Pseudomonas aeruginosa induces p38MAP kinase-dependent IL-6 and CXCL8 release from bronchial epithelial cells via a Syk kinase pathway.

Pseudomonas aeruginosa (Pa) infection is a major cause of airway inflammation in immunocompromised and cystic fibrosis (CF) patients. Mitogen-activated protein (MAP) and tyrosine kinases are integral to inflammatory responses and are therefore potential targets for novel anti-inflammatory therapies. We have determined the involvement of specific kinases in Pa-induced inflammation. The effects of kinase inhibitors against p38MAPK, MEK 1/2, JNK 1/2, Syk or c-Src, a combination of a p38MAPK with Syk inhibitor, or a novel narrow spectrum kinase inhibitor (NSKI), were evaluated against the release of the proinflammatory cytokine/chemokine, IL-6 and CXCL8 from BEAS-2B and CFBE41o- epithelial cells by Pa. Effects of a Syk inhibitor against phosphorylation of the MAPKs were also evaluated. IL-6 and CXCL8 release by Pa were significantly inhibited by p38MAPK and Syk inhibitors (p<0.05). Phosphorylation of HSP27, but not ERK or JNK, was significantly inhibited by Syk kinase inhibition. A combination of p38MAPK and Syk inhibitors showed synergy against IL-6 and CXCL8 induction and an NSKI completely inhibited IL-6 and CXCL8 at low concentrations. Pa-induced inflammation is dependent on p38MAPK primarily, and Syk partially, which is upstream of p38MAPK. The NSKI suggests that inhibiting specific combinations of kinases is a potent potential therapy for Pa-induced inflammation.

Discovery of novel benzothienoazepine derivatives as potent inhibitors of respiratory syncytial virus.

The development of novel non-nucleoside inhibitors of the RSV polymerase complex is of significant clinical interest. Compounds derived from the benzothienoazepine core, such as AZ-27, are potent inhibitors of RSV viruses of the A-subgroup, but are only moderately active against the B serotype and as yet have not demonstrated activity in vivo. Herein we report the discovery of several novel families of C-2 arylated benzothienoazepine derivatives that are highly potent RSV polymerase inhibitors and reveal an exemplary structure, compound 4a, which shows low nanomolar activity against both RSV A and B viral subtypes. Furthermore, this compound is effective at suppressing viral replication, when administered intranasally, in a rodent model of RSV infection. These results suggest that compounds belonging to this chemotypes have the potential to provide superior anti-RSV agents than those currently available for clinical use.

Influence of mouthpiece geometry on the aerosol delivery performance of a dry powder inhaler.

PURPOSE: To investigate the influence of mouthpiece geometry on the amount of throat deposition and device retention produced using a dry powder inhaler (Aerolizer), along with the subsequent effect on the overall inhaler performance. MATERIALS AND METHODS: Computational Fluid Dynamics analysis of the flowfield generated in the Aerolizer with various modified mouthpiece geometries (including cylindrical, conical and oval designs) was used in conjunction with experimental dispersions of mannitol powder using a multi-stage liquid impinger to determine how the overall inhaler performance varied as the mouthpiece geometry was modified. RESULTS: Geometry of the inhaler mouthpiece had no effect on device retention or the inhaler dispersion performance. In contrast, the mouthpiece geometry strongly affected the amount of throat deposition by controlling the axial component of the exit air flow velocity. The radial motion of the emitted aerosol jet was found to have little effect on throat deposition in representative mouth-throat models. Despite the reduced throat deposition, there was no difference in the overall inhaler performance. CONCLUSIONS: For cases where low throat deposition is a key design parameter, this study demonstrates that the amount of throat deposition can be reduced by making minor modifications to the inhaler mouthpiece design.

Effect of design on the performance of a dry powder inhaler using computational fluid dynamics. Part 2: Air inlet size.

This study investigates the effect of air inlet size on (i) the flowfield generated in a dry powder inhaler, and (ii) the device-specific resistance, and the subsequent effect on powder deagglomeration. Computational fluid dynamics (CFD) analysis was used to simulate the flowfield generated in an Aerolizer with different air inlet sizes at 30, 45, and 60 l/min. Dispersion performance of the modified inhalers was measured using mannitol powder and a multistage liquid impinger at the same flow rates. The air inlet size had a varying effect on powder dispersion depending on the flow rate. At low flow rates (30 and 45 l/min), reducing the air inlet size increased the inhaler dispersion performance by increasing the flow turbulence and particle impaction velocities above their critical levels for maximal powder dispersion. At 60 l/min, reducing the air inlet size reduced the inhaler dispersion performance by releasing a large amount of powder from the device before the turbulence levels and particle impaction velocities could be fully developed. The results demonstrate that the maximal inhaler dispersion performance can be predicted if details of the device flowfield are known.

Influence of air flow on the performance of a dry powder inhaler using computational and experimental analyses.

PURPOSE: The aims of the study are to analyze the influence of air flow on the overall performance of a dry powder inhaler (Aerolizer and to provide an initial quantification of the flow turbulence levels and particle impaction velocities that maximized the inhaler dispersion performance. METHODS: Computational fluid dynamics (CFD) analysis of the flow field in the Aerolizer, in conjunction with experimental dispersions of mannitol powder using a multistage liquid impinger, was used to determine how the inhaler dispersion performance varied as the device flow rate was increased. RESULTS: Both the powder dispersion and throat deposition were increased with air flow. The capsule retention was decreased with flow, whereas the device retention first increased then decreased with flow. The optimal inhaler performance was found at 65 l min(-1) showing a high fine particle fraction (FPF) of 63 wt.% with low throat deposition (9.0 wt.%) and capsule retention (4.3 wt.%). Computational fluid dynamics analysis showed that at the critical flow rate of 65 l min(-1), the volume-averaged integral scale strain rate (ISSR) was 5,400 s(-1), and the average particle impaction velocities were 12.7 and 19.0 m s(-1) at the inhaler base and grid, respectively. Correlations between the device flow rate and (a) the amount of throat deposition and (b) the capsule emptying times were also developed. CONCLUSIONS: The use of CFD has provided further insight into the effect of air flow on the performance of the Aerolizer. The approach of using CFD coupled with powder dispersion is readily applicable to other dry powder inhalers (DPIs) to help better understand their performance optimization.

The role of capsule on the performance of a dry powder inhaler using computational and experimental analyses.

PURPOSE: To study the fundamental effects of the spinning capsule on the overall performance of a dry powder inhaler (Aerolizer). METHODS: The capsule motion was visualized using high-speed photography. Computational fluid dynamics (CFD) analysis was performed to determine the flowfield generated in the device with and without the presence of different sized capsules at 60 l min(-1). The inhaler dispersion performance was measured with mannitol powder using a multistage liquid impinger at the same flowrate. RESULTS: The capsule size (3, 4, and 5) was found to make no significant difference to the device flowfield, the particle-device impaction frequency, or the dispersion performance of the inhaler. Reducing the capsule size reduced only the capsule retention by 4%. In contrast, without the presence of the spinning capsule, turbulence levels were increased by 65%, FPF(Em) (wt% particles < or =6.8 microm in the aerosol referenced against the amount of powder emitted from the device) increased from 59% to 65%, while particle-mouthpiece impaction decreased by 2.5 times. When the powder was dispersed from within compared to from outside the spinning capsule containing four 0.6 mm holes at each end, the FPF(Em) was increased significantly from 59% to 76%, and the throat retention was dropped from 14% to 6%. CONCLUSIONS: The presence, but not the size, of a capsule has significant effects on the inhaler performance. The results suggested that impaction between the particles and the spinning capsule does not play a major role in powder dispersion. However, the capsule can provide additional strong mechanisms of deagglomeration dependent on the size of the capsule hole.

Effect of design on the performance of a dry powder inhaler using computational fluid dynamics. Part 1: Grid structure and mouthpiece length.

This study investigates (1) the effect of modifying the design of a dry powder inhaler on the device performance, and (2) which design features significantly contribute to overall inhaler performance. Computational Fluid Dynamics (CFD) analysis was performed to determine how the flowfield generated in an Aerolizer at 60 l min(-1) varied when the inhaler grid and mouthpiece were modified. The computational models were validated by Laser Doppler Velocimetry (LDV). Dispersion performance of the modified inhalers was measured with a mannitol powder using a multistage liquid impinger at 60 l min(-1). The inhaler grid was found to significantly affect the performance of the Aerolizer. As the grid voidage was increased, the amount of powder retained in the device doubled (due to increased tangential flow of particles in the inhaler mouthpiece) and the FPF(Loaded) was reduced from 57 to 44% (due to increased mouthpiece retention). The length of the mouthpiece played a lesser role on the inhaler performance, having no significant effect on the flowfield generated in the devices. In summary, the performance of a dry powder inhaler can be affected by simple design changes. CFD, coupled with experimental results, provides a rational basis for understanding the performance difference.