Small molecule inhibitor discovery for dengue virus protease using high-throughput screening.
Dengue virus (DENV), a member of mosquito-borne flavivirus genus in the Flaviviridae family, is an important human pathogen of global significance. DENV infections are the most common arbovirus infections in the world, causing more than ~300 million cases annually. Although majority of infections result in simple self-limiting disease known as dengue fever which resolve in 7-10 days, ~500,000 cases lead to more severe complications known as dengue hemorrhagic fever/dengue shock syndrome, more frequently observed in secondary infections due to an antibody-dependent enhancement mechanism, resulting in ~25,000 deaths. Currently, there are no vaccines or antiviral drug available for the treatment of DENV infections. Several viral and host proteins have been identified as potential targets for drug development. Some of the viral targets have enzyme activities that play essential roles in viral RNA replication for which in vitro high-throughput screening (HTS) assays have been developed. In this chapter, we describe an in vitro assay for the viral serine protease that has been successfully adapted to HTS format and has been used to screen several thousand compounds to identify inhibitors of the viral protease.
Department of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA.
Mark
M
Manzano
Janak
J
Padia
Radhakrishnan
R
Padmanabhan
D003716:Dengue Virus / Q000187:drug effects / Q000201:enzymology*; D055808:Drug Discovery; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D011480:Protease Inhibitors / Q000032:analysis* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D015203:Reproducibility of Results; D012697:Serine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000032:analysis* / Q000494:pharmacology; D013223:Statistics as Topic; D013997:Time Factors; D014764:Viral Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification
D011480:Protease Inhibitors; D011994:Recombinant Proteins; D054852:Small Molecule Libraries; D014764:Viral Proteins; D012697:Serine Endopeptidases