Abstract Committee Review

In the introduction:

please reconsider whether Xeomin should be described as a new product.

In, the same sentence, I suggest to rephrase the second half for clarity to: ...are commercialized with claims of various duration-of-effect, potency, or spread.  However, dose unit equivalency has not been established.

Before Relabotulinumtoxin, change comma to period.

Methods:

Since a subjective assay was used to determine bio-activity, please include information on whether the injector and observer were blinded.

For the volumetrical determination of spread in porcine muscle, please describe what exactly was measured.

Results:

For relative bioactivity, please include SD or SEM if available, and specify statistical significance

Please clarify the meaning of equivalent bioanalytically determined dose for the assay that injected 20U/kg of INCO vs DAXI (i.e. were label units used or were units based on the DAS assay? What mass of each was injected?)

Please confirm the statement that INCO lasted in a 10-day longer duration in mice. The total paralysis after IM injection in mice is 2-3 weeks only, so 10 days seems like a huge difference.

thank you for the committee’s comments and suggestions to abstract 0129 which we have incorporated in the revised version attached. Changes in the abstract are in track changes mode and the line numbers provided underneath in this email refer to the track changes version. As we agree with all comments the responses in red here are fyi only.

Introduction:

• please reconsider whether Xeomin should be described as a new product. We have deleted like Xeomin from the sentence on new products in Intro line 4.
• In, the same sentence, I suggest to rephrase the second half for clarity to: ...are commercialized with claims of various duration-of-effect, potency, or spread.  However, dose unit equivalency has not been established. Revised as suggested in Intro line 4-6.
• Before Relabotulinumtoxin, change comma to period. Revised as suggested in Intro line 8.

Methods:

• Since a subjective assay was used to determine bio-activity, please include information on whether the injector and observer were blinded. We have added unblinded to the DAS in Methods line 5. It is correct that the DAS is a subjective assay, but the contract lab which did the injections and evaluations is not interested in the sample identities that they administer. It is, thus, for us more important to make sure the same person evaluates the degree of paralysis over the entire study period.
• For the volumetrical determination of spread in porcine muscle, please describe what exactly was measured. We have added the stain bromophenole blue in Methods line 8, the blue stained tissue area was measured in each cryo section and the volume calculated from the cryo section stack, line 9-10. The technique will be illustrated on the poster.

Results:

• For relative bioactivity, please include SD or SEM if available, and specify statistical significance. We have added SD for the relative bioactivity in Results line 3-4.
• Please clarify the meaning of equivalent bioanalytically determined dose for the assay that injected 20U/kg of INCO vs DAXI (i.e. were label units used or were units based on the DAS assay? What mass of each was injected?). Dosing was based on the labeled units for all drug products. The term equivalent bioanalytically determined was misleading and deleted. Mass could be calculated from the NT content provided in Results line 2 and the number of units injected in line 7; the full data will also be in a table on the poster.
• Please confirm the statement that INCO lasted in a 10-day longer duration in mice. The total paralysis after IM injection in mice is 2-3 weeks only, so 10 days seems like a huge difference. Correct, but the duration of effect in mice is dose-dependent while the maximum DAS is limited by the highest possible DAS score of 4, which appears like a classical ceiling effect in the diagram. In our DAS setup 20U/kg of most BoNTAs, corresponding to 0.4U/mouse depending on the body weight, result in a readable DAS for 3-4 weeks, 40U/kg one week longer. We confirm that the duration of paralysis after INCO injection was 31 days vs. 21 days after DAXI injection which has been added in Results line 11.

Please let me know if there are further concerns.

Best regards

Klaus