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Effect of Sugars on the Molecular Motion of Freeze-Dried Protein Formulations Reflected by NMR Relaxation Times

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ABSTRACT

Purpose

To relate NMR relaxation times to instability-related molecular motions of freeze-dried protein formulations and to examine the effect of sugars on these motions.

Methods

Rotating-frame spin-lattice relaxation time (T1ρ) was determined for both protein and sugar carbons in freeze-dried lysozyme-sugar (trehalose, sucrose and isomaltose) formulations using solid-state 13C NMR.

Results

The temperature dependence of T1ρ for the lysozyme carbonyl carbons in lysozyme with and without sugars was describable with a model that includes two different types of molecular motion with different correlation times (τc) for the carbon with each τc showing Arrhenius temperature dependence. Both relaxation modes have much smaller relaxation time constant (τc) and temperature coefficient (Ea) than structural relaxation and may be classified as β-relaxation and γ-relaxation. The τc and Ea for γ-relaxation were not affected by sugars, but those for β-relaxation were increased by sucrose, changed little by trehalose, and decreased by isomaltose, suggesting that the β-mobility of the lysozyme carbonyl carbons is decreased by sucrose and increased by isomaltose.

Conclusion

T1ρ determined for the lysozyme carbonyl carbons can reflect the effect of sugars on molecular mobility in lysozyme. However, interpretation of relaxation time data is complex and may demand data over an extended temperature range.

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Notes

  1. This statement is equivalent to stating that the motion represented by the relaxation process may have nothing directly to do with the motion required for degradation, if the “coupling coefficient” relating a relaxation time constant for a given microstate with degradation within that microstate, which has been discussed in a reference (6), is much less than unity.

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Authors and Affiliations

  1. School of Pharmacy, University of Connecticut, Storrs, Connecticut, 06269–3092, USA

    Sumie Yoshioka, Kelly M. Forney & Michael J. Pikal

  2. National Institute of Health Sciences, Setagaya, Tokyo, 158–8501, Japan

    Yukio Aso

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  1. Sumie Yoshioka
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  2. Kelly M. Forney
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  3. Yukio Aso
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  4. Michael J. Pikal
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Correspondence to Sumie Yoshioka.

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Yoshioka, S., Forney, K.M., Aso, Y. et al. Effect of Sugars on the Molecular Motion of Freeze-Dried Protein Formulations Reflected by NMR Relaxation Times. Pharm Res 28, 3237–3247 (2011). https://doi.org/10.1007/s11095-011-0512-8

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  • DOI: https://doi.org/10.1007/s11095-011-0512-8

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