Objective Network pharmacology,Molecular docking and Label-free DIA quantitative phosphoproteomics were used to reveal the potential mechanism of Peiminine against colon cancer.Methods 1The target of peiminine was obtained by SwissTargetPrediction,TargetNet and pharmmapper,and the target of colon cancer was obtained by DisGeNET,GeneCards and OMIM.Then the intersection target of Peiminine and Colon cancer was obtained by Venny2.1.0 online platform.Then,String database and Cytoscape3.8.2 software were used to map the PPI network of intersection targets,and the main targets of Peiminine against Colon cancer were obtained.GO analysis and KEGG pathway analysis were carried out through David database and Weisenxin visual cloud platform.② MOE(molecular operating environment)software was used to perform molecular docking of peiminine and the main target.③ Label-free DIA quantitative phosphoproteomics was used to detect and analyze the biological function of DT group(DT1-DT3)treated with Peiminine and control group(NC1-NC3).Results ① There were 275 intersection targets between peiminine and colon cancer.Molecular docking showed that peiminine could stably dock and interact with the protein structures of AKT1,EGFR,HSP90AA1 and SRC:Peiminine interacted with the amino acid residues of AKT1 mainly through hydrogen bonding.Peiminine interacted with amino acid residues of EGFR,HSP90AA1 and SRC mainly through ionic bond and hydrogen bonding.② Phosphoproteomics analysis showed that:Compared with the NC group,880 phosphorylated modification sites were significantly up-regulated in the DT group(including S124 and S126 sites of AKT1 and T648 and S643 sites of EGFR),and 425 phosphorylated modification sites were significantly down-regulated in the DT group(including T317 sites of HSP90AA1).③ Comparing the results of network pharmacology and phosphoproteomics analysis,it was found that:The main targets of Peiminine against Colon cancer are AKT1,EGFR and HSP90AA1.It promotes apoptosis of colon cancer cells by regulating 17 pathways including AMPK signaling pathway,mTOR signaling pathway and Choline metabolism in cancer.Conclusion This study revealed the potential mechanism of peiminine in the treatment of colon cancer with multiple targets and multiple pathways,and provided a certain direction and reference for subsequent research.

Objective Network pharmacology,Molecular docking and Label-free DIA quantitative phosphoproteomics were used to reveal the potential mechanism of Peiminine against colon cancer.Methods 1The target of peiminine was obtained by SwissTargetPrediction,TargetNet and pharmmapper,and the target of colon cancer was obtained by DisGeNET,GeneCards and OMIM.Then the intersection target of Peiminine and Colon cancer was obtained by Venny2.1.0 online platform.Then,String database and Cytoscape3.8.2 software were used to map the PPI network of intersection targets,and the main targets of Peiminine against Colon cancer were obtained.GO analysis and KEGG pathway analysis were carried out through David database and Weisenxin visual cloud platform.② MOE(molecular operating environment)software was used to perform molecular docking of peiminine and the main target.③ Label-free DIA quantitative phosphoproteomics was used to detect and analyze the biological function of DT group(DT1-DT3)treated with Peiminine and control group(NC1-NC3).Results ① There were 275 intersection targets between peiminine and colon cancer.Molecular docking showed that peiminine could stably dock and interact with the protein structures of AKT1,EGFR,HSP90AA1 and SRC:Peiminine interacted with the amino acid residues of AKT1 mainly through hydrogen bonding.Peiminine interacted with amino acid residues of EGFR,HSP90AA1 and SRC mainly through ionic bond and hydrogen bonding.② Phosphoproteomics analysis showed that:Compared with the NC group,880 phosphorylated modification sites were significantly up-regulated in the DT group(including S124 and S126 sites of AKT1 and T648 and S643 sites of EGFR),and 425 phosphorylated modification sites were significantly down-regulated in the DT group(including T317 sites of HSP90AA1).③ Comparing the results of network pharmacology and phosphoproteomics analysis,it was found that:The main targets of Peiminine against Colon cancer are AKT1,EGFR and HSP90AA1.It promotes apoptosis of colon cancer cells by regulating 17 pathways including AMPK signaling pathway,mTOR signaling pathway and Choline metabolism in cancer.Conclusion This study revealed the potential mechanism of peiminine in the treatment of colon cancer with multiple targets and multiple pathways,and provided a certain direction and reference for subsequent research.