以具有肝毒性的中药反应性代谢产物为导向的中药配伍禁忌致/增毒机制研究

Drug bioactivation leading to the formation of reactive species capable of covalent binding to proteins represents an important cause of drug-induced toxicity. The object of this study was focus on the drug and anti-drug combinations in the eighteen incompatibility pairs of Veratrum nigrum and its drug incompatibility in TCM. The reactive metabolites were trapped with GSH-Br which was then analyzed by LC-MS/MS analysis followed by mass defect filtering and isotope pattern matching. The preparative liquid chromatography was used to separate chemical constituents from the multi-components system. After the validation of the toxic effect of the reactive metabolites, many multidisciplinary experiments were carried on to study the effect of anti-drug combination on the toxicity-effect characterization of the original drug. Furthermore, the reason of the increased toxicity-effect was illustrated by in vitro enzyme experiments. We systemically studied the relationship between eighteen incompatible medicaments and reactive metabolites, and this study was to provide experimental evidences to the mode of herb-herb interaction based on toxic metabolites and the mechanism of incompatible medicaments. All these results indicated that compatibility of medicines in a prescription also have interaction based on the metabolism. Consequently, this project will be not only helpful for the toxicity/ increase virulence mechanisms of the reactive metabolites, but also of value in enlarging the possible mechanisms ideas of the core content of the research of TCM incompatibility theory for providing a rationales for the safety of clinical practice.

药物诱导的毒性作用常来源于药物经代谢所产生的反应性代谢产物。本项目从药代动力学的角度出发，选择“十八反”中“诸参辛芍叛藜芦”作为模式配伍禁忌研究对象，采用溴代谷胱甘肽衍生化试剂亲核捕获配伍禁忌药材经代谢诱导后可能产生的谷胱甘肽加合物，依据同位素分子特殊质谱裂解规律逆推原型结构并分离提纯药材中的潜在肝毒性成分。在此基础上运用多学科技术手段有针对性的考察配伍反药对于体内外试验系统中所监测潜在肝毒性代谢产物的量－毒－时动态变化过程并进一步追踪这些影响的内在酶学机制，阐明配伍药材在治疗疾病过程中机体对其产生的影响及其与反药配伍之间的相互关系。本项目旨在揭示一类基于药物体内代谢过程的中药配伍禁忌研究，阐明由毒性反应性代谢产物所致中药配伍禁忌的分子机制。另外，本项目的研究方法可用来研究“十八反”以外的配伍禁忌药对，为中药配伍禁忌的研究提供新的策略和方法，对丰富和发展中药配伍禁忌理论系统有一定意义。