title: Development of inhalable hyaluronan/mannitol composite dry powders for flucytosine repositioning in local therapy of lung infections.
abstract: Flucytosine (5-fluorocytosine, 5-FC) is a fluorinated analogue of cytosine currently approved for the systemic treatment of fungal infections, which has recently demonstrated a very promising antivirulence activity against the bacterial pathogen Pseudomonas aeruginosa. In this work, we propose novel inhalable hyaluronic acid (HA)/mannitol composite dry powders for repositioning 5-FC in the local treatment of lung infections, including those affecting cystic fibrosis (CF) patients. Different dry powders were produced in one-step by spray-drying. Powder composition and process conditions were selected after in depth formulation studies aimed at selecting the 5-FC/HA/mannitol formulation with convenient aerosolization properties and drug release profile in simulated lung fluids. The optimized 5-FC/HA/mannitol powder for inhalation (HyaMan_FC#3) was effectively delivered from different breath-activated dry powder inhalers (DPI) already available to CF patients. Nevertheless, the aerodynamic assessment of fine particles suggested that the developed formulation well fit with a low-resistance DPI. HyaMan_FC#3 inhibited the growth of the fungus Candida albicans and the production of the virulence factor pyoverdine by P. aeruginosa at 5-FC concentrations that did not affect the viability of both wild type (16HBE14o-) and CF (CFBE41o-) human bronchial epithelial cells. Finally, pharmacokinetics of HyaMan_FC#3 inhalation powder and 5-FC solution after intratracheal administration in rats were compared. In vivo results clearly demonstrated that, when formulated as dry powder, 5-FC levels in both bronchoalveolar lavage fluid and lung tissue were significantly higher and sustained over time as compared to those obtained with the 5-FC solution. Of note, when the same 5-FC amount was administered intravenously, no significant drug amount was found in the lung at each time point from the injection. To realize a 5-FC lung concentration similar to that obtained by using HyaMan_FC#3, a 6-fold higher dose of 5-FC should be administered intravenously. Taken together, our data demonstrate the feasibility to deliver 5-FC by the pulmonary route likely avoiding/reducing the well-known side effects associated to the high systemic 5-FC doses currently used in humans. Furthermore, our results highlight that an appropriate formulation design can improve the persistence of the drug at lungs, where microorganisms causing severe infections are located.
authors.affiliation: Laboratory of Drug Delivery, Department of Pharmacy, University of Napoli Federico II, Via Domenico Montesano 49, 80131 Napoli, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Costabile
authors.affiliation: Di.S.T.A.Bi.F., Second University of Napoli, Via Vivaldi 43, 81100 Caserta, Italy.
authors.forename: I
authors.initials: I
authors.lastname: d'Angelo
authors.affiliation: Laboratory of Pharmacology, Department of Pharmacy, University of Napoli Federico II, Via Domenico Montesano 49, 80131 Napoli, Italy.
authors.forename: R
authors.initials: R
authors.lastname: d'Emmanuele di Villa Bianca
authors.affiliation: Laboratory of Pharmacology, Department of Pharmacy, University of Napoli Federico II, Via Domenico Montesano 49, 80131 Napoli, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Mitidieri
authors.affiliation: Department of Biology and Biotechnology "Charles Darwin", Sapienza University of Roma, Via dei Sardi 70, 00185 Roma, Italy.
authors.forename: B
authors.initials: B
authors.lastname: Pompili
authors.affiliation: Department of Biology and Biotechnology "Charles Darwin", Sapienza University of Roma, Via dei Sardi 70, 00185 Roma, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Del Porto
authors.affiliation: Department of Sciences, University Roma Tre, Viale Marconi, 446, 00146 Roma, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Leoni
authors.affiliation: Department of Sciences, University Roma Tre, Viale Marconi, 446, 00146 Roma, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Visca
authors.affiliation: Laboratory of Drug Delivery, Department of Pharmacy, University of Napoli Federico II, Via Domenico Montesano 49, 80131 Napoli, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Miro
authors.affiliation: Laboratory of Drug Delivery, Department of Pharmacy, University of Napoli Federico II, Via Domenico Montesano 49, 80131 Napoli, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Quaglia
authors.affiliation: Department of Biology and Biotechnology "Charles Darwin", Sapienza University of Roma, Via dei Sardi 70, 00185 Roma, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Imperi
authors.affiliation: Laboratory of Pharmacology, Department of Pharmacy, University of Napoli Federico II, Via Domenico Montesano 49, 80131 Napoli, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Sorrentino
authors.affiliation: Laboratory of Drug Delivery, Department of Pharmacy, University of Napoli Federico II, Via Domenico Montesano 49, 80131 Napoli, Italy. Electronic address: ungaro@unina.it.
authors.forename: F
authors.initials: F
authors.lastname: Ungaro
mesh_terms: D000280:Administration, Inhalation; D000336:Aerosols / Q000737:chemistry; D000818:Animals; D000900:Anti-Bacterial Agents / Q000008:administration & dosage* / Q000493:pharmacokinetics / Q000494:pharmacology; D000935:Antifungal Agents / Q000008:administration & dosage* / Q000493:pharmacokinetics / Q000494:pharmacology; D002176:Candida albicans / Q000187:drug effects; D002177:Candidiasis / Q000188:drug therapy; D058492:Drug Repositioning; D058995:Dry Powder Inhalers; D005437:Flucytosine / Q000008:administration & dosage* / Q000493:pharmacokinetics / Q000494:pharmacology; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry*; D008168:Lung / Q000382:microbiology; D008172:Lung Diseases, Fungal / Q000188:drug therapy; D008297:Male; D008353:Mannitol / Q000737:chemistry*; D010316:Particle Size; D011208:Powders; D011552:Pseudomonas Infections / Q000188:drug therapy; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D017208:Rats, Wistar
chemical_list: D000336:Aerosols; D000900:Anti-Bacterial Agents; D000935:Antifungal Agents; D011208:Powders; D008353:Mannitol; D006820:Hyaluronic Acid; D005437:Flucytosine
keywords: Candida albicans; Flucytosine; Hyaluronic acid; Inhalable powder; Lung delivery; Pseudomonas aeruginosa