Optimizing biofilm inhibitors: Balancing activity and toxicity in 2N-aminated 5-aryl-2-aminoimidazoles

To evaluate the effect of amination on biofilm inhibition against Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus, representative compounds of two previously described 5-aryl-2-aminoimidazole (5-Ar-2-AI) classes were aminated by installing an amino group at the end of the substituted n-alkyl chain. Amination led to an improvement in activity for one of the two classes, the 2N-substituted 5-Ar-2-AI class. Based on these findings, a more extensive library of 2N-substituted-aminated 5-Ar-2-AIs was synthesized having different n-alkyl and halogen substitutions on the 2N-position and the 4(5)-phenyl ring, respectively. Compounds were evaluated for their biofilm inhibitory activity against E. coli, P. aeruginosa, S. aureus, Staphylococcus epidermidis and MRSA. Additionally, their toxicity was tested on eight continuous cell lines, peripheral blood mononuclear cells and Caenorhabditis elegans, along with their genotoxicity on Capan-1. Halogenation and elongation of the n-alkyl substituent showed a positive effect on biofilm inhibitory activity, but also increased toxicity. Compromising between activity and toxicity, a non-halogenated 2N-substituted-aminated 5-Ar-2-AI compound with an intermediate n-heptyl substitution demonstrated promising broad-spectrum biofilm inhibition, making it a suitable candidate for further research in anti-infectious medical applications. © 2025 Elsevier B.V., All rights reserved.