【作者】 杨凤英；

【导师】 沈家祥； 苏艳芳；

【作者基本信息】 天津大学 ， 应用化学， 2010， 博士

【摘要】 小桃儿七是毛莨科嚏根草属植物铁筷子(Helleborus thibetanus)的干燥根及根茎,性凉味苦,有小毒,具有清热解毒、消肿止痛、活血散瘀之功效,主治尿道炎、膀胱炎、跌打损伤及疮疖肿毒等症。小桃儿七为陕西七药的一种,国内外关于该药材的化学成分及生物活性少有报道,其化学成分及活性成分尚不明确,因此我们对该药材进行了较为系统的化学成分与生物活性的研究。运用溶剂提取和萃取、各种柱色谱法(包括正相硅胶、反相硅胶、聚酰胺、葡聚糖凝胶、大孔吸附树脂等)、高效液相色谱法、溶剂处理及重结晶等方法,从该药材的乙醇提取物中共分离得到三十一个单体化合物,酶水解得到两个苷元,综合运用核磁共振波谱、质谱、红外光谱、紫外光谱、元素分析、水解(包括酶水解和化学水解)以及与标准品的TLC对照等方法鉴定了其中二十八个化合物的结构,分别为:谷甾醇(HTP-C1)、胡萝卜苷(HTE-C1)、deglucohellebrin(HTE-C2)、polypodine B(HTE-C3)、20-羟基蜕皮甾酮(HTE-C4)、14β,16β-二羟基-3β-[(β-D-葡萄吡喃糖基)氧]-5α-蟾酥甾-20,22-二烯(HTE-C5)、14β-羟基-3β-[β-D-葡萄吡喃糖基-(1→6)-(β-D-葡萄吡喃糖基)氧]-5α-蟾酥甾-20,22-二烯(HTB50-C1)、孕甾-5,16-二烯-20-酮-3β-基氧-α-L-鼠李吡喃糖基-(1→2)-氧-[β-D-葡萄吡喃糖基-(1→4)]-β-D-半乳吡喃糖苷(HTB50-C2)、(23S,24S)-24-[(氧-β-D-呋吡喃糖基)氧]-3β,23-二羟基螺甾-5,25(27)-二烯-1β-基氧-(4-氧-乙酰基-α-L-鼠李吡喃糖基)-(1→2)-氧-[β-D-木吡喃糖基-(1→3)]-α-L-阿拉伯吡喃糖苷(HTB50-C3)、(25R)-26-[(氧-β-D-葡萄吡喃糖基)氧]-呋甾-5,20(22)-二烯-3β-基氧-α-L-鼠李吡喃糖基-(1→2)-氧-[β-D-葡萄吡喃糖基-(1→4)]-β-D-半乳吡喃糖苷(HTB50-C5)、5β,14β-二羟基-19-醛基-3β-[(α-L-葡萄吡喃糖基)氧]蟾酥甾-20,22-二烯(HTB30-C1)、5β,14β,16β-三羟基-19-醛基-3β-[(α-L-鼠李吡喃糖基)氧]蟾酥甾-20,22-二烯(HTB30-C2)、hellebrin(HTB30-C3)、(25S)-22α,25-环氧-26-[(氧-β-D-葡萄吡喃糖基)氧]-3β-羟基呋甾-5-烯-1β-基氧-α-L-阿拉伯吡喃糖苷(HTB30-C4)、26-氧-β-D-葡萄吡喃糖基-3β-羟基-22α-甲氧基呋甾-5,25(27)-二烯-1β-基磺酸盐(HTB30-C5)、腺苷(HTB30-C6)、3β-羟基螺甾-5,25(27)-二烯-1β-基磺酸盐(HTB30-C7)、3β-羟基孕甾-5,16-二烯-20-酮-1β-基磺酸盐(HTB30-C8)、5β,14β,16β-三羟基-19-醛基-3β-[(β-D-葡萄吡喃糖基)氧]蟾酥甾-20,22-二烯(HTB30-C9)、14β,16β-二羟基-3β-[β-D-葡萄吡喃糖基-(1→4)-(β-D-葡萄吡喃糖基)氧]-5α-蟾酥甾-20,22-二烯(HTB30-C11)、(23S,24S)-24-{[氧-β-D-葡萄吡喃糖基-(1→4)-β-D-呋吡喃糖基]氧}-3β,23-二羟基螺甾-5,25(27)-二烯-1β-基氧-(4-氧-乙酰基-α-L-鼠李吡喃糖基)-(1→2)-氧-[β-D-木吡喃糖基-(1→3)]-α-L-阿拉伯吡喃糖苷(HTB30-C12a)、(23S,24S)-24-{[氧-β-D-葡萄吡喃糖基-(1→4)-β-D-呋吡喃糖基]氧}-3β,23-二羟基螺甾-5,25(27)-二烯-1β-基氧-β-D-芹呋喃糖基-(1→3)-氧-(4-氧-乙酰基-α-L-鼠李吡喃糖基)-(1→2)-α-L-阿拉伯吡喃糖苷(HTB30-C12b)、(23S,24S)-24-{[氧-β-D-葡萄吡喃糖基-(1→4)-β-D-呋吡喃糖基]氧}-3β,23-二羟基螺甾-5,25(27)-二烯-1β-基氧-β-D-芹呋喃糖基-(1→3)-氧-(4-氧-乙酰基-α-L-鼠李吡喃糖基)-(1→2)-氧-[β-D-木吡喃糖基-(1→3)]-α-L-阿拉伯吡喃糖苷(HTB30-C13a)、(23S)-26-氧-β-D-葡萄吡喃糖基-3β,23-二羟基呋甾-5,20(22),25(27)-三烯-1β-基磺酸盐(HTB30-C14)、葡萄糖(HTB30-C15)、(23S,24S)-24-[(氧-β-D-葡萄吡喃糖基)氧]-3β,23-二羟基螺甾-5,25(27)-二烯-1β-基氧-(4-氧-乙酰基-α-L-鼠李吡喃糖基)-(1→2)-氧-[β-D-木吡喃糖基-(1→3)]-α-L-阿拉伯吡喃糖苷(HTB30-C17)、(23S,24S)-21-羟甲基-24-{[氧-β-D-葡萄吡喃糖基-(1→4)-β-D-呋吡喃糖基]氧}-3β,23-二羟基螺甾-5,25(27)-二烯-1β-基氧-β-D-芹呋喃糖基-(1→3)-氧-(4-氧-乙酰基-α-L-鼠李吡喃糖基)-(1→2)-氧-[β-D-木吡喃糖基-(1→3)]-α-L-阿拉伯吡喃糖苷(HTB30-C19)、3β,14β,16β-三羟基-5α-蟾酥甾-20,22-二烯( HTE-C5-S1 )、3β,14β-二羟基-5α-蟾酥甾-20,22-二烯(HTB50-C1-S1)。上述化合物中有新化合物十四个,包括四个5α-蟾蜍二烯羟酸内酯类化合物(HTE-C5、HTE-C5-S1、HTB50-C1和HTB30-C11)、两个孕甾烷类化合物(HTB50-C2和HTB30-C8)、六个螺甾烷醇类化合物(HTB30-C7、HTB30-C12a、HTB30-C12b、HTB30-C13a、HTB30-C17和HTB30-C19)和两个呋甾烷醇类化合物(HTB30-C5和HTB30-C14)。我们对分离得到的部分化合物进行了牛肾上腺髓质细胞儿茶酚胺释放影响的活性筛选,结果显示,HTE-C2和HTE-C5促进儿茶酚胺释放,具有交感神经兴奋活性。HTE-C3和HTE-C4对儿茶酚胺分泌均无明显作用。本文还对近年来国内外对毛莨科嚏根草属植物化学成分和生物活性的研究进展进行了综述。更多还原

【Abstract】 The roots and rhizomes of Helleborus thibetanus Franch., locally known as‘XiaoTaoErQi’, is endemic to China and used for treatment of cystitis, urethritis, sores and traumatic injury in Chinese folk medicine. So far, there have been few reports on the chemical studies of H. thibetanus, and the chemical and active principles are not clear. Therefore, we have conducted studies on the chemical constituents of the title plant.Thirty-one compounds were isolated from the ethanol extract of the roots of H. thibetanus employing solvent extraction, various column chromatography (including silica gel, polyamide, Sephadex LH-20 and D101 macroporous resin), preparative high performance liquid chromatography (pre-HPLC), solvent washing and recrystallization. Two aglycones were obtained through enzymatic hydrolysis. On the basis of chemical and spectral methods including NMR, MS, IR, UV, ICP-MS, hydrolysis, and comparison with authentic samples, the structures of twenty-eight compounds were identified as sitosterol (HTP-C1), daucosterol (HTE-C1), deglucohellebrin (HTE-C2), polypodine B (HTE-C3), 20-hydroxyecdysone (HTE-C4), 14β,16β-dihydroxy-3β-[(β-D-glucopyranosyl)oxy]-5α-bufa-20,22-dienolide (HTE-C5), 14β-hydroxy-3β-[β-D-glucopyranosyl-(1→6)-(β-D-glucopyranosyl)oxy]-5α-bufa-20,22-dienolide (HTB50-C1), pregna-5,16-diene-20-one-3β-yl O-α-L-rhamnopyranosyl- (1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-galactopyranoside(HTB50-C2), (23S,24S) -24-[(O-β-D-fucopyranosyl)oxy]-3β,23-dihydroxyspirosta-5,25(27)-diene-1β-yl O-(4- O-acetyl-α-L-rhamnopyranosyl)-(1→2)-O-[β-D-xylopyranosyl-(1→3)]-α-L-arabinopy-ranoside (HTB50-C3), (25R)-26-[(O-β-D-glucopyranosyl)oxy]-furosta-5,20(22)- diene-3β-yl O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D- galactopyranoside (HTB50-C5), 5β,14β-dihydroxy-19-oxo-3β-[β-D-glucopyranosyl)- oxy]bufa-20,22-dienolide (HTB30-C1), 5β,14β,16β-trihydroxy-19-oxo-3β-[(β-D- rhamnopyranosyl)oxy]bufa-20,22-dienolide (HTB30-C2), hellebrin (HTB30-C3), (25S)-22α,25-expoxy-26-[(O-β-D-glucopyranosyl)oxy]-3β-hydroxyfurosta-5-ene-1β- yl O-α-L-arabinopyranoside (HTB30-C4), 26-O-β-D-glucopyranosyl-3β-hydroxy-22α- methoxyfurosta-5,25(27)-diene-1β-yl sulfonate (HTB30-C5), adenoside (HTB30-C6), 3β-hydroxyspirost-5,25(27)-diene-1β-yl sulfonate (HTB30-C7), 3β-hydroxypregna- 5,16-diene-20-one-1β-yl sulfonate (HTB30-C8), 5β,14β,16β-trihydroxy-19-oxo-3β- [(β-D-glucopyranosyl)oxy]bufa-20,22-dienolide (HTB30-C9), 14β,16β-dihydroxy-3β- [β-D-glucopyranosyl-(1→4)-(β-D-glucopyranosyl)oxy]-5α-bufa-20,22-dienolide (HTB30-C11), (23S,24S)-24-{[O-β-D-glucopyranosyl-(1→4)-β-D-fucopyranosyl]oxy} -3β,23-dihydroxyspirosta-5,25(27)-diene-1β-yl O-(4-O-acetyl-α-L-rhamnopyranosyl)- (1→2)-O-[β-D-xylopyranosyl-(1→3)]-α-L-arabinopyranoside (HTB30-C12a), (23S, 24S)-24-{[O-β-D-glucopyranosyl-(1→4)-β-D-fucopyranosyl]oxy}-3β,23-dihydroxy- spirosta-5,25(27)-diene-1β-yl O-β-D-apiofuranosyl-(1→3)-O-(4-O-acetyl-α-L-rhamn- opyranosyl)-(1→2)-O-α-L-arabinopyranoside (HTB30-C12b), (23S,24S)-24-{[O-β-D- glucopyranosyl-(1→4)-β-D-fucopyranosyl]oxy}-3β,23-dihydroxyspirosta-5,25(27)-di-ene-1β-yl O-β-D-apiofuranosyl-(1→3)-O-(4-O-acetyl-α-L-rhamnopyranosyl)-(1→2)- O-[β-D-xylopyranosyl-(1→3)]-α-L-arabinopyranoside (HTB30-C13a), (23S)-26-O-β-D-glucopyranosyl-3β,23-dihydroxyfurosta-5,20(22),25(27)-triene-1β-yl sulfonate (HTB30-C14), glucose (HTB30-C15), (23S,24S)-24-[(O-β-D-glucopyranosyl)oxy]- 3β,23-dihydroxyspirosta-5,25(27)-diene-1β-yl O-(4-O-acetyl-α-L-rhamnopyranosyl)- (1→2)-O-[β-D-xylopyranosyl-(1→3)]-α-L-arabinopyranoside (HTB30-C17), (23S,24S) -21-hydroxymethyl-24-{[O-β-D-glucopyranosyl-(1→4)-β-D-fucopyranosyl]oxy}-3β, 23-dihydroxyspirosta-5,25(27)-diene-1β-yl O-β-D-apiofuranosyl-(1→3)-O-(4-O- acetyl-α-L-rhamnopyranosyl)-(1→2)-O-[β-D-xylopyranosyl-(1→3)]-α-L-arabinopyra-noside (HTB30-C19), 3β,14β,16β-trihydroxy-5α-bufa-20,22-dienolide (HTE-C5-S1) and 3β,14β-dihydroxy- 5α-bufa-20,22-dienolide (HTB50-C1-S1).Fourteen new compounds were obtained from the title plant. Among these new compounds, HTE-C5, HTE-C5-S1, HTB50-C1 and HTB30-C11 are 5α-bufadienolides, HTB50-C2 and HTB30-C8 are pregnanes, HTB30-C7, HTB30-C12a, HTB30-C12b, HTB30-C13a, HTB30-C17 and HTB30-C19 are spirostanes, HTB30-C5 and HTB30-C14 are furostanes.Effects on catecholamine secretion in cultured bovine adrenal medullary cells were conducted on some isolated compounds. HTE-C2 and HTE-C5 enhanced catecholamine secretion, and these results indicated that the two compounds show excitability effects on sympathetic activity.The studies on chemical components and bioactivities of the Helleborus genus in the past years were also summarized.更多还原