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Home / Equine Research Bank / PLoS Pathog / Discovery of a novel compound with anti-venezuelan equine en...
PLoS pathogens2014; 10(6); e1004213; doi: 10.1371/journal.ppat.1004213

Discovery of a novel compound with anti-venezuelan equine encephalitis virus activity that targets the nonstructural protein 2.

Abstract: Alphaviruses present serious health threats as emerging and re-emerging viruses. Venezuelan equine encephalitis virus (VEEV), a New World alphavirus, can cause encephalitis in humans and horses, but there are no therapeutics for treatment. To date, compounds reported as anti-VEEV or anti-alphavirus inhibitors have shown moderate activity. To discover new classes of anti-VEEV inhibitors with novel viral targets, we used a high-throughput screen based on the measurement of cell protection from live VEEV TC-83-induced cytopathic effect to screen a 340,000 compound library. Of those, we identified five novel anti-VEEV compounds and chose a quinazolinone compound, CID15997213 (IC50 = 0.84 µM), for further characterization. The antiviral effect of CID15997213 was alphavirus-specific, inhibiting VEEV and Western equine encephalitis virus, but not Eastern equine encephalitis virus. In vitro assays confirmed inhibition of viral RNA, protein, and progeny synthesis. No antiviral activity was detected against a select group of RNA viruses. We found mutations conferring the resistance to the compound in the N-terminal domain of nsP2 and confirmed the target residues using a reverse genetic approach. Time of addition studies showed that the compound inhibits the middle stage of replication when viral genome replication is most active. In mice, the compound showed complete protection from lethal VEEV disease at 50 mg/kg/day. Collectively, these results reveal a potent anti-VEEV compound that uniquely targets the viral nsP2 N-terminal domain. While the function of nsP2 has yet to be characterized, our studies suggest that the protein might play a critical role in viral replication, and further, may represent an innovative opportunity to develop therapeutic interventions for alphavirus infection.
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Publication Date: 2014-06-26 PubMed ID: 24967809PubMed Central: PMC4072787DOI: 10.1371/journal.ppat.1004213Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article is about the discovery of a new compound CID15997213 with a potential to inhibit Venezuelan equine encephalitis virus (VEEV), a harmful virus that can cause encephalitis in humans and horses, by targeting the nonstructural protein 2 (nsP2).

Anti-VEEV Compound Discovery

  • The researchers started out with a high-throughput screening method to inspect a huge compound library containing 340,000 different potential components.
  • The purpose of this process was to discover new types of anti-VEEV inhibiting compounds with novel targets within the virus.
  • The selection was based on the criterion of cell protection from VEEV TC-83 virus, which causes a cytopathic effect damaging cells.
  • Out of the large library, they identified five new anti-VEEV compounds, and the quinazolinone compound CID15997213 stood out for further research due to its high inhibiting factor (IC50 = 0.84 µM).

Antiviral Effect of CID15997213

  • The antiviral effect was alphavirus-specific, appearing to inhibit VEEV and Western equine encephalitis virus, but it did not affect the Eastern equine encephalitis virus.
  • In vitro assays confirmed that the compound was able to successfully inhibit viral RNA, protein, and progeny synthesis i.e., it has the potential to prevent the virus from replicating and spreading.
  • The compound did not appear to have an antiviral activity against other types of RNA viruses.

CID15997213’s Mode of Action

  • Mutation experiments provided insights into how the compound worked: it targeted the N-terminal domain of the nsP2 protein in the virus.
  • The researchers carried out a reverse genetic approach to validate these target residues on the loci of the nsP2 protein.
  • The time of addition studies determined the compound is most effective during the middle stage of replication, when viral genome replication is at its peak.

Potential Therapeutic Impact

  • In controlled tests using mice models, the compound showed promising protective efficacy from lethal VEEV ailments at a dosage of 50 mg/kg/day.
  • This indicates that the compound could potentially be developed into a reliable therapeutic agent for VEEV infection in humans and horses.
  • The fact that it targets the viral nsP2 N-terminal domain opens up new possibilities in antiviral drug development for alphavirus infection.

Cite This Article

APA
Chung DH, Jonsson CB, Tower NA, Chu YK, Sahin E, Golden JE, Noah JW, Schroeder CE, Sotsky JB, Sosa MI, Cramer DE, McKellip SN, Rasmussen L, White EL, Schmaljohn CS, Julander JG, Smith JM, Filone CM, Connor JH, Sakurai Y, Davey RA. (2014). Discovery of a novel compound with anti-venezuelan equine encephalitis virus activity that targets the nonstructural protein 2. PLoS Pathog, 10(6), e1004213. https://doi.org/10.1371/journal.ppat.1004213

Publication

PLoS pathogens
ISSN: 1553-7374
NlmUniqueID: 101238921
Country: United States
Language: English
Volume: 10
Issue: 6
Pages: e1004213
PII: e1004213

Researcher Affiliations

Chung, Dong-Hoon
  • Departments of Microbiology and Immunology, University of Louisville, Louisville, Kentucky, United States of America; Center for Predictive Medicine for Biodefense and Emerging Infectious Diseases, University of Louisville, Louisville, Kentucky, United States of America.
Jonsson, Colleen B
  • Departments of Microbiology and Immunology, University of Louisville, Louisville, Kentucky, United States of America; Center for Predictive Medicine for Biodefense and Emerging Infectious Diseases, University of Louisville, Louisville, Kentucky, United States of America; Department of Pharmacology and Toxicology, University of Louisville, Louisville, Kentucky, United States of America.
Tower, Nichole A
  • Drug Discovery Department, Southern Research Institute, Birmingham, Alabama, United States of America.
Chu, Yong-Kyu
  • Center for Predictive Medicine for Biodefense and Emerging Infectious Diseases, University of Louisville, Louisville, Kentucky, United States of America.
Sahin, Ergin
  • Departments of Microbiology and Immunology, University of Louisville, Louisville, Kentucky, United States of America; Center for Predictive Medicine for Biodefense and Emerging Infectious Diseases, University of Louisville, Louisville, Kentucky, United States of America.
Golden, Jennifer E
  • University of Kansas Specialized Chemistry Center, Lawrence, Kansas, United States of America.
Noah, James W
  • Drug Discovery Department, Southern Research Institute, Birmingham, Alabama, United States of America.
Schroeder, Chad E
  • University of Kansas Specialized Chemistry Center, Lawrence, Kansas, United States of America.
Sotsky, Julie B
  • Center for Predictive Medicine for Biodefense and Emerging Infectious Diseases, University of Louisville, Louisville, Kentucky, United States of America.
Sosa, Melinda I
  • Drug Discovery Department, Southern Research Institute, Birmingham, Alabama, United States of America.
Cramer, Daniel E
  • Center for Predictive Medicine for Biodefense and Emerging Infectious Diseases, University of Louisville, Louisville, Kentucky, United States of America.
McKellip, Sara N
  • Drug Discovery Department, Southern Research Institute, Birmingham, Alabama, United States of America.
Rasmussen, Lynn
  • Drug Discovery Department, Southern Research Institute, Birmingham, Alabama, United States of America.
White, E Lucile
  • Drug Discovery Department, Southern Research Institute, Birmingham, Alabama, United States of America.
Schmaljohn, Connie S
  • The United States Army Medical Research Institute for Infectious Diseases, Ft. Detrick, Maryland, United States of America.
Julander, Justin G
  • Institute for Antiviral Research, Utah State University, Logan, Utah, United States of America.
Smith, Jeffrey M
  • The United States Army Medical Research Institute for Infectious Diseases, Ft. Detrick, Maryland, United States of America.
Filone, Claire Marie
  • Boston University, Boston, Massachusetts, United States of America.
Connor, John H
  • Boston University, Boston, Massachusetts, United States of America.
Sakurai, Yasuteru
  • Texas Biomedical Research Institute, San Antonio, Texas, United States of America.
Davey, Robert A
  • Texas Biomedical Research Institute, San Antonio, Texas, United States of America.

MeSH Terms

  • Animals
  • Antiviral Agents / pharmacology
  • Cell Line
  • Chlorocebus aethiops
  • Cricetinae
  • Disease Models, Animal
  • Drug Resistance, Viral / genetics
  • Encephalitis Virus, Venezuelan Equine / drug effects
  • Encephalitis Virus, Venezuelan Equine / genetics
  • Encephalomyelitis, Venezuelan Equine / drug therapy
  • Encephalomyelitis, Venezuelan Equine / virology
  • High-Throughput Screening Assays
  • Mice
  • Mice, Inbred C3H
  • Quinazolinones / pharmacology
  • Species Specificity
  • Structure-Activity Relationship
  • Vero Cells
  • Viral Plaque Assay
  • Virus Replication / drug effects

Grant Funding

  • R03 MH087448 / NIMH NIH HHS
  • U54 HG005031 / NHGRI NIH HHS
  • U54 HG005034 / NHGRI NIH HHS
  • R03MH087448-01A1 / NIMH NIH HHS

Conflict of Interest Statement

The authors have declared that no competing interests exist.

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