羽叶鬼灯檠的化学成分及活性研究和两种生物活性筛选模型的应用研究
其他题名Chemical and Active Constituents from Rodgersia pinnata, Investigation of Two Bioassays Used in Natural Products
嵇长久
学位类型硕士
导师谭宁华
2004
学位授予单位中国科学院昆明植物研究所
学位授予地点中国科学院昆明植物研究所
学位专业植物学
关键词羽叶鬼灯檠 虎耳草科 化学成分 活性成分 生物活性筛选模型 抗色素沉积 酪氨酸酶 抗骨质疏松 碳酸配酶ii
摘要本论文涉及一种药用植物——羽叶鬼灯檠(Rodgersia pinnata Franch.)的化学成分及活性研究,和两种体外生物活性筛选模型——抗色素沉积模型TS的建立及其应用和抗骨质疏松模型CA-II在天然产物生物活性筛选方面的应用研究,最后还综述了近年来抗色素沉积药物筛选的方法、抑制剂及其作用机理和酪氨酸酶抑制剂的研究进展等。第一章从羽叶鬼灯檠中共计分离鉴定了27个化合物,其中有4个新化合物,并发现了4个新的活性化合物;第二章建立了以酪氨酸酶为靶标的抗色素沉积筛选模型,并对4775个样品进行了批量筛选,发现了14个活性样品;第三章开展了以CAII为靶标的抗骨质疏松模型的筛选工作,对1393个样品进行了批量筛选,发现了139个活性样品,其中有两个化合物和一个提取物样品活性较强。各章具体内容简介如下:第一章羽叶鬼灯檠的化学成分及活性研究:羽叶鬼灯檠为虎耳草科(Saxifragaceae)鬼灯檠属(Rodgersia Gray)植物,主要分布于中国西南地区云南、四川两省,功效与西南鬼灯檠(R.sambucifolia)相同。前期我们发现它在抗骨质疏松模型CA-IIH及CAT-B和抗肿瘤模型CDC25上显示较强活性,所以对其进行了样品复采和活性追踪实验,发现活性部位主要为大极性的单宁成分,鉴于单宁成分作用的非选择性,于是我们转向对其它成分的分离和活性研究。从R.Pinnata中共计分离鉴定了27个化合物,其中包括6个链状单萜二糖苷类化合物(1-6);2个齐墩果烷型三萜类化合物(7-8);两对苯丙素苷类异构体(9-12);3个岩白菜素类化合物(13-15);其它尚有黄酮类(16)、黄烷类(17-18)、单宁类(19-21)、没食子酸(22)、甾醇类(23-24)及饱和脂肪酸(25-27)等;其中4个是新化合物,包括一个新的单萜二糖苷(E)-3,7-Dimethyl-1-O-[a-L-rhamnopyranosyl-(1→6)-β-D-glucopyranosyll-oct-2-en-7-01(1),一个新的三萜3α-O-(E)-p-Hydroxycinnamoyl,olean-12-en-27-oic acid(7),一对新的苯丙素异构体3,5-Dimethoxy-4-O-β-D-glucopyranosyl-phenylpropane-7,8,9-triol(9,10)。单萜二糖苷、苯丙素苷均为首次在该属植物中发现。化合物1-5,7-8,10,12,13-19,20121,22经过分子筛选模型细胞周期基因25表达的蛋白磷酸酶筛选模型CDC25、组织蛋白酶B筛选模型CAT-B、I型丝氨酸/苏氨酸的蛋白磷酸酶的筛选模型PPI和肿瘤细胞筛选模型人胃腺癌细胞株BGC-823、人肝癌细胞株BEL-7402、人结肠腺癌细胞株HT-29、人前列腺癌细胞株DU-145、人乳腺癌细胞株MCF-7等8个筛选模型的筛选后只发现化合物2、7、13和8具有一定的抗肿瘤活性,2,7、13分别对人胃腺癌细胞株BGC-823的半数抑制浓度(IC50)分别为5.85、9.12、6.97μg/ml,8对人前列腺癌细胞株DU-145的半数抑制浓度(IC50)为8.08μg/ml,这些活性皆为首次报道。第二章抗色素沉积活性筛选模型TS的建立及其批量筛选:酪氨酸酶(Tyrosinase)是黑素生成过程中的关键酶,当前色素异常等疾病严重影响着人们的心理及生理健康,美白产品的开发也是比较火热,所以建立以酪氨酸酶为靶标的抗色素沉积生物活性筛选模型非常重要;有报道植物环肽显示对酪氨酸酶有抑制活性,本实验室在前期研究中从植物中发现了多个植物环肽,为了研究环肽活性,也为了丰富筛选室的筛选模型,故我们建立了酪氨酸酶的筛选模型,并对4775个样品进行了批量筛选,发现了14个活性样品。其中2个活性化合物样品活性较强和1个提取物样品活性较好,即提取物9903058的IC50值为55.2μg/ml,化合物200102123的Icso值为25.9μg/ml和9903155的IC50值为27.5μg/ml。第三章抗骨质疏松活性筛选模型CA-II的批量筛选:碳酸配酶11(CAll)是骨质疏松症的一个重要靶标,所在筛选室已建有其相关筛选模型CA-II,本次共对1393个样品进行了批量筛选,发现了139个活性样品,其中有两个化合物和一个提取物样品活性较强,即化合物2001146的IC5o=0.30μg/ml,200111132的IC50=0.37μg/ml,提取物2001034的IC50=0.74μg/ml。 第四章抗色素沉积药物活性筛选的研究进展:本章综述了近年来抗色素沉.积药物筛选方法、抑制剂及其作用机理和酪氨酸酶抑制剂的研究进展等。
其他摘要The dissertation includes three parts. The first part is about the chemical and active chemical constituents from Rodgersia pinnata Franch. (Saxifragaceae), from which 27 compounds, including 4 new compounds and 4 new active compounds, were isolated and identified. The second is about the establishment of depigment bioassay TS, together with the application of TS and anti-osteoporosis bioassay CA-II used in natural products. In TS assay, 4775 samples were tested and 14 samples were found with inhibitory activity on tyrosinase. In CA-II assay, 1393 samples were tested with 139 samples showed inhibitory activity on Carbonic anhydrase II. The third is about the progress of depigment drugs R & D including melanogenesis pathway, methods in screening depigment drugs, class of depigmentation and tyrosinase inhibitors as well as their mechanism. Chapter 1 Chemical and Active Constituents from Rodgersia pinnata Rodgersia pinnata Franch. (Saxifragaceae) is mainly distributed in Yunnan and Sichuang Province, Southwest of China, which has the same efficacy as R. sambucifolia used as folk herbs. We found the extracts and fractions of the plant with strong inhibitory activities on the anti-osteoporosis model of CAT-B Assay, CA-II Assay and anti-tumor model of CDC25 Assay in previous works. So its recollection and bioassay-guided separation was conducted, unfortunately it was discovered that the tannin-like fractions were the main active constituents. We turned attentions to other active constituents for the non-selective activity of tannins on many enzymes or proteins. Finally 27 compounds were isolated and identified, which belong to acyclic monoterpene disaccharide glycosides (1-6), oleane triterpenoids (7-8), phenylpropane glycosides (9-12), bergenins (13-15), flavanoids (16-18), tannins (19-21) and others (22-27). Among them, (E)-3,7-DimethyI-l-0-[α-L-rhamnopyranosyI-(l→6)-β-D-glucopyranosyl]-oct-2-en-7-ol (1), 3α-0-(E)-β-Hydroxycinnamoyl-olean-12-en-27-oic acid (7) and 3,5-Dimethoxy-4-0-β-D-glucopyranosyl-phenylpropane-7,8,9-triol (9, 10) are new compounds. It's the first time that monoterpene disaccharide glycosides and phenylpropane glycosides were found in the genus. Compounds 1-5, 7-8, 10, 12, 13-19, 20/21, 22 were tested on the bioassays of CDC25, CAT-B, PP1 (metabolism disease), and cancer cell line test on BGC-823, BEL-7402, HT-29, DU-145, MCF-7 cell lines. Finally Compounds 2, 7, 13 showed anti-tumor activities on BGC-823 with IC50 at 5.85, 9.12 and 6.97μg/ml, respectively, and Compound 8 showed anti-tumor activity on DU-145 with IC50 at 8.08ug/ml, which are also reported for the first time. Chapter 2 Establishment and Application of Depigment Bioassay TS in the Screening of Natural Products Tyrosinase is one of key enzymes in melanogenesis pathway and also one of main targets in searching for depigment drugs. Based on literatures, we established the tyrosinase assay (TS). With this assay 3284 extracts or fractions and 1491 compounds were tested. Finally 8 extracts or fractions and 6 compounds showed inhibitory activities in the concentrations of 250μg/ml and 62.5μg/ml, respectively. Chapter 3 Application of Anti-osteoporosis Bioassay CA-II in the Screening of Natural Products. Carbonic anhydrase II is an important enzyme target in osteoporosis. Assay on carbonic anhydrase II was set up in our screening laboratory a few years ago. Herein 1086 extracts or fractions and 307 compounds were tested. Finally 125 extracts or fractions and 14 compounds showed inhibitory activities in the concentrations of 10μg/ml and 12.5μg/ml, respectively, in which two compounds 2001146 and 200111132 showed significant activities with IC50 at 0.30 and 0.37μg/ml, and extract 2001034 IC50 at 0.74μg/ml, respectively. Chapter 4 Progress in the Screening of Depigment Drugs Pigmentation is an important issue nowadays. In this chapter the progress of depigment drugs R & D including melanogenesis pathway, methods in screening depigment drugs, class of depigment and tyrosinase inhibitors as well as their inhibitory mechanism were reviewed.
页数77
语种中文
文献类型学位论文
条目标识符http://ir.kib.ac.cn/handle/151853/728
专题昆明植物所硕博研究生毕业学位论文
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嵇长久. 羽叶鬼灯檠的化学成分及活性研究和两种生物活性筛选模型的应用研究[D]. 中国科学院昆明植物研究所. 中国科学院昆明植物研究所,2004.
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