天然药物中抗HIV活性成分研究进展

天然产物往往具有独特的化学结构,是发现新药或药物先导物的重要途径之一.目前,从天然药物中发现了多种结构类型的具有抗HIV活性的化合物,如多糖类、生物碱类、香豆素类、黄酮类、木脂素类、醌类、酚酸类、萜类等,并且其作用机制并不局限于抑制逆转录酶或蛋白酶,可在HIV复制周期的各个环节发挥作用,本文按天然产物的结构类型进行归纳,综述了近年来源自天然药物的抗HIV活性成分研究进展,总结了的151个具有抗HIV活性的天然产物结构及其活性测试数据,以期为从天然化学成分中寻找、发现并研究开发有效的抗艾滋病药物提供线索.     

黑老虎木脂素化学成分及药理作用研究进展CSCD

黑老虎是一种具有药用、食用、观赏等多种价值的经济植物。黑老虎的化学成分及药理作用研究进展较快,目前已分离出200余种化合物,其中木脂素类化合物121种,具有抗炎、抗肿瘤、抗HIV、抗凝血、抗氧化、保肝等多种药理作用。现对黑老虎中木脂素类化学成分和药理作用进行综述,为黑老虎的深入开发利用提供参考。     

五味子科药用植物亲缘学初探

五味子科Schisandraceae隶属于双子叶植物门木兰亚纲Magnoliidae八角目Illiciales, 全球分布约60种, 包括两个属: 五味子属Schisandra和南五味子属Kadsura, 间断分布于亚洲东南部和北美东南部。本文归纳了中国五味子科植物两大类活性成分--木脂素和三萜的分布规律、传统疗效和现代药理活性, 并对中国五味子科的药用植物亲缘学进行了初探。联苯环辛烯类木脂素(I)集中分布于五味子科植物, 可以被认为是五味子科植物的特征性化学成分, 除了传统的保肝作用外, 这类化合物中很多具有潜在的抗氧化、抗肿瘤和抗HIV活性, 一些联苯环辛烯类的木脂素, 尤其是在八元环C-6、C-9位上具有羟基或者酯化取代具有更好的抗HIV和抗肿瘤活性; 而螺苯骈呋喃型联苯环辛烯类木脂素(II)绝大多数存在于南五味子属, 其特殊的螺苯骈呋喃环及其钙拮抗、抗凝血和抑制血小板聚集的活性, 不仅初步说明了民间南五味子属药用植物藤茎具有较强活血化瘀药理作用的活性物质基础, 也提示在对南五味子属的药材质量标准研究中, 可以考虑以此类成分作为定性定量指标。五味子科植物中木脂素成分的分布规律提示, 在演化程度上五味子属植物较南五味子属植物更原始。环菠萝蜜烷类三萜在五味子属和南五味子属均有分布, 尤其是A环开环的环菠萝蜜烷类三萜(II)在抗HIV和抗肿瘤活性方面具有很好的潜力, 而结构更进化的7/7/5/6型三萜内酯(IV)显示了很强的细胞毒活性, 目前只在南五味子属的长梗南五味子K. longipedunculata中发现。从五味子属的小花五味子S. micrantha和狭叶五味子S. lancifolia中分离得到的多个成环复杂且高度氧化的类三萜内酯中也发现具有抗肿瘤和抗HIV潜力。     

杜仲皮中木脂素类化学成分及其对高糖诱导的肾小球系膜细胞保护作用研究

研究杜仲皮中的木脂素类化学成分,并评价其对高糖诱导的人肾小球系膜细胞的保护作用。将杜仲树皮粉碎后,纯化水提取,采用HPD-100大孔吸附树脂、硅胶、Sephadex LH-20、半制备高效液相色谱等方法进行分离,根据理化性质和波谱学方法鉴定化合物结构,从杜仲皮水提物中分离鉴定了6个木脂素类化合物,包括caruilignan D(1)、(-)-表松脂素(2)、(+)-松脂素4-O-β-D-吡喃葡萄糖苷(3)、(-)-松脂素4-O-β-D-吡喃葡萄糖苷(4)、(+)-中脂素4-O-β-D-吡喃葡萄糖苷(5)和(+)-中脂素4′-O-β-D-吡喃葡萄糖苷(6),化合物1为首次从杜仲中分离得到。在高糖诱导异常增殖的肾小球系膜细胞模型中,采用MTT法检测木脂素类化合物干预后的细胞活力。化合物5能剂量依赖性地抑制细胞增殖,对高糖诱导的肾小球系膜细胞具有保护活性。     

鬼臼毒素药源植物及其资源

鬼臼毒素属于木脂素类化合物中的芳基萘类,是一类重要的、具有一定的抗肿瘤和抗病毒活性的天然产物,能够产生鬼臼毒素类化合物的植物受到各国学者的关注。本文对近年来这类植物的形态、生境、化学成分及其内生菌代谢产物的研究进行了回顾。     

连翘中双四氢呋喃木脂素类化学成分及其波谱特征

本文主要研究了连翘中双四氢呋喃木脂素类化学成分的~(13)C NMR和CD谱的特征。通过大孔吸附树脂、硅胶、Sephadex LH-20、反相HPLC等多种色谱手段对连翘的50%乙醇提物中的木脂素类成分进行分离纯化,并根据理化性质和波谱数据鉴定其结构,最终分离得到了7个双四氢呋喃木脂素类化合物,分别为(+)-8-hydroxyepipinoresinol-4-O-β-D-glucopyranoside(1)、(+)-8-hydroxyepipinoresinol-4′-O-β-D-glucopyranoside(2)、(+)-松脂素-4-O-β-D-葡萄糖苷(3)、(+)-表松脂素-4-O-β-D-葡萄糖苷(4)、(+)-表松脂素-4′-O-β-D-葡萄糖苷(5)、(+)-phyllrin(6)和(+)-pinoresinol(7)。对这7个双四氢呋喃木脂素类化合物的~(13)C NMR和CD特点进行了总结。     

鸡脚参中一个新木脂素

从鸡脚参[Orthosiphon wulfenioides (Diels)Hand.-Marzz.]根中提取分离了一个新的木脂素,命名为鸡脚参木脂素。其结构由NMR、MS等波谱分析确定。初步活性实验显示该化合物不具备抗真菌和抗肿瘤活性。     

银胶菊中木脂素类成分的研究CSCD

研究银胶菊Parthenium hysterophorus Linn.中木脂素类化学成分。采用硅胶、Sephadex LH-20、MCI、ODS和半制备型HPLC等色谱技术,从银胶菊地上部分的95%乙醇提取物中分离得到5个木脂素类化合物(1~5),通过波谱学数据分析,将它们的结构分别鉴定为(7S,8S,7′E)-4,9,9′-三羟基-3,5,7,3′,5′-五甲氧基-8,4′-氧新木脂素-7′-烯(1)、丁香脂素(2)、杜仲树脂酚(3)、松脂素(4)、五味子醇甲(5)。其中,化合物1为新化合物,化合物3~5为首次从银胶菊中分离得到。对所有化合物进行了体外抗神经炎症活性筛选,结果显示,化合物1对LPS激活的BV-2小胶质细胞释放NO具有显著抑制作用,IC_(50)为21.10±1.97μmol/L,其余化合物均无活性,且所有化合物在测试浓度下均不影响细胞的存活率。     

芝麻素研究进展

芝麻素是存在于芝麻种子和芝麻油中的木脂素类化合物中的一种,已从多种植物中分离出来。芝麻素具有较强的抗氧化活性,在生物体内具有降低血清胆固醇、调节脂质代谢、稳定血压和抗癌等多种生理学活性。本文综述了芝麻素的天然来源、分离检测方法及其生理学活性等方面的研究现状。     

不同地理种源杜仲叶片中丁香脂素二糖甙和京尼平甙酸含量的分析

杜仲(Eucommia ulmoides Oliv.)为中国特有的重要经济树种.传统上取其皮入药,但有研究表明杜仲叶片中的化学成分与皮相同且药理作用相似[1].杜仲的皮叶中主要药用成分为环烯醚萜类和木脂素类等次生代谢产物[1].丁香脂素二糖甙(SGD)为木脂素类化合物,对磷酸二酯酶(CAMP)有很强的抑制活性[2];京尼平甙酸(GA)为环烯醚萜类化合物,也是杜仲皮叶中的主要成分之一,具有导泻[3]、抗高血压[4]以及预防性功能低下、增强记忆功能、抗癌、抗氧化、促进胆汁分泌等功能[1].有研究表明, 不同地理种源、不同季节的杜仲叶片中的活性成分的含量差异明显[5].因此,研究不同地理种源春、秋季杜仲叶片中丁香素二糖甙和京尼平甙酸的含量的差异,可以为杜仲的选择育种和叶片适时采收提供科学依据.     

1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide (ribavirin) and 5-ethynyl-1-beta-D-ribofuranosylimidazole-4-carboxamide (EICAR) markedly potentiate the inhibitory effect of 2',3'-dideoxyinosine on human immunodeficiency virus in peripheral blood lymphocytes.

Ribavirin and EICAR are two antiviral agents that share a similar antiviral activity spectrum and are targeted at inosine 5'-monophosphate (IMP) dehydrogenase. Neither ribavirin nor EICAR inhibit the replication of human immunodeficiency virus (HIV) in peripheral blood lymphocyte cells (PBL) at subtoxic concentrations. However, both compounds markedly potentiate the anti-HIV activity of 2',3'-dideoxyinosine (DDI) in PBL cells without a marked increase of toxicity. Both the increased IMP levels and the decreased guanine nucleotide levels caused by ribavirin and EICAR may be responsible for their potentiating effect on the anti-HIV activity of DDI.     

A study of the topoisomerase II activity in HIV-1 replication using the ferrocene derivatives as probes

Human Topoisomerase II is present in two isoforms, 170KDa alpha and 180KDa beta. Both the isoforms play a crucial role in maintenance of topological changes during DNA replication and recombination. It has been shown that Topoisomerase II activity is required for HIV-1 replication and the enzyme is phosphorylated during early time points of HIV-1 replication. In the present study, we have studied the molecular action of Topoisomerase II inhibitors, azalactone ferrocene (AzaFecp), Thiomorpholide amido methyl ferrocene (ThioFecp), and Ruthenium benzene amino pyridine (Ru(ben)Apy) on cell proliferation and also on various events of HIV-1 replication cycle. The Topoisomerase II beta over-expressing neuroblastoma cell line shows a higher sensitivity to these compounds compared to the Sup-T1 cell line. All the three Topoisomerase II inhibitors show significant anti-HIV activity at nanomolar concentrations against an Indian isolate of HIV-1(93IN101) in Sup-T1 cell line. An analysis of action of these compounds on proviral DNA synthesis at 5h of post-infection shows that they inhibit proviral DNA synthesis as well as the formation of pre-integration complexes completely. Further analysis, using polymerase chain reaction and western blot, showed that both the Topoisomerase II alpha and beta isoforms are present in the pre-integration complexes, suggesting their significant role in HIV-1 replication.     

Natural anti-HIV agents-part I: (+)-demethoxyepiexcelsin and verticillatol from Litsea verticillata

The eudesmane sesquiterpenoid, verticillatol (1), as well as the lignan, (+)-5'-demethoxyepiexcelsin (2), and a known lignan, (+)-epiexcelsin (3), were isolated from Litsea verticillata Hance. Lignan 2 showed moderate anti-HIV activity with an IC(50) value of 16.4 microg/ml (42.7 microM), while the known lignan 3 was inactive up to a concentration of 20 microg/ml (48.3 microM). Compound 1 demonstrated weak activity with an IC(50) value of 34.5 microg/ml (144.7 microM) while being devoid of cytotoxicity at 20 microg/ml. The structures were elucidated by 1D and 2D NMR spectroscopy, and the absolute configuration of the new sesquiterpenoid was determined by the generation of Mosher esters.     

Discovery of the first small molecule inhibitor of human DDX3 specifically designed to target the RNA binding site: towards the next generation HIV-1 inhibitors

Efficacy of currently approved anti-HIV drugs is hampered by mutations of the viral enzymes, leading invariably to drug resistance and chemotherapy failure. Recent data suggest that cellular co-factors also represent useful targets for anti-HIV therapy. Here we describe the identification of the first small molecules specifically designed to inhibit the HIV-1 replication by targeting the RNA binding site of the human DEAD-Box RNA helicase DDX3. Optimization of a easily synthetically accessible hit (1) identified by application of a high-throughput docking approach afforded the promising compounds 6 and 8 which proved to inhibit both the helicase and ATPase activity of DDX3 and to reduce the viral load of peripheral blood mononuclear cells (PBMC) infected with HIV-1.     

5-Alkyl-2-[(aryl and alkyloxylcarbonylmethyl)thio]-6-(1-naphthylmethyl) pyrimidin-4(3H)-ones as an unique HIV reverse transcriptase inhibitors of S-DABO series

The introduction of a beta-carbonyl group to the C-2 side chain of S-DABO led to the finding of a series of novel potent anti-HIV agent. Some derivatives proved to be highly effective in inhibiting HIV-1 replication at nanomolar concentrations. Furthermore, the novel S-DABOs differ from the classical NNRTIs in that some compounds are active against both HIV-1 and HIV-2. They might interfere with another target or at least act on RT in a different way as compared to typical NNRTIs.     

表达萤光素酶的HIV药物筛选细胞模型的建立

目的:构建基于萤光素酶的单次复制人免疫缺陷病毒(HIV)细胞模型,用于抗HIV药物的筛选。方法:构建含萤光素酶报告基因的假型慢病毒质粒,将疱疹性口炎病毒外膜糖蛋白(VSV-G)的表达质粒、HIV-1 Rev蛋白表达质粒、HIV Gag-Pol蛋白表达质粒和含萤光素酶报告基因的重组慢病毒质粒共转染HEK 293FT细胞,制备假型慢病毒;在假型慢病毒生产和再感染新鲜HEK 293FT细胞的过程中加入逆转录酶和蛋白酶抑制剂(如AZT),检测再感染的细胞中萤光素酶的表达水平,从而判断药物对HIV的抑制作用。结果:构建了含萤光素酶报告基因的重组慢病毒质粒pLenti-Luc;利用已知抗HIV药物AZT进行测试,发现HIV药物处理组细胞中萤光素酶活性远低于对照组。结论:建立了基于萤光素酶的HIV药物筛选细胞模型,该系统使用单次复制的报告病毒,具有良好的安全性,而使用萤光素酶基因作为报告基因使该系统具备极高的敏感性,该系统适合于进行高通量药物筛选。     

Anti-AIDS agents. Part 61: Anti-HIV activity of new podophyllotoxin derivatives

A series of novel podophyllotoxin derivatives containing structural modifications at C-4 (7-14), C-4' (16-17), and the methylenedioxy A-ring (23-28) was synthesized and tested for inhibition of HIV replication. Four of these compounds (25-28) were previously reported to show EC(50) values of <0.001 microg/mL and therapeutic index (TI) values >120. Three of the newly tested compounds (8, 12, and 20) showed good activity with EC(50) values of 0.012, <0.001, and 0.389 microg/mL and TI values of 19.1, >16, and 19.4, respectively. A comparison of the anti-HIV activity of these derivatives suggested that an opened A-ring with 6,7-dimethoxy substitution and a 4'-demethylated E ring enhanced anti-HIV activity.     

Discovery of novel low-molecular-weight HIV-1 inhibitors interacting with cyclophilin A using in silico screening and biological evaluations

Cyclophilin A has attracted attention recently as a new target of anti-human immunodeficiency virus type 1 (HIV-1) drugs. However, so far no drug against HIV-1 infection exhibiting this mechanism of action has been approved. To identify new potent candidates for inhibitors, we performed in silico screening of a commercial database of more than 1,300 drug-like compounds by using receptor-based docking studies. The candidates selected from docking studies were subsequently tested using biological assays to assess anti-HIV activities. As a result, two compounds were identified as the most active. Specifically, both exhibited anti-HIV activity against viral replication at a low concentration and relatively low cytotoxicity at the effective concentration inhibiting viral growth by 50 %. Further modification of these molecules may lead to the elucidation of potent inhibitors of HIV-1.