A novel series of multifunctional benzimidazoles has been reported as potent inhibitors of α-glucosidase. The procedure relies on the synthesis of 5-amino-1H-benzo[d]imidazole-2-thiol 5 via the multistep reaction through 2-nitroaniline 1, benzene-1,2-diamine 2, 1H-benzo[d]imidazole-2-thiol 3, and 5-nitro-1H-benzo[d]imidazole-2-thiol 4. Further treatment of 5 with aromatic aldehydes 6a-m provided access to the target 5-(arylideneamino)-1H-benzo[d]imidazole-2-thiols 7a-m. The results of the bioactivity assessment revealed all the compounds as excellent inhibitors of the enzyme (IC₅₀ range: 0.64 ± 0.05 μM to 343.10 ± 1.62 μM) than acarbose (873.34 ± 1.21). Among them, 7i was the most active inhibitor (IC₅₀: 0.64 ± 0.05 μM) followed by 7d (IC₅₀: 5.34 ± 0.16 μM), 7f (IC₅₀: 6.46 ± 0.30 μM), 7g (IC₅₀: 8.62 ± 0.19 μM), 7c (IC₅₀: 9.84 ± 0.08 μM), 7m (IC₅₀: 11.09 ± 0.79 μM), 7a (IC₅₀: 11.84 ± 0.26 μM), 7e (IC₅₀: 16.38 ± 0.53 μM), 7j (IC₅₀: 18.65 ± 0.74 μM), 7h (IC₅₀: 20.73 ± 0.59 μM), 7b (IC₅₀: 27.26 ± 0.30 μM), 7k (70.28 ± 1.52 μM) and finally 7l (IC₅₀: 343.10 ± 1.62 μM). Molecular docking revealed important interactions with the enzyme, thereby supporting the experimental findings.

This research reports the synthesis of new benzimidazole-derived N-acylhydrazones (NAH), their characterization using various spectroscopic methods, and in vitro evaluation as potent carbonic anhydrase-II inhibitors. Among the target compounds (9–29), few showed higher inhibition than the standard acetazolamide (IC₅₀: 18.6 ± 0.43 μM), for example, compound 9 (IC₅₀: 13.3 ± 1.25 μM), 10 (IC₅₀: 17.2 ± 1.24 μM), 12 (IC₅₀: 14.6 ± 0.62 μM), and 15 (IC₅₀: 14.5 ± 1.05 μM). Molecular docking was performed on the most active compounds, which revealed their binding interactions with the active site of the enzyme, thus supporting the experimental findings.