【作者】 吴斌；

【导师】 吴立军； 金哲史；

【作者基本信息】 沈阳药科大学 ， 药物化学， 2004， 博士

【摘要】 麻叶千里光(Senecio cannabifolius Less.)又名宽叶返魂草，为菊科(Compositae)千里光属(Senecio L.)多年生草本植物，主要分布于我国东北、河北等地及朝鲜、日本、前苏联远东地区，生长于山沟林缘及湿草甸上。本品味苦、性凉，具有散瘀、止血、止痛的功效，在民间被广泛用于病毒性感冒、各类炎症的治疗。以该植物为原料的中药制剂现有肺宁颗粒、返魂草注射剂等。该属植物的化学成分特征是含有吡咯里西啶类生物碱(pyrrolizidine alkaloids)，此类生物碱对人、家畜的肝脏具有很大的毒性，并且存在潜在的致癌危险。2004年美国、欧洲各药品监督管理部门对中国出口的同属植物千里光(S. scandens Buch.-Ham.ex D.Don)的各种制剂进行了封杀。为了解决本品显著疗效与较大毒性之间的矛盾，配合长春华康制药有限公司对该品种的二次开发，我们对麻叶千里光抗菌有效成分进行了较为详细的研究。 药效学实验结果表明，麻叶千里光水提取物具有较明显的抗菌、抗病毒作用，显著的解热、镇痛、抗炎、祛痰、止咳、平喘及增强免疫的作用，并有一定的抗过敏作用。其药效作用与临床作用一致。 本论文以革兰氏阳性菌金黄色葡萄球菌(Staphylococcus aureus)、枯草芽孢杆菌(Bacillus subtilis)和阴性菌大肠杆菌(Escherichia coli)为受试菌，采用体外抑菌圈法追踪抗菌活性部分，利用各种色谱技术，从麻叶千里光全草的水提醇沉物中，分离得到了52个化合物。根据理化性质和谱学数据鉴定了其中30个化合物的结构。其中新化合物有10个，分别为麻叶千里光碱A(Cannabiloid A, E-10)、麻叶千里光碱B(Cannabiloid B, B-6)、麻叶千里光碱C(Cannabiloid C, B-9)、麻叶千里光碱D(Cannabiloid D, B-10)、麻叶千里光苷A(Cannabiside A, E-15)、麻叶千里光苷B(Cannabiside B, B-2)、麻叶千里光苷C(Cannabiside C, B-5)、麻叶千里光苷D(Cannabiside D, B-13)、麻叶千里光苷E(Cannabiside E, B-20)、麻叶千里光内酯A(Cannabilactone A, C-4)；一个新天然产物：4-(吡咯烷-2-酮基)-苯基乙酸(4-(pyrrolidin-2-one)-phenyl acetic acid, C-1)。首次从千里光属植物(Senecio L.)中分离得到了14个化合物，它们分别是：对-羟基苯乙酸甲酯(methyl ρ-hydroxy ben-zeneacetate, E-3)、3，4-二羟基苯甲酸(3，4-dihydroxy benzoic acid, E-4)、2，5-沈阳药科大学博士学位论文中文摘要二经基苯乙酸(2，5一ihyd『OXyl benzeneaCetic aeid，E一5)、2，5一二轻基苯乙酸甲酷(mathyl一2，5劝hydroxy benzeneacetate，E一8)、3一轻基礴一甲氧基苯甲酸(3一hydroXy礴-mathoXy benzoie aeid，E一11)、5一轻基毗陡一2一甲酸甲醋(methyl一5一hydrOXy.2一Pyridine-ca由oXylate，E一13)、(6R，7as)6一HydroXy一7，7a一dihydro一2(6H)一benzofuranone(E一14)、2一(1，4一二轻基环己烷基)一乙酸(2一(1，4一dihydroxycyclohexanyl)一acetie aeid，E一16)、3一轻基环己酸(3一hydroxyeyelohexanecarboXylie aeid，E一17)、4一轻基苯甲醛(4一hydroxybenzaldehyde，B一14)、节基一O一奋L一口比喃阿拉伯糖基(1一6)井D一毗喃葡萄糖昔(benZyl6一O一全L一arabinoPyranosyl一声D一glucoPyranoside，B一16)、(6S，95)6一轻基一3一酮一a-紫罗兰醇一g一。声。一葡萄糖昔((65，95)9.0.声D习Iucopyranosy.oxy-6一hydroX梦3一OxO一企ionol，B一18)、(65，gR)6一轻基一3一酮一a-紫罗兰醇一9一O咨D一葡萄糖普((65，gR)9.0咨D一lu-eoPyranosyloXy一hydroxy-3一oxo一份ionol，B一19)、消旋丁香脂素((士)liriodendrin，C一2)。还有3个其他已知化合物:对一经基苯乙酸(P-hydroxy benzeneaCetic acid，E一1)、对-二苯酚(1，4一benzenediol，E一2)、邻轻基苯甲酸(o一hyd『OXy benzoic acid，E一9)、对经基苯甲酸(P-hydroXy benzoie aeid，E一12)、p一谷街醇(p一s如St6rol，C毛)。 体外抑菌实验表明，除水层外，各萃取部分均显抑菌活性，其中乙酸乙醋部分活性最强，能够显著抑制革兰氏阳性菌(金黄色葡萄球菌、枯草芽抱杆菌)，对革兰氏阴性大肠杆菌作用较弱。有效成分的分离鉴定结果显示，酚酸类物质是本品主要抗菌活性物质，利用比浊法测定化合物E一1~E一12的最小抑菌浓度(Mlc)，并对其构效关系进行了初步的讨论。 一般药理学研究显示，在高、中、低剂量下麻叶千里光提取物对试验动物中枢神经系统、心血管系统、呼吸系统无明显影响;急性毒性、长期毒性试验表明麻叶千里光提取物基本无毒。 本文采用均匀设计安排实验，通过高效液相色谱法和分光光度法测定麻叶千里光提取物中绿原酸、咖啡酸、寡糖的含量，以此作为指标，考察了粉碎程度、加水量、提取时间、提取次数、醇沉密度、醇沉浓度等6个因素对三者提取率的影响，实验结果用UROS软件系统和人工神经智能网络技术(art币eia!neura!ne七~O水，ANN)进行处理，优选出三者合理的提取工艺，为生产工艺参数的选择提供了实验依据。为了对该产品的生产进行质量监控，建立一个产品质量检验的方法，我们采用非水电位滴定法对本品的水提醇沉物?更多还原

【Abstract】 Doctor Candidate: Bin Wu Supervisor: Prof. Lijun WuProf. Chulsa KimSenecio cannabifolius Less., whose another name is "Kuan Ye Fan Hun Cao", as one species of Senecio genus of Compositae family, distributes mainly in the North east and Hebei Province of China, Korea, Japan and far east of pre-Soviet Union.The taste of this plant is bitter, and natured character is cool. While its function shows the effects on dispelling ecchymoma, anastalsis and analgesic. As one kind of folk medicine, it’s usually used to treat the diseases, such as virus influenza and many kinds of inflammatories.The character on its chemical constituents lies in the pyrrolizidine alkaloids, which exhibited the chronical hepatotoxicity (liver toxicity) and cancerigenic possibility to the human beings and domestic animals.In 2004, the importing of many kinds of preparations made from the species of Senecio genus from China had been prohibited by American and Europe drug administrative departments.In order to deal with the contradiction between the toxicity and efficiency on the treatment, to find the antibacterial compounds and made cooperation on the exploitation of this plant with Chang Chun Hua kang Pharm. Co. Ltd., the further research on the chemical constituents and their anti-virus activity were studied.Pharmacodynamics tests showed that the aqueous extract of Senecio cannabifolius Less, had the effect on antibiosis, antivirus, analgesic- antipyretic, anti-inflammatory antihechic, antianaphylaxis and immunoregulatory, etc., and also showed the same effect when it was used for clinical treatment.By many kinds of isolation method, such as silica gel chromatographies, Lobar column, semi-PHPLC and so on, 52 compounds were obtained and the structures of thirty of them were identified by their physico-chemical character and spectra, and comparison the data with the reference for them.Among the 30 compounds, ten of them were novel compounds. They were identified as Cannabiloid A (E-10), Cannabiloid B (B-6), Cannabiloid C (B-9), Cannabiloid D (B-10), Cannabiside A (E-15), Cannabiside B (B-2), Cannabiside C (B-5), Cannabiside D (B-13), Cannabiside E (B-20), Cannabilactone A(C-4). One new natural compound: 4-(pyrrolidin-2-one)-phenyl acetic acid (C-1). At same time, fourteen of them were isolated from the Senecio genus for the first time. They were elucidated as methyl p-hydroxy benzeneacetate (E-3), 3,4-dihydroxy benzoic acid (E-4), 2,5-dihydroxyl benzeneacetic acid (E-5), methyl 2,5-dihydroxy benzeneacetate (E-8), 3-hydroxy-4-methoxy benzoic acid (E-11), methyl-5-hydroxy-2-pyridinecarboxy-late (E-13), (6R,7aS)6-Hydroxy-7,7a-dihydro-2(6H)-benzofuranone (E-14), 2-(1,4-di-hydroxycyclohexanyl)-acetic acid (E-16), 3-hydroxycyclohexanecarboxylic acid (E-17), 4-hydroxy benzaldehyde (B-14), benzyl 6-O-a-L-arabinopyranosyl-B-D-glucopyrano-side (B-16), (6S,9S)9-O-B-D-glucopyranosyloxy-6-hydroxy-3-oxo-a-ionol (B-18), (6S, 9R)9-O-B-D-glucopyranosyloxy-6-hydroxy-3-oxo-a-ionol (B-19), (+)liriodendrin (C-2).By the anti-bacteria (screening) assay in vitro, the CHCI3, EtOAc and n-BuOH soluble fractions, except for the water layer also showed the potent anti-bacteria activities. Among them, the EtOAc layer exhibited the best result, which gave the potent activity against gram-positive bacilli, such as Staphylococcus aureus, Bacillus subtilis. By the chemical isolation, the main compounds of this part were phenolic constituents, and structure-activity relationship of compounds E-1~E-12 were discussed simply.The general pharmacological studies indicated that the high, medium and low dose of the extract had no influence on the central nervous system, cardiovascular and respiratory system. At same time, the tests of the extract on acute toxicity and secular toxicity showed no toxicity.Contents, such as chlorogenic acid, caffeic acid and oligosaccharides from the extrat of Senecio cannabofolius Less, were examined by HPLC and spectrophoto-metry with uniform design, and six factors, such as the extent of crush, amount of water,更多还原