Identification of bioactive natural products using yeast：Application to monoacylglycerol lipase inhibitor extraction from Corydalis Rhizoma.

Abstract

Identification of bioactive principles from natural products is considered a challenging task in drug discovery. Recently, ligand fishing has been growing in interest as a sustainable strategy. In this study, a yeast-based drug discovery strategy was investigated to screen and fish active ingredients from natural products. Human monoacylglycerol lipase (MAGL) was first displayed on the cell wall of Pichia pastoris. The expression of MAGL on the cell surface was confirmed by immunofluorescence analysis. The activity toward 7-HCA which was consistent with free enzymes in solution. Recombinant yeast strains were used to screen the potential inhibitors from traditional Chinese medicines. Preliminary screening showed that the extracts of 12 herbs showed inhibition on MAGL activity, among which Corydalis Rhizoma had the highest inhibition rate of 60.66 ± 2.44%. Recombinant yeast fishing combined with HPLC-Q-TOF-MS/MS analysis was further used to identify the potential MAGL inhibitors. Finally, dehydrocorydaline (DHC) in Corydalis Rhizoma was determined as a ligand to MAGL with the IC50 value at 154.7 μΜ. Corydalis Rhizoma has been used as a pain reliever in TCM. Intraperitoneal injection of 7 mg kg- 1 DHC in chronic constriction injury model rats significantly attenuated the mechanical allodynia and thermal hyperalgesia. Meanwhile, 2-arachidonoylglycerol, the major MAGL substrate in the brain, was significantly increased both in the hippocampus and striatum. In conclusion, yeast-based ligand fishing combined with HPLC-Q-TOF-MS/MS is a powerful strategy for drug discovery in complex mixtures and DHC from Corydalis Rhizoma was confirmed with high inhibitory activity to MAGL either in vitro or in vivo .

• Authors: Jie Mei, Ruilin Guo, Fan Zhang, Haiyan Zhang, Xinping Yang, Boyang Yu, Jihua Liu, Xiufeng Liu
• Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie (Biomed Pharmacother) Vol. 149 Pg. 112798 (May 2022) ISSN: 1950-6007 [Electronic] France
• PMID: 35286964 (Publication Type: Journal Article)
• Copyright: Copyright © 2022 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Chemical References

• Biological Products
• Enzyme Inhibitors
• Ligands
• Monoglycerides
• Monoacylglycerol Lipases

Topics

• Animals
• Biological Products (pharmacology)
• Corydalis (metabolism)
• Enzyme Inhibitors (pharmacology)
• Ligands
• Monoacylglycerol Lipases
• Monoglycerides
• Rats
• Saccharomyces cerevisiae (metabolism)
• Tandem Mass Spectrometry