Roflumilast N-oxide reverses corticosteroid resistance in neutrophils from patients with chronic obstructive pulmonary disease.

Abstract

BACKGROUND:

Glucocorticoid functions are markedly impaired in patients with chronic obstructive pulmonary disease (COPD). The phosphodiesterase 4 inhibitor roflumilast N-oxide (RNO) is the active metabolite of roflumilast approved as a treatment to reduce the risk of exacerbations in patients with severe COPD.

OBJECTIVE:

We sought to characterize the differential effects of RNO versus corticosteroids and their potential additive/synergistic effect in neutrophils from patients with COPD, thus providing scientific rationale for the combination of roflumilast with corticosteroids in the clinic.

METHODS:

Peripheral blood neutrophils were isolated from patients with COPD (n = 32), smokers (n = 7), and healthy nonsmokers (n = 25). Levels of IL-8, matrix metallopeptidase 9 (MMP-9), and biomarkers of glucocorticoid resistance were determined by using ELISA and RT-PCR. Neutrophils were incubated with dexamethasone (0.1 nmol/L to 1 μmol/L), RNO (0.1 nmol/L to 1 μmol/L), or the combination of 1 nmol/L RNO plus 10 nmol/L DEX and stimulated with LPS (1 μg/mL) or cigarette smoke extract 5%; levels of IL-8, MMP-9, and other biomarkers were measured at the end of the incubation period.

RESULTS:

Peripheral neutrophils from patients with COPD showed a primed phenotype with an increased basal release of IL-8 and MMP-9 and expressed a corticosteroid resistance molecular profile characterized by an increase in phosphoinositide 3-kinase δ, macrophage migration inhibitory factor, and glucocorticoid receptor β expression and a decrease in HDAC activity and mitogen-activated protein kinase phosphatase 1 expression. RNO demonstrated robust anti-inflammatory effects on neutrophils from patients with COPD, reversing their resistance to corticosteroids. The combination of RNO and dexamethasone showed additive/synergistic effects, which were consistent with the reversal of corticosteroid-resistant molecular markers by RNO.

CONCLUSION:

RNO reverses corticosteroid resistance and shows strong anti-inflammatory effects alone or in combination with corticosteroids on neutrophils from patients with COPD.

• Authors: Javier Milara, Javier Lluch, Patricia Almudever, Jose Freire, Qian Xiaozhong, Julio Cortijo
• Journal: The Journal of allergy and clinical immunology (J Allergy Clin Immunol) Vol. 134 Issue 2 Pg. 314-22 (Aug 2014) ISSN: 1097-6825 [Electronic] United States
• PMID: 24636089 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2014 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.

Chemical References

• Adrenal Cortex Hormones
• Aminopyridines
• Benzamides
• Biomarkers
• Complex Mixtures
• Cyclopropanes
• Interleukin-8
• Lipopolysaccharides
• Macrophage Migration-Inhibitory Factors
• Phosphodiesterase 4 Inhibitors
• Dexamethasone
• Phosphatidylinositol 3-Kinases
• Mitogen-Activated Protein Kinase Phosphatases
• Matrix Metalloproteinase 9
• Histone Deacetylases
• Intramolecular Oxidoreductases
• MIF protein, human
• roflumilast N-oxide

Topics

• Adrenal Cortex Hormones (pharmacology)
• Aged
• Aminopyridines (pharmacology)
• Benzamides (pharmacology)
• Biomarkers (metabolism)
• Complex Mixtures (isolation & purification, pharmacology)
• Cyclopropanes (pharmacology)
• Dexamethasone (pharmacology)
• Drug Resistance (drug effects)
• Drug Synergism
• Female
• Gene Expression (drug effects)
• Histone Deacetylases (metabolism)
• Humans
• Interleukin-8 (metabolism)
• Intramolecular Oxidoreductases (metabolism)
• Lipopolysaccharides (pharmacology)
• Macrophage Migration-Inhibitory Factors (metabolism)
• Male
• Matrix Metalloproteinase 9 (metabolism)
• Middle Aged
• Mitogen-Activated Protein Kinase Phosphatases (metabolism)
• Neutrophils (drug effects, metabolism, pathology)
• Phosphatidylinositol 3-Kinases (metabolism)
• Phosphodiesterase 4 Inhibitors (pharmacology)
• Primary Cell Culture
• Pulmonary Disease, Chronic Obstructive (metabolism, pathology)
• Tobacco (chemistry)