Abstract Details

Title A Cell-Penetrating Peptide (CPP) Did Not Decrease 150-kDa BoNT/A Toxin Adsorption to Surfaces or Increase Toxin Potency or Duration in a Prototype Formulation

Topic General Neurology

Presentation(s) General Neurology Posters (7:00 AM-5:00 PM)

Poster/Presentation
Number 088

Objective To evaluate the effect of a cell-penetrating peptide (CPP) in a prototype botulinum neurotoxin type A (BoNT/A) formulation using toxin adsorption, in vivo potency, and duration analyses.

Background Several BoNT/A drugs are approved or under development for numerous indications. Unit potencies of these drugs are not interchangeable due to various factors, including unique manufacturing processes and drug formulations. Various excipients, in different combinations, have been used to stabilize the drug in the vial and prevent adsorption to surfaces.

Design/Methods A CPP with the sequence RKKRRQRRRG-[K]15-GRKKRRQRRR was synthesized. Size exclusion chromatography compared the adsorption of 150-kDa BoNT/A toxin formulated in either potassium phosphate/NaCl buffer (pH 6.0 and 7.2) containing polysorbate 20, 23.5 μg/mL CPP with and without polysorbate 20, human serum albumin (HSA), or in buffer alone. Mouse digit abduction score (DAS) testing compared the potency and duration of 150-kDa BoNT/A toxin at an approximate ED₅₀ dose formulated in histidine buffer/trehalose buffer (pH 6.0) containing polysorbate 20, 0.235 μg CPP/unit BoNT/A with or without polysorbate 20, or in bovine serum albumin (BSA)/0.9% NaCl.

Results Toxin adsorption to the glass vial surface was observed in buffer control and CPP-only solutions at 7 hours, the first assessed time point, both with toxin recovery of <64%. At 14 and 21 hours, these buffer control and CPP-alone samples had decreased toxin recoveries of <42%; samples containing HSA or polysorbate 20 continued to display toxin recoveries of >95%. In the mouse DAS assay, BoNT/A prepared in polysorbate formulations with or without CPP exhibited similar, predictive DAS efficacy (>ED₅₀) and duration versus the formulation in BSA/0.9% NaCl. In contrast, the formulation in CPP alone, without polysorbate 20, demonstrated decreased potency (<<ED₂₅) and duration.

Conclusions Inclusion of a cell-penetrating peptide in a BoNT/A formulation neither prevents toxin adsorption nor increases toxin potency or duration in the mouse DAS assay.

Authors/Disclosures
PRESENTER	No disclosure on file
	No disclosure on file
Ron S. Broide, PhD (Allergan Aesthetics / AbbVie)	Dr. Broide has received personal compensation for serving as an employee of Allergan Aesthetics / AbbVie.
	No disclosure on file
	No disclosure on file
	No disclosure on file
	No disclosure on file
	No disclosure on file
	No disclosure on file
Mariana Nelson, PhD (AbbVie/Allergan US Medical Affairs)	Dr. Nelson has received personal compensation for serving as an employee of AbbVie. Dr. Nelson has stock in AbbVie Inc.
Lance E. Steward, PhD	Dr. Steward has received personal compensation for serving as an employee of Allergan Aesthetics an AbbVie Company. Dr. Steward has received stock or an ownership interest from Allergan Aesthetics an AbbVie Company.
	No disclosure on file
Mitchell F. Brin, MD, FAAN (Abbvie / UC Irvine)	Dr. Brin has received personal compensation for serving as an employee of Allergan. Dr. Brin has stock in Allergan.
	No disclosure on file

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