Mannitol hemihydrate in lyophilized protein formulations: Impact of its dehydration during storage on sucrose crystallinity and protein stability

Jayesh Sonje, Seema Thakral, Brendan Mayhugh, Gregory Sacha, Steve Nail, Jayasree Srinivasan, Raj Suryanarayanan

Research output: Contribution to journalArticlepeer-review

Abstract

The high propensity of mannitol to crystallize in frozen solutions along with its high eutectic temperature enabling higher primary drying temperatures makes it a good bulking agent. In protein formulations, addition of a sugar (sucrose) that has the ability to remain amorphous throughout processing as well as storage is imperative to retain the protein in its native state. It is well known that in the presence of amorphous excipients and protein, mannitol can crystallize as a mixture of anhydrous polymorphs - α-, β- and δ-forms and a hemihydrate form [mannitol hemihydrate (MHH); C6H14O6·0.5H2O]. The conditions of formation of MHH due to processing and formulation variables are well established in the literature. However, MHH's dehydration kinetics on storage and its impact on the stability of a protein has not been systematically evaluated. The overall objective was to identify conditions (temperature and humidity) at which MHH can dehydrate on storage and the consequences of the release of associated water on sucrose phase behavior and protein stability. In a mannitol-sucrose-protein lyophile, the purpose of this study was (i) to investigate the dehydration behavior of MHH (ii) to determine the influence of dehydration on sucrose crystallization and (iii) the effect of moisture released due to MHH dehydration on model protein (Bovine serum albumin, BSA or Human serum albumin, HSA) aggregation. MHH dehydration and sucrose crystallization was observed in cases where the relative humidity was ≥ 55% (open vials). A relative humidity of ≤ 33% RH prevented MHH dehydration while retaining sucrose amorphous. No protein aggregation was observed irrespective of presence of MHH or its dehydration.

Original languageEnglish (US)
Article number121974
JournalInternational journal of pharmaceutics
Volume624
DOIs
StatePublished - Aug 25 2022

Bibliographical note

Funding Information:
The authors acknowledge funding from the William and Mildred Peters Endowment Fund. We thank Andrew Strongrich, Hallie Harrison and Zack Mora from Lyohub, Purdue University , USA for their generous help in conducting lighthouse FMS experiments. Parts of this work were carried out in the Characterization Facility, University of Minnesota, which receives partial support from the NSF through the MRSEC (award number DMR-2011401) and the NNCI (award number ECCS-2025124) programs. Use of the Advanced Photon Source was supported by U.S. Department of Energy, Office of Science, Office of Basic Energy Science, under Contract No. DE-AC02-06CH11357. We thank Dr. Wenqian Xu and Dr. Andrey Yakovenko for their help during the beamline experiments.

Funding Information:
The authors acknowledge funding from the William and Mildred Peters Endowment Fund. We thank Andrew Strongrich, Hallie Harrison and Zack Mora from Lyohub, Purdue University, USA for their generous help in conducting lighthouse FMS experiments. Parts of this work were carried out in the Characterization Facility, University of Minnesota, which receives partial support from the NSF through the MRSEC (award number DMR-2011401) and the NNCI (award number ECCS-2025124) programs. Use of the Advanced Photon Source was supported by U.S. Department of Energy, Office of Science, Office of Basic Energy Science, under Contract No. DE-AC02-06CH11357. We thank Dr. Wenqian Xu and Dr. Andrey Yakovenko for their help during the beamline experiments.

Publisher Copyright:
© 2022 Elsevier B.V.

Keywords

  • Freeze-drying
  • Mannitol
  • Mannitol hemihydrate dehydration
  • Moisture content
  • Phase transitions
  • Protein aggregation
  • Sucrose

PubMed: MeSH publication types

  • Journal Article

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