The manuscript entitled "Leishmanicidal Potential of Hardwickiic Acid Isolated From Croton sylvaticus" is a scientific piece of work published in Frontiers in Pharmacology. The comments on the abstract, title and references are as below

1● The title of this manuscript is informative and relevant and clearly indicates the objective and key findings of the research work 2● The abstract of this paper is concisely elaborate on the epidemiology, hypothesis, objective, and outcome of the work. Overall informative.

3● The references are relevant, mostly recent but few references are 10-20 years old but cited correctly.

Comments on introduction/background

The introduction part is written in a sequential manner and interesting in reading. It starts with the epidemiological data followed by available treatment options and their drawback but need more emphasis on the mode of action of available drugs. The hypothesis part is also elaborative and connects the objective. Overall

1. It clearly elaborates on what is already known about this topic.
2. The research question is clearly outlined, and
3. The research question justified given what is already known about the topic

But minor corrections needed 1. Give more emphasis on the mode of action of available drugs 2. Author should mention that why they selected TR as drug target instead of all other available drug targets 3. As in natural condition a single molecule have different isomeric form, how the author will justify the acive presence of a specific isomer

Comments on methodology

The methodology part is defined and measured appropriately and prove the reliability of the method. However, the details are not much enough to replicate this study. The minor corrections are mentioned below

Comments on data and results

I find the work interesting and potentially impactful but have major concerns over the quality of the manuscript. The results and discussion are too superficial. These parts, in particular, need to be more insightful to create a coherent picture of the entire work.

1. As the main objective of this study was to search for effective replacement of available treatment options but the obtained IC50 was found to be higher as compare to control drug Amphotericin B. How the authors will justify it?
2. They checked the IC50 using promastigote form of the parasite but this form is present in sandfly. How this compound will be effective against the amastigote forms of parasite? Please explain?
3. The authors checked the IC50 using amphotericin B as a reference drug. As per my opinion, as the molecular weight of HA is 316g/mol, it should be proposed as an oral drug and should be compared with the miltefosine.
4. Authors used the dug for either cell line or parasite treatment but mentioned the volume of drug they should mention the stock concentration from which they took a certain volume of drug
5. As the authors went for the Trypanothione docking, they must perform ROS analysis to validate the in silico outcome. They can use the H2DCFDA probe for the same.
6. All the figures are low in resolution. they must submit the figures in TIF format with 600DPI resolution
7. The author should check the IC50 value using amastigote/axenic amastigotes of leishmania parasite.

Comments on discussion and conclusions

The authors isolated and tested the compound and compare the results with positive control. They evaluated the selectivity index too. The author used docking to propose the mechanism of action.

One important limitation of the manuscript is to compare compounds using mass concentration instead of using molar concentration. The authors must compare compounds with different molar mass using molar concentration, therefore whole discussion section must using molar concentration values instead of mass concentration.

The authors must compare compounds with different molar mass using molar concentration, therefore, please, the whole discussion section, and consequently the conclusion must use molar concentration values instead of mass concentration. Please replace all concentrations (IC50 and CC50) in µg/mL units with molar concentrations.

Abstract and Conclusion "SI = 7.84 ± 0.01 µg/mL" - Selectivity index is dimensionless, please erase the unit " µg/mL".