title: Co-inhalation of roflumilast, rather than formoterol, with fluticasone more effectively improves asthma in asthmatic mice.
abstract: Roflumilast is approved as an add-on therapy for chronic obstructive pulmonary disease. The inflammation in chronic obstructive pulmonary disease is mainly neutrophilic, while in asthma it is mainly eosinophilic, studies addressing role of roflumilast in eosinophilic inflammation are recommended. Also in severe asthma, the dominant inflammatory cells are neutrophils. Thus, roflumilast has a potential off-label use in the treatment of asthma. This study was designed to evaluate the effects of co-inhalation of roflumilast and fluticasone compared to that of formoterol and fluticasone in ovalbumin-sensitized and-challenged BALB/c mice. Besides normal control group, the ovalbumin-asthmatic mice were randomly divided into seven groups (n = 8): positive control, vehicle-treated, and five drug-treated groups. Treatments (µg/kg) were given as 15 min-inhalation once/day for five days as follows: roflumilast (500), formoterol (50), fluticasone (1000), roflumilast + fluticasone (500 + 1000), and formoterol + fluticasone (50 + 1000). Penh values were measured in conscious unrestrained mice using the single-chamber whole-body plethysmography. Airway hyperreactivity to inhaled methacholine was evaluated. Bronchoalveolar lavage fluid was used for the measurements of levels of IL-4, IL-5, TNF-α, OVA-specific IgE, and total and differential white cells. Lung sections were stained with hematoxylin and eosin and periodic acid-Schiff. The asthmatic mice showed significant increases in airway hyperreactivity which were significantly reversed by the combination treatments. The asthmatic mice showed significant increases in levels of IL-4, IL-5, TNF-α, ovalbumin-specific IgE, and total and differential white cells in bronchoalveolar lavage fluid. All treatments (except formoterol) significantly reversed these changes mainly with roflumilast + fluticasone. The asthmatic mice showed severe inflammatory infiltration and goblet cell hyperplasia which were maximally reversed by roflumilast + fluticasone, while minimally reversed by formoterol. In conclusion, co-inhalation of roflumilast + fluticasone more significantly improved inflammation and histopathological changes than co-inhalation of formoterol + fluticasone in ovalumin-asthmatic mice. Further studies are needed to help confirm the potential off-label add-on use of roflumilast in typical and atypical asthma and asthma-chronic obstructive pulmonary disease overlap syndrome. Impact statement Roflumilast, a selective phosphodiesterase-4 inhibitor, was approved for the treatment of chronic obstructive pulmonary disease (COPD). This study showed that co-inhalation of roflumilast and fluticasone significantly decreased airway hyperresponsiveness in ovalumin-asthmatic mice. Also, it more significantly improved inflammation and histopathological changes than co-inhalation of formoterol and fluticasone. The current results showed that inhaled roflumilast reduced counts of eosinophils, neutrophils, and macrophages in bronchoalveolar lavage fluid. Consequently, inhaled roflumilast might be of potential off-label benefit in treatment of eosinophilic and neutrophilic asthma and asthma-COPD overlap syndrome (ACOS). These results could also support other experimental and clinical studies addressing the same issue.
authors.affiliation: 1 Department of Pharmacology, Faculty of Medicine, Rabigh, King Abdulaziz University (KAU), Jeddah 21589, Saudi Arabia.|2 Department of Pharmacology, Faculty of Medicine, Ain Shams University, Cairo 11562, Egypt.
authors.forename: Hussam A
authors.initials: HA
authors.lastname: Murad
authors.affiliation: 3 Department of Pediatrics, Faculty of Medicine, KAU, Jeddah 21589, Saudi Arabia.
authors.forename: Hamed S
authors.initials: HS
authors.lastname: Habib
authors.affiliation: 1 Department of Pharmacology, Faculty of Medicine, Rabigh, King Abdulaziz University (KAU), Jeddah 21589, Saudi Arabia.
authors.forename: Misbahuddin M
authors.initials: MM
authors.lastname: Rafeeq
authors.affiliation: 4 Department of Pharmacology, Faculty of Medicine, KAU, Jeddah 21589, Saudi Arabia.
authors.forename: Mansour I
authors.initials: MI
authors.lastname: Sulaiman
authors.affiliation: 5 Department of Pathology, Faculty of Medicine, Rabigh, KAU, Jeddah 21589, Saudi Arabia.
authors.forename: Amer S
authors.initials: AS
authors.lastname: Abdulrahman
authors.affiliation: 5 Department of Pathology, Faculty of Medicine, Rabigh, KAU, Jeddah 21589, Saudi Arabia.
authors.forename: Mohamad Nidal
authors.initials: MN
authors.lastname: Khabaz
mesh_terms: D000280:Administration, Inhalation; D000631:Aminopyridines / Q000008:administration & dosage / Q000627:therapeutic use*; D000818:Animals; D018927:Anti-Asthmatic Agents / Q000008:administration & dosage / Q000627:therapeutic use*; D001249:Asthma / Q000188:drug therapy*; D001549:Benzamides / Q000008:administration & dosage / Q000627:therapeutic use*; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D003521:Cyclopropanes / Q000008:administration & dosage / Q000627:therapeutic use; D004195:Disease Models, Animal; D004359:Drug Therapy, Combination; D005260:Female; D000068298:Fluticasone / Q000008:administration & dosage / Q000627:therapeutic use*; D000068759:Formoterol Fumarate / Q000008:administration & dosage / Q000627:therapeutic use*; D007073:Immunoglobulin E / Q000032:analysis; D015847:Interleukin-4 / Q000032:analysis; D015848:Interleukin-5 / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis
chemical_list: D000631:Aminopyridines; D018927:Anti-Asthmatic Agents; D001549:Benzamides; D003521:Cyclopropanes; D015848:Interleukin-5; D014409:Tumor Necrosis Factor-alpha; C424423:Roflumilast; D015847:Interleukin-4; D007073:Immunoglobulin E; D000068298:Fluticasone; D000068759:Formoterol Fumarate
keywords: Asthma; fluticasone; formoterol; mice; off-label; ovalbumin; roflumilast