Identification and optimization of verapamil as a novel neuroprotective and anti-inflammatory agent for reducing long-term neurological morbidities following organophosphate-induced status epilepticus

维拉帕米作为新型神经保护和抗炎剂的鉴定和优化，用于减少有机磷引起的癫痫持续状态后的长期神经系统发病率

• 批准号: 10727765
• 负责人: Laxmikant S Deshpande
• 金额: $ 54.34万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10727765
• 关键词: Acetylcholinesterase Inhibitors; Acute; Alzheimer' s disease model; Anti-Inflammatory Agents; Antiinflammatory Effect; Anxiety; Atropine; Attenuated; Behavioral; Behavioral Assay; Brain; Brain Injuries; Brain region; Calcium Binding; Calcium Channel Blockers; Caring; Cell model; Cessation of life; Chemical Warfare; Chronic; Development; Dose; Drug Kinetics; Early Intervention; Electroencephalography; Emergency Care; Enzyme Inhibitor Drugs; Exhibits; Exposure to; Formulation; Glial Fibrillary Acidic Protein; Goals; Hour; Household; Hypertension; Immunohistochemistry; Impaired cognition; Industrialization; Inflammatory; Intoxication; Intramuscular; Intramuscular Injections; Ischemia; Mediating; Midazolam; Modeling; Morbidity - disease rate; Nerve Degeneration; Neurologic; Neurological outcome; Neuronal Injury; Neurons; Organophosphates; Outcome; Parkinson Disease; Pesticides; Pharmaceutical Preparations; Phase; Property; Rattus; Recurrence; Reducing Agents; Reporting; Research Support; Rodent; Role; Safety; Seizures; Solvents; Stains; Status Epilepticus; Stroke; Survivors; Techniques; Testing; Therapeutic; Toxic effect; United States National Institutes of Health; Up-Regulation; Verapamil; Water; central nervous system injury; chemical threat; cholinergic; cognitive performance; cytokine; effective therapy; field study; fluoro jade; fluorophosphate; functional outcomes; improved; in vivo; insight; ionization; mortality; mossy fiber; nerve agent; neuroinflammation; neuron loss; neuroprotection; novel; novel therapeutics; object recognition; pharmacologic; programs; standard of care; toxic organophosphate insecticide exposure; treatment duration

ABSTRACT Organophosphate (OP) compounds include pesticides and chemical warfare nerve agents. They are highly toxic and can produce a cholinergic crisis that rapidly progresses into status epilepticus (SE) and even death without emergency care. The standard-of-care (SOC) treatment with atropine, pralidoxime, and midazolam has dramatically improved survival after OP intoxication. Yet, many survivors of OP-SE exhibit brain injury, cognitive impairments, and spontaneous recurrent seizures (SRS). In addition, both acute and protracted neuro-inflammation and increased expression of pro-inflammatory cytokines have been reported following OP SE. These persistent neuroinflammatory changes are thought to underlie neurodegeneration, network hyper- excitability, and maladaptive plasticity, leading to cognitive dysfunction and SRS. Thus, mitigating neuro- inflammation is a primary target in alleviating neuronal injury and behavioral morbidities following OP SE. Verapamil (VPM) is a water-soluble calcium-channel blocker for high blood pressure and angina treatment. Recent findings have also demonstrated the potent neuroprotective and anti-inflammatory action of VPM in various CNS injuries. Our preliminary results showed that intramuscular VPM (10 mg/kg, i.m.) was safe and produced significant neuroprotection and decreased neuroinflammation in multiple brain regions when administered after the termination of DFP SE. It was also associated with decreased pro-inflammatory and upregulation of anti-inflammatory cytokines. Finally, this effect had a functional outcome since VPM improved anxiety and cognitive impairment at eight weeks post-DFP SE. Thus, this UG3-UH3 will investigate and optimize a VPM therapy as a potential countermeasure for treating OP SE that could be rapidly administered in the field. Studies will employ DFP-SE rat model to conduct the Specific Aims: In Aim 1, the safety of repeated i.m. injections will be assessed along with an assessment of pharmacokinetic parameters and the stability of VPM formulation. In Aim 2, the effects of VPM treatment on reducing neuronal death and neuroinflammation after DFP SE will be evaluated utilizing Fluoro-Jade C along with Glial Fibrillary Acidic Protein and Ionized calcium-binding adaptor molecule-1 immunohistochemical staining, respectively. We will also assess the effect of VPM therapy on pro- and anti-inflammatory cytokine expression. In Aim 3, VPM therapy will be optimized by studying the effects of various VPM doses and treatment durations on neurodegradation and neuroinflammation. In Aim 4, the functional outcomes of optimized VPM therapy on long-term anxiety, cognitive impairment, and SRS will be tested using a battery of rodent behavioral assays and EEG techniques. We will also draft a preliminary target product profile (TPP) by the end of our studies. These studies will provide further insight into the role of neuroinflammatory mechanisms in mediating OP morbidities and optimizing a VPM-based anti-inflammatory therapy as a novel countermeasure drug to alleviate OP toxicities.

抽象的 有机磷 (OP) 化合物包括杀虫剂和化学战神经毒剂。他们高度 有毒，可产生胆碱能危象，迅速发展为癫痫持续状态 (SE)，甚至死亡 无需紧急护理。阿托品、解磷定和咪达唑仑的标准护理 (SOC) 治疗 OP中毒后的生存率显着提高。然而，许多 OP-SE 幸存者表现出脑损伤， 认知障碍和自发性复发性癫痫发作（SRS）。此外，无论是急性的还是长期的 据报道，OP 后会出现神经炎症和促炎细胞因子表达增加 SE。这些持续的神经炎症变化被认为是神经退行性变、网络过度活跃的基础。 兴奋性和适应不良可塑性，导致认知功能障碍和 SRS。因此，减轻神经 炎症是减轻 OP SE 后神经元损伤和行为病态的主要目标。 维拉帕米 (VPM) 是一种水溶性钙通道阻滞剂，用于治疗高血压和心绞痛。 最近的研究结果还证明了 VPM 对神经保护和抗炎作用 各种中枢神经系统损伤。我们的初步结果表明，肌肉注射 VPM（10 mg/kg，肌肉注射）是安全且有效的。 产生显着的神经保护作用并减少多个大脑区域的神经炎症 DFP SE 终止后进行管理。它还与促炎性和促炎性降低有关 抗炎细胞因子的上调。最后，自从 VPM 改进以来，这种效果产生了功能性结果 DFP SE 后八周出现焦虑和认知障碍。因此，该 UG3-UH3 将调查并 优化 VPM 疗法作为治疗 OP SE 的潜在对策，可以在 领域。研究将采用 DFP-SE 大鼠模型来实现具体目标：在目标 1 中，重复实验的安全性 我是。将评估注射剂以及药代动力学参数和稳定性的评估 VPM 配方。在目标 2 中，VPM 治疗对减少神经元死亡和神经炎症的影响 在使用 Fluoro-Jade C 以及神经胶质纤维酸性蛋白和离子化蛋白对 DFP SE 进行评估后 分别进行钙结合接头分子-1免疫组织化学染色。我们也会评估效果 VPM 疗法对促炎和抗炎细胞因子表达的影响。在目标 3 中，VPM 疗法将通过以下方式进行优化： 研究不同 VPM 剂量和治疗持续时间对神经退化的影响 神经炎症。在目标 4 中，优化 VPM 疗法对长期焦虑的功能结果， 认知障碍和 SRS 将使用一系列啮齿动物行为测定和脑电图技术进行测试。 我们还将在研究结束时起草一份初步目标产品概况 (TPP)。这些研究将 进一步深入了解神经炎症机制在介导 OP 发病中的作用， 优化基于 VPM 的抗炎疗法作为减轻 OP 毒性的新型对策药物。