Roflumilast attenuates allergen-induced inflammation in mild asthmatic subjects

Gail M Gauvreau, Louis-Philippe Boulet, Christine Schmid-Wirlitsch, Johanne Côté, Mylinh Duong, Kieran J Killian, Joanne Milot, Francine Deschesnes, Tara Strinich, Richard M Watson, Dirk Bredenbröker, Paul M O'Byrne, Gail M Gauvreau, Louis-Philippe Boulet, Christine Schmid-Wirlitsch, Johanne Côté, Mylinh Duong, Kieran J Killian, Joanne Milot, Francine Deschesnes, Tara Strinich, Richard M Watson, Dirk Bredenbröker, Paul M O'Byrne

Abstract

Background: Phosphodiesterase 4 (PDE4) inhibitors increase intracellular cyclic adenosine monophosphate (cAMP), leading to regulation of inflammatory cell functions. Roflumilast is a potent and targeted PDE4 inhibitor. The objective of this study was to evaluate the effects of roflumilast on bronchoconstriction, airway hyperresponsiveness (AHR), and airway inflammation in mild asthmatic patients undergoing allergen inhalation challenge.

Methods: 25 subjects with mild allergic asthma were randomized to oral roflumilast 500 mcg or placebo, once daily for 14 days in a double-blind, placebo-controlled, crossover study. Allergen challenge was performed on Day 14, and FEV1 was measured until 7 h post challenge. Methacholine challenge was performed on Days 1 (pre-dose), 13 (24 h pre-allergen), and 15 (24 h post-allergen), and sputum induction was performed on Days 1, 13, 14 (7 h post-allergen), and 15.

Results: Roflumilast inhibited the allergen-induced late phase response compared to placebo; maximum % fall in FEV1 (p = 0.02) and the area under the curve (p = 0.01). Roflumilast had a more impressive effect inhibiting allergen-induced sputum eosinophils, neutrophils, and eosinophil cationic protein (ECP) at 7 h post-allergen (all p = 0.02), and sputum neutrophils (p = 0.04), ECP (p = 0.02), neutrophil elastase (p = 0.0001) and AHR (p = 0.004) at 24 h post-allergen.

Conclusions: This study demonstrates a protective effect of roflumilast on allergen-induced airway inflammation. The observed attenuation of sputum eosinophils and neutrophils demonstrates the anti-inflammatory properties of PDE4 inhibition and supports the roles of both cell types in the development of late phase bronchoconstriction and AHR.

Trial registration: ClinicalTrials.gov: NCT01365533.

Figures

Figure 1
Figure 1
Study schematic and subject disposition. D: day; PLA: placebo; ROF: roflumilast.
Figure 2
Figure 2
Mean sputum inflammatory cells (± SEM) at baseline before treatment with roflumilast (solid bars) or placebo (open bars), after 13 days' treatment, and 7 h and 24 h after allergen challenge. Roflumilast significantly inhibited the allergen-induced influx of sputum eosinophils, neutrophils and metachromatic cells (p < 0.05).
Figure 3
Figure 3
Mean sputum fluid phase mediators (± SEM) at baseline before treatment with roflumilast (solid bars) or placebo (open bars), after 13 days' treatment, and 7 h and 24 h after allergen challenge. Roflumilast significantly inhibited the allergen-induced increased level of eosinophil cationic protein (ECP) and neutrophil elastase (NE) (p < 0.05).
Figure 4
Figure 4
Mean percent fall in FEV1 (± SEM) during the early and late asthmatic responses. Roflumilast significantly inhibited the area of the early and late responses (p < 0.05). FEV1: forced expiratory volume in 1 second.
Figure 5
Figure 5
Mean plasma concentration (± SEM) of roflumilast and the active metabolite roflumilast N-oxide at Day 14.

References

    1. Fahy JV, Wong H, Liu J, Boushey HA. Comparison of samples collected by sputum induction and bronchoscopy from asthmatic and healthy subjects. Am J Respir Crit Care Med. 1995;152:53–8.
    1. Shannon J, Ernst P, Yamauchi Y, Olivenstein R, Lemiere C, Foley S, Cicora L, Ludwig M, Hamid Q, Martin JG. Differences in airway cytokine profile in severe asthma compared to moderate asthma. Chest. 2008;133:420–426. doi: 10.1378/chest.07-1881.
    1. Qiu Y, Zhu J, Bandi V, Guntupalli KK, Jeffery PK. Bronchial mucosal inflammation and upregulation of CXC chemoattractants and receptors in severe exacerbations of asthma. Thorax. 2007;62:475–482. doi: 10.1136/thx.2006.066670.
    1. Keatings VM, Jatakanon A, Worsdell YM, Barnes PJ. Effects of inhaled and oral glucocorticoids on inflammatory indices in asthma and COPD. Am J Respir Crit Care Med. 1997;155:542–548.
    1. Nguyen LT, Lim S, Oates T, Chung KF. Increase in airway neutrophils after oral but not inhaled corticosteroid therapy in mild asthma. Respir Med. 2005;99:200–207. doi: 10.1016/j.rmed.2004.06.007.
    1. Schudt C, Gantner F, Tenors H, Hatzelmann A. Therapeutic potential of selective PDE inhibitors in asthma. Pulm Pharmacol Ther. 1999;12:123–129. doi: 10.1006/pupt.1999.0182.
    1. Beavo JA. Cyclic nucleotide phosphodiesterases: functional implications of multiple isoforms. Physiol Rev. 1995;75:725–748.
    1. Calverley PM, Rabe KF, Goehring UM, Kristiansen S, Fabbri LM, Martinez FJ. M2-124 and M2-125 study groups. Roflumilast in symptomatic chronic obstructive pulmonary disease: two randomised clinical trials. Lancet. 2009;374:685–694. doi: 10.1016/S0140-6736(09)61255-1.
    1. Fabbri LM, Calverley PM, Izquierdo-Alonso JL, Bundschuh DS, Brose M, Martinez FJ, Rabe KF. M2-127 and M2-128 study groups. Roflumilast in moderate-to-severe chronic obstructive pulmonary disease treated with longacting bronchodilators: two randomised clinical trials. Lancet. 2009;374:695–703. doi: 10.1016/S0140-6736(09)61252-6.
    1. Pryzwansky KB, Madden VJ. Type 4A cAMP-specific phosphodiesterase is stored in granules of human neutrophils and eosinophils. Cell Tissue Res. 2003;312:301–311. doi: 10.1007/s00441-003-0728-y.
    1. Peter D, Jin SL, Conti M, Hatzelmann A, Zitt C. Differential expression and function of phosphodiesterase 4 (PDE4) subtypes in human primary CD4+ T cells: predominant role of PDE4. J Immunol. 2007;178:4820–4831.
    1. Sawai T, Ikai K, Uehara M. Cyclic adenosine monophosphate phosphodiesterase activity in peripheral blood mononuclear leucocytes from patients with atopic dermatitis: correlation with respiratory atopy. Br J Dermatol. 1998;138:846–848. doi: 10.1046/j.1365-2133.1998.02223.x.
    1. Hanifin JM, Chan SC. Monocyte phosphodiesterase abnormalities and dysregulation of lymphocyte function in atopic dermatitis. J Invest Dermatol. 1995;105(1 Suppl):84S–8S.
    1. Hatzelmann A, Morcillo EJ, Lungarella G, Adnot S, Sanjar S, Beume R, Schudt C, Tenor H. The preclinical pharmacology of roflumilast--a selective, oral phosphodiesterase 4 inhibitor in development for chronic obstructive pulmonary disease. Pulm Pharmacol Ther. 2010;23:235–256. doi: 10.1016/j.pupt.2010.03.011.
    1. Boulet LP, Gauvreau G, Boulay ME, O'Byrne P, Cockcroft DW. The allergen bronchoprovocation model: an important tool for the investigation of new asthma anti-inflammatory therapies. Allergy. 2007;62:1101–1110. doi: 10.1111/j.1398-9995.2007.01499.x.
    1. O'Byrne PM, Dolovich J, Hargreave FE. Late asthmatic responses. Am Rev Respir Dis. 1987;136:740–751. doi: 10.1164/ajrccm/136.3.740.
    1. Gauvreau GM, Lee JM, Watson RM, Irani AM, Schwartz LB, O'Byrne PM. Increased numbers of both airway basophils and mast cells in sputum after allergen inhalation challenge of atopic asthmatics. Am J Respir Crit Care Med. 2000;161:1473–1478.
    1. Gauvreau GM, Watson RM, O'Byrne PM. Kinetics of allergen-induced airway eosinophilic cytokine production and airway inflammation. Am J Respir Crit Care Med. 1999;160:640–647.
    1. Aalbers R, Kauffman HF, Vrugt B, Koeter GH, de Monchy JG. Allergen-induced recruitment of inflammatory cells in lavage 3 and 24 h after challenge in allergic asthmatic lungs. Chest. 1993;103:1178–1184. doi: 10.1378/chest.103.4.1178.
    1. Woolley KL, Adelroth E, Woolley MJ, Ellis R, Jordana M, O'Byrne PM. Effects of allergen challenge on eosinophils, eosinophil cationic protein, and granulocyte-macrophage colony-stimulating factor in mild asthma. Am J Respir Crit Care Med. 1995;151:1915–1924.
    1. Bentley AM, Meng Q, Robinson DS, Hamid Q, Kay AB, Durham SR. Increases in activated T lymphocytes, eosinophils, and cytokine mRNA expression for interleukin-5 and granulocyte/macrophage colony-stimulating factor in bronchial biopsies after allergen inhalation challenge in atopic asthmatics. Am J Respir Cell Mol Biol. 1993;8:35–42.
    1. Nouri-Aria KT, Irani AM, Jacobson MR, O'brien F, Varga EM, Till SJ, Durham SR, Schwartz LB. Basophil recruitment and IL-4 production during human allergen-induced late asthma. J Allergy Clin Immunol. 2001;108:205–211. doi: 10.1067/mai.2001.117175.
    1. Gauvreau GM, Boulet LP, Cote J, Deschesnes F, Duong M, Killian K, Obminski G, Strinich TX, Watson RM, Cerasoli F, Schmid-Wirlitsch C, Bredenbröker D, O'Byrne PM. Effects of roflumilast on airway inflammation and function following allergen challenge in patients with allergic asthma [abstract] Am J Respir Crit Care Med. 2007;175:A484.
    1. Bateman ED, Izquierdo JL, Harnest U, Hofbauer P, Magyar P, Schmid-Wirlitsch C, Leichtl S, Bredenbröker D. Efficacy and safety of roflumilast in the treatment of asthma. Ann Allergy Asthma Immunol. 2006;96:679–686. doi: 10.1016/S1081-1206(10)61065-4.
    1. Cockcroft DW. In: Airway responsiveness: measurement and interpretation. Hargreave FE, Woolcock AJ, editor. Mississauga: Astra Pharmaceuticals Canada Ltd; 1985. Measure of airway responsiveness to inhaled histamine or methacholine; method of continuous aerosol generation and tidal breathing inhalation; pp. 22–28.
    1. Cockcroft DW, Murdock KY, Kirby J, Hargreave F. Prediction of airway responsiveness to allergen from skin sensitivity to allergen and airway responsiveness to histamine. Am Rev Respir Dis. 1987;135:264–267.
    1. Pizzichini E, Pizzichini MM, Efthimiadis A, Evans S, Morris MM, Squillace D, Gleich GJ, Dolovich J, Hargreave FE. Indices of airway inflammation in induced sputum: reproducibility and validity of cell and fluid-phase measurements. Am J Respir Crit Care Med. 1996;154:308–317.
    1. Bethke TD, Giessmann T, Westphal K, Weinbrenner A, Hauns B, Hauschke D, David M, Lahu G, Zech K, Hermann R, Siegmund W. Roflumilast, a once-daily oral phosphodiesterase 4 inhibitor, lacks relevant pharmacokinetic interactions with inhaled salbutamol when co-administered in healthy subjects. Int J Clin Pharmacol Ther. 2006;44:572–579.
    1. Harbinson PL, MacLeod D, Hawksworth R, O'Toole S, Sullivan PJ, Heath P, Kilfeather S, Page CP, Costello J, Holgate ST, Lee TH. The effect of a novel orally active selective PDE4 isoenzyme inhibitor (CDP840) on allergen-induced responses in asthmatic subjects. Eur Respir J. 1997;10:1008–1014. doi: 10.1183/09031936.97.10051008.
    1. Louw C, Williams Z, Venter L, Leichtl S, Schmid-Wirlitsch C, Bredenbroker D, Bardin PG. Roflumilast, a phosphodiesterase 4 inhibitor, reduces airway hyperresponsiveness after allergen challenge. Respiration. 2007;74:411–417. doi: 10.1159/000095677.
    1. van Schalkwyk E, Strydom K, Williams Z, Venter L, Leichtl S, Schmid-Wirlitsch C, Bredenbröker D, Bardin PG. Roflumilast, an oral, once-daily phosphodiesterase 4 inhibitor, attenuates allergen-induced asthmatic reactions. J Allergy Clin Immunol. 2005;116:292–298. doi: 10.1016/j.jaci.2005.04.023.
    1. Grootendorst DC, Gauw SA, Verhoosel RM, Sterk PJ, Hospers JJ, Bredenbroker D, Bethke TD, Hiemstra PS, Rabe KF. Reduction in sputum neutrophil and eosinophil numbers by the PDE4 inhibitor roflumilast in patients with COPD. Thorax. 2007;62:1081–1087. doi: 10.1136/thx.2006.075937.
    1. Sanz MJ, Cortijo J, Taha MA, Cerda-Nicolas M, Schatton E, Burgbacher B, Klar J, Tenor H, Schudt C, Issekutz AC, Hatzelmann A, Morcillo EJ. Roflumilast inhibits leukocyte-endothelial cell interactions, expression of adhesion molecules and microvascular permeability. Br J Pharmacol. 2007;152:481–492. doi: 10.1038/sj.bjp.0707428.
    1. Hatzelmann A, Schudt C. Anti-inflammatory and immunomodulatory potential of the novel PDE4 inhibitor roflumilast in vitro. J Pharmacol Exp Ther. 2001;297:267–279.
    1. Gauvreau GM, Doctor J, Watron R, Jordana M, O'Byrne PM. Effects of inhaled budesonide on allergen-induced airway responses and airway inflammation. Am J Respir Crit Care Med. 1996;154:1267–1271.
    1. Cowan DC, Cowan JO, Palmay R, Williamson A, Taylor DR. Effects of steroid therapy on inflammatory cell subtypes in asthma. Thorax. 2010;65:384–390. doi: 10.1136/thx.2009.126722.
    1. Grootendorst DC, Gauw SA, Baan R, Kelly J, Murdoch RD, Sterk PJ, Rabe KF. Does a single dose of the phosphodiesterase 4 inhibitor, cilomilast (15 mg), induce bronchodilation in patients with chronic obstructive pulmonary disease? Pulm Pharmacol Ther. 2003;16:115–120. doi: 10.1016/S1094-5539(02)00172-4.

Source: PubMed

Спонсоры и соавторы

Заболевания

Медикаментозные вмешательства

Подписаться