Screening for dual-target enzyme inhibitors derived from Gardenia jasminoides Roots, a medicinal and edible plant: Investigation of their potential anti-dementia activity

Xuanlin Liu¹Yang Zhou¹Rui Meng¹Jingyi Han¹Ming Chen¹Xiangjie Bo²Sainan Li^1*Daqian Song³Yuchi Zhang¹

• ¹Changchun Normal University, Changchun, China
• ²Northeast Normal University, Changchun, China
• ³Jilin University, Changchun, China

Gardenia jasminoides root (GJR) is a traditional Chinese plant valued for its dual functions as both a medicinal herb and an edible resource. Alzheimer's disease (AD) is a common neurodegenerative disorder in the elderly, which is irreversible and fatal. Currently, the treatment for AD mainly focuses on single-target acetylcholinesterase inhibitors, but their effect on improving the disease progression is minimal. Therefore, it is necessary to find dual-target inhibitors that can regulate inflammation (5-lipoxygenase, 5-LOX) and improve cholinergic dysfunction (acetylcholinesterase, AChE). To enhance the efficiency and accuracy of in vitro screening, receptor-ligand affinity ultrafiltration coupled with enzyme kinetics was employed for the rapid identification and characterization of active compounds. The inhibitory activities and underlying mechanisms of these compounds were further validated through biochemical assays. In addition, molecular docking and molecular dynamics simulations were conducted to evaluate target binding affinity and stability at the atomic level. The integration of experimental and computational approaches enabled a multi-dimensional elucidation of the active components and their pharmacological profiles. To overcome the limitations of conventional countercurrent chromatography—specifically its low peak capacity and separation efficiency—an offline two-dimensional chromatographic method was developed. This approach offers superior peak capacity, enhanced resolution, larger injection volume, and improved selectivity. The results demonstrate that seven active compounds, including Shanziside, Deacetylasperulosidic acid methyl ester, Gardoside, Shanzhiside methyl ester, Mussaenoside acid, Eleutheroside E, and 5-Hydroxy-3',4'-dimethoxyflavone, were successfully isolated and identified. This study offers profound insights into the active compounds of GJR and their potential anti-AD properties.This study integrates advanced screening, optimized extraction, and rigorous bioactivity assessment to provide some insights into the development of dual-target inhibitors and the advancement of plant-based food preparation methodologies.