FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
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.
Get Full Text
Publication Date: 2014-06-26 PubMed ID: 24967809PubMed Central: PMC4072787DOI: 10.1371/journal.ppat.1004213Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • N.I.H.
  • Extramural
  • Research Support
  • Non-U.S. Gov't

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.

References

This article includes 37 references
  1. Weaver SC, Barrett AD. Transmission cycles, host range, evolution and emergence of arboviral disease.. Nat Rev Microbiol 2004 Oct;2(10):789-801.
    pmc: PMC7097645pubmed: 15378043doi: 10.1038/nrmicro1006google scholar: lookup
  2. Aguilar PV, Estrada-Franco JG, Navarro-Lopez R, Ferro C, Haddow AD, Weaver SC. Endemic Venezuelan equine encephalitis in the Americas: hidden under the dengue umbrella.. Future Virol 2011;6(6):721-740.
    pmc: PMC3134406pubmed: 21765860doi: 10.2217/fvl.11.5google scholar: lookup
  3. Paessler S, Weaver SC. Vaccines for Venezuelan equine encephalitis.. Vaccine 2009 Nov 5;27 Suppl 4:D80-5.
    pmc: PMC2764542pubmed: 19837294doi: 10.1016/j.vaccine.2009.07.095google scholar: lookup
  4. Fine DL, Jenkins E, Martin SS, Glass P, Parker MD, Grimm B. A multisystem approach for development and evaluation of inactivated vaccines for Venezuelan equine encephalitis virus (VEEV).. J Virol Methods 2010 Feb;163(2):424-32.
    pmc: PMC2815040pubmed: 19903494doi: 10.1016/j.jviromet.2009.11.006google scholar: lookup
  5. Berge TO, Banks IS, Tigertt WD. Attenuation of venezuelan equine encephalomyelitis virus by in vitro cultivation in guinea-pig heart cells. Am J Epidemiol 73: 209–218.
  6. Pittman PR, Makuch RS, Mangiafico JA, Cannon TL, Gibbs PH, Peters CJ. Long-term duration of detectable neutralizing antibodies after administration of live-attenuated VEE vaccine and following booster vaccination with inactivated VEE vaccine.. Vaccine 1996 Mar;14(4):337-43.
    pubmed: 8744562doi: 10.1016/0264-410x(95)00168-zgoogle scholar: lookup
  7. Jahrling PB, Stephenson EH. Protective efficacies of live attenuated and formaldehyde-inactivated Venezuelan equine encephalitis virus vaccines against aerosol challenge in hamsters.. J Clin Microbiol 1984 Mar;19(3):429-31.
    pmc: PMC271080pubmed: 6715512doi: 10.1128/jcm.19.3.429-431.1984google scholar: lookup
  8. Pratt WD, Gibbs P, Pitt ML, Schmaljohn AL. Use of telemetry to assess vaccine-induced protection against parenteral and aerosol infections of Venezuelan equine encephalitis virus in non-human primates.. Vaccine 1998 May-Jun;16(9-10):1056-64.
    pubmed: 9682359doi: 10.1016/s0264-410x(97)00192-8google scholar: lookup
  9. Markland W, McQuaid TJ, Jain J, Kwong AD. Broad-spectrum antiviral activity of the IMP dehydrogenase inhibitor VX-497: a comparison with ribavirin and demonstration of antiviral additivity with alpha interferon.. Antimicrob Agents Chemother 2000 Apr;44(4):859-66.
    pmc: PMC89783pubmed: 10722482doi: 10.1128/aac.44.4.859-866.2000google scholar: lookup
  10. Julander JG, Bowen RA, Rao JR, Day C, Shafer K, Smee DF, Morrey JD, Chu CK. Treatment of Venezuelan equine encephalitis virus infection with (-)-carbodine.. Antiviral Res 2008 Dec;80(3):309-15.
    pmc: PMC2612642pubmed: 18675850doi: 10.1016/j.antiviral.2008.07.002google scholar: lookup
  11. Selvam P, Vijayalakshimi P, Smee DF, Gowen BB, Julander JG, Day CW, Barnard DL. Novel 3-sulphonamido-quinazolin-4(3H)-one derivatives: microwave-assisted synthesis and evaluation of antiviral activities against respiratory and biodefense viruses.. Antivir Chem Chemother 2007;18(5):301-5.
    pubmed: 18046963doi: 10.1177/095632020701800506google scholar: lookup
  12. Kehn-Hall K, Narayanan A, Lundberg L, Sampey G, Pinkham C, Guendel I, Van Duyne R, Senina S, Schultz KL, Stavale E, Aman MJ, Bailey C, Kashanchi F. Modulation of GSK-3β activity in Venezuelan equine encephalitis virus infection.. PLoS One 2012;7(4):e34761.
    pmc: PMC3319612pubmed: 22496857doi: 10.1371/journal.pone.0034761google scholar: lookup
  13. Fine DL, Roberts BA, Teehee ML, Terpening SJ, Kelly CL, Raetz JL, Baker DC, Powers AM, Bowen RA. Venezuelan equine encephalitis virus vaccine candidate (V3526) safety, immunogenicity and efficacy in horses.. Vaccine 2007 Feb 26;25(10):1868-76.
    pubmed: 17240002doi: 10.1016/j.vaccine.2006.10.030google scholar: lookup
  14. Pratt WD, Davis NL, Johnston RE, Smith JF. Genetically engineered, live attenuated vaccines for Venezuelan equine encephalitis: testing in animal models.. Vaccine 2003 Sep 8;21(25-26):3854-62.
    pubmed: 12922119doi: 10.1016/s0264-410x(03)00328-1google scholar: lookup
  15. Pauwels R, Andries K, Desmyter J, Schols D, Kukla MJ, Breslin HJ, Raeymaeckers A, Van Gelder J, Woestenborghs R, Heykants J. Potent and selective inhibition of HIV-1 replication in vitro by a novel series of TIBO derivatives.. Nature 1990 Feb 1;343(6257):470-4.
    pubmed: 1689015doi: 10.1038/343470a0google scholar: lookup
  16. Julander JG, Skirpstunas R, Siddharthan V, Shafer K, Hoopes JD, Smee DF, Morrey JD. C3H/HeN mouse model for the evaluation of antiviral agents for the treatment of Venezuelan equine encephalitis virus infection.. Antiviral Res 2008 Jun;78(3):230-41.
    pmc: PMC2396595pubmed: 18313150doi: 10.1016/j.antiviral.2008.01.007google scholar: lookup
  17. Sawicki DL, Sawicki SG. Short-lived minus-strand polymerase for Semliki Forest virus.. J Virol 1980 Apr;34(1):108-18.
    pmc: PMC288676pubmed: 6768898doi: 10.1128/jvi.34.1.108-118.1980google scholar: lookup
  18. HARDY FM. The growth of Venezuelan equine encephalomyelitis virus in various tissue cultures.. Am J Hyg 1959 Jul;70(1):21-7.
    pubmed: 13670164doi: 10.1093/oxfordjournals.aje.a120061google scholar: lookup
  19. Scherer WF, Ellsworth CA, Ventura AK. Studies of viral virulence. II. Growth and adsorption curves of virulent and attenuated strains of Venezuelan encephalitis virus in cultured cells.. Am J Pathol 1971 Feb;62(2):211-9.
    pmc: PMC2047534pubmed: 5099760
  20. Gomez de Cedrón M, Ehsani N, Mikkola ML, García JA, Kääriäinen L. RNA helicase activity of Semliki Forest virus replicase protein NSP2.. FEBS Lett 1999 Apr 1;448(1):19-22.
    pubmed: 10217401doi: 10.1016/s0014-5793(99)00321-xgoogle scholar: lookup
  21. Rikkonen M, Peränen J, Kääriäinen L. ATPase and GTPase activities associated with Semliki Forest virus nonstructural protein nsP2.. J Virol 1994 Sep;68(9):5804-10.
    pmc: PMC236984pubmed: 8057461doi: 10.1128/jvi.68.9.5804-5810.1994google scholar: lookup
  22. Gorbalenya AE, Koonin EV. Helicases - Amino-Acid-Sequence Comparisons and Structure-Function-Relationships. Current Opinion in Structural Biology 3: 419–429.
  23. Lulla V, Merits A, Sarin P, Kääriäinen L, Keränen S, Ahola T. Identification of mutations causing temperature-sensitive defects in Semliki Forest virus RNA synthesis.. J Virol 2006 Mar;80(6):3108-11.
    pmc: PMC1395444pubmed: 16501123doi: 10.1128/jvi.80.6.3108-3111.2006google scholar: lookup
  24. Hardy WR, Hahn YS, de Groot RJ, Strauss EG, Strauss JH. Synthesis and processing of the nonstructural polyproteins of several temperature-sensitive mutants of Sindbis virus.. Virology 1990 Jul;177(1):199-208.
    pubmed: 2141204doi: 10.1016/0042-6822(90)90473-5google scholar: lookup
  25. Russo AT, White MA, Watowich SJ. The crystal structure of the Venezuelan equine encephalitis alphavirus nsP2 protease.. Structure 2006 Sep;14(9):1449-58.
    pubmed: 16962975doi: 10.1016/j.str.2006.07.010google scholar: lookup
  26. Zhang D, Tözsér J, Waugh DS. Molecular cloning, overproduction, purification and biochemical characterization of the p39 nsp2 protease domains encoded by three alphaviruses.. Protein Expr Purif 2009 Mar;64(1):89-97.
    pmc: PMC2706578pubmed: 19013248doi: 10.1016/j.pep.2008.10.013google scholar: lookup
  27. Sawicki DL, Perri S, Polo JM, Sawicki SG. Role for nsP2 proteins in the cessation of alphavirus minus-strand synthesis by host cells.. J Virol 2006 Jan;80(1):360-71.
    pmc: PMC1317508pubmed: 16352561doi: 10.1128/jvi.80.1.360-371.2006google scholar: lookup
  28. Akhrymuk I, Kulemzin SV, Frolova EI. Evasion of the innate immune response: the Old World alphavirus nsP2 protein induces rapid degradation of Rpb1, a catalytic subunit of RNA polymerase II.. J Virol 2012 Jul;86(13):7180-91.
    pmc: PMC3416352pubmed: 22514352doi: 10.1128/jvi.00541-12google scholar: lookup
  29. Kehn-Hall K, Narayanan A, Lundberg L, Sampey G, Pinkham C, Guendel I, Van Duyne R, Senina S, Schultz KL, Stavale E, Aman MJ, Bailey C, Kashanchi F. Modulation of GSK-3β activity in Venezuelan equine encephalitis virus infection.. PLoS One 2012;7(4):e34761.
    pmc: PMC3319612pubmed: 22496857doi: 10.1371/journal.pone.0034761google scholar: lookup
  30. Huggins JW, Jahrling PB, Rill W, Linden CD. Characterization of the binding of the TC-83 strain of Venezuelan Equine encephalomyelitis virus to BW-J-M, a mouse macrophage-like cell line.. J Gen Virol 1983 Jan;64 (Pt 1):149-57.
    pubmed: 6337232doi: 10.1099/0022-1317-64-1-149google scholar: lookup
  31. Bernard KA, Klimstra WB, Johnston RE. Mutations in the E2 glycoprotein of Venezuelan equine encephalitis virus confer heparan sulfate interaction, low morbidity, and rapid clearance from blood of mice.. Virology 2000 Oct 10;276(1):93-103.
    pubmed: 11021998doi: 10.1006/viro.2000.0546google scholar: lookup
  32. Chung DH, Jonsson CB, Maddox C, McKellip SN, Moore BP, Heil M, White EL, Ananthan S, Li Q, Feng S, Rasmussen L. HTS-driven discovery of new chemotypes with West Nile Virus inhibitory activity.. Molecules 2010 Mar 12;15(3):1690-704.
    pmc: PMC4839297pubmed: 20336008doi: 10.3390/molecules15031690google scholar: lookup
  33. Zhang JH, Chung TD, Oldenburg KR. A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays.. J Biomol Screen 1999;4(2):67-73.
    pubmed: 10838414doi: 10.1177/108705719900400206google scholar: lookup
  34. Dower K, Rubins KH, Hensley LE, Connor JH. Development of Vaccinia reporter viruses for rapid, high content analysis of viral function at all stages of gene expression.. Antiviral Res 2011 Jul;91(1):72-80.
    pmc: PMC3177160pubmed: 21569797doi: 10.1016/j.antiviral.2011.04.014google scholar: lookup
  35. Whitlow ZW, Connor JH, Lyles DS. Preferential translation of vesicular stomatitis virus mRNAs is conferred by transcription from the viral genome.. J Virol 2006 Dec;80(23):11733-42.
    pmc: PMC1642595pubmed: 17005665doi: 10.1128/jvi.00971-06google scholar: lookup
  36. Chung DH, Moore BP, Matharu DS, Golden JE, Maddox C, Rasmussen L, Sosa MI, Ananthan S, White EL, Jia F, Jonsson CB, Severson WE. A cell based high-throughput screening approach for the discovery of new inhibitors of respiratory syncytial virus.. Virol J 2013 Jan 10;10:19.
    pmc: PMC3621174pubmed: 23302182doi: 10.1186/1743-422x-10-19google scholar: lookup
  37. Gupta CK, Leszczynski J, Gupta RK, Siber GR. Stabilization of respiratory syncytial virus (RSV) against thermal inactivation and freeze-thaw cycles for development and control of RSV vaccines and immune globulin.. Vaccine 1996 Oct;14(15):1417-20.
    pubmed: 8994316doi: 10.1016/s0264-410x(96)00096-5google scholar: lookup

Citations

This article has been cited 28 times.
  1. Skidmore AM, Bradfute SB. The life cycle of the alphaviruses: From an antiviral perspective. Antiviral Res 2023 Jan;209:105476.
    doi: 10.1016/j.antiviral.2022.105476pubmed: 36436722google scholar: lookup
  2. Kafai NM, Diamond MS, Fox JM. Distinct Cellular Tropism and Immune Responses to Alphavirus Infection. Annu Rev Immunol 2022 Apr 26;40:615-649.
    doi: 10.1146/annurev-immunol-101220-014952pubmed: 35134315google scholar: lookup
  3. Huang PT, Saul S, Einav S, Asquith CRM. Optimization of 4-Anilinoquinolines as Dengue Virus Inhibitors. Molecules 2021 Dec 3;26(23).
    doi: 10.3390/molecules26237338pubmed: 34885921google scholar: lookup
  4. Haese NN, May NA, Taft-Benz S, Moukha-Chafiq O, Madadi N, Zhang S, Karyakarte SD, Rodzinak KJ, Nguyen TH, Denton M, Streblow AD, Towers NA, Rasmussen L, Bostwick RJ, Maddry JA, Ananthan S, Augelli-Szafran CE, Suto MJ, Sanders W, Moorman N, DeFilippis V, Heise MT, Pathak AK, Streblow DN, Morrison TE. Identification of Quinolinones as Antivirals against Venezuelan Equine Encephalitis Virus. Antimicrob Agents Chemother 2021 Aug 17;65(9):e0024421.
    doi: 10.1128/AAC.00244-21pubmed: 34152810google scholar: lookup
  5. Shechter S, Thomas DR, Jans DA. Application of In Silico and HTS Approaches to Identify Nuclear Import Inhibitors for Venezuelan Equine Encephalitis Virus Capsid Protein: A Case Study. Front Chem 2020;8:573121.
    doi: 10.3389/fchem.2020.573121pubmed: 33505952google scholar: lookup
  6. Lee J, Parvathareddy J, Yang D, Bansal S, O'Connell K, Golden JE, Jonsson CB. Emergence and Magnitude of ML336 Resistance in Venezuelan Equine Encephalitis Virus Depend on the Microenvironment. J Virol 2020 Oct 27;94(22).
    doi: 10.1128/JVI.00317-20pubmed: 32878897google scholar: lookup
  7. Skidmore AM, Adcock RS, Jonsson CB, Golden JE, Chung DH. Benzamidine ML336 inhibits plus and minus strand RNA synthesis of Venezuelan equine encephalitis virus without affecting host RNA production. Antiviral Res 2020 Feb;174:104674.
    doi: 10.1016/j.antiviral.2019.104674pubmed: 31816348google scholar: lookup
  8. Jonsson CB, Cao X, Lee J, Gabbard JD, Chu YK, Fitzpatrick EA, Julander J, Chung DH, Stabenow J, Golden JE. Efficacy of a ML336 derivative against Venezuelan and eastern equine encephalitis viruses. Antiviral Res 2019 Jul;167:25-34.
    doi: 10.1016/j.antiviral.2019.04.004pubmed: 30970271google scholar: lookup
  9. Jaffett VA, Nerurkar A, Cao X, Guzei IA, Golden JE. Telescoped synthesis of C3-functionalized (E)-arylamidines using Ugi-Mumm and regiospecific quinazolinone rearrangements. Org Biomol Chem 2019 Mar 20;17(12):3118-3128.
    doi: 10.1039/c9ob00073apubmed: 30730519google scholar: lookup
  10. LaBauve AE, Rinker TE, Noureddine A, Serda RE, Howe JY, Sherman MB, Rasley A, Brinker CJ, Sasaki DY, Negrete OA. Lipid-Coated Mesoporous Silica Nanoparticles for the Delivery of the ML336 Antiviral to Inhibit Encephalitic Alphavirus Infection. Sci Rep 2018 Sep 18;8(1):13990.
    doi: 10.1038/s41598-018-32033-wpubmed: 30228359google scholar: lookup
  11. Urakova N, Kuznetsova V, Crossman DK, Sokratian A, Guthrie DB, Kolykhalov AA, Lockwood MA, Natchus MG, Crowley MR, Painter GR, Frolova EI, Frolov I. β-d-N(4)-Hydroxycytidine Is a Potent Anti-alphavirus Compound That Induces a High Level of Mutations in the Viral Genome. J Virol 2018 Feb 1;92(3).
    doi: 10.1128/JVI.01965-17pubmed: 29167335google scholar: lookup
  12. Ching KC, F P Ng L, Chai CLL. A compendium of small molecule direct-acting and host-targeting inhibitors as therapies against alphaviruses. J Antimicrob Chemother 2017 Nov 1;72(11):2973-2989.
    doi: 10.1093/jac/dkx224pubmed: 28981632google scholar: lookup
  13. Chung DH, Golden JE, Adcock RS, Schroeder CE, Chu YK, Sotsky JB, Cramer DE, Chilton PM, Song C, Anantpadma M, Davey RA, Prodhan AI, Yin X, Zhang X. Discovery of a Broad-Spectrum Antiviral Compound That Inhibits Pyrimidine Biosynthesis and Establishes a Type 1 Interferon-Independent Antiviral State. Antimicrob Agents Chemother 2016 Aug;60(8):4552-62.
    doi: 10.1128/AAC.00282-16pubmed: 27185801google scholar: lookup
  14. Anantpadma M, Kouznetsova J, Wang H, Huang R, Kolokoltsov A, Guha R, Lindstrom AR, Shtanko O, Simeonov A, Maloney DJ, Maury W, LaCount DJ, Jadhav A, Davey RA. Large-Scale Screening and Identification of Novel Ebola Virus and Marburg Virus Entry Inhibitors. Antimicrob Agents Chemother 2016 Aug;60(8):4471-81.
    doi: 10.1128/AAC.00543-16pubmed: 27161622google scholar: lookup
  15. Hu X, Compton JR, Leary DH, Olson MA, Lee MS, Cheung J, Ye W, Ferrer M, Southall N, Jadhav A, Morazzani EM, Glass PJ, Marugan J, Legler PM. Kinetic, Mutational, and Structural Studies of the Venezuelan Equine Encephalitis Virus Nonstructural Protein 2 Cysteine Protease. Biochemistry 2016 May 31;55(21):3007-19.
    doi: 10.1021/acs.biochem.5b00992pubmed: 27030368google scholar: lookup
  16. Keck F, Ataey P, Amaya M, Bailey C, Narayanan A. Phosphorylation of Single Stranded RNA Virus Proteins and Potential for Novel Therapeutic Strategies. Viruses 2015 Oct 12;7(10):5257-73.
    doi: 10.3390/v7102872pubmed: 26473910google scholar: lookup
  17. Li C, Guillén J, Rabah N, Blanjoie A, Debart F, Vasseur JJ, Canard B, Decroly E, Coutard B. mRNA Capping by Venezuelan Equine Encephalitis Virus nsP1: Functional Characterization and Implications for Antiviral Research. J Virol 2015 Aug;89(16):8292-303.
    doi: 10.1128/JVI.00599-15pubmed: 26041283google scholar: lookup
  18. Matharu DS, Flaherty DP, Simpson DS, Schroeder CE, Chung D, Yan D, Noah JW, Jonsson CB, White EL, Aubé J, Plemper RK, Severson WE, Golden JE. Optimization of potent and selective quinazolinediones: inhibitors of respiratory syncytial virus that block RNA-dependent RNA-polymerase complex activity. J Med Chem 2014 Dec 26;57(24):10314-28.
    doi: 10.1021/jm500902xpubmed: 25399509google scholar: lookup
  19. Schroeder CE, Yao T, Sotsky J, Smith RA, Roy S, Chu YK, Guo H, Tower NA, Noah JW, McKellip S, Sosa M, Rasmussen L, Smith LH, White EL, Aubé J, Jonsson CB, Chung D, Golden JE. Development of (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-5-nitro-N-phenylbenzamide, ML336: Novel 2-amidinophenylbenzamides as potent inhibitors of venezuelan equine encephalitis virus. J Med Chem 2014 Oct 23;57(20):8608-21.
    doi: 10.1021/jm501203vpubmed: 25244572google scholar: lookup
  20. Chung D, Otsuka Y, Kim E, Ullah S, Kennedy NM, Ngo KH, Kang J, Tomlison R, Alejandro B, Barnaby K, Miller J, Shumate J, Teo CF, Tan YB, Chew CS, Kang C, Luo D, Scampavia L, Spicer TP, Bannister TD. Discovery of a potent anti-Zika virus benzamide series targeting the viral protein NS4B. bioRxiv 2025 Oct 9;.
    doi: 10.1101/2025.10.09.681361pubmed: 41279169google scholar: lookup
  21. Williams EP, Xue Y, Yang D, Lee J, Kim E, Ponce-Flores A, Zalduondo L, Cao X, Chung D, Golden JE, Meibohm B, Fitzpatrick EA, Weaver SC, Jonsson CB. Broad spectrum antiviral BDGR-164 provides protection against lethal neurotropic alphavirus infection in mice. Antiviral Res 2025 Aug;240:106206.
    doi: 10.1016/j.antiviral.2025.106206pubmed: 40456491google scholar: lookup
  22. Wang S, Mahalingam S, Merits A. Alphavirus nsP2: A Multifunctional Regulator of Viral Replication and Promising Target for Anti-Alphavirus Therapies. Rev Med Virol 2025 Mar;35(2):e70030.
    doi: 10.1002/rmv.70030pubmed: 40064592google scholar: lookup
  23. Williams EP, Xue Y, Vogel P, Yang D, Ponce-Flores A, Li X, Ogorek TJ, Saini M, Iulek J, Ruiz FX, Arnold E, Golden JE, Meibohm B, Jonsson CB. The antiviral BDGR-49 provides protection from lethal, neurotropic Venezuelan equine encephalitis virus intranasal infection in mice. J Virol 2025 Mar 18;99(3):e0167924.
    doi: 10.1128/jvi.01679-24pubmed: 39936916google scholar: lookup
  24. VanderGiessen M, de Jager C, Leighton J, Xie H, Theus M, Johnson E, Kehn-Hall K. Neurological manifestations of encephalitic alphaviruses, traumatic brain injuries, and organophosphorus nerve agent exposure. Front Neurosci 2024;18:1514940.
    doi: 10.3389/fnins.2024.1514940pubmed: 39734493google scholar: lookup
  25. Merten EM, Sears JD, Leisner TM, Hardy PB, Ghoshal A, Hossain MA, Asressu KH, Brown PJ, Tse EG, Stashko MA, Li K, Norris-Drouin JL, Herring LE, Mordant AL, Webb TS, Mills CA, Barker NK, Streblow ZJ, Perveen S, Arrowsmith CH, Couñago RM, Arnold JJ, Cameron CE, Streblow DN, Moorman NJ, Heise MT, Willson TM, Popov KI, Pearce KH. Identification of a cell-active chikungunya virus nsP2 protease inhibitor using a covalent fragment-based screening approach. Proc Natl Acad Sci U S A 2024 Oct 15;121(42):e2409166121.
    doi: 10.1073/pnas.2409166121pubmed: 39388272google scholar: lookup
  26. Rasmussen L, Sanders S, Sosa M, McKellip S, Nebane NM, Martinez-Gzegozewska Y, Reece A, Ruiz P, Manuvakhova A, Zhai L, Warren B, Curry A, Zeng Q, Bostwick JR, Vinson PN. A high-throughput response to the SARS-CoV-2 pandemic. SLAS Discov 2024 Jul;29(5):100160.
    doi: 10.1016/j.slasd.2024.100160pubmed: 38761981google scholar: lookup
  27. Marques RE, Shimizu JF, Nogueira ML, Vasilakis N. Current challenges in the discovery of treatments against Mayaro fever. Expert Opin Ther Targets 2024 May;28(5):345-356.
    doi: 10.1080/14728222.2024.2351504pubmed: 38714500google scholar: lookup
  28. Cao X, Yang D, Parvathareddy J, Chu YK, Kim EJ, Fitz-Henley JN, Li X, Lukka PB, Parmar KR, Temrikar ZH, Dhole P, Adcock RS, Gabbard J, Bansal S, Lee J, Zalduondo L, Hayes E, Stabenow J, Meibohm B, Fitzpatrick EA, Bailey K, Campos RK, Julander JG, Rossi SL, Chung D, Jonsson CB, Golden JE. Efficacy of a brain-penetrant antiviral in lethal Venezuelan and eastern equine encephalitis mouse models. Sci Transl Med 2023 Apr 12;15(691):eabl9344.
    doi: 10.1126/scitranslmed.abl9344pubmed: 37043558google scholar: lookup
Subscribe to our newsletter
Join 99,928 horse owners like you who receive our email updates on horse health and nutrition.