Objective: Diabetes mellitus (DM) causes hyperglycemia, which is one of the most common diseases in the world. One of the strategies for the treatment of diabetes is maintaining postprandial glucose level through inhibition of dipeptidyl peptidase IV (DPP-IV) and α-glucosidase enzymes. The aim of this study was to determine in vitro antidiabetic potential of Ferula assa-foetida via DPP-IV and α-glucosidase inhibitory activities.Methods: F. assa-foetida seeds were extracted in methanol, ethanol, ethanol-methanol, and water. Inhibitory activity on DPP-IV and α-glucosidase was performed in vitro and measured spectrophotometrically at λ=405 nm. Results:The result showed that the F. assa-foetida seed extract is effective against both enzymes. All fractions had DPP-IV inhibitory activity, but the ethanolic fraction had the highest inhibitory activity on DPP-IV enzyme and significantly decreased DPP-IV activity (24.5%). With respect to α-glucosidase inhibitory activity, the aqueous extract has the highest inhibitory activity (28%). Conclusion:According to the results of this study, F. assa-foetida contains DPP-IV and α-glucosidase inhibitors and could be a potential source for the discovery of active constituents as α-glucosidase and DPP-IV inhibitors to treat Type 2 DM.