Microplate-based assay for identifying small molecules that bind a specific intersubunit interface within the...

Area of Science:

• Virology
• Drug Discovery
• Biochemistry

Background:

• Current HIV-1 therapies are not curative and face challenges from drug-resistant strains.
• The HIV-1 capsid protein is a conserved viral target with critical roles in replication.
• Existing capsid inhibitors lack sufficient potency for therapeutic development.

Purpose of the Study:

• To develop a robust assay for identifying potent small-molecule inhibitors of the HIV-1 capsid.
• To facilitate the discovery of novel therapeutic agents against HIV-1.

Main Methods:

• Developed a scintillation proximity assay (SPA) to screen for compounds targeting a specific HIV-1 capsid intersubunit interface.
• The SPA utilizes competitive displacement of a known capsid-binding inhibitor.
• Validated the assay with a signal-to-noise ratio >9 and a Z factor >0.8.

Main Results:

• A pilot screen of 2,400 druglike compounds yielded a hit rate of 1.8%.
• The developed SPA demonstrates suitability for large-scale screening of compound libraries.
• Identified potential novel HIV-1 capsid ligands with antiviral activity.

Conclusions:

• The novel SPA is effective for identifying potent HIV-1 capsid inhibitors.
• This assay can accelerate the development of new anti-HIV-1 drugs.
• The findings support targeting the HIV-1 capsid for improved HIV management and resistance control.