天然双吲哚生物碱色素多靶点抗阿尔兹海默病的研究

In our early research, we found that some red, black and blue pigments in several natural medicines are novel bis-indole alkaloids. Later, a series bioassay including anti Aβ and tau protein aggregation, anti-inflammatory, inhibiting acetylcholinesterase, inhibition of kinase, and oxidative stress, etc., revealed that these alkaloidal pigments are promising multi-targeted agents for Alzheimer’s disease. This project will set up the characteristic chromatographic-spectrum detection method using HPLC-DAD-MS/MS, fluorescence and UV, NMR technology, and quickly find their analogues from 12 kinds of related herb extracts. After isolation and identification of a variety of natural bis-indole alkaloid pigments, organic synthesis and biological transformation will be applied to built a diversified structure library. Subsequently, their anti-AD activities will be evaluated on molecular level, cellular level and animal level Alzheimer's disease models by using state of the art molecular biology techniques(ELISA/RTPCR/Western blot, etc.). The key pathways and molecular mechanisms and the ability as molecular probe of these alkaloids will be revealed. The 3D-QSAR model will be established to reveal their structure-activity relationship, and lead compounds for Alzheimer's disease will be found. Finally, virtual screening and docking will be carried out to investigate the interaction between these structures and key targets. The project will lay a solid foundation for discovery of novel drug leads and action mechanism for Alzheimer's disease.

前期发现多种天然药物中的红、黑、蓝色素为结构新颖的双吲哚生物碱，证实了他们通过抗Aβ和tau蛋白聚集、抗炎、抑制乙酰胆碱酯酶、抑制激酶、抗氧化应激等多重作用抗阿尔兹海默病。本课题拟利用HPLC-DAD-MS/MS、荧光和紫外、NMR等技术建立特征色谱光谱检测方法，从12种相关植物中快速发现类似物，分离鉴定多种天然双吲哚生物碱色素成分，并利用有机合成、生物发酵技术修饰转化构建多样结构库。运用分子药理学流行技术（ELISA/RTPCR/Western blot等），通过分子水平、细胞水平、动物水平的阿尔兹海默病相关模型，对此类物质进行药效药理研究，揭示其关键通路和分子水平作用机制，并探索此类色素的分子探针作用。建立3D-QSAR模型揭示其构效关系，发现先导化合物。阐明自然界存在的双吲哚生物碱色素抗AD的药效团、关键结构以及其分子机制和通路的关系。为发现活性更高/毒性更低的自主创新药物打下基础。

阿尔兹海默症的创新药物研发是当前的热点领域之一，由于其发病影响因素复杂，近年来不断有新的重要发现，需要紧跟研究前沿。本课题对马蓝、木蓝、大青、海州常山、赪桐、浙江大青等多种青黛、大青属植物中的化学成分进行了研究；对升麻属植物进行了双吲哚生物碱化学成分研究；并进一步对华泽兰、乌檀木、吴茱萸、远志正在进行生物碱研究，以期发现具有神经保护、抗AD疗效的先导化合物。目前共分离得到150种左右的化合物，但生物碱类较少。采用谷氨酸盐、丙烯醛诱导的海马神经元HT22细胞系损伤模型、MPP+诱导的SH-SY5Y神经细胞损伤模型等筛选评价了多种化合物的神经保护活性，并对部分化合物的机制进行了探讨。本项目将在当前研究基础上，继续双吲哚生物碱的发现和抗AD相关活性评价，并探索以调控溶酶体、胶质淋巴系统为方向的筛选评价方法，以期为发现具有新颖活性的抗AD药物先导物打下基础。