Mitoapocynin, a novel NOX2 inhibitor, mitigates nerve agents induced longterm neurotoxicity

Mitoapocynin 是一种新型 NOX2 抑制剂，可减轻神经毒剂引起的长期神经毒性

基本信息

批准号：
10178817
负责人：
Thimmasettappa Thippeswamy
金额：
$ 21.83万
依托单位：
IOWA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-06-15 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10178817
关键词：
Acute Address Adult Animal Model Animals Antiepileptogenic Atropine Behavioral Biological Assay Brain Brain Diseases Cause of Death Cessation of life Chemical Warfare Cholinesterase Inhibitors Cognitive Cues Devices Diazepam Disease Disease Progression Disease model Dose Electroencephalography England Epilepsy Epileptogenesis Exposure to Female Fostering Gliosis Health Histocytochemistry Hospitals Hour Human Impairment Implant Inflammatory Isoflurophate Legal patent Life Malaysia Midazolam Mission Modeling Modification Monitor Motor Motor Seizures Mus NADPH Oxidase Nerve Degeneration Neurologic Neuroprotective Agents Oral Organophosphates Oxidative Stress Oximes Parkinson Disease Pharmaceutical Preparations Populations at Risk Property Publications Rattus Recurrence Reporting Research Support Role Role Concepts Sarin Seizures Serum Soman Sprague-Dawley Rats Status Epilepticus Stress Survivors Symptoms Syria Telemetry Terrorism Testing Therapeutic Time Tokyo Wireless Technology behavior test blood-brain barrier permeabilization chemical threat cholinergic comorbidity conditioned fear cytokine effective therapy experimental study forced swim test improved in vivo in vivo evaluation inhibitor/antagonist kainate male medical countermeasure multiplex assay nerve agent neuroinflammation neuroprotection neurotoxicity novel prevent programs protective effect psychologic research and development toxic organophosphate insecticide exposure translational approach

项目摘要

Abstract Chemical warfare nerve agents (CWNA) are increasingly used to attack civilians worldwide. Currently, we lack effective treatment for CWNA survivors. Until recently, preventing acute death due to CWNA exposure had taken a top priority. However, reports on the life-long health consequences of sarin attack survivors are beginning to emerge. Like organophosphates (OPs), CWNAs are cholinesterase inhibitors and potent seizurogenic agents. In animal models, acute CWNA/OP exposure induces status epilepticus (SE) and other cholinergic symptoms. The current medical countermeasures (MCM) such as atropine, oxime, and diazepam/midazolam control symptoms, but do not prevent long-term neurotoxicity and comorbidity, primarily due to persistent nitrooxidative stress, neuroinflammation, and neurodegeneration. We predict that a combination of neuroprotectant and MCM can prevent CWNA-induced long-term neurotoxicity. We propose to investigate the long-term neuroprotective and antiepileptogenic effects of a novel inhibitor of NADPH oxidase (NOX2), mitoapocynin, in rat diisopropylfluorophosphate (DFP) and a nerve agent soman models. As proof of concept, we demonstrated the neuroprotective and disease-modifying properties of NOX2 inhibitors in the rat DFP and the mouse Parkinson's disease (PD) and seizure models. Treating mice or rats with mitoapocynin before the induction of SE with kainate (KA) or DFP significantly prevented the onset of convulsive seizures suggesting its anti-seizure property. We further tested mitoapocynin and diapocynin in telemetry-implanted rats after inducing SE with KA or DFP and treating them with MCM. Both NOX2 inhibitors are blood-brain barrier permeable. They prevented epileptogenesis in >50% rats and significantly reduced reactive gliosis and neurodegeneration. Although we had tested the proof of concept for the role of NOX2 in the rat DFP model, the optimum dose of diapocynin used was too high (300 mg/kg), which cannot be extrapolated for human use. Therefore, we developed, characterized, tested in vivo, and patented (US8962600 B2) the mitoapocynin, which is effective at 10-30 times lesser dose than diapocynin. Our overarching hypothesis is that mitoapocynin counteracts CWNA-induced long-term neurodegeneration, epileptogenesis, and restores brain function. We will investigate this in both male and female rat DFP models (Specific Aim 1) and validate in soman model (Specific Aim 2). We will expose rats to DFP/soman, treat with MCM, and 2h later with mitoapocynin (30 mg/kg, oral) or vehicle twice daily for the first three days, and 6 weeks later conduct a battery of behavioral tests. We will implant a telemetry device to monitor SRS and disease progression/modification in real-time. To determine the protective effect of mitoapocynin, we will conduct brain histochemistry for neurodegeneration and neuroinflammation, nitro-oxidative stress assays, and multiplex assay for cytokines. Our proposal embodies a novel translational approach targeting both neurodegeneration and epileptogenesis and may yield a novel orally-active therapy for CWNA-induced long-term neurotoxicity.

抽象的化学战神经毒剂（CWNA）越来越多地用于攻击全球平民。目前，我们缺乏 CWNA幸存者的有效治疗。直到最近，防止因CWNA暴露而导致的急性死亡优先。但是，关于沙林攻击幸存者终身健康后果的报告是开始出现。像有机磷酸盐（OPS）一样，CWNA是胆碱酯酶抑制剂和有效的蛋白质发育剂。在动物模型中，急性CWNA/OP暴露会诱导癫痫持续状态（SE）和其他胆碱能症状。当前的医疗对策（MCM），例如阿托品，氧赛和地西epa/咪达唑仑控制症状，但不能预防长期神经毒性和合并症，主要是由于持续的硝基氧化应激，神经炎症和神经变性。我们预测神经保护剂和MCM的组合可以防止CWNA诱导的长期神经毒性。我们建议研究新型NADPH抑制剂的长期神经保护和抗癫痫作用氧化酶（NOX2），线磷酸蛋白，在大鼠二异丙氟磷酸磷酸盐（DFP）和神经剂SOMAN模型中。作为概念证明，我们证明了NOX2抑制剂的神经保护和疾病改良特性在大鼠DFP和小鼠帕金森氏病（PD）和癫痫发作模型中。用在用海藻酸盐（KA）或DFP诱导SE之前，Mitoapocynin显着阻止了抽搐的发作癫痫发作暗示其抗塞氏菌财产。我们进一步测试了遥测植入式有线妇女素和肿瘤性的用KA或DFP诱导SE并用MCM对其进行治疗后，大鼠。两种NOX2抑制剂都是血脑屏障透水。他们阻止了> 50％大鼠的癫痫发生，并显着降低了反应性神经胶质变性和神经变性。尽管我们已经测试了NOX2在大鼠DFP模型中的作用的概念证明，但所用的共淋巴细胞蛋白的最佳剂量太高（300 mg/kg），不能推断人使用。因此，我们在体内开发，表征，测试，并获得专利（US8962600 B2）米托环蛋白，其剂量的剂量低于育呼吸蛋白的剂量10-30倍。我们的总体假设是线索蛋白抵消CWNA诱导的长期神经退行性，癫痫发生和恢复大脑功能。我们将在男性和女性大鼠DFP模型（特定目标1）中进行调查并验证在Soman模型中（特定目标2）。我们将使老鼠暴露于DFP/SOMAN，用MCM治疗，然后在2小时内与在头三天，每天两次Mitoapocynin（30 mg/kg，口服）或车辆，6周后进行电池行为测试。我们将植入遥测设备来监视SRS和疾病的进展/修饰实时。为了确定丝托辅酶的保护作用，我们将进行大脑组织化学神经变性和神经炎症，硝基氧化应激测定和细胞因子的多重测定。我们的建议体现了一种针对神经退行性和癫痫发生的新型翻译方法并可能产生一种新型的口腔活性疗法，用于CWNA诱导的长期神经毒性。