Rapid Discovery of Antimicrobial and Antimalarial Agents from Natural Product Fragments

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

In this study, the authors described that the benefits of using a pure natural product library for fragment-based drug discovery. Below are some comments to revise before publication:

1.      The title needs revision as it currently resembles a review article. Please specify the novelty of your research. Additionally, clarify the type of natural product you are focusing on.

2.      Abstract should summarize all important finding. What about fraction 30-36? Please explain these in abstract part.

3.      What were the criteria for selecting the natural product in this study? Why were these plants and microbe chosen? An introduction to these products should be explained.

4.      Full name of all abbreviation should be first mentioned. BCG, SA, MRSA

5.      Table 2: Please check the table caption

Comments on the Quality of English Language

Minor grammatical corrections can enhance the readability of the manuscript.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript authored by Han and co-workers presents a well-designed and meticulously executed study. The research is of high quality and holds significant potential for publication in the Journal of Separation. However, the manuscript contains numerous typographical errors and instances of missing abbreviations for acronyms that need to be addressed to enhance clarity and readability. Below are some examples:

• Line 18: Please provide the full forms of BCG, SA, and MRSA when they are first mentioned in the text.
• Line 79: Include the full name of DMSO when it is first referenced.
• Line 96: Clarify the abbreviation TCMs to ensure it is understandable to the readers.
• Lines 80-82: The manuscript should provide complete information about the instruments used, specifically NMR, LC-MS, UHPLC, and HRMS. Include the model numbers, manufacturers, and the full address of the suppliers.

In addition to these corrections, I recommend a thorough proofread of the manuscript to rectify any remaining typographical errors and ensure that all abbreviations are defined upon their initial usage.

Overall, the manuscript will be well-suited for publication in the Journal of Separation.

Author Response

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The research on "Fragment-based Drug Discovery from a Natural Product Library" offers a compelling exploration of utilizing natural product fragments for drug discovery, emphasizing the advantages of natural product diversity and inherent bioactivity over synthetic libraries. The study presents a robust methodology for constructing a high-quality natural product fragment library and demonstrates the biological potential of the isolated fragments, particularly against antimicrobial targets. Notably, the research introduces a native mass spectrometry technique for rapid identification of non-competitive fragments, enhancing the efficiency of fragment-based drug discovery. However, while the study's approach is innovative and promising, there are some limitations such as the reproducibility of extracting active compounds from natural sources and the relatively narrow focus on specific protein targets. These aspects may affect the general applicability of the findings to a broader range of drug discovery projects.

1.  How does the study address the potential variability and reproducibility issues in extracting active compounds from diverse natural sources, given the inherent complexity and variability of natural products?

2. Why did the study choose to focus on a limited number of malarial proteins, and how might the findings be generalized to other therapeutic targets or disease areas?

3. What measures were taken to ensure the chemical diversity of the natural product fragment library, and how does this diversity compare to that of traditional synthetic fragment libraries?

4. Can you elaborate on the limitations and potential challenges of using native mass spectrometry for fragment identification, particularly in terms of sensitivity and specificity in complex biological mixtures?

Comments on the Quality of English Language

The quality of English language in the manuscript is generally good; however, it requires editing due to a high similarity percentage, indicating potential issues with originality.

Author Response

Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

Comments and Suggestions for Authors

The author has made significant efforts to enhance the content in response to reviewer feedback. This study presents an innovative and effective approach to constructing a natural product fragment library, highlighting the inherent diversity and bioactivity advantages over synthetic libraries. The author addressed variability and reproducibility issues in extracting active compounds from natural sources through a single-step extraction process optimized for cost and time while maintaining high quality. The selected and validated extraction methods, such as ethanol and n-butanol extractions, ensure consistent and reproducible results. Furthermore, the research extends the application of native mass spectrometry in identifying non-competitive fragments, providing a solid foundation for developing more potent antimalarial inhibitors through fragment linking or merging. These improvements not only ensure consistency and reproducibility but also open potential avenues for new drug discovery projects. The positive feedback from reviewers reflects the author's commitment and substantial effort in perfecting the manuscript.

Comments on the Quality of English Language

The quality of the English language in this manuscript is generally good. The text is clear, coherent, and well-structured, effectively conveying the scientific concepts and methodologies