WGA as a Novel Vehicle for Intranasal Delivery of an Anti-A-beta Antibody in AD

WGA 作为 AD 中抗 A-β 抗体鼻内递送的新型载体

• 批准号: 8432312
• 负责人: NEELIMA CHAUHAN
• 金额: $ 7.8万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2014-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8432312
• 关键词: Active Immunization; Affect; Afferent Neurons; Alzheimer disease prevention; Alzheimer' s Disease; American; Amyloid; Anatomy; Antibodies; Area; Asses; Attention; Attenuated; Axon; Axonal Transport; Basal Cell; Binding; Biochemical; Bipolar Neuron; Blood; Blood - brain barrier anatomy; Blood Vessels; Brain; Bypass; Carrier Proteins; Cephalic; Cerebrum; Characteristics; Chitosan; Cilia; Cleaved cell; Clinical Trials; Cognitive; Cognitive deficits; Development; Diffusion; Disease; Edema; Endocytosis; Epithelial; Excision; Gel; Glucosamine; Goals; Grant; Human; Immunization; Immunotherapeutic agent; Immunotherapy; Intracellular Transport; Intranasal Administration; Lead; Lectin; Liposomes; Liquid substance; Mediating; Methodology; Microspheres; Minor; Modeling; Mono-S; Monoclonal Antibodies; Mus; Nasal cavity; Neuraxis; Neurodegenerative Disorders; Neuroglia; Nose; Olfactory Epithelium; Olfactory tract; Passive Immunization; Pathway interactions; Penetration; Peptide Hydrolases; Pharmaceutical Preparations; Phase; Physiological; Prevention; Process; Resources; Route; Safety; Sialic Acids; Side; Staging; Structure of mucous membrane of nose; Subarachnoid Space; Supporting Cell; Surface; System; Testing; Therapeutic; Transgenic Mice; Translating; Wheat Germ Agglutinins; Wild Type Mouse; absorption; amyloid peptide; bile salts; cognitive change; cribriform plate; dimer; drug candidate; drug discovery; efficacy testing; extracellular; improved; intravenous injection; mouse model; novel; olfactory bulb; preclinical study; preference; prevent; receptor mediated endocytosis; response; surfactant; trafficking; uptake

This application is in response to PAS-10-151 {Grants for Alzheimer's Disease Drug Discovery (R21)}. Novel vehicles are needed to effectively target direct delivery of immunotherapeutics to the brain in order to maximize the efficacy of passive immunization for treating, delaying or preventing Alzheimer's disease (AD), a disease that currently afflicts >5 million Americans. Conjugation of the lectin, Wheat Germ Agglutinin (WGA), has been shown to enhance intranasal uptake of candidate drugs and promote direct delivery to brain via (i) Adsorptive or receptor-mediated endocytosis into the olfactory sensory neurons (OSN) followed by intracellular transport to olfactory bulb; (ii) Non-specific fluid phase endocytosis into the OSNs followed by intracellular transport to olfactory bulb; and/or (iii) Extracellular diffusion along the inter-olfactory epithelial clefts directly to the olfactory bulb and/or CSF. Passive immunization by intravenous injection of Ass monoclonal antibodies (mAb) has shown promising results such as lowering of cerebral Ass and stabilization or prevention of cognitive deficits in preclinical studies in transgenic mouse models of AD and in early human clinical trials. However, the level of antibody penetration in the brain is limited by the blood-brain-barrier (BBB). In addition, in mice and humans with vascular amyloid, high levels of anti-Ass antibodies in blood has led to microhemorrhage and vasogenic edema. This project aims to overcome such limitations by maximizing the efficacy of the intranasal route of anti-Ass antibody delivery using a WGA carrier protein. This project will test the efficacy of WGA as a novel vehicle that will not only enhance delivery of anti-Ass mAb, 3A1, that binds to mono/dimers and fibrillar Ass (To be provided by Dr. Lemere-Consultant) by virtue of its endocytic uptake preference, but will also facilitate passive diffusion of 3A1 due to its nasal mucosa-like biochemical composition, maximizing immunotherapeutic efficacy of intranasal passive immunization, to be tested in Alzheimer's 5XFAD model. Hypothesis: Intranasal delivery of WGA-conjugated 3A1 mAb combined with unlabeled 3A1 antibody will maximize the immunotherapeutic potential of intranasal passive immunization ameliorating AD-like patho-cognitive changes in 5XFAD transgenic mice. Specific Aims: [1] Trafficking of WGA-3A1 antibody after intranasal delivery to the brain in wild type mice and binding of 3A1-WGA to Ass plaques in the brains of 5XFAD mice and human AD. [2] Determining if intranasal passive immunization with WGA-conjugated 3A1 mAb combined with unlabeled 3A1 antibody will prevent AD-like patho-cognitive changes in young pre-plaque 5XFAD mice. [3] Determining if intranasal passive immunization with WGA-conjugated 3A1 mAB combined with unlabeled 3A1 antibody will attenuate AD-like patho-cognitive changes in old late-stage plaque-bearing 5XFAD mice. All resources, including transgenic mice, and required expertise are ready to begin this study. Significance: This project will evaluate a novel delivery system to target immunotherapeutics directly to the brain, and thus advance one of the most promising therapies for delaying the progression or prevention of AD.

本申请是对PAS-10-151 {阿尔茨海默病药物发现（R21）赠款}的回应。 需要新的载体来有效地将免疫治疗剂直接靶向递送至大脑，以便 最大限度地提高被动免疫治疗、延缓或预防阿尔茨海默病（AD）的功效， 这种疾病目前困扰着超过500万美国人。凝集素，麦胚凝集素（WGA）， 已经显示增强候选药物的鼻内摄取并促进通过（i）直接递送至脑， 通过吸附或受体介导的内吞作用进入嗅感觉神经元（OSN），然后在细胞内 （ii）非特异性液相内吞进入OSN，随后细胞内吞进入OSN。 运输至嗅球;和/或（iii）沿着嗅间上皮裂隙直接细胞外扩散至 嗅球和/或脑脊液Ass单克隆抗体静脉注射被动免疫 (mAb)已经显示出有希望的结果，如降低大脑的Ass和稳定或预防认知功能障碍。 在AD转基因小鼠模型和早期人类临床试验的临床前研究中存在缺陷。但 抗体在脑中的渗透水平受到血脑屏障（BBB）的限制。此外，在小鼠和 患有血管淀粉样蛋白的人，血液中高水平的抗Ass抗体导致微出血， 血管源性水肿该项目旨在通过最大限度地提高鼻内给药的疗效来克服这些局限性。 使用WGA载体蛋白的抗Ass抗体递送途径。该项目将测试WGA作为一种 一种新型载体，不仅能增强抗Ass mAb 3A 1的递送，3A 1结合单聚体/二聚体和纤维状Ass (To由Lemere-Consultant博士提供），由于其内吞摄取偏好，但也将促进 由于其鼻粘膜样生化成分，3A 1的被动扩散，最大限度地提高免疫原性 鼻内被动免疫的功效，将在阿尔茨海默氏症5XFAD模型中进行测试。假设：鼻内 与未标记的3A 1抗体组合的WGA缀合的3A 1 mAb的递送将最大化 鼻内被动免疫改善AD样病理性认知改变的免疫调节潜力 在5XFAD转基因小鼠中。具体目的：[1]在鼻内递送至受试者后WGA-3A 1抗体的运输。 在野生型小鼠的脑中，3A 1-WGA与5XFAD小鼠和人AD的脑中的Ass斑块的结合。[二]《中国日报》 确定是否用WGA缀合的3A 1 mAb与未标记的3A 1组合进行鼻内被动免疫 抗体将预防幼年斑块前5XFAD小鼠中的AD样病理性认知变化。[3]确定是否 用WGA缀合的3A 1 mAB与未标记的3A 1抗体组合的鼻内被动免疫将 减弱老年晚期斑块携带5XFAD小鼠中AD样病理认知变化。所有资源， 包括转基因小鼠和所需的专业知识准备好开始这项研究。意义：该项目将 评估一种新的递送系统，将免疫治疗剂直接靶向大脑，从而推进 延缓或预防AD的最有希望的疗法。