Green Extraction of Phenolic Compounds From Pistacia lentiscus Leaves Coupled With LC‐MS Characterization and Structure‐Based Enzyme Inhibition Analysis

Abstract

Acetylcholinesterase (AChE) and urease are critical enzymes involved in various pathological conditions associated with cellular and tissue damage. Natural products are increasingly explored as safer alternatives to synthetic inhibitors. This study evaluates the phytochemical composition and enzyme inhibitory potential of Pistacia lentiscus leaves using a green extraction approach. Bioactive compounds were extracted via lactic acid‐assisted ultrasound, and the crude extract was characterized for total phenolics, flavonoids, LC‐MS profile, and in vitro inhibition of acetylcholinesterase and urease. Molecular docking was performed to investigate ligand–enzyme interactions. The extract contained 142.13 ± 0.06 mg GAE/g DW of polyphenols and 16.17 ± 0.02 mg QE/g DW of flavonoids. LC‐MS analysis identified phenolic acids, phenolic glycosides, coumarins, and multiple flavonoid subclasses, including flavonols, flavanols, flavanones, flavones, and flavonolignans. The extract exhibited significant inhibition of both enzymes. Docking studies revealed key interactions, such as hydrogen bonds, π‐alkyl, and π‐sigma, within the active sites, with hesperidin and verbascoside showing the strongest binding affinities (acetylcholinesterase: –10.1 and –10.2 kcal/mol; urease: –9.6 and –9.2 kcal/mol, respectively). These results demonstrate that ultrasound‐assisted lactic acid extraction efficiently isolates phenolic compounds from P. lentiscus, and the identified metabolites may serve as promising templates for the development of potent enzyme inhibitors.