Assessment of pulmonary delivery of Plasminogen for ARDS local treatment
Vizzoni1,2, C. Migone1, B. Grassiri1, F. Mori3, R. Crea3, S. Esin4, G. Batoni4, B. Ferro5, A.M. Piras1
1 University of Pisa, Dept. of Pharmacy, Pisa, Italy; 2 University of Siena, Dept. of Life Sciences, Siena, Italy
3 Kedrion S.p.A., Loc. Bolognana, Gallicano, Lucca, Italy; 4 University of Pisa, Dept. of Translational Research and New Technologies in Medicine and Surgery, Pisa, Italy; 5 Spedali Riuniti Livorno, Azienda USL Toscana Nord Ovest, Italy
Summary
Acute respiratory distress syndrome (ARDS) is a severe complication of lung injuries, related to bacterial, fungal and viral infections. ARDS is dramatically correlated to patient mortality and no effective treatment is presently available. ARDS involves hypercoagulation, fibrin deposition in the development of an obstructing hyaline membrane, and deep lung inflammation. Plasminogen (PLG) is component of the fibrinolytic system and contributes to inflammation regulatory processes. Pulmonary delivery of PLG, as an off-label administration of PLG-OMP eye drops (plasminogen-based orphan medicinal product) is here proposed as a readily available opportunity for ARDS treatment. The nebulisation was performed using either jet, ultrasonic or mesh nebulisers. Several investigations were performed on the nebulised samples, such as protein aggregation/fragmentation, PLG glycoform distribution, droplets size and aerodynamic distribution as well as enzymatic activity maintenance. Thanks to the excellent efficacy of the mesh aerosolised samples, additional in vitro studies were performed. Mesh nebulised PLG demonstrated good penetration through artificial airway mucus, low cytotoxicity and poor permeation across an Air-Liquid Interface cellular model of pulmonary epithelium. The results suggest for a good safety profile, excluding high systemic absorption but good mucus diffusion. Most importantly, the aerosolised PLG reversed the effects of LPS-activated macrophage cells, demonstrating an immunomodulating activity of PLG treatment in an already induced inflammatory state. Repeated-dose toxicity studies in mice confirmed the tolerability of PLG-OMP through intra-tracheal administration. In vivo studies on LPS induced ARDS murine model indicate efficacy in avoiding the most important pathological signs of acute ARDS.