【作者】 史琳；

【导师】 赵余庆；

【作者基本信息】 沈阳药科大学 ， 天然药物化学， 2010， 博士

【摘要】 绞股蓝为葫芦科(Cucurbitaceae)绞股蓝属植物kg绞股蓝[Gynostemma pentaphyllum (Thunb.) Makino]的干燥全草,为多年生攀缘草本,主要分布于中国、日本、朝鲜等国。绞股蓝是五加科外具有人参皂苷资源的植物之一,具有滋补保健、抗癌防衰、增强体质和改善脂质代谢等多种功能,被誉为“南方人参”。本文在综述了绞股蓝的化学成分和药理活性研究的基础上,对其进行了系统的化学成分研究及抗肿瘤活性研究。采用反复硅胶柱色谱法、大孔树脂、Sephadex LH-20、反相ODS、半制备型HPLC等多种色谱手段从绞股蓝的75%乙醇提取物中,分离得到38个化合物。并通过理化性质、波谱数据分析(IR、NMR、MS等)和化学方法鉴定了它们的结构,分别为：(23S)-21R-O-正丁基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-比喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(1),(23S)-21S-O-正丁基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(2),(23S)-21R-O-正丁基-19-醛基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(3),(23S)-21S-O-正丁基-19-醛基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(4),(23S)-21R-O-正丁基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(5),(20S)-3β,20,21-三羟基达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(6),长梗绞股蓝皂苷Ⅰ(7),α-菠菜甾醇(8),β-谷甾醇(9),α-菠菜甾醇3-O-β-D-吡喃葡萄糖苷(10),胡萝卜苷(11),绞股蓝皂苷Ⅲ(人参皂苷Rb1,12),绞股蓝皂苷Ⅳ(人参皂苷Rb3,13),绞股蓝皂苷Ⅷ(人参皂苷Rd,14),绞股蓝皂苷ⅩⅣ(15),绞股蓝皂苷Ⅻ(人参皂苷F2,16),绞股蓝皂苷ⅩⅢ(17),芸香苷(18),丙二酸(19),(20R,23R)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-6-O-乙基-β-D-吡喃葡萄糖苷(20),(20S,23S)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-6-O-乙基-β-D-吡喃葡萄糖苷(21),绞股蓝皂苷XLIX (22),(20S)-3β,20,21-三羟基达玛烷-24-烯3-O-{[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖基}-21-O-β-D-吡喃葡萄糖苷(23),(23S,23S)-19-醛基-3β,20ξ,21,26-四羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(24),(23R,23R)-19-醛基-3β,20ξ,21,26-四羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(25),(20R,23R)-19-醛基-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(26),(20S,23S)-19-醛基-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(27),(20S)-3β,20,21-三羟基达玛-23,25-二烯3-O-{[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖基}-21-O-β-D-吡喃葡萄糖苷(28),(20S)-19-醛基-3β,20,21-三羟基达玛-23,25-二烯3-O-{[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖基}-21-O-β-D-吡喃葡萄糖苷(29),(23S)-21R-O-乙基-19-醛基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(30),(23S)-19-醛基-3β,20ξ,21ξ-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(31),(20S)-19-醛基-3β,20,21ξ,25-四羟基-21,24ξ-环达玛烷3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(32),(20R)-19-醛基-3β,20,21ξ,23ξ-四羟基-21,24ξ-环达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(33),(20R,23R)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(34),(20S,23S)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(35),(20S,23S)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(36), (20R,23R)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[β-D-吡喃木糖基(1→3)][β-D-吡喃葡萄糖苷(37),槲皮素(38)。其中未见文献报道的新化合物14个,分别为化合物1-6、24-29、36和37。采用MTT法,对从绞股蓝中得到的部分化合物的体外肿瘤抑制活性进行了初步筛选和评价。化合物1-5,26,33,36-37对细胞株HL-60均具有不同程度的细胞生长抑制作用,其中化合物3活性最强,IC50值为8.45μmol/L.化合物1-5,7对细胞株Colon205和Du145不同浓度下均具有生长抑制作用,其中化合物2对Colon 205的活性最强,IC50值为22.95μmol/L,化合物3对Du145的活性最强,IC50值为17.41μmol/L。化合物20-21,34-35对细胞株A2780和OVCAR-3在不同浓度下均具有生长抑制作用,其中化合物20和34对A2780有较强的活性,其IC50均为10.4μmol/L,化合物20对OVCAR-3有较强的活性,其IC50为2.8μmol/L。更多还原

【Abstract】 Gynostemma pentaphyllum (Thunb.) Makino (Cucurbitaceae) is a herbal medicine with anticancer activity, widely distributed in China, Korea and Japan. It is famous as "Southen Ginseng". The biological active constituents are dammarane-type glycosides, called gypenosides, which are structurally correlated to the ginseng saponins. Pharmacological investigation demonstrated that extracts of this plant exhibited a variety of biological effects, such as anticancer, anti-inflammatory and cadiovascular effects. In our serious of studies on the anticancer natural medicines, we have found some active compounds. As a continuation of our work for discovering more effective components, we have now investigated G. pentaphyllum (Thunb.) Makino, which is the first example of ginsenosides (Rb1, Rb3, Rd, etc.) ever found from a plant not belonging to the Araliaceae. Based on the review on the chemical constituents and pharmacological research of G. pentaphyllum, the chemical constituents were studied and their antitumor activities were tested.By means of many different chromatographic methods such as silica gel, Sephadex LH-20, reversed phase ODS column and P-HPLC chromatography, 38 compounds were isolated from the 75% ethanol extract of the herbs of Gynostemma pentaphyllum. On the basis of spectroscopic analysis (UV, IR, NMR, MS) and physicochemical characters, they were identified as (23S)-21R-O-n-butyl-3β,20ξ,21-trihydroxy-21,23-epoxydammar-24-ene 3-O-[a-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (1), (23S)-21S-O-n-butyl-3β,20ξ,21 -trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl-(1→2)][β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (2), (23S)-21R-O-n-butyl-19-oxo-3β,20ξ,21 -trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rharnnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (3), (23S)-21S-O-n-butyl-19-oxo-3β,20ξ,21 -trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyran-osyl(1→3)]-α-L-arabinopyranoside (4), (23S-21R-O-n-butyl-3β,20ξ,21 -trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (5), (20S)-3β,20,21-trihydroxydammar-24-ene 3-O-[α-L-rhamnopyranosyl-(1→2)][β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (6), gylongiposideⅠ(7),α-spinaster-ol (8),β-sitosterol (9),α-spinasterol 3-Oβ-β-D-glucopyranoside (10), daucosterol (11), gypenosideⅢ(ginsenoside Rb1, 12), gypenosideⅣ(ginsenoside Rb3, 13), gypenosideⅧ(ginsenoside Rd, 14), gypenosideⅪⅤ(15), gypenosideⅫ(ginsenoside F2,16), gypenosideⅩⅢ(17), rutin (18), malonic acid (19), (20R,23R)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-0-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-6-O- acetyl-β-D-glucopyranoside (20), (20S,23S)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-6-O-acetyl-β-D-glucopyranoside (21), gypenoside XLIX (22), (20S)-3β,20,21-trihydroxydarnmar-24-ene 3-O- {[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-β-D-glucopyranosyl}-21 -O-β-D-glucopyranoside (23), (23S,23S)-19-oxo-3β,20ξ,21,26-tetrahydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (24), (23R,23R)-19-oxo-3β,20ξ,21,26-tetrahydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (25), (20R,23R)-19-oxo-3β,20-dihydroxydammar-24-en-21-oci acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl-(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (26), (20S,23S)-19-oxo-3β,20-dihydroxydammar-24-en-21-oci acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (27), (205)-3β,20,21-trihydroxydam-mar-23,25-diene 3-0-{[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)] -β-D-glucopyranosyl}-21-O-β-D)-glucopyranoside (28), (205)-19-oxo-3β,20,21 -trihydroxydam-mar-23,25-diene 3-O-{[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-a-L-arabinopyranosyl}-21-O-β-D-glucopyranoside (29), (23S)-21R-O-ethyl-19-oxo-3β,20ξ,21-trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyran-osyl(1→3)]-α-L-arabinopyranoside (30), (235)-19-oxo-3β,20ξ,21ξ-trihydroxy-21,23-epoxyda-mmar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(13)]-α-L-arabino-pyranoside (31), (20S)-19-oxo-3β,20,21ξ,25-tetrahydroxy-21,24ξ-cyclodammarane 3-0-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (32), (20R)-19-oxo-3β,20,21ξ,23ξ-tetrahydroxy-21,24ξ-cyclodammar-24-ene 3-O-[α-L-rham-nopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (33), (20R,23R)-3β, 20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (34), (205,235)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl-(1→3)]-β-D-glucopyranoside (35), (205,235)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (36), (20R,23R)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (37), quercetin (38). Among them, compounds 1-6, 24-29, 36 and 37 were determined as new compounds by chemical and spectroscopic methods.In the course of our search for triterpene saponins with antitumor activity, by MTT methods, the in vitro cytotoxicity activities against cancer cell lines of some compounds isolated were assayed. The effects of compounds on human cancer cells growth, expressed as the IC50 value, for HL-60 cells lines compounds 1-5,26,33,36-37 showed growth inhibitory effects and differentiation induction abilities, compound 3 was the most potent with the IC50 value of 8.45μmol/L. For Colon 205 and Du 145 cell lines 1-5, and 7 showed growth inhibitory effects and differentiation induction abilities, compound 2 showed strong against Colon 205 cell lines with IC50 value of 22.95μmol/L, and compound 3 showed strong against Du 145 cell lines with IC50 value of 17.41μmol/L. For A2780 and OVCAR-3 cells lines compounds 20,21,34 and 35 showed growth inhibitory effects and differentiation induction abilities, compound 20 showed moderate antitumor activities against A2780 and OVCAR-3 cell lines with IC50 value of 10.40μmol/L and 2.8μmol/L, respectively.更多还原