Identification of novel TMEM16A blockers through integrated virtual screening, molecular dynamics, and experimental studies
Abstract 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. Thi...
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Nature Portfolio
2025-04-01
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| Online Access: | https://doi.org/10.1038/s41598-025-99751-w |
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| author | Mattanun Sangkhawasi Wichuda Pitaktrakul Rungtiwa Khumjiang Yasuteru Shigeta Chatchai Muanprasat Kowit Hengphasatporn Thanyada Rungrotmongkol |
| author_facet | Mattanun Sangkhawasi Wichuda Pitaktrakul Rungtiwa Khumjiang Yasuteru Shigeta Chatchai Muanprasat Kowit Hengphasatporn Thanyada Rungrotmongkol |
| author_sort | Mattanun Sangkhawasi |
| collection | DOAJ |
| description | Abstract 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 IC50 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. |
| format | Article |
| id | doaj-art-e1e7c46492ff45fd835ec58226e1e41e |
| institution | OA Journals |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Nature Portfolio |
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| spelling | doaj-art-e1e7c46492ff45fd835ec58226e1e41e2025-08-20T02:10:54ZengNature PortfolioScientific Reports2045-23222025-04-0115111110.1038/s41598-025-99751-wIdentification of novel TMEM16A blockers through integrated virtual screening, molecular dynamics, and experimental studiesMattanun Sangkhawasi0Wichuda Pitaktrakul1Rungtiwa Khumjiang2Yasuteru Shigeta3Chatchai Muanprasat4Kowit Hengphasatporn5Thanyada Rungrotmongkol6Center of Excellence in Biocatalyst and Sustainable Biotechnology, Department of Biochemistry, Faculty of Science, Chulalongkorn UniversityChakri Naruebodindra Medical Institute, Faculty of Medicine Ramathibodi Hospital, Mahidol UniversityChakri Naruebodindra Medical Institute, Faculty of Medicine Ramathibodi Hospital, Mahidol UniversityCenter for Computational Sciences, University of TsukubaChakri Naruebodindra Medical Institute, Faculty of Medicine Ramathibodi Hospital, Mahidol UniversityCenter for Computational Sciences, University of TsukubaCenter of Excellence in Biocatalyst and Sustainable Biotechnology, Department of Biochemistry, Faculty of Science, Chulalongkorn UniversityAbstract 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 IC50 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.https://doi.org/10.1038/s41598-025-99751-wTMEM16APore-blocker inhibitorVirtual screeningMolecular dynamicsCytotoxicityAnd electrophysiological assays |
| spellingShingle | Mattanun Sangkhawasi Wichuda Pitaktrakul Rungtiwa Khumjiang Yasuteru Shigeta Chatchai Muanprasat Kowit Hengphasatporn Thanyada Rungrotmongkol Identification of novel TMEM16A blockers through integrated virtual screening, molecular dynamics, and experimental studies Scientific Reports TMEM16A Pore-blocker inhibitor Virtual screening Molecular dynamics Cytotoxicity And electrophysiological assays |
| title | Identification of novel TMEM16A blockers through integrated virtual screening, molecular dynamics, and experimental studies |
| title_full | Identification of novel TMEM16A blockers through integrated virtual screening, molecular dynamics, and experimental studies |
| title_fullStr | Identification of novel TMEM16A blockers through integrated virtual screening, molecular dynamics, and experimental studies |
| title_full_unstemmed | Identification of novel TMEM16A blockers through integrated virtual screening, molecular dynamics, and experimental studies |
| title_short | Identification of novel TMEM16A blockers through integrated virtual screening, molecular dynamics, and experimental studies |
| title_sort | identification of novel tmem16a blockers through integrated virtual screening molecular dynamics and experimental studies |
| topic | TMEM16A Pore-blocker inhibitor Virtual screening Molecular dynamics Cytotoxicity And electrophysiological assays |
| url | https://doi.org/10.1038/s41598-025-99751-w |
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