热毒宁注射液化学成分研究(Ⅲ)  

Research on chemical constituents from Re-Du-Ning Injection(Ⅲ)

作　　者：李海波[1,2] 于洋[3] 王振中[1,2] 孟兆青[1,2] 房卉[1,2] 丁岗[1,2] 黄文哲[1,2] 萧伟[1,2] 姚新生[3] 

机构地区：[1]江苏康缘药业股份有限公司,江苏连云港222001 [2]中药制药过程新技术国家重点实验室,江苏连云港222001 [3]暨南大学中药及天然药物研究所,广东广州510632

出　　处：《中草药》2016年第10期1643-1649,共7页Chinese Traditional and Herbal Drugs

基　　金：江苏省自然科学青年基金:基于活性成分追踪分离及快速检识分析新策略的中药复方药效物质基础研究(BK20140441);江苏省博士后科研资助计划(1401171C)

摘　　要：目的研究热毒宁注射液抗病毒活性部位的化学成分。方法采用流感病毒致应激负荷小鼠模型进行抗病毒活性部位的筛选,综合应用硅胶、ODS、Sephadex LH-20和Toyopearl HW-40柱色谱以及反相MPLC、HPLC等现代色谱技术对抗病毒活性部位进行分离和纯化,根据化合物的光谱数据和理化性质鉴定化合物结构。通过采用检测海肾荧光素酶(Rluc)报告基因的表达水平来反映病毒感染水平的方法,对分离得到的化合物进行了体外抗登革热病毒的活性测试。结果热毒宁注射液经HP-20大孔吸附树脂,95%乙醇洗脱部位为抗病毒活性部位,从中分离得到14个化合物,分别鉴定为(2E,6S)-8-[α-Larabinopyranosyl-(1″-6′)-β-D-glucopyranosyloxy]-2,6-dimethylct-2-eno-1,2″-lactone(1)、落叶脂素(2)、5′-甲氧基异落叶松树脂醇(3)、异落叶松脂素(4)、(7R,8R)-4,7,9,9′-tetrahydroxy-3,3′-dimethoxy-8-O-4′-neolignan(5)、(7S,8R)-4,7,9,9′-tetrahydroxy-3,3′-dimethoxy-8-O-4′-neolignan(6)、肥牛木素(7)、5′-甲氧基肥牛木素(8)、(-)-二氢脱氢二松柏醇(9)、(7S,8R)-3,3′,5-trimethoxy-4′,7-epoxy-8,5′-neolignan-4,9,9′-triol(10)、2-顺式-4-反式-脱落酸(11)、2-反式-4-反式-脱落酸(12)、(1S,3R,4R,5S,7R,9R)-decane-6-carboxylic acid(13)、(1S,3R,4S,5S,7R,9R)-decane-6-carboxylic acid(14);其中化合物1对登革热病毒的EC50值为74.76μmol/L,选择指数(SI)值大于1.07,具有体外抗登革热病毒的活性。结论化合物8为新化合物,其他化合物均为首次从中药复方热毒宁注射液中分离得到,化合物1具有体外抗登革热病毒的活性。Objective To study the antiviral constituents from the active fraction of Re-Du-Ning（RDN） Injection. Methods In this study, the active fraction of RDN Injection was screened by the mice model loaded with restraint stress infected with influenza virus. The chemical constituents were isolated by chromatography on silica gel, ODS, Sephadex LH-20 Toyopearl HW-40 columns, and reverse phase MPLC HPLC repeatedly. Their structures were identified by spectral data and physicochemical property. Results The 95% ethanol eluate of RDN Injection on the macroporous adsorption resin column was proved to be the antivirus active fraction of RDN Injection. Fourteen compounds were isolated and identified as（2E,6S）-8-[α-L-arabinopyranosyl-（1″-6′）-β-D-glucopyranosyloxy]-2,6-dimethylct-2-eno-1,2″-lactone（1）, lyoniresinol（2）, 5′-methoxyisolariciresinol（3）, ent-isolariciresinol（4）,（7R,8R）-4,7,9,9′-tetrahydroxy-3,3′-dimethoxy-8-O-4′-neolignan（5）,（7S,8R）-4,7,9,9′-tetrahydroxy-3,3′-dimethoxy-8-O-4′-neolignan（6）, ceplignan（7）, 5′-methoxyceplignan（8）,（-）-dihydrodehydrodiconiferyl alcohol（9）,（7S,8R）-3,3′,5-trimethoxy-4′,7-epoxy-8,5′-neolignan-4,9,9′-triol（10）,（2-cis, 4-trans）-abscisic acid（11）,（2-trans, 4-trans）-abscisic acid（12）,（1S,3R,4R,5S,7R,9R）-decane-6-carboxylic acid（13）, and（1S,3R,4S,5S,7R,9R）-decane-6-carboxylic acid（14）; Among them, compound 1 exhibited the antivirus activity against Dengue virus. Conclusion Compound 8 is a new compound and the other isolated compounds are reported from RDN Injection for the first time, and compound 1 shows the anti-virus activity against Dengue virus.

关 键 词：热毒宁注射液 5′-甲氧基肥牛木素 木脂素 落叶脂素 2-顺式-4-反式-脱落酸 抗登革热病毒活性 

分 类 号：R969[医药卫生—药理学]