围绕“协同靶标组合”构建芪苈强心胶囊“一谱两效关联”的质量控制模式

"The uncorrelation between detection indexes and the effect” is a bottleneck problem for the quality control of traditional Chinese medicines (TCMs). In this project, Qiliqiangxin capsule, a clinically effective TCM for the treatment of heart failure, is the research object. Its potential pharmacological targets are beta adrenergic receptor and angiotensin converting enzyme, which constitute a synergetic target combination in the neuroendocrine system. Receptor biosensor and ultrafiltration-based enzyme extraction are two key technologies for the discovery of effective compounds. The research contents are as follows. (1) Establishment of the consistency evaluation method based on one spectrum (chemical fingerprint) and two effects (receptor biosensing curve and enzyme kinetics). (2) Screening of two effective molecular sets acting on beta adrenergic receptor and angiotensin converting enzyme respectively, and assay of the dose-effect relationship for each bioactive compound against its corresponding target. (3) Knockout/knockin of the effective molecular sets and establishment of their spectrum-effect relationships. (4) Qualitative and quantitative evaluation of the synergy between two effective molecular sets in a system model. Based on the chemistry and bioactivity of TCMs, this project aims to establish a new method for the consistency evaluation. A strong correlation between detection indexes and the effect can be constructed by dose-effect relationships, spectrum-effect relationships and synergistic effects. This project will be helpful to solve the bottleneck problem of TCM quality control, namely, the uncorrelation between detection indexes and the effect.

“检测指标与药效不关联”是目前中药质量控制的瓶颈问题。本项目以临床治心衰疗效确切的芪苈强心胶囊为模式药，围绕神经内分泌系统中的协同靶标组合（β肾上腺素能受体、血管紧张素转换酶），以受体传感器与超滤酶萃取为核心技术，开展如下研究：①建立基于一谱两效（化学指纹图谱、受体-配体互作曲线、酶动力学曲线）的中药质量一致性评价方法；②分别筛选作用于β肾上腺素能受体、血管紧张素转换酶的“药效成分子集”，测定成分与靶标间的“单体量效”关系；③敲出/敲入“药效成分子集”，结合生物检测建立“组分谱效”关系；④在系统模型中定性定量评估“药效成分子集”之间的“协同增效”作用。本项目旨在从中药的化学属性和生物属性建立新的质量一致性评价方法，以“单体量效-组分谱效-协同增效”链条建立检测指标与生物效应之间的强关联，突破传统质量控制模式“检测指标与药效不关联”的瓶颈。

“检测指标与生物效应不关联”是当前中药质量评价的瓶颈问题。本项目以芪苈强心胶囊为模式药，探索中药“谱效关联”的研究策略，主要研究内容包括以下四个方面。①建立了基于逐峰剪切和敲出色谱法的“单谱单效关联”策略。以逐峰剪切从芪苈强心胶囊中获得71个组分并发现其中23个组分具有显著的自由基清除活性。敲出色谱法显示敲出单一成分不会影响芪苈强心胶囊整体活性，揭示了芪苈强心胶囊极高的活性稳定性。丹酚酸类、咖啡酰基奎宁酸类、黄酮类被确定为其中的主要活性化合物，表明“不同类成分作用于同一靶点”是复方作用的模式之一。②建立了基于质谱和酶功能检测的“一谱两效关联”策略。通过高分辨、高灵敏质谱技术实现芪苈强心胶囊中化学成分的定性、定量分析，以酶功能检测开展双靶点α-葡萄糖苷酶/乙酰胆碱酯酶抑制剂的筛选。结果表明部分皂苷类成分选择性作用于α-葡萄糖苷酶，部分丹参酮类成分选择性作用于乙酰胆碱酯酶，表明“不同类成分选择性作用于不同靶点”是复方作用的模式之一。③初步探索基于质谱和网络药理学的“一谱多效关联”策略。结果显示芪苈强心胶囊中化学成分作用的靶点主要集中于神经活性配体-受体相互作用、钙信号通路、嘌呤代谢等通路，提示“化学成分-靶点交互网络”是复方作用的模式之一。④实现“单谱单效关联”与“一谱两效关联”的推广应用。“单谱单效关联”被推广应用于阔叶十大功劳和狭叶十大功劳，“一谱两效关联”被推广应用于陈皮、橘红、化橘红、枳实和枳壳，表明所建立的“谱效关联”系列策略具有普适性。本项目旨在从中药的化学属性和生物属性建立新的质量评价方法，以“单谱单效-一谱两效-一谱多效”链条建立检测指标与生物效应之间的强关联，突破传统质量控制模式“检测指标与生物效应不关联”的瓶颈。