The rising threat of antimicrobial resistance demands the development of novel therapeutic agents with broad-spectrum efficacy. In this study, a series of thiadiazolyl-pyridine derivatives (5a–5h) was synthesized using a one-pot multicomponent reaction involving acetylthiadiazole, substituted aromatic aldehydes, malononitrile, and ammonium acetate in refluxing acetic acid. This green and efficient synthetic route offered high yields and reduced environmental impact. The structures of the synthesized compounds were confirmed by ¹H NMR, LC-MS, and elemental analysis. The antimicrobial potential of the compounds was evaluated against four bacterial strains (Staphylococcus aureus, Bacillus anthracis, Pseudomonas aeruginosa, Escherichia coli) and two fungal strains (Candida albicans, Aspergillus niger) using the cup plate method. Among the tested compounds, 5d demonstrated the highest antibacterial activity, with inhibition zones comparable to the standard drug streptomycin. Compounds 5c and 5e also exhibited significant antibacterial and antifungal activity. To assess drug-likeness and pharmacokinetic profiles, SwissADME analysis was conducted. All compounds adhered to Lipinski’s Rule of Five, indicating favorable physicochemical properties for oral bioavailability. Furthermore, molecular docking studies were performed against microbial protein targets PDB ID: 2EG7 and 5D6P. Compounds 5c, 5e, and 5g exhibited superior binding affinities compared to streptomycin, highlighting their potential as enzyme inhibitors.
