山蜡梅具抗流感病毒活性的新颖生物碱成分研究

Chimonanthus nitens Oliv. is the unique plant (family Calycanthaceae) in China, it is mainly distributed in the northeast of Jiangxi and the southeast of Anhui province, for example, Damaoshan located in Dexing, Yushan located in Wuyuan, Qinyunshan and Huizhoushan located in Anhui, and so on. Chimonanthus nitens was used in traditional Chinese medicine (TCM) to treat cold, bacterial infection, inflammation for long time in folk, and the aerial parts of this plant and its leaves were sold at all the streets in Jiangxi Wanyun when it is the dragon boat festival. Some pharmaceutical companies have marketed some produces, such as Shanlamei Keli, Shanlamei Pian and Shanlamei Diwan. To the best of our knowledge, there are some literatures about research on constituents of volatile oil from Chimonanthus nitens, but a little research on anti-virus activity have been reported. We did some research on the plant for many years, and we found that the alkaloids isolated from the plant were principles of anti-virus activity. As we all know, alkaloids include most of novel natural products, and alkaloids isolated from plants usually have obvious physiological activities, so it is the hot subject to some natural product researchers. So based on our fundamental works, we will focus on studying on the alkaloids with anti-virus activity from Chimonanthus nitens in this proposal, and hunting the lead compounds of anti-virus activity. The results will provide scientific and theoretical basis for further developing and using Chimonanthus nitens of Jiangxi advantage herb.

山蜡梅为我国特有的蜡梅科蜡梅属植物，主要分布于江西德兴大茅山区、婺源怀玉山区及安徽徽州山区齐云山一带。江西婺源民间用山蜡梅茶（俗称茅山茶）防治感冒、抗菌消炎已有几百年的历史。每年的端午节前后，街头巷尾和菜市场等处，时时可见成捆的山蜡梅枝叶出售，生意兴隆。江西相关制药企业已经先后生产销售山蜡梅叶颗粒、山蜡梅叶片等感冒用药，具有辛凉解表，清热解毒等功效。至今为止山蜡梅挥发油成分的研究报导较多，未见其抗病毒等活性成分的文献。我们对山蜡梅植物进行了多年研究，发现其总生物碱成分具有较好的抗流感病毒作用。 生物碱类蕴藏着大量结构新颖的天然有机化合物，植物中存在的生物碱大多具有明显的生理活性，是天然药物学化学家研究的热门领域。本项目旨在扎实的前期工作基础之上，重点研究山蜡梅中具有抗流感病毒活性的生物碱类成分，筛选出具有抗流感活性的药物先导化合物，为山蜡梅这一江西优势药材的广泛应用提供科学根据。

山蜡梅为我国特有的蜡梅科蜡梅属植物，主要分布于江西德兴大茅山区、婺源怀玉山区及安徽徽州山区齐云山一带。江西婺源民间用山蜡梅茶（俗称茅山茶）防治感冒、抗菌消炎已有几百年的历史。本课题根据民间用药习惯，建立了在现代抗H1N1 流感病毒药理活性指引下，系统研究了该植物的化学成分。建立了生物碱的含量测定方法，富集到得到总碱含量大于50%的抗流感病毒的总生物碱有效部位。优化了总碱的提取纯化工艺及挥发油的提取方法。率先探索其抗H1N1 流感病毒的药效物质基础，阐明了植物中的生物碱和挥发油成分是抗H1N1 流感病毒的主要活性成分，其中以挥发油的效果更佳。比较了4、5、6、7四个月份药材挥发油的质量，证实了山蜡梅药材的最佳采收季节为每年的6月份。为江西民间清明节前后采药的习惯提供了科学根据。. 建立了抗流感病毒 H1N1 药物筛选模型，利用 MDCK 细胞对甲型流感病毒[A/PR/8/34H1N1]进行传代培养的基础上，建立细胞病变模型，并利用此模型指导山蜡梅化合物的分离精制，分离得到77个化学成分，确定了其中55个化合物的结构，有6个是结构新颖的化合物,其余化合物的结构待定。首次从该蜡梅属植物中分到的化合物12个，首次从该山蜡梅植物中分到的化合物8个。为山蜡梅植物的生物分类学研究奠定了基础。. 筛选到10个生物碱成分在所检测浓度范围内对H1N1 PR8在MDCK细胞中的复制具有一定的抑制效果，其中6个可作为药物开发的先导化合物继续深入研究。为山蜡梅植物作为抗流感民间用药提供了科学依据。