Pharmaceutical Research

, Volume 22, Issue 11, pp 1978–1985

Influence of the Active Pharmaceutical Ingredient Concentration on the Physical State of Mannitol—Implications in Freeze-Drying

  • Xiangmin Liao
  • Rajesh Krishnamurthy
  • Raj Suryanarayanan
Research Paper

Purpose

The aim of this study was to investigate the effect of the concentration of the active pharmaceutical ingredient on the physical state of mannitol in frozen aqueous systems.

Methods

A human monoclonal antibody was used as the model protein. Mannitol and sucrose were used as the bulking agent and the lyoprotectant, respectively. The thermal behavior of frozen mannitol–sucrose solutions during and after annealing, in the absence and presence of the protein, were characterized by low-temperature powder X-ray diffractometry and differential scanning calorimetry. The influence of the protein on the crystallization behavior of mannitol was also evaluated.

Results

The excipient concentration had a pronounced effect on the glass transition temperature of maximally freeze-concentrated amorphous phase (Tg′). At fixed excipient compositions, the protein had no effect on the Tg′ if the protein concentration was ≤20 mg/ml. However, at higher protein concentrations, there was a marked increase in Tg′ as a function of protein concentration. The inhibitory effect of the protein on mannitol crystallization was concentration dependent and was directly evident from X-ray diffractometry experiments. Annealing facilitated both mannitol nucleation and crystal growth even in the presence of the protein.

Conclusions

The ratio of mannitol to sucrose and the protein concentration have an impact on the Tg′ and may therefore influence the primary drying temperature. The protein inhibits both the nucleation and growth of mannitol crystals and this effect seems to be concentration dependent. The presence of the protein and the protein concentration dictate the processing conditions, i.e., annealing time, annealing temperature, and primary drying temperature.

Key Words

annealing lyophilization mannitol protein sucrose 

References

  1. 1.
    Rey, L., May, J. C. 2004Freeze-Drying/Lyophilization of pharmaceutical and Biological ProductsMarcel DekkerNew YorkGoogle Scholar
  2. 2.
    Wang, W. 2000Lyophilization and development of solid protein pharmaceuticalsInt. J. Pharm.203160CrossRefPubMedGoogle Scholar
  3. 3.
    Johnson, R. E., Kirchhoff, C. F., Gaud, H. T. 2002Mannitol–sucrose mixtures—versatile formulations for protein lyophilizationJ. Pharm. Sci.91914922CrossRefPubMedGoogle Scholar
  4. 4.
    Kasraian, K., Spitznagel, T. M., Juneau, J. A., Yim, K. 1998Characterization of the sucrose/glycine/water system by differential scanning calorimetry and freeze-drying microscopyPharm. Dev. Technol.3233239PubMedGoogle Scholar
  5. 5.
    Lueckel, B., Helk, B., Bodmer, D., Leuenberger, H. 1998Effects of formulation and process variables on the aggregation of freeze-dried interleukin-6 (IL-6) after lyophilization and on storagePharm. Dev. Technol.3337346PubMedGoogle Scholar
  6. 6.
    Wittaya-Areekul, S., Needham, G. F., Milton, N., Roy, M. L., Nail, S. L. 2002Freeze-drying of tert-butanol/water cosolvent systems: a case report on formation of a friable freeze-dried powder of tobramycin sulfateJ. Pharm. Sci.9111471155CrossRefPubMedGoogle Scholar
  7. 7.
    Kim, A. I., Akers, M. J., Nail, S. L. 1998The physical state of mannitol after freeze-drying: effects of mannitol concentration, freezing rate, and a noncrystallizing cosoluteJ. Pharm. Sci.87931935CrossRefPubMedGoogle Scholar
  8. 8.
    Chang, B. S., Randall, C. S. 1992Use of subambient thermal analysis to optimize protein lyophilizationCryobiology29632 656CrossRefGoogle Scholar
  9. 9.
    Telang, C., Yu, L., Suryanarayanan, R. 2003Effective inhibition of mannitol crystallization in frozen solutions by sodium chloridePharm. Res.20660667CrossRefPubMedGoogle Scholar
  10. 10.
    Torrado, S., Torrado, S. 2002Characterization of physical state of mannitol after freeze-drying: effect of acetylsalicylic acid as a second crystalline cosoluteChem. Pharm. Bull.50567570CrossRefPubMedGoogle Scholar
  11. 11.
    Reilly, R. M., Sandhu, J., Alvarez-Diez, T. M., Gallinger, S., Kirsh, J., Stern, H. 1995Problems of delivery of monoclonal antibodies. Pharmaceutical and pharmacokinetic solutionsClin. Pharmacokinet.28126142PubMedGoogle Scholar
  12. 12.
    Izutsu, K., Yoshioka, S., Terao, T. 1993Decreased protein-stabilizing effects of cryoprotectants due to crystallizationPharm. Res.1012321237CrossRefPubMedGoogle Scholar
  13. 13.
    Izutsu, K., Yoshioka, S., Terao, T. 1994Effect of mannitol crystallinity on the stabilization of enzymes during freeze-dryingChem. Pharm. Bull.4258PubMedGoogle Scholar
  14. 14.
    Kreilgaard, L., Frokjaer, S., Flink, J. M., Randolph, T. W., Carpenter, J. F. 1998Effects of additives on the stability of recombinant human factor XIII during freeze-drying and storage in the dried solidArch. Biochem. Biophys.360121134CrossRefPubMedGoogle Scholar
  15. 15.
    Lam, X. M., Duenas, E. T., Cleland, J. L. 2001Encapsulation and stabilization of nerve growth factor into poly(lactic-co-glycolic) acid microspheresJ. Pharm. Sci.9013561365CrossRefPubMedGoogle Scholar
  16. 16.
    Cavatur, R. K., Vemuri, N. M., Pyne, A., Chrzan, Z., Toledo-Velasquez, D., Suryanarayanan, R. 2002Crystallization behavior of mannitol in frozen aqueous solutionsPharm. Res.19894900CrossRefPubMedGoogle Scholar
  17. 17.
    Passot, S., Fonseca, F., Alarcon-Lorca, M., Rolland, D., Marin, M. 2005Physical characterization of formulations for the development of two stable freeze-dried proteins during driedand liquid storageEur. J. Pharm. Biopharm.60335348CrossRefPubMedGoogle Scholar
  18. 18.
    Chang, L., Tang, X., Pikal, M. J., Milton, N., Thomas, L. 1999The origin of multiple glass transitions in frozen aqueous solutionsProc. NATAS Annu. Conf. Therm. Anal. Appl.27624628Google Scholar
  19. 19.
    Ablett, S., Izzard, M. J., Lillford, P. J. 1992Differential scanning calorimetric study of frozen sucrose and glycerol solutionsJ. Chem. Soc. Faraday Trans.88789794CrossRefGoogle Scholar
  20. 20.
    Izzard, M. J., Ablett, S., Lillford, P. J., Hill, V. L., Groves, I. F. 1996A modulated differential scanning calorimetric study. Glass transitions occurring in sucrose solutionsJ. Therm. Anal.4714071418CrossRefGoogle Scholar
  21. 21.
    Tang, X., Pikal, M. J. 2004Design of freeze-drying processes for pharmaceuticals: practical advicePharm. Res.21191200CrossRefPubMedGoogle Scholar
  22. 22.
    Martini, A., Kume, S., Crivellente, M., Artico, R. 1997Use of subambient differential scanning calorimetry to monitor the frozen-state behavior of blends of excipients for freeze-dryingPDA J. Pharm. Sci. Technol.516267PubMedGoogle Scholar
  23. 23.
    Her, L. M., Nail, S. L. 1994Measurement of glass transition temperatures of freeze-concentrated solutes by differential scanning calorimetryPharm. Res.115459CrossRefPubMedGoogle Scholar
  24. 24.
    Shalaev, E. Y., Franks, F. 1995Structural glass transitions and thermophysical processes in amorphous carbohydrates and their super-saturated solutionsJ. Chem. Soc. Faraday Trans.9115111517CrossRefGoogle Scholar
  25. 25.
    Lueckel, B., Bodmer, D., Helk, B., Leuenberger, H. 1998Formulations of sugars with amino acids or mannitol—influence of concentration ratio on the properties of the freeze-concentrate and the lyophilizatePharm. Dev. Technol.3325336PubMedGoogle Scholar
  26. 26.
    Chatterjee, K., Shalaev, E. Y., Suryanarayanan, R. 2005Partially crystalline systems in lyophilization: II. Withstanding collapse at high primary drying temperatures and impact on protein activity recoveryJ. Pharm. Sci.94809820CrossRefPubMedGoogle Scholar
  27. 27.
    Searles, J. A., Carpenter, J. F., Randolph, T. W. 2001Annealing to optimize the primary drying rate, reduce freezing-induced drying rate heterogeneity, and determine Tg′ in pharmaceutical lyophilizationJ. Pharm. Sci.90872887CrossRefPubMedGoogle Scholar
  28. 28.
    Pyne, A., Surana, R., Suryanarayanan, R. 2002Crystallization of mannitol below Tg′ during freeze-drying in binary and ternary aqueous systemsPharm. Res.19901908CrossRefPubMedGoogle Scholar
  29. 29.
    Yu, L., Milton, N., Groleau, E. G., Mishra, D. S., Vansickle, R. E. 1999Existence of a mannitol hydrate during freeze-drying and practical implicationsJ. Pharm. Sci.88196198CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • Xiangmin Liao
    • 1
  • Rajesh Krishnamurthy
    • 2
  • Raj Suryanarayanan
    • 1
  1. 1.Department of PharmaceuticsUniversity of MinnesotaMinneapolisUSA
  2. 2.Human Genome Science, Inc.RockvilleUSA

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