Abiraterone Acetate Attenuates SARS-CoV-2 Replication by Interfering with the Structural Nucleocapsid Protein

Abstract

Since infection with severe acute respiratory syndrome CoV-2 (SARS-CoV-2) was first reported in Wuhan City, China in December 2019 (Wu et al., 2020;Zhou et al., 2020a), the pandemic coronavirus disease 2019 (COVID-19) has caused serious challenges for the public health and medical systems worldwide (https://covid19.who.int/). SARS-CoV-2 is an enveloped virus with a plus-stranded RNA genome (varies from 29.8 kb to 29.9 kb) and belongs to lineage B beta-coronaviruses (Khailany et al., 2020). SARS-CoV-2 maintains its structure with four major structural protein complexes: spike (S) glycoprotein, nucleocapsid (N) phosphoprotein, membrane (M) glycoprotein and small envelope (E) glycoprotein (Satarker and Nampoothiri, 2020). It is known that angiotensin-converting enzyme 2 (ACE2) is a receptor for SARS-CoV-2 S protein for mediating viral attachment/membrane fusion and that transmembrane protease serine 2 (TMPRSS2) contributes to viral entry into host cells (Datta et al., 2020;Matsuyama et al., 2020;Zeidler and Karpinski 2020). In this regard, the therapeutic strategy against SARS-CoV-2 infection targeting the entry process is directed towards the development of neu- 427The drug repurposing strategy has been applied to the development of emergency COVID-19 therapeutic medicines. Current drug repurposing approaches have been directed against RNA polymerases and viral proteases. Recently, we found that the inhibition of the interaction between the SARS-CoV-2 structural nucleocapsid (N) and spike (S) proteins decreased viral replication. In this study, drug repurposing candidates were screened by in silico molecular docking simulation with the SARS-CoV-2 structural N protein. In the ChEMBL database, 1994 FDA-approved drugs were selected for the in silico virtual screening against the N terminal domain (NTD) of the SARS-CoV-2 N protein. The tyrosine 109 residue in the NTD of the N protein was used as the center of the ligand binding grid for the docking simulation. In plaque forming assays performed with SARS-CoV-2 infected Vero E6 cells, atovaquone, abiraterone acetate, and digoxin exhibited a tendency to reduce the size of the viral plagues without affecting the plaque numbers. Abiraterone acetate significantly decreased the accumulation of viral particles in the cell culture supernatants in a concentration-dependent manner. In addition, abiraterone acetate significantly decreased the production of N protein and S protein in the SARS-CoV-2-infected Vero E6 cells. In conclusion, abiraterone acetate has therapeutic potential to inhibit the viral replication of SARS-CoV-2.

Differential effect of SARS-CoV-2 infection on stress granule formation in Vero and Calu-3 cells

“…To estimate viral titers, we performed plaque assays on Vero E6 cells, as described previously ( Kim et al, 2021b , 2022a ; Park et al, 2021a ). In brief, Vero E6 cells (7 × 10 5 cells/well) were cultured in 6-well plates (Corning Inc., Corning, NY, United States) for 12 h. Cells were then washed with phosphate-buffered saline (PBS) and infected with 10-fold serial dilutions of virus culture supernatants.…”

“…Monoclonal antibody against the SARS-CoV-2 N protein (anti-SARS-CoV-2 N mAb) was prepared from ascitic fluid collected from BALB/c mice injected with hybridoma cells (clone 1G10C4 mAb), as previously described ( Kim et al, 2021a , 2022a , 2022b ). Rabbit anti-SARS-CoV-2 N protein polyclonal antibody (anti-SARS-CoV-2 N Ab, Catalog no.…”

The Outcomes of Sodium-Glucose Co-transporter 2 Inhibitors (SGLT2I) on Diabetes-Associated Neuropathy: A Systematic Review and meta-Analysis

“…Computational tools and meta-analysis are gold standards in diseases epidemiology, treatment and control ( Kandeel et al, 2021 ; Kandeel and El-Deeb, 2022a ; Kandeel and El-Deeb, 2022b ; Burayk et al, 2022 ; Kandeel et al, 2022 ; Kim et al, 2022 ). DN is the most encountered microvascular complication of diabetes.…”

Production of a Monoclonal Antibody to the Nucleocapsid Protein of SARS-CoV-2 and Its Application to ELISA-Based Detection Methods with Broad Specificity by Combined Use of Detector Antibodies

“…The cells were washed and infected with each SARS-CoV-2 variant at a multiplicity of infection (MOI) of 0.01 and incubated for 1 h at 37 °C with shaking every 20 min. After incubation, the medium was replaced with 2 mL of DMEM containing 2% FBS and cultured for 72 h. Thereafter, the cell culture supernatants were collected, and the viruses were quantified by plaque assay on Vero E6 cells as described previously [ 26 , 27 , 28 ]. After determining the pfu number, the virus stocks were stored at −70 °C for future use.…”