(In this application, we propose a multi-disciplinary approach for identification and further development of lead molecules already in hand against DENV serine protease (DENV NSSpro) and RNA-dependent RNA polymerase (DENV RdRP). The product development plan would involve completion of following three specific aims and milestones.
In aim 1, we propose to identify new inhibitors of the viral NS3 protease and compounds that block NS3/NS5 interaction by high throughput screening (HTS) and by virtual screening. The novel series of compounds have been have been identified to date as promising protease inhibitors. Compounds will be further optimized by iterative medicinal and combinatorial chemistry, molecular modeling, to obtain potent and selective drug-like lead molecules.
In aim 2, a novel and potent antiviral agent: that we termed 7DA6MEPN that targets the NS5 RdRP has been identified to inhibit DENV replication. Additional analogues of 7-deaza-6-methylpurine ribonucleoside will be synthesized and evaluated to further improve the efficacy and therapeutic index of this promising antiviral agent.
In aim 3, compounds identified and synthesized under aims 1 and 2 will be further evaluated for inhibitory potency by in vitro enzyme assays (IC50 values), cytotoxicity assays (CC50 values), reporter replicon-based as well as virus infectivity assays (EC50) in mammalian cell culture. Selected compounds showing inhibition in the nM range will be analyzed by ADME/TOX assays.

Public Health Relevance

Dengue viruses (DENV), members of mosquito-borne flaviruses of Flaviviridae family, are included in the NIAID Category A Priority Pathogen as they cause frequent epidemics and serious illness with considerable morbidity and mortality. Two-thirds of human populations live in areas at risk from dengue virus infections. There is no vaccine or antiviral therapeutics currently available for human use.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01AI082068-02
Application #
7932902
Study Section
Special Emphasis Panel (ZAI1-MMT-M (J2))
Program Officer
Tseng, Christopher K
Project Start
2009-09-17
Project End
2012-08-31
Budget Start
2010-09-01
Budget End
2012-08-31
Support Year
2
Fiscal Year
2010
Total Cost
$543,312
Indirect Cost
Name
Georgetown University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
049515844
City
Washington
State
DC
Country
United States
Zip Code
20057
Balasubramanian, Anuradha; Manzano, Mark; Teramoto, Tadahisa et al. (2016) High-throughput screening for the identification of small-molecule inhibitors of the flaviviral protease. Antiviral Res 134:6-16
Boonyasuppayakorn, Siwaporn; Reichert, Erin D; Manzano, Mark et al. (2014) Amodiaquine, an antimalarial drug, inhibits dengue virus type 2 replication and infectivity. Antiviral Res 106:125-34
Boonyasuppayakorn, Siwaporn; Padmanabhan, Radhakrishnan (2014) Construction of plasmid, bacterial expression, purification, and assay of dengue virus type 2 NS5 methyltransferase. Methods Mol Biol 1138:361-73
Lai, Huiguo; Teramoto, Tadahisa; Padmanabhan, Radhakrishnan (2014) Construction of dengue virus protease expression plasmid and in vitro protease assay for screening antiviral inhibitors. Methods Mol Biol 1138:345-60
Manzano, Mark; Padia, Janak; Padmanabhan, Radhakrishnan (2014) Small molecule inhibitor discovery for dengue virus protease using high-throughput screening. Methods Mol Biol 1138:331-44
Lai, Huiguo; Dou, Dengfeng; Aravapalli, Sridhar et al. (2013) Design, synthesis and characterization of novel 1,2-benzisothiazol-3(2H)-one and 1,3,4-oxadiazole hybrid derivatives: potent inhibitors of Dengue and West Nile virus NS2B/NS3 proteases. Bioorg Med Chem 21:102-13
Sztuba-Solinska, Joanna; Teramoto, Tadahisa; Rausch, Jason W et al. (2013) Structural complexity of Dengue virus untranslated regions: cis-acting RNA motifs and pseudoknot interactions modulating functionality of the viral genome. Nucleic Acids Res 41:5075-89
Lai, Huiguo; Sridhar Prasad, G; Padmanabhan, Radhakrishnan (2013) Characterization of 8-hydroxyquinoline derivatives containing aminobenzothiazole as inhibitors of dengue virus type 2 protease in vitro. Antiviral Res 97:74-80
Tiew, Kok-Chuan; Dou, Dengfeng; Teramoto, Tadahisa et al. (2012) Inhibition of Dengue virus and West Nile virus proteases by click chemistry-derived benz[d]isothiazol-3(2H)-one derivatives. Bioorg Med Chem 20:1213-21
Ezgimen, Manolya; Lai, Huiguo; Mueller, Niklaus H et al. (2012) Characterization of the 8-hydroxyquinoline scaffold for inhibitors of West Nile virus serine protease. Antiviral Res 94:18-24

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