Methanolic Broccoli Extract as a Potential Multimechanistic Neuroprotective Agent Against Scopolamine‐Induced Cognitive Impairment: An Integrated Analytical and In Vivo Study

ABSTRACT

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cholinergic dysfunction, oxidative stress, and neuroinflammation; however, available AD therapies, such as acetylcholinesterase inhibitors (AChEIs) and memantine, can alleviate only clinical symptoms, which leads to the need for multitarget strategies. In this study, we aimed to perform chemical characterization of a Brassica oleracea var. italica methanolic extract to explore the mechanism underlying its biologically active properties (MEBO), and to assess its neuroprotective potential using combined in silico and in vivo methodologies. Profiling by LC–QTOF–MS revealed that many biochemicals, such as phenolic compounds and nitrogen‐containing secondary metabolites, are present. SwissADME, ProTox‐II, and admetSAR were utilized to assess the drug‐likeness and ADMET properties of the identified compounds, which suggested several constituents with promising pharmacokinetic and safety profiles. Molecular docking against acetylcholinesterase (AChE; PDB ID: 4EY7) confirmed that they bind strongly, with dehydropipernonaline, melosatin B, and ceceline utilizing interactions similar to those of donepezil. In vivo testing via a scopolamine‐induced zebrafish model revealed that MEBO (25–100 mg/L) markedly enhanced cognitive and behavioral function in several paradigms, such as light–dark, novel tank, T‐maze, and novel object recognition (NOR) tests. Biochemical analyses also revealed decreased lipid peroxidation, restoration of antioxidant defense systems, and inhibition of AChE activity. Intermediate toxicity results confirmed the absence of mortality at 100 mg/L; thus, the LD ₅₀ concentration was greater than this concentration. Together, these results indicate that MEBO may have substantial neuroprotective effects, possibly through cholinergic modulation and antioxidant mechanisms, suggesting that this multifunctional product holds promise as a new candidate for AD‐related cognitive impairment. It should be emphasized that these findings were obtained in an acute scopolamine‐induced model of cholinergic dysfunction; accordingly, the present data support a cognitive‐enhancing and neuroprotective potential rather than disease‐modifying efficacy against the amyloid‐ β ‐ and tau‐driven pathology of clinical AD, and warrant confirmation in chronic and genetic models.