基于生物亲和色谱技术对芪苈强心胶囊干预慢性心力衰竭活性成分组与靶标群的关联研究

Compound traditional Chinese medicine is an integrated therapy characterized by ‘multi-compounds, multi-targets and multi-channels’. However, it’s pharmacodynamic material basis and mechanism are unclear, and the research methods are completely split with each other, namely ‘fragmentation’. Therefore, in this research Qiliqiangxin Capsules firstly recorded in the 2015 edition of Chinese Pharmacopoeia, which has definite effect on chronic heart failure verified by Evidence-based medicine, is used as a model to investigate the links between active compounds and target groups of Qiliqiangxin capsules intervening chronic heart failure. Firstly, the biological affinity chromatography is applied to systematically isolate, track and screen the active compounds and analyze their chemical structures. On that basis, the physiological disposition of active compounds in Qiliqiangxin Capsules and their corresponding dose-effect relationship are determined via PK-PD research. Lastly, the target groups are invented and predicted through network pharmacology, and the links between target groups and pharmacodynamic effects are analyzed. The integrated research model of ‘active compounds - physiological disposition - pharmacodynamic effects - network of target groups’ is eventually established by the complementary advantages of ‘biological affinity chromatography, PK-PD and network pharmacology’, so as to preliminarily clarify the material basis and mechanism of Qiliqiangxin Capsules intervening chronic heart failure, and to explore a feasible research idea to greatly improve the links between active compounds and target groups of Compound traditional Chinese medicine.

复方中药具有“多成分、多靶点、多途径”综合治疗疾病特点，但药效物质基础不明确、作用机制不清楚，且研究方法存在“碎片化”问题。为此，本课题采用经循证医学验证对慢性心力衰竭（CHF）疗效确切的复方中药芪苈强心胶囊为“范例”，研究芪苈强心胶囊干预CHF活性成分组与靶标群的关联性。首先通过生物亲和色谱对芪苈强心胶囊干预CHF的活性成分组进行系统分离、跟踪筛选、结构分析，在此基础上通过PK-PD研究芪苈强心胶囊中活性成分组的体内过程及其对应的量效关系，最后经网络药理学对活性成分组的靶标群进行虚拟、预测并解析与药物效应的相关性。通过“生物亲和色谱、PK-PD及网络药理学”研究方法的优势互补，建立复方中药“活性成分组-体内过程-药物效应-网络靶标群”整合研究模式，从而初步阐明芪苈强心胶囊干预CHF的药效物质基础和作用机制，探索出一条可行的且极大提高复方中药活性成分组与作用靶标群关联性的研究思路。

复方中药药效物质基础和作用机制研究是保证中药安全、有效和质量控制的前提，是国家中长期科学和技术发展规划纲要的一部分，是中医药走向现代化、国际化的核心。本研究采用生物亲和色谱、PK-PD及网络药理学研究方法，建立复方中药“活性成分组-体内过程-药物效应-网络靶标群”整合研究模式，以治疗慢性心力衰竭（CHF）疗效确切的复方中药芪苈强心胶囊为“范例”，研究芪苈强心胶囊干预CHF的药效物质基础和作用机制。研究按计划完成，⑴建立基于细胞膜色谱垂钓的复方中药专属性活性成分筛选方法，获得芪苈强心胶囊中能够与心肌细胞膜特异性结合的专属性活性成分4种（芒柄花素、丹酚酸B、丹酚酸A和槲皮素），与血管内皮细胞膜特异性结合的专属性活性成分5种（黄芪甲苷、丹酚酸B、丹酚酸A、羟基红花黄色素A和人参皂苷Rb1），与红细胞膜特异性结合的专属性活性成分1种（丹酚酸B），为复方中药药效物质基础研究提供思路；⑵建立PK比较研究方法探索疾病状态对活性成分吸收、代谢的影响，CHF病理状态下，芪苈强心胶囊中大部分活性成分呈现吸收延缓、吸收量和体内暴露量降低、清除延缓的特征，为临床合理用药提供提示；⑶建立基于药效活性成分的PK-PD相关性研究方法，提示中药复方药效产生是由多种活性成分联合作用的结果，同一药理作用在相同时间点出现的药效峰由相应的活性成分联合产生；而同一药理作用在不同时间点出现的药效峰是由产生各药效峰相同或不同的活性成分产生，这取决于相应的药效活性成分的PK行为，为复方中药“多成分、多靶点、多途径”的整体观提供新的诠释；⑷建立基于活性成分组的网络药理学分析方法，构建芪苈强心胶囊干预CHF的“活性成分组-疾病-靶标-生物过程/通路”网络，并通过人微血管内皮细胞（HCMEC）验证芪苈强心胶囊对同型半胱氨酸诱导的内皮细胞炎症损伤的保护作用，为阐释复方中药作用机制提供思路；⑸建立复方中药“活性成分组-体内过程-药物效应-网络靶标群”的整合研究模式，并应用该模式阐释芪苈强心胶囊干预CHF的药效物质基础以及作用机制。发表论文8篇，培养博士后和博、硕士研究生5名。