posted on 2024-02-05, 23:49authored byErnawati Arifin Giri-Rachman, Vergio V. Effendy, Muhammad H. S. Azmi, Nicholas Yamahoki, Rebecca Stephanie, Dian F. Agustiyanti, Popi H. Wisnuwardhani, Marissa Angelina, Yana Rubiyana, Reza Aditama, Ratih A. Ningrum, Andri Wardiana, Azzania Fibriani
The COVID-19 endemic
remains a global concern. The search for effective
antiviral candidates is still needed to reduce disease risk. However,
the availability of high biosafety level laboratory facilities for
drug screening is limited in number. To address this issue, a screening
system that could be utilized at lower biosafety levels remains essential.
This study aimed to develop a novel SARS-CoV-2 main protease (Mpro) dimer-based screening system (DBSS) utilizing synthetic
biology in Escherichia coli BL21(DE3).
We linked the SARS-CoV-2 Mpro with the DNA-binding domain
of AraC regulatory protein, which regulates the reporter gene expression.
Protein modeling and molecular docking showed that saquinavir could
bind to AraC-Mpro both in its monomer and dimer forms.
The constructed DBSS assay indicated the screening system could detect
saquinavir inhibitory activity at a concentration range of 4–10
μg/mL compared to the untreated control (P ≤
0.05). The Vero E6 cell assay validated the DBSS result that saquinavir
at 4–10 μg/mL exhibited antiviral activity against SARS-CoV-2.
Our DBSS could be used for preliminary screening of numerous drug
candidates that possess a dimerization inhibitor activity of SARS-CoV-2
Mpro and also minimize the use of a high biosafety level
laboratory.