Comprehensive Chemical and Biological Assessment of Orobanche racemosa Extracts Using Chromatographic, In Vitro, In Silico, and Cell-Based Techniques

The growing interest in natural product research as a safer and sustainable alternative to synthetic drugs has triggered this study, which evaluates the chemical profile and biological activity of Orobanche racemosa extracts. In the current study, extraction was performed from O. racemosa using the maceration method with several extraction solvents. The extracts were examined for their antioxidant and enzyme-inhibitory activities and their effects on cell viability. In high-performance liquid chromatography–MS/MS analysis, a total of 31 compounds was detected across all studied extracts. The EtOH and EtOH/water extracts consistently exhibited the highest antioxidant capacities in almost all tested assays. Similarly, EtOH and EtOH/water extracts showed vigorous anti-AChE activity, measuring 2.88 and 2.35 mg/GALAE, respectively. In the cell viability assay, both EA and EtOH extracts of O. racemosa demonstrated potent cytotoxicity, significantly reducing viability in normal (HEK293) and cancer cell lines (HepG<inf>2</inf>, SH-SY5Y), indicating high toxicity without selectivity. Molecular docking screened 384 theoretical compound–target combinations and retained 251 unique complexes with docking scores of −7.0 kcal/mol or lower after duplicated entries were removed. Subsequent single-trajectory molecular dynamics analysis of seven representative high-scoring complexes generated hypotheses regarding structural persistence under the simulated conditions; however, these computational predictions require experimental validation.