板蓝根中芥子苷类成分的体内过程与PK-PD研究

Chirality is a basic feature of biological systems. Given oral administration of Traditional Chinese Medicine, chiral chemical components contained may show stereoselective differences in pharmacokinetics and pharmacodynamics. Thus the in vivo process of chiral chemical components of TCM and its correlation with the efficacy are worth of further research..This study was based on the theory of Comparative Traditional Chinese Medicine, targeting glucosinolates (progoitrin and epiprogoitrin，S-goitrin and R-goitrin), characteristic active components of Isatidis Radix, and studying on the chiral glucosinolates in vivo biotransformation and pharmacokinetics. By using the method and technology of pharmacokinetics, chemistry, analytical chemistry and pharmacology of TCM, comparative study on the in vivo process of glucosinolates was carried on, and anti-endotoxin effect of prototype and its metabolites was discussed. Comparison of glucosinolates in vivo pharmacokinetic characteristics between normal and febrile rats and the application of pharmacokinetic-pharmacodynamic model were in place to illuminate the relationship between blood drug concentration-time-effect obtained from prototype and its metabolites, and to state glucosinolates in vivo biotransformation as well as their efficacy material foundation, which laid groundwork for further study on the mechanism of Radix Isatidis. This study was a significant reference both in theory and in practice to make an approach for other TCM which contain similar chiral chemical components.

手性是生物系统的基本特征，中药口服后其中所含的手性活性成分可能会出现药动学和药效学的立体选择性差异，因此中药手性成分的体内过程及其与药效的相关性值得深入研究。.本项目以比较中药学为总体思路，以板蓝根中的特征性活性成分芥子苷类（原告依春和表原告依春、S-告依春和R-告依春）为研究对象，以手性芥子苷类成分的体内生物转化和药代动力学特征为核心内容，运用中药药代动力学、中药化学、分析化学和中药药理学的技术和方法，对芥子苷类的体内过程以及原型和主要代谢产物的抗内毒素作用进行比较研究；对芥子苷类在正常和发热大鼠体内的药代动力学特征进行比较，并应用药动学-药效学结合模型，阐明原型和主要代谢产物的血药浓度-时间-效应之间的关系，阐明芥子苷类成分的体内生物转化过程和药效物质基础，为进一步研究板蓝根的作用机制奠定基础。本研究具有理论价值和现实意义，对于其它类似含手性活性成分的中药研究亦有借鉴意义。

本项目对板蓝根进行化学成分及其抑制一氧化氮释放活性、LPS诱导内毒素败血症小鼠的保护作用及机制、芥子苷类的体内过程及其在正常和发热大鼠体内的药代动力学特征进行研究。.板蓝根经80%乙醇提取3次，浓缩，用石油醚、二氯甲烷和正丁醇依次萃取，经多种色谱方法系统分离纯化，结合理化性质和波谱学数据等方法鉴定结构，LPS诱导产生的RAW264.7细胞测定化合物的NO抑制活性。共分离并鉴定了146个单体化合物，其中34个新化合物，8个新天然产物，41个化合物为首次分离，25个化合物具有潜在的抗炎活性。.采用LPS诱导的细菌内毒素休克模型小鼠和RAW264.7细胞为实验模型，探究板蓝根体内外抗内毒素的药效作用及机理。板蓝根对LPS诱导内毒素小鼠具有保护作用，其调控IRF3下游炎症指标IFN-β的转录水平及释放，并阻断IFN-β/STAT干扰素信号传导通路，抑制其下游促炎细胞因子CXCL9、CXCL10、CXCL11的转录表达，从而发挥保护小鼠脏器组织，减缓内毒素引起的机体损伤。.采用超高效液相色谱-四级杆/飞行时间串联质谱（UHPLC-Q/TOF-MS）研究芥子苷类的体内过程。建立血浆样本中R,S-告依春、原告依春和表原告依春的LC-MS/MS分析方法，并计算其在大鼠体内的药动学参数。进一步检测R,S-告依春在正常和酵母致热大鼠体内的血药浓度，并计算药代动力学参数，与正常大鼠进行比较。R,S-告依春在大鼠体内的代谢途径主要涉及甲基化、乙酰化、羟基化、氧化、还原反应、与谷胱甘肽结合和与丙氨酸结合，环状结构稳定不易被水解。R,S-告依春口服摄入体内后Cmax与AUC值皆呈剂量依赖性，但酵母致热组的Cmax与AUC值明显高于正常组，且酵母致热组中R,S-告依春半衰期延迟，血浆清除速度减慢，平均滞留时间增加，提示发热病理状态下会影响R,S-告依春的药代动力学行为。原告依春和表原告依春口服摄入体内后经胃肠道迅速吸收，呈剂量依赖性，两者药代动力学特征相似，表原告依春的最高血浆浓度（Cmax）和AUC值略高于原告依春。而静脉内给药时两者的药代动力学行为明显不同。表原告依春的Cmax、AUC值以及半衰期比原告依春约高3倍。原告依春的口服生物利用度为20.1％–34.1％，是表原告依春的3倍。.本研究具有理论价值和现实意义，为进一步研究板蓝根奠定基础，对其它类似含手性活性成分的中药研究亦有借鉴意义。