The essential oil (EO) of Origanum minutiflorum O. Schwarz et P. H. Davis was obtained by the hydrodistillation method. The analysis of the EO was conducted using Gas Chromatography (GC) and Gas Chromatography/Mass Spectrometry (GC/MS). The yield of the EO was determined to be 3.9%. Fifty-six compounds were identified, constituting 97.8% of the EO. The EO was rich in carvacrol at a rate of 83.3%, and the other major compounds were p-cymene (3.0%), β-caryophyllene (1.3%), trans-sabinene hydrate (1.1%), γ-terpinene (1.1%), borneol (1.1%), and terpinene-4-ol (1.0%). While the EO (100 μg/mL) demonstrated the highest inhibition on cyclooxygenase-1 (COX-1) with 55.26%, the inhibitory activities on the other enzymes were as follows: cyclooxygenase-2 (COX-2); 33.10%, and matrix metalloproteinase-9 (MMP-9); 12.87%. The EO had no inhibition on lipoxygenase (LOX). In this research, inhibitory activity of the EO of O. minutiflorum on COXs and MMP-9 enzymes was reported for the first time.