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Development of a Flow-Through USP-4 Apparatus Drug Release Assay to Evaluate Doxorubicin Liposomes

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Abstract

Doxil® is a complex parenteral doxorubicin (DOX) liposome formulation approved by the FDA. For generic doxorubicin liposomes, analyzing the release profile of DOX is important for quality control and comparability studies. However, there is no robust standard drug release assay available for doxorubicin liposomes. In this study, we describe a USP-4 apparatus assay capable of discriminating DOX liposomal formulations based on release profile. Establishment of the assay was hindered by limited DOX release from liposomes in physiological conditions at 37°C. The addition of NH4HCO3 to the release media facilitated DOX release proportionally to the salt concentration added but caused precipitation of released drug in USP-4 apparatus. Precipitation of DOX was avoided by adding hydroxypropyl-cyclodextrin (HP-CD) to the release medium. We optimized conditions for DOX release by varying a number of parameters such as: concentration of HP-CD, testing temperature, and concentration of tested samples. The optimized release medium contained: 100 mM NH4HCO3, 75 mM 2-(N-morpholino) ethanesulfonic acid (MES) and 5% w/v HP-CD, 5% w/v sucrose, 0.02% w/v NaN3 (pH 6). The drug release assay was performed at 45°C. The optimized release assay can discriminate between DOX liposomal formulations of different compositions, physicochemical properties, and prepared by different manufacturing methods. This indicates that the assay could be used to compare DOX release from generic DOX formulations to the innovator product Doxil®.

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Acknowledgments

This project was supported by FDA grant U01 FD004893. Yue Yuan was supported by grant 81202481 of National Natural Science for Youth Foundation of China and grant GGJJ2014102 of Scientific Research Foundation for the Returned Overseas Chinese Scholars by Shenyang Pharmaceutical University.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, NCRC, 2800 Plymouth Road, Ann Arbor, Michigan, 48105, USA

    Wenmin Yuan, Rui Kuai, Yue Yuan & Anna Schwendeman

  2. Biointerfaces Institute, University of Michigan, NCRC, 2800 Plymouth Road, Ann Arbor, Michigan, 48105, USA

    Wenmin Yuan, Rui Kuai, Yue Yuan & Anna Schwendeman

  3. ZoneOne Pharma Inc., UCSF Mission Bay, QB3 Garage, Byers Hall, 1700 4th St., Suite 214, MC2522, San Francisco, California, 94158, USA

    Zhipeng Dai, Charles Noble, Mark Hayes & Francis C. Szoka

  4. School of Pharmacy, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang, 110016, People’s Republic of China

    Yue Yuan

  5. Office of Generic Drugs, Food and Drug Administration, 10903 New Hampshire Avenue, Silver Spring, Maryland, 20993, USA

    Nan Zheng & Wenlei Jiang

  6. Department of Bioengineering and Therapeutic Sciences, UCSF, 513 Parnassus Ave., Box 0912, San Francisco, California, 94143, USA

    Francis C. Szoka

Authors
  1. Wenmin Yuan
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  2. Rui Kuai
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  3. Zhipeng Dai
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  4. Yue Yuan
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  5. Nan Zheng
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  6. Wenlei Jiang
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  7. Charles Noble
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  8. Mark Hayes
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  9. Francis C. Szoka
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  10. Anna Schwendeman
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Corresponding author

Correspondence to Anna Schwendeman.

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Disclaimer: The views expressed in this article are those of the authors (Nan Zheng and Wenlei Jiang) and not necessarily those of the Food and Drug Administration (FDA).

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Yuan, W., Kuai, R., Dai, Z. et al. Development of a Flow-Through USP-4 Apparatus Drug Release Assay to Evaluate Doxorubicin Liposomes. AAPS J 19, 150–160 (2017). https://doi.org/10.1208/s12248-016-9958-2

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  • DOI: https://doi.org/10.1208/s12248-016-9958-2

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