High-throughput assay using a GFP-expressing replicon for SARS-CoV drug discovery

Area of Science:

• Virology
• Drug Discovery
• Molecular Biology

Background:

• Severe acute respiratory syndrome (SARS) is caused by SARS-CoV.
• Rapid screening assays are crucial for identifying antiviral drug candidates.

Purpose of the Study:

• To develop a high-throughput screening assay for SARS-CoV replication inhibitors.
• To identify small molecule compounds with anti-SARS-CoV activity.

Main Methods:

• Utilized a cell line expressing a SARS-CoV subgenomic replicon.
• Screened 7035 small molecule compounds using the developed assay.
• Validated antiviral activity through plaque reduction assays and target identification.

Main Results:

• Identified 7 compounds exhibiting anti-SARS-CoV activity.
• Demonstrated inhibition of SARS-CoV replication, viral protein accumulation, and RNA copy number.
• Validated the target of one compound (C12344) by generating resistant cells.

Conclusions:

• The developed assay is effective for high-throughput screening of SARS-CoV inhibitors.
• The identified compounds show promise as potential therapeutic agents for SARS.
• These compounds can serve as valuable tools for studying SARS-CoV replication mechanisms.