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Influence of the Freeze-Drying Process on the Physicochemical and Biological Properties of Pre-heated Amphotericin B Micellar Systems

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Abstract

The moderate heat treatment of amphotericin B (AmB) in its micellar form (M-AmB) results in superaggregates (H-AmB) that present a substantially lower toxicity and similar activity. The aim of this work was to evaluate the H-AmB behavior after a freeze-drying process. H-AmB and M-AmB micelles were evaluated before and after freeze-drying concerning their physicochemical and biological properties by spectrophotometry and activity/toxicity assay, respectively. Four concentrations of M-AmB and H-AmB were studied aiming to correlate their aggregation state and the respective biological behavior: 50 mg L−1, 5 mg L−1, 0.5 mg L−1, and 0.05 mg L−1. Then, potassium leakage and hemoglobin leakage from red blood cells were used to evaluate the acute and chronic toxicity, respectively. The efficacy of M-AmB and H-AmB formulations was assessed by potassium leakage from Candida albicans and by the broth microdilution method. After heating, in addition to an evident turbidity, a slight blueshift from 327 to 323 nm was also observed at the concentrations of 50 and 5 mg L−1 for H-AmB. Additionally, an increase in the absorbance at 323 nm at the concentration of 0.5 mg L−1 was detected. Concerning the toxicity, H-AmB caused significantly lower hemoglobin leakage than M-AmB. These results were observed for H-AmB before and after freeze-drying. However, there was no difference between H-AmB and M-AmB concerning their activity. Accordingly, the freeze-drying cycle did not show any influence on the behavior of heated formulations, highlighting the suitability of such a method to produce a new AmB product with a long shelf life and with both greater efficiency and less toxicity.

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Acknowledgments

The authors wish to thank CAPES for the financial support and Cristália for the generous gift of Fungizone® samples. The authors are also grateful to Glenn Hawes, M.Ed. English from the University of Georgia, for editing this manuscript.

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Authors and Affiliations

  1. Programa de Pós-graduação em Ciências da Saúde, Centro de Ciências da Saúde (CCS), Laboratório de Sistemas Dispersos (LASID), Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN, Brazil

    Scheyla D. V. S. Siqueira, Miguel A. Silva-Filho, Ivonete B. Araújo, Acarilia E. Silva & E. Sócrates T. Egito

  2. Departamento de Farmácia, UFRN, CCS, LASID, Natal, RN, Brazil

    Christian A. Silva, Ivonete B. Araújo & E. Sócrates T. Egito

  3. Departamento de Farmácia, Laboratório de Tecnologia & Biotecnologia Farmacêutica (TecBioFar), UFRN, CCS, Natal, Brazil

    Matheus F. Fernandes-Pedrosa

  4. Departamento de Fármacos e Medicamentos, Faculdade de Ciências Farmacêuticas (UNESP), Rodovia Araraquara, Jaú Km 01, 14801-902, Araraquara, SP, Brazil

    Anselmo G. Oliveira

  5. Laboratório de Sistemas Dispersos, Rua Praia de Areia Branca, 8948, Natal, RN, Brazil, Zip Code: 59094-450

    E. Sócrates T. Egito

Authors
  1. Scheyla D. V. S. Siqueira
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  2. Miguel A. Silva-Filho
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  3. Christian A. Silva
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  4. Ivonete B. Araújo
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  5. Acarilia E. Silva
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  7. Anselmo G. Oliveira
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  8. E. Sócrates T. Egito
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Correspondence to E. Sócrates T. Egito.

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Siqueira, S.D.V.S., Silva-Filho, M.A., Silva, C.A. et al. Influence of the Freeze-Drying Process on the Physicochemical and Biological Properties of Pre-heated Amphotericin B Micellar Systems. AAPS PharmSciTech 15, 612–619 (2014). https://doi.org/10.1208/s12249-014-0085-z

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  • DOI: https://doi.org/10.1208/s12249-014-0085-z

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