Inflammation-induced microsomal prostaglandin E synthase-1 (mPGES-1) is the terminal enzyme that synthesizes prostaglandin E
2 (PGE
2) downstream of cyclooxygenase-2 (COX-2). The efficacy of ...nonsteroidal anti-inflammatory drugs and COX-2 inhibitors in the treatment of the signs and symptoms of osteoarthritis, rheumatoid arthritis and inflammatory pain, largely attributed to the inhibition of PGE
2 synthesis, provides a rationale for exploring mPGES-1 inhibition as a potential novel therapy for these diseases. Toward this aim, we identified PF-9184 as a novel mPGES-1 inhibitor. PF-9184 potently inhibited recombinant human (rh) mPGES-1 (IC
50
=
16.5
±
3.8
nM), and had no effect against rhCOX-1 and rhCOX-2 (>6500-fold selectivity). In inflammation and clinically relevant biological systems, mPGES-1 expression, like COX-2 expression was induced in cell context- and time-dependent manner, consistent with the kinetics of PGE
2 synthesis. In rationally designed cell systems ideal for determining direct effects of the inhibitors on mPGES-1 function, but not its expression, PF-9184 inhibited PGE
2 synthesis (IC
50 in the range of 0.5–5
μM in serum-free cell and human whole blood cultures, respectively) while sparing the synthesis of 6-keto-PGF
1α (PGF
1α) and PGF
2α. In contrast, as expected, the selective COX-2 inhibitor, SC-236, inhibited PGE
2, PGF
1α and PGF
2α synthesis. This profile of mPGES-1 inhibition, distinct from COX-2 inhibition in cells, validates mPGES-1 as an attractive target for therapeutic intervention.
p38α activation of multiple effectors may underlie the failure of global p38α inhibitors in clinical trials. A unique inhibitor (CDD-450) was developed that selectively blocked p38α activation of the ...proinflammatory kinase MK2 while sparing p38α activation of PRAK and ATF2. Next, the hypothesis that the p38α-MK2 complex mediates inflammasome priming cues was tested. CDD-450 had no effect on NLRP3 expression, but it decreased IL-1β expression by promoting IL-1β mRNA degradation. Thus, IL-1β is regulated not only transcriptionally by NF-κB and posttranslationally by the inflammasomes but also posttranscriptionally by p38α-MK2. CDD-450 also accelerated TNF-α and IL-6 mRNA decay, inhibited inflammation in mice with cryopyrinopathy, and was as efficacious as global p38α inhibitors in attenuating arthritis in rats and cytokine expression by cells from patients with cryopyrinopathy and rheumatoid arthritis. These findings have clinical translation implications as CDD-450 offers the potential to avoid tachyphylaxis associated with global p38α inhibitors that may result from their inhibition of non-MK2 substrates involved in antiinflammatory and housekeeping responses.
A series of pyrazole inhibitors of p38 mitogen-activated protein (MAP) kinase were designed using a binding model based on the crystal structure of 1 (SC-102) bound to p38 enzyme. New chemistry using ...dithietanes was developed to assemble nitrogen-linked substituents at the 5-position of pyrazoles. Calculated log D was used in tandem with structure-based design to guide medicinal chemistry strategy and improve the in vivo activity of a series of molecules. The crystal structure of an optimized inhibitor, 4 (SC-806), in complex with p38 enzyme was obtained to confirm the hypothesis that the addition of a basic nitrogen to the molecule induces an interaction with Asp112 of p38α. A compound identified from this series was efficacious in an animal model of rheumatic disease.
In this report, we show that apoptosis signal-regulating kinase 1
−/− (ASK1 KO) mice were resistant to inflammatory arthritis induced in the K/BxN serum transfer model of rheumatoid arthritis (RA). ...The p38 inhibitor, SD-0006 was administered to wild type (WT) mice as a comparator. Both ASK1 KO and p38 inhibition resulted in marked attenuation of edema, cartilage damage, bone resorption, and general inflammatory responses. Transcriptional profiling of mRNA prepared from paw tissue demonstrated that the production of many proinflammatory genes including cytokines, chemokines, and extracellular matrix degradative enzymes were maintained at basal levels by either ASK1 KO or prophylactic p38 MAPK inhibition. In the mouse whole blood (MWB) assay, tumor necrosis factor-α (TNF-α)-induced KC and CCL2 levels and also LPS-induced interleukin-6 (IL-6), CCL2, and KC levels in MWB from ASK1 KO were significantly lower than those from WT. Furthermore, both p38 and JNK were activated by TNF-α in human synovial fibroblasts isolated from RA patients (RASF). SD-0006 or SP600125, a JNK inhibitor, partially blocked the elevation of IL-6 production in RASF following stimulation with TNF-α. In contrast, dual inhibition with both p38/JNK inhibitors almost completely abolished TNF-α-induced IL-6 production from these cells. Ablation of ASK1 expression in RASF using siRNA for ASK1 resulted in inhibition of TNF-α-induced IL-6 and PGE
2 production. This study is the first to suggest that ASK1 is critical for the development of RA and that ASK1 may be involved in the production of proinflammatory mediators in response to TNF-α stimulation in the RA joint.
► ASK1 KO mice are resistant to K/BxN serum-induced arthritis.► LPS-induced chemokine level in whole blood from ASK1KO was lower than that from WT.► Both p38 and JNK inhibition completely inhibited TNF α-induced IL6 production in RASF.► Ablation of ASK1 expression in RASF resulted in TNFα-induced IL6 and PGE2 production.
The contribution of inducible nitric oxide synthase
(iNOS) to oxidative/nitrative stress is well-documented in inflammation, but difficult to quantify. Using a novel, recently developed assay for ...3-nitrotyrosine (3-NT), we characterized iNOS activity and its inhibition in preclinical models of inflammation. In particular, we utilized the 3-NT assay to assess the role of iNOS in the disease pathology as well as for proof of pharmacology of iNOS inhibitors in an acute endotoxin challenge model, in models of rheumatoid arthritis (RA) such as rat adjuvant- and collagen-induced arthritis (AIA and CIA) and a model of osteoarthritis (OA) such as rat sodium monoiodoacetate-induced arthritis (MIA). Quantification of nitrotyrosine was performed using immuno-affinity 2-D LC–MS/MS assay. This assay is a very specific and reproducible and is amenable to a number of biological fluids. Plasma levels of 3-NT were significantly elevated in an acute model of inflammation (rat LPS) and in models of rheumatoid arthritis (adjuvant- and collagen-induced arthritis), and osteoarthritis (monoiodoacetate-induced arthritis). Plasma 3-NT correlated with the severity of the inflammatory response; thus, a 20-fold increase was observed in the rat LPS model, a 10-fold increase in AIA, and only a 2.5-fold elevation in CIA. Pharmacological intervention with iNOS inhibitors decreased 3-NT levels and associated pathology. 3-NT determination allowed for better elucidation of the role of iNOS in RA and OA disease patholog
y and provided proof of pharmacology for NOS inhibitors in animal models of RA and OA.
Activation of the p38 kinase pathway in immune cells leads to the transcriptional and translational regulation of proinflammatory cytokines. Mitogen-activated protein kinase-activated protein kinase ...2 (MK2), a direct downstream substrate of p38 kinase, regulates lipopolysaccharide (LPS)-stimulated tumor necrosis factor alpha (TNFalpha) and interleukin-6 (IL-6) production through modulating the stability and translation of these mRNAs. Developing small-molecule inhibitors of MK2 may yield anti-inflammatory efficacy with a different safety profile relative to p38 kinase inhibitors. This article describes the pharmacologic properties of a benzothiophene MK2 inhibitor, PF-3644022 (10R)-10-methyl-3-(6-methylpyridin-3-yl)-9,10,11,12-tetrahydro-8H-1,4diazepino5',6':4,5thieno3,2-fquinolin-8-one. PF-3644022 is a potent freely reversible ATP-competitive compound that inhibits MK2 activity (K(i) = 3 nM) with good selectivity when profiled against 200 human kinases. In the human U937 monocytic cell line or peripheral blood mononuclear cells, PF-3644022 potently inhibits TNFalpha production with similar activity (IC(50) = 160 nM). PF-3644022 blocks TNFalpha and IL-6 production in LPS-stimulated human whole blood with IC(50) values of 1.6 and 10.3 microM, respectively. Inhibition of TNFalpha in U937 cells and blood correlates closely with inhibition of phospho-heat shock protein 27, a target biomarker of MK2 activity. PF-3644022 displays good pharmacokinetic parameters in rats and is orally efficacious in both the rat acute LPS-induced TNFalpha model and the chronic streptococcal cell wall-induced arthritis model. Dose-dependent inhibition of TNFalpha production in the acute model and inhibition of paw swelling in the chronic model is observed with ED(50) values of 6.9 and 20 mg/kg, respectively. PF-3644022 efficacy in the chronic inflammation model is strongly correlated with maintaining a C(min) higher than the EC(50) measured in the rat LPS-induced TNFalpha model.
Physics at the e+e- linear collider Moortgat-Pick, G.; Baer, H.; Battaglia, M. ...
The European physical journal. C, Particles and fields,
08/2015, Letnik:
75, Številka:
8
Journal Article
Recenzirano
Odprti dostop
A comprehensive review of physics at an
e
+
e
-
linear collider in the energy range of
s
=
92
GeV–3 TeV is presented in view of recent and expected LHC results, experiments from low-energy as well ...as astroparticle physics. The report focusses in particular on Higgs-boson, top-quark and electroweak precision physics, but also discusses several models of beyond the standard model physics such as supersymmetry, little Higgs models and extra gauge bosons. The connection to cosmology has been analysed as well.
We report on the measurements of charge transfer in Gas Electron Multipliers (GEM) structures in high magnetic fields. These were performed in the framework of the R&D work for a Time Projection ...Chamber at a future Linear Collider. A small test chamber has been installed in the aperture of a superconducting magnet with the GEM structures mounted perpendicular to the B-field direction. The charge transfer is derived from the electrical currents monitored during irradiation with an 55Fe source. No significant loss of primary ionisation charge is observed, and an improved ion feedback suppression is achieved for high magnetic fields. Additionally, the width of the charge cloud released by individual 55Fe photons is measured using a finely segmented strip readout after the triple GEM structure. Charge widths between 0.3 and 0.5mm RMS are observed, which originate from the charge broadening inside the GEM amplification. This charge broadening is only partly suppressed at high magnetic fields.
The Compact Muon Solenoid (CMS) is one of two general purpose detectors which are foreseen to operate at the Large Hadron Collider (LHC), which is presently being built at the European laboratory for ...particle physics (CERN) in Switzerland. The Central Tracker of CMS consists of a Pixel System, which is located close to the interaction point and a Silicon Strip Tracker (SST) which instruments the intermediate and outer region. The SST is composed of 15148 Silicon Microstrip Detector Modules which contain the readout electronics (hybrids) and sensors. These modules will be assembled into substructures with control electronics and optics for transmitting data. The substructures will be integrated into the subsystems of the SST. The SST will be operated for up to ten years in the harsh radiation environment of the LHC. The lifetime of the SST will be extended by operating the detector at lowered temperature. The sensors, which are very delicate with respect to radiation damage, will be operated at a maximum temperature of -10degC. Since the assembly of the modules as well as the mounting on substructures is done at room temperature, tests in a CMS-like environment are necessary to prove the mechanical and electrical stability
The determination of the centre-of-mass energies from the LEP1 data for 1993, 1994 and 1995 is presented. Accurate knowledge of these energies is crucial in the measurement of the Z resonance ...parameters. The improved understanding of the LEP energy behaviour accumulated during the 1995 energy scan is detailed, while the 1993 and 1994 measurements are revised. For 1993 these supersede the previously published values. Additional instrumentation has allowed the detection of an unexpectedly large energy rise during physics fills. This new effect is accommodated in the modelling of the beam-energy in 1995 and propagated to the 1993 and 1994 energies. New results are reported on the magnet temperature behaviour which constitutes one of the major corrections to the average LEP energy. The 1995 energy scan took place in conditions very different from the previous years. In particular the interaction-point specific corrections to the centre-of-mass energy in 1995 are more complicated than previously: these arise from the modified radiofrequency-system configuration and from opposite-sign vertical dispersion induced by the bunch-train mode of LEP operation. Finally an improved evaluation of the LEP centre-of-mass energy spread is presented. This significantly improves the precision on the Z width.
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