Targeting Drug Resistance in EGFR with Covalent Inhibitors: A Structure-Based Design Approach
Targeting Drug Resistance in EGFR with Covalent Inhibitors: A Structure-Based Design Approach
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DOI：
10.1021/acs.jmedchem.5b01082
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发表时间：
2015-09-10
影响因子：
7.3
通讯作者：
Rauh, Daniel
中科院分区：
文献类型：
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作者：
Engel, Julian;Richters, Andre;Rauh, Daniel
受体酪氨酸激酶代表癌症治疗中的主要靶标之一，因为这些细胞外信号的基本转导物（如表皮生长因子受体（EGFR））的失调有助于癌症（如非小细胞肺癌（NSCLC））的发作。大力致力于开发不可逆抑制剂，并产生了化合物CO-1686，其通过烷基化Cys 797来利用在EGFR处的停留时间增加，从而防止毒性作用。在这里，我们提出了一种基于结构的方法，合理化的随后的计算分析的构象配体合奏的解决方案，设计新的和不可逆的EGFR抑制剂的基础上筛选命中，确定在表型筛选的80个NSCLC细胞系对约1500种化合物。使用蛋白质X射线晶体学，我们破译了工程cSrc（T338 M/S345 C），一个验证的模型系统EGFR-T790 M的结合模式，这构成了进一步的合理设计方法的基础。化学合成导致进一步的化合物收集，这些化合物显示出生物化学效价增加，并且部分地对突变的（L 858 R和L 858 R/T790 M）与非突变的EGFR具有选择性。进行了进一步的基于细胞的和动力学研究以证实我们的初步发现。利用蛋白水解消化和nano-LC-MS/MS分析，我们证实了Cys 797的烷基化。
Receptor tyrosine kinases represent one of the prime targets in cancer therapy, as the dysregulation of these elementary transducers of extracellular signals, like the epidermal growth factor receptor (EGFR), contributes to the onset of cancer, such as non-small cell lung cancer (NSCLC). Strong efforts were directed to the development of irreversible inhibitors and led to compound CO-1686, which takes advantage of increased residence time at EGFR by alkylating Cys797 and thereby preventing toxic effects. Here, we present a structure-based approach, rationalized by subsequent computational analysis of conformational ligand ensembles in solution, to design novel and irreversible EGFR inhibitors based on a screening hit that was identified in a phenotype screen of 80 NSCLC cell lines against approximately 1500 compounds. Using protein X-ray crystallography, we deciphered the binding mode in engineered cSrc (T338M/S345C), a validated model system for EGFR-T790M, which constituted the basis for further rational design approaches. Chemical synthesis led to further compound collections that revealed increased biochemical potency and, in part, selectivity toward mutated (L858R and L858R/T790M) vs nonmutated EGFR. Further cell-based and kinetic studies were performed to substantiate our initial findings. Utilizing proteolytic digestion and nano-LC-MS/MS analysis, we confirmed the alkylation of Cys797.