Ethyl 4-(1H-tetrazol-1-yl)benzoate derivatives as multi-target inhibitors of cancer migration via BTK, BRAF, and FAK modulation: in silico and in vitro studies

Metastasis is the leading cause of cancer-related mortality, driven by complex processes such as cell migration and focal adhesion dynamics, which are regulated by key kinases including BTK, BRAF, and FAK. In this study, we designed, synthesized, and comprehensively evaluated a novel series of ethyl 4-(1H-tetrazol-1-yl)benzoate derivatives (Z1-Z7) as potential multi-target inhibitors of cancer metastasis. In silico analyses, including molecular docking and molecular dynamics simulations (MDs), identified compounds Z3 and Z4 as the most promising candidates, demonstrating strong and stable interactions with BTK, BRAF, and FAK. Density Functional Theory (DFT) calculations further supported these findings, revealing that Z3 and Z4 exhibited the lowest HOMO-LUMO energy gaps, indicating a high potential for charge transfer. Based on these results, Z3 was selected for in vitro validation, where it exhibited notable anticancer activity against A549 (IC₅₀ = 2.522 μg/mL) and SKGT4 (IC₅₀ = 1.576 μg/mL) cancer cell lines, while also displaying cytotoxicity toward normal Vero cells (IC₅₀ = 2.758 μg/mL). Additionally, Z3 significantly inhibited cancer cell migration in wound healing assays, with effective concentrations of 0.5 μg/mL, 2.5 μg/mL, and 5 μg/mL for A549 cells, and 0.5 μg/mL, 1.6 μg/mL, and 3.2 μg/mL for SKGT4 cells. These findings suggest that ethyl 4-(1H-tetrazol-1-yl)benzoate derivatives, particularly Z3 and Z4, hold promise as multi-target therapeutic agents capable of modulating critical pathways involved in cancer migration and metastasis. With further optimization and validation, these compounds may contribute to the development of next-generation targeted therapies that address both tumor growth and the serious challenge of metastasis.