title: Pharmacologic inhibition of S-nitrosoglutathione reductase protects against experimental asthma in BALB/c mice through attenuation of both bronchoconstriction and inflammation.
abstract: 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.
authors.affiliation: N30 Pharmaceuticals, Inc, 3122 Sterling Circle, Suite 200, Boulder, CO 80301, USA. joan.blonder@n30pharma.com.
authors.forename: Joan P
authors.initials: JP
authors.lastname: Blonder
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Mutka
authors.forename: Xicheng
authors.initials: X
authors.lastname: Sun
authors.forename: Jian
authors.initials: J
authors.lastname: Qiu
authors.forename: Lucia H
authors.initials: LH
authors.lastname: Green
authors.forename: Navdeep K
authors.initials: NK
authors.lastname: Mehra
authors.forename: Ramakrishna
authors.initials: R
authors.lastname: Boyanapalli
authors.forename: Michael
authors.initials: M
authors.lastname: Suniga
authors.forename: Kirsten
authors.initials: K
authors.lastname: Look
authors.forename: Chris
authors.initials: C
authors.lastname: Delany
authors.forename: Jane P
authors.initials: JP
authors.lastname: Richards
authors.forename: Doug
authors.initials: D
authors.lastname: Looker
authors.forename: Charles
authors.initials: C
authors.lastname: Scoggin
authors.forename: Gary J
authors.initials: GJ
authors.lastname: Rosenthal
mesh_terms: D000445:Aldehyde Oxidoreductases / Q000037:antagonists & inhibitors*; D000818:Animals; D001249:Asthma / Q000188:drug therapy* / Q000276:immunology / Q000503:physiopathology; D001549:Benzamides / Q000494:pharmacology* / Q000627:therapeutic use*; D016084:Bronchoconstriction / Q000187:drug effects*; D005260:Female; D007249:Inflammation / Q000517:prevention & control*; D051379:Mice; D008807:Mice, Inbred BALB C; D011758:Pyrroles / Q000494:pharmacology* / Q000627:therapeutic use*
chemical_list: D001549:Benzamides; C571360:N6022; D011758:Pyrroles; D000445:Aldehyde Oxidoreductases; C087006:formaldehyde dehydrogenase, glutathione-independent