多组分、代谢网络多靶点的PK-PD研究左金丸的药效物质基础及配伍机制

Zuojin wan is a classical formulae for the treatment of stomach heat syndrome (SH), but its compatibility mechanism and foundation of efficacy was yet not clear. After the effectiveness of zuojin wan on SH has been verified by using metabonomics, the aim of this study was to establish a new semi-quantitative method of obtaining the compositions of zuojin wan and its metabolites concentration-time curve(PK) and the endogenous metabolic biomarkers associated with SH concentration-time curve(PD) dynamically and synchronously in plasma of rats using UPLC-Q-TOF/MS by integration of serum pharmacochemistry, pharmacokinetics and metabonomics. The foundation of efficacy of zuojin wan was revealed by PK-PD correlation analysis and Prescription dismantlement study was further applied to explain the compatibility mechanism of cold and heat medicine joint use to treat SH from the following three aspects: PK of multi-components, metabolic profile and PD of multi-targets. Through comprehensive analysis of different information, the established method demonstrated the relationship among syndrome, constituents contained in blood, bioactive components and biomarkers, and had great significance for the interpretation of the compatibility mechanism and foundation of efficacy of Chinese compound formula, as well as the synergistic effects of multi-components. Meanwhile, the method provided an effective approach for the PK-PD correlation analysis of Chinese prescriptions.

左金丸是治疗肝火犯胃证的经典方剂，但配伍机制和效应物质基础却未完全清楚。在我们利用代谢组学确证了其对胃热证有效性的基础上，本项目拟融合血清药物化学、药代动力学及代谢组学的方法与技术，以UPLC-Q-TOF/MS为技术平台，以与胃热证相关内源性代谢标志物群为药效评价指标，建立一种能够动态、同步性获得大鼠血浆中左金丸所含成分及其代谢产物浓度-时间曲线（PK）、与胃热证相关内源性代谢标志物群浓度-时间曲线（PD）的半定量新方法，揭示药物体内量-时-效关系，通过PK-PD相关性分析揭示左金丸治疗胃热证的效应物质基础；拆方后在多组分PK、整体代谢轮廓与多靶点PD三个层面阐释其配伍机制。通过对不同信息的整合分析，本项目建立了证候-血中移行成分-药效组分-生物标志物的对应表观形式，对于阐释中药复方的配伍机制、药效物质基础及多组分协同增效作用具有十分重要的意义，同时为方剂PK-PD结合模型分析提供新思路。

左金丸是治疗肝火犯胃证的经典方剂，但其配伍机制和效应物质基础尚未完全清楚。项目以核磁共振（NMR）和超高效液相色谱串联三重四级杆飞行时间质谱（UPLC-Q-TOF-MS）代谢组学技术从大鼠尿液、粪便和血清分别鉴定了7个、5个和9个与胃热证相关潜在生物标志物，从整体性代谢网络层面阐释了胃热证的本质及左金丸的药效与机制。项目证实左金丸可通过抗炎、抗氧化应激和抗凋亡作用干预胃热证，为其临床合理用药提供了一定的理论基础。项目以UHPLC-Q-TOF-MS为技术平台，融合血清药物化学、药代动力学及代谢组学的思路与方法，建立了一种能够动态、同步性获得大鼠血浆中左金丸所含成分及其代谢产物浓度-时间曲线（PK）、与胃热证相关内源性代谢标志物群浓度-时间曲线（PD）的半定量新方法，通过PK-PD相关性分析发现黄连中的小檗碱、巴马亭、非洲防己碱、黄连碱、表小檗碱、羟基小檗碱和吴茱萸中的吴茱萸碱、吴茱萸次碱、去氢吴茱萸碱、Evocarpine可能为左金丸治疗胃热证的效应物质基础；相对于黄连和吴茱萸单独用药组，黄连和吴茱萸配伍应用（左金丸组）能显著提高效应物质基础的药时曲线下面积(AUC)、使胃热证大鼠血清整体代谢轮廓更接近正常组、能使大多数与胃热证相关潜在生物标志物产生显著性回调，这证明了黄连与吴茱萸寒热配伍并用治疗胃热证的合理性。项目实施建立的多组分、代谢网络多靶点的PK-PD结合分析方法通过对不同信息的整合分析，建立了证候-血中移行成分-药效组分-生物标志物的对应表观形式，为阐释中药复方的配伍机制、药效物质基础及多组分协同增效作用提供了可以借鉴的方法，同时，为成分复杂、疗效为多成分综合作用的中药复方PK-PD结合模型分析提供了一种有效的研究手段。