PONE-D-20-04266

KD-64 – a new selective A2A adenosine receptor antagonist has anti-inflammatory activity but contrary to the non-selective antagonist – caffeine does not reduce diet-induced obesity in mice

PLOS ONE

Dear Dr Kotańska,

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

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Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: General Comments:

According to the World Health Organization (WHO), obesity has reached epidemic proportions globally. Obesity causes low-grade chronic inflammation which may lead to a number of chronic diseases. Additionally, A2 adenosine receptors play an important role in the regulation of the inflammatory process and glucose homeostasis.

Although caffeine, a non-selective A2A adenosine receptor antagonist, has been extensively studied by some research groups, the anti-inflammatory and anti-obesity mechanisms of some selective A2A adenosine receptor antagonists are few studied.

This paper reports the anti-inflammatory and anti-obesity activities of KD-64, a new selective A2A adenosine receptor antagonist, by using several current techniques and animal models.

The paper is original, well written and organized, and within the scope of Plos One.

Specific Comments

Title

The title should be concise. Authors should consider to reduce it.

Introduction

In page 5 – lines 106 and 107, it is referred that “…These compounds have similar structure to KD-64 – ligand we have used in the presented study…”.

KD-64 IUPAC chemical name must be included and the similarities between the chemical structures of KD-64 and the other referred compounds should be described.

Is KD-64 a caffeine analogue?

Experimental Design / Results and Discussion

Concerning the three animal models, more details must be included, namely:

• The age of the animals at the beginning of the experiences

• Considering that gender is a confounding variable, wouldn’t be better to use only one gender in all groups/models of the study? (Usually, male mice provide more pure and accurate responses…)

• The concentration of caffeine used was much higher than the concentration of KD-64. Why? (This may explain, at least partially, the reported caffeine pro-inflammatory effect).

• Standard diet composition should be provided in a table (together with the high fat-diet composition)

Reviewer #2: Dear Kotanska and colleagues,

I have some minor questions regarding your article:

- line 73-75 "the role of adenosine receptor signaling in the development and progressiona of numerous diseases has been emphasized for years" needs a reference.

- line 146: "in all experiments ketoprofenm caffeine or KD-64 were administered as suspensions in 1% Tween 80" in what solution? PBS? NaCL?

- line 258: how did you measure "spontanous activity"?

- figure 4 C in yy axis please add the wavelenght the absorbance was measured

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Reviewer #1: No

Reviewer #2: No

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KD-64 – a new selective A2A adenosine receptor antagonist has anti-inflammatory activity but contrary to the non-selective antagonist – caffeine does not reduce diet-induced obesity in mice

PONE-D-20-04266R1

Dear Dr. Kotańska,

We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements.

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With kind regards,

Marco Aurélio Gouveia Alves

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: All comments have been addressed

Reviewer #3: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

Reviewer #3: Yes

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #2: Yes

Reviewer #3: Yes

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4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #2: Yes

Reviewer #3: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #2: Yes

Reviewer #3: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #2: Dear authors,

Thank you for revising the manuscript accordingly with reviewers instructions. It is now suitable for publication.

Reviewer #3: (No Response)

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7. 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

Reviewer #3: No