We designed 21 ethyl 3,5-diphenyl-2-cyclohexenone-6-carboxylate derivatives to identify compounds exhibiting anticancer activity. To measure the inhibitory effects of the compounds on cancer cell growth, a long-term survival clonogenic assay was performed. Since compounds containing a cyclohexenone moiety inhibit the enzyme acetylcholinesterase, an in vitro acetylcholinesterase assay was performed for all 21 cyclohexenone derivatives. To examine the effect of the derivative that exhibited the best cancer cell growth inhibition on the induction of apoptosis by demonstrating the activation of caspases and apoptosis regulatory proteins, immunoblotting and immunofluorescence microscopic analyses were performed. The binding mode between the cyclohexenone derivatives and acetylcholinesterase was elucidated at the molecular level using in silico docking. Druggability was evaluated based on ligand efficiency.

中文翻译：

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环己烯酮衍生物的抗癌活性

我们设计了21个3,5-二苯基-2-环己烯酮-6-羧酸乙酯衍生物，以鉴定具有抗癌活性的化合物。为了测量化合物对癌细胞生长的抑制作用，进行了长期存活克隆形成测定。由于含有环己烯酮部分的化合物会抑制乙酰胆碱酯酶，因此对所有21种环己烯酮衍生物进行了体外乙酰胆碱酯酶测定。为了通过证明胱天蛋白酶和凋亡调节蛋白的活化来检查表现出最佳的癌细胞生长抑制作用的衍生物对凋亡诱导的作用，进行了免疫印迹和免疫荧光显微镜分析。使用计算机对接在分子水平上阐明了环己烯酮衍生物与乙酰胆碱酯酶之间的结合方式。