Chen Zenan, Li Wei, Zhao Yan, Liu Dingrui, Han Jiahong, Cai Enbo
College of Chinese Medicinal Material, Jilin Agricultural University, 2888 Xincheng Street, Changchun 130118, China.
Molecules. 2025 Apr 8;30(8):1667. doi: 10.3390/molecules30081667.
Tumorigenesis and progression are closely associated with apoptosis and primarily regulated by mitochondria, which are considered major targets for cancer therapy. In this study, twelve novel icaritin (ICT) derivatives were designed and synthesized, four of which were specifically targeted to mitochondria. Biological studies demonstrated that all compounds containing triphenylphosphine (TPP) exhibited a substantial increase in antitumor activity compared to ICT and control compounds while also exhibiting notable selectivity for tumor cells over normal cells. Among these derivatives, Mito-ICT-4 exhibited the strongest antiproliferative effect, with an IC value of 0.73 ± 0.06 μM for BEL-7402 cells, which is 29 times lower than that of ICT, and an IC value of 67.11 ± 2.09 μM for HEK293 cells, indicating approximately 33-fold selectivity for tumor cells. High-performance liquid chromatography (HPLC) analysis revealed that Mito-ICT-4 significantly accumulated in the mitochondria of BEL-7402 cells, with the level of accumulation approximately 2.5 times greater than that of ICT. Further investigations demonstrated that upon entering the mitochondria of tumor cells, Mito-ICT-4 downregulated SIRT3 protein expression, disrupted intracellular redox homeostasis, and led to a substantial increase in mitochondrial ROS levels, abnormal CypD-dependent MPTP opening, mitochondrial membrane potential depolarization, and ROS release into the cytoplasm, ultimately triggering ROS-mediated apoptosis in BEL-7402 cells. Transcriptomic analysis identified differentially expressed genes and enriched pathways, highlighting the ROS-mediated p38-MAPK signaling pathway as a key mediator of Mito-ICT-4-induced mitochondria-dependent apoptosis. The effects of Mito-ICT-4 on the expression of key genes (SIRT3, CypD, P-MKK6, P-P38, and DDIT3) were further validated by qRT-PCR and Western blot analysis, with results aligning with transcriptomic data. The novel ICT derivatives synthesized in this study, with mitochondria-targeting functionality, provide a basis for the development of targeted antitumor drugs.
肿瘤发生和进展与细胞凋亡密切相关，主要由线粒体调控，线粒体被认为是癌症治疗的主要靶点。在本研究中，设计并合成了12种新型淫羊藿素（ICT）衍生物，其中4种特异性靶向线粒体。生物学研究表明，与ICT和对照化合物相比，所有含三苯基膦（TPP）的化合物抗肿瘤活性均显著增强，同时对肿瘤细胞表现出显著高于正常细胞的选择性。在这些衍生物中，Mito-ICT-4表现出最强的抗增殖作用，对BEL-7402细胞的IC值为0.73±0.06μM，比ICT低29倍，对HEK293细胞的IC值为67.11±2.09μM，表明对肿瘤细胞具有约33倍的选择性。高效液相色谱（HPLC）分析显示Mito-ICT-4在BEL-7402细胞线粒体中显著蓄积，蓄积水平约为ICT的2.5倍。进一步研究表明，Mito-ICT-4进入肿瘤细胞线粒体后下调SIRT3蛋白表达，破坏细胞内氧化还原稳态，导致线粒体ROS水平大幅升高、CypD依赖性MPTP异常开放、线粒体膜电位去极化以及ROS释放到细胞质中，最终触发BEL-7402细胞中ROS介导的凋亡。转录组分析鉴定了差异表达基因并富集了相关通路，突出了ROS介导的p38-MAPK信号通路作为Mito-ICT-4诱导的线粒体依赖性凋亡的关键介质。通过qRT-PCR和蛋白质免疫印迹分析进一步验证了Mito-ICT-4对关键基因（SIRT3、CypD、P-MKK6、P-P38和DDIT3）表达的影响，结果与转录组数据一致。本研究合成的具有线粒体靶向功能的新型ICT衍生物为开发靶向抗肿瘤药物提供了依据。
