Skip to main content

Advertisement

SpringerLink
Log in

A Dry Powder Formulation of Liposome-Encapsulated Recombinant Secretory Leukocyte Protease Inhibitor (rSLPI) for Inhalation: Preparation and Characterisation

  • Research Article
  • Published:
AAPS PharmSciTech Aims and scope Submit manuscript

Abstract

Inhaled recombinant secretory leukocyte protease inhibitor (rSLPI) has shown potential for the treatment of inflammatory lung conditions. Rapid inactivation of rSLPI by cathepsin L (Cat L) and rapid clearance from the lungs has limited clinical efficacy to date. Previous studies by us have shown that encapsulation of rSLPI within1,2-dioleoyl-sn-glycero-3-[phospho-L-serine]/cholesterol (DOPS/Chol) liposomes protects rSLPI against Cat L inactivation in vitro. Liquid DOPS–rSLPI preparations were found to be unstable upon long-term storage and nebulisation. The aim of this study was therefore to develop a method of manufacture for preparing DOPS–rSLPI liposomes as a dry powder for inhalation. DOPS–rSLPI dry powders were lyophilised and subsequently micronised with a novel micronisation aid. The effects of formulation and processing on rSLPI stability, activity, and uniformity of content within the powders were characterised. Using D-mannitol as the micronisation aid, dry powder particles in the inhalable size range (<5 μm) were prepared. By optimising process parameters, up to 54% of rSLPI was recovered after micronisation, of which there was no significant loss in anti-neutrophil elastase activity and no detectable evidence of protein degradation. Aerosolisation was achieved using a dry powder inhaler, and mass median aerodynamic diameter (MMAD) was evaluated after collection in a cascade impactor. Aerosolisation of the DOPS–rSLPI dry powder yielded 38% emitted dose, with 2.44 μm MMAD. When challenged with Cat L post-aerosolisation, DOPS–rSLPI dry powder was significantly better at retaining a protective function against Cat L-induced rSLPI inactivation compared to the aqueous DOPS–rSLPI liposome dispersion and was also more stable under storage.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

ACI:

Andersen cascade impactor

Anti-NE:

Anti-neutrophil elastase

Cat L:

Cathepsin L

CFCs:

Chlorofluorocarbons

Chol:

Cholesterol

DOPS:

1,2-Dioleoyl-sn-glycero-3-[phospho-L-serine]

DOPS–rSLPI:

rSLPI encapsulated in DOPS/Chol liposomes

DPI:

Dry powder inhaler

ED:

Emitted dose

FPF:

Fine particle fraction

GSD:

Geometric standard deviation

MMAD:

Mass median aerodynamic diameter

NE:

Neutrophil elastase

NRF:

Non-respirable fraction

SEM:

Scanning electron microscopy

rSLPI:

Recombinant secretory leukocyte protease inhibitor

SLPI:

Secretory leukocyte protease inhibitor

TSI:

Twin stage impinger

References

  1. Barrios VE, Middleton SC, Kashem MA, Havill AM, Toombs CF, Wright CD. Tryptase mediates hyperresponsiveness in isolated guinea pig bronchi. Life Sci. 1998;63(26):2295–303.

    Article  PubMed  CAS  Google Scholar 

  2. Forteza RM, Ahmed A, Lee T, Abraham WM. Secretory leukocyte protease inhibitor, but not alpha-1 protease inhibitor, blocks tryptase-induced bronchoconstriction. Pulm Pharmacol Ther. 2001;14(2):107–10.

    Article  PubMed  CAS  Google Scholar 

  3. Gillissen A, Birrer P, McElvaney NG, Buhl R, Vogelmeier C, Hoyt Jr RF, et al. Recombinant secretory leukoprotease inhibitor augments glutathione levels in lung epithelial lining fluid. J Appl Physiol. 1993;75(2):825–32.

    PubMed  CAS  Google Scholar 

  4. McElvaney NG, Doujaiji B, Moan MJ, Burnham MR, Wu MC, Crystal RG. Pharmacokinetics of recombinant secretory leukoprotease inhibitor aerosolized to normals and individuals with cystic fibrosis. Am Rev Respir Dis. 1993;148(4 Pt 1):1056–60.

    PubMed  CAS  Google Scholar 

  5. Wright CD, Havill AM, Middleton SC, Kashem MA, Lee PA, Dripps DJ, et al. Secretory leukocyte protease inhibitor prevents allergen-induced pulmonary responses in animal models of asthma. J Pharmacol Exp Ther. 1999;289(2):1007–14.

    PubMed  CAS  Google Scholar 

  6. Keatings VM, Jatakanon A, Worsdell YM, Barnes PJ. Effects of inhaled and oral glucocorticoids on inflammatory indices in asthma and COPD. Am J Respir Crit Care Med. 1997;155(2):542–8.

    PubMed  CAS  Google Scholar 

  7. Adcock IM. Steroid resistance in asthma. Molecular mechanisms. Am J Respir Crit Care Med. 1996;154(2 Pt 2):S58–61.

    PubMed  CAS  Google Scholar 

  8. Bergenfeldt M, Bjork P, Ohlsson K. The elimination of secretory leukocyte protease inhibitor (SLPI) after intravenous injection in dog and man. Scand J Clin Lab Invest. 1990;50(7):729–37.

    Article  PubMed  CAS  Google Scholar 

  9. Stolk J, Camps J, Feitsma HI, Hermans J, Dijkman JH, Pauwels EK. Pulmonary deposition and disappearance of aerosolised secretory leucocyte protease inhibitor. Thorax. 1995;50(6):645–50.

    Article  PubMed  CAS  Google Scholar 

  10. Gast A, Anderson W, Probst A, Nick H, Thompson RC, Eisenberg SP, et al. Pharmacokinetics and distribution of recombinant secretory leukocyte proteinase inhibitor in rats. Am Rev Respir Dis. 1990;141(4 Pt 1):889–94.

    PubMed  CAS  Google Scholar 

  11. McElvaney NG, Nakamura H, Birrer P, Hebert CA, Wong WL, Alphonso M, et al. Modulation of airway inflammation in cystic fibrosis. In vivo suppression of interleukin-8 levels on the respiratory epithelial surface by aerosolization of recombinant secretory leukoprotease inhibitor. J Clin Invest. 1992;90(4):1296–301.

    Article  PubMed  CAS  Google Scholar 

  12. Vogelmeier C, Gillissen A, Buhl R. Use of secretory leukoprotease inhibitor to augment lung antineutrophil elastase activity. Chest. 1996;110(6 Suppl):261S–6S.

    Article  PubMed  CAS  Google Scholar 

  13. Taggart CC, Lowe GJ, Greene CM, Mulgrew AT, O’Neill SJ, Levine RL, et al. Cathepsin B, L, and S cleave and inactivate secretory leucoprotease inhibitor. J Biol Chem. 2001;276(36):33345–52.

    Article  PubMed  CAS  Google Scholar 

  14. Couvreur P, Fattal E, Andremont A. Liposomes and nanoparticles in the treatment of intracellular bacterial infections. Pharm Res. 1991;8(9):1079–86.

    Article  PubMed  CAS  Google Scholar 

  15. McCullough HN, Juliano RL. Organ-selective action of an antitumor drug: pharmacologic studies of liposome-encapsulated beta-cytosine arabinoside administered via the respiratory system of the rat. J Natl Cancer Inst. 1979;63(3):727–31.

    PubMed  CAS  Google Scholar 

  16. Taylor KM, Taylor G, Kellaway IW, Stevens J. The influence of liposomal encapsulation on sodium cromoglycate pharmacokinetics in man. Pharm Res. 1989;6(7):633–6.

    Article  PubMed  CAS  Google Scholar 

  17. Niven RW, Schreier H. Nebulization of liposomes. I. Effects of lipid composition. Pharm Res. 1990;7(11):1127–33.

    Article  PubMed  CAS  Google Scholar 

  18. Misra A, Jinturkar K, Patel D, Lalani J, Chougule M. Recent advances in liposomal dry powder formulations: preparation and evaluation. Expert Opin Drug Deliv. 2009;6(1):71–89.

    Article  PubMed  CAS  Google Scholar 

  19. Clark JM, Whitney RR, Olsen SJ, George RJ, Swerdel MR, Kunselman L, et al. Amphotericin B lipid complex therapy of experimental fungal infections in mice. Antimicrob Agents Chemother. 1991;35(4):615–21.

    PubMed  CAS  Google Scholar 

  20. Freise CE, Liu T, Hong K, Osorio RW, Papahadjopoulos D, Ferrell L, et al. The increased efficacy and decreased nephrotoxicity of a cyclosporine liposome. Transplantation. 1994;57(6):928–32.

    Article  PubMed  CAS  Google Scholar 

  21. Gruber SA, Venkataram S, Canafax DM, Cipolle RJ, Bowers L, Elsberry D, et al. Liposomal formulation eliminates acute toxicity and pump incompatibility of parenteral cyclosporine. Pharm Res. 1989;6(7):601–7.

    Article  PubMed  CAS  Google Scholar 

  22. Wyde PR, Six HR, Wilson SZ, Gilbert BE, Knight V. Activity against rhinoviruses, toxicity, and delivery in aerosol of enviroxime in liposomes. Antimicrob Agents Chemother. 1988;32(6):890–5.

    PubMed  CAS  Google Scholar 

  23. Bhavane R, Karathanasis E, Annapragada AV. Agglomerated vesicle technology: a new class of particles for controlled and modulated pulmonary drug delivery. J Control Release. 2003;93(1):15–28.

    Article  PubMed  CAS  Google Scholar 

  24. Gibbons AM, McElvaney NG, Taggart CC, Cryan SA. Delivery of rSLPI in a liposomal carrier for inhalation provides protection against cathepsin L degradation. J Microencapsul. 2009;26:513–22.

    Article  PubMed  CAS  Google Scholar 

  25. Hansen NC, Evald T, Ibsen TB. Terbutaline inhalations by the Turbuhaler as replacement for domiciliary nebulizer therapy in severe chronic obstructive pulmonary disease. Respir Med. 1994;88(4):267–71.

    Article  PubMed  CAS  Google Scholar 

  26. Chougule MB, Padhi BK, Jinturkar KA, Misra A. Development of dry powder inhalers. Recent Pat Drug Deliv Formul. 2007;1:11–21.

    Article  PubMed  CAS  Google Scholar 

  27. Zhu J, Wen J, Ma Y, Zhang H, Zhu J, Wen J, Ma Y, Zhang H, Zhu J, Wen J, Ma Y, Zhang Hs. Dry Powder Inhaler. USA. 2007.

  28. Newman SP. Dry powder inhalers for optimal drug delivery. Expert Opin Biol Ther. 2004;4(1):23–33.

    Article  PubMed  CAS  Google Scholar 

  29. Niven RW. Delivery of biotherapeutics by inhalation aerosol. Crit Rev Ther Drug Carrier Syst. 1995;12(2–3):151–231.

    PubMed  CAS  Google Scholar 

  30. Salnikova MS, Middaugh CR, Rytting JH. Stability of lyophilized human growth hormone. Int J Pharm. 2008;358(1–2):108–13.

    Article  PubMed  CAS  Google Scholar 

  31. DePaz RA, Dale DA, Barnett CC, Carpenter JF, Gaertner AL, Randolph TW. Effects of drying methods and additives on the structure, function, and storage stability of subtilisin: role of protein conformation and molecular mobility. Enzyme Microb Technol. 2002;31(6):765–74.

    Article  CAS  Google Scholar 

  32. Franks F. Freeze-drying of bioproducts: putting principles into practice. Eur J Pharm Biopharm. 1998;45(3):221–9.

    Article  PubMed  CAS  Google Scholar 

  33. Kensil CR, Dennis EA. Alkaline hydrolysis of phospholipids in model membranes and the dependence on their state of aggregation. Biochemistry. 1981;20(21):6079–85.

    Article  PubMed  CAS  Google Scholar 

  34. Grit M, Zuidam NJ, Underberg WJ, Crommelin DJ. Hydrolysis of partially saturated egg phosphatidylcholine in aqueous liposome dispersions and the effect of cholesterol incorporation on hydrolysis kinetics. J Pharm Pharmacol. 1993;45(6):490–5.

    PubMed  CAS  Google Scholar 

  35. Grit M, Crommelin DJ. The effect of aging on the physical stability of liposome dispersions. Chem Phys Lipids. 1992;62(2):113–22.

    Article  PubMed  CAS  Google Scholar 

  36. Vemuri S, Rhodes CT. Preparation and characterization of liposomes as therapeutic delivery systems: a review. Pharm Acta Helv. 1995;70(2):95–111.

    Article  PubMed  CAS  Google Scholar 

  37. Ausborn M, Nuhn P, Schreier H. Stabilization of liposomes by freeze-thaw- and lyophilization techniques: problems and opportunities. Eur J Pharm Biopharm. 1992;38(4):133–8.

    CAS  Google Scholar 

  38. Lloyd AW, Olliff CJ, Rutt KJ. A comparison of carboxylate salts as liposomal cryoprotectants. Int J Pharm. 1996;131(2):257–62.

    Article  CAS  Google Scholar 

  39. Glavas-Dodov M, Fredro-Kumbaradzi E, Goracinova K, Simonoska M, Calis S, Trajkovic-Jolevska S, et al. The effects of lyophilization on the stability of liposomes containing 5-FU. Int J Pharm. 2005;291(1–2):79–86.

    Article  PubMed  CAS  Google Scholar 

  40. Zhang JA, Xuan T, Parmar M, Ma L, Ugwu S, Ali S, et al. Development and characterization of a novel liposome-based formulation of SN-38. Int J Pharm. 2004;270(1–2):93–107.

    Article  PubMed  CAS  Google Scholar 

  41. Shulkin PM, Seltzer SE, Davis MA, Adams DF. Lyophilized liposomes: a new method for long-term vesicular storage. J Microencapsul. 1984;1(1):73–80.

    Article  PubMed  CAS  Google Scholar 

  42. Changsan N, Chan HK, Separovic F, Srichana T. Physicochemical characterization and stability of rifampicin liposome dry powder formulations for inhalation. J Pharm Sci. 2009;98:628–39.

    Article  PubMed  CAS  Google Scholar 

  43. van Bommel EMG, Crommelin DJ. Stability of doxorubicin-liposomes on storage: as an aqueous dispersion, frozen or freeze-dried. Int J Pharm. 1984;22:299–310.

    Article  Google Scholar 

  44. Taylor KMG, Taylor G, Kellaway IW, Stevens J. The stability of liposomes to nebulisation. Int J Pharm. 1990;58(1):57–61.

    Article  CAS  Google Scholar 

  45. Niven RW, Speer M, Schreier H. Nebulization of liposomes. II. The effects of size and modeling of solute release profiles. Pharm Res. 1991;8(2):217–21.

    Article  PubMed  CAS  Google Scholar 

  46. Bridges PA, Taylor AJ, McCallion ON. Nebulisation of liposomes: the effect of formulation variables. Bristol: The Aerosol Society; 1995.

    Google Scholar 

  47. Bridges PA, Taylor AJ, McCallion ON. The effects of lipid concentration on the nebulisation of liposomes. Proc Int Symp Control Release Bioact Mater. 1995;22:2021–2.

    Google Scholar 

  48. McCallion ON, Taylor KM, Thomas M, Taylor AJ. Nebulization of fluids of different physicochemical properties with air-jet and ultrasonic nebulizers. Pharm Res. 1995;12(11):1682–8.

    Article  PubMed  CAS  Google Scholar 

  49. Stewart JC. Colorimetric determination of phospholipids with ammonium ferrothiocyanate. Anal Biochem. 1980;104(1):10–4.

    Article  PubMed  CAS  Google Scholar 

  50. Mayer LD, Bally MB, Hope MJ, Cullis PR. Techniques for encapsulating bioactive agents into liposomes. Chem Phys Lipids. 1986;40(2–4):333–45.

    Article  PubMed  CAS  Google Scholar 

  51. Niven RW, Carvajal TM, Schreier H. Nebulization of liposomes. III. The effects of operating conditions and local environment. Pharm Res. 1992;9(4):515–20.

    Article  PubMed  CAS  Google Scholar 

  52. Patton JS, Bukar J, Nagarajan S. Inhaled insulin. Adv Drug Deliv Rev. 1999;35(2–3):235–47.

    Article  PubMed  CAS  Google Scholar 

  53. Rave KM, Nosek L, de la Pena A, Seger M, Ernest 2nd CS, Heinemann L, et al. Dose response of inhaled dry-powder insulin and dose equivalence to subcutaneous insulin lispro. Diab Care. 2005;28(10):2400–5.

    Article  CAS  Google Scholar 

  54. Hollander PA, Blonde L, Rowe R, Mehta AE, Milburn JL, Hershon KS, et al. Efficacy and safety of inhaled insulin (exubera) compared with subcutaneous insulin therapy in patients with type 2 diabetes: results of a 6-month, randomized, comparative trial. Diab Care. 2004;27(10):2356–62.

    Article  CAS  Google Scholar 

  55. Codrons V, Vanderbist F, Verbeeck RK, Arras M, Lison D, Preat V, et al. Systemic delivery of parathyroid hormone (1–34) using inhalation dry powders in rats. J Pharm Sci. 2003;92(5):938–50.

    Article  PubMed  CAS  Google Scholar 

  56. Schreier H, Mobley WC, Concessio N, Hickey AJ, Niven R. Formulation and in vitro performance of liposome powder aerosol. STP Pharm Sci. 1994;4:38–44.

    CAS  Google Scholar 

  57. Mobley WC. The effect of jet-milling on lyophilized liposomes. Pharm Res. 1998;15(1):149–52.

    Article  PubMed  CAS  Google Scholar 

  58. Bloom M, Evans E, Mouritsen OG. Physical properties of the fluid lipid-bilayer component of cell membranes: a perspective. Q Rev Biophys. 1991;24(3):293–397.

    Article  PubMed  CAS  Google Scholar 

  59. Yu L, Mishra DS, Rigsbee DR. Determination of the glass properties of D-mannitol using sorbitol as an impurity. J Pharm Sci. 1998;87(6):774–7.

    Article  PubMed  CAS  Google Scholar 

  60. Desai TR, Hancock RE, Finlay WH. Delivery of liposomes in dry powder form: aerodynamic dispersion properties. Eur J Pharm Sci. 2003;20(4–5):459–67.

    Article  PubMed  CAS  Google Scholar 

  61. Shah SP, Misra A. Liposomal amikacin dry powder inhaler: effect of fines on in vitro performance. AAPS PharmSciTech. 2004;5(4):e65.

    Article  PubMed  Google Scholar 

  62. Desai TR, Wong JP, Hancock RE, Finlay WH. A novel approach to the pulmonary delivery of liposomes in dry powder form to eliminate the deleterious effects of milling. J Pharm Sci. 2002;91(2):482–91.

    Article  PubMed  CAS  Google Scholar 

  63. Jones MD, Price R. The influence of fine excipient particles on the performance of carrier-based dry powder inhalation formulations. Pharm Res. 2006;23(8):1665–74.

    Article  PubMed  CAS  Google Scholar 

  64. Shah SP, Misra A. Liposomal amphotericin B dry powder inhaler: effect of fines on in vitro performance. Pharmazie. 2004;59(10):812–3.

    PubMed  CAS  Google Scholar 

  65. Lu D, Hickey AJ. Liposomal dry powders as aerosols for pulmonary delivery of proteins. AAPS PharmSciTech. 2005;6(4):E641–8.

    Article  PubMed  Google Scholar 

  66. Joshi M, Misra A. Dry powder inhalation of liposomal Ketotifen fumarate: formulation and characterization. Int J Pharm. 2001;223(1–2):15–27.

    Article  PubMed  CAS  Google Scholar 

  67. Joshi MR, Misra A. Liposomal budesonide for dry powder inhaler: preparation and stabilization. AAPS PharmSciTech. 2001;2(4):25.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This study was funded by the Irish Research Council for Science, Engineering and Technology (SC/2004/B0419) and the Health Research Board (HRB). The authors gratefully acknowledge the supply of rSLPI from Amgen®.

Author information

Authors and Affiliations

  1. School of Pharmacy, Royal College of Surgeons in Ireland, York House, York Street, Dublin 2, Ireland

    Aileen Gibbons & Sally-Ann Cryan

  2. Department of Medicine, Beaumont Hospital, Royal College of Surgeons in Ireland, Dublin, Ireland

    Noel G. McElvaney

Authors
  1. Aileen Gibbons
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Noel G. McElvaney
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sally-Ann Cryan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sally-Ann Cryan.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gibbons, A., McElvaney, N.G. & Cryan, SA. A Dry Powder Formulation of Liposome-Encapsulated Recombinant Secretory Leukocyte Protease Inhibitor (rSLPI) for Inhalation: Preparation and Characterisation. AAPS PharmSciTech 11, 1411–1421 (2010). https://doi.org/10.1208/s12249-010-9500-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1208/s12249-010-9500-2

KEY WORDS

Search

Navigation