RationaleDenufosol stimulates chloride secretion independent of the chloride channel which is dysfunctional in cystic fibrosis (CF) and therefore has the potential to benefit CF patients regardless of genotype.ObjectivesTo assess the efficacy of denufosol in CF patients with mild lung function impairment age 5 years and older.MethodsThis multicenter, randomized, parallel group double-blind placebo-controlled trial was conducted at 102 CF care centers in Australia, Canada and the United States (NCT00625612) The active group (n=233) received 60 mg denufosol via inhalation three times daily The primary efficacy endpoint was change in FEV1 in liters from Day 0 to week 48.Measurements and main results685 patients were screened for the study and 466 patients (233 in each group) were randomized to study treatment. The adjusted mean change in FEV1was 40 mL for denufosol and 32 mL for placebo with a resulting treatment effect of 8 mL (95% CI −0.040, 0.056). The average rate of change in FEV1 percent of predicted over 0 to 48 weeks was −3.04% for placebo vs. −2.30 for denufosol (a difference of 24% relative to placebo) among all patients. The incidence of pulmonary exacerbation was 26% vs. 21% for the placebo and denufosol groups with no differences in the time to first event. The study treatments were well tolerated and there was no evidence of systemic effects in any safety parameter assessed.ConclusionsIn patients with CF treatment with denufosol for 48 weeks did not improve pulmonary function or reduce the incidence of pulmonary exacerbations.

RATIONALEIntervention for cystic fibrosis lung disease early in its course has the potential to delay or prevent progressive changes that lead to irreversible airflow obstruction. Denufosol is a novel ion channel regulator designed to correct the ion transport defect and increase the overall mucociliary clearance in cystic fibrosis lung disease by increasing chloride secretion, inhibiting sodium absorption, and increasing ciliary beat frequency in the airway epithelium independently of cystic fibrosis transmembrane conductance regulator genotype.OBJECTIVESTo evaluate the efficacy and safety of denufosol in patients with cystic fibrosis who had normal to mildly impaired lung function characteristic of early cystic fibrosis.METHODSA total of 352 patients greater than or equal to 5 years old with cystic fibrosis who had FEV(1) greater than or equal to 75% of predicted normal were randomized to receive inhaled denufosol, 60 mg, or placebo three times daily in a Phase 3, randomized, double-blind, placebo-controlled, 24-week trial.MEASUREMENTS AND MAIN RESULTSMain outcome measures included change in FEV(1) from baseline to Week 24 endpoint and adverse events. Mean change from baseline to Week 24 endpoint in FEV(1) (primary efficacy endpoint) was 0.048 L for denufosol (n = 178) and 0.003 L for placebo (n = 174; P = 0.047). No significant differences between groups were observed for secondary endpoints including exacerbation rate and other measures of lung function. Denufosol was well tolerated with adverse event and growth profiles similar to placebo.CONCLUSIONSDenufosol improved lung function relative to placebo in cystic fibrosis patients with normal to mildly impaired lung function. Clinical trial registered with www.clinicaltrials.gov (NCT00357279).

CF lung disease arises from ion transport defect. Disease origins in small airways before 1 yr of age necessitate early intervention. Denufosol (DEN), a novel chloride channel activator, stimulates Cl − transport, inhibits Na + absorption and increases ciliary beat frequency. A Phase 3, double‐blind, placebo (PL)‐controlled trial (TIGER‐1) was conducted in 352 CF pts to compare inhaled DEN 60 mg TID to PL for 24 wks. Pts were ≥ 5 yrs (mean 14.6 yrs) and had FEV 1 ≥ 75% predicted (mean 92%). Pulmonary function tests and exacerbations, QOL measures and systemic exposure of DEN were evaluated. This work examines endpoints relevant to DEN use as an early intervention agent. The ability to reach small airways was confirmed by significant improvement in FEF 25%–75% in pts ≤ 110% predicted FEV 1 (p=0.025). There was little to no systemic exposure to denufosol, no accumulation with chronic dosing, and no evidence of systemic AEs including liver enzyme levels, blood biochemistry and differential cell counts. DEN's mechanism of action addressing the basic defect, the lack of systemic exposure and ability to target small airways makes this a potentially promising therapy for early intervention in CF lung disease.

Among the most promising of the new therapies being developed for the treatment of Cystic Fibrosis (CF) are those targeted at increasing mucosal hydration on the surface of the airways. One of these therapies, P2Y(2) receptor agonists, bypasses the defective CFTR chloride channel, and activates an alternative chloride channel. This activation results in an increase in airway surface epithelial hydration, and through these actions and effects on cilia beat frequency, increases mucociliary clearance. The pharmacology of P2Y(2) agonists has been confirmed in several preclinical and clinical studies. Denufosol tetrasodium is a novel second-generation, metabolically stable, selective P2Y(2) receptor agonist currently in Phase 3 clinical development. In radiolabelled deposition studies of P2Y(2) agonists in healthy non-smokers and smokers, approximately 7mg of a 40-mg nebulizer (PARI LC Star) load was deposited in the lungs. In a pharmacokinetic study in healthy volunteers, very limited systemic exposure was observed when doses of 200mg of denufosol were nebulized. Thus, it appears that high concentrations of denufosol can be achieved in the airways with very low systemic absorption. Denufosol has been generally well-tolerated in healthy volunteers and patients with CF. The most common adverse events were in the respiratory system, with cough having the highest frequency. Doses of 20-60mg have been evaluated in Phase 2 trials of up to 28 days duration, and superiority relative to placebo on FEV1 has been observed in patients with relatively normal lung function (FEV1 greater than or equal to 75% of predicted). The first Phase 3 trial is a comparison of denufosol 60mg and placebo in 350 patients with CF with FEV1 at study entry greater than or equal to 75% of predicted.

Aims: fi~stic Fibrosis (CF) is a common autosomal recessive disorder in North America and Europe.Defective CFTR function in CF airway epithelia can be corrected in vitro by inlroducing wild4ype CFTR gene.Recombinant adeno associated virus vector encoding the complete human CFTR eDNA (tgAAVCF) has been safe and well tolerated ha studies of delivery to the nose, sinus and lungs of CF patients.A small (n 37) randomized, double blind placebo controlled Phase II trial of 3 monthly doses aerosolized tgAAVCF showed positive trends ha pulmonary function and induced sputum IL 8 levels (Chest 2(KH;125:509).A larger study was thus designed.Methods: CF subjects (n 102) with FEV 1 >60% were randomized 1:1 to receive 2 aerosolized doses of 101~ DNAse resistant particles tgAAVCF or placebo given 30 days apart.The primary endpoint is 30 day change ha FEV 1. Secondary endpoints are change in pulmonary function over time, sputum biomar ker s, days of antibiotic use and safety.Safety is reviewed by an independent Data Monitoring Committee (DMC) every four months.Results: The DMC has not raised any safety concerns and the trial enrolled to target goal (n>l(D).Preliminary AE data col~ir m tgAAVCF is safe and well tolerated.Conclusion: Active and placebo group results will be presented.

The aim of the present study was to investigate the possible interaction between intracellular Ca2+ and nitric oxide (NO) in rat pancreatic acinar cells, especially intracellular signaling events. (1) Nitric oxide donors SNP (0.1–100 μM) and NOR-3 (50–400 μM) induced Ca2+ oscillations in fluo-4-loaded acini, that appeared to be analogous to what we usually observe in acini stimulated with physiological secretagogues such as CCK-8 and this oscillations were abolished in the presence of carboxy-PTIO. (2) The NO donors-evoked Ca2+ oscillations were not abolished even in the absence of extracellular Ca2+ but totally disappeared when cells were pretreated with thapsigargin, a sarcoplasmic-endoplasmic reticulum Ca2+ ATPase (SERCA) inhibitor. (3) Inhibition of guanylate cyclase with 1 H-[1,2,4] oxadiazolo [4,3-a] quinoxaline-1-one (ODQ) attenuated Ca2+ oscillations evoked by SNP in the absence of extracellular Ca2+. (4) Inhibitors of phospholipase C activity, U73122 and the IP3R blocker xestospongin C, both abolished the SNP-induced Ca2+ response. (5) Furthermore, we found that both CCK-8 and carbachol (CCh) induced NO production in DAF-2-loaded acinar cells and that an inhibitor of NO synthase, NG-monomethyl-l-arginine (L-NMMA), significantly reduced CCK-8-induced Ca2+ oscillation. These results indicate that NO mobilizes Ca2+ from internal stores through activation of guanylate cyclase and resultant cGMP production. In addition, PLC activation of IP3 production is also suggested to be involved in Ca2+ mobilization via IP3 receptors. This suggests the presence of cross-talk between Ca2+ and NO in pancreatic acini and this cascade may, at least partially, participate in physiological secretagogue-evoked Ca2+ dynamics in pancreatic acinar cells.

Aims: fi~stic Fibrosis (CF) is a common autosomal recessive disorder in North America and Europe.Defective CFTR function in CF airway epithelia can be corrected in vitro by inlroducing wild4ype CFTR gene.Recombinant adeno associated virus vector encoding the complete human CFTR eDNA (tgAAVCF) has been safe and well tolerated ha studies of delivery to the nose, sinus and lungs of CF patients.A small (n 37) randomized, double blind placebo controlled Phase II trial of 3 monthly doses aerosolized tgAAVCF showed positive trends ha pulmonary function and induced sputum IL 8 levels (Chest 2(KH;125:509).A larger study was thus designed.Methods: CF subjects (n 102) with FEV 1 >60% were randomized 1:1 to receive 2 aerosolized doses of 101~ DNAse resistant particles tgAAVCF or placebo given 30 days apart.The primary endpoint is 30 day change ha FEV 1. Secondary endpoints are change in pulmonary function over time, sputum biomar ker s, days of antibiotic use and safety.Safety is reviewed by an independent Data Monitoring Committee (DMC) every four months.Results: The DMC has not raised any safety concerns and the trial enrolled to target goal (n>l(D).Preliminary AE data col~ir m tgAAVCF is safe and well tolerated.Conclusion: Active and placebo group results will be presented.

This study was undertaken to evaluate the efficacy and safety of fluticasone propionate, an inhaled corticosteroid, in adolescents and adults with moderate asthma who were previously taking inhaled corticosteroids. After a 2-week, open-label screening period, a double-masked, randomized, parallel-group, dose-ranging study was conducted over 12 weeks in 21 outpatient centers throughout the United States. Patients (N = 304) ≥12 years of age with moderate asthma previously treated with inhaled corticosteroids and beta-sympathomimetic bronchodilators were enrolled. Patients were assigned to receive placebo or fluticasone propionate 100, 250, or 500 μg twice daily via a metered-dose inhaler without a spacer device. These doses refer to the amount of fluticasone propionate released from the valve of the metered-dose inhaler; the corresponding doses released from the activator of the metered-dose inhaler are 88 μg, 220 μg, and 440 μg, respectively. Between baseline and end point, mean values of forced expiratory volume in 1 second decreased 0.31 L in the placebo group and improved 0.39 L, 0.30 L, and 0.43 L in patients receiving 100-μg, 250-μg, and 500-μg, fluticasone propionate, respectively. The differences between placebo and all treatment groups were statistically significant. More patients were withdrawn from placebo (72%) than from fluticasone propionate (13% to 16%) because of failure to meet predetermined asthma stability criteria. Differences in baseline-to-end point changes in morning peak expiratory flow rate, physician overall assessments and patient-rated assessment of symptoms, and albuterol use for symptoms control also significantly favored each fluticasone propionate group over placebo. There were essentially no differences in efficacy among the three fluticasone propionate groups. Treatment-related adverse events occurred in 8% of placebo-treated patients and 13% to 15% of fluticasone propionate—treated patients; these events were mainly localized to the oropharynx/larynx. A 12-week course of fluticasone propionate (100, 250, and 500 μg twice daily) was well tolerated and more effective than placebo based on maintenance of asthma stability, pulmonary function tests, physician and patient assessments, and rescue bronchodilator use. No dose-related effects were observed with the dosages of fluticasone propionate used in this study.