Mitigation of Neurological Damage Following Seizures

项目概要-项目3 惊厥性化学威胁剂，如GABAA受体阻滞剂四亚甲基二磺基四胺（TETS） 和有机磷酸盐（OP）胆碱酯酶抑制剂二异丙基氟磷酸盐（DFP），可引发癫痫发作 发展为危及生命的癫痫持续状态（SE）。幸存者面临严重的长期发病率，包括 轻度至重度记忆丧失、情感障碍和反复发作（癫痫）。目前的医疗柜台- 措施不能充分防止这些神经缺陷。据推测，神经保护 将通过以下方式增强：（1）改善癫痫控制，和（2）将抗癫痫治疗与 减轻神经炎症和/或Ca 2+失调。在第一个项目期间，我们开发了一种鼠标， TETS诱导的SE模型，并改进了用于神经保护研究的DFP诱导的SE大鼠模型。使用 这些临床前模型，项目1和2表明，别孕烯醇酮，一种正变构调节剂， GABAA受体，是TETS诱导SE的上级对策，特别是当以 延迟时间别孕烯醇酮单独或与咪达唑仑组合不能终止DFP诱导的SE， 但是咪达唑仑、别孕烯醇酮和低剂量perampanel的联合肌内给药， 一种有效的AMPA受体拮抗剂，在终止DFP诱导的行为和 心电图SE，不引起明显镇静。我们还在候选神经- 暴露后用丹曲林（一种Ca 2+通道稳定剂）处理的保护剂，减少神经变性， 一种新的可溶性环氧化物水解酶（sEH）和环氧合酶-2（考克斯-2）的小分子双重抑制剂， 2)减弱小胶质细胞活化。核心A发现sEH抑制剂（sEHI）也使低血压正常化， 与大剂量地西泮或咪达唑仑有关在第二个项目期间，我们将量化以下方面的影响： 抗癫痫和神经保护治疗对神经退行性、神经炎症和功能结局的影响 使用体内成像纵向监测脑损伤与确证的组织学，24小时视频 脑电图，以量化复发性癫痫发作，以及认知，情感和 运动功能我们的目标是：（1）获得神经保护数据，以支持我们的抗癫痫电极的进展 别孕烯醇酮;（2）获得perampanel单独或与 与别孕烯醇酮组合;和（3）确定神经保护剂的线索，最初的重点是我们最 有前景的候选化合物：sEHI，双重sEH-COX-2抑制剂和丹曲林。我们将继续测试 项目2确定的其他抗癫痫候选药物和项目2确定的新型神经保护治疗药物 1使用体外模型。本项目中确定的上级神经保护治疗将由项目2进行测试 以评估它们是否与抗癫痫治疗相互作用。该项目的成果将支持 别孕烯醇酮对晚期发育的影响，并评估perampanel、其他候选抗癫痫药物 治疗或候选的神经保护治疗应该成为发展的主导。

Project Summary – Project 3 Convulsant chemical threat agents, such as the GABAA receptor blocker tetramethylenedisulfotetramine (TETS) and the organophosphate (OP) cholinesterase inhibitor diisopropylfluorophosphate (DFP), can trigger seizures that progress to life-threatening status epilepticus (SE). Survivors face significant, long-term morbidity, including mild-to-severe memory loss, affective disorders and recurrent seizures (epilepsy). Current medical counter- measures fail to sufficiently protect against these neurological deficits. It is hypothesized that neuroprotection will be enhanced by: (1) improving seizure control, and (2) combining antiseizure treatment with agents that mitigate neuroinflammation and/or Ca2+ dysregulation. During the first project period, we developed a mouse model of TETS-induced SE and refined a rat model of DFP-induced SE for use in neuroprotection studies. Using these preclinical models, Projects 1 and 2 showed that allopregnanolone, a positive allosteric modulator of GABAA receptors, was a superior countermeasure for TETS-induced SE, particularly when administered at delayed times. Allopregnanolone alone or in combination with midazolam did not terminate DFP-induced SE, but the combined intramuscular administration of midazolam, allopregnanolone and a low dose of perampanel, a potent AMPA receptor antagonist, was highly effective in terminating DFP-induced behavioral and electrographic SE without causing significant sedation. We also discovered in a screen of candidate neuro- protectants that post-exposure treatment with dantrolene, a Ca2+ channel stabilizer, reduced neurodegeneration, and that a novel small molecule dual inhibitor of soluble epoxide hydrolase (sEH) and cyclooxygenase-2 (COX- 2) attenuated microglial activation. Core A found that sEH inhibitors (sEHI) also normalized the hypotension associated with high-dose diazepam or midazolam. In the second project period, we will quantify the effects of antiseizure and neuroprotective treatments on neurodegenerative, neuroinflammatory and functional outcomes using in vivo imaging to longitudinally monitor brain damage with corroborative histology, 24 h video electroencephalography to quantify recurrent seizures, and behavioral assessments of cognitive, affective and motor function. Our goals are to: (1) obtain neuroprotection data to support the advance of our antiseizure lead allopregnanolone; (2) obtain proof-of-concept neuroprotective efficacy data for perampanel alone or in combination with allopregnanolone; and (3) identify neuroprotectant leads, with an initial focus on our most promising candidate compounds: sEHI, the dual sEH-COX-2 inhibitor and dantrolene. We will continue testing additional antiseizure candidates identified by Project 2 and novel neuroprotective treatments identified in Project 1 using in vitro models. Superior neuroprotective treatments identified in this project will be tested by Project 2 to assess whether they interact with antiseizure treatments. Results from this project will support transition of allopregnanolone to advanced development and assess whether perampanel, other candidate antiseizure treatments, or candidate neuroprotective treatments, should become development leads.