A5054167659- Click Chemistry and Applications
- Protein Degradation and Inhibitors
- DNA Repair Mechanisms
- CRISPR and Genetic Engineering
- Chemical Synthesis and Analysis
- Renal and related cancers
- Animal Genetics and Reproduction
- Ubiquitin and proteasome pathways
- Peptidase Inhibition and Analysis
- Computational Drug Discovery Methods
- Innovative Microfluidic and Catalytic Techniques Innovation
- Synthesis and Catalytic Reactions
- Multiple Myeloma Research and Treatments
- Digestive system and related health
- Advanced Breast Cancer Therapies
- PARP inhibition in cancer therapy
- Cancer-related Molecular Pathways
- Nuclear Structure and Function
- Cancer-related gene regulation
- RNA modifications and cancer
- Biomarkers in Disease Mechanisms
- Spectroscopy Techniques in Biomedical and Chemical Research
- Protein Kinase Regulation and GTPase Signaling
- Histone Deacetylase Inhibitors Research
- Cannabis and Cannabinoid Research
GlaxoSmithKline (United Kingdom)
2019-2024
Indiana University Bloomington
2024
Age UK
2024
University of Strathclyde
2019-2023
AstraZeneca (Brazil)
2022
University of Sheffield
2017-2022
Abstract Micronuclei represent the cellular attempt to compartmentalize DNA maintain genomic integrity threatened by mitotic errors and genotoxic events. Some micronuclei show aberrant nuclear envelopes (NEs) that collapse, generating damaged can promote complex genome alterations. However, ruptured also provide a pool of cytosolic stimulate antitumor immunity, revealing complexity micronuclear impact on tumor progression. The ESCRT-III (Endosomal Sorting Complex Required for Transport-III)...
Here, we report a comprehensive profiling of sulfur(VI) fluorides (SVI-Fs) as reactive groups for chemical biology applications. SVI-Fs are functionalities that modify lysine, tyrosine, histidine, and serine sidechains. A panel were studied with respect to hydrolytic stability reactivity nucleophilic amino acid The use covalently carbonic anhydrase II (CAII) range kinases was then investigated. Finally, the SVI-F used in live cell chemoproteomic workflows, identifying novel protein targets...
Carboxylesterases (CEs) are a class of enzymes that catalyze the hydrolysis esters in variety endogenous and exogenous molecules. CEs play an important role drug metabolism, onset progression disease, can be harnessed for prodrug activation strategies. As such, regulation is clinical pharmaceutical consideration. Here, we report first ratiometric sensor CE activity using Raman spectroscopy based on bisarylbutadiyne scaffold. The was shown to highly sensitive specific detection had low...
Microsatellite-unstable (MSI) cancers require WRN helicase to resolve replication stress due expanded DNA (TA)n dinucleotide repeats. is a promising synthetic lethal target for MSI tumors, and inhibitors are in development. In this study, we used CRISPR-Cas9 base editing map residues critical cells, validating the domain as primary drug target. Fragment-based screening led development of potent highly selective covalent inhibitors. These compounds selectively suppressed model growth vitro...
Poly(ADP-ribosylation) of proteins following DNA damage is well studied and the use poly(ADP-ribose) polymerase (PARP) inhibitors as therapeutic agents an exciting prospect for treatment many cancers. Poly(ADP-ribose) glycohydrolase (PARG) has endo- exoglycosidase activities which can cleave glycosidic bonds, rapidly reversing action PARP enzymes. Like addition (PAR) by PARP, removal PAR PARG also thought to be required repair strand breaks continued replication at perturbed forks. Here we...
Methods for rapid identification of chemical tools are essential the validation emerging targets and to provide medicinal chemistry starting points development new medicines. Here, we report a screening platform that combines 'direct-to-biology' high-throughput (D2B-HTC) with photoreactive fragments. The enabled synthesis >1000 PhotoAffinity Bits (HTC-PhABits) in 384-well plates 24 h their subsequent as crude reaction products protein target without purification. Screening HTC-PhABit library...
Upon DNA binding the poly(ADP-ribose) polymerase family of enzymes (PARPs) add multiple ADP-ribose subunits to themselves and other acceptor proteins. Inhibitors PARPs have become an exciting real prospect for monotherapy as sensitizers ionising radiation (IR). The action are reversed by glycohydrolase (PARG). Until recently studies PARG been limited lack inhibitor. Here, a first in class, specific, cell permeable inhibitor, PDD00017273, is shown radiosensitize. Further, PDD00017273 compared...
Advances in genomic analyses enable the identification of new proteins that are associated with disease. To validate these targets, tool molecules required to demonstrate a ligand can have disease-modifying effect. Currently, as tools reported for only fraction proteome, platforms discovery essential leverage insights from analyses. Fragment screening offers an efficient approach explore chemical space. Presented here is fragment-screening platform, termed PhABits (PhotoAffinity Bits), which...
Abstract The CDK family plays a crucial role in the control of cell cycle. Dysregulation and mutation CDKs has been implicated cancer have investigated extensively as potential therapeutic targets. Selective inhibition specific isoforms is to achieve effect while minimising toxicity. We present group photoaffinity probes designed bind CDKs. site crosslinking optimised probe, well its ability enrich members from lysates, was investigated. In proof concept study, we subsequently developed...
We report a ‘direct-to-biology’ (D2B) approach for optimising covalent acrylamide binders of protein targets and apply this to the identification selective cell-active inhibitor Werner (WRN) helicase. Inhibition WRN helicase activity exhibits synthetic lethal relationship with cancers displaying high microsatellite instability (MSI-H) is being pursued as therapeutic strategy in clinic. Using intact-protein liquid chromatography-mass spectrometry (LC-MS) screening, we identified fragment...
We present a carboxylate-targeting reactive fragment screening platform using 2-aryl-5-carboxytetrazole (ACT) as the photoreactive functionality. This work will provide simple accessible method to rapidly discover tool molecules interrogate important biological targets.
Reactive fragment (RF) screening has emerged as an efficient method for ligand discovery across the proteome, irrespective of a target’s perceived tractability. To date, however, efficiency subsequent optimisation campaigns largely been low‐throughput, constrained by need synthesis and purification target compounds. We report high‐throughput platform ‘direct‐to‐biology’ (D2B) cysteine‐targeting chloroacetamide RFs, wherein is performed in 384‐well plates allowing direct assessment downstream...
Fragment screening is a popular strategy of generating viable chemical starting points especially for challenging targets. Although fragments provide better coverage space and they have typically higher chance binding, their weak affinity necessitates highly sensitive biophysical assays. Here, we introduce concept that combines evolutionary optimized fragment pharmacophores with the use photoaffinity handle enables high hit rates by LC-MS-based detection. The sensitivity our protocol was...
Chemical probes are valuable tools to explore the function of proteins. Incorporation electrophiles into small molecules enables covalent capture protein interactions and provides access powerful technologies including chemoproteomic profiling reactive fragment screening. Current approaches have been largely limited pockets containing cysteine, so establishing strategies target other amino acid residues is essential expanding applicability across proteome. Here, we profiled sulfur(VI)...
Chemoresistance poses a great barrier to breast cancer treatment and is thought correlate with increased matrix stiffness. We developed two-dimensional (2D) polyacrylamide (PAA) three-dimensional (3D) alginate in vitro models of tissue stiffness that mimic the normal cancer. then used these compare cell viability response chemotherapeutic treatment. In both 2D 3D we observed growth size was at higher corresponding tumours compared tissue. When measured, specific differential for gemcitabine...
Abstract Advances in genomic analyses enable the identification of new proteins that are associated with disease. To validate these targets, tool molecules required to demonstrate a ligand can have disease‐modifying effect. Currently, as tools reported for only fraction proteome, platforms discovery essential leverage insights from analyses. Fragment screening offers an efficient approach explore chemical space. Presented here is fragment‐screening platform, termed PhABits (PhotoAffinity...
<p>Methods for rapid identification of chemical tools are essential the validation emerging targets and to provide medicinal chemistry starting points development <a>new medicines. Here, we report a screening platform that combines ‘direct-to-biology’ high-throughput (D2B-HTC) with photoreactive covalent fragments. The enabled synthesis >1000 PhotoAffinity Bits (HTC-PhABits) in 384-well plates. Screening HTC-PhABit library </a>carbonic anhydrase I (CAI) afforded 7 hits...
Abstract The CDK family plays a crucial role in the control of cell cycle. Dysregulation and mutation CDKs has been implicated cancer have investigated extensively as potential therapeutic targets. Selective inhibition specific isoforms is to achieve effect while minimising toxicity. We present group photoaffinity probes designed bind CDKs. site crosslinking optimised probe, well its ability enrich members from lysates, was investigated. In proof concept study, we subsequently developed...
Target validation remains a challenge in drug discovery, which leads to high attrition rate the discovery process, particularly Phase II clinical trials. Consequently, new approaches enhance target are valuable tools improve process. Here we report combination of site-directed mutagenesis and electrophilic fragments enable rapid identification small molecules that selectively inhibit mutant protein. Using bromodomain- containing protein BRD4 as an example, employed structure-based approach...
The synthesis and characterisation of fluorosulfate covalent inhibitors the lipid kinase PI4KIIIβ is described. conserved lysine residue located within ATP binding site was targeted, optimised compounds based upon reversible with good activity physicochemical profile showed strong interactions slow onset times for inhibition, resulting in an excellent selectivity target. X-Ray crystallography demonstrated a distal tyrosine could also be targeted using strategy. Combination this knowledge...
Target validation remains a challenge in drug discovery, which leads to high attrition rate the discovery process, particularly Phase II clinical trials. Consequently, new approaches enhance target are valuable tools improve process. Here, we report combination of site-directed mutagenesis and electrophilic fragments enable rapid identification small molecules that selectively inhibit mutant protein. Using bromodomain-containing protein BRD4 as an example, employed structure-based approach...
<p>Supplementary Figure 1. Generating base editor-expressing MSI cell lines for screening. Supplementary 2. Quality control base-editor screens in lines. 3. Discovery and characterization of novel inhibitors WRN helicase. 4. Correlation inhibitor sensitivity with genetic molecular markers models. 5. Comparative analysis DNA damage MSS cells treated inactivation. 6: Effects GSK_WRN4 on colorectal cancer xenografts.</p>
<p>This table lists the results from Base editing screen</p>
<p>This file contains details of the BE sgRNA library</p>
<p>This file contain information on the Cell lines and organoids used in this study</p>