Three types of 2-arylamino-4-(piperidin-4-yloxy)pyrimidines (I–III) were designed and synthesized as covalent EGFR(epidermal growth factor receptor)^T790M/L858R inhibitors by replacement of the common reported 4-(3-amino)phenoxyl moiety with 4-(4-hydroxy)piperidine-4-oxyl, and the introduction of fused-thiophene or -pyrrole on the pyrimidine core to strategically achieve conformational restriction. According to our biological evaluation, it was found that compound 9i could potently suppress EGFR^T790M/L858R kinase and H1975 cell proliferation, with IC₅₀ values of 4.902 nM and 0.6210 μM, respectively. Further study showed that 9i not only demonstrated highly selective inhibitory effects toward EGFR^T790M/L858R over wild-type EGFR (EGFR^WT), but it also had low cytotoxicity against normal HBE(human bronchial epithelial) and L-02 cells. Action mechanism studies showed that 9i effectively hindered cell migration and promoted apoptosis by AO(Acridine Orange)/EB(Ethidium Bromide) staining. These data would provide important clues for the screening of novel covalent EGFR^T790M/L858R inhibitors for non-small cell lung cancer (NSCLC) treatment.

设计并合成了三种类型的 2-arylamino-4-(piperidin-4-yloxy)pyrimidines (I-III) 作为共价 EGFR（表皮生长因子受体）^T790M/L858R抑制剂，取代了常见的 4-(3- amino)phenoxyl 部分与 4-(4-hydroxy)piperidine-4-oxyl，并在嘧啶核上引入稠合噻吩或 -pyrrole 以策略性地实现构象限制。根据我们的生物学评估，发现化合物9i可以有效抑制 EGFR ^T790M/L858R激酶和 H1975 细胞增殖，IC ₅₀值分别为 4.902 nM 和 0.6210 μM。进一步的研究表明，9i不仅对EGFR表现出高度选择性的抑制作用^T790M/L858R优于野生型 EGFR (EGFR ^WT )，但它对正常 HBE（人支气管上皮细胞）和 L-02 细胞的细胞毒性也较低。作用机制研究表明，通过 AO（吖啶橙）/EB（溴化乙锭）染色， 9i有效地阻碍细胞迁移并促进细胞凋亡。这些数据将为筛选用于非小细胞肺癌 (NSCLC) 治疗的新型共价 EGFR ^{T790M/L858R抑制剂提供重要线索。}