Screening and evaluation of anti-SARS-CoV-2 components from Ephedra sinica by ACE2/CMC-HPLC-IT-TOF-MS approach
- 19 February 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Analytical and Bioanalytical Chemistry
- Vol. 413 (11), 2995-3004
- https://doi.org/10.1007/s00216-021-03233-7
Abstract
Traditional Chinese medicines played an important role in the treatment of COVID-19 in 2020. Ephedra sinica, one of the major constituent herbs of multi-component herbal formula, has been widely used to treat COVID-19 in China. However, its active components are still unclear. The objectives of this study are to screen and evaluate active components from the traditional Chinese medicine Ephedra sinica for the treatment of COVID-19. In our study, we established an ACE2/CMC bioaffinity chromatography model, and then developed an ACE2/CMC-HPLC-IT-TOF-MS system for the active compounds screening and identification from Ephedra sinica extract. We performed molecular docking and surface plasmon resonance (SPR) assays to assess the binding characteristics (binding mode and KD value). We used CCK-8 staining to assess the toxicity of screened compounds, and also used SARS-CoV-2 pseudovirus to observe the viropexis effect of screened compounds in ACE2h cells. In this current work, one fraction was fished out, separated and identified as ephedrine (EP), pseudoephedrine (PEP), and methylephedrine (MEP). Binding assays showed that the three compounds could bind with ACE2 in a special way to some amino acid residues, similar to the way SARS-CoV-2 bound with ACE2. Additionally, the three compounds, especially EP, can inhibit the entrance of SARS-CoV-2 spike pseudovirus into ACE2h cells because they can reduce the entrance ratio of pseudovirus in the pseudovirus model. Overall, the ACE2/CMC-HPLC-IT-TOF-MS system was established and verified to be suitable for ACE2-targeted bioactive compound screening. EP, PEP, and MEP with ACE2-binding features were screened out from Ephedra sinica, and acted as blockers inhibiting SARS-CoV-2 spike pseudovirus entering ACE2h cells. Graphical abstractKeywords
Funding Information
- National Natural Science Foundation of China (81973278 and 81930096)
- China Postdoctoral Science Foundation (2019T120923 and 2018M641003)
- National Science Foundation of Shaanxi Province (2020SF-309)
This publication has 36 references indexed in Scilit:
- A direct ionization mass spectrometry-based approach for differentiation of medicinal Ephedra speciesJournal of Pharmaceutical and Biomedical Analysis, 2016
- Metabolomics based on liquid chromatography with mass spectrometry reveals the chemical difference in the stems and roots derived from Ephedra sinicaJournal of Separation Science, 2015
- Pharmacokinetic comparisons of five ephedrine alkaloids following oral administration of four different Mahuang–Guizhi herb-pair aqueous extracts ratios in ratsJournal of Ethnopharmacology, 2014
- Pharmacological tools for the development of traditional Chinese medicineTrends in Pharmacological Sciences, 2013
- [New method for analyzing pharmacodynamic material basis of traditional Chinese medicines by using specific knockout technology with monoclonal antibodies].2013
- Anti‐HBV effect of individual traditional Chinese herbal medicine in vitro and in vivo: an analytic reviewJournal of Viral Hepatitis, 2013
- A vascular smooth muscle/cell membrane chromatography–offline-gas chromatography/mass spectrometry method for recognition, separation and identification of active components from traditional Chinese medicinesJournal of Chromatography A, 2009
- Plant-derived natural product research aimed at new drug discoveryJournal of Natural Medicines, 2008
- Anti‐influenza agents from plants and traditional Chinese medicinePhytotherapy Research, 2006
- Enzymatic activity and chromatographic characteristics of the cell membrane immobilized on silica surfaceChinese Science Bulletin, 1999