丁公藤的化学成分及生物活性研究
摘要
丁公藤(Erycibe obtusfolia Benth.)为旋花科(Convolvulaceae)丁公藤属植物。该属植物在我国共有11种,主要分布于广东、海南、广西、云南。丁公藤为我国传统中药,辛、温、有毒,具有祛风除湿,舒筋活络的功效。经现代药理实验证明具有缩瞳、抗炎镇痛等效用,临床用于治疗原发性青光眼、风湿痹痛、半身不遂、跌扑肿痛等病症。
本论文对丁公藤根的化学成分和生物活性进行了较系统的研究。丁公藤根经95%乙醇回流提取,萃取提取物,分为石油醚层、乙酸乙酯层、正丁醇层和水层。本文对乙酸乙酯层和水层进行了系统的化学成分研究,分别采用硅胶、大孔树脂、Sephdex LH-20和ODS柱色谱等方法进行分离,利用多种光谱方法(UV、IR、MS、~1H-NMR、~(13)C-NMR、2DNMR)对分离得到的化合物进行结构鉴定。分离并鉴定了16个化合物,包括7个香豆素类成分:7,7′-二羟基-6,6′甲氧基-3,3′-双香豆素(1),7,7′-二羟基-6,6′-二甲氧基-8,8′-双香豆素(2),7-O-[4′-O-(3″,4″-二羟基桂皮酰基)-β-D-吡喃葡萄糖基]-6-甲氧基香豆素(3),cleomiscosin A(8),cleomiscosin B(9),东莨菪素(10),东莨菪苷(11);5个绿原酸衍生物:3-O-4″-羟基-3″,5″-二甲氧基苯甲酰基绿原酸甲酯(4),4-O-4″-羟基-3″,5″-二甲氧基苯甲酰基绿原酸甲酯(5),灰毡毛忍冬素G(6),灰毡毛忍冬F(7),绿原酸(12);以及β-谷甾醇(13),胡萝卜苷(14),长链脂肪醇(15),胡萝卜苷连长链脂肪烷烃(16)。化合物1-5为新化合物,其中1、2为较为少见的碳碳键连接完全对称的双香豆素;6-9为首次从该属植物中分离得到。
对丁公藤的95%乙醇提取物和各萃取部位进行抗炎、抗HIV病毒和细胞毒活性筛选,结果表明:乙酸乙酯层和95%乙醇提取物对巴豆油致小鼠耳部炎症有明显的抑制作用,其抑制率分别为79.3%和40.0%;乙酸乙酯层对HIV病毒复制显示了较好的抑制作用,其抑制率为57.7%;石油醚层对卵巢癌细胞显示了很好的细胞毒活性,半数抑制浓度值(IC_(50))为12.65μg/ml。对化合物1,3-11进行抗炎的体外实验,观察化合物对小鼠腹腔巨噬细胞TNFα分泌的影响,其抑制率分别为6.2%、5.9%、6.3%、5.8%、15.8%、11.7%、9.7%、3.6%、9.3%和8.7%,均未表现出很好的活性。对化合物1,3-11进行抑制HIV病毒复制实验筛选,各化合物抑制率分别为26.6%、37.0%、0.0%、20.3%、20.7%、38.8%、0.0%、0.0%、18.1%和13.9%,其中,化合物3和7显示了较高的活性,抑制率分别为37.0%和38.8%。
Erycibe obtusifolia Benth.(Convolvulaceae) was distributed in the southern China.The genus Erycibe included 11 species in China.Its roots,stems and twigs were used in Chinese folk medicine to relieve the symptoms of rheumatoid arthritis. Pharmacological study indicated that Erycibe obtusifolia showed a strong anti -inflammatory activity and the effect of muscarinic agonist.
The constituents and bioactivities of Erycibe obtusifolia were studied.The dried roots of this plant were exhaustively extracted with 95%EtOH and concentrated under reduced pressure.Then the residue was suspended in H_2O and extracted successively with petroleum ether,EtOAc and n-BuOH.Sixteen compounds were isolated from the EtOAc and H_2O portions.Their structures were elucidated by spectroscopic methods including UV,IR,MS,HR-MS,1D and 2DNMR as 7,7'-dihydroxy-6,6'-dimethoxy-3,3'-biscoumarin(1),7,7'-dihydroxy-6,6'-dimethoxy -8,8'-biscoumarin(2),7-O-[4'-O-(3",4"-dihydroxycinnamyl)-β-D-glucopyranosyl]-6 -methoxycoumarin(3),methyl-3-O-(4"-hydroxy-3",5"-dimethoxybenzoyl)-chloro genate(4),methyl-4-O-(4"-hydroxy-3",5"-dimethoxybenzoyl)-chlorogenate(5), macranthion G(6),macranthoin F(7),cleomiscosin A(8),cleomiscosin B(9), scopoletin(10),scopolin(11),chlorogenic acid(12),β-sitosterol(13),daucosterol (14),alkyl alcohol(15),alkyl linked daucosterol(16).Among them,compounds 1-5 were new compounds,compounds 6-9 were isolated from genus Erycibe for the first time.Compounds 1 and 2 were the rare symmetrical coumarins coupled with carbon-carbon bonds.
All extractions were investigated and assayed anti-inflammatory activities, anti-HIV activities and cytotoxic activities.95%EtOH and EtOAc portions showed obvious anti-inflammatory activities in mice and their inhibition ratio were 79.3%and 40.0%respectively.EtOAc portion showed anti-HIV activities and the inhibition ratio was 57.7%.Petroleum ether portion showed cytotoxic activities with IC_((50)) 12.65μg/ml. In vitro the compounds 1,3-11 were assayed inhibitive activities of secretion of TNFα of macrophage and the inhibition ratio were 6.2%、5.9%、6.3%、5.8%、15.8%、11.7%, 9.7%、3.6%、9.3%and 8.7%respectively.The compounds 1,3-11 were assayed anti-HIV activities with inhibition ratio 26.6%、37.0%、0.0%、20.3%、20.7%、38.8%、0.0%、0.0%、18.1%and 13.9%respectively.The compound 3 and 7 showed higher inhibitory activities.
本论文对丁公藤根的化学成分和生物活性进行了较系统的研究。丁公藤根经95%乙醇回流提取,萃取提取物,分为石油醚层、乙酸乙酯层、正丁醇层和水层。本文对乙酸乙酯层和水层进行了系统的化学成分研究,分别采用硅胶、大孔树脂、Sephdex LH-20和ODS柱色谱等方法进行分离,利用多种光谱方法(UV、IR、MS、~1H-NMR、~(13)C-NMR、2DNMR)对分离得到的化合物进行结构鉴定。分离并鉴定了16个化合物,包括7个香豆素类成分:7,7′-二羟基-6,6′甲氧基-3,3′-双香豆素(1),7,7′-二羟基-6,6′-二甲氧基-8,8′-双香豆素(2),7-O-[4′-O-(3″,4″-二羟基桂皮酰基)-β-D-吡喃葡萄糖基]-6-甲氧基香豆素(3),cleomiscosin A(8),cleomiscosin B(9),东莨菪素(10),东莨菪苷(11);5个绿原酸衍生物:3-O-4″-羟基-3″,5″-二甲氧基苯甲酰基绿原酸甲酯(4),4-O-4″-羟基-3″,5″-二甲氧基苯甲酰基绿原酸甲酯(5),灰毡毛忍冬素G(6),灰毡毛忍冬F(7),绿原酸(12);以及β-谷甾醇(13),胡萝卜苷(14),长链脂肪醇(15),胡萝卜苷连长链脂肪烷烃(16)。化合物1-5为新化合物,其中1、2为较为少见的碳碳键连接完全对称的双香豆素;6-9为首次从该属植物中分离得到。
对丁公藤的95%乙醇提取物和各萃取部位进行抗炎、抗HIV病毒和细胞毒活性筛选,结果表明:乙酸乙酯层和95%乙醇提取物对巴豆油致小鼠耳部炎症有明显的抑制作用,其抑制率分别为79.3%和40.0%;乙酸乙酯层对HIV病毒复制显示了较好的抑制作用,其抑制率为57.7%;石油醚层对卵巢癌细胞显示了很好的细胞毒活性,半数抑制浓度值(IC_(50))为12.65μg/ml。对化合物1,3-11进行抗炎的体外实验,观察化合物对小鼠腹腔巨噬细胞TNFα分泌的影响,其抑制率分别为6.2%、5.9%、6.3%、5.8%、15.8%、11.7%、9.7%、3.6%、9.3%和8.7%,均未表现出很好的活性。对化合物1,3-11进行抑制HIV病毒复制实验筛选,各化合物抑制率分别为26.6%、37.0%、0.0%、20.3%、20.7%、38.8%、0.0%、0.0%、18.1%和13.9%,其中,化合物3和7显示了较高的活性,抑制率分别为37.0%和38.8%。
Erycibe obtusifolia Benth.(Convolvulaceae) was distributed in the southern China.The genus Erycibe included 11 species in China.Its roots,stems and twigs were used in Chinese folk medicine to relieve the symptoms of rheumatoid arthritis. Pharmacological study indicated that Erycibe obtusifolia showed a strong anti -inflammatory activity and the effect of muscarinic agonist.
The constituents and bioactivities of Erycibe obtusifolia were studied.The dried roots of this plant were exhaustively extracted with 95%EtOH and concentrated under reduced pressure.Then the residue was suspended in H_2O and extracted successively with petroleum ether,EtOAc and n-BuOH.Sixteen compounds were isolated from the EtOAc and H_2O portions.Their structures were elucidated by spectroscopic methods including UV,IR,MS,HR-MS,1D and 2DNMR as 7,7'-dihydroxy-6,6'-dimethoxy-3,3'-biscoumarin(1),7,7'-dihydroxy-6,6'-dimethoxy -8,8'-biscoumarin(2),7-O-[4'-O-(3",4"-dihydroxycinnamyl)-β-D-glucopyranosyl]-6 -methoxycoumarin(3),methyl-3-O-(4"-hydroxy-3",5"-dimethoxybenzoyl)-chloro genate(4),methyl-4-O-(4"-hydroxy-3",5"-dimethoxybenzoyl)-chlorogenate(5), macranthion G(6),macranthoin F(7),cleomiscosin A(8),cleomiscosin B(9), scopoletin(10),scopolin(11),chlorogenic acid(12),β-sitosterol(13),daucosterol (14),alkyl alcohol(15),alkyl linked daucosterol(16).Among them,compounds 1-5 were new compounds,compounds 6-9 were isolated from genus Erycibe for the first time.Compounds 1 and 2 were the rare symmetrical coumarins coupled with carbon-carbon bonds.
All extractions were investigated and assayed anti-inflammatory activities, anti-HIV activities and cytotoxic activities.95%EtOH and EtOAc portions showed obvious anti-inflammatory activities in mice and their inhibition ratio were 79.3%and 40.0%respectively.EtOAc portion showed anti-HIV activities and the inhibition ratio was 57.7%.Petroleum ether portion showed cytotoxic activities with IC_((50)) 12.65μg/ml. In vitro the compounds 1,3-11 were assayed inhibitive activities of secretion of TNFα of macrophage and the inhibition ratio were 6.2%、5.9%、6.3%、5.8%、15.8%、11.7%, 9.7%、3.6%、9.3%and 8.7%respectively.The compounds 1,3-11 were assayed anti-HIV activities with inhibition ratio 26.6%、37.0%、0.0%、20.3%、20.7%、38.8%、0.0%、0.0%、18.1%and 13.9%respectively.The compound 3 and 7 showed higher inhibitory activities.
引文
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