In the context of increasing antibiotic resistance, the development of novel small molecules exhibiting selective, narrow-spectrum inhibition of bacterial-specific enzymes or proteins—with high potency and reduced adverse effects—represents a key priority in drug discovery. Accordingly, we established and executed a successful methodology for identifying new hybrid structures, particularly the relatively underexplored [2-(3-R-1H-1,2,4-triazol-5-yl)phenyl]amine derivatives. These scaffolds function as modular platforms, facilitating the attachment of specific functional groups to improve activity against staphylococcal pathogens. The desired compounds were synthesized through streamlined single-reaction protocols, utilizing transformations of suitably substituted 4-hydrazinoquinazolines or 2-aminobenzonitrile starting materials in combination with appropriate carboxylic acid reagents. Putative interactions as DNA gyrase inhibitors were examined via computational docking simulations, supporting their assessment for antistaphylococcal potential. A considerable number of the prepared analogs demonstrated strong inhibitory activity toward Staphylococcus aureus (MIC values: 10.1–62.4 µM). In particular, 5-bromo-2-[3-(furan-3-yl)-1H-1,2,4-triazol-5-yl]aniline and 5-fluoro-2-[3-(thiophen-3-yl)-1H-1,2,4-triazol-5-yl]aniline displayed MICs of 5.2 µM and 6.1 µM, respectively, nearly matching the reference compound ciprofloxacin (MIC: 4.7 µM). Structure-activity relationship (SAR) studies and ADME predictions highlight opportunities for further optimization. The [2-(3-R-1H-1,2,4-triazol-5-yl)phenyl]amine series exhibits notable antimicrobial efficacy and justifies advanced exploration as targeted antistaphylococcal therapeutics. SAR findings emphasize that cycloalkyl or electron-rich heterocyclic groups at the triazole 3-position are critical for activity, whereas methylation of the aniline ring increases potency. Halogen incorporation into the aniline ring produces mixed effects, varying with the triazole 3-substituent. The synthesized [2-(3-R-1H-1,2,4-triazol-5-yl)phenyl]amines show substantial antistaphylococcal effects and warrant in-depth investigation as promising antibacterial candidates.
