目的 研究中药复方热毒宁注射液解热抗炎活性部位的化学成分。方法 采用脂多糖（LPS）诱导小鼠内毒素休克模型对热毒宁注射液解热抗炎活性部位进行筛选，综合应用HP-20大孔树脂、硅胶、MCI、ODS柱色谱以及反相MPLC、HPLC等各种现代色谱技术对解热抗炎活性部位进行系统的分离纯化，根据化合物的光谱数据和理化性质进行结构鉴定。采用LPS诱导小鼠巨噬细胞RAW264.7模型，以抑制炎性细胞因子前列腺素E₂（PGE₂）的分泌为评价指标对分离得到的化合物进行体外抗炎活性评价。结果 热毒宁注射液经大孔吸附树脂95%乙醇洗脱部位为解热抗炎活性部位，从中分离得到了24个化合物，分别鉴定为 （4aS，7aS，7bS）-4，4a，7a，7b-tetrahydro-2H-1，7-dioxacyclopent[cd] indene-5-carboxylic acid methyl ester（1）、（4aS，7aS）-1，4a，5，7a-tetrahydro-7-（hydroxymethyl）-cyclopenta[c] pyran-4-carboxylic acid methyl ester（2）、3α，5α-tetrahydrodeoxycordifoline lactam（3）、R-（Z）-4-甲基-5-[（2'，6'，6'-三甲基-4'-氧代-2'-环己烯-1'-基） 亚甲基]-2（5H）-呋喃酮（4）、（1α，2α，3β，4β）-2，4-双 （4-羟基-3-甲氧基苯基）-1，3-环丁烷二甲酸（5）、4-[（6-O-苯甲酰基-β-D-吡喃葡萄糖基） 氧基]-3-甲氧基苯甲酸（6）、丁香脂素（7）、E-3-（3，4-二羟基苯亚甲基）-5-（3，4-二羟基苯基） 二氢呋喃-2-酮（8）、6，7-二甲氧基香豆素（9）、7-羟基-6-甲氧基香豆素（10）、水杨酸（11）、丁香醛（12）、苯乙酸（13）、香草醛（14）、咖啡酸（15）香草乙酮（16）、3，5-O-二咖啡酰奎宁酸（17）、4，5-O-二咖啡酰奎宁酸（18）、3，4-O-二咖啡酰奎宁酸甲酯（19）、3，5-O-二咖啡酰奎宁酸甲酯（20）、4，5-O-二咖啡酰奎宁酸甲酯（21）、5-O-咖啡酰奎宁酸乙酯（22）、3，5-O-二咖啡酰奎宁酸乙酯（23）、4，5-O-二咖啡酰奎宁酸乙酯（24）。其中化合物1、10、14~24 对LPS诱导的RAW264.7细胞中PGE₂的分泌具有显著的抑制作用。结论 化合物1~9、11~13、22~24 均为首次从热毒宁注射液中分离得到，有机酸类化合物可能为热毒宁注射液解热抗炎的主要药效物质基础之一。

Objective To study the antipyretic and anti-inflammatory constituents from the active fraction of Reduning (RDN) Injection. Methods In this study, the active fraction of RDN Injection was screened by the LPS-induced mouse endotoxin shock model. The chemical constituents were isolated by chromatography on HP-20 macroporous resin, silica gel, MCI, ODS, reverse MPLC, and HPLC repeatedly, and their structures were identified by spectral data and physicochemical property. Taking PGE₂ as the evaluating indicator, the model of LPS-induced RAW264.7 cells was used to evaluare the in vitro anti-inflammatory activity of these compounds. Results The 95% ethanol eluate of RDN Injection on the macroporous adsorption resin column was proved to be the antipyretic and anti-inflammatory active fraction of RDN Injection. A total of 24 compounds were isolated and identified as (4aS,7aS,7bS)-4,4a,7a,7b-tetrahydro-2H-1,7-dioxacyclopent[cd] indene-5-carboxylic acid methyl ester (1), (4aS,7aS)-1,4a,5,7a-tetrahydro-7-(hydroxymethyl)-cyclopenta[c] pyran-4-carboxy licacid methyl ester (2), 3α,5α-tetrahydrodeoxycordifoline lactam (3), R-(Z)-4-methyl-5-[(2',6',6'-trimethyl-4'-oxo-2'-cyclohexen-1'-yl) methylene]-2(5H)-furanone (4), (1α,2α,3β,4β)-2,4-bis(4-hydroxy-3-methoxyphenyl)-1,3-cyclobutanedicarboxylic acid (5), 4-[(6-O-benzoyl-β-D-glucopyranosyl) oxy]-3-methoxybenzoic acid (6), syringaresinol (7), E-3-(3,4-dihydroxybenzylidene)-5-(3,4-dihydroxyphenyl) dihydrofuran-2-one (8), 6,7-dimethoxy coumarin (9), 7-hydroxy-6-methoxy coumarin (10), salicylic acid (11), syringaldehyde (12), phenylacetic acid (13), vanillin (14), caffeic acid (15), aceto-vanillone (16), 3,5-di-O-caffeoylquinic acid (17), 4,5-di-O-caffeoylquinic acid (18), 3,4-di-O-caffeoylquinic acid methyl ester (19), 3,5-di-O-caffeoylquinic acid methyl ester (20), 4,5-di-O-caffeoylquinic acid methyl ester (21), 5-O-caffeoylquinic acid ethyl ester (22), 3,5-di-O-caffeoylquinic acid ethyl ester (23), and 4,5-di-O-caffeoylquinic acid ethyl ester (24). Among them, compounds 1, 10, and 14-24 significantly inhibited PGE₂ expression in RAW 246.7 cells stimulated by LPS. Conclusion Compounds 1-9, 11-13, and 22-24 are isolated from RDN Injection for the first time; And organic acids may be one of the main pharmacodynamic substances of RDN injection for antipyresis and anti-inflammation.

“重大新药创制”科技重大专项（2013ZX09402203）；热毒宁注射液标准化建设项目（ZYBZH-C-JS-31）