Design, Synthesis and Antiproliferative Activity Studies of Novel 1,2,3-Triazole-Urea Hybrids

En Gao,1,2 Haoying Zhang,1 Yajie Guo,3 Xi Wang,1 Longfei Mao,4 Lizeng Peng,5 Guiqing Xu,1 Xiaojun Yao,2 Shouhu Li,6 Huibao Long,3 Tong Wang,3 Haidong Wu3 1School of Chemistry and Chemical Engineering, Henan Normal University, Xinxiang, 453000, People’s Republic of China; 2State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Macau, People’s Republic of China; 3Department of Emergency, the Eighth Affiliated Hospital, Sun Yat-Sen University, Shenzhen, 518033, People’s Republic of China; 4College of Basic Medicine and Forensic Medicine, Henan University of Science and Technology, Luoyang, 471003, People’s Republic of China; 5Institute of Agro-Food Science and Technology, Shandong Academy of Agricultural Sciences, Jinan, 250100, People’s Republic of China; 6School of Pharmacy, Xinxiang University, Xinxiang, Henan, 453000, People’s Republic of ChinaCorrespondence: En Gao, School of Chemistry and Chemical Engineering, Henan Normal University, Xinxiang, 453000, People’s Republic of China, Email [email protected] Lizeng Peng, Shandong Academy of Agricultural Sciences, 202 Gongye North Road, Licheng District, Jinan City, 250000, People’s Republic of China, Email [email protected]: The development of novel targeted therapies for hepatocellular carcinoma remains a critical need in oncology. This study aimed to design and evaluate a new series of triazole-urea hybrid compounds for their anti-cancer potential.Methods: A series of urea compounds bearing terminal alkynyl groups were synthesized via an assembly and post-modification strategy. Subsequent Click chemistry yielded the novel triazole-urea hybrids ( 3a– 3e, 5a– 5e, 7a– 7e). Their anti-proliferative effects were assessed against multiple human cancer cell lines (lung: H460, H1299, A549, PC-9; liver: Huh-7; breast: MCF-7) and normal liver cells (L02) using the CCK-8 assay. Mechanisms were investigated through apoptosis, autophagy, and DNA damage assays. An acute oral toxicity study was conducted in female KM mice at a dose of 500 mg/kg, with thorough monitoring of body weight, organ coefficients, and histopathology of major organs.Results: Most compounds exhibited potent, concentration- and time-dependent anti-proliferative activity against Huh-7 liver cancer cells, with only marginal effects on other tested cancer lines. Crucially, no cytotoxicity was observed in normal L02 cells. Mechanistic studies revealed that the lead compound induced apoptosis, autophagy, and DNA damage in Huh-7 cells. The in vivo assay demonstrated no drug-related mortality or significant adverse effects on body weight, organ coefficients, or histology at 500 mg/kg over 14 days, indicating a high maximum tolerated dose and an excellent preliminary safety profile.Conclusion: These findings demonstrate the selective anti-liver cancer efficacy and favorable in vivo safety of these triazole-urea hybrids, particularly compound 3c, underscoring their strong potential as promising therapeutic candidates for hepatocellular carcinoma.Keywords: 1,2,3-triazoles, urea, hepatic cancer, anticancer activity