Chemical Proteomice-Based Targets Investigation Of Bioactive Natural Products

At present,more than half of the small molecule new drugs in the world are directly or indirectly derived from natural products(NPs)and their derivatives.Using active natural products as lead compounds is an important way to develop new drugs.The research and development of new drugs is an extremely time-consuming and costly process.In the initial stage of drug research and development,identifying the target and mechanism of the lead compound can not only understand the causes of its toxicity and side effects,but also conduct targeted structural modification.Although the number of bioactive small molecules is increasing,target identification is still a major problem in medicine and biology,which hinders the wide use in biology and medicine.Therefore,effective research strategies for active natural product targets are very important,which can advance the process of drug development,shorten time and improve efficiency.Among many emerging target research strategies,chemical proteomics is widely used for its unique advantages.It is a powerful tool for characterizing drug-target interactions.At the same time,it can identify adverse reactions with proteins other than the targets,help predict side effects,and help optimize drug candidate molecules.The core technology of chemical proteomics is the design and synthesis of small molecule probes.Small molecule probes must have similar biological activity and selectivity as lead compounds.This project uses chemical proteomics to identify targets of our active natural product.We have selected natural products that exhibited good biological activity in the preliminary in vitro activity screening.Based on the preliminary structure-activity studies of the lead compounds,biotinylated the active natural products to form a small-molecule probe.The cell was incubated with a small molecular probe to make it specifically bind to proteins.The biotin-streptavidin(BSA)system is used to enrich and purify proteins.Two chemical proteomics strategies are used: gel-based affinity proteomics combines MS-based affinity proteomics.Finally,the protein samples are processed into peptide samples that can be identified by MS,and then identified by high-resolution MS.After data analysis,the enriched proteins were selected compared with the control groups.The enriched proteins were analyzed by GO and KEGG,and the comprehensive biological functional annotation was obtained.Combining the mass spectrometry data of two chemical proteomics strategies,the most likely protein targets were predicted.We used chemical proteomics to identify the active natural products,and they may function in DNA replication,DNA synthesis,transcription and translation of proteins and mitotic cell cycle pathway.At the same time,five protein targets were predicted.This research provides evidences that chemical proteomics is an effective way to identify the targets of active natural products and provides reference for the identification of other active natural product targets.