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Chemical Triphosphorylation of Oligonucleotides
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Development of stable liquid formulations for oligonucleotides.

Johanna Poecheim1, Kirsten A Graeser2, Joerg Hoernschemeyer2

  • 1Roche Pharmaceutical Development & Supplies, PTD Biologics Europe, F. Hoffmann-La Roche Ltd, Basel, Switzerland; Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland.

European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Fur Pharmazeutische Verfahrenstechnik E.V
|May 27, 2018
PubMed
Summary

This study optimized liquid formulations for single-stranded oligonucleotides (SSOs) with locked nucleic acid modifications (LNA SSOs). Optimal stability was achieved under slightly alkaline conditions, with specific excipients reducing viscosity and avoiding harsh sterilization methods.

Keywords:
Biopharmaceuticals characterizationForced conditionsFormulationOligonucleotidesStability

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Area of Science:

  • Biotechnology and Pharmaceutical Sciences
  • Nucleic Acid Chemistry
  • Drug Delivery Systems

Background:

  • Oligonucleotide therapeutics represent a promising new class of drugs.
  • Developing stable parenteral formulations is crucial for their clinical application.
  • Understanding formulation effects on modified oligonucleotides is essential for drug development.

Purpose of the Study:

  • To investigate the stabilizing and de-stabilizing effects of formulation parameters on LNA SSOs.
  • To identify optimal conditions for liquid formulations of unconjugated and GalNAc-conjugated LNA SSOs.
  • To determine feasible concentrations and terminal sterilization methods for LNA SSO formulations.

Main Methods:

  • Evaluation of various buffer systems, pH levels, and excipients.
  • Assessment of LNA SSO stability under different temperature, mechanical, and oxidative stress conditions.
  • Determination of maximum feasible concentrations for handling and processing.
  • Evaluation of terminal sterilization options.

Main Results:

  • LNA SSOs exhibited maximum stability under slightly alkaline conditions (pH ~8).
  • Spermine and lysine effectively reduced formulation viscosity.
  • Heat treatment and gamma irradiation led to significant LNA SSO degradation.
  • Optimal formulation parameters were identified for enhanced LNA SSO stability.

Conclusions:

  • Formulation optimization is critical for the stability of LNA SSOs.
  • Slightly alkaline conditions and specific excipients are key for stable liquid formulations.
  • Avoiding heat and gamma irradiation is necessary to prevent degradation during terminal sterilization.