Targeting EGFR and HER-2 is an essential direction for cancer treatment. Here, a series of N-(1,3,4-thiadiazol-2-yl)benzamide derivatives containing a 6,7-methoxyquinoline structure was designed and synthesized to serve as EGFR/HER-2 dual-target inhibitors. The kinase assays verified that target compounds could inhibit the kinase activity of EGFR and HER-2 selectively. The results of CCK-8 and 3D cell viability assays confirmed that target compounds had excellent anti-proliferation ability against breast cancer cells (MCF-7 and SK-BR-3) and lung cancer cells (A549 and H1975), particularly against SK-BR-3 cells, while the inhibitory effect on healthy breast cells (MCF-10A) and lung cells (Beas-2B) was weak. Among them, the hit compound YH-9 binded to EGFR and HER-2 stably in molecular dynamics studies. Further studies found that YH-9 could induce the release of cytochrome c and inhibit proliferation by promoting ROS expression in SK-BR-3 cells. Moreover, YH-9 could diminish the secretion of VEGF and bFGF factors in SK-BR-3 cells, then inhibited tube formation and angiogenesis. Notably, YH-9 could effectively inhibit breast cancer growth and angiogenesis with little toxicity in the SK-BR-3 cell xenograft model. Taken together, in vitro and in vivo results revealed that YH-9 had high drug potential as a dual-target inhibitor of EGFR/HER-2 to inhibit breast cancer growth and angiogenesis.

靶向EGFR和HER-2是癌症治疗的重要方向。在这里，设计并合成了一系列含有 6,7-甲氧基喹啉结构的N- (1,3,4-thiadiazol-2-yl)苯甲酰胺衍生物作为 EGFR/HER-2 双靶点抑制剂。激酶测定证实目标化合物可以选择性地抑制EGFR和HER-2的激酶活性。CCK-8 和 3D 细胞活力测定的结果证实，目标化合物对乳腺癌细胞（MCF-7 和 SK-BR-3）和肺癌细胞（A549 和 H1975）具有优异的抗增殖能力，尤其是对 SK- BR-3细胞，而对健康乳腺细胞（MCF-10A）和肺细胞（Beas-2B）的抑制作用较弱。其中，命中化合物YH-9在分子动力学研究中与 EGFR 和 HER-2 稳定结合。进一步研究发现，YH-9可以通过促进SK-BR-3细胞中ROS的表达来诱导细胞色素c的释放并抑制增殖。此外，YH-9可以减少 SK-BR-3 细胞中 VEGF 和 bFGF 因子的分泌，进而抑制管形成和血管生成。值得注意的是，在 SK-BR-3 细胞异种移植模型中，YH-9可以有效抑制乳腺癌的生长和血管生成，且毒性很小。总之，体外和体内结果表明，YH-9作为 EGFR/HER-2 的双靶点抑制剂具有很高的药物潜力，可抑制乳腺癌的生长和血管生成。