【作者】 王珊；

【导师】 黄胜阳；

【作者基本信息】 北京工业大学 ， 生物化学与分子生物学， 2012， 硕士

【摘要】 艾滋病又称获得性免疫缺陷综合症(Acquired Immunodeficiency Syndrome,AIDS)，是由人类免疫缺陷病毒(Human Immunodeficiency Virus, HIV)感染人体淋巴细胞等所致。自美国1981年诊断出首例艾滋病病人以来，艾滋病病毒在全球范围内的传播速度惊人，已有200余个国家和地区受到艾滋病的严重威胁。由于迄今仍无艾滋病疫苗问世，抗人类免疫缺陷病毒药物仍然是艾滋病治疗的主要手段。然而现有抗艾滋病药物大多是化学治疗药，具有易产生耐药性、毒副作用大及价格贵等缺点。因此众多学者把目光投向了具有化学结构多样性和生物活性的天然药物，从天然药物中开发研制作用于HIV生活周期新靶点的高效、无毒、价廉的药物已成为新世纪人们关注的热点。活血丹为中医临床常用中药，具有广泛的药理作用，如抗癌、止血、护肝、抗病毒、抗氧化、免疫调节等作用，并且近年来研究发现活血丹具有抗HIV的活性，其具体的活性成分和作用机理还有待进一步的研究。活血丹的主要化学成分是醌类化合物，并且从中还分离到一些其它类化合物如萜类、环肽类、多糖类等成分。研究认为，醌类化合物是活血丹植物中主要的药理活性成分。本研究以常用中药活血丹为研究对象，以HIV gp41为靶蛋白，通过采用BIA技术对其进行体外抗HIV试验，表明活血丹总提物具有一定的抗HIV作用，通过活性筛选初步确定氯仿萃取物、乙酸乙酯萃取物以及正丁醇萃取物都具有一定的抗HIV活性，应用硅胶柱吸附、葡聚糖凝胶柱、大孔吸附树脂柱、制备薄层等色谱分离方法及高效液相等现代分离纯化技术，从中提取分离得到18个单体化合物。采用核磁共振波谱、与对照品随同进行薄层层析、化学反应等方法，鉴定了其中16个化合物的结构，它们分别是：2,2-二甲基-5-甲氧羰基-6-羟基萘骈[1,2-b]吡喃、1-羟基-3-甲氧基-2-(2-氧代丙基)-9,10-蒽醌、1-羟基-3-(乙氧基-甲基)-9,10-蒽醌、β-谷甾醇、1,3-二羟基-2-(乙氧基-甲基)-9,10-蒽醌、2,2-二甲基-3,4-二氢-5-甲氧羰基-6-羟基萘骈[1,2-b]吡喃、4-羟基-3,5-二甲氧基苯甲酸、4-羟基-3-甲氧基-苯甲酸甲酯、1-羟基-2-乙氧羰基-9,10-蒽醌、4-(3-(2-甲氧基)-1,4-二氧-1,4-二氢萘酮)-1,3,5,10-呋喃酮-[2,3-c]蒽醌、1,3-二羟基-2-甲氧羰基-9,10-蒽醌、1,3-二羟基-8-甲氧基-2-甲基-9,10-蒽醌、1,3-二羟基-6-甲氧基-2-甲基-9,10-蒽醌、3-乙氧基苯甲酸、1-羟基-3-甲氧基-2-羧基-9,10-蒽醌、1-羟基-2-甲氧羰基-9,10-蒽醌，另外2个化合物的结构需要进一步的鉴定。其中化合物1-羟基-3-甲氧基-2-(2-氧代丙基)-9,10-蒽醌、1-羟基-3-(乙氧基-甲基)-9,10-蒽醌和3-乙氧基苯甲酸等3个化合物为首次从该中药中分离得到。本研究得到多种化学成分，填补了中药活血丹化学成分研究方面的空白，并为后续的单体活性研究工作奠定了较好的基础。更多还原

【Abstract】 AIDS, known as acquired immune deficiency syndrome, is caused by humanimmunodeficiency virus which infects human lymphocytes. Since the first case ofAIDS patients is diagnosed in1981, HIV spread worldwide at an alarming rate and isa serious threat to more than200countries and regions. As so far, due to the advent ofno AIDS vaccine, anti-human immunodeficiency virus drugs are still the primarymeans of AIDS treatment.However, the existing anti-AIDS drugs are most chemotherapy drugs which havesome disadvantages such as drug resistance, toxic-side effects, high price and so on.Many scholars turn their attention to the natural medicine because of its diversity ofchemical structure and biological activity. Developing efficient, non-toxic,inexpensive drugs which can target in the HIV life cycle from the natural medicinehas become a focus of attention in the new century.HUOXUEDAN is a clinical drug commonly used in Chinese medicine and has awide range of pharmacological effects such as anti-cancer, stop bleeding, liverprotection, anti-virus, antioxidant, immunomodulatory role; in recent years, studyfound that HUOXUEDAN has anti-HIV activity, its specific active ingredients andmechanism of action remains to have a further research. The main chemicalcomponents of HUOXUEDAN are quinones, terpenes,cyclic peptide, polysaccharidesand other ingredients are also isolated from HUOXUEDAN. Quinones arepharmacologically active ingredients of HUOXUEDAN.Through BIAcore research we find that the chloroform extraction, ethyl acetateextraction and butanol extraction possess anti-HIV activity. By using various chemicaland chromatographic methods, separation and purification work were performedsmoothly,we already got18compounds from HUOXUEDAN, and16of them wereidentified according to chemical methods, NMR data and references, they were:2,2-dimethyl-5-carbomethoxy-6-hydroxynaphtho[1,2-b]pyran;1-hydroxy-3-methoxy-2-(2-oxopropyl)-9,10-anthraquinone;1-hydroxyl-3-(ethoxy-methyl)-9,10-anthraquino-ne;β-sitosteol;1,3-dihydroxy-2-(ethoxymethyl)-9,10-anthraquinone;2,2-dimethyl-3,4-dihydro-5-carbomethoxy-6-hydroxynaphtho[1,2-b]pyran;4-hydroxy-3,5-dimethoxybenzoic acid;4-hydroxy-3-methoxy-benzoic acid methyl ester;1-hydroxy-2-ethoxy-carbonyl-9,10-anthraquinone;4-(3-(2-methoxypropan-2-yl)-1,4-dioxo-1,4-dihydronap-hthalen-2-yl)anthra[2,3-c]furan-1,3,5,10-tetraone;1,3-dihydroxy-2-methoxycarbonyl-9,10-anthraquinone;1,3-dihydroxy-8-methoxy-2-methyl-9,10-anthraquinone;1,3-dihyd-roxy-6-methoxy-2-methyl-9,10-anthraquinone;3-ethoxy benzoic acid;1-hydroxy-3-methoxy-2-carboxylic acid-9,10-anthraquinone;1-hydroxy-2-carbomethoxy-9,10-ant-hraquinone. Furthermore,1-hydroxy-3-methoxy-2-(2-oxopropyl)-9,10-anthraquinone; 1-hydroxy-3-(ethoxy-methyl)-9,10-anthraquinone and3-ethoxy benzoic acid areobtained from HUOXUEDAN for the first time. The other compounds are stillundergoing the process of structural elucidation. Above all, this project is a helpfulguide for the selection of bioactive compounds from HUOXUEDAN in the future.更多还原