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Feasibility of Tissue Plasminogen Activator Formulated for Pulmonary Delivery

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Purpose

This study was conducted to assess the feasibility of a pulmonary formulation of tissue plasminogen activator (tPA) for nebulization into the airway by measuring protein stability, biologic activity, particle size, and estimating human lung distribution.

Methods

Formulations were derived by varying the surfactant and protein concentrations. Protein stability and recovery of each nebulized tPA formulation were assessed by ultraviolet spectroscopy. Formulations that met protein stability feasibility criteria were assessed for biologic and fibrinolytic activities. Biologic activity was determined by their ability to inhibit superoxide anion production by human neutrophils. Fibrinolytic activity was assessed by the cleavage of plasminogen to plasmin. Aerodynamic properties were assessed using a cascade impactor, and an estimation of human airway deposition was made via a human lung replica.

Results

Twenty-seven tPA formulations were initially assessed, 15 of which met protein stability criteria. Subsequently, three of these formulations maintained biologic and fibrinolytic activities. These formulations exhibited particle sizes of 2.4–3.1 μm, and had respirable doses ≥65%. A formulation of 1mg mL−1 tPA and 0.1% Tween 80 exhibited a 45% deposition in the lower airways of a human lung replica.

Conclusions

A suitable pulmonary tPA formulation was identified that, following nebulization, maintained protein stability as well as biologic and fibrinolytic activities, and resulted in an optimal respirable dose and human airway deposition. This formulation may be applicable in the treatment of lung diseases, such as acute respiratory distress syndrome by permitting targeted pulmonary delivery of a therapeutic protein to the lungs.

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Acknowledgment

This work was funded by a National Institutes of Health Grant R41 HL071439-01A1.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, School of Pharmacy, University of Colorado Health Sciences Center, Denver, Colorado, USA

    John S. Dunn

  2. HTD Biosystems, Inc., Hercules, California, USA

    Rajiv Nayar & Jackie Campos

  3. Webb Waring Institute for Antioxidant Research, University of Colorado Health Sciences Center, Denver, Colorado, USA

    Brooks M. Hybertson & John E. Repine

  4. Lovelace Respiratory Research Institute, Albuquerque, New Mexico, USA

    Yue Zhou

  5. Legacy BioDesign LLC, Loveland, Colorado, USA

    Mark Cornell Manning

  6. Department of Clinical Pharmacy, School of Pharmacy, University of Colorado Health Sciences Center, C238, 4200 East Ninth Avenue, Denver, Colorado, 80262, USA

    Kathleen A. Stringer

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  1. John S. Dunn
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  2. Rajiv Nayar
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  3. Jackie Campos
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  4. Brooks M. Hybertson
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  7. John E. Repine
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  8. Kathleen A. Stringer
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Correspondence to Kathleen A. Stringer.

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Dunn, J.S., Nayar, R., Campos, J. et al. Feasibility of Tissue Plasminogen Activator Formulated for Pulmonary Delivery. Pharm Res 22, 1700–1707 (2005). https://doi.org/10.1007/s11095-005-6335-8

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  • DOI: https://doi.org/10.1007/s11095-005-6335-8

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