UNC51-like kinase1 (ULK1) recruits its binding partners and initiates the autophagy process in cancer. ULK1 is significantly overexpressed in Non-small cell lung cancer (NSCLC) and negatively correlated with clinical prognosis in NSCLC patients. Based upon the binding features of ULK1, we explored the pharmacophore modeling to discover the common anchoring features. It was verified by synthesizing 5-bromo-4-phenoxy-N-phenylpyrimidin-2-amine derivatives, as well as subsequently elucidating the structure-activity relationships (SAR). Among all the obtained ULK1 inhibitors, 5-bromo-4-(2-fluoro-4-nitrophenoxy)-N-(3,4,5-trimethoxyphenyl) pyrimidin-2-amine (3s), was the most active one. The docking analysis was conducted to compare 3s and SBI-0206965, which further elucidated the roles of the H-bond donor. This compound inhibited the proliferation of A549 cells and showed strong inhibitory activity against ULK1 kinase. Moreover, we found that compound 3s could induce apoptosis while simultaneously blocking autophagy. Collectively, these findings shed new light on compound 3s that would be utilized as a promising candidate drug for the future NSCLC therapy.

UNC51样激酶1（ULK1）募集其结合伴侣并启动癌症的自噬过程。ULK1在非小细胞肺癌（NSCLC）中明显过表达，并且与NSCLC患者的临床预后呈负相关。基于ULK1的结合特征，我们探索了药效团模型以发现常见的锚定特征。通过合成5-溴-4-苯氧基-N-苯基嘧啶-2-胺衍生物以及随后阐明结构-活性关系（SAR）进行了验证。在所有获得的ULK1抑制剂中，最活跃的是5-溴-4-（2-氟-4-硝基苯氧基）-N-（3,4,5-三甲氧基苯基）嘧啶-2-胺（3s）。进行对接分析以比较3s和SBI-0206965，它们进一步阐明了H键供体的作用。该化合物抑制A549细胞的增殖，并显示出对ULK1激酶的强抑制活性。此外，我们发现化合物3s可以诱导细胞凋亡，同时阻止自噬。总而言之，这些发现为化合物3s带来了新的亮点，该化合物将被用作未来NSCLC治疗的有希望的候选药物。