New inhibitors of ABCG2 identified by high-throughput screening
New inhibitors of ABCG2 identified by high-throughput screening
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DOI：
10.1158/1535-7163.mct-07-0352
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发表时间：
2007-12-01
影响因子：
5.7
通讯作者：
McMahon, James B.
中科院分区：
文献类型：
--
作者：
Henrich, Curtis J.;Robey, Robert W.;McMahon, James B.
为了寻找新的三磷酸腺苷结合盒转运体(ABCG2)的抑制剂，采用高通量方法检测ABCG2底物脱镁叶绿酸a在ABCG2过表达的NCI-H460 MX20细胞中的积累，以筛选化合物文库。从国家癌症研究所/发展治疗计划收集的7,325个天然产物和合成化合物库中，发现18个化合物在10MU/L时对ABCG2具有抑制作用，在从18个原始化合物中去除类黄酮类化合物和有限可用化合物后，剩下的11个化合物中有10个化合物逆转了NCI-H460/MX20细胞对米托蒽醌的耐药性，并阻止了ABCG2介导的BODIPY-PZ在ABCG2转基因的HEK293细胞中的转运，证实了与ABCG2的相互作用。根据活性图谱和材料的可获得性，考察了五种抑制剂与ABCG2的[1251]碘芳基氮杂咪唑标记、增加抗ABCG2抗体5D3的结合以及阻止P-糖蛋白或多药耐药蛋白1介导的转运的能力。在浓度为20Mol/L时，所有化合物均可使碘代芳基氮唑啉的标记率降低50%~80%。所有五个化合物还增加了ABCG2的5133个标记，表明这些化合物是ABCG2的抑制剂，但不是底物。所有化合物都不影响P-糖蛋白介导的罗丹明123转运，而三个化合物影响多药耐药蛋白-1介导的钙调素转运25[MU/L]，这表明这些化合物对ABCG2是相对特异的。这5种新的ABCG2活性抑制物可能为进一步研究ABCG2的功能及其与多药耐药的关系提供基础。
In order to identify novel inhibitors of the ATP-binding cassette transporter, ABCG2, a high-throughput assay measuring the accumulation of the ABCG2 substrate pheophorbide a in ABCG2-overexpressing NCI-H460 MX20 cells was used to screen libraries of compounds. Out of a library of 7,325 natural products and synthetic compounds from the National Cancer Institute/Developmental Therapeutics Program collection, 18 were found to inhibit ABCG2 at 10 mu mol/L. After eliminating flavonoids and compounds of limited availability from the 18 original compounds, 10 of the 11 remaining compounds reversed mitoxantrone resistance in NCI-H460/MX20 cells and prevented ABCG2-mediated BODIPY-prazosin transport in ABCG2-transfected HEK293 cells, confirming an interaction with ABCG2. Based on the activity profiles and the availability of materials, five inhibitors were examined for their ability to compete with [1251] iodoarylazidoprazosin labeling of ABCG2, increase binding of the anti-ABCG2 antibody 5D3, and prevent P-glycoprotein or multidrug resistance protein 1-mediated transport. At a concentration of 20 mu mol/L, all of the compounds reduced iodoarylazidoprazosin labeling by 50% to 80% compared with controls. All five compounds also increased 5133 labeling of ABCG2, indicating that these compounds are inhibitors but not substrates of ABCG2. None of the compounds affected P-glycoprotein-mediated rhodamine 123 transport, whereas three affected multidrug resistance protein-1-mediated calcein transport at 25 [mu mol/L, suggesting that the compounds are relatively specific for ABCG2. These five novel inhibitors of ABCG2 activity may provide a basis for further investigation of ABCG2 function and its relevance in multidrug resistance.