Characterisation of protein stability in rod-insert vaginal rings

Int J Pharm. 2012 Jul 1;430(1-2):89-97. doi: 10.1016/j.ijpharm.2012.03.036. Epub 2012 Apr 2.

Abstract

A major goal in vaccine development is elimination of the 'cold chain', the transport and storage system for maintenance and distribution of the vaccine product. This is particularly pertinent to liquid formulation of vaccines. We have previously described the rod-insert vaginal ring (RiR) device, comprising an elastomeric body into which are inserted lyophilised, rod-shaped, solid drug dosage forms, and having potential for sustained mucosal delivery of biomacromolecules, such as HIV envelope protein-based vaccine candidates. Given the solid, lyophilised nature of these insert dosage forms, we hypothesised that antigen stability may be significantly increased compared with more conventional solubilised vaginal gel format. In this study, we prepared and tested vaginal ring devices fitted with lyophilised rod inserts containing the model antigen bovine serum albumin (BSA). Both the RiRs and the gels that were freeze-dried to prepare the inserts were evaluated for BSA stability using PAGE, turbidimetry, microbial load, MALDI-TOF and qualitative precipitate solubility measurements. When stored at 4 °C, but not when stored at 40 °C/75% RH, the RiR formulation offered protection against structural and conformational changes to BSA. The insert also retained matrix integrity and release characteristics. The results demonstrate that lypophilised gels can provide relative protection against degradation at lower temperatures compared to semi-solid gels. The major mechanism of degradation at 40 °C/75% RH was shown to be protein aggregation. Finally, in a preliminary study, we found that addition of trehalose to the formulation significantly reduces the rate of BSA degradation compared to the original formulation when stored at 40 °C/75% RH. Establishing the mechanism of degradation, and finding that degradation is decelerated in the presence of trehalose, will help inform further development of RiRs specifically and polymer based freeze-dried systems in general.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Intravaginal
  • Chemistry, Pharmaceutical
  • Chromatography, High Pressure Liquid
  • Circular Dichroism
  • Delayed-Action Preparations
  • Drug Carriers
  • Drug Compounding
  • Drug Delivery Systems / instrumentation*
  • Drug Stability
  • Elastomers / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Equipment Contamination
  • Equipment Design
  • Excipients / chemistry
  • Female
  • Freeze Drying
  • Gels
  • Humans
  • Humidity
  • Hypromellose Derivatives
  • Methylcellulose / analogs & derivatives
  • Methylcellulose / chemistry
  • Nephelometry and Turbidimetry
  • Polysorbates / chemistry
  • Protein Conformation
  • Protein Stability
  • Serum Albumin, Bovine / administration & dosage
  • Serum Albumin, Bovine / chemistry*
  • Solubility
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Technology, Pharmaceutical / methods
  • Temperature
  • Thermogravimetry
  • Time Factors
  • Trehalose / chemistry
  • Vaccines / administration & dosage
  • Vaccines / chemistry*

Substances

  • Delayed-Action Preparations
  • Drug Carriers
  • Elastomers
  • Excipients
  • Gels
  • Polysorbates
  • Vaccines
  • Serum Albumin, Bovine
  • Hypromellose Derivatives
  • Methylcellulose
  • Trehalose