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Xinyi Zhang,1 Fengxi Li,2 Rong Li,1 Nan Zhao,2 Dianfeng Liu,1 Yuelin Xu,2 Lei Wang,2 Dongxu Wang,1 Ruihong Zhao3 1Laboratory Animal Center, College of Animal Science, Jilin University, Changchun, 130062, People’s Republic of China; 2Key Laboratory of Molecular Enzymology and Engineering of Ministry of Education, School of Life Sciences, Jilin University, Changchun, 130023, People’s Republic of China; 3Department of Gastroenterology Endoscopy Center, The First Hospital of Jilin University, Changchun, 130021, People’s Republic of ChinaCorrespondence: Dongxu Wang, Laboratory Animal Center, College of Animal Science, Jilin University, Changchun, 130062, People’s Republic of China, Tel +86-18743089730 ; +86-0431-87836570, Email wang_dong_xu@jlu.edu.cn Ruihong Zhao, Department of Gastroenterology Endoscopy Center, The First Hospital of Jilin University, Changchun, 130062, People’s Republic of China, Tel +86-18844115258 ; +86-0431-87836578, Email ruihongzhao8@jlu.edu.cnPurpose: Heterocyclic compounds are organic compounds with heterocyclic structures, which are common in drug molecules. They include pyrazines with diverse functions, including anti-cancer, antimicrobial, antidiabetic, and anticholinergic activities. In this study a new small molecular compound B7 based on tetrazolium substituted pyrazine was synthesized and its effect on the progression of colorectal cancer (CRC) and its potential mechanism were investigated.Methods: We synthesized a series of tetrazolium-substituted pyrazine compounds by chemoenzymatic method. NCM460 (Human), HCT116 (Human), SW480 (Human) cell lines were selected to analyse the inhibitory effect of B7 on CRC by CCK-8, apoptosis, cell migration and invasion, qPCR, Western blotting, molecular docking, immunofluorescence. Moreover, a CRC xenograft model of mice was used to analyzed the role of B7 in vivo.Results: Among these compounds, 3-methyl-5je-6-bis (1H-tetrazole-5-yl) pyrazine-2-carboxylic acid (B7) inhibited CRC cell prolifer