Bioactive Potential and COX-2 Interaction of Ajuga iva (L.) Schreb. Hydroalcoholic Extract: Evidence from Experimental and Computational Studies

Ajuga iva (L.) Schreb. is traditionally used in North African ethnomedicine for the management of inflammation, pain, and fever. The present study aimed to characterize the phytochemical profile of the hydroalcoholic extract of its aerial parts and to evaluate its anti-inflammatory, analgesic, and antipyretic activities using established in vivo models. Preliminary phytochemical screening confirmed the presence of major classes of secondary metabolites, including polyphenols, flavonoids, tannins, and glycosidic compounds. Quantitative assays revealed appreciable levels of total phenolics (26.3 ± 1.2 mg GAE/g extract) and flavonoids (13.5 ± 0.9 mg QE/g extract). In vivo pharmacological evaluation demonstrated significant biological activities, with the highest tested dose (400 mg/kg) producing a marked inhibition of carrageenan-induced paw edema (44.9%), comparable to acetylsalicylic acid. At the same dose, the extract showed pronounced analgesic activity in the acetic acid-induced writhing test, with an inhibition rate of 64.2%, and a significant antipyretic effect in the brewer’s yeast-induced fever model, as evidenced by a reduction in rectal temperature. In parallel, molecular docking was employed as an exploratory, hypothesis-generating in silico approach to investigate potential interactions between selected phenolic constituents identified in A. iva and cyclooxygenase-2 (COX-2). Several compounds, including rosmarinic acid, rutin, and apigenin-7-O-glucoside, displayed favorable predicted binding affinities and interactions with key residues of the COX-2 active site. It should be emphasized that molecular docking was used solely as a hypothesis-generating in silico tool and does not constitute direct biochemical evidence of COX-2 inhibition. Overall, these findings indicate that the hydroalcoholic extract of Ajuga iva exhibits notable anti-inflammatory, analgesic, and antipyretic activities in vivo. The in silico docking results provide supportive, predictive molecular insights that may help rationalize the observed bioactivities and encourage further biochemical and mechanistic investigations into this traditionally used medicinal plant.