•Novel urea derivatives as dual-acting agonists of GK and PPARγ were synthesized.•Several agonists exhibited high enzyme activity and moderate hypoglycemic efficacy.•The discovery may provide an effective approaching for treating T2DM.Motivated by the discovery of a potential ligand that activates both glucokinase (GK) and perioxisome proliferator-activated receptor-γ (PPARγ), this work presents the rational design and synthesis of a series of novel urea derivatives as potent dual-target ligands of GK and PPARγ. The derivatives obtained, particularly compounds 14j, 14m, 15g, 15j, and 15s, showed relatively high enzyme activity and moderate blood glucose-lowering efficacy in normal ICR mice (GK activation fold >1.7, PPARγ activation percentage >38.8%, relative to rosiglitazone). The discovery of a dual-acting agent may provide an effective approach for treating type 2 diabetes mellitus.A novel series of urea derivatives as potent dual-acting agonists of GK and PPARγ were designed and synthesized. The binding model was predicted by molecular docking simulation.