Targeting TGF-ß in the Central Nervous System: Assessment of Cynomolgus Monkey—Toxicity and Pharmacokinetics for an LNA-Antisense Oligonucleotide
, Siw Johannesen 1,†, Tim-Henrik Bruun 1,‡ and Ulrich Bogdahn 1,*,‡Round 1
Reviewer 1 Report
Sebastian Peters et al assessed the toxicity and pharmacokinetics of LNA-ASO in Cynomolgus monkeys.
Below are the comments and provide the information in the manuscript:
- There are several repeated sections/paragraphs present in the manuscript. This manuscript needs thorough revision to minimize the repeated sections/paragraphs. Eg. Histology, special processing, NVP-13 concentration measurement, plasma, CSF.
- The authors need to correct the manuscript for spelling mistakes and grammar.
- The source of Cynomolgus monkeys should be mentioned in the Animals section of Materials and methods.
- References need to be cited for Lines 128-133.
- The authors should include the blood volume collection for different phase studies like toxicity and pharmacokinetics.
- HPLC chromatograms for bioanalytical samples need to be provided in the manuscript. The authors should include the bioanalytical method and sample processing procedure for NVP13. There are no proper method details mentioned like mobile phase, gradient/isocratic elution, retention time, run time, etc.
- There are several repeated sections/paragraphs present in the manuscript. This manuscript needs thorough revision to minimize the repeated sections/paragraphs.
- Figures and Tables legends should be modified with detailed information. There are several mistakes in the tables and figures legends.
- Table 2 and Figure 4 are the same and Table 2 should be removed. Figures 7 and 8 are the same and Figure 7 should be deleted.
- Is NVP13 shows dose linearity in terms of pharmacokinetic parameters like Cmax, AUC for plasma, CSF, and tissue samples?
- Table 1 is not properly represented. Page 10 tables should be combined in one table for Table 1.
- Several studies for different parameters were performed in this manuscript, but the authors didn’t show the data for several parameters. The authors should include some of the data in the manuscript like histology.
- The authors didn’t show any data related to 8 mg/dose of ASO. How do the authors select 8 mg/dose for subsequent GLP studies? What would be the reason for the selection of this dose?
Author Response
Please see the attachment
Author Response File: 
Author Response.pdf
Reviewer 2 Report
The study of Peters et al describes a safety study of a new Antisense oligonucleotide (ASO) used to downregulate TNF-alpha. Additionally, a new and very interesting approach is tested to evaluate the toxicity and pharmacokinetics of the new LNA-ASO in Cynomolgus Monkey. This new approach is conducted under non-GLP conditions and comprises 3 phases conducted in only 26 animals. The advantages of this approach are obvious, it is more time and cost-effective and a lower number of animals are needed. It might be suitable to evaluate after intrathecal administrations the maximum tolerated dose, the pharmacokinetics and to determine the dose range for a 13-week GLP repeated intrathecal dose toxicity study. The study is straightforward and well written, it has a clear experimental design and is in my option in the clear scope of the journal.
I have some suggestions/issues which should be addressed before this paper is suitable for publication:
Unfortunately, and this is my main criticism, the authors want to investigate two new parameters: 1) the toxicity and the pharmacokinetics of a new LNA-ASO and 2) a new method to evaluate the toxicity and the pharmacokinetics.
In principle toxicity and pharmacokinetics can only be proven by an established method or by a method that is evaluated. For evaluation, it is necessary to compare obtained results by a new method with a formerly established one. By mixing these two things no exact toxicity nor a method evaluation can be done.
This is in particular critical as low animal numbers are used. E.g. For the first phase an n of 1 per gender. Were similar results obtained by a GLP used method?
There is no doubt that this new method follows a clear rationale and the experiments appear scientifically sound. However, the comparison and therefore the proof is missing. The authors refer to a manuscript that is “in preparation” (line 92/93) where GLP-based methods are used. Unfortunately, without these experiments being visible no final recommendation can be given.
In my opinion, this paper has a nice impact in the field, it is interesting for readers of the journal but without the above-mentioned data, a publication is not possible. I therefore highly encourage the authors to fuse these two manuscripts or to publish them back to back in this journal if the size of the manuscript exceeds the journal guidelines. It is necessary to compare the data of these 2 manuscripts and have them accessible to the reader. Because of this principle concern, I cannot recommend the publication of the MS without this lacking data. As the manuscript is of great interest I suggest exceeding the deadline for a revision that the authors are able for e.g. publishing these two manuscripts back to back.
Author Response
Please see the attachment
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
The authors significantly improved the manuscript and provided point-to-point responses for all comments.
- Figure 6 legend is not matching with the data provided in the images. Figures 6 and 7 have the same information and figure 6 should be deleted.
Author Response
Point 1: Figure 6 legend is not matching with the data provided in the images. Figures 6 and 7 have the same information and figure 6 should be deleted.
Response 1: Figure 6 was replaced with the correct Figure content. See new manuscript upload.
Reviewer 2 Report
As I mentioned in my previous report, without the GLP data, which have to be compared with the novel results, the new method and its suitability cannot be judged. I, therefore, recommended publishing these data “back to back”, which was done by the authors. See response below.
“ Response 1: The publication of the GLP data "back to back", as requested by the reviewer, had already been intended and is now initiated and taking place within MDPI in the journal "pharmaceutics”. “
However, without attaching this paper or without adding data from this manuscript I am still in the situation like before. Without this information, I cannot recommend publishing the paper.
Author Response
Point 1: However, without attaching this paper or without adding data from this manuscript I am still in the situation like before. Without this information, I cannot recommend publishing the paper.
Response 1: As suggested by the reviewer, we have attached the current version of the pharmaceutics manuscript under review (pharmaceutics-1480672).
Author Response File: Author Response.pdf
Round 3
Reviewer 2 Report
In combination with the data submitted to Pharmaceutics (MDPI), which were now attached in this review process, I can recommend publishing the manuscript in its present form.
