奇壬醇衍生物的合成及对LPS诱导的巨噬细胞NO抑制活性的构效关系研究
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。
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.
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。
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.
引文
[1] Xiong J, Ma Y, Xu Y. Ditrepenoids from Siegesbeckia pubesecens [J].Phytochemistry, 1992;31(3):917-921.
[2] 中华人民共和国卫生部药典委员会.中华人民共和国药典[M].人民卫生出版社,2005 版,2005,255.
[3] 高松,李欣海,宋晓静等.豨莶草与金挖耳的鉴别[J],辽宁中医杂志,2004,28(11):959.
[4] Kanojia R.M. et al.J Org CK ahem,1982,47:1310
[5] 阴健,郭力弓.中药现代研究与临床应用[M].学苑出版社,1997,255-274.
[6] 吴巍,高辉,王丽君等.豨莶草化学成分与药理作用的研究进展,特产研究,2000,1:59-60.
[7] 宋立人.现代中药学大典[M].下册,人民卫生出版社,2001,2326.
[8] 张哲锋,杨连荣,李彦冰等.豨莶草化学成分与药理活性的研究进展,中药研究进展,2006,23(3):15
[9] Chan S L,Pallett A L,Morgan N G.Clotrimazole and efaroxan stimulate insulin secretion by different mechanisms in rat pancreatic islets.Naunyn Schmiedebergs Arch.Pharmacol, 1997,356:763-768.
[10] 徐叔云.药理实验方法学[M].第 3 版.人民卫生出版社,2002:901.
[11] 胡慧华, 汤鲁霞, 李小猛. 豨莶草生品和炮制品抗炎、抗风湿作用的实验研究[J]. 中国中药杂志, 2004, 29(6): 542-545.
[12] 刘寿山.中药研究文献摘要(1962～1974)[M].科学出版社,1979:747.
[13] Kang-BK.Inhibitory effects of Korean folk medicine HrChum on histamine release from mast cells in vivo and in vitro [J]. Ethnopharmacol,1997, 57(2):73.
[14] 中华本草,956-959.
[15] 王频萍.西北药学杂志,1990,5(2):39.
[16] 黄弘轩,李孝先,任国庆.豨莶草的舒血管作用[J].白求恩医科大学学报,1979,5(32):17-19.
[17] 有泽宗久.生药中 ACE 抑制活性成分的研究[J].国外医学·中药分册,1984,(6):31.
[18] 王浴生.中药药理与应用,人民卫生出版社,1983,1221-1222.
[19] Kim-HM.Effect of Siegesbeckia pubescen on immediate. Hypersensitivity [J]. Am.J chin.Med., 1997, 25(2): 163.
[20] 俞桂新,金岩敏,王峥涛等.豨莶草抗血栓有效组分筛选研究.上海中医药大学学报,2005,19(3):39.
[21] Kim-HM.Effect of Siegesbeckia pubescens on immediate hypersensitivity,Am J chinMed,1997,25(2):16.
[22] 蒋林.豨莶草注射液对实验性微循环障碍影响的观察〔J〕.中成药,1988,9:45.
[23] 王鹏.豨莶草乙醇提取物改善微循环及止痒的研究[J].Journal of Medical Forum, 2003, 24(12):19-21.
[24] 蒋 林 , 林 启 云 , 谢 金 鲜 . 豨 莶 草 药 理 实 验 研 究 [J]. 广 西 中 医药,1990,13(4):44.
[25] Dumas-Marc, the esters there of and oligo saccharides containing xylosen for improoing the functionality of epidermal cells [J].PCT Int Appl, 1999, 25:24009.
[26] 卜长武,杨正娟,那爱华.豨莶对小白鼠免疫功能的影响[J].中国中药杂志,1989,14(3): 44-45.
[27] 董祥英,陈敏,荆伟.毛梗豨莶生育活性化学成分的研究[J].药学学报,1989,24(11):833-836.
[86] 蒋华良,李松,陈凯先.药学科学前沿与发展方向.中国医药科技出版社,2000,137-147.
[29] Kanojia R.M. et al.J Org CK ahem,1982,47:1310.
[30] Min-Byung-Sun.Inhibitory activites of Korean plants on HIV-1protease,Nat P-Rod Sci,1998,4(4):241.
[31] 郭来,丁书文.复方豨莶草合剂抗动脉粥样硬化内皮细胞损伤实验研究[J].中医药学刊,2001,19(2):105-106.
[32] 王 达 一 .“ 九 制 豨 莶 丸 ” 治 疗 脑 血 管 意 外 后 遗 症 [J]. 江 苏 中医,1988,(12):21.
[33] 谭锦文等.湖北中医杂志,1984,(2):24.
[34] 赵力. 豨莶通栓丸治疗脑血栓的临床与实验研究[J].中国中西医结合杂志,1994,14(2):71.
[35] 湖北省卫生局.湖北中草药志[M].湖北科技出版社,1982,1061.
[36] 全国中草药新医疗法展览会资料选编(技术资料部分),1972,26.
[37] 彭世松.湖南医药杂志,1975,4:63.
[38] 枣庄矿物局官桥医院.枣庄医药,1976,7:60.
[39] 中医研究院针灸经络研究所等,新医药学杂志,1978,2:31.
[40] 黄育文.豨莶草可治鼻衄[J].新中医,1998,9:34.
[41] 唐净,宋京英.中国中医急症.2004,5:13.
[42] 张德文.豨钩地龙汤治子痫[J].浙江中医杂志,1991,7:317.
[43] 卜献春,廖金剑.豨莶通络液离子导入治疗糖尿病周围神经病变随机对照观察[J].现代康复,2001,5(7):33-34.
[44] 邓中伟.豨莶草治疗斑秃[J].中医杂志,2001,42(4):264.
[45] 隋冠华.豨莶草治疗药物性皮炎[J].中医杂志,2001,42(4):265.
[46] 钱瑞琴,张春英.豨莶草活性部位抗风湿作用机理研究[J].中国中西医结合杂志,2000,20(3):192.
[47]夏俊杰,朱晓鸣.运用豨莶草治疗痹证经验[J].山西中医,1997,13(2):7-8.
[48]钟开糠.豨莶草药用验方介绍,中国民族民间医药杂志.1988,2:13-14.
[49] 傅宏征,冯锐,楼之芩,毛梗豨莶的化学成分研究(I),中国药学杂志,1998,33(2): 140.
[50]傅宏征,冯锐,楼之芩,毛梗豨莶的化学成分研究(II),中国药学杂志,1998,5(33): 276.
[51] Baruah R N,Sharma R P,Madhusudanan K P,et al.1979,A New Melampolide from Siegesbeckia Orientalis, Phytochemistry, 18(6):991-994 .
[52] 果德安,张正高,叶国庆.豨莶脂溶性成分的研究,药学学报,1997,32(4):282-285.
[53] Kin JH,et al.phytochemistry,1979,18(5):894.
[54] Takao,et al.Tetrahedron Letter,1973,50:4991.
[55] Xiong Y,et al.Phytochemistry,1992,51(3):917.
[56] R.N.Baruah,R .P. Sharma,G .Thyagarajan,et al. Phytochemistry,1979,18:991-994.
[57] CA: 85,124182k.
[58] 傅宏征,李玉书,果德安等.中草药, 1999, 30:491-492.
[59] Bohlmann F,Uakupovic J,Zdero C.,et al,Neue Melampolide und Cis, Cis-Germactamolide aus Vertretern Der Subtribus, Melampodinae Phytochemistry, 1979, 18(4),625-630(Ger).
[60] K. Liu, E. Roder, Plant Med, 1991, 57:395-396.
[61] Qian R Q, Zhang C Y, Fu HZ. Study on therapeutic mechanism of anti-rheumatism action of Herbal Siegesbeckiae[J].Chin J Integrated Tradit Chin West Med,2000,20(3): 192-195.
[62] 信红岭,毕娟,刘淼等.奇壬醇的抗炎免疫实验研究,中草药,2005,36(6):866-867.
[63] 田志敏.非甾体抗炎药物及其制剂的研究进展,实用中西医结合临床,2007,7(1):92-93.
[64] Vane J.The evolution of non-steroidal anti-inflammatory drugs and their mechanisms of action[J].Drugs,1987,33(1):18-27.
[65] Moncada S,Higgs A,Fuchgott FR.International union of pharmacology nomenclature in nitric oxide research, Pharmacology Rev,1997,49:137-142.
[66] Curtis MJ,Pabla R.Nitric oxide supplementation or synthesis block-which is better approach for heart failure.Trends Pharmacol Sci,1997,18:239-243.
[67] 朱华,邹峥.非甾体抗炎药临床作用及副作用研究新进展,江西医药,2007,42(3):268-269.
[68] Marletta MA.Nitric oxide synthase structure and mechanism.J Biol Chem,1993,268:12231-12234.
[69] Singh G,Rosen D.NSAID-induced gastrointestinal complications [J].The Aramis Perspective, 1997, 25(51):8
[70] 林文翰.奇壬醇抗风湿性关节炎中药的新药开发研究.第七届北京生物医药产业发展论坛,2003.
[71] ZHU Xing Zu. Achievement on Pharmacology [M ]. Peop le’s Health Publishing House, 1997, 19- 29.
[72] Ialenti A,Moncada S,et al,Mondulation of acute inflammation by endogenous nitric oxide,Eur J Pharmacol,1992,211:177.
[73] Iuvone T,Carnuccio R,Di Rosa M,Modulation of granuloma ofrmation by endogenous nitric oxide, Eur J Pharmacol, 1994,265(1):89.
[74] Nussler A K, Di Silvio M , Billiar T R, et al. Stimulation of the nitric oxide synthase pathway in human hepatocytes by cytokines and endotoxin [ J ]. J Exp M ed,1992, 176 (1) : 261- 264.
[75] Southam E, Garthwaite J. The nitric oxide-cyclic GMP signalling pathway in rat brain [ J ]. N europharma-cology, 1993, 32: 1267.
[76] De Vente J. NOS and cGMP synthesis in the rat brain [J ]. Neuro science, 1998, 87 (1) : 207- 241.
[77] M urad F. Signal transduction using nitric oxide and cyclic guanosine monophosphate [ J ]. JAMA , 1996,276 (14) : 1189- 1192.
[78] Murad F. Discovery of some of the biological effects of nitric oxide and its ro le in cell signaling [ J ]. Angew Chem Int Ed, 1999, 38: 185- 186.
[79] 李甘地,病理学,人民卫生出版社,第 l 版,2001,81-82.
[80] Mayer B, Pfeiffer S, Schrammel A , et al. A new pathway of nitric oxide cyclic GMP signaling involving nitro soglutathione [J ]. J Biol Chem, 1998, 273 (6) : 3264- 3270.
[81] Skomsvoll JF, Wallenius M, Koksvik HS, et al. Drug insight: Anti-tumor necrosis factor therapy for inflammatory arthropathies during reproduction, pregnancy and lactation [J]. Nat Clin Pract Rheumatol, 2007, 3: 156-164.
[82] Mo XR, Luo XJ. Progress of studies on the mechanism of rheumatoid arthritis. Clin Res, 2007, 2: 336-340.
[83] Van der Helm-van Mil AH, Kurreeman FA, Toes RE, et al. Association of tumor necrosis factor alpha polymorphism and radiographic progression in rheumatoid arthritis [J]. Arthritis Rheum,2007,3: 1032-1033.
[84] Stefanovic-Racic M, Stadler J, Evans CH. Nitric oxide and arthritis [J]. Arthritis Rheum, 1993, 8: 1036-1044.
[85] Stefanovic-Racic M, Meyers K, Meschter C, et al. N-monomethyl arginine, an inhibitor of nitric oxide synthase, suppesses the development of adjuvant arthritis in rats [J]. Arthritis Rheum, 1994, 7: 1062-1069.
[86] 梁万年.关节炎-类风湿性关节炎[J],继续医学教育,2005,19:15-17.
[87] 黄文龙,彭司勋,张惠斌.药物化学进展.中国医药科技出版社,2000,81-95.
[88] Ginnings,P.;Baum,R.Aqueous Solubilities of the Isomeric Pentanols.J.Amer.Chem. Soc.1937,59:1111-1113.
[89] Harms,A.F,Nauta,W.T. The Effects of Alkyl Substitution in Drugs-Ⅰ. Substituted Dimethylaminothyl Benzhydryl Ethers,Med. Pharm.Chem, 1960,2:57-77.
[90] Walsmith J,Roubenoff R.Cachexia in rheumatoid arthritis[J],Int J Cardiol, 2002, 85: 89-99.
[91] 许文亮,郭新贵,徐延路. 冠心病患者血浆胆固醇与内皮素- C-反 应蛋白及血小板可溶性P选择素相关性研究. 新医学 2006,25(6): 330-331.
[92] Fischer J,Gere A.Timing of analog research in the medicinal chemistry,Pharmazie,2001,56:675-682.
[93] Sternbach L H, The Benzodiazepine Story, J.Med.Chem, 1979,22:1-7.
[94] Hamilton T C,Weir S W,Weston A H.Comparison of the Effects of BRI,34915 and Verapamil on Electrical and Mechanical Activity in Rat Portal Vein, Brit.J.Pharmacol, 1986,88:103-111.
[95] Evans J M, Fake C S, Hamilton T C, et al. Synthesis and Antihypertensive Activity of Substituted trans-4-Amino-3,4-dihydro-2,2-dimethyl-2H-1-benzopyran-3-ols, J.Med.Chem,1983,27,1127-1131.
[96] Black J W,Duncan W A M,Durant J C,et al.Definition and Antagonism of Histamine H2-Receptors, Nature,1972,236:385-390.
[97] Leonard D M.Ras Farnesyltransferase:A New Therapeutic Target,J.Med.Chem, 1997,40:2971-2990.
[98] Newman DJ, Cragg GM, Snader KM. Natural product as sources of new drugs over the period 1981-2002[J].J Nat Prod, 2003,66(1): 1022-1037.
[99] Reddy M V R,Rao M R,Rhodes D,Hansen M S T,et al.Lamellarin a 20-Sulfate,an Inhibitor of HIV-I Integrase Active against HIV-I Virus in Cell Culture, J .Med.Chem, 1999,42:1901-1907.
[100] Song Y,Connor D T,Sercel A D,et al. Synthesis, Structure-Activity Relationships and in Vivo Evaluations of Substituted Di-tert-butylphenols as a Novel Class of Potent, Selective, and Orally Active Cyclooxygenase-2 Inhibitors 2,1,3,4 and 1,2,4-Thiadiazole Series,J.Med.Chem,1999,42:1161-1169.
[101] Thorpe,D S,Edith Chan A W,Binnie A,et al.Efficient discovery of inhibitory ligands for diverse targets from a small combinatorial cbemical library of chimeric molecules, Biochem.Biophys.Res.Commun, 1999,266:62-65.
[102] Lahana R.How many leads from HTS?Evaluation of Diverse Compound Subsets from Chemical Structure Databases, J.Med.Chem,1999,41:478-488.
[103] Wenter J C.The sequence of the human genome,Science, 2001,291:1304-1335.
[104] Abagyan R,Totrov M.High-throughput docking and lead generatration, Curr. Opin.Struct.Biol,2000,5:375-382.
[105] Gohlke H.Statistical potentials and scoring functions applied to protein-ligand binding, Curr.Opin.Struct.Biol, 2001,11:231-235.
[106] 卡米尔·乔治·维尔特,创新药物化学,世界图书出版公司,2005,87.
[107] Chan S L,Pallett A L,Morgan N G.Clotrimazole and efaroxan stimulateinsulin secretion by different mechanisms in rat pancreatic islets.Naunyn Schmiedebergs Arch.Pharmacol, 1997,356:763-768.
[108] Chan S L,Pallett A L,Morgan N G.The effect of the putative endogenous imidazoline recetor ligand, clonidine-displacing substances, on insulin secretion from rat and human islets of Langerbans, Br.J.Pharmacol,1997,120:926-932.
[109] 郑虎.药物化学,2003,第 4 版,284
[110] Farrell AJ, Blake DR, Palmer RM, et al. Increased concentrations of nitrite in synovial fluid and serum samples suggest increased nitric oxide synthesis in rheumatic diseases [J]. Ann Rheum Dis, 1992, 11: 1219-1222.
[111] Lee HJ, Kim NY, Jang MK, et al. A sesquiterpene, dehydrocostus lactone, inhibits the expression of inducible nitric oxide synthase and TNF-alpha in LPS-activated macrophages [J]. Planta Med, 1999, 2: 104-108.
[112] 于 德 泉 . 从 天 然 产 物 创 制 新 药 的 趋 势 [J]. 中 国 工 程 科 学 , 1999,1(1):87-90.
[2] 中华人民共和国卫生部药典委员会.中华人民共和国药典[M].人民卫生出版社,2005 版,2005,255.
[3] 高松,李欣海,宋晓静等.豨莶草与金挖耳的鉴别[J],辽宁中医杂志,2004,28(11):959.
[4] Kanojia R.M. et al.J Org CK ahem,1982,47:1310
[5] 阴健,郭力弓.中药现代研究与临床应用[M].学苑出版社,1997,255-274.
[6] 吴巍,高辉,王丽君等.豨莶草化学成分与药理作用的研究进展,特产研究,2000,1:59-60.
[7] 宋立人.现代中药学大典[M].下册,人民卫生出版社,2001,2326.
[8] 张哲锋,杨连荣,李彦冰等.豨莶草化学成分与药理活性的研究进展,中药研究进展,2006,23(3):15
[9] Chan S L,Pallett A L,Morgan N G.Clotrimazole and efaroxan stimulate insulin secretion by different mechanisms in rat pancreatic islets.Naunyn Schmiedebergs Arch.Pharmacol, 1997,356:763-768.
[10] 徐叔云.药理实验方法学[M].第 3 版.人民卫生出版社,2002:901.
[11] 胡慧华, 汤鲁霞, 李小猛. 豨莶草生品和炮制品抗炎、抗风湿作用的实验研究[J]. 中国中药杂志, 2004, 29(6): 542-545.
[12] 刘寿山.中药研究文献摘要(1962～1974)[M].科学出版社,1979:747.
[13] Kang-BK.Inhibitory effects of Korean folk medicine HrChum on histamine release from mast cells in vivo and in vitro [J]. Ethnopharmacol,1997, 57(2):73.
[14] 中华本草,956-959.
[15] 王频萍.西北药学杂志,1990,5(2):39.
[16] 黄弘轩,李孝先,任国庆.豨莶草的舒血管作用[J].白求恩医科大学学报,1979,5(32):17-19.
[17] 有泽宗久.生药中 ACE 抑制活性成分的研究[J].国外医学·中药分册,1984,(6):31.
[18] 王浴生.中药药理与应用,人民卫生出版社,1983,1221-1222.
[19] Kim-HM.Effect of Siegesbeckia pubescen on immediate. Hypersensitivity [J]. Am.J chin.Med., 1997, 25(2): 163.
[20] 俞桂新,金岩敏,王峥涛等.豨莶草抗血栓有效组分筛选研究.上海中医药大学学报,2005,19(3):39.
[21] Kim-HM.Effect of Siegesbeckia pubescens on immediate hypersensitivity,Am J chinMed,1997,25(2):16.
[22] 蒋林.豨莶草注射液对实验性微循环障碍影响的观察〔J〕.中成药,1988,9:45.
[23] 王鹏.豨莶草乙醇提取物改善微循环及止痒的研究[J].Journal of Medical Forum, 2003, 24(12):19-21.
[24] 蒋 林 , 林 启 云 , 谢 金 鲜 . 豨 莶 草 药 理 实 验 研 究 [J]. 广 西 中 医药,1990,13(4):44.
[25] Dumas-Marc, the esters there of and oligo saccharides containing xylosen for improoing the functionality of epidermal cells [J].PCT Int Appl, 1999, 25:24009.
[26] 卜长武,杨正娟,那爱华.豨莶对小白鼠免疫功能的影响[J].中国中药杂志,1989,14(3): 44-45.
[27] 董祥英,陈敏,荆伟.毛梗豨莶生育活性化学成分的研究[J].药学学报,1989,24(11):833-836.
[86] 蒋华良,李松,陈凯先.药学科学前沿与发展方向.中国医药科技出版社,2000,137-147.
[29] Kanojia R.M. et al.J Org CK ahem,1982,47:1310.
[30] Min-Byung-Sun.Inhibitory activites of Korean plants on HIV-1protease,Nat P-Rod Sci,1998,4(4):241.
[31] 郭来,丁书文.复方豨莶草合剂抗动脉粥样硬化内皮细胞损伤实验研究[J].中医药学刊,2001,19(2):105-106.
[32] 王 达 一 .“ 九 制 豨 莶 丸 ” 治 疗 脑 血 管 意 外 后 遗 症 [J]. 江 苏 中医,1988,(12):21.
[33] 谭锦文等.湖北中医杂志,1984,(2):24.
[34] 赵力. 豨莶通栓丸治疗脑血栓的临床与实验研究[J].中国中西医结合杂志,1994,14(2):71.
[35] 湖北省卫生局.湖北中草药志[M].湖北科技出版社,1982,1061.
[36] 全国中草药新医疗法展览会资料选编(技术资料部分),1972,26.
[37] 彭世松.湖南医药杂志,1975,4:63.
[38] 枣庄矿物局官桥医院.枣庄医药,1976,7:60.
[39] 中医研究院针灸经络研究所等,新医药学杂志,1978,2:31.
[40] 黄育文.豨莶草可治鼻衄[J].新中医,1998,9:34.
[41] 唐净,宋京英.中国中医急症.2004,5:13.
[42] 张德文.豨钩地龙汤治子痫[J].浙江中医杂志,1991,7:317.
[43] 卜献春,廖金剑.豨莶通络液离子导入治疗糖尿病周围神经病变随机对照观察[J].现代康复,2001,5(7):33-34.
[44] 邓中伟.豨莶草治疗斑秃[J].中医杂志,2001,42(4):264.
[45] 隋冠华.豨莶草治疗药物性皮炎[J].中医杂志,2001,42(4):265.
[46] 钱瑞琴,张春英.豨莶草活性部位抗风湿作用机理研究[J].中国中西医结合杂志,2000,20(3):192.
[47]夏俊杰,朱晓鸣.运用豨莶草治疗痹证经验[J].山西中医,1997,13(2):7-8.
[48]钟开糠.豨莶草药用验方介绍,中国民族民间医药杂志.1988,2:13-14.
[49] 傅宏征,冯锐,楼之芩,毛梗豨莶的化学成分研究(I),中国药学杂志,1998,33(2): 140.
[50]傅宏征,冯锐,楼之芩,毛梗豨莶的化学成分研究(II),中国药学杂志,1998,5(33): 276.
[51] Baruah R N,Sharma R P,Madhusudanan K P,et al.1979,A New Melampolide from Siegesbeckia Orientalis, Phytochemistry, 18(6):991-994 .
[52] 果德安,张正高,叶国庆.豨莶脂溶性成分的研究,药学学报,1997,32(4):282-285.
[53] Kin JH,et al.phytochemistry,1979,18(5):894.
[54] Takao,et al.Tetrahedron Letter,1973,50:4991.
[55] Xiong Y,et al.Phytochemistry,1992,51(3):917.
[56] R.N.Baruah,R .P. Sharma,G .Thyagarajan,et al. Phytochemistry,1979,18:991-994.
[57] CA: 85,124182k.
[58] 傅宏征,李玉书,果德安等.中草药, 1999, 30:491-492.
[59] Bohlmann F,Uakupovic J,Zdero C.,et al,Neue Melampolide und Cis, Cis-Germactamolide aus Vertretern Der Subtribus, Melampodinae Phytochemistry, 1979, 18(4),625-630(Ger).
[60] K. Liu, E. Roder, Plant Med, 1991, 57:395-396.
[61] Qian R Q, Zhang C Y, Fu HZ. Study on therapeutic mechanism of anti-rheumatism action of Herbal Siegesbeckiae[J].Chin J Integrated Tradit Chin West Med,2000,20(3): 192-195.
[62] 信红岭,毕娟,刘淼等.奇壬醇的抗炎免疫实验研究,中草药,2005,36(6):866-867.
[63] 田志敏.非甾体抗炎药物及其制剂的研究进展,实用中西医结合临床,2007,7(1):92-93.
[64] Vane J.The evolution of non-steroidal anti-inflammatory drugs and their mechanisms of action[J].Drugs,1987,33(1):18-27.
[65] Moncada S,Higgs A,Fuchgott FR.International union of pharmacology nomenclature in nitric oxide research, Pharmacology Rev,1997,49:137-142.
[66] Curtis MJ,Pabla R.Nitric oxide supplementation or synthesis block-which is better approach for heart failure.Trends Pharmacol Sci,1997,18:239-243.
[67] 朱华,邹峥.非甾体抗炎药临床作用及副作用研究新进展,江西医药,2007,42(3):268-269.
[68] Marletta MA.Nitric oxide synthase structure and mechanism.J Biol Chem,1993,268:12231-12234.
[69] Singh G,Rosen D.NSAID-induced gastrointestinal complications [J].The Aramis Perspective, 1997, 25(51):8
[70] 林文翰.奇壬醇抗风湿性关节炎中药的新药开发研究.第七届北京生物医药产业发展论坛,2003.
[71] ZHU Xing Zu. Achievement on Pharmacology [M ]. Peop le’s Health Publishing House, 1997, 19- 29.
[72] Ialenti A,Moncada S,et al,Mondulation of acute inflammation by endogenous nitric oxide,Eur J Pharmacol,1992,211:177.
[73] Iuvone T,Carnuccio R,Di Rosa M,Modulation of granuloma ofrmation by endogenous nitric oxide, Eur J Pharmacol, 1994,265(1):89.
[74] Nussler A K, Di Silvio M , Billiar T R, et al. Stimulation of the nitric oxide synthase pathway in human hepatocytes by cytokines and endotoxin [ J ]. J Exp M ed,1992, 176 (1) : 261- 264.
[75] Southam E, Garthwaite J. The nitric oxide-cyclic GMP signalling pathway in rat brain [ J ]. N europharma-cology, 1993, 32: 1267.
[76] De Vente J. NOS and cGMP synthesis in the rat brain [J ]. Neuro science, 1998, 87 (1) : 207- 241.
[77] M urad F. Signal transduction using nitric oxide and cyclic guanosine monophosphate [ J ]. JAMA , 1996,276 (14) : 1189- 1192.
[78] Murad F. Discovery of some of the biological effects of nitric oxide and its ro le in cell signaling [ J ]. Angew Chem Int Ed, 1999, 38: 185- 186.
[79] 李甘地,病理学,人民卫生出版社,第 l 版,2001,81-82.
[80] Mayer B, Pfeiffer S, Schrammel A , et al. A new pathway of nitric oxide cyclic GMP signaling involving nitro soglutathione [J ]. J Biol Chem, 1998, 273 (6) : 3264- 3270.
[81] Skomsvoll JF, Wallenius M, Koksvik HS, et al. Drug insight: Anti-tumor necrosis factor therapy for inflammatory arthropathies during reproduction, pregnancy and lactation [J]. Nat Clin Pract Rheumatol, 2007, 3: 156-164.
[82] Mo XR, Luo XJ. Progress of studies on the mechanism of rheumatoid arthritis. Clin Res, 2007, 2: 336-340.
[83] Van der Helm-van Mil AH, Kurreeman FA, Toes RE, et al. Association of tumor necrosis factor alpha polymorphism and radiographic progression in rheumatoid arthritis [J]. Arthritis Rheum,2007,3: 1032-1033.
[84] Stefanovic-Racic M, Stadler J, Evans CH. Nitric oxide and arthritis [J]. Arthritis Rheum, 1993, 8: 1036-1044.
[85] Stefanovic-Racic M, Meyers K, Meschter C, et al. N-monomethyl arginine, an inhibitor of nitric oxide synthase, suppesses the development of adjuvant arthritis in rats [J]. Arthritis Rheum, 1994, 7: 1062-1069.
[86] 梁万年.关节炎-类风湿性关节炎[J],继续医学教育,2005,19:15-17.
[87] 黄文龙,彭司勋,张惠斌.药物化学进展.中国医药科技出版社,2000,81-95.
[88] Ginnings,P.;Baum,R.Aqueous Solubilities of the Isomeric Pentanols.J.Amer.Chem. Soc.1937,59:1111-1113.
[89] Harms,A.F,Nauta,W.T. The Effects of Alkyl Substitution in Drugs-Ⅰ. Substituted Dimethylaminothyl Benzhydryl Ethers,Med. Pharm.Chem, 1960,2:57-77.
[90] Walsmith J,Roubenoff R.Cachexia in rheumatoid arthritis[J],Int J Cardiol, 2002, 85: 89-99.
[91] 许文亮,郭新贵,徐延路. 冠心病患者血浆胆固醇与内皮素- C-反 应蛋白及血小板可溶性P选择素相关性研究. 新医学 2006,25(6): 330-331.
[92] Fischer J,Gere A.Timing of analog research in the medicinal chemistry,Pharmazie,2001,56:675-682.
[93] Sternbach L H, The Benzodiazepine Story, J.Med.Chem, 1979,22:1-7.
[94] Hamilton T C,Weir S W,Weston A H.Comparison of the Effects of BRI,34915 and Verapamil on Electrical and Mechanical Activity in Rat Portal Vein, Brit.J.Pharmacol, 1986,88:103-111.
[95] Evans J M, Fake C S, Hamilton T C, et al. Synthesis and Antihypertensive Activity of Substituted trans-4-Amino-3,4-dihydro-2,2-dimethyl-2H-1-benzopyran-3-ols, J.Med.Chem,1983,27,1127-1131.
[96] Black J W,Duncan W A M,Durant J C,et al.Definition and Antagonism of Histamine H2-Receptors, Nature,1972,236:385-390.
[97] Leonard D M.Ras Farnesyltransferase:A New Therapeutic Target,J.Med.Chem, 1997,40:2971-2990.
[98] Newman DJ, Cragg GM, Snader KM. Natural product as sources of new drugs over the period 1981-2002[J].J Nat Prod, 2003,66(1): 1022-1037.
[99] Reddy M V R,Rao M R,Rhodes D,Hansen M S T,et al.Lamellarin a 20-Sulfate,an Inhibitor of HIV-I Integrase Active against HIV-I Virus in Cell Culture, J .Med.Chem, 1999,42:1901-1907.
[100] Song Y,Connor D T,Sercel A D,et al. Synthesis, Structure-Activity Relationships and in Vivo Evaluations of Substituted Di-tert-butylphenols as a Novel Class of Potent, Selective, and Orally Active Cyclooxygenase-2 Inhibitors 2,1,3,4 and 1,2,4-Thiadiazole Series,J.Med.Chem,1999,42:1161-1169.
[101] Thorpe,D S,Edith Chan A W,Binnie A,et al.Efficient discovery of inhibitory ligands for diverse targets from a small combinatorial cbemical library of chimeric molecules, Biochem.Biophys.Res.Commun, 1999,266:62-65.
[102] Lahana R.How many leads from HTS?Evaluation of Diverse Compound Subsets from Chemical Structure Databases, J.Med.Chem,1999,41:478-488.
[103] Wenter J C.The sequence of the human genome,Science, 2001,291:1304-1335.
[104] Abagyan R,Totrov M.High-throughput docking and lead generatration, Curr. Opin.Struct.Biol,2000,5:375-382.
[105] Gohlke H.Statistical potentials and scoring functions applied to protein-ligand binding, Curr.Opin.Struct.Biol, 2001,11:231-235.
[106] 卡米尔·乔治·维尔特,创新药物化学,世界图书出版公司,2005,87.
[107] Chan S L,Pallett A L,Morgan N G.Clotrimazole and efaroxan stimulateinsulin secretion by different mechanisms in rat pancreatic islets.Naunyn Schmiedebergs Arch.Pharmacol, 1997,356:763-768.
[108] Chan S L,Pallett A L,Morgan N G.The effect of the putative endogenous imidazoline recetor ligand, clonidine-displacing substances, on insulin secretion from rat and human islets of Langerbans, Br.J.Pharmacol,1997,120:926-932.
[109] 郑虎.药物化学,2003,第 4 版,284
[110] Farrell AJ, Blake DR, Palmer RM, et al. Increased concentrations of nitrite in synovial fluid and serum samples suggest increased nitric oxide synthesis in rheumatic diseases [J]. Ann Rheum Dis, 1992, 11: 1219-1222.
[111] Lee HJ, Kim NY, Jang MK, et al. A sesquiterpene, dehydrocostus lactone, inhibits the expression of inducible nitric oxide synthase and TNF-alpha in LPS-activated macrophages [J]. Planta Med, 1999, 2: 104-108.
[112] 于 德 泉 . 从 天 然 产 物 创 制 新 药 的 趋 势 [J]. 中 国 工 程 科 学 , 1999,1(1):87-90.