Discovery and characterization of novel small molecule inhibitors of human Cdc25B dual specificity phosphatase
Discovery and characterization of novel small molecule inhibitors of human Cdc25B dual specificity phosphatase
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DOI：
10.1124/mol.104.001784
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发表时间：
2004-10-01
影响因子：
3.6
通讯作者：
Lazo, JS
中科院分区：
文献类型：
--
作者：
Brisson, M;Nguyen, T;Lazo, JS
Cdc 25 A和Cdc 25 B双特异性磷酸酶是细胞周期转换和增殖的关键调节因子。它们具有致癌特性并且在许多人类肿瘤中过表达。因为选择性Cdc 25磷酸酶抑制剂将是有价值的生物工具和可能的治疗剂，我们已经测定了小分子库在体外抑制Cdc 25。我们现在报告的两个新的结构不同类的Cdc 25抑制剂与细胞活性的鉴定。环戊喹啉3a，4，5，9 b-四氢-3H-环戊[c]喹啉-4，8-二羧酸（5661118）和萘并呋喃二酮3-苯甲酰基-萘并[1，2-B]呋喃-4，5-二酮（5169131）对重组Cdc 25的体外IC 50值为2.5至11 μ M，对其他磷酸酶的抑制效力较低。与5661118不同，5169131对Cdc 25 B产生可逆抑制，并显示竞争性抑制剂动力学。用5661118没有观察到生长抑制活性，而10至30 μ M 5169131引起G（1）/S和G（2）/M停滞。我们还发现5169131抑制人PC-3前列腺癌和MDA-MB-435乳腺癌细胞增殖。浓度依赖性Tyr 15过度磷酸化被视为对细胞周期蛋白依赖性激酶与1小时5169131治疗，与Cdc 25抑制一致。耐DNA拓扑异构酶II抑制剂的细胞对5169131的敏感性与亲本细胞一样，表明这种醌化合物在体内不抑制拓扑异构酶II。分子建模用于预测抑制剂和Cdc 25 B之间的潜在相互作用位点，并提供有关实验观察的分子起源的见解。基于其动力学特征和细胞活性，我们认为5169131可能是一个很好的工具，为进一步研究Cdc 25的细胞作用。
Cdc25A and Cdc25B dual-specificity phosphatases are key regulators of cell cycle transition and proliferation. They have oncogenic properties and are overexpressed in many human tumors. Because selective Cdc25 phosphatase inhibitors would be valuable biological tools and possible therapeutic agents, we have assayed a small molecule library for in vitro inhibition of Cdc25. We now report the identification of two new structurally distinct classes of Cdc25 inhibitors with cellular activity. The cyclopentaquinoline 3a, 4,5,9b-tetrahydro-3H-cyclopenta[c] quinoline-4,8-dicarboxylic acid (5661118) and the naphthofurandione 3-benzoyl-naphtho[1,2-b] furan-4,5-dione (5169131) had in vitro IC50 values of 2.5 to 11 muM against recombinant Cdc25 and were less potent inhibitors of other phosphatases. Unlike 5661118, 5169131 caused reversible inhibition of Cdc25B and displayed competitive inhibitor kinetics. No growth inhibitory activity was seen with 5661118, whereas 10 to 30 muM 5169131 caused G(1)/S and G(2)/M arrest. We also found that 5169131 inhibited human PC-3 prostate and MDA-MB-435 breast cancer cell proliferation. Concentration-dependent Tyr15 hyperphosphorylation was seen on cyclin-dependent kinase with a 1-h 5169131 treatment, consistent with Cdc25 inhibition. Cells resistant to DNA toposiomerase II inhibitors were as sensitive to 5169131 as parental cells, indicating that this quinone compound does not inhibit topoisomerase II in vivo. Molecular modeling was used to predict a potential interaction site between the inhibitor and Cdc25B and to provide insights as to the molecular origins of the experimental observations. Based on its kinetic profile and cellular activity, we suggest that 5169131 could be an excellent tool for further studies on the cellular roles of Cdc25.