PROTEIN DELIVERY INTO THE CENTRAL NERVOUS SYSTEM

治疗性多肽和蛋白质在血脑中的传递 进入中枢神经系统的屏障(BBB)已被证明是一种主要的 治疗神经系统疾病的障碍。选择性神经元丢失 在ALS、阿尔茨海默氏症、帕金森氏症、脑缺血和其他疾病中观察到 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有保护作用。 病变，支持了对多种疾病的治疗作用 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-1PNT-1等)。同样，超氧化物歧化酶， 脑源性神经营养因子和成纤维细胞生长因子被认为在预防海马区 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、扩散限制 脑室表面到脑实质，在那里这些因素 需要影响离散的退化神经元群体，并且迅速 证监会的批准强调了开发新技术的相当大的必要性 以非侵入性方式向神经系统输送药物以治疗脑血管疾病 人类的神经系统疾病。我们已经开发了方法论来 用来定量血脑屏障对多肽和蛋白质的通透性 对所占残余等离子体体积的适当修正 用第二种放射性示踪剂检测大脑毛细血管床中的蛋白质 同样的蛋白质。这项技术使我们能够开发出 促进治疗药物靶向神经系统递送的策略 肠外给药后的蛋白质。这些策略包括 具有高通透性的蛋白质的鉴定 提供治疗性化合物的载体。第二种策略是 对治疗性蛋白质进行化学或生化修饰以 在保持生物活性的同时增加渗透性。至关重要 对神经系统疾病的治疗不仅是增强的 治疗用药后血脑屏障通透性的变化 蛋白质，但也：1)必须保存蛋白质的生物活性 在修饰或偶联到载体之后，2)蛋白质必须 传递到受影响的神经元或神经胶质细胞的离散群体中 神经系统，3)蛋白质必须在分娩后保持其生物活性， 4)最重要的是，它必须能够在 这群细胞。在这项拨款建议中，我们计划测试 我们提高神经生长因子渗透性的策略的有效性 静脉给药后血脑屏障与天然蛋白的比较。这个 然后将评估产生最高渗透率的策略 三种动物模型来测试提供的改良的有效性 神经生长因子在离散脑内胆碱能神经元生物反应中的作用 肠外给药后的区域与天然NGF的比较。这些 研究结果将对人类神经系统疾病的治疗产生直接影响。 疾病。

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.