【作者】 马洁；

【导师】 苏亚伦；

【作者基本信息】 北京协和医学院 ， 药物化学， 2012， 博士

【摘要】 新疆产贯叶金丝桃Hypericum perforatum L和腺点金丝桃Hypericum scabrum L.为藤黄科金丝桃属药用植物。金丝桃属植物因其具有独特的化学成分和多样的药理活性,所以一直是现代药物研究的热点。为了从民间传统中草药的近源植物资源中寻找类型多样、结构新颖的代谢产物,为药理活性筛选提供丰富多样的植物代谢产物样品,通过随即筛选寻找强活性化合物,为化学合成和创新药物提供有价值的模板结构,同时,也为揭示传统和民间中草药的成分组成特点和和近源植物资源的利用提供科学基础,本学位论文分别对贯叶金丝桃和腺点金丝桃乙醇提取物的化学成分进行了系统的分离和结构鉴定,并通过与药理学人员的合作,对分离过程中得到的不同部位以及单体化合物的活性进行了初步药理活性筛选。运用硅胶、大孔树脂、凝胶Sephadex LH-20、中压液相色谱及高效制备液相色谱等各种色谱分离手段,从贯叶金丝桃Hypericum perforatum L的乙醇提取物中,分离得到32个化合物,通过MS、UV、IR、1D NMR、2D NMR等波谱分析及化学方法,鉴定了它们的结构,发现新化合物2个(1*,2*),1‘为新的酚酸类化合物,化合物2‘为新的黄酮类化合物。这些化合物包括黄酮类化合物10个(2‘,14-18,20-21,27-28),口山酮类化合物2个(12,19),酚酸及其苷类化合物7个(1‘,5-7,10,25-26),萜类化合物5个(3-4,22-24),其他类型化合物4个(31-32,13,29)。化合物3,4,7,31均为首次从金丝桃属植物中分离得到,化合物5,6为首次从贯叶金丝桃植物中分离得到。在抗糖尿病(对蛋白质酪氨酸磷酸酶1B的抑制活性)的筛选中,化合物8在2μmol·L-1浓度时,对基因重组人PTP1B的抑制率为96.4%,IC50为2.5μM。5个化合物(1,3,4,9,16)可能具有较高的NE再摄取抑制活性。针对金丝桃属植物中含有的特征性成分间苯三酚类化合物,同时对近源植物资源的利用的研究,我们另选择了我国特有植物新疆产腺点金丝桃进行研究,采用多种色谱学手段和波谱学技术从该植物乙醇提取物中的石油醚萃取部位分离39个化合物,鉴定了38个(1-38),发现新化合物20个(1*-20*),其中新化合物19个,新天然产物1个,间苯三酚类化合物34个,倍半萜类化合物2个,其它类3个。其中新化合物包括首次发现的降碳的含有缩酮单元的间苯三酚类化合物10*-13*,降碳的含有螺原子螺[4,5]的间苯三酚类化合物14*内酯型间苯三酚类化合物8*,9*,还有单一间苯三酚类化合物1*-6*,呋哺型衍生物15*-17*,吡喃型衍生物7‘,含有氧桥的衍生物19*,化合物18**为新天然产物,另外还有一个倍半萜新化合物20*。运用CD光谱对间苯三酚类化合物的立体构型进行了初步探讨和确定。利用特征CD数据(尤其是p,丫-不饱和片段所产生的CD数据(300nm附近)),对含有类似结构单元化合物的构型确定提供了一种相对简便的方法。在多种药理模型上对分离鉴定的部分单体化合物进行了活性筛选,在大鼠脑突触体[3H]-5-HT再摄取活性筛选实验中,有化合物33(34)的抑制率大于50%,提示上述化合物可能具有较高的5-HT再摄取抑制活性。在大鼠脑突触体[3H]-NE再摄取活性筛选实验中,腺点金丝桃各部位均具有显著的NE再摄取抑制活性。18个化合物(3*,5*,8*,9*,10*,11*,14*,18*,23,25,26,27,28,30,32,33(34),36,37(38)的抑制率大于80%,提示可能具有较高的NE再摄取抑制活性。对肝细胞损伤的保护作用筛选中,结果显示化合物2*,5*,7*,8*,23,25,29,32,37(38)在10,μM浓度水平均对肝细胞有保护作用,成活率分别为67%,77%,78%,73%,84%,94%,69%,80%,81%,与模型组比较有显著性差异。细胞毒活性筛选结果显示腺点金丝桃的水部位对HCT-8细胞有一定的细胞毒作用,石油醚萃取部位对Bel-7402、BGC-823和A2780有一定的细胞毒作用。但是随后的单体化合物筛选结果显示仅33(34)对肿瘤细胞有一定的细胞毒作用,其它均未显示有细胞毒活性。在抗糖尿病(对蛋白质酪氨酸磷酸酶1B的抑制活性)高通量模型的筛选中,分离得到的化合物在10μM浓度下,均未显示明显的活性。更多还原

【Abstract】 Hypericum perforatum L. and Hypericum scabrum L. are both species of the genus Hypericum of the Guttiferae family, which were collected from Xinjiang Uygur Autonomous Region. Because of their unique chemical compositions and a variety of pharmacological activity, study on this Hypericum has been a hot spot for the modern drug research in the pharmacological and phytochemical field.As part of a program to assess the chemical and biological diversity of several traditional Chinese medicines, this dissertation investigated chemical constituents of the aerial parts of Hypericum perforatum L. and Hypericum scabrum L., and bioactivities of different extracts and purified compounds on several pharmacological models with the help of pharmacological staff.By using various chromatographic techniques,32compounds have been isolated from an ethanolic extract of the aerial parts of Hypericum perforatum L. and their structures were elucidated by spectroscopic methods. Among them, two compounds were new (1,2*), including one phenolic compound (1*) and a flavonoid compound (2*). These compounds were divided into ten flavonoids (2,14-18,20-21,27-28), two xanthones (12,19), seven phenolic acids and their glycosides (1*,5-7,10,25-26), five terpenoids (3-4,22-24) and other types of compounds (31-32,13,29). Compound3,4,7,31were obtained from Hypericum for the first time and compound5,6was isolated from this plant for the first time. At a concentration of2μmol·L-1, compound8inhibited recombinant human PTP1B with inhibitory rate of96.4%and IC50of2.5μM. Five compounds (1*,3,4,9,16) may have NE reuptake inhibitory activity.In pursuit of another type of characteristic phloroglucinol components, which were not found in the research on H. perforatum, our study group have found39compounds from Petroleum ether extraction of aerial parts of H. scabrum, and identified38compounds, which included34polyprenylated phloroglucinols, two sesquiterpenes and two other kinds of compounds. Among them, compounds1-17*,19*-20*were new products,18*were new natural product, compounds1-6were simple phloroglucinols,8*and9were novel tricyclo [5.2.1] phloroglucinol compounds,10*and13*were unusual decarbon phloroglucinol components containing the ketal unit,14*was an unusual phloroglucinol component containing the spiro [4,5] unit,15*and17*were furan-phloroglucinols,7*was a pyran-phloroglucinol derivative,19*was a phloroglucinol containing oxygen bridge respectively, as well as a new sesquiterpene compound20*. A preliminary study to determine the stereochemistry of the CD spectra of phloroglucinol components was proposed. The feature of CD data [especially (3, y-unsaturated fragment of the CD data (near300nm)] provided a relatively easy way to determine the configuration for those compounds with similar structural units.In the in vitro assays, compound33(34) showed obvious [3H]-5-HT reuptake inhibitory activity. In the rat brain synaptosomes [3H]-NE uptake activity screening experiments, all kinds of crude extracts of H. scabrum exhibited significant NE reuptake inhibitory activity. The inhibition rates of18compounds (3*.9*,10*,11*,14*18*,23,25,26,27,28,30,32,33(34),36,37(38)) were all higher than80%. suggesting that these compounds inhibited remarkable NE reuptake inhibitory activity. In the10μM concentration compounds2*,5*,7*,8*,23,25,29,32,37(38) exhibited protective activity against a damage of hepatocyte, the survival rates were67%.77%,78%,73%,84%,94%,69%,80%,81%, with significance compared to the blank control group. The water part and the petroleum ether extraction showed potent cytotoxicity. However, purified compounds did not show significant cytotoxicity activity and PTP1B inhibition activity.更多还原