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Investigations into the Fate of Inhaled Salmon Calcitonin at the Respiratory Epithelial Barrier

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ABSTRACT

Purpose

The fate of inhaled salmon calcitonin (sCT) at the respiratory epithelial barrier was studied with particular emphasis on enzymatic degradation by trypsin, chymotrypsin, and neutrophil elastase.

Methods

Degradation of sCT was assessed by HPLC in cell homogenate, supernatant and intact monolayers of human respiratory epithelial cells (hBEpC, Calu-3, 16HBE14o-, A549) and Caco-2 as comparison at 37°C for 2 h. Breakdown of sCT by trypsin, chymotrypsin and neutrophil elastase was investigated. The presence of enzymes in cell supernatant and homogenate was studied by immunoblot and enzyme activity by model substrate assay. Transport studies across Calu-3 monolayers were performed.

Results

sCT concentration remained unchanged over 2 h, when incubated in supernatant or with cell monolayers, independent of cell type studied. When cell homogenates were used, sCT concentrations were reduced to varying extents. sCT was degraded when incubated with enzymes alone. Western blot revealed abundance of all proteinases in cell homogenates and weaker expression in supernatants. Transport studies indicated net-absorptive sCT translocation; presence of bacitracin resulted in increased amount of sCT in receiver compartments.

Conclusions

Epithelial proteases play a role in the disposition of sCT after pulmonary delivery.

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ACKNOWLEDGEMENTS & DISCLOSURES

This work was funded by a Strategic Research Cluster grant (07/SRC/B1154) under the National Development Plan co-funded by EU Structural Funds and Science Foundation Ireland. S.T.B. is funded by an IRCSET Government of Ireland Postgraduate Scholarship in Science, Engineering and Technology. U.B. is funded by the DFG Forschergruppe Nanohale 627.

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Authors and Affiliations

  1. School of Pharmacy and Pharmaceutical Sciences, Trinity College Dublin, Dublin 2, Ireland

    Leonie Baginski, Frederic Tewes, Stephen T. Buckley, Anne Marie Healy & Carsten Ehrhardt

  2. Department of Pharmaceutical Technology and Biopharmaceutics, Philipps University Marburg, Marburg, Germany

    Leonie Baginski & Udo Bakowsky

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  1. Leonie Baginski
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  2. Frederic Tewes
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  3. Stephen T. Buckley
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  4. Anne Marie Healy
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  5. Udo Bakowsky
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  6. Carsten Ehrhardt
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Correspondence to Carsten Ehrhardt.

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Baginski, L., Tewes, F., Buckley, S.T. et al. Investigations into the Fate of Inhaled Salmon Calcitonin at the Respiratory Epithelial Barrier. Pharm Res 29, 332–341 (2012). https://doi.org/10.1007/s11095-011-0553-z

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  • DOI: https://doi.org/10.1007/s11095-011-0553-z

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