Pharmacologic inhibition of S-nitrosoglutathione reductase protects against experimental asthma in BALB/c mice through attenuation of both bronchoconstriction and inflammation

BMC Pulmonary Medicine

Gary J Rosenthal¹  Charles Scoggin¹  Doug Looker¹  Jane P Richards¹  Chris Delany¹  Kirsten Look¹  Michael Suniga¹  Ramakrishna Boyanapalli¹  Navdeep K Mehra¹  Lucia H Green¹  Jian Qiu¹  Xicheng Sun¹  Sarah C Mutka¹  Joan P Blonder¹ 

[1] N30 Pharmaceuticals, Inc, 3122 Sterling Circle, Suite 200, Boulder, CO 80301, USA

关键词: NFκB;    N6022;    Nitric oxide;    S-nitrosoglutathione;    S-nitrosoglutathione reductase;    Ovalbumin;    Mouse;    Inflammation;    Asthma;   

Others  :  865456
DOI  :  10.1186/1471-2466-14-3

 received in 2012-11-01, accepted in 2014-01-03,  发布年份 2014

【 摘 要 】

Background

S-nitrosoglutathione (GSNO) serves as a reservoir for nitric oxide (NO) and thus is a key homeostatic regulator of airway smooth muscle tone and inflammation. Decreased levels of GSNO in the lungs of asthmatics have been attributed to increased GSNO catabolism via GSNO reductase (GSNOR) leading to loss of GSNO- and NO- mediated bronchodilatory and anti-inflammatory actions. GSNOR inhibition with the novel small molecule, N6022, was explored as a therapeutic approach in an experimental model of asthma.

Methods

Female BALB/c mice were sensitized and subsequently challenged with ovalbumin (OVA). Efficacy was determined by measuring both airway hyper-responsiveness (AHR) upon methacholine (MCh) challenge using whole body plethysmography and pulmonary eosinophilia by quantifying the numbers of these cells in the bronchoalveolar lavage fluid (BALF). Several other potential biomarkers of GSNOR inhibition were measured including levels of nitrite, cyclic guanosine monophosphate (cGMP), and inflammatory cytokines, as well as DNA binding activity of nuclear factor kappa B (NFκB). The dose response, onset of action, and duration of action of a single intravenous dose of N6022 given from 30 min to 48 h prior to MCh challenge were determined and compared to effects in mice not sensitized to OVA. The direct effect of N6022 on airway smooth muscle tone also was assessed in isolated rat tracheal rings.

Results

N6022 attenuated AHR (ED₅₀ of 0.015 ± 0.002 mg/kg; Mean ± SEM) and eosinophilia. Effects were observed from 30 min to 48 h after treatment and were comparable to those achieved with three inhaled doses of ipratropium plus albuterol used as the positive control. N6022 increased BALF nitrite and plasma cGMP, while restoring BALF and plasma inflammatory markers toward baseline values. N6022 treatment also attenuated the OVA-induced increase in NFκB activation. In rat tracheal rings, N6022 decreased contractile responses to MCh.

Conclusions

The significant bronchodilatory and anti-inflammatory actions of N6022 in the airways are consistent with restoration of GSNO levels through GSNOR inhibition. GSNOR inhibition may offer a therapeutic approach for the treatment of asthma and other inflammatory lung diseases. N6022 is currently being evaluated in clinical trials for the treatment of inflammatory lung disease.

【 授权许可】

   
2014 Blonder et al.; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]Hess DT, Matsumoto A, Kim SO, Marshall HE, Stamler JS: Protein S-nitrosylation: purview and parameters. Nat Rev Mol Cell Biol 2005, 6:150-166.
• [2]Hogg N: The biochemistry and physiology of S-nitrosothiols. Annu Rev Pharmacol Toxicol 2002, 42:585-600.
• [3]Gaston B, Reilly J, Drazen JM, Fackler J, Ramdev P, Arnelle D, Mullins ME, Sugarbaker DJ, Chee C, Singel DJ: Endogenous nitrogen oxides and bronchodilator S-nitrosothiols in human airways. Proc Natl Acad Sci USA 1993, 90:10957-10961.
• [4]Kelm M, Schrader J: Control of coronary vascular tone by nitric oxide. Circ Res 1990, 66:1561-1575.
• [5]Foster MW, Hess DT, Stamler JS: Protein S-nitrosylation in health and disease: a current perspective. Trends Mol Med 2009, 15:391-404.
• [6]Pacher P, Beckman JS, Liaudet L: Nitric oxide and peroxynitrite in health and disease. Physiol Rev 2007, 87:315-424.
• [7]Fortenberry JD, Owens ML, Chen NX, Brown LA: S-nitrosoglutathione inhibits TNF-alpha-induced NFkappaB activation in neutrophils. Inflamm Res 2001, 50:89-95.
• [8]Ricciardolo FL, Sterk PJ, Gaston B, Folkerts G: Nitric oxide in health and disease of the respiratory system. Physiol Rev 2004, 84:731-765.
• [9]Whalen EJ, Foster MW, Matsumoto A, Ozawa K, Violin JD, Que LG, Nelson CD, Benhar M, Keys JR, Rockman HA, Koch WJ, Daaka Y, Lefkowitz RJ, Stamler JS: Regulation of β-adrenergic receptor signaling by s-nitrosylation of g protein-coupled receptor kinase 2. Cell 2007, 129:511-522.
• [10]Jensen DE, Belka GK, Du Bois GC: S-nitrosoglutathione is a substrate for rat alcohol dehydrogenase class III isoenzyme. Biochem J 1998, 331:659-668.
• [11]Liu L, Hausladen A, Zeng M, Que L, Heitman J, Stamler JS: A metabolic enzyme for S-nitrosothiol conserved from bacteria to humans. Nature 2001, 410:490-494.
• [12]Gaston B, Singel D, Doctor A, Stamler JS: S-nitrosothiol signaling in respiratory biology. Am J Respir Crit Care Med 2006, 173:1186-1193.
• [13]Que LG, Yang Z, Stamler JS, Lugogo NL, Kraft M: S-nitrosoglutathione reductase – an important regulator in human asthma. Am J Respir Crit Care Med 2009, 180:226-231.
• [14]Gaston B, Sears S, Woods J, Hunt J, Ponaman M, McMahon T, Stamler JS: Bronchodilator S-nitrosothiol deficiency in asthmatic respiratory failure. Lancet 1998, 351:1317-1319.
• [15]Choudhry S, Que LG, Yang Z, Liu L, Eng C, Kim SO, Kumar G, Thyne S, Chapela R, Rodriguez-Santana JR, Rodriguez-Cintron W, Avila PC, Stamler JS, Burchard EG: GSNO reductase and beta2-adrenergic receptor gene-gene interaction: bronchodilator responsiveness to albuterol. Pharmacogenet Genomics 2010, 6:351-358.
• [16]Moore PE, Ryckman KK, Williams SM, Patel N, Summar ML, Sheller JR: Genetic variants of GSNOR and ADRB2 influence response to albuterol in African-American children with severe asthma. Pediatr Pulmonol 2009, 44:649-654.
• [17]Wu H, Romieu I, Sienra-Monge JJ, Estela DR-N, Anderson DM, Jenchura CA, Li H, Ramirez-Aguilar M, Del CL-SI, London SJ: Genetic variation in S-nitrosoglutathione reductase (GSNOR) and childhood asthma. J Allergy Clin Immunol 2007, 120:322-328.
• [18]Que LG, Liu L, Yan Y, Whitehead GS, Gavett SH, Schwartz DA, Stamler JS: Protection from experimental asthma by an endogenous bronchodilator. Science 2005, 308:1618-1621.
• [19]Gelb AF, Taylor CF, Nussbaum E, Gutierrez C, Schein A, Shinar CM, Schein MJ, Epstein JD, Zamel N: Alveolar and airway sites of nitric oxide inflammation in treated asthma. Am J Respir Crit Care Med 2004, 170:737-741.
• [20]Redington AE: Modulation of nitric oxide pathways: therapeutic potential in asthma and chronic obstructive pulmonary disease. Eur J Pharmacol 2006, 533:263-276.
• [21]Smith AD, Cowan JO, Brassett KP, Herbison GP, Taylor DR: Use of exhaled nitric oxide measurements to guide treatment in chronic asthma. N Engl J Med 2005, 352:2163-2173.
• [22]Maddaford S, Annedi SC, Ramnauth J, Rakhit S: Advances in the development of nitric oxide synthase inhibitors. In Annual Reports in Medicinal Chemistry. Volume 44. Edited by Macor JE. Salt Lake City: Elsevier, Inc; 2009:27-50.
• [23]Vallance P, Leiper J: Blocking NO synthesis: how, where and why? Nat Rev Drug Discov 2002, 1:939-950.
• [24]Al-Sa’doni H, Ferro A: S-Nitrosothiols: a class of nitric oxide-donor drugs. Clin Sci (Lond) 2000, 98:507-520.
• [25]Nevin BJ, Broadley KJ: Nitric oxide in respiratory diseases. Pharmacol Ther 2002, 95:259-293.
• [26]Bredt DS: Endogenous nitric oxide synthesis: biological functions and pathophysiology. Free Radic Res 1999, 31:577-596.
• [27]Lima B, Lam GK, Xie L, Diesen DL, Villamizar N, Nienaber J, Messina E, Bowles D, Kontos CD, Hare JM, Stamler JS, Rockman HA: Endogenous S-nitrosothiols protect against myocardial injury. Proc Natl Acad Sci USA 2009, 106:6297-6302.
• [28]Naseem KM: The role of nitric oxide in cardiovascular diseases. Mol Aspects Med 2005, 26:33-65.
• [29]Savidge TC, Newman P, Pothoulakis C, Ruhl A, Neunlist M, Bourreille A, Hurst R, Sofroniew MV: Enteric glia regulate intestinal barrier function and inflammation via release of S-nitrosoglutathione. Gastroenterology 2007, 132:1344-1358.
• [30]Savidge TC, Sofroniew MV, Neunlist M: Starring roles for astroglia in barrier pathologies of gut and brain. Lab Invest 2007, 87:731-736.
• [31]Green LS, Chun LE, Patton AK, Sun X, Rosenthal GJ, Richards JP: Mechanism of inhibition for N6022, a first-in-class drug targeting S-nitrosoglutathione reductase. Biochemistry 2012, 51:2157-2168.
• [32]Sun X, Wasley J, Qiu J, Blonder J, Stout A, Green L, Strong S, Colagiovanni D, Richards J, Mutka S, Chun L, Rosenthal G: Discovery of S-nitrosoglutathione reductase inhibitors: potential agents for the treatment of asthma and other inflammatory diseases. ACS Med Chem Lett 2011, 2:402-406.
• [33]Hamelmann E, Schwarze J, Takeda K, Oshiba A, Larsen GL, Irvin CG, Gelfand EW: Noninvasive measurement of airway responsiveness in allergic mice using barometric plethysmography. Am J Respir Crit Care Med 1997, 156:766-775.
• [34]Colagiovanni DB, Drolet DW, Langlois-Forget E, Piche MP, Looker D, Rosenthal GJ: A nonclinical safety and pharmacokinetic evaluation of N6022: a first-in-class S-nitrosoglutathione reductase inhibitor for the treatment of asthma. Regul Toxicol Pharmacol 2012, 62:115-124.
• [35]Basu S, Wang X, Gladwin MT, Kim-Shapiro DB: Chemiluminescent detection of S-nitrosated proteins: comparison of tri-iodide, copper/CO/cysteine, and modified copper/cysteine methods. Methods Enzymol 2008, 440:137-156.
• [36]Feil R, Kemp-Harper B: cGMP signalling: from bench to bedside. Conference on cGMP generators, effectors and therapeutic implications. EMBO Rep 2006, 7:149-153.
• [37]Janssen-Heininger YM, Poynter ME, Aesif SW, Pantano C, Ather JL, Reynaert NL, Ckless K, Anathy V, Van DV, Irvin CG, Van DV: Nuclear factor kappaB, airway epithelium, and asthma: avenues for redox control. Proc Am Thorac Soc 2009, 6:249-255.
• [38]Marshall HE, Hess DT, Stamler JS: S-nitrosylation: physiological regulation of NF-kappaB. Proc Natl Acad Sci U S A 2004, 101:8841-8842.
• [39]Lundberg JO, Weitzberg E, Gladwin MT: The nitrate-nitrite-nitric oxide pathway in physiology and therapeutics. Nat Rev Drug Discov 2008, 7:156-167.
• [40]Mattos W, Lim S, Russell R, Jatakanon A, Chung KF, Barnes PJ: Matrix metalloproteinase-9 expression in asthma: effect of asthma severity, allergen challenge, and inhaled corticosteroids. Chest 2002, 122:1543-1552.
• [41]Ying S, Meng Q, Zeibecoglou K, Robinson DS, Macfarlane A, Humbert M, Kay AB: Eosinophil chemotactic chemokines (eotaxin, eotaxin-2, RANTES, monocyte chemoattractant protein-3 (MCP-3), and MCP-4), and C-C chemokine receptor 3 expression in bronchial biopsies from atopic and nonatopic (Intrinsic) asthmatics. J Immunol 1999, 163:6321-6329.
• [42]Kleinbongard P, Dejam A, Lauer T, Rassaf T, Schindler A, Picker O, Scheeren T, Godecke A, Schrader J, Schulz R, Heusch G, Schaub GA, Bryan NS, Feelisch M, Kelm M: Plasma nitrite reflects constitutive nitric oxide synthase activity in mammals. Free Radic Biol Med 2003, 35:790-796.
• [43]Metzger IF, Sertorio JT, Tanus-Santos JE: Relationship between systemic nitric oxide metabolites and cyclic GMP in healthy male volunteers. Acta Physiol (Oxf) 2006, 188:123-127.
• [44]Kelleher ZT, Matsumoto A, Stamler JS, Marshall HE: NOS2 regulation of NF-kappaB by S-nitrosylation of p65. J Biol Chem 2007, 282:30667-30672.
• [45]Sanghani PC, Davis WI, Fears SL, Green SL, Zhai L, Tang Y, Martin E, Bryan NS, Sanghani SP: Kinetic and cellular characterization of novel inhibitors of S-nitrosoglutathione reductase. J Biol Chem 2009, 284:24354-24362.
• [46]Chen Q, Sievers RE, Varga M, Kharait S, Haddad DJ, Patton AK, Delany CS, Mutka SC, Blonder JP, Dube GP, Rosenthal GJ, Springer ML: Pharmacological inhibition of S-nitrosoglutathione reductase improves endothelial vasodilatory function in rats in vivo. J Appl Physiol 2013, 114:752-760.
• [47]Yates DH: Role of exhaled nitric oxide in asthma. Immunol Cell Biol 2001, 79:178-190.
• [48]Ricciardolo FL, Di SA, Sabatini F, Folkerts G: Reactive nitrogen species in the respiratory tract. Eur J Pharmacol 2006, 533:240-252.
• [49]Foster MW, Yang Z, Gooden DM, Thompson JW, Ball CH, Turner ME, Hou Y, Pi J, Moseley MA, Que L: Proteonomic characterization by S-nitrosogluthatione reductase inhibition. J Proteome Res 2012, 11:2480-2491.
• [50]Foster MW, Yang Z, Potts EN, Foster WM, Que LG: S-nitrosoglutathione supplementation to ovalbumin-sensitized and -challenged mice ameliorates methacholine-induced bronchoconstriction. Am J Physiol Lung Cell Mol Physiol 2011, 301:L739-L744.
• [51]Blonder JP, Mutka SC, Drolet DW, Damaj B, Spicer D, Russell V, Sun X, Rosenthal GJ, Scoggin C: Oral S-nitrosoglutathione reductase inhibitors attenuate pulmonary inflammation and decrease airspace enlargement in experimental models of chornic obstructive pulmonary disease. Am J Respir Crit Care Med 2011, 183(1):A6433-A6433.
• [52]Blonder JP, Mutka SC, Sun X, Drolet DW, Colagiovanni DB, Lyng G, Scoggin C, Rosenthal GJ: S-nitrosoglutathione reductase inhibitors for the prevention and treatment of experimental DSS colitis. In Advances in Inflammatory Bowel Diseases Crohn’s & Colitis Foundations Clinical & Research Conference. Imedex, LLC: Proceedings; 2011:P251-P251.
• [53]Cox AG, Saunders DC, Kelsey PB, Conway AA, Tesmenitsky Y, Marchini JF, Brown KK, Stamler JS, Colagiovanni DB, Rosenthal GJ, Croce KJ, North TE, Goessling W: S-nitrosothiol signaling regulates liver development and improves outcome following toxic liver injury. Cell Reports 2013, 6:1-14.
• [54]Bates J, Irvin C, Brusasco V, Drazen J, Fredberg J, Loring S, Eidelman D, Ludwig M, Macklem P, Martin J, Milic-Emili J, Hantos Z, Hyatt R, Lai-Fook S, Leff A, Solway J, Lutchen K, Suki B, Mitzner W, Pare P, Pride N, Sly P: The use and misuse of Penh in animal models of lung disease. Am J Respir Cell Mol Biol 2004, 31:373-374.
• [55]Bates JH, Rincon M, Irvin CG: Animal models of asthma. Am J Physiol Lung Cell Mol Physiol 2009, 297:L401-L410.
• [56]Pinder AG, Rogers SC, Khalatbari A, Ingram TE, James PE: The measurement of nitric oxide and its metabolites in biological samples by ozone-based chemiluminescence. Methods Mol Biol 2008, 476:11-28.
• [57]Yang BK, Vivas EX, Reiter CD, Gladwin MT: Methodologies for the sensitive and specific measurement of S-nitrosothiols, iron-nitrosyls, and nitrite in biological samples. Free Radic Res 2003, 37:1-10.
• [58]Broniowska KA, Diers AR, Hogg N: S-nitrosoglutathione. Biochim Biophys Acta 1830, 2013:3173-3181.
• [59]Henderson EM, Gaston B: SNOR and wheeze: the asthma enzyme? Trends Mol Med 2005, 11:481-484.
• [60]Dweik RA, Comhair SA, Gaston B, Thunnissen FB, Farver C, Thomassen MJ, Kavuru M, Hammel J, Abu-Soud HM, Erzurum SC: NO chemical events in the human airway during the immediate and late antigen-induced asthmatic response. Proc Natl Acad Sci USA 2001, 98:2622-2627.