【作者】 梁冬冬；

【导师】 刘珂；

【作者基本信息】 吉林大学 ， 生物化学与分子生物学， 2008， 硕士

【摘要】 从传统中药中寻找具有治疗潜力的先导化合物,再进行结构优化提高成药性,从而获取候选药物是我国自主知识产权创新药物研究的重要途径之一。豨莶草(Herba siegesbeckiae)是菊科(Compositae)豨莶属(Siegesbeckia)一年生草本植物,可以治疗风湿痹痛、筋骨无力、腰膝酸软、四肢麻痹、半身不遂、风疹湿疮。此外,尚有镇静、安神、明目作用。奇壬醇是豨莶草中的主要有效成分,具有祛风湿、抗炎、增强免疫等作用,为中药中治疗风湿性关节炎的活性化合物之一。我们以奇壬醇为母体,通过结构修饰,设计并合成了脂溶性衍生物并考察了其抗炎活性。本实验主要由三部分组成。第一部分:考察了不同产地的豨莶草中奇壬醇的含量,提取分离奇壬醇,为结构修饰提供原料。第二部分:对奇壬醇进行缩醛化、缩酮化、乙酰化、甲醚化等一系列的结构修饰,得到23个目标产物:二甲基-21-亚甲叉基奇壬醇、环己基-21-亚甲叉基奇壬醇、β-乙基-α-甲基-21-亚甲叉基奇壬醇、α-乙基-β-甲基-21-亚甲叉基奇壬醇、β-异丙基-α-甲基-21-亚甲叉基奇壬醇、α-异丙基-β-甲基-21-亚甲叉基奇壬醇、β-异丁基-α-甲基-21-亚甲叉基奇壬醇、α-异丁基-β-甲基-21-亚甲叉基奇壬醇、二苯基-21-亚甲叉基奇壬醇、β-(3,4-二甲氧基)苯甲基-21-亚甲叉基奇壬醇、α-(3,4-二甲氧基)苯甲基-21-亚甲叉基奇壬醇、β-对羟基苯甲基-21-亚甲叉基奇壬醇、α-对羟基苯甲基-21-亚甲叉基奇壬醇、β-(2-羟基)苯甲基-21-亚甲叉基奇壬醇、α-(2-羟基)苯甲基-21-亚甲叉基奇壬醇、β-(3-甲氧基-4-羟基)苯甲基-21-亚甲叉基奇壬醇、α-(3-甲氧基-4-羟基)苯甲基-21-亚甲叉基奇壬醇、2,15,16,19-四乙酰基奇壬醇、2,16,19-三乙酰基奇壬醇、16,19-二乙酰基奇壬醇、2,15,16-三甲氧基奇壬醇、15,16,19-三甲氧基奇壬醇、15,16二甲氧基奇壬醇。第三部分:因为一氧化氮(NO)是关节炎病发过程中一种重要的炎症介质,具有促进炎症和抗炎的双重特性,所以,我们将奇壬醇衍生物对LPS诱导的巨噬细胞NO抑制活性进行评价及以期寻找到新的具有更好药理活性的先导化合物。结果显示,奇壬醇衍生物的活性与化合物分子量无关,主要随被修饰羟基的个数及基团位置的变化而变化。随着被修饰羟基个数的增加,其衍生物活性越好;基团空间取向的不同也会导致活性有所差异。更多还原

【Abstract】 Base on the discovery of lead compounds which have therapeutic value from the traditional Chinese herb, then modify them to enhance its curative effect is an important way to gain candidate drugs which have China’s own intellectual property rights.Arthritis is a common disease, and rheumatoid arthritis is the most common and difficult to cure. The ratio of disability is quite high. Traditional drugs which are used to treatment rheumatoid arthritis are most of NSAIDs and COX-2 selective inhibitors. However, the serious gastrointestinal (GI) adverse reactions and the side effects of ardiovascular which are discovered limit their further application. In recent years, the advent of NO-NSAIDs to anti-inflammatory drugs brought significant progress. Such drugs can both inhibit COX to anti-inflammatory or analgesic and protect blood vessel by GI with NO-induced. So far, there are several NO-NSAIDs come into different stages of clinical research and the research of this area is becoming hotter.NO is a small molecule that found from endothelial vascular, it is synthesized by L-arg with NOS-induced. The structure of NO is simple but extremely unstable, easy to proliferation, easy to react and the biological half-life is only about 5s.The biosynthesis of NO in vivo is like this: by the catalytic of NOS the L-arg is oxidized and brings Citrulline, at the same time release NO. The research shows that there are relationships between acute or chronic inflammation and NO. The source of NO is not clear by now, may be from blood vessels, neutrophil or macrophage. The anti-inflammatory of NO is mainly in the following aspects: First, There is lot of NO in the host with inflammatory response and they can prevent the entrance of cells, viruses and parasites. Second, to inhibit inflammation, the NO maintain the integrity of BVE by adjusting the relationships between platelet - intramural vascular and inhibit the aggregation or depolymerization of platelet. Third, it can inhibit the proliferation of lymphocyte and at the same time stimulation many immune responses or immune pathological process including of rheumatoid arthritis.Siegesbeckiae orentalis(Compositae)is a herbaceous plant that can be used to treat rheumatism, inflammatory and other disease. Kirenol is a main active component of siegesbeckia orentalis, and it can be used to anti-rheumatism, anti-inflammatory and enhance immunity. It is a lead compound of anti-rheumatism which is used as a traditional Chinese herb.But the research of kirenol is most about its mechanism of anti-inflammatory, rarely about the modification of it.So, we modified it with the aim of gaining the compounds that has better liposolubility and better bioavailability than kirenol, and the inhibitory activity on nitric oxide (NO) production in LPS-activated macrophage were tested.First, we can see that the content of kirenol in different origin of Herba Siegesbeckiae is different from the articles before. Therefore, we test the content of kirenol by HPLC and selected the best one. By comparison, we selected 5kg Herba Siegesbeckiae from Longkou in Shandong province, and then isolated 4.208 kirenol. Then, we modified them. We design a series of acetal and ketal reaction on kirenol, because many drugs contain acetal or ketal groups which can significantly improve drug activity. By these, we got 23 compounds: Dimethyl-21-ethenetylene-kirenol, cyclohexyl-21- ethenetylene- kirenol,β-ethyl-α-methyl-21- ethenetylene- kirenol,α-ethyl-β-methyl-21- ethenetylene- kirenol,β-isopropyl-α-methyl-21- ethenetylene- kirenol,α-isopropyl-β-methyl-21- ethenetylene- kirenol,β-isobutyl-α-methyl-21- ethenetylene- kirenol,α-isobutyl-β-methyl-21- ethenetylene- kirenol, diphenyl-21- ethenetylene- kirenol,β- ( 3 , 4-dimethoxy ) benzyl-21- ethenetylene- kirenol,α-(3,4-dimethoxy)benzyl-21- ethenetylene- kirenol,β-p-hydroxy-benzyl -21- ethenetylene- kirenol,α- p-hydroxy -benzyl -21- ethenetylene- kirenol,β-(2-hydroxy)benzyl-21- ethenetylene- kirenol,α-(2-hydroxy)benzyl-21- ethenetylene- kirenol,β-(3-methoxy-4-hydroxy)benzyl -21- ethenetylene- kirenol,α-(3-methoxy-4-hydroxy)benzyl -21- ethenetylene- kirenol, 2,15,16,19- quadracetyl- kirenol、2,16,19- triacetyl- kirenol, 16 , 19- diacetyl- kirenol, 2,15,16-trimethoxy- kirenol, 15,16 ,19-trimethoxy- kirenol, 15,16-dimethoxy- kirenol, and 19 of them were first synthesized. There are several enantiomers. To establish the exact structure of them, two-dimensional NMR experiments including COSY, HSQC, and HMQC were carried out. Finally, their inhibitory activity on NO production in LPS-activated mouse macrophage was tested with the aim of gain lead compounds that more active than kirenol.The result shows that the activity of kirenol derivatives is nothing about molecular weight but the number of hydroxyl which is modified and the position of groups. The more the number of hydroxyl modified, the better the activity of the derivatives. When the position of groups on enantiomers is different, the activity of them is also different.In a word, modern pharmaceutical industry has become a major industry of the 21st century in China. So, the study of innovative drugs is of great social and economic benefit. We modified the kirenol and tested the inhibitory activity on nitric oxide (NO) production in LPS-activated macrophage. We have gotten some delectable result and established a good basis for further research of kirenol.更多还原