@article{1ec6f814fc6b4a16acc1b012703ae292,
title = "In vivo selective inhibition of TRPC6 by antagonist BI 749327 ameliorates fibrosis and dysfunction in cardiac and renal disease",
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; IC50 13 nM against mouse TRPC6, t1/2 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.",
keywords = "Calcium, Fibrosis, Ion channels, Nuclear factor of activated T cells, TRPC6",
author = "Lin, {Brian Leei} and Damian Matera and Doerner, {Julia F.} and Nan Zheng and {Del Camino}, Donato and Sumita Mishra and Hong Bian and Svetlana Zeveleva and Xiaoguang Zhen and Blair, {Nathaniel T.} and Chong, {Jayhong A.} and Hessler, {David P.} and Djahida Bedja and Guangshuo Zhu and Muller, {Grace K.} and Ranek, {Mark J.} and Lynn Pantages and Mary McFarland and Netherton, {Matthew R.} and Angela Berry and Diane Wong and Georg Rast and Qian, {Hu Sheng} and Weldon, {Steven M.} and Kuo, {Jay J.} and Achim Sauer and Chris Sarko and Moran, {Magdalene M.} and Kass, {David A.} and Pullen, {Steven S.}",
note = "Funding Information: ACKNOWLEDGMENTS. We acknowledge the contributions of Glenn Gibson, Kathleen Lincoln, and Valentina Berger toward the in vivo renal pharmacology studies. This work was supported by Boehringer Ingelheim and NIH Grants R01-HL-119012, HL-131358, R35-HL135827, and PO-HL107153 (to D.A.K.), T32 Grant T32-HL-7227 (to B.L.L.), and American Heart Association Postdoctoral Fellowship Grants 18CDA34110140 (to M.J.R.) and 18POST33960157 (to G.K.M.). Funding Information: We acknowledge the contributions of Glenn Gibson, Kathleen Lincoln, and Valentina Berger toward the in vivo renal pharmacology studies. This work was supported by Boehringer Ingelheim and NIH Grants R01-HL-119012, HL-131358, R35-HL135827, and PO-HL107153 (to D.A.K.), T32 Grant T32-HL-7227 (to B.L.L.), and American Heart Association Postdoctoral Fellowship Grants 18CDA34110140 (to M.J.R.) and 18POST33960157 (to G.K.M.). Funding Information: Author contributions: B.L.L., D.M., J.F.D., D.d.C., D.W., J.J.K., A.S., C.S., M.M.M., D.A.K., and S.S.P. designed research; B.L.L., D.M., J.F.D., N.Z., D.d.C., S.M., H.B., S.Z., X.Z., N.T.B., J.A.C., D.P.H., D.B., G.Z., G.K.M., L.P., and M.M. performed research; M.R.N., A.B., and C.S. contributed new reagents/analytic tools; B.L.L., D.M., J.F.D., N.Z., D.d.C., S.M., H.B., S.Z., X.Z., N.T.B., J.A.C., D.P.H., D.B., G.K.M., M.J.R., D.W., G.R., H.S.Q., S.M.W., J.J.K., A.S., M.M.M., D.A.K., and S.S.P. analyzed data; and B.L.L., G.R., A.S., D.A.K., and S.S.P. wrote the paper. Conflict of interest statement: D.M., J.F.D., L.P., D.W., G.R., S.M.W., S.Z., H.S.Q, J.J.K, A.S., and S.S.P. are full-time employees of Boehringer Ingelheim Pharmaceuticals, Inc. M.M, M.R.N, A.B., and C.S. were full-time employees of Boehringer Ingelheim Pharmaceuticals, Inc. A.B. and M.R.N. are listed as coinventors on a US provisional patent application filed by Boehringer Ingelheim Pharmaceuticals, Inc. relevant to this work. J.F.D., N.Z., D.d.C., X.Z., N.T.B., J.A.C., D.P.H., and M.M.M. were employees of Hydra Biosciences, and received options. This work was supported in part by Boehringer Ingelheim Pharmaceuticals, Inc. This article is a PNAS Direct Submission. Published under the PNAS license. 1To whom correspondence may be addressed. Email: dkass@jhmi.edu or steven.pullen@ boehringer-ingelheim.com. This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. 1073/pnas.1815354116/-/DCSupplemental. Published online April 26, 2019. Publisher Copyright: {\textcopyright} 2019 National Academy of Sciences. All rights reserved. Copyright: Copyright 2019 Elsevier B.V., All rights reserved.",
year = "2019",
month = may,
day = "14",
doi = "10.1073/pnas.1815354116",
language = "English (US)",
volume = "116",
pages = "10156--10161",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "National Academy of Sciences",
number = "20",
}