Pulmonary and Regional Deposition of Nebulized and Dry Powder Aerosols in Ferrets


The utilization of ferrets as a non-clinical model for disease is rapidly increasing within drug development. Many of these models include respiratory diseases that involve targeted drug delivery via nose-only inhalation. While the deposition patterns within other non-clinical models (mice, rats, canines, and non-human primates) have been well studied, the local and regional deposition of aerosols in ferrets has not been well characterized. Therefore, inhalation aerosols were developed, radiolabeled and the radiolabeling methods validated to support SPECT-CT imaging and quantification of regional deposition within ferrets. The studies were conducted with one liquid formulation and one dry powder formulation (two concentrations of dry powder). Additionally, both aerosols were polydisperse and therefore reflect the majority of pharmaceutical aerosols. Overall, the studies showed lung deposition fractions between 5 and 10% with median aerodynamic particle sizes of 2.5 and 2.8 μm. The lung deposition fraction of the liquid aerosol was ~ 9%, nearly double observed in rats with a similarly sized aerosol. Analysis of respiratory tract (oropharynx, laryngopharynx, trachea, bifurcation area, and lung) deposition indicates increased deposition of the liquid aerosol compared to the dry powder aerosol, however, when this analysis was refined to the pulmonary region (trachea, bifurcation, and lung) the deposition was similar between formulations. These data provide the first description of the regional deposition of inhalation aerosols in ferrets with standard nose-only inhalation procedures. These data can be used for calculations of both total and regional doses within ferret inhalation drug delivery.

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  1. 1.

    Kuehl PJ, Anderson T, Candelaria G, Gershman B, Harlin K, Hesterman JY, et al. Regional particle size dependent deposition of inhaled aerosols in rats and mice. Inhal Toxicol. 2012;24(1):27–35.

    CAS  Article  Google Scholar 

  2. 2.

    Garcia, A., Mack P, Williams S, Fromen C, Shen T, Pillai J, Kuehl PJ, Napier M, Desimon JM, Maynor BW. Microfabricated engineered particle systems for respiratory drug delivery and other pharmaceutical applications. J Drug Deliv 2012;Article ID 941243.

  3. 3.

    Cheng YS, Irshad H, Kuehl PJ, Holmes TD, Sherwood R, Hobbs CH. Lung deposition of droplet aerosols in monkeys. Inhal Toxicol. 2008;20(11):1029–36.

    CAS  Article  Google Scholar 

  4. 4.

    Miller FJ. Dosimeter of particles in laboratory animals and humans in relationship to issues surrounding lung overload and human health risk assessment: a critical review. Inhal Toxicol. 2000;12:19–57.

    CAS  Article  Google Scholar 

  5. 5.

    Alexander DJ, Collins CJ, Coombs DW, Gilkison IS, Hardy CJ, Healey G, Karantabias G, Johnson N, Karlsson A, Kilgour JD, .McDonald P. 'Association of Inhalation Toxicologists (AIT) working party recommendation for standard delivered dose calculation and expression in non-clinical aerosol inhalation toxicology studies with pharmaceuticals', Inhal Toxicol 2008;20(13):1179–1189.

    CAS  Article  Google Scholar 

  6. 6.

    Belser JA, Jacqueline MK, Tumpey TM. The ferret as a model organism to study influenza A virus infection. Dis Model Mech. 2011;4:575–9.

    CAS  Article  Google Scholar 

  7. 7.

    MacInnes H, Zhou Y, Gouveia K, Cromwell J, Lowery L, Layton RC, et al. Transmission of aerosolized seasonal H1N1 influenza A to ferrets. PLoS One. 2011;6(9):e24448.

    CAS  Article  Google Scholar 

  8. 8.

    Lednicky JA, Hamilton SB, Tuttle RS, Sosna WA, Daniels DE, Swayne DE. Ferrets develop fatal influenza after inhaling small particle aerosols of highly pathogenic avian influenza virus A/Vietnam/1203/2004 (H5N1). Virol J. 2010;7:231.

    Article  Google Scholar 

  9. 9.

    Ziying Y, Steward ZA, Sinn PL, Olsen JC, Hu J, McCray PB, et al. Ferret and pig models of cystic fibrosis: prospects and promise for gene therapy. Hum Gene Ther Clin Dev. 2015;26(1):38–49.

    Article  Google Scholar 

  10. 10.

    Ding YO, Hurt AC. Using the ferret as an animal model for investigating influenza antiviral effectiveness. Front Microbiol. 2016;7:80.

    Google Scholar 

  11. 11.

    Panozzo J, Oh DY, Margo K, Morton DA, Piedrafita D, Mosse J, et al. Evaluation of a dry powder delivery system for laninamivir in a ferret model of influenza infection. Antivir Res. 2015;120:66–71.

    CAS  Article  Google Scholar 

  12. 12.

    Hsu J, Santesso N, Mustafa R, Brozek J, Chen YL, Hopkins JP, et al. Antivirals for treatment of influenza: a systematic review and meta-analysis of observational studies. Ann Intern Med. 2012;157(7):512–24.

    Article  Google Scholar 

  13. 13.

    Corcoran TE, Devadason SG, Kuehl PJ. Introduction: aerosol delivery of orally inhaled agents. J Aerosol Med Pulm Drug Deliv. 2012;25(Suppl 1):S3–5.

    Article  Google Scholar 

  14. 14.

    Devadason SG, Chan HK, Haussermann S, Kietzig C, Kuehl PJ, Newman S, et al. Standardization of techniques for validation of radiolabeling of drug formulations for aerosol deposition assessment of orally inhaled products. J Aerosol Med Pulm Drug Deliv. 2012;25(Suppl 1):S6–9.

    Article  Google Scholar 

  15. 15.

    Tepper JS, Kuehl PJ, Cracknell S, Nikula KJ, Pei L, Blanchard JD. Symposium summary: “breathe in, breath out, its easy: what you need to know about developing inhaled drugs”. Int J Toxicol. 2016;35(4):376–92.

    CAS  Article  Google Scholar 

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The authors would like to acknowledge the University of New Mexico Center for Isotopes in Medicine, specifically Tamara Anderson and Jeff Norenberg, for assistance with image acquisition and analysis.

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Correspondence to Philip J. Kuehl.

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David L. Hava and Wesley H. DeHaan were employees of Pulmatrix and held an interest in commercializing products based on aspects of this work.

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Guest Editors: Philip J. Kuehl and Stephen W. Stein

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Kuehl, P.J., Chand, R., McDonald, J.D. et al. Pulmonary and Regional Deposition of Nebulized and Dry Powder Aerosols in Ferrets. AAPS PharmSciTech 20, 242 (2019). https://doi.org/10.1208/s12249-019-1382-3

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  • radiolabeling
  • ferret
  • dry powder inhalation
  • nebulizer inhalation