Abstract
DY-9760e, 3-[2-[4-(3-chloro-2-methylphenyl)-1-piperazinyl]ethyl]-5, 6-dimethoxy-1-(4-imidazolylmethyl)-1H-indazole dihydrochloride 3.5 hydrate, a novel calmodulin (CaM) antagonist, possesses neuroprotective activity. In the current study, we examined the effects of DY-9760e on nitric oxide synthase (NOS) activities in vitro and on calcium ionophore-induced NO production in situ. DY-9760e inhibited both neuronal NOS and endothelial NOS activities without affecting inducible NOS activity. It also inhibited purified neuronal NOS activity with a potency similar to that seen for purified CaM kinase II activity in vitro. Furthermore, DY-9760e significantly inhibited Ca(2+) ionophore (A23187)-induced NO production in mouse N1E-115 neuroblastoma cells, at a concentration of less than 1 microM. In contrast, no apparent inhibitory effect on Ca(2+)/CaM-dependent protein kinase II activity was observed in cultured hippocampal neurons up to 5 microM. These results suggest that the inhibitory effect of DY-9760e on CaM-dependent NOS activities underlies neuroprotective effects of the agent.
MeSH terms
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Analysis of Variance
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Animals
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Calcium-Calmodulin-Dependent Protein Kinase Type 2
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Calcium-Calmodulin-Dependent Protein Kinases / antagonists & inhibitors
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Calmodulin / antagonists & inhibitors
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Cell Extracts / analysis
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Enzyme Inhibitors / pharmacology*
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Hippocampus / drug effects
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Hippocampus / enzymology
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Indazoles / pharmacology*
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Neuroblastoma
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Neurons / drug effects*
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Neurons / enzymology
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Neuroprotective Agents / pharmacology*
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Nitric Oxide / metabolism
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Nitric Oxide Synthase / antagonists & inhibitors*
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Nitric Oxide Synthase Type I
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Nitric Oxide Synthase Type II
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Nitric Oxide Synthase Type III
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Rats
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Sulfonamides / pharmacology
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Tumor Cells, Cultured
Substances
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3-(2-(4-(3-chloro-2-methylphenyl)1-piperazinyl)ethyl)5,6-dimethoxy-1-(4-imidazolylmethyl)-1H-indazol dihydrochloride 3.5 hydrate
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Calmodulin
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Cell Extracts
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Enzyme Inhibitors
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Indazoles
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Neuroprotective Agents
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Sulfonamides
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Nitric Oxide
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W 7
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Nitric Oxide Synthase
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Nitric Oxide Synthase Type I
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Nitric Oxide Synthase Type II
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Nitric Oxide Synthase Type III
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Nos1 protein, rat
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Nos2 protein, rat
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Nos3 protein, rat
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Calcium-Calmodulin-Dependent Protein Kinase Type 2
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Calcium-Calmodulin-Dependent Protein Kinases