三种云南唐松草属植物中新颖生物碱的发现及其抗轮状病毒活性研究

The plants of Thalictrum genus have obvious medicinal values. More than 30 species of this genus had widely been used as folk medicine in China by 15 ethnic groups. The traditional drug use and published literature revealed that rich antiviral compounds might by existed in the plants of this genus. In our preliminary work, 43 samples of Thalictrum plants were collected in Yunnan Province based on the carefully survey of the folk using and resource distribution of the plants of this genus. With the guide of anti-rotavirus activity assay, three samples with obvious antiviral activity, Thalictrum glandulosissimum, Thalictrum finetii, and Thalictrum scabrifolium were selected out for further research. In addition, 13 new alkaloids (4 of them with notable anti-rotavirus activity) were isolated from Thalictrum glandulosissimum. The HPLC analysis showed that there is great possibility to find the novel bioactive alkaloids from the other two samples. In this project, the bioactive alkaloids in the active fractions of these three selected samples will be carefully studied by modern separation technology. As a result, more than thirty new alkaloids (incloding 3-5 compounds with new skeleton) were isolated from these samples. The antiviral activities of the isolated compounds will be screened, and 3-5 compounds with notable anti-rotavirus activity will be selected out. These researches will provide the scientific basis for the utilization of Yunnan folk medicine, and the development of new medicines with self-contained Intellectual Property for our country. Furthermore, the antiviral mechanism of some compounds with high activities will be studied, and the mechanisms of antiviral activities, the relationships of alkaloids structures andantiviral activities will be discussed in this project.

唐松草属植物具有广泛药用价值，我国有30多种该属植物在15个民族药中使用，民间用药和文献调研表明该属植物中有可能存在抗病毒活性成分。我们在对资源分布和民间用药充分调研的基础上，采集了云南43种唐松草属植物，以抗轮状病毒活性为导向，精选出金丝马尾连、滇川唐松草和鹤庆唐松草3个抗病毒活性显著的样品。通过对金丝马尾连的初步研究，发现了13个新颖的新生物碱，4个抗轮状病毒活性显著。HPLC分析结果表明另外两个样品中存在新颖活性生物碱的可能性也很大。项目将对三个样品活性部位中的生物碱进行深入研究，并对化合物进行活性筛选。预期从样品分离到30个以上的新颖生物碱，力争发现3-5个新骨架化合物或筛选出3-5个抗病毒活性突出的化合物结构；为合理开发利用云南民族药资源和开发我国自主知识产权的新药提供科学依据。项目还将选择部分活性好的化合物进行抗病毒机理和构效关系研究，揭示化合物的抗病毒活性机理和构效关系。

本项目围绕金丝马尾连 (T. glandulosissimum)、滇川唐松草 (T. finetii) 和鹤庆唐松草 (T. scabrifolium) 三种植物的研究，发现新生物碱类化合物33个，化合物结构类型包括异喹啉类、喹啉酮类、呋喃喹啉类、色酮[3,2-c]吡啶-10-酮类、苯并[c]氮杂-1-酮、苯扎地平类等。其中色酮[3,2-c]吡啶-10-酮类、苯并[c]氮杂-1-酮、苯扎地平类生物碱均为唐松草属植物中首次分离到。通过对化合物的抗轮状病毒活性筛选，发现4个治疗指数超过对照的化合物结构，还有6个化合物治疗指数和对照接近。为抗轮状病毒新药研发提供了新的药源分子。. 除上述三中植物外，本项目中还对星毛唐松草 (T. cirrhosum)、高原唐松草 (T. cultratum) 、丽江唐松草 (T. wangii)、小果唐松草 (T. microgynum) 四种唐松草属植物中的生物碱类化合物进行了扩展性研究，发现新生物碱类化合物11个，完成了化合物的抗轮状病毒活性筛选，发现治疗指数接近对照的化合物结构2个。发展和丰富类天然药物化学的内容。. 此外，项目组还通过扩展新研究，对决明属植物神黄豆 (C. agnes) 枝叶中分离的色酮类化合物、唇形科糙苏属植物假秦艽 (P. betonicoides) 中分离到的苯并异呋喃酮类化合物、茄科烟草属植物黄花烟草 (N. rustica) 中分离到的香豆素类化合物进行了抗轮状病毒活性筛选，未发现活性显著(治疗指数超过对照)的化合物结构。除抗轮状病毒活性外，项目组还对唐松草属植物中分离到的化合物进行了抗烟草花叶病毒、抗口腔有害微生物、抗甲氧西林耐药金黄色葡萄球菌、抗黑胫病等活性筛选，发现了一些活性显著的化合物。扩展了唐松草属植物活性成分的用途，为唐松草属植物综合开发利用提供了更多的科学依据。. 上述研究成果发表SCI论文12篇，已投稿5篇(其中3篇已通过审稿待发表)，申请发明专利7件，其中1件发明专利已获授权，完成了计划任务书规定的研究内容，并达到了预期目标。