Investigation On NMR Cell Metabonomics Of Apatinib And Coumarin Derivatives

Metabonomics is used to study the metabolites characteristics of cells,tissues,organs,or the whole organism.Among them,the NMR-based metabonomics is one of the most important analysis methodsIn this paper,the NMR metabonomics was used to analyze the metabolic pathway of apatinib in inhibiting the growth and proliferation of HepG2 cells.The anti-tumor activities of 12 novel coumarin derivatives were preliminarily studied.In the first part,the mechanism and metabolic pathway of anti-tumor drug apatinib inhibiting HepG2 cell were studied by NMR metabonomics.The IC₅₀ value of apatinib was firstly determined and the appropriate concentration and the preferred sampling points were determined.Then NMR data collection was performed on cell culture fluids at different time points.The chemometrics methods including principal component analysis were used to analyze the data.The partial least squares and orthogonal partial least squares discriminant analysis models were constructed to identify differential metabolites between the experimental group and the control group.The differential metabolites were verified by one-way ANOVA.The metabolic pathway analysis of 12differential metabolites revealed that the main metabolic pathways of apatinib on HepG2 cells were pyruvate metabolic pathway,alanine,aspartic acid and glutamate metabolic pathway.In the second part,preliminary studies were carried out on the antitumor activities of 12 new coumarin derivatives.Cytotoxicity tests were performed on 12 new coumarin derivatives with using HepG2 cells,and the IC₅₀ concentrations of 12 compounds were obtained,ranging from 60-100?M.HepG2 cells were cultured according to IC₅₀concentrations.The cell culture medium collected at 48 hours was collected for NMR data.The data were normalized and the principal component analysis was performed.The scores of the experimental group and the control group on the first principal component ranged from 60%to 80%,indicating that the composition of the cell culture medium between the experimental and the control groups were very different.According to the order of importance of variable weights and the correlation coefficients of the variables related to the first prediction principal component in the orthogonal partial least squares discriminant analysis model,the load map of orthogonal partial least squares discriminant analysis model was constructed.Differential metabolites between the components of the culture medium in the experimental group of the coumarin derivatives and the negative control group were identified.This study should provide valuable methodological reference for the development of new drugs and the evaluation of the activity of small molecule compounds.