In this study, a novel class of thieno [2,3-d] pyrimidine derivatives containing resorcinol and morpholine fragments as Hsp90/mTOR dual inhibitors was designed, synthesized, and evaluated. In vitro anti-tumor assay results: the obtained compounds demonstrated effectiveness in suppressing the enzymatic activities of the Hsp90 and mTOR and inhibiting the proliferation of J82, T24, and SW780 cancer cell lines. Among these dual inhibitors, the most potent compound 17o, confirmed remarkable inhibitory activities on Hsp90, mTOR, and SW780 cell. Furthermore, the molecular dynamics simulation and a panel of mechanism studies revealed that inhibitor 17o suppressed the proliferation of SW780 cells through the over-activation of the PI3K/AKT/mTOR pathway regulated by mTOR inhibition and apoptosis regulated by the mitochondrial pathway. In subcutaneous J82 xenograft models, the compound 17o also presented considerable in vivo anti-tumor activity. Therefore, our investigations highlight that a new-found dual Hsp90/mTOR inhibitor by rational drug design strategies could be a promising lead compound for targeted bladder cancer therapy and deserves further studies.

在这项研究中，设计、合成和评估了一类新型噻吩并[2,3- d ]嘧啶衍生物，其中含有间苯二酚和吗啉片段作为 Hsp90/mTOR 双重抑制剂。体外抗肿瘤试验结果：获得的化合物在抑制 Hsp90 和 mTOR 的酶活性和抑制 J82、T24 和 SW780 癌细胞系的增殖方面具有有效性。在这些双重抑制剂中，最有效的化合物17o证实了对 Hsp90、mTOR 和 SW780 细胞的显着抑制活性。此外，分子动力学模拟和一组机制研究表明，抑制剂17o通过 mTOR 抑制调节的 PI3K/AKT/mTOR 通路的过度激活和线粒体通路调节的细胞凋亡来抑制 SW780 细胞的增殖。在皮下 J82 异种移植模型中，化合物17o也表现出相当大的体内抗肿瘤活性。因此，我们的研究强调，通过合理的药物设计策略新发现的双重 Hsp90/mTOR 抑制剂可能是靶向膀胱癌治疗的有前途的先导化合物，值得进一步研究。