In-silico molecular docking and in vitro cytotoxic evaluation of Compounds 6, 10, and 14 against MCF-7 cells. Structural representations, docking interactions, and microscopy images at varying concentrations (1000–31.25 µg/mL) are shown. IC₅₀ values were 170.58, 178.04, and 200.84 µg/mL for Compounds 6, 10, and 14, respectively.

Abstract

Breast cancer metastasis remains a critical challenge, necessitating targeted therapies that disrupt key signaling pathways in tumor progression. Integrin-linked kinase (ILK), a key regulator of cell adhesion, migration, and survival via the AKT/NF-κB axis, is a promising target for anti-metastatic drug development. This study reports the synthesis and evaluation of C2-substituted and unsubstituted imidazolium–triazole derivatives as potential ILK modulators. Compounds were synthesized using Cu(I)-catalyzed azide–alkyne cycloaddition and characterized by FTIR, NMR, and mass spectrometry. To explore upstream molecular interactions, docking studies were conducted using the CXCR4 receptor (PDB ID: 3ODU), functionally linked to ILK activation in metastasis. Compound 14 demonstrated high binding affinity (–9.3 kcal/mol), interacting with Trp94, Asp97, and Ile204, and maintained stability during 100 ns molecular dynamics simulations. In vitro cytotoxicity against MCF-7 cells identified Compound 6 as the most potent (IC₅₀ = 170.58 ± 0.3 µg/mL), with Compound 14 also showing notable activity (IC₅₀ = 200.84 ± 0.7 µg/mL). Structure–activity relationship analysis (SAR) revealed that aromatic and hydrophobic groups enhanced efficacy. These results suggest that imidazolium–triazole hybrids are promising modulators of the CXCR4–ILK–AKT/NF-κB axis and hold potential for further development as anti-metastatic agents in breast cancer therapy.