【作者】 姜冰；

【导师】 向文胜；

【作者基本信息】 东北农业大学 ， 生物化学与分子生物学， 2008， 硕士

【摘要】 从天然产物中筛选发现新型结构的活性化合物是天然产物研究的重点领域。传统的天然产物化学研究内容主要为通过萃取、层析以及色谱制备等手段进行分离纯化,获得达到一定纯度的单体化合物,测定其UV、IR、MS、NMR等谱学数据,确定化合物的结构;再进行体外或体内生物活性及作用机理研究。本文运用经典的植化研究方法对垂子买麻藤(Gnetum pendulum)中的化合物进行提取分离以及结构鉴定。应用萃取、正相硅胶柱层析、凝胶Sephadex LH-20柱层析多种分离手段,一共分离得到四个化合物,应用现代波谱技术鉴定了化合物的结构。最终确定来自此植物的四个化合物是:3,5-二羟基-4-甲氧基均二苯代乙烯(Ⅰ)、银松素(Ⅱ)、丹叶大黄素(Ⅲ)、射干乙素(Ⅳ)。其中化合物Ⅰ为首次报道的一全新的化合物。此外本研究以枯草芽孢杆菌(Bacillus subtilis)和藤黄微球菌(Sarcina lutea)为活性检测菌,利用活性跟踪的办法对一株放线菌发酵液的活性成分进行分离以及化合物的结构鉴定。应用正相硅胶柱层析、反相柱层析、凝胶Sephadex LH-20柱层析以及反相HPLC(High Performance Liquid Chromatography)制备等多种层析技术,一共分离得到七个化合物,应用现代波谱技术鉴定化合物的结构。这几个化合物分别为:8-氯-染料木素(1)、染料木素(2)、大豆苷元(3)、3’,4’,7-三羟基异黄酮(4)、芒柄花黄素(5)、5-羟基-5甲基-2-己烯酸(6)和腺苷(7)。其中化合物5-羟基-5-甲基-2-己烯酸(6)和腺苷(7)具有很好的抗菌活性。在分离得到的化合物中,发现8-氯-染料木素(1)可能是由染料木素(2)由放线菌经过生物氯化得到的,为了证实这一观点,对不同发酵期间两种物质的含量进行了研究,发现在发酵达到第三天时,染料木素大量减少,与此同时8-氯-染料木素的含量增加,说明此放线菌具有生物氯化的特性。更多还原

【Abstract】 Discovering new active natural products from natural resources is an important approach for new drug research and development. Traditional nature product chemistry is focused on isolation, purification and structure elucidation. The pure compound’s structure was determined by UV, IR, MS and NMR, its biological activity was tested by in vitro or in vivo experiment.In our investigation of chemical constituents from Gnetum Pendulum, four compounds were isolated by the methods of normal-phase silica gel and sephadex LH-20 chromatography. Then the compounds were elucidated on the basis of chemical and spectral methods. Four compounds were elucidated as 3, 5-dihydroxy-4-methoxystilbene (Ⅰ), pinosylvin (Ⅱ), rhapontigenin (Ⅲ) and shegansu B (Ⅳ), the compoundⅠis a new compound.Sarcina lutea (ATCC 9341) and Bacillus subtilis (ATCC 6633) were treated as bioassay-guided, they led to the isolation of the active constituents. Compounds have been isolation from the actinomycete by repeatedly chromatography on normal and reversed-phase silica gel columns, sephadex LH-20 and final purification by reversed preparative HPLC. By detailed analysis of the spectral data (IR, ESIMS, 1HNMR, 13CNMR, DEPT, COSY, NOESY, HMBC), seven of them were elucidated as following, 8-chloro-genistein(1), genistein(2), daidzein(3), 3’,4’,7-trihydroxy- isoflavone(4), formononetin(5), 5-hydroxy-5-methylhex-2-enoic acid(6) and adenosine(7). 5-hydroxy-5-methylhex-2-enoic acid(6) and adenosine(7) have good antibacterial activity.In the compounds isolated from the actinomycete, 8-chloro-genistein(1) was deduced the biological halogenation product of genistein, From the time course profile of this biotransformation, 8-chloro-genistein was observed from the third day of fermentation. The result completely confirmed that the actinomycete have biological halogenation characteristic.更多还原