In vivo selective inhibition of TRPC6 by antagonist BI 749327 ameliorates fibrosis and dysfunction in cardiac and renal disease

Brian Leei Lin, Damian Matera, Julia F. Doerner, Nan Zheng, Donato Del Camino, Sumita Mishra, Hong Bian, Svetlana Zeveleva, Xiaoguang Zhen, Nathaniel T. Blair, Jayhong A. Chong, David P. Hessler, Djahida Bedja, Guangshuo Zhu, Grace K. Muller, Mark Ranek, Lynn Pantages, Mary McFarland, Matthew R. Netherton, Angela BerryDiane Wong, Georg Rast, Hu Sheng Qian, Steven M. Weldon, Jay J. Kuo, Achim Sauer, Chris Sarko, Magdalene M. Moran, David A Kass, Steven S. Pullen

Research output: Contribution to journalArticle

Abstract

Transient receptor potential canonical type 6 (TRPC6) is a nonselective receptor-operated cation channel that regulates reactive fibrosis and growth signaling. Increased TRPC6 activity from enhanced gene expression or gain-of-function mutations contribute to cardiac and/or renal disease. Despite evidence supporting a pathophysiological role, no orally bioavailable selective TRPC6 inhibitor has yet been developed and tested in vivo in disease models. Here, we report an orally bioavailable TRPC6 antagonist (BI 749327; IC 50 13 nM against mouse TRPC6, t 1/2 8.5–13.5 hours) with 85- and 42-fold selectivity over the most closely related channels, TRPC3 and TRPC7. TRPC6 calcium conductance results in the stimulation of nuclear factor of activated T cells (NFAT) that triggers pathological cardiac and renal fibrosis and disease. BI 749327 suppresses NFAT activation in HEK293T cells expressing wild-type or gain-of-function TRPC6 mutants (P112Q, M132T, R175Q, R895C, and R895L) and blocks associated signaling and expression of prohypertrophic genes in isolated myocytes. In vivo, BI 749327 (30 mg/kg/day, yielding unbound trough plasma concentration ∼180 nM) improves left heart function, reduces volume/mass ratio, and blunts expression of profibrotic genes and interstitial fibrosis in mice subjected to sustained pressure overload. Additionally, BI 749327 dose dependently reduces renal fibrosis and associated gene expression in mice with unilateral ureteral obstruction. These results provide in vivo evidence of therapeutic efficacy for a selective pharmacological TRPC6 inhibitor with oral bioavailability and suitable pharmacokinetics to ameliorate cardiac and renal stress-induced disease with fibrosis.

Original languageEnglish (US)
Pages (from-to)10156-10161
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume116
Issue number20
DOIs
StatePublished - May 14 2019

    Fingerprint

Keywords

  • Calcium
  • Fibrosis
  • Ion channels
  • Nuclear factor of activated T cells
  • TRPC6

ASJC Scopus subject areas

  • General

Cite this

Lin, B. L., Matera, D., Doerner, J. F., Zheng, N., Del Camino, D., Mishra, S., Bian, H., Zeveleva, S., Zhen, X., Blair, N. T., Chong, J. A., Hessler, D. P., Bedja, D., Zhu, G., Muller, G. K., Ranek, M., Pantages, L., McFarland, M., Netherton, M. R., ... Pullen, S. S. (2019). In vivo selective inhibition of TRPC6 by antagonist BI 749327 ameliorates fibrosis and dysfunction in cardiac and renal disease. Proceedings of the National Academy of Sciences of the United States of America, 116(20), 10156-10161. https://doi.org/10.1073/pnas.1815354116