As a class of microtubule targeting agents, colchicine binding site inhibitors (CBSIs) are considered as promising drug candidates for cancer therapy. However, due to adverse reactions, there are currently no CBSIs approved by FDA for cancer treatment. Therefore, extensive efforts are still encouraged to find novel CBSIs with different chemical structures and better anticancer efficacies. In this work, we designed and synthesized a new coumarin-dihydroquinoxalone derivative, MY-673, and evaluated its anticancer potency in vitro and in vivo. We confirmed that MY-673 was a potent CBSI that it not only inhibited tubulin polymerization, but also exhibited significant inhibitory potency on the growth of 13 cancer cells with IC₅₀ values from 11.7 nM to 395.9 nM. Based on the results of kinase panel screening, MY-673 could inhibit ERK (extracellular regulated protein kinases) pathways-related kinases. We further confirmed that MY-673 could inhibit ERK signaling pathway in MGC-803 and HGC-27 cells, and then affected the expression level of SMAD4 protein in TGF-β (transforming growth factor β) /SMAD (small mother against decapentaplegic) signaling pathway using the western blotting assay. In addition, compound MY-673 could effectively inhibit cell proliferation, migration and induce cell apoptosis. We also further confirmed the in vivo efficacy of MY-673 in inhibiting tumor growth using the MGC-803 xenograft tumor model. At 20 mg/kg, the TGI rate was 85.9%, and it did not cause obvious toxicity to the main organs of mice. Together, the results we report here indicated that MY-673 was a promising CBSI for cancer treatment, which was capable of inhibiting the ERK pathway with potent antiproliferative activities in vitro and in vivo.

作为一类微管靶向剂，秋水仙碱结合位点抑制剂 (CBSI) 被认为是有前途的癌症治疗候选药物。然而，由于不良反应，目前还没有FDA批准用于癌症治疗的CBSI。因此，仍然鼓励广泛努力寻找具有不同化学结构和更好抗癌功效的新型CBSI。在这项工作中，我们设计并合成了一种新的香豆素-二氢喹恶酮衍生物MY-673，并评估了其体外和体内的抗癌效力。我们证实MY-673是一种有效的 CBSI，它不仅抑制微管蛋白聚合，而且对具有 IC 的 13 种癌细胞的生长表现出显着的抑制效力_{从 11.7 nM 到 395.9 nM 的50 个}值。根据激酶面板筛选的结果，MY-673可以抑制 ERK（细胞外调节蛋白激酶）通路相关的激酶。我们进一步证实MY-673可以抑制MGC-803和HGC-27细胞中的ERK信号通路，进而影响TGF-β（转化生长因子β）/SMAD（小母抗脑瘫）信号通路中SMAD4蛋白的表达水平使用蛋白质印迹分析的途径。此外，复方MY-673能有效抑制细胞增殖、迁移，诱导细胞凋亡。我们还进一步证实了MY-673的体内功效使用 MGC-803 异种移植肿瘤模型抑制肿瘤生长。20 mg/kg时，TGI率为85.9%，对小鼠主要脏器无明显毒性。总之，我们在此报告的结果表明，MY-673是一种很有前途的癌症治疗 CBSI，它能够抑制 ERK 通路，在体外和体内具有强大的抗增殖活性。