Targeting dengue virus envelope protein: Insights from structural analysis and high-throughput screening

Area of Science:

• Virology
• Structural Biology
• Drug Discovery

Background:

• Dengue virus (DENV) is a major global health concern, causing significant morbidity and mortality.
• The DENV envelope (E) protein is crucial for viral entry into host cells by mediating membrane fusion.
• Inhibiting E protein-mediated fusion presents a viable antiviral strategy.

Purpose of the Study:

• To identify novel drug-binding sites on the DENV-2 E protein.
• To discover small molecules that inhibit DENV-2 fusion and entry.
• To establish a framework for flavivirus entry inhibitor development.

Main Methods:

• Determined three-dimensional structures of DENV-2 soluble E protein (sE) in pre- and intermediate-fusion states.
• Performed virtual screening of small molecule libraries against identified drug-binding regions.
• Utilized a high-content imaging (HCI)-based cell-cell fusion assay for high-throughput screening.
• Validated hits using infection-based assays.

Main Results:

• Identified three potential drug-binding regions on the DENV-2 sE: β-OG pocket, fusion-loop region, and stem-zipper-associated region.
• Discovered several compounds with micromolar anti-fusion activity against DENV-2.
• Narrowed down to three promising fusion inhibitors targeting the predicted sites.

Conclusions:

• The study provides a rational approach for discovering flavivirus entry inhibitors.
• Identified specific structural regions and compounds for further development of dengue antivirals.
• Highlights the potential of targeting viral fusion mechanisms for therapeutic intervention.