Novel Anticonvulsant and Neuroprotective Therapies for TETS and OP Intoxication

加州大学戴维斯分校反ACT卓越中心的主要目标是识别和推进改进 用于快速终止癫痫发作和减轻迟发性神经学后果的医学对策 是由于急性中毒引起的该中心包括三个研究 项目：项目1通过体外机制筛选发现治疗候选物， 抗癫痫和神经保护作用的项目2和3分别。这些项目依靠三个科学的 支持药物分析和生物标志物检测的核心（核心A），药物化学和药理学 测试（核心B），以及实验设计和数据分析（核心C）。研究教育核心支持 对策研究培训，行政核心监督和协调科学和 行政业务。该中心专注于GABAA受体拮抗剂四亚甲基二磺基四胺 （TETS）和有机磷酸胆碱酯酶抑制剂二异丙基氟磷酸盐（DFP），其可以触发 发展为危及生命的癫痫持续状态（SE）的惊厥。幸存者面临着重大的，长期的 发病率，包括轻度至重度记忆丧失、情感障碍和复发性癫痫。当前医学 对抗措施可以降低暴露个体的死亡率，但它们这样做有显著的副作用， 只有在暴露后几分钟内给药才能发挥最大的作用。这些局限性突出表明， 需要改进医疗对策。在第一个项目期间，我们开发了创新的体外 用于基于机制的筛选以鉴定候选抗癫痫发作和神经保护治疗剂的平台， 以及重现在人类中观察到的急性癫痫发作活动和神经缺陷的新型临床前模型 TETS或OP急性中毒后死亡利用这些模型，我们发现：（1）别孕烯醇酮， GABAA受体阳性变构调节剂，是TETS诱导SE的一种上级对策，和（2） 将标准治疗与别孕烯醇酮和低剂量perampanel（一种强效AMPA受体）联合使用 拮抗剂在终止DFP诱导的SE方面比单独的标准治疗更有效。我们还发现 暴露后用丹曲林（一种Ca2+通道稳定剂）或 新型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。我们 在这个第二个项目期间的目标是：（1）推进我们的抗癫痫铅别孕烯醇酮;（2）继续 开发别孕烯醇酮和perampanel;（3）确定辅助神经保护电极， 双重sEH-COX-2抑制剂和丹曲林;（4）进行机制研究，以发现新的 治疗候选人。我们在第二个项目期间的里程碑是：（一）提供数据和监管 发展用于治疗GABAAR拮抗剂诱导癫痫发作的别孕烯醇酮的策略;（ii）确定 别孕烯醇酮和perampanel联合治疗是否值得作为先导药物开发 "通用解毒剂";以及（iii）确定改善长期结果的主要神经保护治疗。

The primary objective of the UC Davis CounterACT Center of Excellence is to identify and advance improved medical countermeasures for rapidly terminating seizures and mitigating the delayed neurologic consequences following acute intoxication with convulsant chemical threat agents. The Center comprises three research projects: Project 1 discovers therapeutic candidates via in vitro mechanistic screens, which are tested for in vivo antiseizure and neuroprotective efficacy by Projects 2 and 3, respectively. The projects rely on three scientific cores to support drug analysis and biomarker detection (Core A), medicinal chemistry and pharmacological testing (Core B), and experimental design and data analysis (Core C). A Research Education Core supports training in countermeasure research, and an Administrative Core oversees and coordinates scientific and administrative operations. The Center focuses on the GABAA receptor antagonist tetramethylenedisulfotetramine (TETS) and the organophosphate cholinesterase inhibitor diisopropylfluorophosphate (DFP), which can trigger convulsions 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. Current medical countermeasures can reduce mortality in exposed individuals, but they do so with significant side effects and are maximally effective only if administered within minutes of exposure. These limitations underscore the urgent need for improved medical countermeasures. In the first project period, we developed innovative in vitro platforms for mechanism-based screening to identify candidate antiseizure and neuroprotective therapeutics, and novel preclinical models that recapitulate acute seizure activity and neurological deficits observed in humans following acute intoxication with TETS or OPs. Using these models, we discovered: (1) allopregnanolone, a GABAA receptor positive allosteric modulator, was a superior countermeasure for TETS-induced SE, and (2) combining standard-of-care with allopregnanolone and a low dose of perampanel, a potent AMPA receptor antagonist, was more effective than standard-of-care alone in terminating DFP-induced SE. We also discovered that neuropathology was mitigated by post-exposure treatment with dantrolene, a Ca2+ channel stabilizer, or a novel small molecule dual inhibitor of soluble epoxide hydrolase (sEH) and cyclooxygenase-2 (COX-2). Our goals in this second project period are to: (1) advance our antiseizure lead allopregnanolone; (2) continue development of allopregnanolone and perampanel; (3) identify adjunct neuroprotective leads, focusing initially on the dual sEH-COX-2 inhibitor and dantrolene; and (4) conduct mechanistic studies to discover new therapeutic candidates. Our milestones for the second project period are to: (i) produce data and a regulatory strategy to advance allopregnanolone for treatment of GABAAR antagonist-induced seizures; (ii) determine whether combination treatment with allopregnanolone and perampanel warrants development as a lead “universal antidote”; and (iii) identify lead neuroprotective treatments for improving long-term outcomes.