基于AMPK调控通路和谱效整合技术定向捕获江西地产凤尾蕨属植物中抗糖尿病活性的蕨素类成分及构效关系研究

In recent years, the incidences of diabetes have been largely increasing. It is a worldwide and national threat. However, there hasn’t being a good drug for the disease in clinic by now. Therefore, the patients with diabetes have been crying for new drugs, which have good effect and low side effect. Some literatures showed that pterosins can significantly reverse the increased serum insulin and insulin resistance (IR) in IR mice and in db/db mice, and enhanced glucose uptake and Glut-4 mRNA expression in cultured IR cells. It’s interesting that our studies indicated a novel pterosin derivative, Bimutipterosins B, showed better anti-diabetic activity than the normal pterosin, pterosin A, which was recognized anti-diabetic pro-drug. So we will use the spectrum-effect integrated technology (HPLC-DAD-MSn-TD), which were established by our previous research, for screening the peterosins with antidiabetic effects from four plants of the genera Pter located in Jiangxi province, especial the nevol peterosin derivatives,. Furthermore, basing on the result of investigation for antidiabetic activity in vitro (AMPKα2β1γ1) and in vivo (IR mice and db/db mice), choosing peterosin derivatives with different structure glass as subjects, to study structure activity relationship, it can be completely investigated on structure activity relationship of pterosin derivatives. This item can supply some andiatetic activity constituents for studying and developing a new drug of disease of diabetes, and it can give foundation of the development of new antidiabetic drug of independent intellectual property rights.

近年来糖尿病发病率大幅度上升，目前治疗糖尿病的药物存在继发性失效、毒副作用明显及药价昂贵等诸多问题，不能很好地满足临床需求，因此临床上对新药物需求迫切。近期文献报道蕨素成分能激动AMPK而显示很好的抗糖尿病药物开发前景；本课题组前期研究发现，新型蕨素比常规蕨素更显著增加IR培养细胞的糖脂蛋白及Glut-4表达，提示新型蕨素抗糖尿病作用更优。据此，本项目拟以AMPKα2β1γ1为体外筛选靶标，采用谱效整合技术，结合正糖钳体内药效模型，研究江西地产凤尾蕨属植物中抗糖尿病活性的蕨素（重点新型蕨素类）成分；并在保持母核结构基本完整条件下，以支链酰基化入手逐渐过渡到A环取代基、B环共轭羰基研究顺序，再以C环立体构型为研究对象，全面考察各结构片段及构型对活性的贡献，探讨蕨素抗糖尿病活性的构效关系。通过本项目研究为抗糖尿病新药提供新先导化合物，为蕨素类成分开发新型抗糖尿病药提供理论基础。

目前糖尿病呈高发病率，而治疗其药物存在继发性失效和毒副作用明显等诸多问题，临床上对抗糖尿病新药需求迫切。文献及前期研究显示，凤尾蕨属植物中倍半萜成分（蕨素）具有较强的抗糖尿病作用，具潜在的抗糖尿病新药开发前景。本项目采用谱效整合技术，结合体内外筛选模型，对四种江西地产凤尾蕨属植物中抗糖尿病活性的蕨素成分开展了研究；并探讨了蕨素抗糖尿病构-效关系。通过上述研究，得出以下结果：1）从凤尾蕨属植物中共分离鉴定出100个蕨素化合物（包括井栏边草28个、欧洲凤尾蕨36个、蜈蚣草20个和江西凤尾蕨中16个），其中3个新蕨素二倍体和13个新蕨素；2）分离得到的蕨素类化合物体内外均显示不同程度的抗糖尿病作用；3）分析和归纳了蕨素天然产物的结构及其核磁共振谱特征；3）蕨素抗糖尿病活性与其结构存在以下关联性：不同取代基对目标化合物抗糖尿病活性影响较大，母核上羰基和C-3羟基为其抗糖尿病活性必要基团；C-3位羟基α构型较β活性强，糖苷化后活性增强；A环甲基被羟基取代、C-14位糖苷化及C-14/12和C-14/15形成六元环均使活性下降、蕨素二倍体C环甲基对位较同侧取代活性强。4）蕨素C 3-O-β-D-吡喃葡萄糖苷显示强的抗糖尿病活性，给药30小时体内降糖作用与二甲双胍相当，而48小时后，降糖作用比二甲双胍更优。.通过本项目的实施，培养了4名硕士研究生，3名本科毕业生，4名科研实践班学生；申请专利2项；目前已刊出学术论文5篇，1篇录用，其中SCI收录2篇，中文核心期刊收录4篇，3篇文章正处数据处理、撰写和投稿阶段。上述成果实现了本课题预定目标，分离鉴定出江西地产凤尾蕨属植物中抗糖尿病活性的蕨素成分，丰富了天然蕨素类化合物的类型，为抗糖尿病新药提供了新的先导化合物。初步阐明了蕨素类化合物抗糖尿病活性的构-效关系，为蕨素类化合物开发为新型抗糖尿病药提供了理论基础。