In Silico Design of miniACE2 Decoys with In Vitro Enhanced Neutralization Activity against SARS-CoV-2, Encompassing...

Area of Science:

• Virology and Immunology
• Biopharmaceutical Development
• Computational Biology

Background:

• The COVID-19 pandemic continues to pose global health and economic challenges, exacerbated by the continuous evolution of SARS-CoV-2 variants.
• Immunocompromised individuals are susceptible to prolonged infections, contributing to viral evolution and the emergence of new variants.
• Existing vaccines and therapies face challenges in effectively neutralizing novel SARS-CoV-2 strains.

Purpose of the Study:

• To explore miniACE2 decoys as a novel therapeutic strategy against SARS-CoV-2 variants.
• To design and develop blocking proteins (BPs) with enhanced binding affinity to the SARS-CoV-2 receptor-binding domain (RBD) of multiple variants.
• To evaluate the in vitro neutralizing efficacy of these miniACE2 decoys.

Main Methods:

• In silico design and molecular dynamics simulations were employed to engineer miniACE2 decoys.
• Blocking proteins (BPs) were designed for stronger binding affinity to the RBD of various SARS-CoV-2 variants compared to natural ACE2.
• The BPs were expressed in *E. coli* and their neutralizing effects were assessed through in vitro assays.

Main Results:

• Engineered miniACE2 blocking proteins demonstrated superior binding affinity to the RBD of multiple SARS-CoV-2 variants.
• In vitro testing showed promising neutralizing effects of the developed BPs against various strains.
• MiniACE2 BP9 achieved a potent average IC50 of 4.9 µg/mL across the Wuhan strain, Mu, Omicron BA.1, and BA.2 variants.

Conclusions:

• MiniACE2 BP9 exhibits significant neutralizing capability against a range of SARS-CoV-2 variants at low concentrations.
• BP9 represents a promising therapeutic candidate for combating current and future SARS-CoV-2 variants.
• This novel decoy strategy holds potential as an emergency biopharmaceutical for COVID-19 treatment.