title: Treatment with afobazole at delayed time points following ischemic stroke improves long-term functional and histological outcomes.
abstract: There is currently a significant lack of therapeutic options for acute ischemic stroke, and no drug has been approved for treating patients at delayed time points (≥6h post-stroke). Afobazole, an anxiolytic currently used clinically in Russia, has been shown to reduce neuronal and glial cell injury in vitro following ischemia. Experiments using the permanent middle cerebral artery occlusion (MCAO) rat model were carried out to determine if afobazole can reduce ischemic stroke damage in vivo and expand the therapeutic window for stroke treatment. Post-stroke (24h) application of afobazole (0.3-3mg/kg) significantly decreased infarct volume at 96h post-surgery, as determined by Fluoro-Jade and NeuN staining of brain sections. Moreover, afobazole helped preserve both the levels and normal histological distribution of myelin basic protein, indicating a reduction in white matter injury. A time-dependence study showed that either pre-treatment or treatment started 6 to 48h post-stroke with the drug yields improved outcomes at 96h. The decrease in infarct volume produced by afobazole was blocked by the application of either a σ-1 (BD 1063, 30mg/kg) or a σ-2 (SM-21, 1mg/kg) antagonist, indicating that both receptor subtypes are involved in the effects of afobazole. Treatment with afobazole starting at 24h post-stroke resulted in enhanced survival one month following surgery. Behavioral testing of animals 28-32days post-surgery using the elevated body swing and forelimb grip-strength tests revealed that treatment with afobazole starting 24h post-stroke significantly reduces behavioral deficits caused by ischemic stroke. The increase in survival and improved functional outcomes are accompanied by a reduction in infarct volume, as determined by thionin staining of brain sections. Taken together, our data support the use of afobazole as a post-stroke pharmacological agent to expand the current therapeutic window.
authors.affiliation: Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida, 12901 Bruce B. Downs Blvd, MDC78, Tampa, FL 33612, USA.
authors.forename: C
authors.initials: C
authors.lastname: Katnik
authors.affiliation: Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida, 12901 Bruce B. Downs Blvd, MDC78, Tampa, FL 33612, USA.
authors.forename: A
authors.initials: A
authors.lastname: Garcia
authors.affiliation: Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida, 12901 Bruce B. Downs Blvd, MDC78, Tampa, FL 33612, USA.
authors.forename: A A
authors.initials: AA
authors.lastname: Behensky
authors.affiliation: IBC Generium, 27 Al. Soljenitsyna, Moscow 109004, Russia.
authors.forename: I E
authors.initials: IE
authors.lastname: Yasny
authors.affiliation: IBC Generium, 27 Al. Soljenitsyna, Moscow 109004, Russia.
authors.forename: A M
authors.initials: AM
authors.lastname: Shuster
authors.affiliation: State Foundation Institute of Pharmacology, Russian Academy of Medical Sciences, 8 Baltiyskaya, Moscow 125315, Russia.
authors.forename: S B
authors.initials: SB
authors.lastname: Seredenin
authors.affiliation: IBC Generium, 27 Al. Soljenitsyna, Moscow 109004, Russia.
authors.forename: A V
authors.initials: AV
authors.lastname: Petrov
authors.affiliation: Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida, 12901 Bruce B. Downs Blvd, MDC78, Tampa, FL 33612, USA.
authors.forename: S
authors.initials: S
authors.lastname: Seifu
authors.affiliation: Center for Excellence in Aging & Brain Repair, Morsani College of Medicine, USA; Department of Neurosurgery and Brain Repair, Morsani College of Medicine, USA.
authors.forename: J
authors.initials: J
authors.lastname: McAleer
authors.affiliation: Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida, 12901 Bruce B. Downs Blvd, MDC78, Tampa, FL 33612, USA; Center for Excellence in Aging & Brain Repair, Morsani College of Medicine, USA; Department of Neurosurgery and Brain Repair, Morsani College of Medicine, USA.
authors.forename: A
authors.initials: A
authors.lastname: Willing
authors.affiliation: Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida, 12901 Bruce B. Downs Blvd, MDC78, Tampa, FL 33612, USA. Electronic address: jcuevas@health.usf.edu.
authors.forename: J
authors.initials: J
authors.lastname: Cuevas
mesh_terms: D000818:Animals; D001562:Benzimidazoles / Q000008:administration & dosage / Q000627:therapeutic use*; D002545:Brain Ischemia / Q000188:drug therapy* / Q000473:pathology; D018737:Hand Strength / Q000502:physiology; D020244:Infarction, Middle Cerebral Artery / Q000188:drug therapy / Q000473:pathology; D008297:Male; D009025:Morpholines / Q000008:administration & dosage / Q000627:therapeutic use*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017480:Receptors, sigma / Q000819:agonists; D020521:Stroke / Q000188:drug therapy* / Q000473:pathology; D016896:Treatment Outcome
chemical_list: C410924:2-((2-morpholino)ethylthio)-5-ethoxybenzimidazole; D001562:Benzimidazoles; D009025:Morpholines; D017480:Receptors, sigma
keywords: Afobazole; Behavior; Ischemic stroke; Middle cerebral artery occlusion; Neuroprotection; Oligodendroglia; Sigma receptor