Discovery and evaluation of active compounds from Xuanfei Baidu Formula against COVID-19 via SARS-Cov-2 Mpro

Abstract

Abstract Background The coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus (SARS-CoV-2) is still a widespread concern. As one of the effective traditional Chinese medicine (TCM) formula, Xuanfei Baidu formula (XFBD) shows significant efficacy for treatment of COVID-19 patients. However, its antiviral compounds and mechanism are still unclear. Purpose: In this study, we explored the bioactive compounds of XFBD and its antiviral mechanism by integrating computational analysis and experimental testing. Methods Aiming at the SARS-CoV-2 main protease (Mpro), as a key target in virus replication, the fluorescence resonance energy transfer (FRET) assay was built to screen out satisfactory natural inhibitors from XFBD. The surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) were undertaken to verify the binding affinity of Mpro-ligand. Omicron BA.1.1 and BA.2 variants were used to evaluate the antiviral activity of the focused compounds in non-cytotoxicity concentrations. For introducing the molecular mechanism, computational modeling and NMR spectra were employed to predict the binding mode and binding site of Mpro-ligand. Results From a library of 83 natural compounds, acteoside, licochalcone B, licochalcone D, linoleic acid, and physcion showed the satisfactory inhibition effect on Mpro with IC50 from 1.93 to 42.96 µM, which were further verified by SPR. Showing the excellent binding affinity, acteoside was witnessed to gain valuable insights into the thermodynamic signatures by ITC and presented antiviral activity on Omicron BA.1.1 and BA.2.3 variants in vitro. The results revealed that acteoside inhibited Mpro via forming the hydrogen bond between 7-H of acteoside and Mpro. Conclusion Acteoside is regarded as a representative active natural compound in XFBD to inhibit replication of SARS-CoV-2, which provides the antiviral evidence and some insight into the identifications of SARS-CoV-2 Mpro natural inhibitors.