7196 research outputs found
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Tackling Challenging Targets with High-Throughput Biophysical Screening at Novartis
No full textThis application note describes how Novartis successfully identified novel drug candidates
for challenging protein targets by pioneering high-throughput SPR screening. To accomplish
this, they performed their screens on a Sierra SPR®-32† instrument and analyzed their data
using Genedata Screener®. This enabled them to:
• Scale up biophysical screening and employ a multiplexed assay design for initial hit-finding
campaigns. This endeavor allowed for primary screening against mutant forms of the
protein target, resulting in the discovery of novel chemical scaffolds.
• Rapidly process complete campaigns by automating sensorgram processing and
calculations, while giving scientists full control over result review and ability to adapt the
analysis to their workflows.
• Unify data analysis into a single software platform, increasing efficiency and data
traceability
People of TM: Video of Courtney Horvath
No full textThe video will be used for an external social media engagement campaign on platforms like linked-in, facebook etc. featuring stories of people in TM.
No IP related content
Degenerative joint disease induced by repeated intra-articular injections of monosodium urate crystals in rats as investigated by translational imaging
No full textThe objective of this work was to assess the consequences of repeated intra-articular injection of monosodium urate (MSU) crystals with inflammasome priming by lipopolysaccharide (LPS) in order to simulate recurrent bouts of gout in rats. Translational imaging was applied to simultaneously detect and quantify injury in different areas of the knee joint. MSU/LPS induced joint swelling, synovial membrane thickening, fibrosis of the infrapatellar fat pad, tidemark breaching, and cartilage invasion by inflammatory cells. A higher sensitivity to mechanical stimulus was detected in paws of limbs receiving MSU/LPS compared to saline-injected limbs. In MSU/LPS-challenged joints, magnetic resonance imaging (MRI) revealed increased synovial fluid volume in the posterior region of the joint, alterations in the infrapatellar fat pad reflecting a progressive decrease of fat volume and fibrosis formation, and a significant increase in the relaxation time T2 in femoral cartilage, consistent with a reduction of proteoglycan content. MRI also showed cyst formation in the tibia, femur remodeling, and T2 reductions in extensor muscles consistent with fibrosis development. Repeated intra-articular MSU/LPS injections in the rat knee joint induced pathology in multiple tissues and may be a useful means to investigate the relationship between urate crystal deposition and the development of degenerative joint disease
Sustained Trem2 stabilization accelerates microglia heterogeneity and Aβ pathology in a mouse model of Alzheimer’s disease
No full textTREM2 is a transmembrane protein expressed exclusively in microglia in the brain that regulates inflammatory responses to pathological conditions. Proteolytic cleavage of membrane TREM2 affects microglial function and is associated with Alzheimer’s disease, but the consequence of reduced TREM2 proteolytic cleavage has not been determined. Here, we generate a transgenic mouse model of reduced Trem2 shedding (Trem2-Ile-Pro-Asp [IPD]) through amino-acid substitution of an ADAM-protease recognition site. We show that Trem2-IPD mice display increased Trem2 cell-surface-receptor load, survival, and function in myeloid cells. Using single-cell transcriptomic profiling of mouse cortex, we show that sustained Trem2 stabilization induces a shift of fate in microglial maturation and accelerates microglial responses to Aβ pathology in a mouse model of Alzheimer’s disease. Our data indicate that reduction of Trem2 proteolytic cleavage aggravates neuroinflammation during the course of Alzheimer’s disease pathology, suggesting that TREM2 shedding is a critical regulator of microglial activity in pathological states
Inhibiting IL-1beta-mediated inflammation in pediatric and young adult patients with sickle cell anemia: a randomized, placebo-controlled, double-blind trial with canakinumab (ACZ885)
No full textExcessive intravascular release of lysed cellular contents from damaged red blood cells (RBCs) in patients with sickle cell anemia (SCA) can activate the inflammasome, a multiprotein oligomer promoting maturation and secretion of pro-inflammatory cytokines, including interleukin 1-beta (IL-1). We hypothesized that IL-1 blockade by canakinumab in patients with SCA would reduce markers of inflammation and clinical disease activity. In this randomized, double-blind, multi-center phase 2a study, patients aged 8-20 years old with SCA (HbSS or HbS0thalassemia), history of acute pain episodes and elevated hsCRP >1.0 mg/L at screening were randomized 1:1 to received 6 monthly treatments with 300 mg s.c. canakinumab or placebo. Measured outcomes at baseline and weeks 4, 8, 12, 16, 20 and 24 included electronic patient-reported outcomes, hospitalization rate and adverse events (AEs) and serious AEs (SAEs). All but one of the 49 enrolled patients received background hydroxyurea therapy. Although the primary objective (pre-specified reduction of pain) was not met, compared to placebo-arm patients, canakinumab-treated patients had reductions in markers of inflammation, incidence of SCA-related AE and SAE, and number and duration of hospitalizations, as well as trends for improvement in pain intensity, fatigue and absences from school or work. Post-hoc analysis revealed treatment effects on weight gain in, restricted to pediatric patients. Canakinumab was well tolerated with no treatment-related SAEs and no new safety finding. These findings demonstrate that the inflammation associated with SCA can be reduced by selective IL-1b blockade by canakinumab with potential for therapeutic benefits. This trial was registered at www.clinicaltrials.gov as NCT02961218
Twin-screw melt granulation based high drug load tablet formulation. An industrial perspective on process transfer to a commercial manufacturing site
No full textOur motivation for this paper is to highlight select aspects of pharmaceutical processes (granulation and tablet compression) and process analytical technology (PAT) that were performed as part of the commercial manufacturing site transfer campaign of high drug load (80.5 % w/w) immediate release QAW039/Fevipiprant tablets. Twin-screw melt granulation (TSMG) was the key unit operation to achieve a high drug load granule (91.5 % w/w), resulting in a reduced pill burden for the compositional identical 150 mg and 450 mg doses. A proven acceptable range (PAR) was established during the campaign by varying process parameters for granulation (screw speed), blend lubrication, and tabletting (dwell time and compression force) at three process levels (upper, target, lower). These combinations were selected to have the same effect on critical quality attributes (CQAs) i.e., lower (-) and upper (+) process level challenged tablet aspect and dissolution, respectively. Melt granulation was successfully performed using a 50 mm twin-screw extruder at constant feed rate (50 kg/h) across the PAR. Compression of the six final blends (~300 kg/batch) showed no impact of varied granulation and compression process conditions on both CQAs, thereby establishing the PAR within the parameters investigated for a commercially viable process transfer. This paper shows the use of PAT for process development with the goal of readiness of real time release testing (RTRT) which allowed a cost-effective control of the manufacturing process to meet all quality and product specifications. In this context, a near infrared spectroscopy (NIRS) based PAT to determine uniformity of dosage units by content uniformity, assay and identity was also developed and validated to predict CQAs on core (uncoated) tablets in preparation for RTRT of future commercial batches
Discovery, preclinical characterization, and early clinical activity of JDQ443, a structurally novel, highly potent, selective covalent oral inhibitor of KRASG12C
No full textCovalent inhibitors of KRASG12C have shown antitumor activity against advanced/metastatic KRAS G12C-mutated cancers, though resistance emerges and additional strategies are needed to improve outcomes. JDQ443 is a structurally unique covalent inhibitor of GDP-bound KRASG12C that forms novel interactions with the switch II pocket. JDQ443 potently inhibits KRASG12C-driven cellular signaling and demonstrates selective antiproliferative activity in KRAS G12C-mutated cell lines, including those with G12C/H95 double mutations. In vivo, JDQ443 induces AUC exposure-driven antitumor efficacy in KRAS G12C-mutated cell-derived (CDX) and patient-derived (PDX) tumor xenografts. In PDX models, single-agent JDQ443 activity is enhanced by combination with SHP2, MEK or CDK4/6 inhibitors. Notably, the benefit of JDQ443 plus the SHP2 inhibitor TNO155 is maintained at reduced doses of either agent in CDX models, consistent with mechanistic synergy. JDQ443 is in clinical development as monotherapy and in combination with TNO155, with both strategies showing antitumor activity in patients with KRAS G12C-mutated tumors
Substituents of life: The most common substituent patterns present in natural products.
No full textComparison of substituents present in natural products with the substituents found in average synthetic molecules reveals considerable differences between these two groups. The natural products substituents contain mostly oxygen heteroatoms, are structurally more complex, often containing double bonds and are rich in stereocenters. Substituents found in synthetic molecules contain nitrogen and sulfur heteroatoms, halogenes and more aromatic and particularly heteroaromatic rings. The characteristics of substituents typical for natural products identified here can be useful in the medicinal chemistry context, for example to guide the synthesis of natural product-like libraries and natural product-inspired fragment collections. The results may be used also to support compound derivatization strategies and the design of pseudo-natural natural products
3R measures in facilities for the production of genetically modified rodents.
No full textSociocultural changes in the human-animal relationship have led to increasing demands for animal welfare in biomedical research. The 3R concept is the basis for bringing this demand into practice: Replace animal experiments with alternatives where possible, Reduce the number of animals used to a scientifically justified minimum and Refine the procedure to minimize animal harm. The generation of gene-modified sentient animals such as mice and rats involves many steps that include various forms of manipulation. So far, no coherent analysis of the application of the 3Rs to gene manipulation has been performed. Here we provide guidelines from the Committee on Genetics and Breeding of Laboratory Animals of the German Society for Laboratory Animal Science to implement the 3Rs in every step during the generation of genetically modified animals. We provide recommendations for applying the 3Rs as well as success/intervention parameters for each step of the process, from experiment planning to choice of technology, harm-benefit analysis, husbandry conditions, management of genetically modified lines and actual procedures. We also discuss future challenges for animal welfare in the context of developing technologies. Taken together, we expect that our comprehensive analysis and our recommendations for the appropriate implementation of the 3Rs to technologies for genetic modifications of rodents will benefit scientists from a wide range of disciplines and will help to improve the welfare of a large number of laboratory animals worldwide
IL-17-induced dimerization of IL-17RA drives the formation of the IL-17 signalosome to potentiate signaling.
No full textSignaling through innate immune receptors such as the Toll-like receptor (TLR)/interleukin-1 receptor (IL-1R) superfamily proceeds via the assembly of large membrane-proximal complexes or "signalosomes." Although structurally distinct, the IL-17 receptor family triggers cellular responses that are typical of innate immune receptors. The IL-17RA receptor subunit is shared by several members of the IL-17 family. Using a combination of crystallographic, biophysical, and mutational studies, we show that IL-17A, IL-17F, and IL-17A/F induce IL-17RA dimerization. X-ray analysis of the heteromeric IL-17A complex with the extracellular domains of the IL-17RA and IL-17RC receptors reveals that cytokine-induced IL-17RA dimerization leads to the formation of a 2:2:2 hexameric signaling assembly. Furthermore, we demonstrate that the formation of the IL-17 signalosome potentiates IL-17-induced IL-36γ and CXCL1 mRNA expression in human keratinocytes, compared with a dimerization-defective IL-17RA variant