Targeting toll like receptor 4 (TLR4) and TLR2 to resolve EAE-associated paralysis, pain and cognitive deficits: efficacy of a clinically-relevant blood brain barrier permeable TLR4/TLR2 antagonist

靶向 Toll 样受体 4 (TLR4) 和 TLR2 以解决 EAE 相关的麻痹、疼痛和认知缺陷：临床相关血脑屏障可渗透性 TLR4/TLR2 拮抗剂的功效

• 批准号: 9153350
• 负责人: LINDA WATKINS
• 金额: $ 33.68万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-15 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9153350
• 关键词: Address; Anti-Inflammatory Agents; Anti-inflammatory; Behavioral; Binding; Blood - brain barrier anatomy; Brain; Cell Death; Cells; Cerebrospinal Fluid; Characteristics; Chronic; Clinical; Clinical Trials; Cognitive deficits; Demyelinations; Department of Defense; Development; Disease; Disease Progression; Dose; Drug abuse; Encephalitis; Experimental Autoimmune Encephalomyelitis; Female; Functional disorder; Funding; Goals; Grant; Height; Human; Immune; Immunohistochemistry; Immunologic Receptors; Impaired cognition; Impairment; Inflammation; Inflammatory; Investigation; Investigational Drugs; Investigational New Drug Application; Isomerism; Life; Literature; Lysophosphatidylcholines; Maps; Modeling; Motor; Multiple Sclerosis; Naltrexone; Opioid; Opioid Receptor; Pain; Pain management; Paralysed; Paresis; Pathology; Population; Process; Rattus; Recruitment Activity; Research; Rodent; Role; Series; Site; Social Interaction; Specificity; Spinal; Spinal Cord; Symptoms; System; TLR2 gene; TLR4 gene; Testing; Therapeutic; Tissues; Translations; Work; behavioral study; chronic demyelination; clinically relevant; cytokine; glial activation; improved; male; motor disorder; multiple sclerosis treatment; neuroinflammation; neuron loss; non-drug; novel; painful neuropathy; pre-clinical; receptor; remyelination; small molecule; social cognition; symptomatic improvement

PROJECT SUMMARY Multiple sclerosis (MS) is a life-long, debilitating disease in both males and females. Symptoms include loss of motor function, neuropathic pain, cognitive impairments, and impaired social interaction. MS is furthermore associated with elevations in circulating and central (spinal cord and brain) levels of pro- inflammatory cytokines and decreased levels of anti-inflammatory cytokines, suggesting a dysregulation of immune and glial processes resulting in chronic inflammation. This ongoing inflammation is important in demyelination, chronic glial activation, and neuronal death characteristic of the disease. Targeted suppression of spinal cord neuroinflammation using anti-inflammatory strategies dramatically improves symptoms of experimental autoimmune encephalomyelitis (EAE), a rat model of MS. Looking forward toward translation, what is needed is not our current approaches that are injected intrathecally, but rather a means to effectively treat EAE/MS via a clinically relevant, orally available, blood-brain barrier permeable small molecule that targets EAE/MS pathology driven by neuroinflammation. We have discovered, and extensively characterized, such a small molecule. This drug (the non-opioid (+)-isomer of naltrexone; (+)-naltrexone) is rapidly moving toward FDA application for Investigational New Drug status. This is a selective antagonist at toll-like receptor 4 (TLR4) and TLR2. As it fails to bind classical opioid receptors, (+)-naltrexone does not interfere with the efficacy of opioids for pain control or normal functioning of opioid receptor systems. Targeting TLR2 and TLR4 arises from an extensive literature demonstrating the importance of these receptors in the neuroinflammatory processes and EAE/MS symptoms to be studied here. A complimentary series of behavioral and immunohistochemistry studies are proposed in males and females which will explore the ability of (+)-naltrexone to suppress EAE-induced (a) paresis/paralysis, (b) neuropathic pain, (c) cognitive impairment, and (d) social interaction impairment, as well as (e) improve survival, when (+)-naltrexone is systemically administered across days, comparing dosing early vs. late in the EAE timecourse. The IHC studies will analyze brain and spinal cord tissues collected after early vs. late (+)- naltrexone treatment (mapping onto the behavioral studies) to define whether (+)-naltrexone suppresses glial activation, neuronal cell death, and demyelination, as well as stimulates remyelination, as predicted. These studies create two complimentary Aims. Aim 1 explores a novel means of positively intervening in EAE-induced motor dysfunction, neuropathic pain, deficits in social interaction and cognition, and loss of life by targeting TLR4/TLR2 by systemic administration of (+)-naltrexone. Aim 2 transitions to an initial exploration of potential mechanisms underlying pathophysiology, focusing on the impact of TLR4/TLR2 blockade on demyelination and remyelination, neuronal cell death, and neuroinflammation. These studies provide a thorough investigation of the role of TLR4/TLR2 in major MS-relevant symptoms of EAE.

项目摘要 多发性硬化症（MS）是一种终身的，使人衰弱的疾病，男性和女性都有。症状包括 运动功能丧失、神经性疼痛、认知障碍和社会交往障碍。MS是 此外，与循环和中枢（脊髓和脑）前体水平升高有关， 炎性细胞因子和抗炎细胞因子水平降低，表明 免疫和神经胶质过程导致慢性炎症。这种持续的炎症对于 脱髓鞘、慢性神经胶质活化和疾病特征性的神经元死亡。 使用抗炎策略靶向抑制脊髓神经炎症 改善实验性自身免疫性脑脊髓炎（EAE）的症状，这是一种MS大鼠模型。 对于翻译，需要的不是我们目前的鞘内注射方法，而是一种 通过临床相关、口服、血脑屏障可渗透的小分子药物有效治疗EAE/MS 靶向由神经炎症驱动的EAE/MS病理学的分子。 我们已经发现并广泛表征了这样一种小分子。这种药物（非阿片类药物） （+）-纳曲酮异构体;（+）-纳曲酮）正在迅速向FDA申请研究性新药 status.这是Toll样受体4（TLR 4）和TLR 2的选择性拮抗剂。因为它不能结合经典的阿片类药物 受体，（+）-纳洛酮不干扰阿片类药物的疼痛控制或正常功能的疗效， 阿片受体系统靶向TLR 2和TLR 4来自大量文献，其证明了 这些受体在神经炎症过程和EAE/MS症状中的重要性有待研究。 一个免费的一系列行为和免疫组织化学研究提出了在男性和 探索（+）-纳洛酮抑制EAE诱导的（a）轻瘫/麻痹，（B）的能力的雌性动物 神经性疼痛，（c）认知障碍，和（d）社会互动障碍，以及（e）改善 生存期，当（+）-纳洛酮在几天内全身给药时，比较早期与晚期给药 EAE时间进程。IHC研究将分析早期与晚期（+）- 纳洛酮治疗（映射到行为研究），以确定（+）-纳洛酮是否抑制神经胶质细胞增殖， 激活、神经元细胞死亡和脱髓鞘，以及刺激髓鞘再生，如预测的那样。 这些研究创造了两个互补的目标。目标1探索了一种积极干预的新方法 EAE诱导的运动功能障碍、神经性疼痛、社会互动和认知缺陷以及 通过全身施用（+）-纳洛酮靶向TLR 4/TLR 2。目标2过渡到初步探索 潜在的病理生理学机制，重点是TLR 4/TLR 2阻断对 脱髓鞘和髓鞘再生、神经元细胞死亡和神经炎症。这些研究提供了一个 深入研究TLR 4/TLR 2在EAE的主要MS相关症状中的作用。