Sangkhawasi Mattanun, Pitaktrakul Wichuda, Khumjiang Rungtiwa, Shigeta Yasuteru, Muanprasat Chatchai, Hengphasatporn Kowit, Rungrotmongkol Thanyada
Center of Excellence in Biocatalyst and Sustainable Biotechnology, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand.
Chakri Naruebodindra Medical Institute, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bang Pla, Bang Pli, Samut Prakan, 10540, Thailand.
Sci Rep. 2025 Apr 29;15(1):15065. doi: 10.1038/s41598-025-99751-w.
The calcium-activated chloride channel TMEM16A is a promising drug target for treating hypertension, secretory diarrheas, and various cancers, including head and neck cancer. Despite its potential, no FDA-approved drugs have provided the structural basis for directly inhibiting TMEM16A. This study aims to identify a novel pore-blocker of TMEM16A by integrating virtual screening, molecular dynamics simulations, and in vitro studies. Using the calcium-bound structure of TMEM16A with and without the pore-blocker 1PBC, we performed virtual screening on nearly 90,000 compounds from the ChemDiv database. Approximately 67% of these compounds demonstrated better binding affinity than 1PBC. Among the top 20 compounds selected for short-circuit current assays using human lung adenocarcinoma cells (Calu-3), compounds N066-0059, N066-0060, and N066-0067 inhibited TMEM16A activity with IC values of 0.24 µM, 0.41 µM, and 0.48 µM, respectively, which was lower than that of a positive control Ani9 (9 µM). Due to its highest potency in electrophysiological assays, N066-0059 was subjected to mechanistic studies. Molecular dynamics simulations elucidated its binding stability and strength, showing superior performance to 1PBC over 500 ns with 3 replicates. This study advances TMEM16A-targeted drug development, offering new insights for anticancer therapies.
钙激活氯离子通道TMEM16A是治疗高血压、分泌性腹泻以及包括头颈癌在内的多种癌症的一个很有前景的药物靶点。尽管其具有潜力，但尚无美国食品药品监督管理局（FDA）批准的药物提供直接抑制TMEM16A的结构基础。本研究旨在通过整合虚拟筛选、分子动力学模拟和体外研究来鉴定一种新型的TMEM16A孔道阻滞剂。利用有和没有孔道阻滞剂1PBC的TMEM16A钙结合结构，我们对来自ChemDiv数据库的近90,000种化合物进行了虚拟筛选。这些化合物中约67%表现出比1PBC更好的结合亲和力。在选择用于使用人肺腺癌细胞（Calu-3）进行短路电流测定的前20种化合物中，化合物N066-0059、N066-0060和N066-0067抑制TMEM16A活性的IC值分别为0.24 µM、0.41 µM和0.48 µM，低于阳性对照Ani9（9 µM）。由于其在电生理测定中的效力最高，N066-0059被用于机制研究。分子动力学模拟阐明了其结合稳定性和强度，在500纳秒内进行3次重复显示其性能优于1PBC。本研究推动了以TMEM16A为靶点的药物开发，为抗癌治疗提供了新的见解。
