Efficacy and safety of a 12-week therapy with a new formulation of fluticasone propionate at doses of 125 and 250 μg administered through a new generation cyclohaler twice daily, in comparison to fluticasone propionate 500 μg dry powder inhaler twice dail

WSTĘP: Flutykazon w postaci wziewnej stosowany jest w leczeniu przewlekłej astmy oskrzelowej. Ma udowodnioną w badaniach klinicznych i obserwacyjnych wysoką skuteczność i dobry profil bezpieczeństwa. Wyróżnia się spośród innych leków należących do tej grupy unikatowymi właściwościami farmakokinetycznymi. Badania in vitro na sztucznym modelu dróg oddechowych jak i badania farmakokinetyczne przeprowadzane na zdrowych ochotnikach wykazały, że w nowej formulacji lek ten cechuje się dwukrotnie lepszą depozycją płucną w porównaniu z preparatem referencyjnym. Celem tego badania była ocena skuteczności i bezpieczeństwa nowej formulacji propionianu flutykazonu podawanego z inhalatora nowej generacji typu cyklohaler (CNG) w porównaniu z oryginalnym produktem flutykazonu w inhalatorze proszkowym DPI u chorych na przewlekłą umiarkowaną astmę.MATERIAŁ I METODY: Do badania włączono 457 pacjentów, 376 osób zrandomizowano przydzielając losowo do jednej z trzech badanych grup: 127 osób do grupy leczonej flutykazonem w nowej formulacji w dawce 125 μg BID, 125 osób do grupy leczonej flutykazonem w nowej formulacji 250 μg BID i 124 osoby do grupy z lekiem referencyjnym — flutykazon DPI 500 μg BID. Badane grupy nie różniły się na początku badania pod względem demograficznym ani klinicznym. Aktywna terapia trwała 12 tygodni. Pierwszorzędowym punktem końcowym była średnia zmiana porannego PEF podczas 12-tygodniowego okresu leczenia (za znamienną statystycznie różnicę uznano ΔmPEF o 15 l/min). Badano ponadto inne parametry czynnościowe układu oddechowego, objawy kliniczne i zużycie leków ratunkowych. W czasie trwania całego badania monitorowano bezpieczeństwo pacjentów rejestrując zdarzenia niepożądane, dodatkowo oceniano ekspozycję systemową na flutykazon, badając w 45-osobowej podgrupie zmiany kortyzolu w surowicy i dobowej zbiórce moczu. Analizę statystyczną przeprowadzono zarówno na grupie intention-to-treat jak i per protocol.WYNIKI: Zarówno w analizie PP, jak i ITT średnia zmiana porannego PEF na koniec okresu leczenia w porównaniu z okresem wstępnym była istotna statystycznie we wszystkich grupach terapeutycznych. Efektywność terapii flutykazonem zarówno w dawce 125 μg BID, jak i 250 μg BID oraz preparatem referencyjnym nie różniła się istotnie statystycznie po 12 tygodni terapii, jak również w całym okresie leczenia. Również w zakresie innych parametrów czynnościowych jak wieczorny PEF, FEV1, objawy kliniczne i zużycie leków ratunkowych obserwowano istotną poprawę we wszystkich grupach terapeutycznych w czasie trwania badania, bez istotnej różnicy w efektywności pomiędzy badanymi grupami. Porównując efektywność dawki flutykazonu 125 μg BID z 250 μg BID produktu odtwórczego wykazano słabą zależność dawka–odpowiedź w zakresie zmiany parametru porannego PEF, co wynika z niemal płaskiej krzywej dawka–odpowiedź w zakresie dawek średnich i wysokich dla tego leku. Nie wykazano też istotnych różnic ilościowych i jakościowych między grupami w rejestrowanych zdarzeniach niepożądanych zakwalifikowanych jako związane z leczeniem flutykazonem. W żadnej z grup nie odnotowano istotnych zmian w stężeniu kortyzolu w surowicy i dobowej zbiórce moczu między poziomem wyjściowym i wizytą końcową.WNIOSKI: Flutykazon CNG w porównaniu z oryginalnym produktem flutykazonu DPI, pozwala na dwukrotne obniżenie dawki tego leku przy zachowaniu efektywności klinicznej odpowiadającej lekowi referencyjnemu w dwukrotnie wyższej dawce. Flutykazon w nowej formulacji podawany z inhalatora typu CNG ma porównywalny klinicznie profil bezpieczeństwa do leku referencyjnego.INTRODUCTION: Inhaled fluticasone is used in the treatment of chronic bronchial asthma. Its high efficacy and good safety profile have been proven by clinical trials and observations. Its unique pharmacokinetic properties make it distinguishable from other drugs from this group. In vitro tests run on an artificial model of the airways and pharmacokinetic studies conducted on healthy volunteers have shown that the new formulation of this drug is outstanding due a twofold better lung deposition, compared to the reference medicine. The aim of this study was to evaluate the efficacy and safety of the new formulation of fluticasone propionate administered through new generation cyclohaler (CNG), compared to original fluticasone administered through dry powder inhaler (DPI) in patients with chronic moderate asthma.MATERIAL END METHODS: The study included 457 patients. 376 subjects were randomized to one out of the three groups: 127 subjects — to the group treated with the new formulation of fluticasone at a dose of 125 μg BID, 125 subjects — to the group treated with new formulation of fluticasone at a dose of 250 μg BID, and 124 subjects — to the group treated with the reference drug — fluticasone DPI 500 μg BID. At the beginning of the study, the groups did not differ in demographical or clinical aspects. Active therapy lasted 12 weeks. The primary endpoint was a mean change in morning PEF during a 12-week course of therapy (ΔmPEF of 15 L/min was considered as statistically significant). Additionally, other functional parameters of the respiratory system — clinical symptoms and the use of rescue drugs were studied. During the whole study the safety of patients was monitored by recording adverse events; in addition, a systemic exposure to fluticasone was evaluated by testing the changes of cortisol in serum and in a 24-hour collection of urine in a subgroup consisting of 45 patients. Statistical analysis was conducted on both groups: intention-to-treat (ITT) and per protocol (PP).RESULTS: In PP as well as in ITT analysis, a mean change in morning PEF at the end of the therapy in comparison with the initial period was statistically significant in all therapeutic groups. The efficacy of the treatment with fluticasone at doses of 125 μg BID and 250 μg and the reference medicines did not differ statistically significantly after a 12-week course of therapy or during the whole period of treatment. During the study, significant improvement in the range of other functional parameters such as evening PEF, FEV1, clinical symptoms and the use of rescue drugs was observed in all therapeutic groups, without significant differences in efficacy between the study groups. The comparison of efficacy of fluticasone at a dose of 125 μg BID with the generic product at a dose of 250 μg BID showed a weak dose-response relationship concerning the change in morning PEF, which arises from the almost flat dose-response curve in the range of medium and high doses for this drug. No significant quantitative or qualitative differences were shown between the groups in the recorded adverse events, qualified as related to treatment with fluticasone. There were no significant changes revealed in cortisol concentration in serum or in a 24-hour collection of urine between the initial level and the final visit in any of the groups.CONCLUSIONS: Fluticasone administered through the new generation cyclohaler, compared to original fluticasone DPI, allows a twofold reduction in drug dose, retaining in new formulation clinical efficacy that corresponds to the reference drug at twice the dose. New formulation of fluticasone administered through the new generation cyclohaler has a safety profile clinically comparable to the reference drug

Endoplasmic reticulum dysfunction in neurological disease.

Endoplasmic reticulum (ER) dysfunction might have an important part to play in a range of neurological disorders, including cerebral ischaemia, sleep apnoea, Alzheimer's disease, multiple sclerosis, amyotrophic lateral sclerosis, the prion diseases, and familial encephalopathy with neuroserpin inclusion bodies. Protein misfolding in the ER initiates the well studied unfolded protein response in energy-starved neurons during stroke, which is relevant to the toxic effects of reperfusion. The toxic peptide amyloid β induces ER stress in Alzheimer's disease, which leads to activation of similar pathways, whereas the accumulation of polymeric neuroserpin in the neuronal ER triggers a poorly understood ER-overload response. In other neurological disorders, such as Parkinson's and Huntington's diseases, ER dysfunction is well recognised but the mechanisms by which it contributes to pathogenesis remain unclear. By targeting components of these signalling responses, amelioration of their toxic effects and so the treatment of a range of neurodegenerative disorders might become possible

Lack of correlation between exhaled nitric oxide (eNO) and clinical indicators of the disease activity and quality of life in mild and moderate asthmatics

The measurements of exhaled nitric oxide (eNO) are simple and useful method of assessment of inflammation in asthmatics’ airways. One of the causes of its limited application in clinical practice is a number of factors influencing the results of measurements. The aim of the study was to determine the usefulness of eNO measurements in assessing the inflammation in a heterogeneous, in relation to atopic and smoking status, group of patients. Materials and methods: 120 subjects suspected of having asthma participated in this study. During 2 weeks the patients noted daily asthma symptoms and daily use of rescue medication. After 14 days health related quality of life (HRQL) was determined by means of Asthma Quality of Life Questionnaire (AQLQ), eNO levels were measured and airways reversibility test was performed. Results: Preliminary diagnosis of asthma was confi rmed in 84 patients on the basis of positive result of airways reversibility test. Among them, 21 subjects (25%) were smokers and 60 (71.4%) were atopic. No correlation was found between eNO and daily asthma symptom score, daily use of rescue medication, percent of airway reversibility after β2-agonist and HRQL. Conclusion: eNO measurements in a heterogeneous, in relation to atopic and smoking status, group of patients are of limited value in clinical assessment of asthma activity

The TRiC/CCT chaperone is implicated in Alzheimer's disease based on patient GWAS and an RNAi screen in Aβ-expressing Caenorhabditis elegans.

The human Aβ peptide causes progressive paralysis when expressed in the muscles of the nematode worm, C. elegans. We have exploited this model of Aβ toxicity by carrying out an RNAi screen to identify genes whose reduced expression modifies the severity of this locomotor phenotype. Our initial finding was that none of the human orthologues of these worm genes is identical with the genome-wide significant GWAS genes reported to date (the "white zone"); moreover there was no identity between worm screen hits and the longer list of GWAS genes which included those with borderline levels of significance (the "grey zone"). This indicates that Aβ toxicity should not be considered as equivalent to sporadic AD. To increase the sensitivity of our analysis, we then considered the physical interactors (+1 interactome) of the products of the genes in both the worm and the white+grey zone lists. When we consider these worm and GWAS gene lists we find that 4 of the 60 worm genes have a +1 interactome overlap that is larger than expected by chance. Two of these genes form a chaperonin complex, the third is closely associated with this complex and the fourth gene codes for actin, the major substrate of the same chaperonin

p53 and translation attenuation regulate distinct cell cycle checkpoints during endoplasmic reticulum (ER) stress.

Cell cycle checkpoints ensure that proliferation occurs only under permissive conditions, but their role in linking nutrient availability to cell division is incompletely understood. Protein folding within the endoplasmic reticulum (ER) is exquisitely sensitive to energy supply and amino acid sources because deficiencies impair luminal protein folding and consequently trigger ER stress signaling. Following ER stress, many cell types arrest within the G(1) phase, although recent studies have identified a novel ER stress G(2) checkpoint. Here, we report that ER stress affects cell cycle progression via two classes of signal: an early inhibition of protein synthesis leading to G(2) delay involving CHK1 and a later induction of G(1) arrest associated both with the induction of p53 target genes and loss of cyclin D(1). We show that substitution of p53/47 for p53 impairs the ER stress G(1) checkpoint, attenuates the recovery of protein translation, and impairs induction of NOXA, a mediator of cell death. We propose that cell cycle regulation in response to ER stress comprises redundant pathways invoked sequentially first to impair G(2) progression prior to ultimate G(1) arrest

Suppression of Aβ toxicity by puromycin-sensitive aminopeptidase is independent of its proteolytic activity.

The accumulation of β-amyloid (Aβ) peptide in the brain is one of the pathological hallmarks of Alzheimer's disease and is thought to be of primary aetiological significance. In an unbiased genetic screen, we identified puromycin-sensitive aminopeptidase (PSA) as a potent suppressor of Aβ toxicity in a Drosophila model system. We established that coexpression of Drosophila PSA (dPSA) in the flies' brains improved their lifespan, protected against locomotor deficits, and reduced brain Aβ levels by clearing the Aβ plaque-like deposits. However, confocal microscopy and subcellular fractionation of amyloid-expressing 7PA2 cells demonstrated that PSA localizes to the cytoplasm. Therefore, PSA and Aβ are unlikely to be in the same cellular compartment; moreover, when we artificially placed them in the same compartment in flies, we could not detect a direct epistatic interaction. The consequent hypothesis that PSA's suppression of Aβ toxicity is indirect was supported by the finding that Aβ is not a proteolytic substrate for PSA in vitro. Furthermore, we showed that the enzymatic activity of PSA is not required for rescuing Aβ toxicity in neuronal SH-SY5Y cells. We investigated whether the stimulation of autophagy by PSA was responsible for these protective effects. However PSA's promotion of autophagosome fusion with lysosomes required proteolytic activity and so its effect on autophagy is not identical to its protection against Aβ toxicity

Sterol metabolism regulates neuroserpin polymer degradation in the absence of the unfolded protein response in the dementia FENIB.

Mutants of neuroserpin are retained as polymers within the endoplasmic reticulum (ER) of neurones to cause the autosomal dominant dementia familial encephalopathy with neuroserpin inclusion bodies or FENIB. The cellular consequences are unusual in that the ordered polymers activate the ER overload response (EOR) in the absence of the canonical unfolded protein response. We use both cell lines and Drosophila models to show that the G392E mutant of neuroserpin that forms polymers is degraded by UBE2j1 E2 ligase and Hrd1 E3 ligase while truncated neuroserpin, a protein that lacks 132 amino acids, is degraded by UBE2g2 (E2) and gp78 (E3) ligases. The degradation of G392E neuroserpin results from SREBP-dependent activation of the cholesterol biosynthetic pathway in cells that express polymers of neuroserpin (G392E). Inhibition of HMGCoA reductase, the limiting enzyme of the cholesterol biosynthetic pathway, reduced the ubiquitination of G392E neuroserpin in our cell lines and increased the retention of neuroserpin polymers in both HeLa cells and primary neurones. Our data reveal a reciprocal relationship between cholesterol biosynthesis and the clearance of mutant neuroserpin. This represents the first description of a link between sterol metabolism and modulation of the proteotoxicity mediated by the EOR

Characterisation of serpin polymers in vitro and in vivo

Neuroserpin is a member of the serine protease inhibitor or serpin superfamily of proteins. It is secreted by neurones and plays an important role in the regulation of tissue plasminogen activator at the synapse. Point mutations in the neuroserpin gene cause the autosomal dominant dementia familial encephalopathy with neuroserpin inclusion bodies or FENIB. This is one of a group of disorders caused by mutations in the serpins that are collectively known as the serpinopathies. Others include alpha(1)-antitrypsin deficiency and deficiency of C1 inhibitor, antithrombin and alpha(1)-antichymotrypsin. The serpinopathies are characterised by delays in protein folding and the retention of ordered polymers of the mutant serpin within the cell of synthesis. The clinical phenotype results from either a toxic gain of function from the inclusions or a loss of function, as there is insufficient protease inhibitor to regulate important proteolytic cascades. We describe here the methods required to characterise the polymerisation of neuroserpin and draw parallels with the polymerisation of alpha(1)-antitrypsin. It is important to recognise that the conditions in which experiments are performed will have a major effect on the findings. For example, incubation of monomeric serpins with guanidine or urea will produce polymers that are not found in vivo. The characterisation of the pathological polymers requires heating of the folded protein or alternatively the assessment of ordered polymers from cell and animal models of disease or from the tissues of humans who carry the mutation. (C) 2010 Elsevier Inc. All rights reserved