Efficient and fast screening and separation based on computer aided screening and complex chromatography method for 5-lipoxygenase inhibitors from Poria cocos

Poria cocos (Schw.) Wolf ( P. cocos ) is a traditional Chinese medicinal materia that medicine food homologyl. In this study, based on the β-Amyloid deposition hypothesis of AD, a fast and efficient methods that ultrafiltration-liquid chromatography-mass spectrometry (UF-LC-MS) and molecular docking were developed for the rapid screening and identification of anti-5-lipoxygenase (anti-5-LOX) enzyme active ingredient of P. cocos ethyl acetate extract. Continuous counter-current chromatography (CCC) combined with semi-preparative high performance liquid chromatography (Semi-preparative HPLC) was developed for targeted separation of active components. Five potential 5-LOX inhibitors were screened by ultrafiltration affinity assay in P. cocos . The molecular docking simulation results are consistent with the ultrafiltration experimental results, which further verifies the accuracy of the experiment. Subsequently, five high-purity active ingredients (Tumulosic acid, Polyporenic acid C, 3- Epi -dehydrotumulosic acid, Pachymic acid and Dehydrotrametenolic acid) could be isolated by the established separation method, with the purities of 95.86%, 96.35%, 97.22%, 98.43% and 99.00%, respectively. The established continuous countercurrent chromatography can effectively improve the yield and purity of low content active ingredients in traditional Chinese medicine, and provide ideas and methods for the targeted separation of active ingredients. The methods of screening and isolating active ingredients established in this paper will be helpful for the development of new therapeutic drugs for Alzheimer's disease (AD).