The isoprenoid end product N6-isopentenyladenosine reduces inflammatory response through the inhibition of the NF\uce\ubaB and STAT3 pathways in cystic fibrosis cells

N6-isopentenyladenosine (iPA) is an intermediate of the mevalonate pathway that exhibits various anti-cancer effects. However, studies on its anti-inflammatory activity are scarce and underlying molecular mechanisms are unknown. Therefore, we aimed to investigate the ability of iPA to exert anti-inflammatory effects in the human cystic fibrosis (CF) cell model of exacerbated inflammation. TNF alpha-stimulated CF cells CuFi-1 and its normal counterpart NuLi-1 were pre-treated with increasing concentrations of iPA and cell viability and proliferation were assessed by MTT and BrdU assays. The effect of iPA on IL-8 and RANTES secretion was determined by ELISA, and the activation and expression of signaling molecules and selenoproteins were studied by Western blot. To assess the direct effect of iPA on NF kappa B activity, luciferase assay was performed on TNF alpha-stimulated HEK293/T cells transfected with a NF kappa B reporter plasmid. We demonstrated for the first time that iPA prevents IL-8 and RANTES release in TNF alpha-stimulated CF cells and this effect is mediated by increasing the expression of the direct NF kappa B inhibitor I kappa B alpha and decreasing the levels of STAT3. Consistent with this, we showed that iPA inhibited TNF alpha-mediated NF kappa B activation in HEK/293T cells. Finally, we also found that iPA improved the levels of glutathione peroxidase 1 and thioredoxin reductase 1 only in CF cells suggesting its ability to maintain sufficient expression of these anti-oxidant selenoproteins. Our findings indicate that iPA can exert anti-inflammatory activity especially in the cases of excessive inflammatory response as in CF

Structure-activity relationships, biological evaluation and structural studies of novel pyrrolonaphthoxazepines as antitumor agents

Microtubule-targeting agents (MTAs) are a class of clinically successful anti-cancer drugs. The emergence of multidrug resistance to MTAs imposes the need for developing new MTAs endowed with diverse mechanistic properties. Benzoxazepines were recently identified as a novel class of MTAs. These anticancer agents were thoroughly characterized for their antitumor activity, although, their exact mechanism of action remained elusive. Combining chemical, biochemical, cellular, bioinformatics and structural efforts we developed improved pyrrolonaphthoxazepines antitumor agents and their mode of action at the molecular level was elucidated. Compound 6j, one of the most potent analogues, was confirmed by X-ray as a colchicine-site MTA. A comprehensive structural investigation was performed for a complete elucidation of the structure-activity relationships. Selected pyrrolonaphthoxazepines were evaluated for their effects on cell cycle, apoptosis and differentiation in a variety of cancer cells, including multidrug resistant cell lines. Our results define compound 6j as a potentially useful optimized hit for the development of effective compounds for treating drug-resistant tumors.This work was supported in part by a grant from the Swiss National Science Foundation (31003A_166608; to M.O.S), grant BFU2016-75319-R (AEI/FEDER, EU) from Ministerio de Economia y Competitividad, Blueprint 282510, AIRC-17217. The authors acknowledge networking contribution by the COST Action CM1407 “Challenging organic syntheses inspired by nature - from natural products chemistry to drug discovery” (to M.O.S. and J.F.D.) and the COST Action EPICHEMBIO CM-1406 (to L.A. and G.C.). This work has also received partial funding from the European Union’s Horizon 2020 (EU) research and innovation programme under the Marie Sklodowska-Curie grant agreement No 721906. Finally, this work was partially funded by MIUR-PRIN project n. 2015Y3C5KP (to L.M.)

Design and characterization of DPI (dry powder inhaler) for the pulmonary delivery of anti-inflammatory and antibiotic drugs in the treatment of cystic fibrosis disease

2010 - 2011The aim of the present PhD project was to design inhalable powder-based formulations for pharmaceutical products that may improve the treatment of pulmonary diseases, mainly cystic fibrosis, and may be easier for patients to use. Particularly, the present project aims to supply CF patients with flavonoids (Naringin) and aminoglycosides (Gentamicin sulfate) in a respirable form as a valid alternative over more conventional (oral or parenteral) anti-inflammatory and antibiotic therapy. As a matter of fact, in CF epithelial cells, antioxidant defense systems appear to be defective in their ability to control the amount of ROS produced and over abundance of ROS may cause tissue injury-events and modify intracellular signalling pathways leading to enhanced inflammatory processes, typical of CF airways. Overall, evidence suggests improved CFTR function in vitro when flavonoids, such as genistein, are used. For chronic Pseudomonas aeruginosa (Pa) infections in CF, gentamicin given by pulmonary route may plays important role. In fact, it was observed daily inhalation of some aminoglycosides from nebulized solution delays the acquisition of chronic Pa infections and decreases CF progression. The project address a number of the key features that are outstanding in inhaled delivery, mainly - characteristic of the active drug; - properties of the drug formulation, particularly powder flow, particle size, shape, surface properties and drug/carrier interaction; - consistent dose delivery and high proportion of dose getting to the lung; - performance of the inhaler device, including aerosol generation and delivery. A balance among these characteristics is necessary in the design of a drug formulation intended for pulmonary administration. Utilizing proven (Spray-drying) or innovative (Supercritical Assisted Atomization) technology, stable and micronized powders usefull for dry powder inhaler (DPI) production have been developed. Moreover, the research has been based on in vitro product test methods to evaluate the health effects of produced powders and their aerodynamic behaviour through the pulmonary system. Optimized stability and bioavailability of the selected drugs, the achieving of therapeutically effective concentrations for the pulmonary care of cystic fibrosis have been other goals of the research. Technologies and products that the research is aimed to develop would be of interest to a number of pharmaceutical companies either in the respiratory area or trying to get a toehold in this market. Specific objectives of this research have been: design and development of Dry Powder Inhalers (DPIs) containing flavonoids (Naringin) or aminoglycoside antibiotics (Gentamicin sulfate) micronized powder by spray drying production or by Supercritical Assisted Atomization (SAA); optimization of the aerodynamic characteristics of the powders, through the use of excipients (amino acids) not toxic for lung but able to improve the powder flow properties and dispersion which, in turn, may increase lung deposition of the drugs; in vitro evaluation of the biological activity of the engineered particles on a model of bronchial epithelial cell lines from patients with cystic fibrosis (CuFi1, F508del/F508del CFTR), in comparison to the activity of the same products on normal bronchial epithelial cell lines (NuLi1). (edited by author)X n.s

Leucine enhances aerosol performance of Naringin dry powder and its activity on cystic fibrosis airway epithelial cells

The effect of different amino acids (AAs) on the aerosol performance of N spray-dried powders was studied. Morphology, size distribution, density, dissolution rate were evaluated and correlated to process parameters. The aerosol performance was analyzed by both Single Stage Glass Impinger and Andersen Cascade Impactor. Results indicated that powders containing 5% (w/w) of leucine, proline or histidine and dried from 3:7 ethanol/water feeds showed very satisfying aerodynamic properties with fine particle fraction > 60%. Both neat N (raw and spray-dried) and N-leu1 dry-powder showing good aerodynamic properties were tested in cystic fibrosis (CF) and normal bronchial epithelial cells. Cell proliferation and expression levels of the key enzymes of the NF-κB and MAPK/ERK pathways, overactivated in CF cell lines, were evaluated. N-leu1 was able to significantly inhibit the expression levels of IKKα, IKKβ, as well as of the direct NF-κB inhibitor, IκBα. In addition N-Leu1 inhibited phosphorylation of ERK1/2 kinase and did not reduce cell proliferation as observed for the neat raw drug. Leucine co-spray-dried with the drug improved both aerodynamic properties and in vitro pharmacological activity of Naringin. The optimized N-Leu formulation as dry powder is potentially able to reduce hyperinflammatory status associated to CF

PETrA: A software-based tool for estimating the energy profile of android applications

Energy efficiency is a vital characteristic of any mobile application, and indeed is becoming an important factor for user satisfaction. For this reason, in recent years several approaches and tools for measuring the energy consumption of mobile devices have been proposed. Hardware-based solutions are highly precise, but at the same time they require costly hardware toolkits. Model-based techniques require a possibly difficult calibration of the parameters needed to correctly create a model on a specific hardware device. Finally, software-based solutions are easier to use, but they are possibly less precise than hardware-based solution. In this demo, we present PETrA, a novel software-based tool for measuring the energy consumption of Android apps. With respect to other tools, PETrA is compatible with all the smartphones with Android 5.0 or higher, not requiring any device specific energy profile. We also provide evidence that our tool is able to perform similarly to hardware-based solutions

Software-based energy profiling of Android apps: Simple, efficient and reliable?

Modeling the power profile of mobile applications is a crucial activity to identify the causes behind energy leaks. To this aim, researchers have proposed hardware-based tools as well as model-based and software-based techniques to approximate the actual energy profile. However, all these solutions present their own advantages and disadvantages. Hardware-based tools are highly precise, but at the same time their use is bound to the acquisition of costly hardware components. Model-based tools require the calibration of parameters needed to correctly create a model on a specific hardware device. Software-based approaches do not need any hardware components, but they rely on battery measurements and, thus, they are hardware-assisted. These tools are cheaper and easier to use than hardware-based tools, but they are believed to be less precise. In this paper, we take a deeper look at the pros and cons of software-based solutions investigating to what extent their measurements depart from hardware-based solutions. To this aim, we propose a software-based tool named PETRA that we compare with the hardware-based MONSOON toolkit on 54 Android apps. The results show that PETRA performs similarly to MONSOON despite not using any sophisticated hardware components. In fact, in all the apps the mean relative error with respect to MONSOON is lower than 0.05. Moreover, for 95% of the analyzed methods the estimation error is within 5% of the actual values measured using the hardware-based toolkit

Gentamicin and leucine inhalable powder: What about antipseudomonal activity and permeation through cystic fibrosis mucus?

The aim of this study was to evaluate the permeation properties of gentamicin (G) in a novel dry powder form for inhalation through an artificial mucus model. Moreover, since respiratory infections sustained by Pseudomonas are a major cause of sickness and death in CF patients, the susceptibility of P. aeruginosa to engineered G powders was investigated. Micronized G and G/leucine (85:15) formulations were produced by co-spray-drying, using process parameters and conditions previously set. Powders were characterized in terms of yield, drug content and aerodynamic profiles, analyzed by Andersen Cascade Impactor. Different mucus models were prepared, showing composition and viscosity similar to those of the native CF mucus. To investigate the impact on drug permeation, Franz-type vertical diffusion cells were used; the powders were applied directly on a synthetic membrane with or without the interposition of the artificial mucus layer. In buffer, gentamicin showed a diffusion controlled release; the presence of leucine reduced powder wettability and, consequently, the permeation rate. Otherwise, mucus delayed drug permeation from both G and G/leucine formulations, with a faint influence of the aminoacid. Antimicrobial tests revealed that G/leu engineered particles are able to preserve the antipseudomonal activity, even in presence of the mucus