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

Research on plants like Thymelaea hirsuta (L.) Endl. is increasingly crucial for identifying alternative therapies for chronic diseases. This study investigated the n-butanol leaf fraction (NBF) of T. hirsuta for its antioxidant, anti-diabetic, and anti-inflammatory properties through in vitro experiments, including 2,2-diphenyl-1-picrylhydrazyl (DPPH•), 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), Ferric Reducing Antioxidant Power (FRAP), Ferrous Ion Chelating (FIC), α-amylase inhibition, and anti-inflammatory assays. In silico 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). In silico 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 T. hirsuta as a potential source of bioactive compounds for future anti-inflammatory and anti-diabetic pharmaceutical development.