Review Reports

Round 1

Reviewer 1 Report

The work is interesting and well performed. I think it is suitable for publication. However, the manuscript needs minor modifications before publication. The manuscript should be accurately checked for some typing errors (some of them are highlighted in yellow in the attached file)

Comments for author File: Comments.pdf

Author Response

Dear Editor and Reviewer,

Thank you for your letter and for the reviewers’ comments concerning our manuscript entitled “BA-12 inhibits angiogenesis via glutathione metabolism activation” (ID: ijms-563296). Those comments are all valuable and very helpful for revising and improving our paper, as well as the important guiding significance to our researches. We have studied comments carefully and have made correction which we hope meet with approval. Revised portion are marked using “Track Changes” in the paper. The point to point responds to the reviewer’s comments are listed as follows:

Responds to the reviewer’s comments:

Reviewer 1

The work is interesting and well performed. I think it is suitable for publication. However, the manuscript needs minor modifications before publication. The manuscript should be accurately checked for some typing errors (some of them are highlighted in yellow in the attached file)

Response:

Thank you very much for your beneficial comments! We tried to propose one drug discovery strategy based on TCM compatibility theory and combination principle, taking one effective ligustrazine-betulinic acid derivative (BA-12) as an example. Many experiments were carried out to try to express this strategy more objectively. We try our best to replenish these errors according to the Reviewer’s suggestion. The details have been revised using “Track Changes” in the manuscript.

We tried our best to improve the manuscript and made some changes in the manuscript. These changes will not influence the content and framework of the paper. Special thanks to you for your good comments. We appreciate for Editors/Reviewers’ warm work earnestly, and hope that the correction will meet with approval. Once again, thank you very much for your comments and suggestions.

Hai-min Lei

Aug 16, 2019

Reviewer 2 Report

The submitted manuscript argues for an improved process to replace classic drug development by leading compound discovery strategy based on traditional Chinese medicine (TCM) formulae and pharmacochemistry combination. Authors provide betulinic acid derivative (BA-12) as an example and validate its effect on angiogenesis based on induced metabolic changes.

The authors have an extended original input. The only point to criticize is that the input is really much, where every little part might be questioned.

A new aims oriented production of drug based on the knowledge of plant extracts and further modification might provide drugs that are effective on both anti-angiogenesis and anti-cancer tracks, at the same time provide a limited toxicity on the C. Elegant based assay. The authors provide a hypothesis suggest a material which must be an object of detailed investigation both of them and of other researchers. Except the angiogenesis results which are based on more models and several aspects, all other components could be addressed starting with the cancer cell line - how much is it representative, how much the flow cytometry DNA results different, it is not easy to see the differences, how much apoptosis is the correct measure of the effects, etc. May be the effects could be summarized in a table with micro molar concentrations to compare the effects especial for angiogenesis and toxicity, which are the two highlights in this article.

In all, this is a good work, authors have invested a lot and it is highly significant and relevant to your journal.

Author Response

Dear Editor and Reviewer,

Thank you for your letter and for the reviewers’ comments concerning our manuscript entitled “BA-12 inhibits angiogenesis via glutathione metabolism activation” (ID: ijms-563296). Those comments are all valuable and very helpful for revising and improving our paper, as well as the important guiding significance to our researches. We have studied comments carefully and have made correction which we hope meet with approval. Revised portion are marked using “Track Changes” in the paper. The point to point responds to the reviewer’s comments are listed as follows:

Responds to the reviewer’s comments:  

Reviewer 2

The authors have an extended original input. The only point to criticize is that the input is really much, where every little part might be questioned.

A new aims oriented production of drug based on the knowledge of plant extracts and further modification might provide drugs that are effective on both anti-angiogenesis and anti-cancer tracks, at the same time provide a limited toxicity on the C. Elegant based assay. The authors provide a hypothesis suggest a material which must be an object of detailed investigation both of them and of other researchers. Except the angiogenesis results which are based on more models and several aspects, all other components could be addressed starting with the cancer cell line - how much is it representative, how much the flow cytometry DNA results different, it is not easy to see the differences, how much apoptosis is the correct measure of the effects, etc. May be the effects could be summarized in a table with micro molar concentrations to compare the effects especial for angiogenesis and toxicity, which are the two highlights in this article.

Response:

Thank you very much for your beneficial comments making our manuscript more convincing. We tried to propose one drug discovery strategy based on TCM compatibility theory and combination principle, taking one effective ligustrazine-betulinic acid derivative (BA-12) as an example. Many experiments were carried out to try to express this strategy more objectively. We try our best to correct problems in this manuscript, and thank you for your comments again. It is worth mentioning that one table was added as follows making reference to your comments, which makes the effects of BA-12 more readable. As well, considering the Reviewer’s suggestion, the other details of experiments have been revised using “Track Changes” in the manuscript.

Table 1. Effects of BA-12 on apoptosis and cell cycle of T24 cells.
Group	Apoptosis Ratios %	G0/G1 %	S %	G2/M %
Control	5.7±1.2	62.5±6.7	9.4±1.8	27.6±3.4
Dissolvent	7.2±1.5	62.1±5.1	9.2±1.3	29.0±3.8
Dovitinib (2.5 μM)	25.5±5.1**	50.8±4.2*	11.1±1.5	37.5±4.4*
BA-12 (2.5 μM)	20.3±4.2*	51.7±4.4*	10.3±1.6	38.7±4.0*

ANOVA with the post hoc test was used to calculate the significance of the differences, * and ** represents p < 0.05, and p < 0.01 compared with the dissolvent group. Experiments were executed 3 times. Results are displayed as the mean ± S.D.

We tried our best to improve the manuscript and made some changes in the manuscript. These changes will not influence the content and framework of the paper. Special thanks to you for your good comments. We appreciate for Editors/Reviewers’ warm work earnestly, and hope that the correction will meet with approval. Once again, thank you very much for your comments and suggestions.

Hai-min Lei

Aug 16, 2019