疣果大戟的化学成分及抗肿瘤活性研究

论文总字数：41441字

摘 要

大戟属是大戟科植物中种类最多、变异性大、分布广泛的一种被子植物，全世界共有2000余种, 其中有80余种分布在我国。大戟植物的化学成分多样复杂，其主要成分为萜类化合物，包括（降碳）倍半萜、二萜、三萜等。此外还包括甾体、花色苷、黄酮、香豆素、鞣质及酸性成分。此外，大戟属植物的主要特征是含有白色或黄白色乳汁, 并具有双重特性:既具有抗菌消炎抗病毒抗结核抗肿瘤以及神经生长因子促进作用等药理活性, 同时表现出对皮肤口腔及胃肠道黏膜强烈的刺激性和致炎、致癌的毒性作用。大戟属植物在中药、民族药理学方面被广泛使用。近年来随着对京大戟、月腺大戟、狼毒大戟、甘遂大戟等大戟属植物的研究愈发深入，新的化合物骨架及其药用价值被不断发现。疣果大戟作为主要分布在我国西北部地区的大戟属草本植物，对其相关研究和报导却比较少。所以我们将疣果大戟作为本文的研究对象。

疣果大戟(Euphorbia micractina Boiss)干燥的根，综合使用了一系列提取、分离方法，包括乙醇提取，有机溶剂萃取，正向硅胶柱层析，薄层色谱法（TLC）等手段从石油醚浸膏中得到了4个纯的化合物单体。然后我们使用现代波谱学方法（¹H-NMR、¹³C-NMR、El-MS、IR）和熔点测定等手段，结合相关文献报导，确定了这4个化合物的结构。

化合物S1是岩大戟内酯A (Jolkinolide A)；化合物S2是β-谷甾醇 (β-sitosterol)；化合物S3是化合物24-亚甲基-环阿屯醇(24-Methylenecycloartanol)；S4是β-谷甾醇葡萄糖苷(β-sitosterol-3-O-β-D-glucoside)；S1属于松香烷型二萜，S2和S4属于甾体，S3属于环阿屯烷型三萜。

用MTT法对化合物S1和S3进行体外抗肿瘤活性试验，结果发现化合物S1和S3对4种肿瘤细胞的IC₅₀值显著高于对照组HCPT，并且大部分值超过了10倍以上。从剂量曲线可以直观地看出S1和S3在与对照组相同剂量下的肿瘤生长抑制率也很低。可以认为化合物S1和S3对这4种肿瘤细胞具有较弱的抗肿瘤活性。深入的化学成分及药理学研究还有待于进一步确认。

疣果大戟是民间重要的药用植物资源，对其化学成分及药理活性的研究有助于药物作用物质基础的了解及活性成分的发现，并为新药开发奠定基础。

关键词：大戟属，疣果大戟，萜类，甾体类，抗肿瘤活性

ABSTRACT

Euphorbia is an angiosperm with the most species, great variability and wide distribution in Euphorbiaceae. There are more than 2000 species in the world, of which more than 80 species are distributed in China. The chemical constituents of Euphorbia officinalis are various and complex. Its main constituents are terpenoids, including sesquiterpenes, diterpenoids and triterpenoids. It also contains steroids, anthocyanins, flavonoids, coumarins, tannins and acidic components. In addition,having white or yellow-white milk as the main characteristics,Euphorbias have dual characteristics: both anti-bacterial, anti-inflammatory, anti-viral, anti-tuberculosis, anti-tumor and nerve growth factor promoting effects, while showing a strong irritation to the skin, oral and gastrointestinal mucosa, inflammation and carcinogenic toxicity. Euphorbia plants are widely used in traditional Chinese medicine and ethnopharmacology. In recent years, with the deepening research on Euphorbias, such as Euphorbia pekinensis, Euphorbia ebracteolata hayata, Euphorbia fischeriana,Euphorbia kansui and so on, new compounds skeleton and their medicinal value have been found continuously. Euphorbia micractina Boiss is mainly distributed in the northwest of China, but there are few studies and reports on it. So we take Euphorbia micractina Boiss as the object of this study.

Dry roots of Euphorbia micractina Boiss, we synthetically used a series of extraction and separation methods, including ethanol extraction, organic solvent extraction,normal phase silica gel column chromatography, thin layer chromatography (TLC) and other means to obtain four pure monomers from petroleum ether extract. Then we used modern spectroscopic methods (¹H-NMR, ¹³C-NMR, El-MS, IR) and melting point determination to determine the structures of these four compounds.

Compound S1 is Jolkinolide A; Compound S2 is beta-sitosterol; Compound S3 is 24-methylene-cycloartanol; Compound S4 is beta-sitosterol-3-O-beta-D-glucoside; Compound S1 belongs to abietane-type diterpenoids; Compound S2 and S4 belong to steroids; Compound S3 belongs to cycloartane-type triterpenoids.

The anti-tumor activity of compounds S1 and S3 in vitro was tested by MTT method. The results showed that the IC₅₀ values of compounds S1 and S3 to four kinds of cancer cells were significantly higher than those of control group HCPT, whom most of the values were 10 times higher than. From the dose curve, it can be seen intuitively that the inhibition rate of S1 and S3 on tumor growth is also very low at the same dose as the control group. It can be concluded that compounds S1 and S3 have weak anti-tumor activity against these four kinds of cancer cells. Further studies on chemical constituents and pharmacology need further confirmation.

Euphorbia officinalis is an important folk medicinal plant resource. The study of its chemical constituents and pharmacological activities will contribute to the understanding of the material basis of drug action and the discovery of active ingredients, and lay a foundation for the development of new drugs.

Key words:Euphorbia,Euphorbia micractina Boiss,terpenoids,steroids,anti-tumor activity

目 录

摘要 I

ABSTRACT II

第一章 绪论 1

1.前言 1

1.1选题背景及意义 1

1.2课题目的 1

1.3课题内容 1

2.大戟属植物化学成分 2

2.1二萜类 2

2.1.1千金烷型 2

2.1.2松香烷型 4

2.1.3阿替斯烷型 6

2.1.4巨大戟烷型 7

2.1.5贝壳杉烷型 8

2.1.6海松烷型 9

2.1.7曼西醇型 10

2.1.8假白榄烷型 13

2.1.9 segetane型 18

2.2三萜类 19

2.2.1甘遂烷型 19

2.2.2大戟烷型 21

2.2.3齐墩果烷型 23

2.2.4乌苏烷型 25

2.2.5羽扇豆烷型 26

2.2.6羊毛脂烷型 28

2.2.7环阿屯烷型 29

2.2.8其他类型 30

3.大戟属植物的药理活性 31

3.1抗肿瘤作用 32

3.2抗菌作用 32

3.3抗炎镇痛作用 33

3.4抗病毒作用 34

3.5小结 35

4. 疣果大戟介绍 35

第二章 疣果大戟根部化合物成分研究 38

1.实验部分 38

1.1实验方法 38

1.1.1溶剂提取法 38

1.1.2有机溶剂萃取 38

1.1.3柱层析 38

1.1.4 薄层色谱法（TLC） 38

1.1.5大孔吸附树脂 38

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