The current study aimed to investigate the in vitro and in silico antioxidant and anticholinesterase effects of essential oil (EO) and ethanolic extract (EE) from Ammodaucus leucotrichus Coss. & Dur. fruits. GC-MS analysis of the EO revealed a predominance of perillaldehyde (80%), followed by limonene (13.45%). Antioxidant activity results showed that EO had excellent superoxide radical scavenging activity (IC50 = 20.95 μg/mL) compared to α-tocopherol (IC50 = 31.52 μg/mL), while EE demonstrated good ABTS•+ and DPPH radical scavenging activity with IC50 values of 52.06 and 63.51 μg/mL, respectively, compared to BHT (13.01 and 23.14 μg/mL). In addition, noticeable ferric and cupric reducing powers were observed. EO showed good inhibitory activity against both acetyl and butyrylcholinesterase (AChE and BChE) compared to galantamine (199.81 and 86.09 vs. 8.14 and 36.44 μg/mL, respectively). However, very low activity was demonstrated by the EE (IC50 > 200 μg/mL). Molecular docking analysis predicted high binding affinities of perillaldehyde and limonene with AChE and BChE ranging from −5.8 to −6.2 kcal/mol and an inhibition by binding via the π-interaction with the residues of active sites His447 and His438, respectively. Additionally, only perillaldehyde presented hydrogen bonds with both enzymes. Furthermore, the absorption, distribution, metabolism, elimination, and toxicity (ADMET) prediction studies of perillaldehyde and limonene showed good pharmacokinetic properties without inhibition of cytochromes P450 (CYP), nephrotoxicity, or mutagenicity, as well as good drug likeness evaluated by Lipinski’s Rule and a bioavailability score of 0.55. Thus, A. leucotrichus could be a good resource for pharmaceutical applications.
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