Objective: This research aimed to use the Wedelia trilobata leaves in treating diabetes and hypertension in vitro through the inhibitory activities of the involved enzymes such as α-glucosidase, α-amylase, dipeptidyl peptidase IV (DPP-IV) and angiotensin-converting enzyme (ACE). The powdered medicinal herb was extracted by percolation with ethanol 96%, was the total ethanol extract (TP). The amount of the ethanol extract (TP) was dissolved in distilled water and partitioned between n-hexane, chloroform, and n-butanol sequentially to separate the components based on the polarity of each solvent. The water solution after partitioning with n-butanol was the water subfraction. After all, these subfraction extracts were n-hexane (HE), chloroform (CF), n-butanol (BU) and water (WA). All these extracts from the Wedelia trilobata leaves investigated the inhibitory activities of the involved enzymes such as α-glucosidase, α-amylase, dipeptidyl peptidase IV (DPP-IV). Research object and methods: The leaves of Wedelia trilobata were extracted with ethanol, and the subfraction extracts obtained were with n-hexane, chloroform, n-butanol, and water. Subsequently, all extracts were tested for enzyme inhibitory activity using optical densitometry, and the IC₅₀ values were determined to evaluate the in vitro inhibitory activity of α-glucosidase, α-amylase, DPP-IV (for diabetes), and ACE (for hypertension). Results: The highest α-glucosidase, α-amylase, angiotensin-converting enzyme (ACE), and DPP-IV inhibitory activities were in CF extract with IC50 of 170.08 μg/mL, 136 μg/mL, 60.67 μg/mL, 108.99 μg/mL, respectively. Conclusion: The leaves of the Wedelia trilobata are a potential source of medicinal herbs in treating diabetes and hypertension.