【作者】 张琳；

【导师】 吴立军； 奥山徹；

【作者基本信息】 沈阳药科大学 ， 药物化学， 2010， 博士

【摘要】 本论文分别对板栗总苞的抗糖尿病活性及活性成分以及土耳其植物Seseli hartvigii的化学成分进行了研究。板栗(Castanea mollissima Blume)是壳斗科(Fagacaea)栗属(castanea)的落叶果木,又名栗子、皮栗、风栗、毛栗等,主要分布于华北、西南和长江流域等地。近年来研究表明Castanea属植物具有抗菌、抗氧化、抗肿瘤、延缓衰老、抗血小板凝集等生物活性。课题组前期对板栗的种仁、种皮、板栗花的化学成分进行了全面的化学研究。本论文在前期研究基础上,对板栗种仁、种皮及总苞的总提取物及各萃取层进行了抗糖尿病的活性评价,发现板栗总苞的总提取物及萃取层具有显著的醛糖还原酶(AR)抑制及终末糖化产物(AGEs)生成抑制的活性。因此,本实验在醛糖还原酶抑制活性的指导下,对板栗总苞的活性成分进行了深入研究,利用硅胶柱色谱、反相ODS、制备型HPLC等方法分离得到25个化合物,通过IR、UV、NMR、MS等谱学手段确定了20个化合物的结构,分别鉴定为山柰酚(kaempferol,1)、2,4-二羟基苯甲酸(2,4-dihydroxy-benzoic acid,2)、没食子酸甲酯(methyl-gallate,3)、3,4-二羟基苯甲酸(3,4-dihydroxy-benzoic acid,4)、没食子酸(gallic acid,5)、鞣花酸(ellagic acid,6)、tiliroside (kaempferol-3-O-[6"-O-(E)-p-cou--maroyl]-β-D-glucopyranoside,7)、kaempferol-3-O-[6"-O-(E)-p-coumaroyl]-β-D-galatopy--ranoside (8)、kaempferol-3-O-[2"-O-(E)-p-coumaroyl]-β-D-glucopyranoside (9)、二十九烷醇(1-nonacosanol,10)、豆甾烷-4-烯-3,6-二酮(stigmast-4-en-3,6-dione,11)、没食子酸正丁酯(n-butyl gallate,12)、2,4,6-三羟基苯甲酸(2,4,6-trihydroxy-benzoic acid,13)、kaempferol-3-O-[2",6"-di-O-(E)-p-coumaroyl]-β-D-glucopyranoside (14)、kaempferol-3-O-[2",6"-di-O-(E)-p-coumaroyl]-β-D-galacopyranoside (15)、casuariin (16)、casuarinin(17)、castalagin (18)、苯甲基-6-O-β-D-吡喃葡萄糖苷(benzyl-O-β-D-glucopyranoside,19)、正丁基吡喃木糖苷(n-butyl-O-β-D-xylopyranoside,20),其中化合物2、9、10、12、13、15、19、20为从该属植物中首次分离得到,化合物3、18为从该植物中首次分离得到。活性结果表明各化合物均显示出不同程度的醛糖还原酶抑制活性,其中化合物3、6-9、12、14-20显示出比阳性对照药槲皮素(quercetin)更强的活性,初步阐明了黄酮类及多酚类化合物是抗糖尿病活性的物质基础。另外,对由高血糖引起的多元醇通路激活与终末糖化产物生成而诱发糖尿病并发症的机制进行探讨,并考察ELISA实验中的清洗方式与抗体浓度进行考察,确定最佳方案为采用真空泵吸干式清洗方式,一次抗体与二次抗体浓度均为0.0μg/ml。另外,我们还对土耳其植物Seseli hartvigii进行了化学成分研究。Seseli hartvigii是伞形科,伞形目植物,隶属西风芹属(邪蒿属)。本文对Seseli hartvigii的正己烷提取物与乙酸乙酯提取物进行了化学成分研究,利用硅胶柱色谱、制备型HPLC等方法分离得到12个化合物,通过IR、UV、NMR、MS等谱学手段,结合化学衍生法,确定了11个化合物的结构,分别鉴定为6-(3’-methyl-2’-oxo-3’-butenyl)-7-methoxycoumarin (1)、tamarin (2)、佛手柑素(bergamottin,3)、异欧前胡素(isoimperatorin,4)、disenecionyl cis-khellactone (5)、软木花椒素(suberosin) [6-(3’-methylbutenyl)-7-methoxycoumarin] (6)、川白芷内酯(anomalin,7)、(-)-(3’R,4’R)-3’-angeloyloxy-4’-acetoxy-3’,4’-dihydroseselin (8)、羌活醇(notoptol,9)、[5-(2"E)-6"-hydroxy-3",7"-dimethylocta-2",7"-dienyloxy] psoralen (10)、香柑醇(bergaptol,11),其中化合物1为一新天然产物,化合物2、9-11为从该属植物中首次分离得到,研究结果为该植物的化学研究提供了参考和依据。更多还原

【Abstract】 The anti-diabetic activity and active constituents of involucre of Castanea mollissima Blume, together with constituents from Turkey plants Seseli hartvigii were studied in this paper.Castanea mollissima Blume is a plant belonging to the Castanea genus of Fagacaea family, which was also called chestnuts.It was mainly distributed in North China, Southwest and Changjiang River Valley.According to recent reports,the plants of Castanea genus have the activities of anti-bacteria, antioxidant, anti-cancer, anti-age, antiplatelet-aggregating and so on.The chemical constituents of chestnuts,shells and flowers of it were studied by our group systematically. Base on the previous studies,anti-diabetic activities of all extracts from chestnuts,shells and involucres of C. mollissima Blume were valued by two screening systems in vitro, and extract of involucres showed the significant inhibitory activities on Aldose Reductase (AR) and the formation of Advanced Glycation End Products (AGEs). Under the guidance of AR assay, the active constituents of involucre were studied,25 compounds were obtained by means of silica gel, Sephadex LH-20 column chromatography, ODS column and HPLC isolation methods, and 20 compounds were identified by the IR, UV, NMR, MS spectral analysis. They were elucidated as kaempferol (1), 2,4-dihydroxy-benzoic acid (2),methyl-gallate (3),3,4-dihydroxy-benzoic acid (4), gallic acid (5), ellagic acid (6), tiliroside (kaempferol-3-O-[6"-O-(E)-p-coumaroyl]-β-D-glucopyranoside,7), kaempferol-3-O-[6’-O-(E)-p-coumaroyl]-β-D-galatopyranoside (8), kaempferol-3-O-[2"-O-(E)-p-coumaroyl]-β-D-glucopyranoside (9),1-nonacosanol (10), stigmast-4-en-3,6-dione(11),n-butyl gallate(12),2,4,6-trihydroxy-benzoic acid(13), kaempferol-3-O-[2",6"-di-O-(E)-p-coumaroyl]-β-D-glucopyranoside (14), kaempferol-3-O-[2",6"-di-O-(E):p-coumaroyl]-β-D-galacopyranoside (15), casuariin (16), casuarinin (17), castalagin (18), benzyl-O-β-D-glucopyranoside (19), n-butyl-O-β-D-xylopyranoside (20), respectively. Among themcompounds 2,9,10,12,13,15,19 and 20 were isolated from Castanea genus for the first time, and compound 3 and 18 were obtained from this plant for the first time.The results showed all the compounds exhibited different degrees of AR inhibitory activity, especially compounds 3,6-9,12,14-20 exhibited much stronger activities than that of positive control quercetin, which suggested that flavoniods and polyphenol derivatives were the basis for anti-diabetic activity. The mechanism of complications of diabetes induced by activation of polyhydric alcohols pathway and production of AGEs which caused by hyperglycemia were presumed. In the inhibitory experiments of AGEs assay, the best program was found to use the vaccum pump to wash the wells, and the concentration of the first-antibody and second antibody was 0.05μg/ml.In addition, the chemical study on the plant of Seseli hartvigii from Turkey was carried out by our work. Seseli hartvigii was belonging to Seseli genus of Umbelliferae family.Its constituents of n-hexane and EtOAc soluble fractions were studied.13 compounds were obtained by the silica gel, Sephadex LH-20 column chromatograghy and HPLC isolation methods, among them 12 compounds were identified by IR, UV, NMR and MS spectral analysis, and they were identified as 6-(3’-methyl-2’-oxo-3’-butenyl)-7-methoxycoumarin(1), tamarin (2), bergamottin (3), isoimperatorin (4), disenecionyl cis-khellactone (5), suberosin [6-(3’-methylbutenyl)-7-methoxycoumarin] (6), anomalin (7), (-)-(3’R,4’R)-3’-angeloyloxy-4’-acetoxy-3’,4’-dihydroseselin (8),notoptol(9),5-[(2"E)-6"-hydroxy-3",7"-dimethylocta-2", 7"-dienyloxy] psoralen(10),bergaptol (11),respectively. Among them, compound 1 was a new natural product, compounds 2,9-11 were isolated from Seseli genus for the first time. The results would be helpful for the further study on the chemical studies.更多还原