Fan Guangrui, Luo Xiongfei, Shi Yibo, Wang Yingru, Ji Luhua, Gong Yuwen, Yang Engaung, Chen Chaohu, Cui Shu, Ding Hui, Zhang Zhijun, Wang Juan, Liu Yingqian, Wang Zhiping
Institute of Urology, Lanzhou University Second Hospital, Urinary System Disease Clinical Medical Research Center of Gansu Province, NO. 82 Cuiying Gate, Lanzhou 730000, Gansu Province, China.
School of Pharmacy, Lanzhou University, No. 199, Donggang West Road, Lanzhou 730000, Gansu Province, China.
Bioorg Chem. 2024 Dec;153:107802. doi: 10.1016/j.bioorg.2024.107802. Epub 2024 Sep 5.
The treatment of bladder cancer is limited by low drug efficacy and drug resistance. Hence, this study aimed to screen and identify potential drug precursors and investigate their mechanism of action. A set of camptothecin derivatives showing high anti-tumor potential was selected from early-stage research or literature and synthesized to construct a compound library. A total of 135 compounds were screened in T24 and J82 cells, revealing that FL118 significantly inhibited the proliferation of GC (gemcitabine + cisplatin)-sensitive/insensitive cells. FL118 exhibited excellent penetration and killing ability in organoids and three GC-insensitive patient-derived xenografts. Chemical proteomic and docking calculations were employed to identify binding proteins, indicating that FL118 can bind into H2A.X and its entwined DNA. The results of Cellular thermal shift assay and surface plasmon resonance (K = 3.77E-6) support the above findings. Fluorescence localization revealed widespread binding of FL118 within the cell nucleus. Furthermore, WB showed that FL118 increased cellular DNA damage, resulting in significant cell cycle inhibition. The binding of FL118 to H2A.X hindered the damage repair process, leading to apoptosis. Controllable adverse reactions were observed in mice treated with FL118. In conclusion, FL118 may be a superior anti-bladder cancer compound that acts as a molecular glue binding to both H2A.X and DNA. The resistance mediated by the DNA damage repair to DNA damage caused by GC regimen can be reversed by FL118. This distinct mechanism of FL118 has the potential to complement existing mainstream treatment approaches for bladder cancer.
膀胱癌的治疗受到药物疗效低和耐药性的限制。因此，本研究旨在筛选和鉴定潜在的药物前体，并研究其作用机制。从早期研究或文献中选择了一组具有高抗肿瘤潜力的喜树碱衍生物并进行合成，以构建一个化合物库。在T24和J82细胞中总共筛选了135种化合物，结果表明FL118显著抑制了吉西他滨+顺铂（GC）敏感/不敏感细胞的增殖。FL118在类器官和三种GC不敏感的患者来源异种移植模型中表现出优异的渗透和杀伤能力。采用化学蛋白质组学和对接计算来鉴定结合蛋白，表明FL118可以与H2A.X及其缠绕的DNA结合。细胞热位移分析和表面等离子体共振（K = 3.77E-6）的结果支持上述发现。荧光定位显示FL118在细胞核内广泛结合。此外，蛋白质免疫印迹显示FL118增加了细胞DNA损伤，导致显著的细胞周期抑制。FL118与H2A.X的结合阻碍了损伤修复过程，导致细胞凋亡。在用FL118治疗的小鼠中观察到可控的不良反应。总之，FL118可能是一种优异的抗膀胱癌化合物，它作为一种分子胶水与H2A.X和DNA结合。FL118可以逆转由DNA损伤修复介导的对GC方案引起的DNA损伤的耐药性。FL118这种独特的作用机制有可能补充现有的膀胱癌主流治疗方法。
