Catalytic inhibition of topoisomerase II by a novel rationally designed ATP-competitive purine analogue.
Catalytic inhibition of topoisomerase II by a novel rationally designed ATP-competitive purine analogue.
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DOI：
10.1186/1472-6769-9-1
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发表时间：
2009-01-07
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拓扑异构酶II毒药作为抗癌治疗已经临床使用了几十年，它通过稳定酶诱导的DNA断裂来发挥作用。相比之下，催化抑制剂在DNA断裂之前阻止该酶。虽然已经描述了几种拓扑异构酶II的催化抑制剂，但基于肿瘤细胞的分子特征来开发其抗增殖活性的临床前概念直到最近才开始出现。拓扑异构酶II是一种ATPase，它利用ATP水解产生的能量来协调DNA双链沿该酶的运动。因此，以核苷酸结合口袋为靶点的低分子量抑制剂干扰ATPase功能，应该会对致力于进行有丝分裂的细胞产生深远的影响。在这里，我们描述了一种新的嘌呤二胺类似物的发现和表征，它是一种有效的拓扑异构酶II的ATP竞争性催化抑制剂。喹啉氨基嘌呤化合物1(QAP 1)在亚微摩尔浓度下抑制拓扑异构酶II的ATPase活性和去甲基化反应，在无细胞检测中靶向于拓扑异构酶IIα和β，并通过基于细胞的定量分析和染色体分离试验，在细胞中显示出催化酶抑制。与最近的假设一致，我们发现BRCA1突变的乳腺癌细胞对QAP 1的敏感性增加。QAP 1的结果表明，用ATP竞争性抑制剂有效和选择性地催化抑制人拓扑异构酶II功能是可行的。我们的数据表明，有必要对ATP竞争性催化抑制剂进行进一步的药物发现，并且此类药物可能被开发为具有适当分子改变组合的肿瘤的抗癌治疗。
Topoisomerase II poisons are in clinical use as anti-cancer therapy for decades and work by stabilizing the enzyme-induced DNA breaks. In contrast, catalytic inhibitors block the enzyme before DNA scission. Although several catalytic inhibitors of topoisomerase II have been described, preclinical concepts for exploiting their anti-proliferative activity based on molecular characteristics of the tumor cell have only recently started to emerge. Topoisomerase II is an ATPase and uses the energy derived from ATP hydrolysis to orchestrate the movement of the DNA double strands along the enzyme. Thus, interfering with ATPase function with low molecular weight inhibitors that target the nucleotide binding pocket should profoundly affect cells that are committed to undergo mitosis. Here we describe the discovery and characterization of a novel purine diamine analogue as a potent ATP-competitive catalytic inhibitor of topoisomerase II. Quinoline aminopurine compound 1 (QAP 1) inhibited topoisomerase II ATPase activity and decatenation reaction at sub-micromolar concentrations, targeted both topoisomerase II alpha and beta in cell free assays and, using a quantitative cell-based assay and a chromosome segregation assay, displayed catalytic enzyme inhibition in cells. In agreement with recent hypothesis, we show that BRCA1 mutant breast cancer cells have increased sensitivity to QAP 1. The results obtained with QAP 1 demonstrate that potent and selective catalytic inhibition of human topoisomerase II function with an ATP-competitive inhibitor is feasible. Our data suggest that further drug discovery efforts on ATP-competitive catalytic inhibitors are warranted and that such drugs could potentially be developed as anti-cancer therapy for tumors that bear the appropriate combination of molecular alterations.