A Strategy For The Study Of ADME Process In Zebrafish Model Based On HRMS/MSn Dataset Mining Technique

Traditional Chinese medicine(TCM)is the cultural treasure of the Chinese nation.However,TCM usually contains hundreds of prototypes,which might be altered by the environmental or preparation factors and might undergo comprehensive biotransformation with diverse products.In addition to the complexity of biological matrices,the currently developed drugs research strategies in vivo absorption,distribution,metabolism,excretion(ADME)have great limitations and low analysis efficiency.Therefore,the research of potent material foundation of TCM has always been a hot and difficult issue in the modernization of TCM.In this paper,we have designed a highly efficient system for screening efficacy components of TCM with the integration of intelligent data acquisition,data mining,network pharmacology,and computer assisted target fishing,and has been successfully applied in Xiaokewan efficacy component screening in vivo rapidly.Firstly,with novel data acquisition(BE-DDA)and non-targeted data mining(PATBS)techniques,established an analytical process for rapid recognition and identification of unknown TCM components in complex biological matrices.53(34 substances screened in zebrafish and 19 in feeding solution)Xiaokewan related substances were successfully and quickly screened from zebrafish.And we found that the Xiaokewan main metabolite formononetin glucuronide conjugate was mainly distributed in the gastrointestinal tract and head of zebrafish by distribution study in vivo through mass spectrometry imaging.Then,the bioactive of 5 components(puerarin,daidzein,formononetin,deoxyschizandrin A and glybenclamide)with high exposure concentration in vivo was rapidly predicted by multiple computer assisted techniques such as systemic biology,network analysis and docking.Results shown that these five compounds could target signaling pathway related to diabetic syndrome such PI3K-Akt,type Ⅱ diabetes,TNF signaling pathway,insulin resistance and other signaling pathways to improve diabetes symptoms.And their pharmacological effects have been further verified by using diabetic zebrafish model.Meanwhile,the new established analysis process was also used to conduct a comprehensive analysis of Xiaokewan in rats,a total of 192 compounds(84 and 77 compounds exist only in plasma and urine,respectively)were detected.In comparison,found that the main exposed components of Xiaokewan in two animal models were very similar.This is the first time apply BEDDA and PATBS compounds identification technology and network pharmacological activity screening technology to screen rapidly the efficacy components of Xiaokewan in vivo.In summary,in this study,provides a new perspective to screen and investigate efficacy components of Xiaokewan,which provides a new perspective for the rapidly ADME research of TCM,and could serve as a referenced template for discovering the complex metabolism of TCM in vivo.