The need for effective targeted agents against hepatocellular carcinoma remains pressing. This investigation focused on designing and examining a new panel of triazole–urea hybrid molecules for potential anticancer activity. A collection of urea derivatives containing terminal alkynes was generated using a combined assembly and modification route. These intermediates were subsequently transformed through Click chemistry to afford novel triazole–urea hybrids (3a–3e, 5a–5e, 7a–7e). Their growth-inhibitory effects were evaluated via the CCK-8 assay against several human tumor cell lines (lung: H460, H1299, A549, PC-9; liver: Huh-7; breast: MCF-7) and normal hepatocytes (L02). Mechanistic analyses included apoptosis induction, autophagy markers, and DNA damage responses. An acute toxicity assessment was carried out in female KM mice at an oral dose of 500 mg/kg, with continuous monitoring of body mass, organ indices, and histological changes in primary organs.
Most synthesized molecules showed marked, dose- and time-dependent suppression of Huh-7 cell proliferation, while only minor activity was detected in other malignant lines. Importantly, none displayed toxicity toward L02 normal liver cells. Mechanistic assays confirmed that the lead candidate triggered apoptotic processes, autophagic responses, and DNA damage in Huh-7 cells. In vivo, no treatment-related deaths or notable alterations in weight, organ coefficients, or tissue histology were observed during the 14-day evaluation at 500 mg/kg, indicating a high tolerated dose and favorable initial safety characteristics. These outcomes highlight the selective anti-hepatoma activity and promising safety of the triazole–urea scaffold, especially compound 3c, supporting its potential for further development against hepatocellular carcinoma.
