IntroductionMedicinal plants are considered as rich resources of secondary metabolites and have been used for treating diseases. Pomegranate (Punica granatum) is considered “a pharmacy unto itself”. The current study aimed to evaluate the effect of ripening and variety factors on the antioxidant activity of pomegranate fruits and their capacity to inhibit three enzymes involved in degenerative diseases. Material and methodsThe fruits were collected from 4 Tunisian pomegranate varieties (Espagnoule, Rafrafi, Zaghwani and Zehri) during 3 maturity stages. Phytochemical content, the antioxidant activity by DPPH and ABTS assays and acetylcholinesterase, butyrylcholinesterase and 15-lipoxygenase inhibition activities were carried out. ResultsThe unripe fruits were found to be the richest in terms of total phenolic content. Total flavonoids, tannins, anthocyanins and acetylcholinesterase, butyrylcholinesterase and 15-lipoxygenase inhibition activities varied greatly during ripening process and within varieties. It was found that unripe fruits of Zehri (IC50 = 2.2 µg/ mL by ABTS assay) and Rafrafi (IC50 = 73.4 µg/ mL by DPPH assay) varieties exhibited the highest antioxidant capacities. The strongest extract against acetylcholinesterase and butyrylcholinesterase was found to be the unripe fruit extract of Espagnoule variety with IC50 values of 86.7 and 58.0 µg/ mL, respectively. The ethanolic extract from the unripe fruit of Zehri pomegranate variety inhibited moderately 15- lipoxygenase activity with an IC50 value of 63.0 µg/ mL. ConclusionThe current study supported the popular use of pomegranate fruits, and highlighted also the potential effect of variety and ripening factors on their chemical composition and in vitro biological activities.