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PHYTOCHEMICAL CHARACTERIZATION, ANTIOXIDANT POTENTIAL AND MULTIFUNCTIONAL ENZYME INHIBITORY ACTIVITY OF HYSSOPUS OFFICINALIS SUBSP. ARISTATUS: AN IN VITRO AND IN SILICO STUDY

TIJANA MIĆOVIĆ 1,2*, STEVAN SAMARDŽIĆ 3, IVONA VELIČKOVIĆ 4, KSENIJA MILESKI 4, JELENA ĐOROVIĆ JOVANOVIĆ ⁵

¹ Institute for Medicines and Medical Devices of Montenegro, Podgorica, Montenegro
2 Faculty of Medicine, University of Montenegro, Podgorica, Montenegro
3 Department of Pharmacognosy, Faculty of Pharmacy, University of Belgrade, Belgrade, Serbia
4 Institute of Botany and Botanical Garden “Jevremovac”, Faculty of Biology, University of Belgrade, Belgrade, Serbia
5 Institute for Information Technologies, University of Kragujevac, Kragujevac, Serbia

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LC-DAD-ESI-MS analysis of the methanol extract from the flowering aerial parts of Hyssopus officinalis subsp. aristatus (Montenegro) revealed the predominance of hydroxycinnamic acid derivatives, flavonoid glycosides, and phenylethanoid glycosides. The extract exhibited moderate antioxidant activity in ABTS, TRC, and BCB assays, with the strongest effect in ABTS (IC50 = 73.41 µg/mL), compared to L-ascorbic acid and BHT (31.59 and 40.37 µg/mL, respectively). Major phenolics, chlorogenic and rosmarinic acids, showed potent antioxidant activity (IC50 = 24.00 and 12.23 µg/mL in the ABTS assay, respectively). The extract showed stronger inhibition of α-glucosidase (IC50 = 19.02 mg/mL) than α-amylase (26.41 mg/mL). The dominant phenolic acids showed potent α-glucosidase inhibition, comparable to acarbose. This study is the first to report inhibitory activity of this species against arginase (IC50 = 7.47 mg/mL) and elastase (IC50 = 26.24 mg/mL). Molecular docking indicated favorable binding of rosmarinic acid to α-glucosidase (ΔGbind = -7.56 kcal/mol) and arginase (ΔGbind = -7.77 kcal/mol), providing a theoretical rationale for the extract's activity. These findings highlight this plant as a promising source of multifunctional phenolics relevant to oxidative stress and metabolic disorders.