基于潜在药效物质组的中药注射剂PK-BN-PD模型的构建与应用

Based on a scientific issue that the quality control components of traditional Chinese medicine（TCM）is difficult to characterize the overall efficacy of TCM, the concept of the potential efficacy material markers (EMM) was introduced, and EMM mainly included prototype components and their metabolites groups which participating efficacy of TCM. Compared with traditional methods, this method has the advantage in quickly and accurately determining on quality control components from complex system after administration of TCM..In this study, Kudiezi injection was chosen as research vector. On the basis of preliminary chemical constituents and pharmacological studies, the first application of Bayesian networks (BN) technology to build TCM metabolism/effect (PK/PD) model. The research was developed for screening the EMM of Kudiezi injection intervention focal cerebral ischemia reperfusion and for screening quality control components basing of EMM and multi-component interactions in vivo. UPLC-MS/MS was applied to determination of multi-component concentrations in MCAO and reperfusion rat brain tissue or in blood. Changes in concentration by inflammatory cytokines and neurotransmitters as efficacy indicators, the innovation strategy of PK-BN-PD screening quality control indicators mode was developed based on EMM. Complete the task will provide a reference model to quickly identify indicators of quality control of TCM based on the efficacy of TCM and provide new ideas for the establishment of the quality evaluation system for TCM.

本课题针对中药质量控制指标难以表征中药整体药效的科学问题，创新性地提出潜在药效物质组（EMM）的概念，用以表征参与中药整体药效的中药原型成分及其代谢物质组。与传统方法比较，这种方法具有在整体/复方给药基础上从复杂体系中快速准确确定中药质量控制指标的优势。.本课题首次应用贝叶斯网络（BN）技术构建中药代谢/效应（PK/PD）模型，开展苦碟子注射液干预大鼠脑缺血再灌注的EMM的筛选以及基于EMM和体内多成分相互作用的质量控制指标的筛选研究。应用UPLC-MS/MS测定MCAO再灌注模型大鼠血液或脑组织中苦碟子多成分的浓度，以炎症细胞因子和神经递质的浓度变化作为药效指标，构建基于PK-BN-PD筛选苦碟子注射液的EMM和质量控制指标的新策略。课题的完成，将为基于中药整体药效的中药质量控制指标的快速确定提供参考模式，为中药质量评价体系的建立提供新的思路。

本课题针对中药质量控制指标难以表征中药整体药效的科学问题，创新性地提出潜在药效物质组（EMM）的概念，用以表征参与中药整体药效的中药原型成分及其代谢物质组。与传统方法比较，这种方法具有在整体/复方给药基础上从复杂体系中快速准确确定中药质量控制指标的优势。.本课题围绕关键问题，课题组首先对苦碟子注射液及其药材研究的文献进行了较为全面的梳理和综述。采用HPLC-DAD-ESI-MS/MS以及UHPLC-LTQ-Orbitrap-MS两种检测方法完成对苦碟子药材及其注射液中134个化学成分的表征。构建了系列UPLC-ESI-MS/MS和HPLC-ELSD方法，对19个苦碟子注射液中的主要成分作为质量控制的指标性成分进行了含量测定研究。应用UHPLC-LTQ-Orbitrap-MS对苦碟子注射液体内、体外的原型及代谢产物即药源性物质组进行了鉴定：大鼠血浆中分析检测38个、在健康大鼠脑组织中鉴定5个、模型大鼠脑组织中鉴定16个、心肌组织中鉴定26个；鉴定了苦碟子注射液中倍半萜内酯类成分经肝微粒体代谢的17个原型及代谢产物。建立“Cocktail”探针法结合UPLC-MS/MS定量分析方法考察苦碟子注射液及倍半萜内酯类化合物（IZ、DIZ）对大鼠肝细胞色素P450酶活性的抑制作用。建立并验证了一系列同时测定体内苦碟子注射液十多个成分血药浓度的灵敏快速的UPLC-MS/MS方法，并应用于苦碟子注射液在大鼠血浆、脑组织、心肌组织、犬血浆的药代动力学研究；同时应用数据挖掘技术构建苦碟子注射液代谢/效应（PK/PD）模型，开展了基于多成分药代动力学与药物整体效应的苦碟子注射液潜在药效物质组与质控指标的筛选研究。该项目的顺利实施和基于PK-PD筛选EMM和质量控制指标的新策略的构建，为基于中药整体药效的中药质量控制指标的快速确定提供参考模式，为中药质量评价体系的建立提供新的思路。.