Phenolic profile, <i>in vitro</i> bioactivities, and <i>in silico</i> inhibition of α-amylase and trypsin by <i>Thymelaea hirsuta</i> (L.) Endl.

Research on plants like <i>Thymelaea hirsuta</i> (L.) Endl. is increasingly crucial for identifying alternative therapies for chronic diseases. This study investigated the <i>n</i>-butanol leaf fraction (NBF) of <i>T. hirsuta</i> for its antioxidant, anti-diabetic, and anti-inflammatory properties through <i>in vitro</i> experiments, including 2,2-diphenyl-1-picrylhydrazyl (DPPH^•), 2,2'-azino-<i>bis</i>(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS^•+), Ferric Reducing Antioxidant Power (FRAP), Ferrous Ion Chelating (FIC), α-amylase inhibition, and anti-inflammatory assays. <i>In silico</i> study was also conducted against α-amylase and trypsin enzymes. Eighteen (18) compounds, including various phenolic acids and flavonoids, were identified by HPLC-PDA-MS/MS. The NBF demonstrated potent antioxidant activity, exhibiting 99.60% inhibition against ABTS^•+, 92.85% against DPPH^•, and 88.05% for FIC. It also showed significant α-amylase inhibition (85.40% at 5 mg/mL; IC₅₀: 0.37 ± 0.08 mg/mL). Furthermore, the NBF displayed notable anti-inflammatory activities, with 95.37% inhibition of protein denaturation (IC₅₀: 1.83 ± 0.06 mg/mL), 79.14% inhibition of heat-induced haemolysis (IC₅₀: 0.026 mg/mL), and 77.72% inhibition of proteinase (IC₅₀: 0.81 ± 0.95 mg/mL). <i>In silico</i> molecular docking studies against α-amylase and trypsin enzymes revealed feruloyl apigenin as a promising dual inhibitor. This novel compound exhibited a binding affinity comparable to acarbose and diclofenac sodium. These findings highlight the NBF of <i>T. hirsuta</i> as a potential source of bioactive compounds for future anti-inflammatory and anti-diabetic pharmaceutical development.