B3的反式抑制作用。与对照相比，他克莫司（1-10µM）预孵卵（10-60 min）可迅速显著降低OATP1B1-和oatp1b3介导的转运，分别为0.18±0.03和0.20±0.02倍。CsA和SCY-635均能反式抑制OATP1B1，在60 min的预孵育时间内，抑制作用逐渐增强。在每个等效的预孵育时间，CsA对OATP1B1的反式抑制作用大于SCY-635。SCY-635预孵育60 min，对OATP1B1的IC50为2.2±1.4µM，是CsA（0.12±0.04µM）的18倍。此外，基于蛋白质组学的蛋白质相互作用筛选用于检查可能参与OATP1B1/3调控和CNIs和其他药物诱导的预孵育抑制的蛋白质和过程。共鉴定出861和357个蛋白分别与OATP1B1和OATP1B3特异性相关，包括各种蛋白激酶、泛素相关酶、他克莫司（FK506）结合蛋白FKBP5和FKBP8，以及几个已知的钙调磷酸酶调节靶点。目前的研究报告了一些新的发现，扩大了我们对OATP1B1/3功能受损的理解；其中包括：CNI他克莫斯对OATP1B1/3的预培养诱导的反式抑制作用，与非CNI类似物SCY-635相比，CsA对OATP1B1/3的预培养诱导的抑制作用更大，以及OATP1B1/3与各种与已建立和候选OATP1B1/3调节过程相关的蛋白质的关联。
Organic anion transporting polypeptide (OATP) 1B1 and OATP1B3 are key determinants of drug–drug interactions (DDIs). Various drugs including the calcineurin inhibitor (CNI) cyclosporine A (CsA) exert preincubation-induced trans-inhibitory effects upon OATP1B1 and/or OATP1B3 (abbreviated as OATP1B1/3) by unknown mechanism(s). OATP1B1/3 are phosphoproteins; calcineurin, which dephosphorylates and regulates numerous phosphoproteins, has not previously been investigated in the context of preincubation-induced trans-inhibition of OATP1B1/3. Herein, we compare the trans-inhibitory effects exerted on OATP1B1 and OATP1B3 by CsA, the non-analogous CNI tacrolimus, and the non-CNI CsA analogue SCY-635 in transporter-overexpressing human embryonic kidney (HEK) 293 stable cell lines. Preincubation (10–60 min) with tacrolimus (1–10 µM) rapidly and significantly reduces OATP1B1- and OATP1B3-mediated transport up to 0.18 ± 0.03- and 0.20 ± 0.02-fold compared to the control, respectively. Both CsA and SCY-635 can trans-inhibit OATP1B1, with the inhibitory effects progressively increasing over a 60 min preincubation time. At each equivalent preincubation time, CsA has greater trans-inhibitory effects toward OATP1B1 than SCY-635. Preincubation with SCY-635 for 60 min yielded IC50 of 2.2 ± 1.4 µM against OATP1B1, which is ~18 fold greater than that of CsA (0.12 ± 0.04 µM). Furthermore, a proteomics-based screening for protein interactors was used to examine possible proteins and processes contributing to OATP1B1/3 regulation and preincubation-induced inhibition by CNIs and other drugs. A total of 861 and 357 proteins were identified as specifically associated with OATP1B1 and OATP1B3, respectively, including various protein kinases, ubiquitin-related enzymes, the tacrolimus (FK506)-binding proteins FKBP5 and FKBP8, and several known regulatory targets of calcineurin. The current study reports several novel findings that expand our understanding of impaired OATP1B1/3 function; these include preincubation-induced trans-inhibition of OATP1B1/3 by the CNI tacrolimus, greater preincubation-induced inhibition by CsA compared to its non-CNI analogue SCY-635, and association of OATP1B1/3 with various proteins relevant to established and candidate OATP1B1/3 regulatory processes.
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影响因子：
2.9
作者：
Daum, Sebastian;Schumann, Michael;Mathea, Sebastian;Aumueller, Tobias;Balsley, Molly A.;Constant, Stephanie L.;de Lacroix, Boris Feaux;Kruska, Fabian;Braun, Manfred;Schiene-Fischer, Cordelia
通讯作者：
Schiene-Fischer, Cordelia
影响因子：
3.7
作者：
Crowe A;Zheng W;Miller J;Pahwa S;Alam K;Fung KM;Rubin E;Yin F;Ding K;Yue W
通讯作者：
Yue W
影响因子：
7.3
作者：
Fu, Jiping;Tjandra, Meiliana;Sweeney, Zachary K.
通讯作者：
Sweeney, Zachary K.
影响因子：
32.4
作者：
Colgan, J;Asmal, M;Luban, J
通讯作者：
Luban, J
影响因子：
3.8
作者：
Farasyn, Taleah;Crowe, Alexandra;Yue, Wei
通讯作者：
Yue, Wei