基于CHaC功能化修饰的姜黄素脑靶向转运体胶束的构建及其抗阿尔茨海默病研究

Nowadays, the prevalence rate of AD has significantly rose which brings a great burden to the patient and the society. The blood–brain barrier behaves as the main obstacle to AD and contributes to poor therapeutic of lot of drugs because of low concentration in brain. Micelles is interesting due to its basic functionality and the advantages that ensue. Choline transporter-like protein 1 (CTL1), a kind of transporter expressed in brain, has specific affinity with choline and its structural congeners (CHaC). It's worth noting that CTL1 has not been used to enhance brain uptake of micelles by specific interaction with choline and its structural congeners (CHaC) as a ligand in surface of brain microvascular endothelial cell. Moreover, it is not clear that the transport mechanisms based on specific interaction with CTL1 in brain. In view of the advantages of micelles in drug delivery for Alzheimer's disease, glioma therapy, and so on, it is necessary to explore new specific affinity site in surface of brain microvascular endothelial cell to promote drug uptake in micelles. At present, we have successfully prepared CHaC- functionalized curcumin loaded micelles so called transmicelles. The brain distribution of curcumin was increased by 4.18-fold in mice. This project is designed to complete by following experiments: 1. synthesizing PEG-GCDS containing choline and its structural congeners（CHaC -PEG-GCDS）; 2. preparing and evaluating curcumin transmicelles containing different density of choline and its structural congeners; 3. conducting blood-brain barrier penetration and anti-Aβ experiment to study transport mechanism and AD therapy; 4. investigating the pharmacokinitics behaviors and in vivo anti-AD activity of transmicelles. This research is trying to clarify the transport mechanisms of CHaC- functionalized micelles targeting CTL1 in brain and provide a theoretical basis for the transporter as a target for drugs delivery and for the treatment of AD.

近年来，阿尔茨海默病（AD）患病率显著增长，血脑屏障给AD的治疗带来了极大的挑战。胶束在提高药物脑内传递方面优势明显。胆碱转运体类蛋白1（CTL1）是脑部转运体的一种，对胆碱及其同系物（CHaC）具有高度的选择性。以CHaC为配基，CTL1为靶点提高胶束的脑部转运还未见报道，转运机制尚不明确。鉴于胶束在AD等疾病治疗中递药优势明显，寻找新靶点并明确转运机制是十分必要的。目前我们已成功制备了CHaC修饰的姜黄素载药胶束，小鼠组织分布表明其能够将姜黄素脑内浓度提高4.18倍。本课题拟完成：1.合成不同的CHaC-PEG-GCDS；2.制备不同CHaC密度的姜黄素转运体胶束并进行评价；3.采用血脑屏障转运实验和抗Aβ作用解析跨膜转运和AD治疗机制；4.考察体内药动学和抗AD效果。通过研究，阐明CHaC功能化修饰的脑靶向胶束增加药物转运的机制，为转运体胶束的构建及AD治疗提供理论依据和靶向策略。

本课题紧紧围绕基于CHaC功能化修饰的姜黄素脑靶向转运体胶束的构建及其抗阿尔茨海默病研究开展工作。合成了聚乙二醇-二硬脂酸甘油酯，ChaC-聚乙二醇-二硬脂酸甘油酯，构建了基于胆碱及其同系物（CHaC）功能化修饰的姜黄素脑靶向转运体胶束所需的载体材料；利用合成的载体材料，制备了姜黄素自组装的空白胶束和转运体胶束，并完成了粒径测定，形态观察，包封率和释放度等体外表征；使用脑微血管内皮细胞细胞模型，通过密胆碱竞争性抑制实验、细胞摄取实验和细胞转运实验，揭示了进入细胞均属于能量依赖型的，其中空白胶束进入细胞是由部分网格蛋白还和部分巨胞饮途径介导的，转运体胶束是由部分网格蛋白、部分巨胞饮和部分非网格蛋白和非细胞膜穴样内陷介导途径的；采用高效液相色谱质谱联用法测定姜黄素大鼠血浆中的血药浓度并进行方法学验证，应用于姜黄素空白胶束和转运体胶束大鼠静脉注射的药动学和组织分布研究；建立大鼠阿尔茨海默病模型，研究空白胶束和转运体胶束的抗AD活性。通过研究，阐明CHaC功能化修饰的脑靶向胶束增加药物转运的机制，为转运体胶束的构建及AD治疗提供理论依据和靶向策略。截至到目前，本课题相关内容已获批宁夏医科大学总医院（科研平台）开放课题研究项目1项，现有专利1项和2篇SCI文章正在撰写。.