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Development of a NS2B/NS3 protease inhibition assay using AlphaScreen® beads for screening of anti-dengue activities

Abduraman MA 1 , Hariono M 2 , Yusof R 3 , Rahman NA 4 , Wahab HA 1 , Tan ML 1

Affiliations 

  • 1 Malaysian Institute of Pharmaceuticals & Nutraceuticals, National Institutes of Biotechnology Malaysia (NIBM), Ministry of Science, Technology and Innovation Malaysia, Pulau Pinang, Malaysia
  • 2 Pharmaceuticals Drug Simulation Laboratory (PhDS), School of Pharmaceutical Sciences, Universiti Sains Malaysia, Pulau Pinang, Malaysia
  • 3 Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia
  • 4 Department of Chemistry, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia
Heliyon, 2018 Dec;4(12):e01023.
PMID: 30560214 DOI: 10.1016/j.heliyon.2018.e01023

Abstract

Background: Dengue infection is an endemic infectious disease and it can lead to dengue fever, dengue hemorrhagic fever, and/or dengue shock syndromes. Dengue NS2B/NS3 protease complex is essential for viral replication and is a primary target for anti-dengue drug development. In this study, a NS2B/NS3 protease inhibition assay was developed using AlphaScreen® beads and was used to screen compounds for their protease inhibition activities.

Methods: The assay system utilized a known NS2B/NS3 peptide substrate, a recombinant of NS2B/NS3 protease with proprietary StrepTactin® donor and nickel chelate acceptor beads in 384-well format.

Results: The optimized assay to screen for NS2B/NS3 protease inhibitors was demonstrated to be potentially useful with reasonable z' factor, coefficient variance and signal to background ratio. However, screening of synthesized thioguanine derivatives using the optimized AlphaScreen® assay revealed weak NS2B/NS3 inhibition activities.

Conclusion: The AlphaScreen® assay to screen for NS2B/NS3 protease inhibitors is potentially applicable for high throughput screening.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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