The authors Alison J Davis, Jian Shi and Harry AT Pritchard contributed equally.
Potent vasorelaxant activity of the TMEM16A inhibitor T16Ainh-A01
Article first published online: 16 JAN 2013
© 2012 The Authors. British Journal of Pharmacology © 2012 The British Pharmacological Society
British Journal of Pharmacology
Volume 168, Issue 3, pages 773–784, February 2013
How to Cite
Davis, A. J., Shi, J., Pritchard, H. A., Chadha, P. S., Leblanc, N., Vasilikostas, G., Yao, Z., Verkman, A., Albert, A. P. and Greenwood, I. A. (2013), Potent vasorelaxant activity of the TMEM16A inhibitor T16Ainh-A01. British Journal of Pharmacology, 168: 773–784. doi: 10.1111/j.1476-5381.2012.02199.x
- Issue published online: 16 JAN 2013
- Article first published online: 16 JAN 2013
- Accepted manuscript online: 5 SEP 2012 05:03AM EST
- Manuscript Accepted: 15 AUG 2012
- Manuscript Revised: 25 JUL 2012
- Manuscript Received: 1 FEB 2012
- British Heart Foundation. Grant Numbers: PG/09/104, PG\07\127\24235, PG/08/042/2506, BB/J007226/1
- National Institutes of Health. Grant Numbers: 5 R01 HL 075477, 3 R01 HL075477-04S1
- calcium-activated chloride channels;
- vascular smooth muscle;
- patch clamp
Background and Purpose
T16Ainh-A01 is a recently identified inhibitor of the calcium-activated chloride channel TMEM16A. The aim of this study was to test the efficacy of T16Ainh-A01 for inhibition of calcium-activated chloride channels in vascular smooth muscle and consequent effects on vascular tone.
Single channel and whole cell patch clamp was performed on single smooth muscle cells from rabbit pulmonary artery and mouse thoracic aorta. Isometric tension studies were performed on mouse thoracic aorta and mesenteric artery as well as human abdominal visceral adipose artery.
In rabbit pulmonary artery myocytes T16Ainh-A01 (1–30 μM) inhibited single calcium (Ca2+)-activated chloride (Cl−) channels and whole cell currents activated by 500 nM free Ca2+. Similar effects were observed for single Ca2+-activated Cl− channels in mouse thoracic aorta, and in both cell types, channel activity was abolished by two antisera raised against TMEM16A but not by a bestrophin antibody. The TMEM16A potentiator, Fact (10 μM), increased single channel and whole cell Ca2+-activated Cl− currents in rabbit pulmonary arteries. In isometric tension studies, T16Ainh-A01 relaxed mouse thoracic aorta pre-contracted with methoxamine with an IC50 of 1.6 μM and suppressed the methoxamine concentration–effect curve. T16Ainh-A01 did not affect the maximal contraction produced by 60 mM KCl and the relaxant effect of 10 μM T16Ainh-A01 was not altered by incubation of mouse thoracic aorta in a cocktail of potassium (K+) channel blockers. T16Ainh-A01 (10 μM) also relaxed human visceral adipose arteries by 88 ± 3%.
Conclusions and Implications
T16Ainh-A01 blocks calcium-activated chloride channels in vascular smooth muscle cells and relaxes murine and human blood vessels.