Small‐molecule activators of TMEM16A, a calcium‐activated chloride channel, stimulate epithelial chloride secretion and intestinal contraction

TMEM16A (ANO1) is a calcium‐activated chloride channel (CaCC) expressed in secretory epithelia, smooth muscle, and other tissues. Cell‐based functional screening of ~110,000 compounds revealed compounds that activated TMEM16A CaCC conductance without increasing cytoplasmic Ca2+. By patch‐clamp, N‐aroylaminothiazole “activators” (Eact) strongly increased Cl‐ current at 0 Ca2+, whereas tetrazolylbenzamide “potentiators” (Fact) were not active at 0 Ca2+ but reduced the EC50 for Ca2+‐dependent TMEM16A activation. Of 682 analogs tested, the most potent activator (Eact) and potentiator (Fact) produced large and more sustained CaCC Cl‐currents than general agonists of Ca2+ signaling, with EC50 3–6 μM and Cl‐ conductance comparable to that induced transiently by Ca2+‐elevating purinergic agonists. Analogs of activators were identified that fully inhibited TMEM16A Cl‐ conductance, providing further evidence for direct TMEM16A binding. The TMEM16A activators increased CaCC conductance in human salivary and airway submucosal gland epithelial cells, and IL‐4 treated bronchial cells, and stimulated submucosal gland secretion in human bronchi and smooth muscle contraction in mouse intestine. Small‐molecule, TMEM16A‐targeted activators may be useful for drug therapy of cystic fibrosis, dry mouth, and gastrointestinal hypomotility disorders, and for pharmacological dissection of TMEM16A function.—Namkung, W., Yao, Z., Finkbeiner, W. E., Verkman, A. S. Small‐molecule activators of TMEM16A, a calcium‐activated chloride channel, stimulate epithelial chloride secretion and intestinal contraction. FASEB J. 25, 4048–4062 (2011). www.fasebj.org