Receptor-ligand affinity-based screening and isolation of water-soluble 5-lipoxygenase inhibitors from Phellinus igniarius

•1. Water extraction and alcohol precipitation yielded Phellinus igniarius extracts.•2. Dual probing of compound activity by UF-LC combined with molecular docking.•3. Aqueous extraction was optimized using the response surface methodology.•4. Target compounds were isolated using HSCCC combined with semi-preparative LC.•5. Purity of all three target compounds exceeded 98%. We developed an efficient combination method for extraction, biological activity screening, and preparation of 5-lipoxygenase inhibitors from Phellinus igniarius. 5-Lipoxygenase inhibitors were rapidly screened using ultrafiltration-liquid chromatography based on the receptor-ligand affinity. Parameters such as extraction time, extraction times, and temperature as well as liquid–solid ratio were optimized using response surface methodology to maximize the total yield of the three target compounds. Next, bioactive ingredients were isolated using high-speed countercurrent chromatography and semi-preparative liquid chromatography. Three active ingredients, phellibaumin E, protocatechuic aldehyde, and osmundacetone, were obtained via ultrafiltration-liquid chromatography. Subsequently, the potential anti-dementia effects of the obtained bioactive compounds were verified using molecular docking assays. The above-mentioned target compounds, with purities of 98.82%, 98.89%, and 99.51%, respectively, were separated using a two-phase solvent system consisting of n-hexane–ethyl acetate-ethanol–water (2.5:2:0.75:3, v/v/v/v) coupled with semi-preparative liquid chromatography.