title: A pharmacological screening approach for discovery of neuroprotective compounds in ischemic stroke.
abstract: With the availability and ease of small molecule production and design continuing to improve, robust, high-throughput methods for screening are increasingly necessary to find pharmacologically relevant compounds amongst the masses of potential candidates. Here, we demonstrate that a primary oxygen glucose deprivation assay in primary cortical neurons followed by secondary assays (i.e. post-treatment protocol in organotypic hippocampal slice cultures and cortical neurons) can be used as a robust screen to identify neuroprotective compounds with potential therapeutic efficacy. In our screen about 50% of the compounds in a library of pharmacologically active compounds displayed some degree of neuroprotective activity if tested in a pre-treatment toxicity assay but just a few of these compounds, including Carbenoxolone, remained active when tested in a post-treatment protocol. When further examined, Carbenoxolone also led to a significant reduction in infarction size and neuronal damage in the ischemic penumbra when administered six hours post middle cerebral artery occlusion in rats. Pharmacological testing of Carbenoxolone-related compounds, acting by inhibition of 11-β-hydroxysteroid dehydrogenase-1 (11β-HSD1), gave rise to similarly potent in vivo neuroprotection. This indicates that the increase of intracellular glucocorticoid levels mediated by 11β-HSD1 may be involved in the mechanism that exacerbates ischemic neuronal cell death, and inhibiting this enzyme could have potential therapeutic value for neuroprotective therapies in ischemic stroke and other neurodegenerative disorders associated with neuronal injury.
authors.affiliation: Behavioral and Functional Neuroscience Laboratory, Institute for Neuro-Innovation and Translational Neurosciences, School of Medicine, Stanford, California, USA.
authors.forename: Simret
authors.initials: S
authors.lastname: Beraki
authors.forename: Lily
authors.initials: L
authors.lastname: Litrus
authors.forename: Liza
authors.initials: L
authors.lastname: Soriano
authors.forename: Marie
authors.initials: M
authors.lastname: Monbureau
authors.forename: Lillian K
authors.initials: LK
authors.lastname: To
authors.forename: Steven P
authors.initials: SP
authors.lastname: Braithwaite
authors.forename: Karoly
authors.initials: K
authors.lastname: Nikolich
authors.forename: Roman
authors.initials: R
authors.lastname: Urfer
authors.forename: Donna
authors.initials: D
authors.lastname: Oksenberg
authors.forename: Mehrdad
authors.initials: M
authors.lastname: Shamloo
mesh_terms: D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1 / Q000037:antagonists & inhibitors; D000704:Analysis of Variance; D002545:Brain Ischemia / Q000188:drug therapy*; D002229:Carbenoxolone / Q000494:pharmacology; D055808:Drug Discovery / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005938:Glucocorticoids / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006624:Hippocampus / Q000166:cytology; D006801:Humans; D009474:Neurons / Q000187:drug effects; D018696:Neuroprotective Agents / Q000494:pharmacology*; D011419:Propidium; D018709:Statistics, Nonparametric
chemical_list: D005938:Glucocorticoids; D018696:Neuroprotective Agents; D011419:Propidium; D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1; D002229:Carbenoxolone