Network Pharmacology And Metabolomics Elucidate The Underlying Mechanisms Of The Anti-proliferation Effect Of Bufonis Venenum In Colorectal Cancer

Bufonis Venenum is a traditional Chinese medicine made from the dried secretion of Bufo melanostictus Schneider.Its extract has shown broad anti-tumor effects in clinical practice.Colorectal cancer is a common tumor caused by mutations in colon epithelial cells,with a mortality rate second only to lung cancer worldwide.Previous studies have suggested that Bufonis Venenum has a good therapeutic effect on colorectal cancer.Bufonis Venenum is composed of various chemical compounds such as alkaloids,serotonin,polypeptide,and lactone.Thus its anti-cancer effect is attributed to the synergistic action of multiple effective compounds,which increases the difficulty of studying the anti-cancer effect of Bufonis Venenum.Therefore,there has not been a comprehensive study to elucidate the treatment mechanism of Bufonis Venenum extract on colorectal cancer cell proliferation inhibition.In this study,network pharmacology,metabolomics,and molecular docking techniques were used to explore the mechanisms of Bufonis Venenum’s inhibition of colorectal cancer cell proliferation.Firstly,a "compound-disease’s target" topological network for Bufonis Venenum’s effective compounds in treating colorectal cancer was constructed by using the Chinese medicine resources database,compound target prediction database,and disease target database,and a "protein-protein interaction" network of potential therapeutic targets was established based on this network.Important potential targets were obtained by analyzing the network,and bulk molecular docking was performed with Bufonis Venenum’s effective compounds identified by liquid chromatography-mass spectrometry.The results showed that Bufonis Venenum mainly affects colorectal cancer cell proliferation and cell cycle,with CDK1,TOP2 A,and MTOR as important potential targets.Meanwhile,based on metabolomics analysis of different concentrations of Bufonis Venenum extract-treated groups,it was found that Bufonis Venenum extract inhibits the synthesis of nucleotide triphosphate from de novo and salvage pathways,mainly affecting the metabolic enzymes related to purine and pyrimidine metabolism in colorectal cancer cells,such as NDK,GOT1,APRT,TYMK,and so on,thereby affecting the synthesis of DNA in colorectal cancer cells and causing cell cycle arrest.Finally,the experiments in vitro further confirmed the inhibitory effect of Bufonis Venenum extract on colorectal cancer cell proliferation and its ability to arrest cell cycle in S and G2/M phases.The significance of this study lies in exploring the mechanism of Bufonis Venenum’s inhibition of colorectal cancer proliferation,which further supplements the relevant theoretical basis for the clinical use of Bufonis Venenum in treating colorectal cancer.At the same time,this study successfully predicted the mechanism of Bufonis Venenum’s inhibition mechanism of colorectal cancer proliferation by combining network pharmacology,bulk molecular docking,and multi-dose metabolomics,providing research ideas for similar traditional Chinese medicines.