Ginkgetin: A Promising Multitarget Agent for Diverse Diseases

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

1. Detailed Manuscript Summary
The pharmacological landscape of the C-C linked biflavonoid ginkgetin is presented in this review. The authors are successful in bridging the gap between its pleiotropic effects in neurology, inflammatory diseases, and oncology and its natural origins. The current body of knowledge is greatly enhanced by the addition of contemporary pathways like cGAS-STING. 

2. Extensive Critical Evaluation and Commentary for Authors

A. Introduction and Abstract: Determining the Scientific Justification

-Although the abstract accurately notes GK's "broad pharmacological activities," the compound's distinct biochemical niche is not yet clearly defined. What distinguishes GK from other biflavonoids (like amentoflavone) should be made clear by the authors.

-The burden of chronic illness is well-established in the Introduction, but there is no justification for this particular review. In contrast to more widely researched flavonoids like quercetin, the authors must explain why GK in particular advantages such a thorough review at this time. What special therapeutic window does GK provide that isn't available to others?
  
  B. Chemical Architecture and SAR Analysis (Section 2.1)
The authors should incorporate a more thorough discussion of the Structure-Activity Relationship (SAR) in order to present a professional-grade analysis. In particular, I would like to see an examination of how the apigenin units' methoxy and hydroxyl locations determine their affinity for the Nrf2 or STING pathways. Does the dimeric "twist" angle serve as a scaffold for the functional groups or is it necessary for binding? 

C. Mechanistic Hierarchy and Signaling Complexity 
Although the manuscript offers an incredible number of pathways (MAPK, PI3K/Akt, etc.), it does not include a "hierarchy of effects." It is frequently difficult to distinguish between secondary downstream effects and fundamental mechanisms (direct binding/inhibition). Differentiating between these is essential to comprehending GK's actual mode of activity.    
 
D. Therapeutic potential of ginkgetin across disease systems (Section 4)
  The sections on Cancer and Neurological Disorders rely heavily on preclinical data. To provide a more clinical perspective, the authors should add a dedicated subsection or table critiquing the limitations of these models .Specifically, they must address the use of high, non-physiological concentrations in vitro (e.g., >20 μM) which rarely translate to human systemic levels.

  E.Pharmacokinetic considerations and druggability challenges  
  Although "low oral bioavailability" is a standard observation, a high-impact study ought to include solutions. I recommend that the writers investigate and talk about new approaches to get around GK's lipophilicity, like co-delivery methods, prodrug development, or nano-formulations.

3.Conclusion section :  I encourage the authors to propose a specific "Scientific Roadmap" for the next five years. What are the definitive, must-have experiments (e.g., standardized toxicology, Phase 0 micro-dosing, or specific target-engagement assays) required to move GK from "preclinical promise" to a viable clinical trial candidate?

Conclusion of the report  : The manuscript is a strong candidate for publication if it transitions from a descriptive report to a critical analytical piece. Correcting the terminology (GK as a lead agent rather than a target) and addressing the pharmacokinetic hurdles will significantly improve the manuscript's standing in the field.

Author Response

Dear Editors and Reviewers:

Thank you for allowing me to revise our manuscript (ID 4190452). We thank the reviewers for their constructive feedback, which has contributed to the clarity and rigor of our work. We processed each comment carefully and responded to the reviewer's suggestions in detail below. Revisions in the manuscript are highlighted for your reference.

Comments: Detailed Manuscript Summary

The pharmacological landscape of the C-C-linked biflavonoid ginkgetin is presented in this review. The authors are successful in bridging the gap between its pleiotropic effects in neurology, inflammatory diseases, and oncology and its natural origins. The current body of knowledge is greatly enhanced by the addition of contemporary pathways like cGAS-STING.

Response: Thank you for your positive and encouraging comments on our manuscript. We sincerely appreciate your recognition of our work, especially your acknowledgment regarding our efforts to bridge the pleiotropic effects of ginkgetin with its natural origins and to incorporate contemporary pathways such as cGAS-STING. Your supportive summary is a great motivation for us.

Below, we have provided a point-by-point response to each of your specific comments and have revised the manuscript accordingly. All changes are highlighted in the manuscript for your convenience.

 

Comments:  Extensive Critical Evaluation and Commentary for Authors

1. Introduction and Abstract: Determining the Scientific Justification

Comment 1:  Although the abstract accurately notes GK's "broad pharmacological activities," the compound's distinct biochemical niche is not yet clearly defined. What distinguishes GK from other biflavonoids (like amentoflavone) should be made clear by the authors.

Response 1:  We sincerely thank the reviewer for this insightful and constructive comment. Highlighting the unique value of GK compared to other biflavonoids is crucial for positioning our review. In response, we have revised the abstract to clearly state the distinctive characteristics of GK. Specifically, we have added a sentence following the first introductory sentence to emphasize.

 

Comment 2:  The burden of chronic illness is well-established in the Introduction, but there is no justification for this particular review. In contrast to more widely researched flavonoids like quercetin, the authors must explain why GK in particular advantages such a thorough review at this time. What special therapeutic window does GK provide that isn't available to others?

Response 2:   We sincerely thank the reviewer for raising this fundamental point. We agree that a stronger justification for a dedicated review on GK is crucial. In direct response to your comment, we have added a new, dedicated paragraph in the Introduction (see lines 94-112 in the revised manuscript) to explicitly address these questions.

 

Comment 3:  Chemical Architecture and SAR Analysis (Section 2.1)

The authors should incorporate a more thorough discussion of the Structure-Activity Relationship (SAR) in order to present a professional-grade analysis. In particular, I would like to see an examination of how the apigenin units' methoxy and hydroxyl locations determine their affinity for the Nrf2 or STING pathways. Does the dimeric "twist" angle serve as a scaffold for the functional groups or is it necessary for binding?

Response 3:   We thank the reviewer for this insightful suggestion to deepen our SAR analysis. In direct response, we have substantially expanded Section 2.1 (now titled “Chemical structure, physicochemical properties, and structure-activity relationships”)​ to incorporate a professional-grade SAR discussion.

 

Comment 4:  Mechanistic Hierarchy and Signaling Complexity

Although the manuscript offers an incredible number of pathways (MAPK, PI3K/Akt, etc.), it does not include a "hierarchy of effects." It is frequently difficult to distinguish between secondary downstream effects and fundamental mechanisms (direct binding/inhibition). Differentiating between these is essential to comprehending GK's actual mode of activity.  

Response 4:  We are deeply grateful to the reviewer for this critical and insightful comment, which has fundamentally improved the conceptual rigor of our manuscript. We fully agree that establishing a mechanistic hierarchy is essential. In direct and comprehensive response, we have completely restructured the entire "Molecular Mechanisms" chapter (Section 3)​ to introduce and apply a clear hierarchical model.  

 

Comment 5:  Therapeutic potential of ginkgetin across disease systems (Section 4)

The sections on Cancer and Neurological Disorders rely heavily on preclinical data. To provide a more clinical perspective, the authors should add a dedicated subsection or table critiquing the limitations of these models. Specifically, they must address the use of high, non-physiological concentrations in vitro (e.g., >20 μM) which rarely translate to human systemic levels.

Response 5:  Thank you for your valuable suggestion regarding the need for a more critical appraisal of the preclinical evidence. In response, we have added a new subsection "4.1.1. Critical Appraisal: Translational Challenges and Model Limitations in Oncology" (lines 447-472) and "4.4.1. Critical Appraisal: Translational Challenges and Model Limitations in Neurology" (lines 551-576) to address this issue.

 

Comment 6:  Pharmacokinetic considerations and druggability challenges  

Although "low oral bioavailability" is a standard observation, a high-impact study ought to include solutions. I recommend that the writers investigate and talk about new approaches to get around GK's lipophilicity, like co-delivery methods, prodrug development, or nano-formulations.

Response 6:  Thank you for your insightful advice on the pharmacokinetic challenges and the need for specific solutions. In response, we have made major revisions to Section 5, shifting it from a descriptive summary of the challenge to a more comprehensive and solution-oriented discussion.

 

Comment 7:  Conclusion section:  I encourage the authors to propose a specific "Scientific Roadmap" for the next five years. What are the definitive, must-have experiments (e.g., standardized toxicology, Phase 0 micro-dosing, or specific target-engagement assays) required to move GK from "preclinical promise" to a viable clinical trial candidate?

Response 7:  We thank the reviewer for the excellent suggestion to propose a specific "Scientific Roadmap" for the future development of Ginkgetin. In response, we have substantially revised the Conclusion section, which is now titled "7. Conclusion and a Translational Roadmap ", to transform it from a general summary into a forward-looking, strategic blueprint that outlines definitive experiments required to advance GK toward clinical trials. This addition significantly enhances the manuscript's impact by offering concrete guidance for future research and development efforts.

 

Comment 8:  Conclusion of the report : The manuscript is a strong candidate for publication if it transitions from a descriptive report to a critical analytical piece. Correcting the terminology (GK as a lead agent rather than a target) and addressing the pharmacokinetic hurdles will significantly improve the manuscript's standing in the field.

Response 8:  Thank you for your overall evaluation of the manuscript. We have made a comprehensive revision of the manuscript based on your suggestion, moving from a descriptive report to a more critically analytical article. And we corrected terminology use (explicitly describing GK as a "lead" rather than a "target") and substantially expanded the solution section for pharmacokinetic disorders.

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript provides a narrative review of ginkgetin (GK), a biflavonoid derived mainly from Ginkgo biloba, summarizing its chemical characteristics, molecular mechanisms, and therapeutic potential across multiple disease systems, including cancer, metabolic disorders, cardiovascular diseases, neurological disorders, skeletal diseases, and bacterial infections.

The topic is timely and relevant, as natural polyphenolic compounds with multitarget pharmacological properties have attracted increasing attention in drug discovery and pharmacological research. The manuscript is generally well organized and clearly structured, moving logically from chemical characteristics to molecular mechanisms and therapeutic applications.

However, several issues should be addressed before the manuscript can be considered for publication. In particular, the manuscript would benefit from a stronger critical analysis of the literature, improved methodological transparency, and a more cautious interpretation of the therapeutic potential of ginkgetin.

1. Lack of critical analysis of the literature

Although the manuscript summarizes a large number of studies, the discussion remains largely descriptive rather than analytical.

For example, in the section discussing molecular mechanisms (Section 3), several signaling pathways are mentioned (Nrf2/HO-1, NF-κB, MAPK, JAK/STAT, PI3K/Akt, cGAS-STING). However, the manuscript does not clearly distinguish:evidence derived from in vitro experiments, results obtained from animal models, direct mechanistic evidence versus associative observations.

2. Overinterpretation of therapeutic potential

In several sections, the manuscript presents ginkgetin as a promising therapeutic agent for numerous diseases, including cancer, metabolic disorders, and neurodegenerative diseases. However, most of the studies cited are preclinical investigations, and there is currently very limited clinical evidence supporting the therapeutic application of ginkgetin in humans.

3. Absence of summary tables

Given the large amount of information presented, the manuscript would benefit greatly from additional summary tables.

For example, the authors could include: a table summarizing disease models used to evaluate ginkgetin. A table listing molecular targets and signaling pathways affected by ginkgetin. A table summarizing reported pharmacological activities and experimental evidence.

 

Author Response

Dear Editors and Reviewers:

Thank you for giving us the opportunity to revise our manuscript (ID 4190452). We sincerely appreciate the reviewers' insightful comments and valuable suggestions, which have helped us significantly improve the quality of our work. We have carefully addressed all the points raised and have provided point-by-point responses below. The revisions in the manuscript have been highlighted for your convenience.

Comments:  The manuscript provides a narrative review of ginkgetin (GK), a biflavonoid derived mainly from Ginkgo biloba, summarizing its chemical characteristics, molecular mechanisms, and therapeutic potential across multiple disease systems, including cancer, metabolic disorders, cardiovascular diseases, neurological disorders, skeletal diseases, and bacterial infections.

The topic is timely and relevant, as natural polyphenolic compounds with multitarget pharmacological properties have attracted increasing attention in drug discovery and pharmacological research. The manuscript is generally well organized and clearly structured, moving logically from chemical characteristics to molecular mechanisms and therapeutic applications.

However, several issues should be addressed before the manuscript can be considered for publication. In particular, the manuscript would benefit from a stronger critical analysis of the literature, improved methodological transparency, and a more cautious interpretation of the therapeutic potential of ginkgetin.

Response: We sincerely thank the reviewer for the positive assessment of our manuscript's topic, relevance, and overall structure. We are grateful for the reviewer’s time and valuable insights, which are crucial for enhancing the quality of our work. We provide a point-by-point response to each of the specific comments that follow this summary. We have carefully revised the manuscript in accordance with all suggestions. All changes are clearly indicated in the revised manuscript for the reviewer’s convenience.

 

Comment 1:  Lack of critical analysis of the literature

Although the manuscript summarizes a large number of studies, the discussion remains largely descriptive rather than analytical.

For example, in the section discussing molecular mechanisms (Section 3), several signaling pathways are mentioned (Nrf2/HO-1, NF-κB, MAPK, JAK/STAT, PI3K/Akt, cGAS-STING). However, the manuscript does not clearly distinguish: evidence derived from in vitro experiments, results obtained from animal models, direct mechanistic evidence versus associative observations.

Response 1: We thank the reviewer for this crucial comment, which has guided us to significantly enhance the analytical depth and scientific rigor of the manuscript. We agree that distinguishing between levels and types of evidence is fundamental to a critical review. In direct and comprehensive response, we have implemented a systematic revision throughout the manuscript, with particular focus on Section 3 (“Molecular Mechanisms”), to incorporate the requested critical analysis.

 

Comment 2:  Overinterpretation of therapeutic potential

In several sections, the manuscript presents ginkgetin as a promising therapeutic agent for numerous diseases, including cancer, metabolic disorders, and neurodegenerative diseases. However, most of the studies cited are preclinical investigations, and there is currently very limited clinical evidence supporting the therapeutic application of ginkgetin in humans.

Response 2:  We thank the reviewer for this crucial and valid point. We completely agree that the preclinical nature of the existing evidence must be clearly and consistently highlighted to provide a balanced perspective. In response, we have made the revisions throughout the manuscript according to this comment.

 

Comment 3:  Absence of summary tables

Given the large amount of information presented, the manuscript would benefit greatly from additional summary tables.

For example, the authors could include: a table summarizing disease models used to evaluate ginkgetin. A table listing molecular targets and signaling pathways affected by ginkgetin. A table summarizing reported pharmacological activities and experimental evidence.

Response 3:  Thank you for the valuable suggestion to include summary tables. In response, we have added a comprehensive Table 1 at the end of Section 4, which integrates the requested information: (1) disease models used to evaluate GK, (2) molecular targets and signaling pathways affected, and (3) reported pharmacological activities with corresponding experimental evidence. The table is organized by disease area, with columns specifying key findings, experimental models, molecular mechanisms, and representative references.

Reviewer 3 Report

Comments and Suggestions for Authors

The manuscript contains a comprehensive research summary on an interesting natural product, ginkgetin, and its bioactivities. The article is well-written and structured. I think it is appropriate for the scope of the journal and can be published after just a few minor changes.

One thing: is the title really appropriate? I mean the formulation "promising therapeutic target for diverse diseases" - really a TARGET for DISEASES? Bit it might be a matter of taste.

Further {minor) issues:

• The species names originating from Latin should generally be written in italics.
• All the figures should be also referenced within the text.
• There are two figures called "Figure 1" and twp figures called "Figure 2"
• The literature references are not completely uniform in style.

Author Response

Dear Editors and Reviewers:

Thank you for giving us the opportunity to revise our manuscript (ID 4190452). We sincerely appreciate the reviewers' insightful comments and valuable suggestions, which have helped us significantly improve the quality of our work. We have carefully addressed all the points raised and have provided point-by-point responses below. The revisions in the manuscript have been highlighted for your convenience.

Comments: The manuscript contains a comprehensive research summary on an interesting natural product, ginkgetin, and its bioactivities. The article is well-written and structured. I think it is appropriate for the scope of the journal and can be published after just a few minor changes.

One thing: is the title really appropriate? I mean the formulation "promising therapeutic target for diverse diseases" - really a TARGET for DISEASES? Bit it might be a matter of taste.

Further {minor) issues:

The species names originating from Latin should generally be written in italics.

All the figures should be also referenced within the text.

There are two figures called "Figure 1" and twp figures called "Figure 2"

The literature references are not completely uniform in style.

Response: We sincerely thank you for the positive assessment of our manuscript and the valuable suggestions for improvement. Below we provide point-by-point responses to the minor issues raised.

 

Comment 1: "One thing: is the title really appropriate? I mean the formulation 'promising therapeutic target for diverse diseases' - really a TARGET for DISEASES? Bit it might be a matter of taste."

Response 1:  Thank you for this thoughtful observation. We agree that the wording "therapeutic target" could be misleading, as GK is an agent that targets biological pathways rather than being a target itself. To address this, we have revised the title to: "Ginkgetin: A Promising Multitarget Agent for Diverse Diseases". This revised formulation more accurately reflects GK's role as a bioactive compound that modulates multiple molecular targets.

 

Comment 2: "The species names originating from Latin should generally be written in italics."

Response 2:  Thank you for this important stylistic note. We have carefully reviewed the entire manuscript and ensured that all Latin species names (e.g., Ginkgo biloba, Selaginella, Taxus, Araucaria angustifolia, Gaultheria yunnanensis, Escherichia coli, Streptococcus suis) are now correctly italicized throughout the text, including in the main body, figure legends, and references where applicable.

 

Comment 3: "All the figures should be also referenced within the text."

Response 3:  Thank you for this observation. We have carefully reviewed the manuscript and ensured that every figure is explicitly cited in the text at the appropriate location.

 

Comment 4: " There are two figures called 'Figure 1' and two figures called 'Figure 2'."

Response 4: Thank you for catching this numbering error. We have carefully renumbered all figures sequentially throughout the manuscript to ensure uniqueness and consistency. The figures are now correctly numbered as Figure 1, Figure 2, Figure 3, and Figure 4. We have verified that all in-text citations correspond to the correct figure numbers.

 

Comment 5: "The literature references are not completely uniform in style."

Response 5: Thank the reviewer for this attention to detail. We have thoroughly revised the reference list to ensure consistent formatting according to the journal's requirements. All references now follow a uniform style, including consistent use of journal name abbreviations, punctuation, italics for journal titles and volume numbers, and DOI formatting. We have also verified that all in-text citations match the reference list entries.

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

After reviewing the revised manuscript and the point-by-point rebuttal, I am pleased to confirm that all of my concerns have been addressed comprehensively

Reviewer 2 Report

Comments and Suggestions for Authors

Accept in present form