Kanda_2026_Chem.Biodivers_23_e02600

Alzheimer's disease (AD) progressively impairs memory and cognition. Luffa acutangula (LA), rich in triterpenoids, fatty acids, and iridoid glycosides, exerts anti-Alzheimer's effects by inhibiting acetylcholinesterase (AChE). This study aims to isolate bioactive compounds from LA fruits, formulate and optimize a nanoemulsion to improve brain targeting, and assess its anti-Alzheimer efficacy through in vitro, in vivo, and in silico approaches. LA was extracted using a 1:1 hydro-ethanol mixture and subsequently underwent chromatographic isolation to yield four major constituents: oleanolic acid, stearic acid, cucurbitacin H, and acutoside C. A multiple nanoemulsion (containing a mixture of isolated constituents) was formulated using Box-Behnken design and characterized for particle size, viscosity, entrapment efficiency, and FTIR compatibility. In vivo studies were conducted in scopolamine-induced memory-impaired mice using Hebb-William's Maze (HWM) and Cook's pole climbing tests. AChE activity was biochemically assessed, followed by histopathological brain analysis. Molecular docking was used to predict ligand-AChE binding affinities. The optimized L. acutangula loaded multiple nanoemulsion (LAMN) exhibited an average particle size of 142.1 nm, viscosity of 60 cP, and entrapment efficiency (EE) of 96.8 %. It decreased learning scores in the HWM and shortened latency time in Cook's pole-climbing test in memory-impaired mice. AChE levels were significantly reduced in LAMN-treated groups, correlating with histological evidence of hippocampal protection. Molecular docking revealed strong AChE binding, particularly by oleanolic acid (-10.5 kcal/mol), comparable to donepezil (-10.7 kcal/mol). LAMN offers a promising phytopharmaceutical intervention for AD, with multitargeted neuroprotective effects mediated through AChE inhibition and improved drug delivery across the blood-brain barrier. The findings support further clinical development of LA-based nanoformulations for AD management.