基于特征图谱、化学计量学和分子对接的复方阿胶浆质量标志物研究  

Study of quality markers of Fufang E’jiao Jiang based on characteristic spectrum,chemometrics and molecular docking

作　　者：许啸 张淹[2] 任雪阳 王宇 董英 宋若兰 于啊香 魏静 马嘉慕 折改梅[1] XU Xiao;ZHANG Yan;REN Xue-yang;WANG Yu;DONG Ying;SONG Ruo-lan;YU A-xiang;WEI Jing;MA Jia-mu;SHE Gai-mei(School of Chinese Pharmacy,Beijing University of Chinese Medicine,Beijing 100102,China;National Engineering Research Center for Gelatin-based Traditional Chinese Medicine,Dong-E-E-Jiao Co.,Ltd.,Liaocheng 252201,China)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]山东东阿阿胶股份有限公司国家胶类中药工程技术研究中心,山东聊城252201

出　　处：《中草药》2021年第23期7148-7161,共14页Chinese Traditional and Herbal Drugs

基　　金：国家重点研发计划(2018YFC1707300);国家重点研发计划项目(2018YFC1706300)。

摘　　要：目的以质量标志物(quality markers,Q-Marker)"五原则"为指导,结合特征图谱、化学计量学、分子对接和含量测定等技术,开展复方阿胶浆(Fufang E’jiao Jiang,FEJ)治疗贫血症的Q-Marker研究。方法色谱柱Agilent Zorbax SB AQ-C18(250 mm×4.6 mm,5μm),流动相为0.1%甲酸乙腈溶液-0.1%甲酸水溶液,体积流量1.0 mL/min,柱温30℃,检测波长330 nm,焦地黄苯乙醇苷A1为参照峰,建立FEJ特征图谱。采用层次聚类分析(hierarchical clustering analysis,HCA)、主成分分析(principal component analysis,PCA)、偏最小二乘法-判别分析(partial least square method-discriminant analysis,PLS-DA)对不同批次FEJ进行分析。查阅《中国药典》和文献,结合组方配伍环境(君、臣、佐使)、成分传递与溯源等多因素,收集筛选质量控制6个关键成分。在GEO数据库中检索筛选贫血症(anemia)的差异基因,采用Discovery Studio 2016v16.1软件,将成分与贫血症相关靶点进行对接验证。以330、270 nm双波长同时测定FEJ中异毛蕊花糖苷和党参炔苷的含量。结果FEJ特征图谱中指认了咖啡酸、绿原酸、洋地黄叶苷C、阿魏酸、焦地黄苯乙醇苷A1、毛蕊花糖苷、焦地黄苯乙醇苷B1、异毛蕊花糖苷共8个特征成分。HCA、PCA和PLS-DA将16批FEJ分为3类,其中异毛蕊花糖苷、洋地黄叶苷C和焦地黄苯乙醇苷A1对FEJ质量影响最为显著。8个特征成分和6个关键成分的分子对接结果表明,异毛蕊花糖苷、党参炔苷、绿原酸、毛蕊花糖苷4个成分能够与激肽原-1(kininogen-1,KNG1)、泛素样蛋白ISG15(ubiquitin-like protein ISG15,ISG15)、赖氨酸特异性脱甲基酶6A(lysine-specific demethylase 6A,KDM6A)、乙酰化酶动力蛋白-3(dynamin-3,DNM3)、蛋白酶激活受体2(proteinase-activated receptor 2,F2RL1)、半胱氨酰白三烯受体1(cysteinyl leukotriene receptor 1,CYSLTR1)6个靶蛋白通过氢键、疏水键、π-π键等作用力结合,具有较好的活性,可作为FEJ的Q-Marker。16批Objective Guided by the“Five Principles”of quality markers(Q-Marker),combined with characteristic spectrum,chemometrics and molecular docking and content determination,developed Q-Marker for the treatment of anemia with Fufang E'jiao Jiang(复方阿胶浆,FEJ).Methods The characteristic spectrum of FEJ was determined by HPLC using Agilent Zorbax SB AQ-C18 column.The mobile phase was 0.1%formic acid acetonitrile-0.1%formic acid water,and the flow rate was 1.0 mL/min,detection wavelength was 330 nm.Sixteen batches of FEJ characteristic spectrum were established with jionoside A1 as the reference peak.Based on the results of common peak area determination of characteristic spectrums,the quality of different batches of FEJ was evaluated by hierarchica cluster analysis(HCA),principal components analysis(PCA)and partial least squares-discrimination analysis(PLS-DA).Six key components were collected from each herb in Pharmacopoeia of the People’s Republic of China and literatures.The differential genes of anemia was searched in the GEO database,and Discovery Studio software was used to verify the components in FEJ.The content of two components in 16 batches was determined at the same time at 330 nm and 270 nm.Results There were eight components were identified,namely caffeic acid,chlorogenic acid,purpureaside C,ferulic acid,jionoside A1,acteoside,jionoside B1,isoacteoside.The results of HCA,PCA and PLS-DA showed that 15 batches of FEJ can be divided into three categories.Isoacteoside,purpureaside C,and jionoside A1 were contributing to the quality difference between different batches of FEJ.The results of molecular docking showed that isoacteoside,lobetyolin,chlorogenic acid,and acteoside can bind to six key targets such as KNG1,ISG15,KDM6A,DNM3,F2RL1,and CYSLTR1 through hydrogen bonds,hydrophobic bonds,π-πbonds,etc.They can be used as Q-Markers of FEJ.The contents of isoacteoside and lobetyolin in 16 batches of FEJ were 6.05—12.30μg/mL and 9.14—16.30μg/mL,respectively.Conclusion Q-Markers in FEJ was explo

关 键 词：复方阿胶浆 质量标志物 特征图谱 化学计量学 分子对接 贫血症 层次聚类分析 主成分分析 偏最小二乘法-判别分析 质量控制 咖啡酸 绿原酸 洋地黄叶苷C 阿魏酸 焦地黄苯乙醇苷A_(1) 毛蕊花糖苷 焦地黄苯乙醇苷B_(1) 异毛蕊花糖苷 

分 类 号：R283.6[医药卫生—中药学]