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AbstractResistance to antibiotics/antibacterials/antifungals in pathogenic microbes has been developing over the past few decades and has recently become a commonplace public-health peril. Thus, alternative nontoxic potent antibiotic agents are covertly needed to control antibiotic-resistant outbreaks. In an effort to combat the challenges posed by the co-occurrence of multidrug resistance, two terpyridine ligands 4′-(4-N,N′-dimethylaminophenyl)-2,2′:6′,2″-terpyridine (L1) and 4′-(4-tolyl)-2,2′:6′,2″-terpyridine (L2) have been designed, prepared and confirmed their structure by spectral studies. Thereafter, antimicrobial assay was performed against gram positive and negative bacterial strains along with fungal strains. Both compounds L1and L2exhibited remarkable inhibitory activities against bacteria, Escherichia coliand Staphylococcus aureusat MIC values 6.25 and 3.125 µg/ml, respectively. In addition, in silicomolecular docking studies were ascertained with bacterial DNA gyrase and fungal demethylase. Furthermore, both L1and L2could bind Bovine Serum Albumin (BSA) protein and binding interaction has been studied with the help of UV-Visible and fluorescence spectroscopy. While fluorescence of BSA unperturbed in the presence of L2, an addition of L1to the solution of BSA resulted significant quenching. The binding constant calculations at different temperature confirmed that the fluorescence quenching between BSA and L1is predominantly static in nature. The toxicity of L1and L2was checked using Drosophila melanogaster. The toxicity analysis suggest both the dyes are non-cytotoxic in nature.Communicated by Ramaswamy H. Sarma