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; 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 有机磷酸盐（OP）化合物包括农药和化学战神经剂。他们很高 有毒并可能产生胆碱能危机，迅速发展为癫痫持续状态（SE），甚至死亡 没有紧急护理。用阿托品，pralidoxime和咪达唑仑治疗的标准护理（SOC）治疗 OP中毒后，大大改善了生存。然而，许多OP-SE的幸存者表现出脑损伤， 认知障碍和自发癫痫发作（SRS）。另外，急性和持久 据报道，在OP之后，已经报道了神经炎症和促炎细胞因子的表达增加 se。这些持续性神经炎性变化被认为是神经变性的基础，网络超级 - 兴奋性和适应不良的可塑性，导致认知功能障碍和SRS。因此，缓解神经 炎症是减轻神经元损伤和行为病因的主要目标。 Verapamil（VPM）是用于高血压和心绞痛治疗的水溶性钙通道阻滞剂。 最近的发现还证明了VPM的有效神经保护作用和抗炎作用 各种中枢神经系统受伤。我们的初步结果表明，肌内VPM（10 mg/kg，I.M.）是安全的，并且 当多个大脑区域中产生明显的神经保护和神经炎症的降低 DFP SE终止后进行管理。它也与促炎和降低有关 抗炎细胞因子的上调。最后，由于VPM提高了这种效果，因此具有功能性结果 DFP后八周的焦虑和认知障碍。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 领域。研究将采用DFP-SE大鼠模型来执行具体目的：在AIM 1中，重复的安全性 我是。将评估注射剂，并评估药代动力学参数和稳定性 VPM公式。在AIM 2中，VPM治疗对减少神经元死亡和神经炎症的影响 在DFP之后，将评估使用氟-jade C以及神经胶质原纤维酸性蛋白和电离 钙结合衔接子分子-1免疫组织化学染色。我们还将评估效果 VPM治疗促和抗炎细胞因子表达。在AIM 3中，VPM治疗将通过 研究各种VPM剂量和治疗持续时间对神经降解的影响 神经炎症。在AIM 4中，对长期焦虑的优化VPM治疗的功能结果， 认知障碍和SRS将使用啮齿动物行为分析和脑电图技术进行测试。 我们还将在研究结束时起草初步的目标产品概况（TPP）。这些研究会 进一步了解神经炎症机制在介导的疾病和 优化基于VPM的抗炎疗法作为一种减轻OP毒性的新型对策药物。