Integrated Phylogenetic, Phytochemical, and In Silico Evaluation of Equisetum diffusum D. Don. Ethanolic Extract for Bioactivity
Nasir Assad, Muhammad Naeem‐ul‐Hassan, Muhammad Fayyaz ur Rehman, Yasir Assad, Hansa Gul, Marzia Batool Laila, Muhammad Nauman Khan, Zahida Nasreen, Muhammad Adnan, Alevcan Kaplan, Sezai ErcişliABSTRACT
This study investigated the antibacterial and antioxidant potential of Equisetum diffusum D. Don. using in vitro assays, molecular docking, GC‐MS, HPLC, phytochemical screening, ADME, toxicity, and phylogenetic analyses. Phytochemical screening confirmed the presence of alkaloids, flavonoids, phenols, saponins, and fatty acids in the extracts. GC‐MS and HPLC analyses revealed the presence of bioactive compounds, including 7,9‐Di‐tert‐butyl‐1‐oxaspiro(4,5)deca‐6,9‐diene‐2,8‐dione, oleic acid, and bis (2‐ethylhexyl) phthalate. The oxaspiro compound exhibited the strongest docking affinity with antimicrobial (PDB: 1R4U, −7.4 kcal/mol) and antioxidant (PDB: 1AJ6, −7.3 kcal/mol) targets. In vitro antibacterial assays confirmed the inhibition of Gram‐positive and Gram‐negative strains, whereas DPPH, FRAP, and TFC assays validated strong antioxidant activity. ADME analysis indicated favorable pharmacokinetics for most compounds, although bis (2‐ethylhexyl) phthalate showed toxicity risks. Phylogenetic analysis revealed conserved protein targets across Equisetum species. These findings highlight E. diffusum as a promising source of natural antioxidants and antimicrobial agents, supporting further in vivo and formulation studies for pharmaceutical development.