© 2015, Turkish Pharmacists Association. All rights reserved.In the current work, new benzimidazole-based acetamide derivatives (2a-u) were synthesized and screened for their in vitro antimicrobial activity. Among these derivatives, compounds 2b-2g were found to be the most promising antibacterial agents against Pseudomonas aeruginosa. These compounds and streptomycin exhibited the same level of antibacterial activity with a MIC value of 125 μg/mL. Compounds 2p, 2s, 2t and 2u were the most potent antifungal derivatives against Candida krusei with a MIC value of 125 μg/mL when compared with ketoconazole (MIC= 62.5 μg/mL). Compounds 2s and 2u also exhibited the highest inhibitory activity against Fusarium solani with a MIC value of 125 μg/mL, whereas ketoconazole showed its antifungal activity with a MIC value of 62.5 μg/mL.