Screening in a cell-based assay for inhibitors of microglial nitric oxide production reveals calmodulin-regulated protein kinases as potential drug discovery targets.

Détails

ID Serval
serval:BIB_8B3A9F819CAE
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Screening in a cell-based assay for inhibitors of microglial nitric oxide production reveals calmodulin-regulated protein kinases as potential drug discovery targets.
Périodique
Brain Research
Auteur(s)
Mirzoeva S., Koppal T., Petrova T.V., Lukas T.J., Watterson D.M., Van Eldik L.J.
ISSN
0006-8993 (Print)
ISSN-L
0006-8993
Statut éditorial
Publié
Date de publication
1999
Volume
844
Numéro
1-2
Pages
126-134
Langue
anglais
Résumé
A high-throughput screening (HTS) assay for inhibitors of nitric oxide (NO) production by activated microglia was developed and used to compare the relative activities of various anti-inflammatory compounds and cell-permeable protein kinase inhibitors. BV-2 cells, an immortalized line that retains phenotypic features of microglia and produces NO in response to lipopolysaccharide (LPS), were used in the activation paradigm for the HTS assay. A characteristic feature of the compounds that were the most potent dose-dependent inhibitors of NO production is their ability to modulate serine/threonine protein kinases. The anti-inflammatory compound K252a, an inhibitor of calmodulin (CaM)-regulated protein kinases, had one of the highest potencies in the assay. Other classes of kinase inhibitors, including the protein kinase A inhibitor H-89, the mitogen activated protein kinase inhibitors PD98059 and SB203580, and the tyrosine kinase inhibitor genistein, were less potent and efficacious than K252a or the general serine/threonine/tyrosine kinase inhibitor staurosporine. K252a suppresses production of the inducible nitric-oxide synthase (iNOS). The inhibitory effect of K252a is not due to cell toxicity and does not correlate with inhibition of NFkappaB nuclear translocation. The mechanism of action appears to involve inhibition of phosphorylation of the transcription factor CREB, a protein whose activity is modulated by phosphorylation by CaM-dependent protein kinases. These data suggest that signal transduction pathways mediated by CaM-dependent protein kinases warrant future study as potential drug discovery targets.
Mots-clé
Amidines/pharmacology, Animals, Benzylamines/pharmacology, Calcium Signaling/drug effects, Calcium-Calmodulin-Dependent Protein Kinases/metabolism, Carbazoles/pharmacology, Cell Line, Cell Nucleus/metabolism, Cyclic AMP Response Element-Binding Protein/metabolism, Drug Design, Enzyme Inhibitors/pharmacology, Flavonoids/pharmacology, Imidazoles/pharmacology, Indole Alkaloids, Isoquinolines/pharmacology, Lipopolysaccharides/pharmacology, Mice, Microglia/cytology, Microglia/enzymology, NF-kappa B/metabolism, Nitric Oxide/metabolism, Nitric Oxide Synthase/antagonists & inhibitors, Nitric Oxide Synthase/metabolism, Phosphorylation, Pyridines/pharmacology, Staurosporine/pharmacology, Sulfonamides
Pubmed
Web of science
Création de la notice
20/12/2012 16:52
Dernière modification de la notice
03/03/2018 19:11
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