Key Laboratory of Natural Medicines of the Changbai Mountain, Ministry of Education, College of Pharmacy, Yanbian University, Yanji, Jilin Province, 133002, China.
ORxes Therapeutics (Shanghai）Co., Ltd., Shanghai 200120, China.
Bioorg Chem. 2024 Jul;148:107457. doi: 10.1016/j.bioorg.2024.107457. Epub 2024 May 15.
Based on the significant biological activities and the remarkable physical and chemical properties of 1H-1,2,3-triazole pharmacophore, we herein adopted the strategy of click chemistry to combine the triazole fragment and the unique scaffold of 25-OCH-PPD (AD-1) to design a series of potent compounds inducing apoptosis and DNA damage. The anti-proliferative effect was verified by MTT assay and colony formation assay. DNA double-stand breaks (DSBs) were obtained by observing the nuclear focus formation and the protein expression of γ-H2AX. Cell cycle arrest was evaluated by the cycle-related proteins such as CDK2, CDK4, CDK6, Cyclin D1 and P21. Apoptosis was assessed by flow cytometry, mitochondrial membrane potential (MMP) detection and the expression of apoptosis-related proteins. Reactive oxygen species (ROS) generation was measured with 2', 7'-dichlorofluorescein diacetate (DCFH-DA) staining. According to SAR analysis, the most potent compound 6a exhibited great inhibitory effect against A549 cells, which IC value of 2.84 ± 0.68 μM. Furthermore, 6a remarkably induced DNA damage, cell cycle arrest and apoptosis in A549 cells. 6a treatment increased the levels of ROS. Network pharmacology and molecular docking predicted the potential signaling pathways and ligand-receptor interactions, and the results of western blotting showed that 6a inhibited the PI3K/Akt/Bcl-2 signaling pathway by decreasing PI3K and Bcl-2 and total level of Akt expression, while Bax and Cyt c were increasing in 6a-treated A549 cells. As mentioned above, 6a has a potent inhibitory effect in A549 cells through induction of DNA damage, apoptosis via ROS generation and modulation of PI3K/Akt/Bcl-2 signaling pathway.
基于 1H-1,2,3-三唑药效团的显著生物学活性和优异的物理化学性质，我们采用点击化学策略，将三唑片段与 25-OCH-PPD（AD-1）的独特骨架相结合，设计了一系列能够诱导细胞凋亡和 DNA 损伤的有效化合物。通过 MTT 法和集落形成实验验证了抗增殖作用。通过观察核焦点形成和γ-H2AX 蛋白表达，获得 DNA 双链断裂（DSBs）。通过细胞周期相关蛋白如 CDK2、CDK4、CDK6、Cyclin D1 和 P21 评估细胞周期停滞。通过流式细胞术、线粒体膜电位（MMP）检测和凋亡相关蛋白的表达评估细胞凋亡。通过 2'，7'-二氯荧光素二乙酸酯（DCFH-DA）染色测量活性氧（ROS）的产生。根据 SAR 分析，最有效的化合物 6a 对 A549 细胞表现出很强的抑制作用，其 IC 值为 2.84±0.68μM。此外，6a 显著诱导 A549 细胞的 DNA 损伤、细胞周期停滞和凋亡。6a 处理增加了 ROS 水平。网络药理学和分子对接预测了潜在的信号通路和配体-受体相互作用，Western blot 结果表明，6a 通过降低 PI3K 和 Bcl-2 以及 Akt 总表达水平，抑制了 PI3K/Akt/Bcl-2 信号通路，而 Bax 和 Cyt c 在 6a 处理的 A549 细胞中增加。综上所述，6a 通过诱导 DNA 损伤、通过 ROS 生成和调节 PI3K/Akt/Bcl-2 信号通路来抑制 A549 细胞中的活性，从而具有很强的抑制作用。
