Dear Dr. Alami,

Thank you very much for submitting your manuscript "A Phase-I pharmacokinetic, safety and food effect study on flubentylosin, a novel analog of Tylosin-A having potent anti-Wolbachia activity and antifilarial activity" for consideration at PLOS Neglected Tropical Diseases. As with all papers reviewed by the journal, your manuscript was reviewed by members of the editorial board and by several independent reviewers. In light of the reviews (below this email), we would like to invite the resubmission of a significantly-revised version that takes into account the reviewers' comments.

Authors should consider the comments raised by both reviewers adding more details about Wolbachia in filarioids in the introduction and reorganization of the discussion section (see reviewer 2)

We cannot make any decision about publication until we have seen the revised manuscript and your response to the reviewers' comments. Your revised manuscript is also likely to be sent to reviewers for further evaluation.

When you are ready to resubmit, please upload the following:

[1] A letter containing a detailed list of your responses to the review comments and a description of the changes you have made in the manuscript. Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out.

[2] Two versions of the revised manuscript: one with either highlights or tracked changes denoting where the text has been changed; the other a clean version (uploaded as the manuscript file).

Important additional instructions are given below your reviewer comments.

Please prepare and submit your revised manuscript within 60 days. If you anticipate any delay, please let us know the expected resubmission date by replying to this email. Please note that revised manuscripts received after the 60-day due date may require evaluation and peer review similar to newly submitted manuscripts.

Thank you again for your submission. We hope that our editorial process has been constructive so far, and we welcome your feedback at any time. Please don't hesitate to contact us if you have any questions or comments.

Sincerely,

Domenico Otranto

Deputy Editor

PLOS Neglected Tropical Diseases

Domenico Otranto

Deputy Editor

PLOS Neglected Tropical Diseases

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Authors should consider the comments raised by both reviewers adding more details about Wolbachia in filarioids in the introduction and reorganization of the discussion section (see reviewer 2)

Reviewer's Responses to Questions

Key Review Criteria Required for Acceptance?

As you describe the new analyses required for acceptance, please consider the following:

Methods

-Are the objectives of the study clearly articulated with a clear testable hypothesis stated?

-Is the study design appropriate to address the stated objectives?

-Is the population clearly described and appropriate for the hypothesis being tested?

-Is the sample size sufficient to ensure adequate power to address the hypothesis being tested?

-Were correct statistical analysis used to support conclusions?

-Are there concerns about ethical or regulatory requirements being met?

Reviewer #1: The manuscript describes the results of a phase-1 safety trial with flubentylosin, which targets Wolbachia in filarial nematodes.

General comments. The manuscript is well written; the premise is correct (the need to evaluate safety profile of a potentially important drug for the control of filarial disease); the Methods are described clearly and are adequate;

Reviewer #2: (No Response)

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Results

-Does the analysis presented match the analysis plan?

-Are the results clearly and completely presented?

-Are the figures (Tables, Images) of sufficient quality for clarity?

Reviewer #1: the Results are presented with excellent graphical support; the Discussion could put a bit more emphasis on the previous studies cited (the development of the drug/modifications to increase bioavailability following oral administration; results obtained on Wolbachia depletion in the murine model). Also a nice way to remind readers of the importance of the past A-WOL project).

This Reviewer recommends publication following very minor revision.

Abstract. Seventy-eight healthy subjects: replace with “healthy adults”.

Reviewer #2: (No Response)

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Conclusions

-Are the conclusions supported by the data presented?

-Are the limitations of analysis clearly described?

-Do the authors discuss how these data can be helpful to advance our understanding of the topic under study?

-Is public health relevance addressed?

Reviewer #1: the Discussion could put a bit more emphasis on the previous studies cited (the development of the drug/modifications to increase bioavailability following oral administration; results obtained on Wolbachia depletion in the murine model). Also a nice way to remind readers of the importance of the past A-WOL project).

Reviewer #2: (No Response)

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Editorial and Data Presentation Modifications?

Use this section for editorial suggestions as well as relatively minor modifications of existing data that would enhance clarity. If the only modifications needed are minor and/or editorial, you may wish to recommend “Minor Revision” or “Accept”.

Reviewer #1: This Reviewer recommends publication following very minor revision.

Abstract. Seventy-eight healthy subjects: replace with “healthy adults”.

Reviewer #2: (No Response)

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Summary and General Comments

Use this section to provide overall comments, discuss strengths/weaknesses of the study, novelty, significance, general execution and scholarship. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. If requesting major revision, please articulate the new experiments that are needed.

Reviewer #1: The manuscript describes the results of a phase-1 safety trial with flubentylosin, which targets Wolbachia in filarial nematodes.

General comments. The manuscript is well written; the premise is correct (the need to evaluate safety profile of a potentially important drug for the control of filarial disease); the Methods are described clearly and are adequate; the Results are presented with excellent graphical support; the Discussion could put a bit more emphasis on the previous studies cited (the development of the drug/modifications to increase bioavailability following oral administration; results obtained on Wolbachia depletion in the murine model). Also a nice way to remind readers of the importance of the past A-WOL project).

T

Reviewer #2: The manuscript PNTD-D-22-00302, entitled “A Phase-I pharmacokinetic, safety and food effect 1 study on flubentylosin, a novel analog 2 of Tylosin-A having potent anti-Wolbachia activity and antifilarial activity” is a Phase-I pharmacokinetic, safety and food-effect study on single and multiple ascending doses of the macrolide antibacterial drug, flubentylosin (ABBV-4083), targeting filarial endosymbiont, Wolbachia for the effective control of filariasis and onchocerciasis.

Here in this article, authors are assessing the safety and pharmacokinetics of flubentylosin in healthy subjects, to identify a dose and dosing regimen for the future studies. In general, the introduction section should include the role of Wolbachia in filarids and its imunopathological effects in filariasis. Here you are targeting Wolbachia for the treatment and hence more importance should be given for this endosymbiont. Discussion has to be rewritten. Avoid the repetition of results in discussion and discuss your results in detail. References has to be uniformly formatted. Please follow the authors guidelines of the journal while formatting. Please recheck the text carefully for grammatical and formatting mistakes. Please see the attached PDF of the manuscript for my detailed comments for the authors.

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Reviewer #1: Yes: LAURA HELEN KRAMER

Reviewer #2: Yes: Ranju Manoj

Figure Files:

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Attachments

Attachment

Submitted filename: Reviewer comments_Plos.doc

Attachment

Submitted filename: PNTD-D-22-00302_Review.pdf

Dear Alami,

We are pleased to inform you that your manuscript 'A Phase-I pharmacokinetic, safety and food-effect study on flubentylosin, a novel analog of Tylosin-A having potent anti-Wolbachia and antifilarial activity' has been provisionally accepted for publication in PLOS Neglected Tropical Diseases.

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 Neglected Tropical Diseases.

Best regards,

Cinzia Cantacessi

Section Editor

PLOS Neglected Tropical Diseases

Domenico Otranto

Academic Editor

PLOS Neglected Tropical Diseases

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Reviewer's Responses to Questions

Key Review Criteria Required for Acceptance?

As you describe the new analyses required for acceptance, please consider the following:

Methods

-Are the objectives of the study clearly articulated with a clear testable hypothesis stated?

-Is the study design appropriate to address the stated objectives?

-Is the population clearly described and appropriate for the hypothesis being tested?

-Is the sample size sufficient to ensure adequate power to address the hypothesis being tested?

-Were correct statistical analysis used to support conclusions?

-Are there concerns about ethical or regulatory requirements being met?

Reviewer #2: The objectives of the study was clearly articulated with testable hypothesis.

The design was appropriate and have addressed all the objectives

The sample size was adequate and the statistical analysis was appropriate

All ethical requirements were met in the study.

**********

Results

-Does the analysis presented match the analysis plan?

-Are the results clearly and completely presented?

-Are the figures (Tables, Images) of sufficient quality for clarity?

Reviewer #2: The results presented matched with the plan of study

Authors were successful in presenting the results, clearly and concisely

The quality of figures and tables were good

**********

Conclusions

-Are the conclusions supported by the data presented?

-Are the limitations of analysis clearly described?

-Do the authors discuss how these data can be helpful to advance our understanding of the topic under study?

-Is public health relevance addressed?

Reviewer #2: Conclusions were supported by the presented results and the limitations were clearly mentioned. Authors have discussed the significance of the data and its public health significance.

**********

Editorial and Data Presentation Modifications?

Use this section for editorial suggestions as well as relatively minor modifications of existing data that would enhance clarity. If the only modifications needed are minor and/or editorial, you may wish to recommend “Minor Revision” or “Accept”.

Reviewer #2: Accept the article with minor changes

**********

Summary and General Comments

Use this section to provide overall comments, discuss strengths/weaknesses of the study, novelty, significance, general execution and scholarship. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. If requesting major revision, please articulate the new experiments that are needed.

Reviewer #2: I am pleased to inform that the R1 has considerably improved the paper. Authors have carefully addressed the reviewer’s comments. Hence in my opinion this paper can be accepted for publication in PLOS Neglected Tropical Diseases. Please see the pdf for some minor corrections.

**********

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 #2: No

Attachments

Attachment

Submitted filename: PNTD-D-22-00302_R1_reviewer comments.pdf

Attachment

Submitted filename: Reviewer comments R1_PLOS.doc