Microarray compound screening (microARCS) to identify inhibitors of HIV integrase

Area of Science:

• Biochemistry
• Virology
• Drug Discovery

Background:

• Human immunodeficiency virus (HIV) integrase is a critical target for antiviral therapy.
• Developing high-throughput screening methods is essential for identifying novel HIV integrase inhibitors.

Purpose of the Study:

• To develop and validate a novel high-throughput strand transfer assay for identifying HIV integrase inhibitors.
• To screen a large library of compounds using the developed assay.

Main Methods:

• Microarray Compound Screening (microARCS) technology utilizing agarose matrices.
• Enzyme-linked immunosorbent assay (ELISA) on a SAM membrane with biotinylated donor DNA and FITC-labeled target DNA.
• Detection of integrase activity using alkaline phosphatase (AP) and AttoPhos substrate in an agarose gel.
• Imaging of the AttoPhos gel to identify potential inhibitors as dark spots.

Main Results:

• Successful development of a novel high-throughput HIV integrase strand transfer assay.
• Screening of approximately 250,000 compounds identified novel integrase inhibitors.
• Inhibitors from diverse structural classes were discovered.

Conclusions:

• The microARCS-based assay is effective for high-throughput screening of HIV integrase inhibitors.
• This assay facilitates the discovery of novel antiviral compounds.
• The identified compounds represent promising candidates for further development as HIV therapeutics.