International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development of Ministry of Education (MOE) of China/Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research/Institute of Traditional Chinese Medicine & Natural Products, College of Pharmacy, Jinan University, Guangzhou 510632, China.
Department of Pharmacology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
Phytomedicine. 2023 Jan;108:154511. doi: 10.1016/j.phymed.2022.154511. Epub 2022 Oct 22.
Quality marker (Q-marker) serves an important role in promoting the standardization of the quality of traditional Chinese medicine (TCM) prescriptions. However, discovering comprehensive and representative Q-markers from TCM prescriptions composed of multiple components remains difficult.
A three-step-based novel strategy integrating drug metabolism and pharmacokinetics (DMPK) with network pharmacology and bioactivity evaluation was proposed to discover the Q-markers and applied to a research example of Danlou tablet (DLT), a famous TCM prescription with remarkable and reliable clinical effects for coronary heart disease (CHD).
Firstly, the metabolic profile in vivo of DLT was systemically characterized, and the pharmacokinetic (PK) properties of PK markers were then investigated. Secondly, an integrated network of "PK markers - CHD targets - pathways - therapeutic effects" was established to screen out the crucial PK markers of DLT against CHD. Thirdly, the crucial PK markers that could exhibit strong myocardial protection activity in the H9c2 cardiomyocyte model were selected as the candidate Q-markers of DLT. According to the proportion of their C value in vivo, the candidate Q-markers were configured into a composition; the bioactivity was then evaluated to confirm their synergistic effect and justify their usage as Q-markers.
First of all, a total of 110 DLT-related xenobiotics (35 prototypes and 75 metabolites) were detected in bio-samples, and the pharmacokinetic properties of 13 PK markers of DLT were successfully characterized, revealing the quality transitivity and traceability from prescription to in vivo. Then, 6 crucial PK markers with three topological features (degree, betweenness, and closeness) greater than the average values in the pharmacology network were screened out as the key components of DLT against CHD. Furthermore, among these 6 crucial PK markers, 5 components (puerarin, alisol A, daidzein, paeoniflorin, and tanshinone IIA) with strong myocardial protection activity were chosen as the candidate Q-markers to constitute a new composition. The composition activated the expression of the PI3K/AKT pathway and exhibited strong myocardial protection activity, and the effective concentrations (nM level) of these components in the composition were significantly lower than their individually effective concentrations (μM level), indicating that there was a certain synergistic effect between them. Hence, the 5 components with multiple properties, including testability, quality transitivity and traceability from prescription to in vivo, effectiveness, and compatibility contribution, were defined as comprehensive and representative Q-markers of DLT.
This study not only presented a novel idea for the revelation of comprehensive and representative Q-markers in quality control research of TCM prescriptions, but also identified the reasonable Q-markers of DLT for the first time to improve the quality control level of DLT.
质量标志物（Q-marker）在促进中药（TCM）处方质量标准化方面发挥着重要作用。然而，从由多种成分组成的 TCM 处方中发现全面且有代表性的 Q-marker 仍然具有挑战性。
提出了一种三步策略，将药物代谢动力学（DMPK）与网络药理学和生物活性评估相结合，用于发现 Q-marker，并应用于丹络片（DLT）这一著名的 TCM 处方的研究实例，该处方对冠心病（CHD）具有显著而可靠的临床疗效。
首先，系统地描述了 DLT 的体内代谢谱，并研究了 PK 标志物的药代动力学（PK）特性。其次，建立了一个“PK 标志物-CHD 靶点-途径-治疗效果”的综合网络，筛选出 DLT 针对 CHD 的关键 PK 标志物。第三，选择在 H9c2 心肌细胞模型中具有强烈心肌保护活性的关键 PK 标志物作为 DLT 的候选 Q-marker。根据其体内 C 值的比例，将候选 Q-marker 配置成一个成分；然后评估其生物活性，以确认它们的协同作用，并证明它们作为 Q-marker 的使用合理性。
首先，在生物样本中总共检测到 110 种与 DLT 相关的外源性物质（35 种原型和 75 种代谢物），并成功地描述了 13 种 DLT 的 PK 标志物的 PK 特性，揭示了从处方到体内的质量传递和可追溯性。然后，筛选出 6 个具有三个拓扑特征（度、介数和紧密度）大于药理学网络中平均值的关键 PK 标志物，作为 DLT 治疗 CHD 的关键成分。此外，在这 6 个关键 PK 标志物中，选择了 5 个具有较强心肌保护活性的成分（葛根素、阿利松 A、大豆苷元、芍药苷和丹参酮 IIA）作为候选 Q-marker 来构成新的成分。该成分激活了 PI3K/AKT 通路的表达，并表现出强烈的心肌保护活性，且该成分在组成中的有效浓度（nM 级）明显低于其单独的有效浓度（μM 级），表明它们之间存在一定的协同作用。因此，这 5 个具有多种特性的成分，包括可测试性、从处方到体内的质量传递和可追溯性、有效性和配伍贡献，被定义为 DLT 的综合且有代表性的 Q-marker。
本研究不仅为揭示 TCM 处方质量控制研究中全面且有代表性的 Q-marker 提供了新的思路，还首次确定了 DLT 的合理 Q-marker，以提高 DLT 的质量控制水平。
