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Leucine Protects Dry Powders for Inhalation Against Irreversible Moisture-Induced Aggregation.

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Leucine addition to inulin powders prevents moisture-induced aggregation during storage, maintaining physical stability. However, it does not protect proteins reliant on vitrification, suggesting specific applications for biologics delivery.

Keywords:
LDHinulinleucinemoisturespray dryingstorage stabilityvitrificationwater replacementβ-gal

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Area of Science:

  • Pharmaceutical Sciences
  • Materials Science
  • Biotechnology

Background:

  • Pulmonary drug delivery is a key route for respiratory disease treatments using biologics.
  • Spray drying is common for creating inhalable powders, often using amorphous sugars for stabilization.
  • Hydrophobic amino acids like leucine are added to enhance powder dispersibility.

Purpose of the Study:

  • To evaluate leucine's impact on moisture-induced dry powder aggregation and protein stability during storage.
  • To assess the long-term physical and chemical stability of inulin-based powders with and without leucine under varying humidity.

Main Methods:

  • Spray drying of inulin powders with and without 4 wt-% leucine.
  • Storage of powders at various relative humidity (RH) levels (43%, 58%, 69%, 75%) at 20 ± 2 °C for up to 20 days.
  • Assessment of particle aggregation, glass transition temperature (Tg), and protein stability (lactate dehydrogenase and β-galactosidase).

Main Results:

  • Inulin-only powders aggregated irreversibly at RH ≥ 58%, while leucine-containing powders remained intact across all conditions.
  • Protein stability varied: both model proteins retained activity at 43% RH. At 75% RH, β-galactosidase remained stable, but lactate dehydrogenase lost activity.
  • Leucine addition prevented powder aggregation but did not protect proteins dependent on vitrification for stability.

Conclusions:

  • Leucine significantly enhances the physical stability of inulin powders against moisture-induced aggregation.
  • The protective effect of leucine is specific and does not extend to all protein types, particularly those relying on vitrification.
  • Inulin and leucine formulations show potential for reducing stringent storage condition requirements for certain biologics.