Essential Oil and Hydroethanolic Extract of <i>Clinopodium serpyllifolium</i> subsp. <i>serpyllifolium</i> : Enzyme Inhibition, Antiproliferative, Antimicrobial, Antibiofilm and In Silico Insights

ABSTRACT The pharmacological profile of Clinopodium serpyllifolium subsp. serpyllifolium (CS) remains largely unexplored, deserving to be comprehensively investigated. In this regard, the present study was designed to investigate, for the first time, the phytochemical composition and biological activities of CS hydroethanolic extract (CSE) and essential oil (CSEO). The findings were further supported by in silico analyses. Briefly, the metabolite profiling was performed by LC‐ESI‐MS/MS and GC–MS/MS. Concerning in vitro assays, the inhibitory effects of the plant extracts on α‐amylase, α‐glucosidase and xanthine oxidase were assessed. The extracts were also tested for their anti‐inflammatory, antiproliferative (HepG2, SAOS‐2), antibacterial and antibiofilm activities. Additionally, to support the findings regarding biological activities, density functional theory (DFT), molecular docking, molecular dynamics simulations and free energy landscape analyses were employed for the major compounds identified. The findings of the present study revealed that CSEO exhibited potent α‐amylase, α‐glucosidase and xanthine oxidase inhibition. Importantly, CSEO showed superior activities over standard inhibitors. The selective antiproliferative activity of CSE and CSEO was observed in their treatment on hepatocellular carcinoma (HepG2) and osteosarcoma (SAOS‐2) cells, while the normal human umbilical vein endothelial cells (HUVEC) remained unaffected. Also, strong antibacterial and antibiofilm activities were observed, particularly against Pseudomonas aeruginosa and Staphylococcus aureus . Significantly, in silico analyses further confirmed the stable and favourable binding energies of menthone, pulegone and luteolin with target enzymes. Those findings were consistent with experimental findings. Taking all findings into account, the present study provides the first comprehensive evidence that CS might be considered as a promising natural source or candidate for enzyme inhibitors, antiproliferative, anti‐inflammatory and antimicrobial agents.