A novel dual NO-donating oxime and c-Jun N-terminal kinase inhibitor protects against cerebral ischemia–reperfusion injury in mice

Abstract

The c-Jun N-terminal kinase (JNK) has been shown to be an important regulator of neuronal cell death. Previously, we synthesized the sodium salt of 11H-indeno[1,2-b]quinoxalin-11-one (IQ-1S) and demonstrated that it was a high-affinity inhibitor of the JNK family. In the present work, we found that IQ-1S could release nitric oxide (NO) during its enzymatic metabolism by liver microsomes. Moreover, serum nitrite/nitrate concentration in mice increased after intraperitoneal injection of IQ-1S. Because of these dual actions as JNK inhibitor and NO-donor, the therapeutic potential of IQ-1S was evaluated in an animal stroke model. We subjected wild-type C57BL6 mice to focal ischemia (30 minutes) with subsequent reperfusion (48 hours). Mice were treated with IQ-1S (25 mg/kg) suspended in 10% solutol or with vehicle alone 30 minutes before and 24 hours after middle cerebral artery MCA) occlusion (MCAO). Using laser-Doppler flowmetry, we monitored cerebral blood flow (CBF) above the MCA during 30 minutes of MCAO provoked by a filament and during the first 30 minutes of subsequent reperfusion. In mice treated with IQ-1S, ischemic and reperfusion values of CBF were not different from vehicle-treated mice. However, IQ-1S treated mice demonstrated markedly reduced neurological deficit and infarct volumes as compared with vehicle-treated mice after 48 hours of reperfusion. Our results indicate that the novel JNK inhibitor releases NO during its oxidoreductive bioconversion and improves stroke outcome in a mouse model of cerebral reperfusion. We conclude that IQ-1S is a promising dual functional agent for the treatment of cerebral ischemia and reperfusion injury.

JNK-IN-8, a c-Jun N-terminal kinase inhibitor, improves functional recovery through suppressing neuroinflammation in ischemic stroke

“…c-Jun N-terminal kinases (JNK) are important stress responsive kinases, and JNK signaling is the shared pathway linking microglia activation, neuroinflammation and cerebral ischemic injury [18, 19]. Previous studies demonstrated that JNK inhibition by specific inhibitor (e.g., IQ-1S and SP600125) contributes to reduce cerebral ischemia-reperfusion injury in mice [20, 21]. JNK-IN-8, an adenosine triphosphate-competitive irreversible pan-JNK inhibitor, exerts a protective role in inhibiting traumatic brain injury and cancer cell grow [22, 23].…”

“…Another study demonstrated that SP600125 attenuated subarachnoid hemorrhage-cerebral vasospasm through a suppressed inflammatory response [33]. IQ-1S releases NO during its oxidoreductive bioconversion and improves stroke outcome in a mouse model of cerebral reperfusion [20]. Noteworthily, different JNK inhibitor exerts diverse physiological properties because of targeting different members of the JNK family.…”

“…A specific JNK inhibitor, JNK-IN-8, is a potent and selective covalent inhibitor of JNK and has been used to investigate the precise role of JNK in many pathways and diseases [9]. In a previous study, a dual NO-donating oxime and c-Jun N-terminal kinase inhibitor is reported to protect cells against cerebral ischemia-reperfusion injury in mice, indicating the use of JNK inhibitor is accessible for investigation on functions of JNK in ischemia injury [10]. Specifically, JNK-IN-8 is reported to significantly suppress tumor growth in vitro and in vivo, directly proving JNK regulating breast cancer tumorigenesis [11].…”

Targeting thrombomodulin to circulating red blood cells augments its protective effects in models of endotoxemia and ischemia‐reperfusion injury

“…At the end of the reperfusion period, mice were examined for neurologic deficits using a 4‐point scale before being humanely killed. Neurologic outcomes were graded as: 0, normal; 1, forepaw monoparesis; 2, circling to right; 3, falling to right; and 4, no spontaneous walking and depressed consciousness (45). The brain was cut into 2 mm thick coronal blocks and stained in the dark with 2, 3, 5‐triphenyltetrazolium chloride for 1 h at 37°C, and the infarct volume was calculated (46).…”

Ischemic stroke and myocardial infarction: promising therapeutic goals for protecting the brain and heart

“…Сегодня следует учитывать частое сочетание кардиальной патологии у пациентов с инсультом, поэтому оценка фармакологической модуляция JNK-активности является актуальной как для первичной, так и вторичной профилактики сердечнососудистых и цереброваскулярных заболеваний. Перспективность нейропротекторных и кардиопротекторных препаратов уже доказана, поскольку контроль JNK-активности происходит на уровне фосфорилирования и дефосфорилирования [6,7,8,11], а препараты, которые являются NO-донорским оксимом и ингибитором JNK, проявляют двойные эффекты, способствующие нейропротекции [11]. Точные молекулярные механизмы модуляции передачи сигналов JNK при ишемии или реперфузии остаются сегодня неясными для многих агентов, демонстрирующих нейропротекторный и кардиопротекторный эффект, что открывает путь для создания новых терапевтических подходов с безопасным профилем эффективности при одновременном сочетании острой и хронической патологии головного мозга и сердца [9,10,11].…”