title: Toward Repositioning Niclosamide for Antivirulence Therapy of Pseudomonas aeruginosa Lung Infections: Development of Inhalable Formulations through Nanosuspension Technology.
abstract: Inhaled antivirulence drugs are currently considered a promising therapeutic option to treat Pseudomonas aeruginosa lung infections in cystic fibrosis (CF). We have recently shown that the anthelmintic drug niclosamide (NCL) has strong quorum sensing (QS) inhibiting activity against P. aeruginosa and could be repurposed as an antivirulence drug. In this work, we developed dry powders containing NCL nanoparticles that can be reconstituted in saline solution to produce inhalable nanosuspensions. NCL nanoparticles were produced by high-pressure homogenization (HPH) using polysorbate 20 or polysorbate 80 as stabilizers. After 20 cycles of HPH, all formulations showed similar properties in the form of needle-shape nanocrystals with a hydrodynamic diameter of approximately 450 nm and a zeta potential of -20 mV. Nanosuspensions stabilized with polysorbate 80 at 10% w/w to NCL (T80_10) showed an optimal solubility profile in simulated interstitial lung fluid. T80_10 was successfully dried into mannitol-based dry powder by spray drying. Dry powder (T80_10 DP) was reconstituted in saline solution and showed optimal in vitro aerosol performance. Both T80_10 and T80_10 DP were able to inhibit P. aeruginosa QS at NCL concentrations of 2.5-10 μM. NCL, and these formulations did not significantly affect the viability of CF bronchial epithelial cells in vitro at microbiologically active concentrations (i.e., ≤10 μM). In vivo acute toxicity studies in rats confirmed no observable toxicity of the NCL T80_10 DP formulation upon intratracheal administration at a concentration 100-fold higher than the anti-QS activity concentration. These preliminary results suggest that NCL repurposed in the form of inhalable nanosuspensions has great potential for the local treatment of P. aeruginosa lung infections as in the case of CF patients.
authors.affiliation: †Department of Pharmacy, University of Naples Federico II, Via Domenico Montesano 49, 80131 Naples, Italy.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Costabile
authors.affiliation: ‡Di.S.T.A.Bi.F., Second University of Naples, Via Vivaldi 43, 81100 Caserta, Italy.
authors.forename: Ivana
authors.initials: I
authors.lastname: d'Angelo
authors.affiliation: §Department of Sciences, University Roma Tre, Viale Marconi, 446, 00146 Rome, Italy.
authors.forename: Giordano
authors.initials: G
authors.lastname: Rampioni
authors.affiliation: §Department of Sciences, University Roma Tre, Viale Marconi, 446, 00146 Rome, Italy.
authors.forename: Roslen
authors.initials: R
authors.lastname: Bondì
authors.affiliation: ⊥Department of Biology and Biotechnology "Charles Darwin", Sapienza University of Rome, Via dei Sardi 70, 00185 Rome, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Pompili
authors.affiliation: ⊥Department of Biology and Biotechnology "Charles Darwin", Sapienza University of Rome, Via dei Sardi 70, 00185 Rome, Italy.
authors.forename: Fiorentina
authors.initials: F
authors.lastname: Ascenzioni
authors.affiliation: †Department of Pharmacy, University of Naples Federico II, Via Domenico Montesano 49, 80131 Naples, Italy.
authors.forename: Emma
authors.initials: E
authors.lastname: Mitidieri
authors.affiliation: †Department of Pharmacy, University of Naples Federico II, Via Domenico Montesano 49, 80131 Naples, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: d'Emmanuele di Villa Bianca
authors.affiliation: †Department of Pharmacy, University of Naples Federico II, Via Domenico Montesano 49, 80131 Naples, Italy.
authors.forename: Raffaella
authors.initials: R
authors.lastname: Sorrentino
authors.affiliation: †Department of Pharmacy, University of Naples Federico II, Via Domenico Montesano 49, 80131 Naples, Italy.
authors.forename: Agnese
authors.initials: A
authors.lastname: Miro
authors.affiliation: †Department of Pharmacy, University of Naples Federico II, Via Domenico Montesano 49, 80131 Naples, Italy.
authors.forename: Fabiana
authors.initials: F
authors.lastname: Quaglia
authors.affiliation: ⊥Department of Biology and Biotechnology "Charles Darwin", Sapienza University of Rome, Via dei Sardi 70, 00185 Rome, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Imperi
authors.affiliation: §Department of Sciences, University Roma Tre, Viale Marconi, 446, 00146 Rome, Italy.
authors.forename: Livia
authors.initials: L
authors.lastname: Leoni
authors.affiliation: †Department of Pharmacy, University of Naples Federico II, Via Domenico Montesano 49, 80131 Naples, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Ungaro
mesh_terms: D000280:Administration, Inhalation; D000818:Animals; D000900:Anti-Bacterial Agents / Q000008:administration & dosage* / Q000737:chemistry; D002626:Chemistry, Pharmaceutical; D004353:Drug Evaluation, Preclinical; D058492:Drug Repositioning / Q000639:trends; D008171:Lung Diseases / Q000188:drug therapy* / Q000382:microbiology / Q000473:pathology; D008297:Male; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry; D009534:Niclosamide / Q000008:administration & dosage* / Q000737:chemistry; D011208:Powders; D011552:Pseudomonas Infections / Q000188:drug therapy* / Q000473:pathology; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000472:pathogenicity; D051381:Rats; D017208:Rats, Wistar; D014774:Virulence / Q000187:drug effects
chemical_list: D000900:Anti-Bacterial Agents; D011208:Powders; D009534:Niclosamide
keywords: Pseudomonas aeruginosa; antivirulence drugs; cystic fibrosis; nanosuspensions; niclosamide; pulmonary delivery; quorum sensing; rat