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.

摘要 有机磷酸盐（OP）化合物包括杀虫剂和化学战神经毒剂。它们是高度 有毒，并可产生胆碱能危象，迅速发展为癫痫持续状态（SE），甚至死亡 没有紧急护理。阿托品、解磷定和咪达唑仑的标准治疗（SOC） 大大提高了OP中毒后的存活率。然而，许多OP-SE的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.这些持续的神经炎性变化被认为是神经变性、网络超敏反应、 兴奋性和适应不良的可塑性，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发病率的主要目标。 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。 最近的研究结果也证明了VPM在神经系统疾病中的有效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，i.m.）安全且 在多个脑区产生显著的神经保护和减少神经炎症， 在DFP SE终止后给予。它还与减少促炎性和 抗炎细胞因子的上调。最后，由于VPM改善，该效应具有功能性结局 DFP SE后8周的焦虑和认知障碍。因此，UG 3-UH 3将进行调查， 优化VPM治疗，作为治疗OP SE的潜在对策，可在 外地研究将采用DFP-SE大鼠模型进行具体目的：在目的1中， I.M.将沿着对药物动力学参数和药物稳定性的评估， VPM制剂。在目的2中，VPM治疗对减少神经元死亡和神经炎症的作用 DFP后，将使用Fluoro-Jade C沿着胶质纤维酸性蛋白和离子化 钙结合衔接分子-1免疫组织化学染色。我们也会评估 VPM治疗对促炎和抗炎细胞因子表达的影响。在目标3中，将通过以下方式优化VPM治疗： 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治疗对长期焦虑的功能结局， 认知障碍和SRS将使用一系列啮齿动物行为测定和EEG技术进行测试。 我们还将在研究结束时起草初步目标产品简介（TPP）。这些研究将 进一步了解神经炎症机制在介导OP发病率中的作用， 优化基于VPM的抗炎疗法作为减轻OP毒性的新型对策药物。