In patients with non-small cell lung cancer (NSCLC), the standard therapy consists of selective tyrosine kinase inhibitors that target epidermal growth factor receptors (EGFR). Nonetheless, their clinical utility is primarily limited by the development of resistance to drugs. HDAC inhibitors have been shown in studies to reduce the level of EGFR that is expressed and downregulate the EGFR-induced phosphorylation of AKT and ERK. Therefore, dual inhibitors of EGFR and HDAC provide a potential approach as combination treatment synergistically inhibited the growth of NSCLC. Herein, we examined the EGFR inhibition effect of twenty compounds which designed and synthesized by us previously. Among them, compounds 12c and 12d exhibited powerful antiproliferative activity against the NCI–H1975 cell line with IC₅₀ values of 0.48 ± 0.07 and 0.35 ± 0.02 μM, correspondingly. In cell-free kinase assays, both 12c and 12d demonstrated target-specific EGFR inhibition against wild type (EGFR^wt). Furthermore, the expression of EGFR and phosphorylation of the EGF-induced pathways were significantly suppressed under the treatment of 12c and 12d. Besides, both histones H3 and H4 exhibited increased levels of acetylation following 12c and 12d treatment. The animal experiments shown that 12d could prevent the growth of tumor, inhibited the expression of EGFR and the phosphorylation levels of p70 S6K, AKT and p38 MAPK in vivo, and did not cause organ damage to the mice during the experiment.

Overall, the results illustrated that compound 12c and 12d could serve as effective EGFR and HDAC dual inhibitors in NSCLC cells. Our work offers an alternative strategy for NSCLC therapy.

对于非小细胞肺癌 (NSCLC) 患者，标准疗法包括针对表皮生长因子受体 (EGFR) 的选择性酪氨酸激酶抑制剂。尽管如此，它们的临床用途主要受到耐药性发展的限制。研究表明，HDAC 抑制剂可降低 EGFR 的表达水平，并下调 EGFR 诱导的 AKT 和 ERK 磷酸化。因此，EGFR和HDAC的双重抑制剂提供了一种潜在的方法，联合治疗可以协同抑制NSCLC的生长。在此，我们考察了我们之前设计合成的二十种化合物的EGFR抑制作用。其中，化合物12c和12d对NCI–H1975细胞系表现出强大的抗增殖活性，IC ₅₀值分别为0.48±0.07和0.35±0.02μM。在无细胞激酶测定中，12c和12d均表现出针对野生型 (EGFR ^wt )的靶标特异性 EGFR 抑制作用。此外，EGFR的表达和EGF诱导途径的磷酸化在12c和12d的处理下被显着抑制。此外，在12c和12d处理后，组蛋白 H3 和 H4 均表现出乙酰化水平增加。动物实验表明，12d可以阻止肿瘤的生长，抑制体内EGFR的表达以及p70 S6K、AKT和p38 MAPK的磷酸化水平，并且在实验过程中不会对小鼠造成器官损伤。

总体而言，结果表明化合物12c和12d可以作为NSCLC细胞中有效的EGFR和HDAC双重抑制剂。我们的工作为 NSCLC 治疗提供了另一种策略。