title: A novel multi-target ligand (JM-20) protects mitochondrial integrity, inhibits brain excitatory amino acid release and reduces cerebral ischemia injury in vitro and in vivo.
abstract: We previously showed that JM-20, a novel 1,5-benzodiazepine fused to a dihydropyridine moiety, possessed an anxiolytic profile similar to diazepam and strong neuroprotective activity in different cell models relevant to cerebral ischemia. Here, we investigated whether JM-20 protects against ischemic neuronal damage in vitro and in vivo. The effects of JM-20 were evaluated on hippocampal slices subjected to oxygen and glucose deprivation (OGD). For in vivo studies, Wistar rats were subjected 90 min of middle cerebral artery occlusion (MCAo) and oral administration of JM-20 at 2, 4 and 8 mg/kg 1 h following reperfusion. Twenty-four hours after cerebral blood flow restoration, neurological deficits were scored, and the infarct volume, histopathological changes in cortex, number of hippocampal and striatal neurons, and glutamate/aspartate concentrations in the cerebrospinal fluid were measured. Susceptibility to brain mitochondrial swelling, membrane potential dissipation, H2O2 generation, cytochrome c release, Ca2+ accumulation, and morphological changes in the organelles were assessed 24 h post-ischemia. In vitro, JM-20 (1 and 10 μM) administered during reperfusion significantly reduced cell death in hippocampal slices subjected to OGD. In vivo, JM-20 treatment (4 and 8 mg/kg) significantly decreased neurological deficit scores, edema formation, total infarct volumes and histological alterations in different brain regions. JM-20 treatment also protected brain mitochondria from ischemic damage, most likely by preventing Ca2+ accumulation in organelles. Moreover, an 8-mg/kg JM-20 dose reduced glutamate and aspartate concentrations in cerebrospinal fluid and the deleterious effects of MCAo even when delivered 8 h after blood flow restoration. These results suggest that in rats, JM-20 is a robust neuroprotective agent against ischemia/reperfusion injury with a wide therapeutic window. Our findings support the further examination of potential clinical JM-20 use to treat acute ischemic stroke.
authors: Centro de Investigación y Desarrollo de Medicamentos, Ave 26, No. 1605 Boyeros y Puentes Grandes, CP 10600, La Habana, Cuba.
authors: Yanier
authors: Y
authors: Nuñez-Figueredo
authors: Centro de Investigación y Desarrollo de Medicamentos, Ave 26, No. 1605 Boyeros y Puentes Grandes, CP 10600, La Habana, Cuba.
authors: Jeney
authors: J
authors: Ramírez-Sánchez
authors: Departamento de Bioquímica, PPG en Bioquímica, PPG en Educación en Ciencia, Instituto de Ciencias Básicas de la Salud, Universidad Federal de Rio Grande do Sul, Rua Ramiro Barcelos, 2600 Anexo, Porto Alegre, RS, 90035-003, Brazil.
authors: Gisele
authors: G
authors: Hansel
authors: Departamento de Bioquímica, PPG en Bioquímica, PPG en Educación en Ciencia, Instituto de Ciencias Básicas de la Salud, Universidad Federal de Rio Grande do Sul, Rua Ramiro Barcelos, 2600 Anexo, Porto Alegre, RS, 90035-003, Brazil.
authors: Elisa Nicoloso
authors: EN
authors: Simões Pires
authors: Centro de Investigación y Desarrollo de Medicamentos, Ave 26, No. 1605 Boyeros y Puentes Grandes, CP 10600, La Habana, Cuba.
authors: Nelson
authors: N
authors: Merino
authors: Centro de Investigación y Desarrollo de Medicamentos, Ave 26, No. 1605 Boyeros y Puentes Grandes, CP 10600, La Habana, Cuba.
authors: Odalys
authors: O
authors: Valdes
authors: Centro de Investigación y Desarrollo de Medicamentos, Ave 26, No. 1605 Boyeros y Puentes Grandes, CP 10600, La Habana, Cuba.
authors: René
authors: R
authors: Delgado-Hernández
authors: Centro de Investigación y Desarrollo de Medicamentos, Ave 26, No. 1605 Boyeros y Puentes Grandes, CP 10600, La Habana, Cuba.
authors: Alicia Lagarto
authors: AL
authors: Parra
authors: Laboratorio de Síntesis Orgánica de La Facultad de Química de La Universidad de La Habana (Zapata s/n entre G y Carlitos Aguirre, Vedado Plaza de la Revolución, CP 10400, La Habana, Cuba.
authors: Estael
authors: E
authors: Ochoa-Rodríguez
authors: Laboratorio de Síntesis Orgánica de La Facultad de Química de La Universidad de La Habana (Zapata s/n entre G y Carlitos Aguirre, Vedado Plaza de la Revolución, CP 10400, La Habana, Cuba.
authors: Yamila
authors: Y
authors: Verdecia-Reyes
authors: Departamento de Bioquímica, PPG en Bioquímica, PPG en Educación en Ciencia, Instituto de Ciencias Básicas de la Salud, Universidad Federal de Rio Grande do Sul, Rua Ramiro Barcelos, 2600 Anexo, Porto Alegre, RS, 90035-003, Brazil.
authors: Christianne
authors: C
authors: Salbego
authors: Laboratório de Neuroquímica y Biología Celular, Departamento de Biofunción / Bioquímica, Instituto de Ciencias de la Salud, Universidad Federal de Bahia, Av. Reitor Miguel Calmon s/n, Salvador, BA, 40.110-100, Brazil.
authors: Silvia L
authors: SL
authors: Costa
authors: Departamento de Bioquímica, PPG en Bioquímica, PPG en Educación en Ciencia, Instituto de Ciencias Básicas de la Salud, Universidad Federal de Rio Grande do Sul, Rua Ramiro Barcelos, 2600 Anexo, Porto Alegre, RS, 90035-003, Brazil.
authors: Diogo O
authors: DO
authors: Souza
authors: Centro de Estudio para las Investigaciones y Evaluaciones Biológicas, Instituto de Farmacia y Alimentos, Universidad de La Habana, Ave. 23 # 21425 e/214 y 222, La Coronela, La Lisa, CP 13600, La Habana, Cuba. Electronic address: gilbertopardo@infomed.sld.cu.
authors: Gilberto L
authors: GL
authors: Pardo-Andreu
mesh_terms: D000818:Animals; D001569:Benzodiazepines / Q000494:pharmacology*; D001921:Brain / Q000187:drug effects / Q000473:pathology / Q000503:physiopathology; D002545:Brain Ischemia / Q000188:drug therapy* / Q000473:pathology / Q000503:physiopathology; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D004195:Disease Models, Animal; D018846:Excitatory Amino Acids / Q000378:metabolism*; D005947:Glucose / Q000172:deficiency; D020244:Infarction, Middle Cerebral Artery / Q000188:drug therapy / Q000503:physiopathology; D008297:Male; D008928:Mitochondria / Q000187:drug effects* / Q000473:pathology / Q000502:physiology; D018696:Neuroprotective Agents / Q000494:pharmacology*; D009525:Niacin / Q000031:analogs & derivatives* / Q000494:pharmacology; D011897:Random Allocation; D017208:Rats, Wistar; D046509:Tissue Culture Techniques
chemical_list: C000591456:3-ethoxycarbonyl-2-methyl-4-(2-nitrophenyl)-4,11-dihydro-1H-pyrido(2,3-b)(1,5)benzodiazepine; D018846:Excitatory Amino Acids; D018696:Neuroprotective Agents; D001569:Benzodiazepines; D009525:Niacin; D005947:Glucose
keywords: Ischemia/reperfusion injury; JM-20; Middle cerebral artery occlusion; Mitochondria; Neuroprotection; Oxygen–glucose deprivation