Drug-Resistant Aurora A Mutants for Cellular Target Validation of the Small Molecule Kinase Inhibitors MLN8054 and MLN8237
Drug-Resistant Aurora A Mutants for Cellular Target Validation of the Small Molecule Kinase Inhibitors MLN8054 and MLN8237
复制标题
DOI：
10.1021/cb100053q
复制
发表时间：
2010-06-01
影响因子：
4
通讯作者：
Eyers, Patrick A.
中科院分区：
文献类型：
--
作者：
Sloane, Dominic A.;Trikic, Michael Z.;Eyers, Patrick A.
Aurora激酶调节有丝分裂进程的多个方面，它们在不同肿瘤类型中的过表达使它们成为有吸引力的肿瘤学靶点。在过去的十年中，密集的研究工作已经导致发现了化学上不同的小分子极光激酶抑制剂家族，其中许多已经在模型系统中显示出治疗潜力。这些药物也是帮助剖析由Aurora激酶协调的信号通路的重要工具，并且泛Aurora抑制剂如VX-680的抗增殖靶点已经使用化学遗传技术进行了验证。在许多情况下，Aurora抑制剂对不相关激酶的非特异性已经得到了很好的证实，这可能会扩大这些化合物可能适用的癌症范围。然而，明确地证明临床激酶抑制剂的分子靶点是一个重要的挑战，这对于解释化合物特异性、耐药性和疗效的分子基础是绝对关键的。在本文中，我们研究了Aurora A对基于苯并氮杂卓的Aurora抑制剂MLN 8054和类似物MLN 8237（一种处于II期临床试验的第二代化合物）敏感性的氨基酸需求。晶体学分析促进了一组耐药Aurora A突变体的设计和生化研究，然后选择其中的一个子集作为候选耐药靶点进行进一步评价。使用诱导型人类细胞系，我们发现表达接近生理水平的功能性但部分耐药的Aurora A T217 D突变体的细胞在MLN 8054或MLN 8237存在下存活，从而验证Aurora A是这些化合物的关键抗增殖靶点。
The Aurora kinases regulate multiple aspects of mitotic progression, and their overexpression in diverse tumor types makes them appealing oncology targets. An intensive research effort over the past decade has led to the discovery of chemically distinct families of small molecule Aurora kinase inhibitors, many of which have demonstrated therapeutic potential in model systems. These agents are also important tools to help dissect signaling pathways that are orchestrated by Aurora kinases, and the antiproliferative target of pan-Aurora inhibitors such as VX-680 has been validated using chemical genetic techniques. In many cases the nonspecific nature of Aurora inhibitors toward unrelated kinases is well established, potentially broadening the spectrum of cancers to which these compounds might be applied. However, unambiguously demonstrating the molecular target(s) for clinical kinase inhibitors is an important challenge, one that is absolutely critical for deciphering the molecular basis of compound specificity, resistance, and efficacy. In this paper, we have investigated amino acid requirements for Aurora A sensitivity to the benzazepine-based Aurora inhibitor MLN8054 and the close analogue MLN8237, a second-generation compound that is in phase II clinical trials. A crystallographic analysis facilitated the design and biochemical investigation of a panel of resistant Aurora A mutants, a subset of which were then selected as candidate drug-resistance targets for further evaluation. Using inducible human cell lines, we show that cells expressing near-physiological levels of a functional but partially drug-resistant Aurora A T217D mutant survive in the presence of MLN8054 or MLN8237, authenticating Aurora A as a critical antiproliferative target of these compounds.