Zheng Sheng, Shi Xiu-Yun, Su Xue, Wang Zhuo, Gao Hong-Wei, Wang Yu-Chen, Chen Xiang-Long, Zhang Wen, Zhang Hui
College of Life Science, Northwest Normal University, Lanzhou, Gansu 730030, PR China.
Laboratory Medicine Center, The Second Hospital & Clinical Medical School, Lanzhou University, Lanzhou, Gansu 730030, PR China.
Comput Methods Programs Biomed. 2025 Nov;271:108997. doi: 10.1016/j.cmpb.2025.108997. Epub 2025 Jul 28.
Agents targeting to Colchicine-site of tubulin have become a focus of attention in the development of new anti-cancer drugs due to their ability to overcome multi-drug resistant and anti-angiogenesis effects against tumor endothelium.
In this investigation, the machine learning methods and molecular docking approach were combined to identify promising destabilizing agents targeting for colchicine-site of tubulin.
Among these screened agents, hit22 represented excellent anti-tumor activity, which exhibited anti-proliferative activity toward H1299 cell with IC value of 3.93 μM, and significantly inhibited the colony formation in a dose-dependent manner. Furthermore, studies elucidated hit22 suppressed tubulin polymerization assembly in vitro, disrupted intracellular microtubule network, and caused G2/M phase cell cycle arrest and apoptosis in H1299 cells. Additionally, hit22 exhibited effective anti-migration abilities against H1299 cells, and suppressed angiogenesis of HUVECs. Importantly, hit22 could significantly inhibit tumor growth in the H1299 xenograft tumor model with a tumor growth inhibition rate of 70.30 %. The molecular dynamics simulations revealed hit22 could stably bind to colchicine site, interacting with some key residues, and inducing βT7 region to produce larger fluctuation. The average binding free energy for the Tubulin-hit22 was -90.9 kJ·mol.
Overall, these results illustrated that hit22 with novel scaffold might be a potential microtubule destabilizing agent targeting colchicine-site and deserves further investigation.
由于能够克服多药耐药性并对肿瘤内皮细胞产生抗血管生成作用，靶向微管蛋白秋水仙碱结合位点的药物已成为新型抗癌药物开发的关注焦点。
在本研究中，将机器学习方法与分子对接方法相结合，以鉴定有前景的靶向微管蛋白秋水仙碱结合位点的去稳定化药物。
在这些筛选出的药物中，hit22表现出优异的抗肿瘤活性，对H1299细胞具有抗增殖活性，IC值为3.93 μM，并以剂量依赖性方式显著抑制集落形成。此外，研究表明hit22在体外抑制微管蛋白聚合组装，破坏细胞内微管网络，并导致H1299细胞的G2/M期细胞周期阻滞和凋亡。此外，hit22对H1299细胞表现出有效的抗迁移能力，并抑制人脐静脉内皮细胞的血管生成。重要的是，hit22在H1299异种移植瘤模型中可显著抑制肿瘤生长，肿瘤生长抑制率为70.30%。分子动力学模拟显示hit22可稳定结合到秋水仙碱位点，与一些关键残基相互作用，并诱导βT7区域产生较大波动。微管蛋白-hit22的平均结合自由能为-90.9 kJ·mol。
总体而言，这些结果表明具有新型骨架的hit22可能是一种潜在的靶向秋水仙碱位点的微管去稳定化药物，值得进一步研究。
