title: Screening of Transient Receptor Potential Canonical Channel Activators Identifies Novel Neurotrophic Piperazine Compounds.
abstract: Transient receptor potential canonical (TRPC) proteins form Ca(2+)-permeable cation channels activated upon stimulation of metabotropic receptors coupled to phospholipase C. Among the TRPC subfamily, TRPC3 and TRPC6 channels activated directly by diacylglycerol (DAG) play important roles in brain-derived neurotrophic factor (BDNF) signaling, promoting neuronal development and survival. In various disease models, BDNF restores neurologic deficits, but its therapeutic potential is limited by its poor pharmacokinetic profile. Elucidation of a framework for designing small molecules, which elicit BDNF-like activity via TRPC3 and TRPC6, establishes a solid basis to overcome this limitation. We discovered, through library screening, a group of piperazine-derived compounds that activate DAG-activated TRPC3/TRPC6/TRPC7 channels. The compounds [4-(5-chloro-2-methylphenyl)piperazin-1-yl](3-fluorophenyl)methanone (PPZ1) and 2-[4-(2,3-dimethylphenyl)piperazin-1-yl]-N-(2-ethoxyphenyl)acetamide (PPZ2) activated, in a dose-dependent manner, recombinant TRPC3/TRPC6/TRPC7 channels, but not other TRPCs, in human embryonic kidney cells. PPZ2 activated native TRPC6-like channels in smooth muscle cells isolated from rabbit portal vein. Also, PPZ2 evoked cation currents and Ca(2+) influx in rat cultured central neurons. Strikingly, both compounds induced BDNF-like neurite growth and neuroprotection, which were abolished by a knockdown or inhibition of TRPC3/TRPC6/TRPC7 in cultured neurons. Inhibitors of Ca(2+) signaling pathways, except calcineurin, impaired neurite outgrowth promotion induced by PPZ compounds. PPZ2 increased activation of the Ca(2+)-dependent transcription factor, cAMP response element-binding protein. These findings suggest that Ca(2+) signaling mediated by activation of DAG-activated TRPC channels underlies neurotrophic effects of PPZ compounds. Thus, piperazine-derived activators of DAG-activated TRPC channels provide important insights for future development of a new class of synthetic neurotrophic drugs.
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Seishiro
authors.initials: S
authors.lastname: Sawamura
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Masahiko
authors.initials: M
authors.lastname: Hatano
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Takada
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Kyosuke
authors.initials: K
authors.lastname: Hino
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Kawamura
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Jun
authors.initials: J
authors.lastname: Tanikawa
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nakagawa
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Hideharu
authors.initials: H
authors.lastname: Hase
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Akito
authors.initials: A
authors.lastname: Nakao
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Hirano
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Rachapun
authors.initials: R
authors.lastname: Rotrattanadumrong
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Shigeki
authors.initials: S
authors.lastname: Kiyonaka
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Masayuki X
authors.initials: MX
authors.lastname: Mori
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Motohiro
authors.initials: M
authors.lastname: Nishida
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Yaopeng
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Ryuji
authors.initials: R
authors.lastname: Inoue
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.).
authors.forename: Ryu
authors.initials: R
authors.lastname: Nagata
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering (S.S., Ma.H., Y.T., H.H., Mi.H., R.R., S.K., M.X.M., Y.M.), and Department of Technology and Ecology, Hall of Global Environmental Studies (S.K., Y.M.), Kyoto University, Kyoto, Japan; Sumitomo Dainippon Pharma Co., Ltd., Osaka, Japan (Y.T., K.H., T.K., J.T., H.N., R.N.); Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan (A.N.); Division of Cardiocirculatory Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, Okazaki, Japan (M.N.); and Department of Physiology, School of Medicine, Fukuoka University, Fukuoka, Japan (Y.H., R.I.) mori@sbchem.kyoto-u.ac.jp.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Mori
mesh_terms: D000818:Animals; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism; D009414:Nerve Growth Factors / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D010879:Piperazines / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D050052:TRPC Cation Channels / Q000819:agonists / Q000378:metabolism*
chemical_list: D009414:Nerve Growth Factors; D010879:Piperazines; D050052:TRPC Cation Channels