Benzimidazolones have shown biological activities, including antihyperglycemic and hypoglycemic, by inhibiting or activating of α-glu and GK. The aim of this study is the rational design of compounds usingin silicoassays to delimitate the selection of structures to synthesize and thein vitroevaluation of benzimidazolone derivatives in blood glucose control. A docking of 23 benzimidazolone derivatives was performed; selecting the compounds with betterin silicoprofiles to synthesize by microwave-irradiation/conventional heat and evaluate in enzymaticin vitroevaluation. Compounds 2k, 2m, 2r, and 2s presented the bestin silicoprofiles, showing good affinity energy (−10.9 to −8.6 kcal mol−1) and binding with catalytic-amino acids. They were synthesized at 70 °C and 24 h using DMF as the solvent and potassium carbonate (yield: 22–38%). The results with α-glu showed moderate inhibition of 2k (14 ± 1.23–29 ± 0.45), 2m (12 ± 2.21–36 ± 0.30), 2r (7 ± 2.21–13 ± 1.34), and 2s (11 ± 0.74–35 ± 2.95) at evaluated concentrations (0.1 to 100 μg mL−1). The GK activation assay showed an enzymatic activity increase; compound 2k increased 1.31 and 2.83 more than normal activity, 2m (2.13-fold), 2s (2.86 and 3.74-fold) at 100 and 200 μg mL−1respectively. The present study showed that the 2s derivative presents moderate potential as an α-glu inhibitor and a good activator potential of GK, suggesting that this compound is a good candidate for blood glucose control through antihyperglycemic and hypoglycemic mechanisms.