Clitoria mariana L., a medicinal plant widely distributed in Asia, has long been used in folk medicine. This study investigates the biomedical potential of its methanolic flower extract. Gas chromatography–mass spectrometry (GC–MS) and phytochemical analyses revealed the presence of alkaloids, phenolic compounds, and flavonoids. The phytoextract elicited significant antioxidant activity, as demonstrated by various assays, and displayed broad-spectrum antibacterial effects, producing the largest zones of inhibition (ZI) against Staphylococcus aureus, Micrococcus luteus, Escherichia coli, and Salmonella typhi, supported by MIC/MBC (minimum inhibition concentration/minimum bactericidal concentration) values indicative of bactericidal action. Notably, the phytoextract demonstrated marked cytotoxicity against various cancer cell lines, with the highest potency against MCF-7, followed by HeLa, A549, and PC-3, based on IC<inf>50</inf> (concentration required to inhibit 50%) values. Its anti-inflammatory properties, comparable to aspirin (reference standard), increased with concentration. Additionally, the phytoextract showed promise in Alzheimer's disease (AD) treatment by inhibiting AChE (acetylcholinesterase) and BuChE (butyrylcholinesterase) activities more effectively than galantamine (GA), the standard drug. The zebrafish embryo assay evidenced its excellent biocompatibility. Overall, this study offers the first comprehensive phytochemical and pharmacological evaluation of C. mariana’s methanolic extract, underscoring its potential relevance in managing inflammatory conditions. © 2025 Elsevier B.V., All rights reserved.