基于血清谱效学和组合药代动力学的苓桂术甘汤药效物质基础及其配伍机理研究

"sovereign, minister, assistant and guide" is the core theory of the compatibility of traditional Chinese medicine (TCM), how to describe the compatibility rules based on pharmacodynamic material basis and mechanism of effect, and confirm the advantages of compatibility laws in toxicity attenuation and efficacy potentiation are scientific questions urgently needed to be solved. This project is on the basis of our early Natural Science Foundation research, and taking the classic TCM prescription named Ling-Gui-Zhu-Gan Decoction (LGZGD) as the research object: ① Using serum pharmacochemistry and metabolomic methods study the serum ingredients and their dynamic rules; ② Using "in vivo serum pharmacology" method which based on drug-containing serum reinjection technology to construct a "Adjusted Pharmacodynamics" (APD) mode; ③ Through the combination of "Serum Chromatographic Pharmacodynamics" (SCPD) and "combinatorial pharmacokinetics" (CPK) to clarify the pharmacodynamic material basis and their pharmacokinetic rules; ④ Based on the pharmacokinetic rules of the major pharmacodynamic material basis, qualitatively and quantitatively describe the synergy and antagonistic effect of the pharmacodynamic material basis of TCM prescriptions, clarify the compatibility rule of LGZGD, and expound the compatibility theories of TCMs further. This study intends to solve the key scientific issues are: Using pharmacokinetic rules interpreting the scientific connotation of the compatibility theories of TCMs, and frame a theory and technology platform for the research of pharmacodynamic material basis of TCMs.

君臣佐使是中药复方配伍理论的核心，如何从物质基础和作用机理层面研究君臣佐使配伍规律，证实复方配伍在减毒增效方面的优势，是丰富和发展中药复方配伍理论亟待解决的科学问题。本项目系在前期自然科学基金研究的基础上，以中药经典方剂"苓桂术甘汤"为研究对象：①应用血清药物化学与代谢组学研究血清效应成分谱及其动态规律；②应用基于含药血清回注技术的"在体血清药理学"方法，构建"校正药效动力学"（APD）模式；③通过"血清谱效学"（SCPD）与"组合药动学"（CPK）方法阐述中药复方的药效物质基础及其药动学规律；④根据主要药效物质基础的药动学规律，定性、定量描述中药复方药效物质基础的协同与拮抗作用，阐明经典方剂"苓桂术甘汤"中各药味的配伍规律，进而阐明中药复方君臣佐使的配伍理论。本研究拟解决的关键科学问题是：用药代动力学规律诠释中药复方君臣佐使配伍理论的科学内涵，构架中药复方药效物质基础研究的理论与技术平台

中药复方发挥疗效具有多靶点和多成分协同作用的整体特点，中药复方药效物质基础及其配伍规律的阐明是中药研究的基础。中药药效物质基础及其配伍规律的研究方法则是中药现代化和国际化进程的技术瓶颈，是国家“重大新药创制”科技重大专项计划资助的关键技术。只有建立中药药效物质基础及其配伍规律的系统研究方法，才能阐明中药及其复方的药效物质基础及其作用机理与配伍关系、建立有效的质量控制标准，进而加速中药现代化和国际化进程。 .本项目以张仲景的《伤寒论》名方苓桂术甘汤为研究对象，以急性心肌缺血模型大鼠为研究载体，以血清中肌酸激酶为药效学指标，建立药效动力学曲线；同时建立了“不加校正因子内参比法”的体内多组分高通量分析法，并构建了基于“表观效应浓度”的“组合药代动力学”研究模式，阐明了苓桂术甘汤治疗急性心肌缺血的药效物质基础及其配伍规律：①以代谢组学为技术手段评价并确立了“异丙肾上腺素”致大鼠急性心肌缺血的模型稳定性至少为三天；②利用高分辨质谱，对苓桂术甘汤的化学成分谱和给药后健康大鼠体内的血浆指纹谱进行全面分析，阐明方剂的入血成分谱；③通过比较计算模型大鼠给予苓桂术甘汤前后血清中肌酸激酶的变化规律，绘制了苓桂术甘汤治疗急性心肌缺血的药效-时间曲线；④建立了大鼠血浆中11种移行成分的高通量分析法；⑤通过“药效动力学-药代动力学拟合”，阐明苓桂术甘汤治疗急性心肌缺血的药效物质基础。⑥通过茯苓单体给药和茯苓-白术药对给药以及桂枝单体给药和桂枝-甘草药对给药的比较药动学的研究，阐明苓桂术甘汤君臣佐使的配伍理论和科学内涵。⑦依据浓度组合模型计算给药后各效应时间点血浆移行成分的整体“表观效应浓度”，并建立了“组合药代动力学-药效动力学”（CPK-PD）模型，评价了苓桂术甘汤治疗急性心肌缺血药效物质基础及其配伍规律的合理性，为中药药效物质基础的系统研究提供新的思路和方法。