3 research outputs found
A signalome screening approach in the autoinflammatory disease TNF Receptor Associated Periodic Syndrome (TRAPS) highlights the anti-inflammatory properties of drugs for repurposing
TNF Receptor Associated Periodic Syndrome (TRAPS) is an autoinflammatory disease caused by mutations in TNF Receptor 1 (TNFR1). Current therapies for TRAPS are limited and do not target the pro-inflammatory signalling pathways that are central to the disease mechanism. Our aim was to identify drugs for repurposing as anti-inflammatories based on their ability to down-regulate molecules associated with inflammatory signalling pathways that are activated in TRAPS. This was achieved using rigorously optimised, high through- put cell culture and reverse phase protein microarray systems to screen compounds for their effects on the TRAPS-associated inflammatory signalome. 1360 approved, publically available, pharmacologically active substances were investigated for their effects on 40 signalling molecules associated with pro-inflammatory signalling pathways that are constitutively upregulated in TRAPS. The drugs were screened at four ten-fold concentrations on cell lines expressing both wild-type (WT) TNFR1 and TRAPS-associated C33Y mutant TNFR1, or WT TNFR1 alone; signalling molecule levels were then determined in cell lysates by the reverse phase protein microarray. A novel mathematical methodology was developed to rank the compounds for their ability to reduce the expression of signalling molecules in the C33Y-TNFR1 transfectants towards the level seen in the WT-TNFR1 transfectants. Seven high-ranking drugs were selected and tested by RPPA for effects on the same 40 signalling molecules in lysates of peripheral blood mononuclear cells (PBMCs) from C33Y-TRAPS patients compared to PBMCs from normal controls. The fluoroquinolone antibiotic lomefloxacin, as well as others from this class of compounds, showed the most significant effects on multiple pro-inflammatory signalling pathways that are constitutively activated in TRAPS; lomefloxacin dose-dependently significantly reduced expression of 7/40 signalling molecules across the Jak/Stat, MAPK, NF-kB and PI3K/AKT pathways. This study demonstrates the power of signalome screening for identifying candidates for drug repurposing
Structure-Guided Discovery of Potent and Selective Inhibitors of ERK1/2 from a Modestly Active and Promiscuous Chemical Start Point
There are a number of small-molecule
inhibitors targeting the RAS/RAF/MEK/ERK
signaling pathway that have either been approved or are in clinical
development for oncology across a range of disease indications. The
inhibition of ERK1/2 is of significant current interest, as cell lines
with acquired resistance to BRAF and MEK inhibitors have been shown
to maintain sensitivity to ERK1/2 inhibition in preclinical models.
This article reports on our recent work to identify novel, potent,
and selective reversible ERK1/2 inhibitors from a low-molecular-weight,
modestly active, and highly promiscuous chemical start point, compound <b>4</b>. To guide and inform the evolution of this series, inhibitor
binding mode information from X-ray crystal structures was critical
in the rapid exploration of this template to compound <b>35</b>, which was active when tested in in vivo antitumor efficacy experiments
Structure-Based Design of Potent and Selective Inhibitors of the Metabolic Kinase PFKFB3
A weak
screening hit with suboptimal physicochemical properties was optimized
against PFKFB3 kinase using critical structure-guided insights. The
resulting compounds demonstrated high selectivity over related PFKFB
isoforms and modulation of the target in a cellular context. A selected
example demonstrated exposure in animals following oral dosing. Examples
from this series may serve as useful probes to understand the emerging
biology of this metabolic target