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Diabetes mellitus caused 3.4 million deaths in 2024. Although α-glucosidase and DPP-IV inhibitors are utilised therapeutically, synthetic drugs cause side effects. Durio kutejensis was examined for antihyperglycemic potential. This study assessed the antihyperglycemic activity of the leaves and bark of D. kutejensis from four Kalimantan provinces and identified active compounds through untargeted metabolomic analysis. Ethanol extracts were tested for in vitro α-glucosidase and DPP-IV inhibitory activities, confirmed through in vivo, and followed by untargeted metabolomic via UHPLC-Q-Orbitrap-HRMS. Bark extracts suppressed α-glucosidase activity with IC₅₀ values ranging from 1.65 to 15.10 µg/mL. A total of 44 metabolites were putatively identified; 12 were linked to α-glucosidase inhibition. In vivo, East Kalimantan bark extract (KT2) significantly improved glycemic control (p < 0.05), with a reduction in blood glucose of 24 mg/dL. These findings reinforce the therapeutic value of D. kutejensis bark as an antihyperglycemic agent.