The Research Of Drug Screening And Mechanism Of Drug Action Using Microfluidic And Metabolomics Technolgy

The early prediction of the metabolic fate and their toxicity is the essential and important part of the drug development process. Also, exploring the mechanism of antitumor activity of drugs can be helpful for the design and development of the new drug. In our work, we aimed to develop an integrated microfluidic device for simultaneously characterization of drug metaoblites and cytotoxicity assay. And using metabolomics to explain the mechanism of antitumor activity of flavopiridol.In this work, an integrated microfluidic device was developed for simultaneously characterization of drug metaoblites and cytotoxicity assay. The multi-layer device was composed the functional units of drug metabolism and cytotoxicity assays including metabolite generation, sample pretreatment, online detection with an ESI-Q-TOF MS and incubation with cultured cells to evaluate metabolism induced cytotoxicity. By aligning the PC membrane with a microwell on the PDMS layer, HLMs could be easily immobilized inside the microwell without any damage. To verify the feasibility of our established platform, the metabolism of FLAP and the cytotoxicity of metabolites on MCF7, MCF10A, HepG2and QSG7701cells were studied. The total analysis time for one product was about30min and only3μL reaction system was required for each experiment. The results demonstrated that this platform is robust for low levels of compounds and shows potential for high-throughput drug screening.FLAP has previously been shown to produce a block in cell cycle progression at either G1or G2. But the mechanism underlying the development of the G1or G2block has not been defined. Here, we presented a global analysis of metabolic changes in MCF7cells treated with FLAP to produce a block in cell cycle progression at either G1or G2, and we will explain the regulation mechanism of FLAP to MCF7cell lines through small molecules. Compared with the control group,82distinct metabolites were found and their structures will be identified through MS/MS spectrum in future. And then, we will find out the pathway that FLAP regulated MCF7cell lines.