title: Ani9, A Novel Potent Small-Molecule ANO1 Inhibitor with Negligible Effect on ANO2.
abstract: Anoctamin1 (ANO1)/transmembrane protein 16A (TMEM16A), a calcium-activated chloride channel (CaCC), is involved in many physiological functions such as fluid secretion, smooth muscle contraction, nociception and cancer progression. To date, only a few ANO1 inhibitors have been described, and these have low potency and selectivity for ANO1. Here, we performed a high-throughput screening to identify highly potent and selective small molecule inhibitors of ANO1. Three novel ANO1 inhibitors were discovered from screening of 54,400 synthetic small molecules, and they were found to fully block ANO1 channel activity with an IC50 < 3 μM. Electrophysiological analysis revealed that the most potent inhibitor, 2-(4-chloro-2-methylphenoxy)-N-[(2-methoxyphenyl)methylideneamino]-acetamide (Ani9), completely inhibited ANO1 chloride current with submicromolar potency. Notably, unlike previous small-molecule ANO1 inhibitors identified to date, Ani9 displayed high selectivity for ANO1 as compared to ANO2, which shares a high amino acid homology to ANO1. In addition, Ani9 did not affect the intracellular calcium signaling and CFTR chloride channel activity. Our results suggest that Ani9 may be a useful pharmacological tool for studying ANO1 and a potential development candidate for drug therapy of cancer, hypertension, pain, diarrhea and asthma.
authors.affiliation: College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon 406-840, Korea.|Department of Integrated OMICS for Biomedical Science, WCU Program of Graduate School, Yonsei University, Seoul 120-749, Korea.
authors.forename: Yohan
authors.initials: Y
authors.lastname: Seo
authors.affiliation: Department of Integrated OMICS for Biomedical Science, WCU Program of Graduate School, Yonsei University, Seoul 120-749, Korea.
authors.forename: Ho K
authors.initials: HK
authors.lastname: Lee
authors.affiliation: College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon 406-840, Korea.|Department of Integrated OMICS for Biomedical Science, WCU Program of Graduate School, Yonsei University, Seoul 120-749, Korea.
authors.forename: Jinhong
authors.initials: J
authors.lastname: Park
authors.affiliation: College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon 406-840, Korea.
authors.forename: Dong-Kyu
authors.initials: DK
authors.lastname: Jeon
authors.affiliation: College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon 406-840, Korea.
authors.forename: Sungwoo
authors.initials: S
authors.lastname: Jo
authors.affiliation: College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon 406-840, Korea.
authors.forename: Minjae
authors.initials: M
authors.lastname: Jo
authors.affiliation: College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon 406-840, Korea.|Department of Integrated OMICS for Biomedical Science, WCU Program of Graduate School, Yonsei University, Seoul 120-749, Korea.
authors.forename: Wan
authors.initials: W
authors.lastname: Namkung
mesh_terms: D000081:Acetamides / Q000494:pharmacology*; D000075369:Anoctamin-1; D000075368:Anoctamins; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D018118:Chloride Channels / Q000037:antagonists & inhibitors* / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D057166:High-Throughput Screening Assays; D006801:Humans; D006835:Hydrazones / Q000494:pharmacology*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D054852:Small Molecule Libraries / Q000494:pharmacology*
chemical_list: C476213:ANO1 protein, human; C521849:ANO2 protein, human; D000081:Acetamides; C000621312:Ani9 compound; D000075369:Anoctamin-1; D000075368:Anoctamins; D018118:Chloride Channels; D006835:Hydrazones; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D054852:Small Molecule Libraries; D002118:Calcium