PROTEIN DELIVERY INTO THE CENTRAL NERVOUS SYSTEM

蛋白质输送到中枢神经系统

• 批准号: 2431303
• 负责人: JOSEPH F PODUSLO
• 金额: $ 28.96万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-06-01 至 1999-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2431303
• 关键词: PC12 cells; blood brain barrier; central nervous system; cerebral ischemia /hypoxia; choline acetyltransferase; enzyme activity; fibroblast growth factor; gene induction /repression; glycation; growth factor receptors; immunocytochemistry; laboratory mouse; laboratory rat; membrane permeability; messenger RNA; neuropharmacology; neurotrophic factors; nitric oxide synthase; protein tyrosine kinase; putrescine; spermidine; spermine; ubiquitin

Delivery of therapeutic peptides and proteins across the blood-brain barrier (BBB) into the central nervous system has proven to be a major obstacle in treating neurological diseases. The selective neuronal loss observed in ALS, Alzheimer's, Parkinson's, cerebral ischemia, and other neurodegenerative diseases, coupled with the growing body of evidence that neurotrophic factors have a protective effect against various degenerative lesions, has supported a therapeutic role for a large variety of neurotrophic factors and their derivatives in treating these diseases (e.g., NGF, BDNF, NT-3,4/5,CNTF, GDNF, IGF-1 PNT-1, etc.). Similarly, SOD, BDNF, and FGF have been suggested to have a role in preventing hippocampal neuronal damage following cerebral ischemia. The risk of infection, catheter clotting, neurosurgical costs, diffusional limitations beyond the ventricular surface to the parenchyma of the brain where these factors are needed to affect discrete populations of degenerating neurons, and rapid clearance by the CSF emphasize the considerable need to develop novel forms of non-invasive drug delivery to the nervous system for treatment of neurological diseases in humans. We have developed methodologies to quantify the permeability of the BBB to peptides and proteins with appropriate correction for the residual plasma volume occupied by the protein in the capillary bed of brain with a second radioactive tracer of the same protein. This technology has allowed our development of strategies to facilitate targeted nervous system delivery of therapeutic proteins after parenteral a ministration. These strategies include the identification of proteins with high permeabilities which could be used as carriers for the delivery of therapeutic compounds. A second strategy is the chemical or biochemical modification of therapeutic proteins to increase permeability while still preserving their bioactivity. Crucial to the treatment of neurological disease is not only the enhanced permeability at the BBB after parenteral administration of the therapeutic protein but also: 1) the bioactivity of the protein must be preserved after modification or coupling to a carrier, 2) the protein must be delivered to a discrete population of affected neurons or glia within the nervous system, 3) the protein must retain its bioactivity after delivery, and 4) most importantly, it must be capable of soliciting a bioresponse in this population of cells. In this grant proposal, we plan to test the efficacy of our strategies for increasing the permeability of NGF at the BBB compared to the native protein after parenteral administration. The strategy that produces the highest permeability will then be evaluated in three animal models to test the effectiveness of the delivered modified NGF in soliciting a bioresponse in cholinergic neurons in discrete brain regions after parenteral administration compared to the native NGF. These results will have direct implications for treating human neurological disease.

通过血脑输送治疗性肽和蛋白质 屏障（BBB）进入中枢神经系统已被证明是一个主要的 治疗神经系统疾病的障碍。选择性神经元丢失 在 ALS、阿尔茨海默病、帕金森病、脑缺血等疾病中观察到 神经退行性疾病，加上越来越多的证据表明 神经营养因子对各种退行性疾病具有保护作用 病变，已支持多种治疗作用 神经营养因子及其衍生物在治疗这些疾病中的作用 （例如，NGF、BDNF、NT-3,4/5、CNTF、GDNF、IGF-1 PNT-1 等）。同样，SOD， BDNF 和 FGF 被认为具有预防海马损伤的作用 脑缺血后的神经元损伤。感染的风险， 导管凝血、神经外科费用、超出治疗范围的扩散限制 这些因素所在的脑室表面到大脑实质 需要影响退化神经元的离散群体，并且快速 CSF 的批准强调了开发新型药物的巨大必要性 以非侵入性方式将药物输送至神经系统以治疗以下疾病 人类的神经系统疾病。我们开发了方法 量化 BBB 对肽和蛋白质的渗透性 适当校正所占用的残余血浆体积 大脑毛细血管床中的蛋白质与第二种放射性示踪剂 相同的蛋白质。这项技术使我们能够开发 促进有针对性的神经系统递送治疗的策略 胃肠外给药后的蛋白质。 这些策略包括 鉴定具有高渗透性的蛋白质，可用作 用于递送治疗化合物的载体。第二个策略是 治疗性蛋白质的化学或生化修饰 增加渗透性，同时仍保留其生物活性。 至关重要的 神经系统疾病的治疗不仅仅在于增强 胃肠外给予治疗剂后血脑屏障的通透性 蛋白质还：1) 必须保留蛋白质的生物活性 修饰或偶联至载体后，2) 蛋白质必须是 传递到受影响的神经元或神经胶质细胞的离散群体 神经系统，3) 蛋白质在递送后必须保持其生物活性， 4）最重要的是，它必须能够引起生物反应 这个细胞群。在此拨款提案中，我们计划测试 我们提高 NGF 渗透性的策略的有效性 胃肠外给药后 BBB 与天然蛋白的比较。这 然后将评估产生最高渗透率的策略 三种动物模型来测试所提供的修饰的有效性 NGF 在离散大脑胆碱能神经元中引发生物反应 与天然 NGF 相比，胃肠外给药后的区域。这些 结果将对治疗人类神经系统疾病产生直接影响 疾病。