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

Heart Diseases
Fibrosis
Kidney
NFATC Transcription Factors
Gene Expression
Ureteral Obstruction
Muscle Cells
Biological Availability
Cations
Pharmacokinetics
BI 6
Pharmacology
Calcium
Pressure
Mutation
Growth
Therapeutics

Keywords

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

ASJC Scopus subject areas

  • General

Cite this

In vivo selective inhibition of TRPC6 by antagonist BI 749327 ameliorates fibrosis and dysfunction in cardiac and renal disease. / Lin, Brian Leei; Matera, Damian; Doerner, Julia F.; Zheng, Nan; Del Camino, Donato; Mishra, Sumita; Bian, Hong; Zeveleva, Svetlana; Zhen, Xiaoguang; Blair, Nathaniel T.; Chong, Jayhong A.; Hessler, David P.; Bedja, Djahida; Zhu, Guangshuo; Muller, Grace K.; Ranek, Mark; Pantages, Lynn; McFarland, Mary; Netherton, Matthew R.; Berry, Angela; Wong, Diane; Rast, Georg; Qian, Hu Sheng; Weldon, Steven M.; Kuo, Jay J.; Sauer, Achim; Sarko, Chris; Moran, Magdalene M.; Kass, David A; Pullen, Steven S.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, No. 20, 14.05.2019, p. 10156-10161.

Research output: Contribution to journalArticle

Lin, BL, Matera, D, Doerner, JF, Zheng, N, Del Camino, D, Mishra, S, Bian, H, Zeveleva, S, Zhen, X, Blair, NT, Chong, JA, Hessler, DP, Bedja, D, Zhu, G, Muller, GK, Ranek, M, Pantages, L, McFarland, M, Netherton, MR, Berry, A, Wong, D, Rast, G, Qian, HS, Weldon, SM, Kuo, JJ, Sauer, A, Sarko, C, Moran, MM, Kass, DA & Pullen, SS 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, vol. 116, no. 20, pp. 10156-10161. https://doi.org/10.1073/pnas.1815354116
Lin, Brian Leei ; Matera, Damian ; Doerner, Julia F. ; Zheng, Nan ; Del Camino, Donato ; Mishra, Sumita ; Bian, Hong ; Zeveleva, Svetlana ; Zhen, Xiaoguang ; Blair, Nathaniel T. ; Chong, Jayhong A. ; Hessler, David P. ; Bedja, Djahida ; Zhu, Guangshuo ; Muller, Grace K. ; Ranek, Mark ; Pantages, Lynn ; McFarland, Mary ; Netherton, Matthew R. ; Berry, Angela ; Wong, Diane ; Rast, Georg ; Qian, Hu Sheng ; Weldon, Steven M. ; Kuo, Jay J. ; Sauer, Achim ; Sarko, Chris ; Moran, Magdalene M. ; Kass, David A ; Pullen, Steven S. / In vivo selective inhibition of TRPC6 by antagonist BI 749327 ameliorates fibrosis and dysfunction in cardiac and renal disease. In: Proceedings of the National Academy of Sciences of the United States of America. 2019 ; Vol. 116, No. 20. pp. 10156-10161.
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AU - Lin, Brian Leei

AU - Matera, Damian

AU - Doerner, Julia F.

AU - Zheng, Nan

AU - Del Camino, Donato

AU - Mishra, Sumita

AU - Bian, Hong

AU - Zeveleva, Svetlana

AU - Zhen, Xiaoguang

AU - Blair, Nathaniel T.

AU - Chong, Jayhong A.

AU - Hessler, David P.

AU - Bedja, Djahida

AU - Zhu, Guangshuo

AU - Muller, Grace K.

AU - Ranek, Mark

AU - Pantages, Lynn

AU - McFarland, Mary

AU - Netherton, Matthew R.

AU - Berry, Angela

AU - Wong, Diane

AU - Rast, Georg

AU - Qian, Hu Sheng

AU - Weldon, Steven M.

AU - Kuo, Jay J.

AU - Sauer, Achim

AU - Sarko, Chris

AU - Moran, Magdalene M.

AU - Kass, David A

AU - Pullen, Steven S.

PY - 2019/5/14

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N2 - 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.

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KW - Fibrosis

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