https://orcid.org/0000-0002-2593-2571
Peer Review History
| Original SubmissionSeptember 5, 2025 |
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PPATHOGENS-D-25-02218 Repurposing of a Clinical Protease Inhibitor Enables Oral, In Vivo Broad-Spectrum Inhibition of Enteroviruses PLOS Pathogens Dear Dr. Zhang, Thank you for submitting your manuscript to PLOS Pathogens. After careful consideration, we feel that it has merit but does not fully meet PLOS Pathogens's publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Please submit your revised manuscript by Jan 31 2026 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plospathogens@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/ppathogens/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. Please include the following items when submitting your revised manuscript: * A rebuttal letter that responds to each point raised by the editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. This file does not need to include responses to any formatting updates and technical items listed in the 'Journal Requirements' section below. * A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. * An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, competing interests statement, or data availability statement, please make these updates within the submission form at the time of resubmission. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. We look forward to receiving your revised manuscript. Kind regards, George A. Belov, PhD Academic Editor PLOS Pathogens Alexander Gorbalenya Section Editor PLOS Pathogens Sumita Bhaduri-McIntosh Editor-in-Chief PLOS Pathogens orcid.org/0000-0003-2946-9497 Michael Malim Editor-in-Chief PLOS Pathogens orcid.org/0000-0002-7699-2064 Additional Editor Comments : - Please refer to picornavirus (virus) taxa, including species according to ICTV rules on nomenclature - Acknowledge key similarities between enterovirus 3Cpro and coronavirus Mpro (which also is known as 3C-like protease, 3CLpro) that are critical for the background of this study and discussion of its results. The manuscript presentation, including the Title, Abstract, Intro and Discussion and Reference list, must be improved to reveal gain of knowledge compared to the updated background. Below are two reviews to relevant literature https://doi.org/10.1016/j.antiviral.2010.12.007https://doi.org/10.1007/BF02174046 Journal Requirements: Note: If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. Reviewers' Comments: Reviewer's Responses to Questions Part I - Summary Please use this section to discuss strengths/weaknesses of study, novelty/significance, general execution and scholarship. Reviewer #1: This manuscript presents a well-executed and highly relevant study that addresses the urgent need for antiviral therapies against enteroviruses. The authors convincingly demonstrate that bofutrelvir, originally developed as a SARS-CoV-2 Mpro inhibitor, potently inhibits enteroviral 3C protease activity and exhibits broad-spectrum antiviral efficacy against multiple enteroviruses, including EV71, Coxsackievirus A16, and Echovirus 11. The combination of enzymatic, cellular, and in vivo studies, supported by structural elucidation of the inhibitor–protease complex, provides a comprehensive understanding of the mechanism of action and underscores the therapeutic potential of bofutrelvir. The demonstrated reduction of viral loads and alleviation of disease symptoms in infected neonatal mice further enhance the translational significance of the work. Reviewer #2: In this manuscript, the authors screened a library of 104 protease inhibitors to search for new inhibitors of EV71 3C protease. They found bofutrelvir, a clinical stage peptidomimetic developed for SARS-CoV2 Mpro, has potent antiviral activity against multiple enteroviruses in cell culture. Crystal structure of bofutrelvir binding to EV71 3C protein was resolved, supporting a mechanism of covalently inhibiting the protease active site. Bofutrelvir demonstrated in vivo efficacy against EV71 infection in neonatal mice through intraperitoneal injection or oral administration, and against CA16 infection through oral administration. The data are convincing, and the experiments are well performed. The discovery of bofutrelvir as a broad-spectrum enterovirus 3C inhibitor provides the foundation for repurposing this compound for antivirals against enterovirus diseases. ********** Part II – Major Issues: Key Experiments Required for Acceptance Please use this section to detail the key new experiments or modifications of existing experiments that should be absolutely required to validate study conclusions. Generally, there should be no more than 3 such required experiments or major modifications for a "Major Revision" recommendation. If more than 3 experiments are necessary to validate the study conclusions, then you are encouraged to recommend "Reject". Reviewer #1: bofutrelvir was previously reported by authors as enterovirus 3CLpro inhibitors. This needs to be updated and clearly spelled out in the introduction. Reviewer #2: What is the survival curve of the neonatal mice after EV71 and CA16 infection? does bofutrelvir provide any protection against mortality? The author showed that the efficacy of bofutrelvir in cell culture is comparable to or even better than that of rupintrivir, the known enterovirus 3C inhibitor. Comparison beyond this point is lacking. Rupintrivir has failed clinical trials. It is not clear from the manuscript whether bofutrelvir has any advantage over rupintrivir. The author can use their own experimental data or extrapolate from published dataset for such comparisons, which may include: 1. How does the bofutrelvir-3C structure compare to rupintrivir-3C structure? The author briefly mentions that the two structures are similar. A more detailed comparison should be included. 2. what is the in vivo efficacy and dose range of rupintrivir in the neonatal mouse model, or other in vivo model? 3. how does the bioavailability of bofutrelvir and rupintrivir compare? ********** Part III – Minor Issues: Editorial and Data Presentation Modifications Please use this section for editorial suggestions as well as relatively minor modifications of existing data that would enhance clarity. Reviewer #1: "Several antiviral agents targeting different stages of the EVs life cycle have been investigated" Comment: reviews of EV-A71 and EV-D68 antivirals should be updated Acta Pharm. Sin. B. 2022, 12, 1542-1566. Lab Invest. 2024 Feb;104(2):100298. ACS Infect. Dis. 2020, 6, 1572-1586. Viruses 2025, 17(9), 1178 "One possible explanation for this discrepancy is that these compounds preferentially bind to 139 endogenous caspase family proteases within the host cells, which share similar catalytic cysteine 140 residues" Comment: another possibility might be the lack of cellular permeability due to the negative charge of the aspartic acid. Discussion session should be updated with the translational potential of aldehyde containing drugs. Reviewer #2: Fig.2B what is the estimated EC50 according to the TCID50 assay? How does it compare to the qPCR assay in Fig.2A? Why were clinical scores in Fig. 4 a lot less than Fig.6? Line 282: “with strong antiviral effects seen in peripheral tissues such as the spinal cord and hind limb muscles”. Spinal cord is not a peripheral tissue. ********** PLOS authors have the option to publish the peer review history of their article (what does this mean? ). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy . Reviewer #1: No Reviewer #2: No [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] Figure resubmission: Reproducibility: To enhance the reproducibility of your results, we recommend that authors of applicable studies deposit laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. Additionally, PLOS ONE offers an option to publish peer-reviewed clinical study protocols. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols |
| Revision 1 |
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PPATHOGENS-D-25-02218R1 Repurposing of a Clinical Protease Inhibitor Enables Oral, In Vivo Broad-Spectrum Inhibition of Enteroviruses PLOS Pathogens Dear Dr. Zhang, Thank you for submitting your manuscript to PLOS Pathogens. After careful consideration, we feel that it has merit but does not fully meet PLOS Pathogens's publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process (see below signature line). Please submit your revised manuscript by Apr 06 2026 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plospathogens@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/ppathogens/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. Please include the following items when submitting your revised manuscript: * A letter that responds to each point raised by the editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. This file does not need to include responses to any formatting updates and technical items listed in the 'Journal Requirements' section below. * A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. * An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, competing interests statement, or data availability statement, please make these updates within the submission form at the time of resubmission. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. We look forward to receiving your revised manuscript. Kind regards, George A. Belov, PhD Academic Editor PLOS Pathogens Alexander Gorbalenya Section Editor PLOS Pathogens Sumita Bhaduri-McIntosh Editor-in-Chief PLOS Pathogens orcid.org/0000-0003-2946-9497 Michael Malim Editor-in-Chief PLOS Pathogens orcid.org/0000-0002-7699-2064 Additional Editor Comments: Please fully address the editorial suggestions from the previous review. Specifically: 1) Improve accuracy and detail of the title, e.g. " A clinical orally-available SARS-CoV-2 Mpro inhibitor blocks replication of multiple enteroviruses in cell culture and controls enterovirus A71 infection in animal models" ; 2) Restructure Abstract and place the known link between 3Cpro and Mpro BEFORE your screening results; 3) Author Summary. Replace COVID-19 with SARS-CoV-2. Specify the known link between picornavirus 3Cpro and coronavirus Mpro that made success of your research feasible; 4) Revisit the logic and gain of knowledge of your study in the Introduction and Discussion; 5) Please check with ICTV web-site if names of species enterovirus A-D and rhinovirus A-C remain actual and, if necessary, update the text accordingly; 6) Please reverse edits of the first sentence of the Introduction and keep plural form for enteroviruses and rhinoviruses (there are many of those); 7) Consider acknowledging the Picornaviridae Study Group for the Picornaviridae taxonomy and foundational research on enterovirus/picornavirus molecular biology; 8) Highlight changes to the Ref list; Reviewers' Comments: [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] Figure resubmission: Reproducibility: To enhance the reproducibility of your results, we recommend that authors of applicable studies deposit laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. Additionally, PLOS ONE offers an option to publish peer-reviewed clinical study protocols. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols |
| Revision 2 |
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Dear Dr. Zhang, We are pleased to inform you that your manuscript 'A Clinical SARS-CoV-2 Mpro Inhibitor Blocks Replication of Multiple Enteroviruses and Confers Oral In Vivo Protection in Animal Models' has been provisionally accepted for publication in PLOS Pathogens. Before your manuscript can be formally accepted you will need to complete some formatting changes, which you will receive in a follow up email. A member of our team will be in touch with a set of requests. Please note that your manuscript will not be scheduled for publication until you have made the required changes, so a swift response is appreciated. IMPORTANT: The editorial review process is now complete. PLOS will only permit corrections to spelling, formatting or significant scientific errors from this point onwards. Requests for major changes, or any which affect the scientific understanding of your work, will cause delays to the publication date of your manuscript. Should you, your institution's press office or the journal office choose to press release your paper, you will automatically be opted out of early publication. We ask that you notify us now if you or your institution is planning to press release the article. All press must be co-ordinated with PLOS. Thank you again for supporting Open Access publishing; we are looking forward to publishing your work in PLOS Pathogens. Best regards, George A. Belov, PhD Academic Editor PLOS Pathogens Alexander Gorbalenya Section Editor PLOS Pathogens Sumita Bhaduri-McIntosh Editor-in-Chief PLOS Pathogens orcid.org/0000-0003-2946-9497 Michael Malim Editor-in-Chief PLOS Pathogens orcid.org/0000-0002-7699-2064 *********************************************************** Reviewer Comments (if any, and for reference): |
| Formally Accepted |
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Dear Dr. Zhang, We are delighted to inform you that your manuscript, "A Clinical SARS-CoV-2 Mpro Inhibitor Blocks Replication of Multiple Enteroviruses and Confers Oral In Vivo Protection in Animal Models," has been formally accepted for publication in PLOS Pathogens. We have now passed your article onto the PLOS Production Department who will complete the rest of the pre-publication process. All authors will receive a confirmation email upon publication. The corresponding author will soon be receiving a typeset proof for review, to ensure errors have not been introduced during production. Please review the PDF proof of your manuscript carefully, as this is the last chance to correct any scientific or type-setting errors. Please note that major changes, or those which affect the scientific understanding of the work, will likely cause delays to the publication date of your manuscript. Note: Proofs for Front Matter articles (Pearls, Reviews, Opinions, etc...) are generated on a different schedule and may not be made available as quickly. Soon after your final files are uploaded, the early version of your manuscript, if you opted to have an early version of your article, will be published online. The date of the early version will be your article's publication date. The final article will be published to the same URL, and all versions of the paper will be accessible to readers. For Research Articles, you will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing. Thank you again for supporting open-access publishing; we are looking forward to publishing your work in PLOS Pathogens. Best regards, Sumita Bhaduri-McIntosh Editor-in-Chief PLOS Pathogens orcid.org/0000-0003-2946-9497 Michael Malim Editor-in-Chief PLOS Pathogens orcid.org/0000-0002-7699-2064 |
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