Fluorescent substrates for flow cytometric evaluation of efflux inhibition in ABCB1, ABCC1, and ABCG2 transporters.
Fluorescent substrates for flow cytometric evaluation of efflux inhibition in ABCB1, ABCC1, and ABCG2 transporters.
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DOI：
10.1016/j.ab.2013.02.018
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发表时间：
2013-06-01
影响因子：
2.9
通讯作者：
Sklar LA
中科院分区：
文献类型：
--
作者：
Strouse JJ;Ivnitski-Steele I;Waller A;Young SM;Perez D;Evangelisti AM;Ursu O;Bologa CG;Carter MB;Salas VM;Tegos G;Larson RS;Oprea TI;Edwards BS;Sklar LA
ATP结合盒（ABC）跨膜外排泵如P-糖蛋白（ABCB 1）、多药耐药蛋白1（ABCC 1）和乳腺癌耐药蛋白（ABCG 2）在抗癌药物耐药中起重要作用。大量结构和功能不同的化合物作为这些泵的底物或调节剂。体外评价候选药物对多药耐药蛋白的亲和力是预测体内药代动力学和药物-药物相互作用的关键。本研究的目的是确定和表征这些转运蛋白的新底物。作为与Life Technologies合作项目的一部分，在ABC转运蛋白的流式细胞仪筛选中研究了102种荧光探针。初步筛选比较了亲本细胞系与其相应的高表达抗性对应物中的底物外排活性。荧光化合物库包括一系列激发/发射曲线，并且需要双激光激发以及多个荧光检测通道。共鉴定了31种在一个或多个泵中具有主动外排和实际荧光响应范围的底物，并测试了其与8种已知抑制剂的相互作用。这种筛选方法提供了一种有效的工具，用于识别和表征ABCB 1，ABCC 1和ABCG 2的新荧光底物。
ATP binding cassette (ABC) transmembrane efflux pumps such as P-glycoprotein (ABCB1), multidrug resistance protein 1 (ABCC1), and breast cancer resistance protein (ABCG2) play an important role in anti-cancer drug resistance. A large number of structurally and functionally diverse compounds act as substrates or modulators of these pumps. In vitro assessment of the affinity of drug candidates for multidrug resistance proteins is central to predict in vivo pharmacokinetics and drug–drug interactions. The objective of this study was to identify and characterize new substrates for these transporters. As part of a collaborative project with Life Technologies, 102 fluorescent probes were investigated in a flow cytometric screen of ABC transporters. The primary screen compared substrate efflux activity in parental cell lines with their corresponding highly expressing resistant counterparts. The fluorescent compound library included a range of excitation/emission profiles and required dual laser excitation as well as multiple fluorescence detection channels. A total of 31 substrates with active efflux in one or more pumps and practical fluorescence response ranges were identified and tested for interaction with eight known inhibitors. This screening approach provides an efficient tool for identification and characterization of new fluorescent substrates for ABCB1, ABCC1, and ABCG2.
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影响因子：
3.7
作者：
Lebedeva IV;Pande P;Patton WF
通讯作者：
Patton WF
影响因子：
5.7
作者：
Henrich, Curtis J.;Robey, Robert W.;McMahon, James B.
通讯作者：
McMahon, James B.
影响因子：
3.4
作者：
Özvegy-Lacka, C;Köblös, G;Váradi, A
通讯作者：
Váradi, A
DOI：
10.1126/science.1168750
发表时间：
2009-03-27
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Aller SG;Yu J;Ward A;Weng Y;Chittaboina S;Zhuo R;Harrell PM;Trinh YT;Zhang Q;Urbatsch IL;Chang G
通讯作者：
Chang G
影响因子：
3.6
作者：
Martin, C;Berridge, G;Callaghan, R
通讯作者：
Callaghan, R