整合PK/PD与代谢组学的黄连解毒汤体内药效物质基础研究

This project will clarify the in vivo pharmacodynamic material basis of the Traditional Chinese medicine (TCM) prescription, Huang-Lian-jie-Du-Decoction (HLJDD), for its function of “Clearing away Heat and Toxins” and treatment of stroke by an integration of PK/PD and metabolomics (PPMs), utilizing state-of-the-art techniques such as UPLC-Q-TOF-MS and NMR for qualitative and quantitative analysis of complex system, combined with biological means. Centering its efficacy in severe infection and stroke, taking the components of the formula as a whole, results of the pharmacokinetics (PK) and metabolomics will be interconnected via statistical models, profiling both the exogenous chemical components in the body and endogenous metabolites corresponding to the best efficacy. Those endogenous metabolites with differential ability could be retrieved to targets, based on which, effect networks were constructed. Combinations of component herbs, chemical compositions, and single compounds according to rational design of experiments (DOE) will be studied for their potency based on the constructed metabolomics profile to find better combinations. Such an idea of from effective ingredients to endogenous metabolites and to target networks is a feasible strategy of original innovation to the study of the material base of TCM formula that fully address its multi-component and multi-target characteristics, by rationally simplified the complexity of the two complicated systems of the chemical space of TCM formula and the body.

本项目整合药物代谢动力学/药效动力学/代谢组学（PPMs）技术，应用UPLC-Q-TOF-MS、NMR这些复杂体系的定性定量分析技术和生物学手段，对中药经典方剂黄连解毒汤传统清热解毒功效和近年发现的治疗脑卒中作用的体内药效物质基础进行研究。将方剂的化学成分作为一个整体，分别以抗重症感染和治疗脑卒中的药效学为核心，在大鼠体内将药物代谢动力学与代谢组学相关联，建立统计学模型，确定具有最佳药效时体内化学成分的组成和对应的内源性小分子代谢物谱，再根据各种内源性小分子代谢物反向推导相应的靶点群，并据此分别绘制出作用靶点网络图。组合配伍不同单味药、化学成分群、单体化合物，借助相应的内源性代谢物谱寻找具有更佳药效的物质组成。这种药效成分→内源性小分子代谢物→靶点群的思路与方法使对中药方剂多成分、多靶点的研究成为可能，合理简化了对中药方剂化学成分与机体这两个复杂体系相互作用研究的复杂程度，具有原始创新性。

黄连解毒汤是经典的清热解毒方剂，现代临床数据表明其对脓毒症及脓毒症引起的多器官功能障碍具有显著疗效。本项目建立了脓毒症和脑卒中两个动物模型，分别阐明黄连解毒汤治疗这两种疾病的物质基础。以经典的药效学（PD）指标作为显示疗效的标准，当达到最佳治疗效果时，研究其体内相应的药效物质基础，分析血清中所含化合物的种类及含量（PD）。另一方面确定动物表现出最佳药效的内源性小分子代谢产物的组成图谱（Metabolomics），从而将药代动力学/药效动力学/代谢组学（PPMs）进行整合，确定了具有最佳疗效的物质基础。我们还考察了黄连解毒汤原方及其拆方和主要单体小檗碱、黄芩苷、栀子苷对脓毒症和脑卒中大鼠的代谢组学研究，确定了各组之间内源性小分子代谢谱，阐明了其脓毒症和脑卒中的作用机制。另外，阐明了4-辛基衣康酸通过抑制糖酵解发挥抗炎作用的机制，该研究对4-辛基衣康酸调控代谢重编程提出了新颖的见解，并强调了靶向有氧糖酵解在治疗炎性疾病中的重要性，这为今后抗炎新药的研发提供了新的思路。本项目共发表标注SCI论文19篇，其中在国际知名学术期刊Nature Communications（IF：11.878）发表研究论文一篇，在Pharmacological Research (IF：5.57)发表一篇，在Aging and disease, Oxidative Medicine and Cellular Longevity， Frontiers in Pharmacology等国际著名期刊 5 篇。培养博士研究生3名，硕士研究生4名。通过本项目的研究，我们建立了与中药方剂多组分特点相适应的体内活性成分、药效学与内源性小分子之间的PPMs模型，能更好地阐明中药多组分作用的整体特征。这种PK和PD研究结果的模型拟合对中药方剂复杂体系的研究具有重要的指导意义。