Bisbenzimide compounds inhibit the replication of prototype and pandemic potential poxviruses

Jerzy Samolej; Diogo Correa Mendonca; Nicole Upfold; Marion McElwee; Mariann Landsberger; Artur Yakimovich; Arvind H Patel; Blair L Strang; Jason Mercer

doi:10.1128/spectrum.04072-23

Bisbenzimide compounds inhibit the replication of prototype and pandemic potential poxviruses

Jerzy Samolej, Diogo Correa Mendonca, Nicole Upfold, Marion McElwee, Mariann Landsberger, Artur Yakimovich, Arvind H Patel, Blair L Strang^*, Jason Mercer^*

^*Corresponding author for this work

Biosciences

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Abstract

We previously identified the bisbenzimide Hoechst 33342 (H42) as a potent multi-stage inhibitor of the prototypic poxvirus, the vaccinia virus (VACV), and several parapoxviruses. A recent report showed that novel bisbenzimide compounds similar in structure to H42 could prevent human cytomegalovirus replication. Here, we assessed whether these compounds could also serve as poxvirus inhibitors. Using virological assays, we show that these bisbenzimide compounds inhibit VACV spread, plaque formation, and the production of infectious progeny VACV with relatively low cell toxicity. Further analysis of the VACV lifecycle indicated that the effective bisbenzimide compounds had little impact on VACV early gene expression but inhibited VACV late gene expression and truncated the formation of VACV replication sites. Additionally, we found that bisbenzimide compounds, including H42, can inhibit both monkeypox and a VACV mutant resistant to the widely used anti-poxvirus drug TPOXX (Tecovirimat). Therefore, the tested bisbenzimide compounds were inhibitors of both prototypic and pandemic potential poxviruses and could be developed for use in situations where anti-poxvirus drug resistance may occur. Additionally, these data suggest that bisbenzimide compounds may serve as broad-activity antiviral compounds, targeting diverse DNA viruses such as poxviruses and betaherpesviruses.IMPORTANCEThe 2022 mpox (monkeypox) outbreak served as a stark reminder that due to the cessation of smallpox vaccination over 40 years ago, most of the human population remains susceptible to poxvirus infection. With only two antivirals approved for the treatment of smallpox infection in humans, the need for additional anti-poxvirus compounds is evident. Having shown that the bisbenzimide H33342 is a potent inhibitor of poxvirus gene expression and DNA replication, here we extend these findings to include a set of novel bisbenzimide compounds that show anti-viral activity against mpox and a drug-resistant prototype poxvirus mutant. These results suggest that further development of bisbenzimides for the treatment of pandemic potential poxviruses is warranted.

Original language	English
Article number	e0407223
Journal	Microbiology spectrum
Volume	12
Issue number	4
Early online date	20 Feb 2024
DOIs	https://doi.org/10.1128/spectrum.04072-23
Publication status	Published - 2 Apr 2024

Bibliographical note

ACKNOWLEDGMENTS
We thank Andy Merritt (LifeArc) for the generous provision of reagents and his support of the project throughout. We thank Dennis Hruby and Douglas Grosenbach (SIGA Technologies, Inc.) for providing TPOXX.
B.L.S. was supported by St George’s, University of London. J.M. was supported by the Medical Research Council (MC_PC_19029) and the BBSRC-funded mpox rapid response grant BB/X011607/1.
The mpox virus work was carried out by CRUSH: Antiviral Drug Screening and Resistance Hub at the MRC-University of Glasgow Centre for Virus Research (CVR).
Funding for this research was supported by the BBSRC-funded mpox rapid response grant BB/X011607/1, LifeArc COVID-19 award, and the MRC core award MC_UU_00034/9 (A.H.P.). This work was partially funded by the Center for Advanced Systems Understanding (CASUS), which is financed by Germany’s Federal Ministry of Education and Research (BMBF) and by the Saxon Ministry for Science, Culture, and Tourism (SMWK) with tax funds on the basis of the budget approved by the Saxon State Parliament (A.Y.).
J.S. analyzed and curated the data, designed the methodology, supervised the study, wrote the original draft, and reviewed and edited the manuscript. D.C.M. and N.U. conceptualized the study, designed the methodology, performed the investigation, analyzed and curated the data, and reviewed and edited the manuscript. M.Mc.E. and M.L. performed the investigation. A.Y. performed the investigation and reviewed and edited the manuscript. A.H.P. supervised the study, reviewed and edited the manuscript, and acquired funding. B.L.S. conceptualized the study, designed the methodology, wrote the original draft, and reviewed and edited the manuscript. J.P.M. conceptualized the study, designed the methodology, wrote the original draft, reviewed and edited the manuscript, supervised the study, administrated the project, and acquired funding.

Keywords

Humans
Poxviridae
Bisbenzimidazole/metabolism
Smallpox
Pandemics
Vaccinia virus/genetics

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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SamoleJ2024BisbenzimideFinal published version, 2.43 MBLicence: Creative Commons: Attribution (CC BY)

Cite this

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title = "Bisbenzimide compounds inhibit the replication of prototype and pandemic potential poxviruses",

abstract = "We previously identified the bisbenzimide Hoechst 33342 (H42) as a potent multi-stage inhibitor of the prototypic poxvirus, the vaccinia virus (VACV), and several parapoxviruses. A recent report showed that novel bisbenzimide compounds similar in structure to H42 could prevent human cytomegalovirus replication. Here, we assessed whether these compounds could also serve as poxvirus inhibitors. Using virological assays, we show that these bisbenzimide compounds inhibit VACV spread, plaque formation, and the production of infectious progeny VACV with relatively low cell toxicity. Further analysis of the VACV lifecycle indicated that the effective bisbenzimide compounds had little impact on VACV early gene expression but inhibited VACV late gene expression and truncated the formation of VACV replication sites. Additionally, we found that bisbenzimide compounds, including H42, can inhibit both monkeypox and a VACV mutant resistant to the widely used anti-poxvirus drug TPOXX (Tecovirimat). Therefore, the tested bisbenzimide compounds were inhibitors of both prototypic and pandemic potential poxviruses and could be developed for use in situations where anti-poxvirus drug resistance may occur. Additionally, these data suggest that bisbenzimide compounds may serve as broad-activity antiviral compounds, targeting diverse DNA viruses such as poxviruses and betaherpesviruses.IMPORTANCEThe 2022 mpox (monkeypox) outbreak served as a stark reminder that due to the cessation of smallpox vaccination over 40 years ago, most of the human population remains susceptible to poxvirus infection. With only two antivirals approved for the treatment of smallpox infection in humans, the need for additional anti-poxvirus compounds is evident. Having shown that the bisbenzimide H33342 is a potent inhibitor of poxvirus gene expression and DNA replication, here we extend these findings to include a set of novel bisbenzimide compounds that show anti-viral activity against mpox and a drug-resistant prototype poxvirus mutant. These results suggest that further development of bisbenzimides for the treatment of pandemic potential poxviruses is warranted.",

keywords = "Humans, Poxviridae, Bisbenzimidazole/metabolism, Smallpox, Pandemics, Vaccinia virus/genetics",

author = "Jerzy Samolej and Mendonca, {Diogo Correa} and Nicole Upfold and Marion McElwee and Mariann Landsberger and Artur Yakimovich and Patel, {Arvind H} and Strang, {Blair L} and Jason Mercer",

note = "ACKNOWLEDGMENTS We thank Andy Merritt (LifeArc) for the generous provision of reagents and his support of the project throughout. We thank Dennis Hruby and Douglas Grosenbach (SIGA Technologies, Inc.) for providing TPOXX. B.L.S. was supported by St George{\textquoteright}s, University of London. J.M. was supported by the Medical Research Council (MC_PC_19029) and the BBSRC-funded mpox rapid response grant BB/X011607/1. The mpox virus work was carried out by CRUSH: Antiviral Drug Screening and Resistance Hub at the MRC-University of Glasgow Centre for Virus Research (CVR). Funding for this research was supported by the BBSRC-funded mpox rapid response grant BB/X011607/1, LifeArc COVID-19 award, and the MRC core award MC_UU_00034/9 (A.H.P.). This work was partially funded by the Center for Advanced Systems Understanding (CASUS), which is financed by Germany{\textquoteright}s Federal Ministry of Education and Research (BMBF) and by the Saxon Ministry for Science, Culture, and Tourism (SMWK) with tax funds on the basis of the budget approved by the Saxon State Parliament (A.Y.). J.S. analyzed and curated the data, designed the methodology, supervised the study, wrote the original draft, and reviewed and edited the manuscript. D.C.M. and N.U. conceptualized the study, designed the methodology, performed the investigation, analyzed and curated the data, and reviewed and edited the manuscript. M.Mc.E. and M.L. performed the investigation. A.Y. performed the investigation and reviewed and edited the manuscript. A.H.P. supervised the study, reviewed and edited the manuscript, and acquired funding. B.L.S. conceptualized the study, designed the methodology, wrote the original draft, and reviewed and edited the manuscript. J.P.M. conceptualized the study, designed the methodology, wrote the original draft, reviewed and edited the manuscript, supervised the study, administrated the project, and acquired funding.",

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doi = "10.1128/spectrum.04072-23",

language = "English",

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journal = "Microbiology spectrum",

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T1 - Bisbenzimide compounds inhibit the replication of prototype and pandemic potential poxviruses

AU - Samolej, Jerzy

AU - Mendonca, Diogo Correa

AU - Upfold, Nicole

AU - McElwee, Marion

AU - Landsberger, Mariann

AU - Yakimovich, Artur

AU - Patel, Arvind H

AU - Strang, Blair L

AU - Mercer, Jason

N1 - ACKNOWLEDGMENTS We thank Andy Merritt (LifeArc) for the generous provision of reagents and his support of the project throughout. We thank Dennis Hruby and Douglas Grosenbach (SIGA Technologies, Inc.) for providing TPOXX. B.L.S. was supported by St George’s, University of London. J.M. was supported by the Medical Research Council (MC_PC_19029) and the BBSRC-funded mpox rapid response grant BB/X011607/1. The mpox virus work was carried out by CRUSH: Antiviral Drug Screening and Resistance Hub at the MRC-University of Glasgow Centre for Virus Research (CVR). Funding for this research was supported by the BBSRC-funded mpox rapid response grant BB/X011607/1, LifeArc COVID-19 award, and the MRC core award MC_UU_00034/9 (A.H.P.). This work was partially funded by the Center for Advanced Systems Understanding (CASUS), which is financed by Germany’s Federal Ministry of Education and Research (BMBF) and by the Saxon Ministry for Science, Culture, and Tourism (SMWK) with tax funds on the basis of the budget approved by the Saxon State Parliament (A.Y.). J.S. analyzed and curated the data, designed the methodology, supervised the study, wrote the original draft, and reviewed and edited the manuscript. D.C.M. and N.U. conceptualized the study, designed the methodology, performed the investigation, analyzed and curated the data, and reviewed and edited the manuscript. M.Mc.E. and M.L. performed the investigation. A.Y. performed the investigation and reviewed and edited the manuscript. A.H.P. supervised the study, reviewed and edited the manuscript, and acquired funding. B.L.S. conceptualized the study, designed the methodology, wrote the original draft, and reviewed and edited the manuscript. J.P.M. conceptualized the study, designed the methodology, wrote the original draft, reviewed and edited the manuscript, supervised the study, administrated the project, and acquired funding.

PY - 2024/4/2

Y1 - 2024/4/2

N2 - We previously identified the bisbenzimide Hoechst 33342 (H42) as a potent multi-stage inhibitor of the prototypic poxvirus, the vaccinia virus (VACV), and several parapoxviruses. A recent report showed that novel bisbenzimide compounds similar in structure to H42 could prevent human cytomegalovirus replication. Here, we assessed whether these compounds could also serve as poxvirus inhibitors. Using virological assays, we show that these bisbenzimide compounds inhibit VACV spread, plaque formation, and the production of infectious progeny VACV with relatively low cell toxicity. Further analysis of the VACV lifecycle indicated that the effective bisbenzimide compounds had little impact on VACV early gene expression but inhibited VACV late gene expression and truncated the formation of VACV replication sites. Additionally, we found that bisbenzimide compounds, including H42, can inhibit both monkeypox and a VACV mutant resistant to the widely used anti-poxvirus drug TPOXX (Tecovirimat). Therefore, the tested bisbenzimide compounds were inhibitors of both prototypic and pandemic potential poxviruses and could be developed for use in situations where anti-poxvirus drug resistance may occur. Additionally, these data suggest that bisbenzimide compounds may serve as broad-activity antiviral compounds, targeting diverse DNA viruses such as poxviruses and betaherpesviruses.IMPORTANCEThe 2022 mpox (monkeypox) outbreak served as a stark reminder that due to the cessation of smallpox vaccination over 40 years ago, most of the human population remains susceptible to poxvirus infection. With only two antivirals approved for the treatment of smallpox infection in humans, the need for additional anti-poxvirus compounds is evident. Having shown that the bisbenzimide H33342 is a potent inhibitor of poxvirus gene expression and DNA replication, here we extend these findings to include a set of novel bisbenzimide compounds that show anti-viral activity against mpox and a drug-resistant prototype poxvirus mutant. These results suggest that further development of bisbenzimides for the treatment of pandemic potential poxviruses is warranted.

AB - We previously identified the bisbenzimide Hoechst 33342 (H42) as a potent multi-stage inhibitor of the prototypic poxvirus, the vaccinia virus (VACV), and several parapoxviruses. A recent report showed that novel bisbenzimide compounds similar in structure to H42 could prevent human cytomegalovirus replication. Here, we assessed whether these compounds could also serve as poxvirus inhibitors. Using virological assays, we show that these bisbenzimide compounds inhibit VACV spread, plaque formation, and the production of infectious progeny VACV with relatively low cell toxicity. Further analysis of the VACV lifecycle indicated that the effective bisbenzimide compounds had little impact on VACV early gene expression but inhibited VACV late gene expression and truncated the formation of VACV replication sites. Additionally, we found that bisbenzimide compounds, including H42, can inhibit both monkeypox and a VACV mutant resistant to the widely used anti-poxvirus drug TPOXX (Tecovirimat). Therefore, the tested bisbenzimide compounds were inhibitors of both prototypic and pandemic potential poxviruses and could be developed for use in situations where anti-poxvirus drug resistance may occur. Additionally, these data suggest that bisbenzimide compounds may serve as broad-activity antiviral compounds, targeting diverse DNA viruses such as poxviruses and betaherpesviruses.IMPORTANCEThe 2022 mpox (monkeypox) outbreak served as a stark reminder that due to the cessation of smallpox vaccination over 40 years ago, most of the human population remains susceptible to poxvirus infection. With only two antivirals approved for the treatment of smallpox infection in humans, the need for additional anti-poxvirus compounds is evident. Having shown that the bisbenzimide H33342 is a potent inhibitor of poxvirus gene expression and DNA replication, here we extend these findings to include a set of novel bisbenzimide compounds that show anti-viral activity against mpox and a drug-resistant prototype poxvirus mutant. These results suggest that further development of bisbenzimides for the treatment of pandemic potential poxviruses is warranted.

KW - Humans

KW - Poxviridae

KW - Bisbenzimidazole/metabolism

KW - Smallpox

KW - Pandemics

KW - Vaccinia virus/genetics

U2 - 10.1128/spectrum.04072-23

DO - 10.1128/spectrum.04072-23

M3 - Article

C2 - 38376353

SN - 2165-0497

VL - 12

JO - Microbiology spectrum

JF - Microbiology spectrum

IS - 4

M1 - e0407223

ER -

Bisbenzimide compounds inhibit the replication of prototype and pandemic potential poxviruses

Abstract

Bibliographical note

Keywords

UN SDGs

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