茅苍术化学成分及药理活性研究

发布时间：2018-05-12 02:11

  本文选题：茅苍术 + 化学成分　； 参考：《北京协和医学院》2017年博士论文

【摘要】：茅苍术(Atractylodeslancea)为中医临床常用中药。为进一步阐明其药效物质基础,本文采用体外活性与紫外导向的方法对有效部位进行系统分离。采用多种柱色谱技术从正丁醇部位30%亚部位中共分离得到126个单体化合物,通过UV,IR,HRESIMS,1D和2DNMR等谱学技术鉴定出74个新化合物与52个已知化合物,并借助实验和理论ECD、Mo2(OAc)4和Rh2(OCOCF3)4诱导ECD以及X-ray等手段对新化合物的立体构型进行了确定。74个新化合物包括8个香根螺烷型倍半萜(1-8)、10个桉叶烷型倍半萜(9-18)、3个愈创木烷型倍半萜(19-21)、2个艾里莫酚烷型倍半萜(22,23)、11个链状C10-烯炔(24-34)、21个链状C13-烯炔(35-55)、6个链状C14-烯炔(56-61)、2个呋喃环多烯炔(62.63)、5个噻吩环多烯炔(64-68)、2个双四氢呋喃类木脂素(69,70)、2个酚苷(71,72)、1个吡嗪类生物碱(73)以及1个苯并四氢呋喃类衍生物(74)。解析出的已知化合物(75-126)主要包括木脂素、多烯炔、倍半萜以及香豆素等类型化合物。在化学成分研究的基础上,对得到的81个单体化合物进行了多方面的体外药理活性筛选。化合物2,4,22,24,27,71,72.73,84,109,122在10μM浓度下与扑热息痛合用时,对后者引起的肝细胞损伤有较好的保护作用,与模型组相比有统计学差异。化合物3,4,9,47,90,120,122,123在10μM浓度下对脂多糖诱导小胶质细胞释放的NO具有较好的抑制作用。本课题在开展过程中,还建立了一种能够简便、快速测定具有链状邻二醇片段的多烯炔苷相对构型(赤或苏)的1H NMR方法。选择氘代醋酸和重水作为测试溶剂,当链状邻二醇片段的3JHH偶合常数约为7.0 Hz时,则该化合物可归属为苏式构型,而当3JHH偶合常数约为3.5 Hz时,则该化合物可归属为赤式构型。溶液中氘代醋酸的比例可根据待测样品的实际溶解度进行微调。此种方法不需要对烯炔苷类化合物进行水解,极大缩减了实验的工作量,继而促进了类似化合物绝对构型的确定。此外,在确定香根螺烷倍半萜(1-5)绝对构型的过程中,还发现苷元中具有1个手性中心的化合物(1,2)与苷元中具有2个手性中心的化合物(3-5)呈现出相似的ECD谱图。借助计算ECD与X-单晶衍射技术,进一步研究发现此类结构中远离共轭体系的C-7位在240～260nm附近呈现较强的Cotton效应且强于C-5位的Cotton效应;化合物3-5在250 nm附近的Cotton效应可能为C-7与C-5位发色团Cotton效应的加权叠加。相关结果为香根螺烷型倍半萜立体构型的研究提供了参考。
[Abstract]:Atractylodeslancea (Atractylodeslancea) is a traditional Chinese medicine. In order to further elucidate the substance basis of its pharmacodynamics, the active parts were systematically separated by in vitro activity and UV-guided method. A total of 126 monomers were isolated from the 30% subfraction of n-butanol by various column chromatography techniques. 74 new compounds and 52 known compounds were identified by UVX IR HRESIMS1D and 2DNMR techniques. The stereotypes of the new compounds were determined by means of experimental and theoretical methods such as ECD-Mo2OAc4, Rh2(OCOCF3)4 induced ECD and X-ray. 74 new compounds include 8 Gyabgon spiroalkyl sesquiterpenes 1-8, 10 eucalyptane sesquiterpenes 9-18, and 3 guaiacane-type ploidy. Sesquiterpenoids 19-21, 2 alimohexane sesquiterpenes 2223C, 11 chain C10-enyne 24-34, 21 chain C13-enyne 35-55, 6 chain C14-enyne 56-61, 2 furancyclic polyenyne 62.63, 5 thiophene polyenyne 64-68, 2 dihydrofuran 697070, 2 pyrazine 71- 72o, 1 pyrazine Alkaloid 73) and one benzo-tetrahydrofuran derivative. The known compounds include lignin, polyenyne, sesquiterpene and coumarin. Based on the study of chemical constituents, 81 monomers were screened for their pharmacological activities in vitro. When compound 2H _ 4N _ 2H _ (22) O _ (22) C _ (27) 71C _ (72.73N) ~ (84) 109122 was combined with paracetamol at a concentration of 10 渭 M, it had a better protective effect on hepatocyte injury induced by paracetamol, which was significantly different from that in the model group. Compound 3H _ 4H _ 4N _ (47) C _ (901) 201 22123 had a good inhibitory effect on no release from microglia induced by lipopolysaccharide at a concentration of 10 渭 M. In the course of this study, a 1H NMR method was developed for the rapid determination of the relative configuration of polyenylethynine with chain O-diol fragments. Deuterated acetic acid and heavy water were selected as the test solvent. When the 3JHH coupling constant of the chain O-diol fragment was about 7.0Hz, the compound could be classified as a Soviet configuration, and if the 3JHH coupling constant was about 3.5Hz, the compound could be classified as a red-type configuration. The ratio of deuterated acetic acid in the solution can be fine-tuned according to the actual solubility of the sample to be tested. This method does not need to hydrolyze the alkynylate compounds, which greatly reduces the experimental workload and promotes the determination of the absolute configuration of similar compounds. In addition, during the determination of the absolute configuration of Gyabgon spirosine sesquiterpene 1-5), it was also found that the ECD spectra of the compounds with one chiral center in the glycoside and two chiral centers in the glycogen were similar to those in the glycogen. By calculating the ECD and X- single crystal diffraction technique, it is found that the C-7 position, which is far away from the conjugated system, has a strong Cotton effect near 240~260nm and is stronger than the Cotton effect at C-5 position. The Cotton effect of compound 3-5 near 250nm may be a weighted superposition of the Cotton effect of C-7 and C-5 chromophore. The results provide a reference for the study of Gyabgon spiroid sesquiterpene stereotype.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R284;R285
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本文编号：1876645