Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comment
. 2021 Jul;12(7):586-591.
doi: 10.1007/s13238-020-00803-w. Epub 2020 Nov 18.

Repurposing FDA-approved drugs for SARS-CoV-2 through an ELISA-based screening for the inhibition of RBD/ACE2 interaction

Wenyu Fu #  1 , Yujianan Chen #  1 , Kaidi Wang #  1 , Aubryanna Hettinghouse  1 , Wenhuo Hu  2 , Jing-Quan Wang  3 , Zi-Ning Lei  3 , Zhe-Sheng Chen  3 , Kenneth A Stapleford  4 , Chuan-Ju Liu  5   6
Affiliations
Comment

Repurposing FDA-approved drugs for SARS-CoV-2 through an ELISA-based screening for the inhibition of RBD/ACE2 interaction

Wenyu Fu et al. Protein Cell. 2021 Jul.
No abstract available

PubMed Disclaimer

Figures

Figure 1
Figure 1
VP, NAC and TPR dose-dependently inhibit the binding of the RBD of SARS-CoV-2 Spike protein to ACE2 and cell surface. (A) Biotinylated RBD (2 ng/mL) and serial 10-fold dilutions of VP were added into 96-well plates coated with 1 µg/mL ACE2, ACE2 bound RBD was detected by avidin-HRP and TMB substrate. (B) The serial 10-fold dilutions of VP were added into 96-well plates coated with 1 µg/mL ACE2 and incubated for 1 h, followed by addition of biotinylated RBD (2 ng/mL) for 1 h, and ACE2 bound RBD was detected by avidin-HRP and TMB substrate. (C and E) Biotinylated RBD (2 ng/mL) and NAC (C) or TPR (E) mixture was added into 96-well plates coated with 1 µg/mL ACE2, and ACE2 bound RBD was detected by avidin-HRP and TMB substrate. (D and F) Biotinylated RBD (2 ng/mL) was incubated with serial 10-fold dilutions of NAC (D) or TPR (F), respectively, for 1 h at RT before addition into 96-well plates coated with 1 µg/mL ACE2, and RBD bound to ACE2 was detected by avidin-HRP and TMB substrate. (A–F), data are shown as mean ± SD, n = 3 biological replicates. (G) A serial dilutions of VP were incubated with cells for 1 h before addition of RBD for 30 min, followed by flow cytometry analysis. (I and K) RBD was incubated with a serial dilutions of NAC or TPR for 1 h and the RBD-drug mixture was added into the cells and incubated on ice for 30 min. (H, J and L) RBD and serial dilutions of drugs were added into the cells at the same time and incubated on ice for 30 min before flow cytometry analysis. (G–L), experiments were repeated three times and yielded similar results, and representative images shown
Figure 2
Figure 2
The anti-viral activities of VP, NAC and TPR against pseudotyped and authentic SARS-CoV-2 in vitro. (A) hACE2 overexpressing HEK 293T cells (4.0 × 104 cells/well) were pretreated with VP for 2 h, then infected with pseudoviruses; or pseudoviruses were incubated with NAC or TPR for 2 h before adding into the hACE2 overexpressing HEK 293T cells. After 4 h incubation, medium was replaced with fresh DMEM with 10% FBS and cells were incubated for an additional 24 h. Neutralization potencies of VP, NAC or TPR were evaluated by luciferase assay system. (B) VP, NAC or TPR and pseudovirus mixture were added to hACE2 overexpressing HEK 293T cells concurrently. The cells were then processed as described in (A). (C and D) The mixture of authentic SARS-CoV-2 virus at an MOI of 0.1 and VP (C), or NAC or TPR (D) was added to Vero E6 or Calu-3 cells. After 24 h incubation, the virus yield in the infected cell supernatants was qualified by RT-qPCR. (E) Representative images of virus infection with VP, NAC, and TRP at 24 h post infection. Cells were fixed with 4% paraformaldehyde for 1 h, stained with DAPI, and imaged. Data represent the mean and standard error of the mean. All experiments were performed in triplicate. **P < 0.01; t-test
See this image and copyright information in PMC

Comment on

References

    1. Castañeda-Arriaga R, Pérez-González A, Galano A. Chemical Protectors against the Toxic Effects of Paracetamol (Acetaminophen) and Its Meta Analogue: Preventing Protein Arylation. ACS Omega. 2018;3:18582–18591. doi: 10.1021/acsomega.8b02943. - DOI
    1. De Flora S, Grassi C, Carati L. Attenuation of influenza-like symptomatology and improvement of cell-mediated immunity with long-term N-acetylcysteine treatment. Eur Respir J. 1997;10:1535–1541. doi: 10.1183/09031936.97.10071535. - DOI - PubMed
    1. Geiler J, Michaelis M, Naczk P, Leutz A, Langer K, Doerr HW, Cinatl J., Jr N-acetyl-L-cysteine (NAC) inhibits virus replication and expression of pro-inflammatory molecules in A549 cells infected with highly pathogenic H5N1 influenza A virus. Biochem Pharmacol. 2010;79:413–420. doi: 10.1016/j.bcp.2009.08.025. - DOI - PubMed
    1. Ghezzi P, Ungheri D. Synergistic combination of N-acetylcysteine and ribavirin to protect from lethal influenza viral infection in a mouse model. Int J Immunopathol Pharmacol. 2004;17:99–102. doi: 10.1177/039463200401700114. - DOI - PubMed
    1. Gordon DE, Jang GM, Bouhaddou M, Xu J, Obernier K, White KM, O’Meara MJ, Rezelj VV, Guo JZ, Swaney DL, et al. A SARS-CoV-2 protein interaction map reveals targets for drug repurposing. Nature. 2020;583:459–468. doi: 10.1038/s41586-020-2286-9. - DOI - PMC - PubMed

Substances