In vitro anti-inflammatory effects of AZD8999, a novel bifunctional muscarinic acetylcholine receptor antagonist /β2-adrenoceptor agonist (MABA) compound in neutrophils from COPD patients.

Javier Milara,Montserrat Miralpeix,Esteban Morcillo,Alfredo De Diego,Mònica Aparici,Julio Cortijo,Sonia Contreras,Marta Calbet

doi:10.1371/journal.pone.0210188

Abstract

Recent evidence indicates that AZD8999 (LAS190792), a novel muscarinic acetylcholine receptor antagonist and β2-adrenoceptor agonist (MABA) in development for chronic respiratory diseases, induces potent and sustained relaxant effects in human bronchi by adressing both muscarinic acetylcholine receptors and β2-adrenoceptor. However, the anti-inflammatory effects of the AZD8999 monotherapy or in combination with corticosteroids are unknown. This study investigates the anti-inflammatory effects of AZD8999 in monotherapy and combined with fluticasone propionate in neutrophils from healthy and chronic obstructive pulmonary disease (COPD) patients. Peripheral blood neutrophils from healthy and COPD patients were incubated with AZD8999 and fluticasone propionate, individually or in combination, for 1h followed by lipopolysaccharide (LPS) stimulation for 6h. The IL-8, MMP9, IL-1β, and GM-CSF release was measured in cell culture supernatants. AZD8999 shows ~ 50% maximum inhibitory effect and similar potency inhibiting the released cytokines in neutrophils from healthy and COPD patients. However, while fluticasone propionate suppresses mediator release in neutrophils from healthy patients, COPD neutrophils are less sensitive. The combination of non-effective concentrations of AZD8999 (0.01nM) with non-effective concentrations of fluticasone propionate (0.1nM) shows synergistic anti-inflammatory effects. The studied mechanisms that may be involved in the synergistic anti-inflammatory effects of this combination include the increase of glucocorticoid receptor (GR)α and MKP1 expression, the induction of glucocorticoid response element (GRE) activation and the decrease of ERK1/2, P38 and GR-Ser226 phosphorylations compared with monotherapies. In summary, AZD8999 shows anti-inflammatory effects in neutrophils from COPD patients and induces synergistic anti-inflammatory effects when combined with fluticasone propionate, supporting the use of MABA/ICS combination therapy in COPD.

Highlights

Chronic obstructive pulmonary disease (COPD) is a debilitating disease characterized by persistent airway and systemic inflammation, altering the lung architecture to promote airway obstruction
Triple therapy based on inhaled corticosteroids (ICS) in combination with LABAs and long-acting muscarinic acetylcholine receptor antagonists (LAMAs) is indicated in severe COPD at risk of exacerbations showing benefits in lung function, reduction of exacerbations and an improved quality of life [3]
A recent randomised clinical trial showed that ICS withdrawal did not increase the number of exacerbations in patients with severe COPD under LABA/LAMA/ICS triple therapy [4] which indicate the current controversy in the use of ICS

Summary

Introduction

Chronic obstructive pulmonary disease (COPD) is a debilitating disease characterized by persistent airway and systemic inflammation, altering the lung architecture to promote airway obstruction It is a major cause of morbidity and mortality with a high and increasing prevalence [1]. Triple therapy based on ICS in combination with LABAs and LAMAs is indicated in severe COPD at risk of exacerbations showing benefits in lung function, reduction of exacerbations and an improved quality of life [3]. A recent randomised clinical trial showed that ICS withdrawal did not increase the number of exacerbations in patients with severe COPD under LABA/LAMA/ICS triple therapy [4] which indicate the current controversy in the use of ICS

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