A series of novel tetrahydrobenzothieno[2,3-d]pyrimidine urea derivatives was synthesized according to fragment-based design strategy. They were evaluated for their anticancer activity against MCF-7 cell line. Three compounds 9c, 9d and 11b showed 1.5–1.03 folds more potent anticancer activity than doxorubicin. In this study, a promising multi-sited enzyme small molecule inhibitor 9c, which showed the most potent anti-proliferative activity, was identified. The anti-proliferative activity of this compound appears to correlate well with its ability to inhibit topoisomerase II (IC₅₀ = 9.29 μM). Moreover, compound 9c showed excellent VEGFR-2 inhibitory activity, at the sub-micromolar level with IC₅₀ value 0.2 μM, which is 2.1 folds more potent than sorafenib. Moreover, activation of damage response pathway of the DNA leads to cell cycle arrest at G2/M phase, accumulation of cells in pre-G1 phase and annexin-V and propidium iodide staining, indicating that cell death proceeds through an apoptotic mechanism. Compound 9c showed potent pro-apoptotic effect through induction of the intrinsic mitochondrial pathway of apoptosis. This mechanistic pathway was confirmed by a significant increase in the expression of the tumor suppressor gene p53, elevation in Bax/BCL-2 ratio and a significant increase in the level of active caspase-3. Quantitative structure-activity relationship (QSAR) studies delivered equations of five 3D descriptors with R² = 0.814. This QSAR model provides an effective technique for understanding the observed antitumor properties and thus could be adopted for developing effective lead structures.

根据基于片段的设计策略，合成了一系列新颖的四氢苯并噻吩并[2,3- d ]嘧啶脲衍生物。评价它们对MCF-7细胞系的抗癌活性。三种化合物9c，9d和11b的有效抗癌活性比阿霉素高1.5-1.03倍。在这项研究中，鉴定出了显示出最有效的抗增殖活性的有前途的多部位酶小分子抑制剂9c。该化合物的抗增殖活性似乎与其抑制拓扑异构酶II的能力密切相关（IC ₅₀  = 9.29μM）。此外，化合物9c在亚微摩尔水平显示出优异的VEGFR-2抑制活性，IC ₅₀值为0.2μM，效力比索拉非尼高2.1倍。此外，DNA损伤反应途径的激活导致细胞周期停滞在G2 / M期，G1期前细胞蓄积，膜联蛋白V和碘化丙锭染色，表明细胞死亡是通过凋亡机制进行的。化合物9c通过诱导内在的线粒体凋亡途径，显示出有效的促凋亡作用。肿瘤抑制基因p53的表达显着增加，Bax / BCL-2比的升高以及活性caspase-3的水平显着增加，证实了这种机制途径。定量构效关系（QSAR）研究提供了5个3D描述符的方程，R ²  = 0.814。该QSAR模型提供了一种有效的技术，可用于了解观察到的抗肿瘤特性，因此可用于开发有效的铅结构。