The Biopotential of Bellardia trixago in Replacing Synthetic Compounds for Health-Promoting Applications: Is It a Promising Candidate?

Bellardia trixago (L.) All. (family Orobanchaceae) is a facultative hemiparasitic plant used traditionally to cure many diseases. The present study was designed to evaluate the phenolic constituents, antioxidant, enzyme inhibitory, and cytotoxic properties of the aerial parts of B. trixago. Ethyl acetate (EtOAc), ethanol (EtOH), and 70% EtOH extracts were prepared by maceration, while the aqueous extract was prepared by infusion. Rutin, 4-hydroxy benzoic acid, ferulic acid, p-coumaric acid, hyperoside, delphinidin 3,5 diglucoside, kaempferol, and isoquercitrin were identified in most extracts with variable concentrations, but generally, the EtOH or 70% EtOH accumulated the highest contents. The 70% EtOH extract displayed the best antiradical (DPPH = 13.55 mg TE/g; ATBS = 53.78 mg TE/g) and ion-reducing (CUPRAC = 50.09 mg TE/g; FRAP = 24.73 mg TE/g) properties. The aqueous extract recorded the highest chelating iron power (12.91 mg EDTAE/g) while EtOAc and EtOH exerted the highest total antioxidant activity (2.29 and 2.49 mmol TE/g, p > 0.05). The best cholinesterase (anti-AChE = 2.91 mg GALAE/g; and anti-BchE = 2.62 mg GALAE/g) and α-glucosidase (1.19 mmol ACAE/g) inhibition activity was recorded from the EtOH extract, while that against the α-amylase was obtained from the EtOAc extract (0.69 mmol ACAE/g). The plant was most cytotoxic toward the HT-29 cell line, where the best effect was exerted by the 70% EtOH (IC50 38.42 μg/mL) extract. Furthermore, network pharmacology analysis revealed critical gene targets and pathways associated with the bioactive compounds. Molecular docking studies revealed favorable binding modes and interaction patterns of major compounds with key colon cancer-related proteins, which were further supported by molecular dynamics simulations. As the first investigation into the phenolic composition and pharmacological effects of B. trixago, this study highlights its potential as a promising source of bioactive compounds with therapeutic relevance for oxidative stress-related conditions.