scholarly journals Targeting the water network in cyclin G associated kinase (GAK) with 4-anilino-quin(az)oline inhibitors

2020 ◽  
Author(s):  
Christopher R. M. Asquith ◽  
Graham J. Tizzard ◽  
James M. Bennett ◽  
Carrow I. Wells ◽  
Jonathan M. Elkins ◽  
...  
Keyword(s):  
Kinase Inhibitor ◽  
Great Promise ◽  
Target Engagement ◽  
Inhibitor Design ◽  
Water Network ◽  
Atp Binding Site ◽  
Cellular Target ◽  
Cyclin G ◽  
Design And Testing ◽  
Targeted Library

AbstractWater networks within kinase inhibitor design and more widely within drug discovery are generally poorly understood. The successful targeting of these networks prospectively has great promise for all facets of inhibitor design, including potency and selectivity on target. Here we describe the design and testing of a targeted library of 4-anilinoquinolines for use as inhibitors of cyclin G associated kinase (GAK). The GAK cellular target engagement assays, ATP binding site modelling and extensive water mapping provide a clear route to access potent inhibitors for GAK and beyond.

Rapid Evaluation of Small Molecule Cellular Target Engagement with a Luminescent Thermal Shift Assay

2021 ◽  
Author(s):  
Jonathan D. Mortison ◽  
Ivan Cornella-Taracido ◽  
Gireedhar Venkatchalam ◽  
Anthony W. Partridge ◽  
Nirodhini Siriwardana ◽  
...  
Keyword(s):  
Small Molecule ◽  
Rapid Evaluation ◽  
Target Engagement ◽  
Thermal Shift Assay ◽  
Cellular Target ◽  
Thermal Shift

scholarly journals Evaluation of the binding affinity of E3 ubiquitin ligase ligands by cellular target engagement and in-cell ELISA assay

2021 ◽  
Vol 2 (1) ◽  
pp. 100288
Author(s):  
Ka Yang ◽  
Yaxian Zhou ◽  
Brett L. Roberts ◽  
Xueqing Nie ◽  
Weiping Tang
Keyword(s):  
Binding Affinity ◽  
Ubiquitin Ligase ◽  
E3 Ubiquitin Ligase ◽  
Elisa Assay ◽  
Target Engagement ◽  
Cellular Target ◽  
Cell Elisa

Characterization of an Aromatic Trifluoromethyl Ketone as a New Warhead for Covalently Reversible Kinase Inhibitor Design

2021 ◽  
pp. 116457
Author(s):  
Zhen Zhang ◽  
Yongjin Wang ◽  
Xiaojuan Chen ◽  
Xiaojuan Song ◽  
Zhengchao Tu ◽  
...  
Keyword(s):  
Kinase Inhibitor ◽  
Inhibitor Design

scholarly journals A cellular target engagement assay for the characterization of SHP2 (PTPN11) phosphatase inhibitors

2020 ◽  
Vol 295 (9) ◽  
pp. 2601-2613 ◽  
Author(s):  
Celeste Romero ◽  
Lester J. Lambert ◽  
Douglas J. Sheffler ◽  
Laurent J. S. De Backer ◽  
Dhanya Raveendra-Panickar ◽  
...  
Keyword(s):  
Developmental Disorders ◽  
Tyrosine Phosphatase ◽  
Therapeutic Window ◽  
Target Engagement ◽  
Allosteric Inhibitors ◽  
Intact Cells ◽  
Phosphatase Inhibitors ◽  
Increased Risk ◽  
Cellular Target ◽  
Risk Of Malignancy

The nonreceptor protein-tyrosine phosphatase (PTP) SHP2 is encoded by the proto-oncogene PTPN11 and is a ubiquitously expressed key regulator of cell signaling, acting on a number of cellular processes and components, including the Ras/Raf/Erk, PI3K/Akt, and JAK/STAT pathways and immune checkpoint receptors. Aberrant SHP2 activity has been implicated in all phases of tumor initiation, progression, and metastasis. Gain-of-function PTPN11 mutations drive oncogenesis in several leukemias and cause developmental disorders with increased risk of malignancy such as Noonan syndrome. Until recently, small molecule-based targeting of SHP2 was hampered by the failure of orthosteric active-site inhibitors to achieve selectivity and potency within a useful therapeutic window. However, new SHP2 allosteric inhibitors with excellent potency and selectivity have sparked renewed interest in the selective targeting of SHP2 and other PTP family members. Crucially, drug discovery campaigns focusing on SHP2 would greatly benefit from the ability to validate the cellular target engagement of candidate inhibitors. Here, we report a cellular thermal shift assay that reliably detects target engagement of SHP2 inhibitors. Using this assay, based on the DiscoverX InCell Pulse enzyme complementation technology, we characterized the binding of several SHP2 allosteric inhibitors in intact cells. Moreover, we demonstrate the robustness and reliability of a 384-well miniaturized version of the assay for the screening of SHP2 inhibitors targeting either WT SHP2 or its oncogenic E76K variant. Finally, we provide an example of the assay's ability to identify and characterize novel compounds with specific cellular potency for either WT or mutant SHP2.

Cellular target engagement: a new paradigm in drug discovery

2018 ◽  
Vol 10 (14) ◽  
pp. 1641-1644 ◽  
Author(s):  
Ivan Babic ◽  
Santosh Kesari ◽  
Elmar Nurmemmedov
Keyword(s):  
Drug Discovery ◽  
Target Engagement ◽  
New Paradigm ◽  
Cellular Target

scholarly journals Discovery of an MLLT1/3 YEATS Domain Chemical Probe

2018 ◽  
Author(s):  
Moses Moustakim ◽  
Thomas Christott ◽  
Octovia P. Monteiro ◽  
James Bennett ◽  
Charline Giroud ◽  
...  
Keyword(s):  
Drug Discovery ◽  
Crystal Structures ◽  
Small Molecule ◽  
Therapeutic Potential ◽  
Selective Inhibitor ◽  
Target Engagement ◽  
Chemical Probe ◽  
X Ray ◽  
Cellular Target ◽  
Excellent Selectivity

<p>YEATS domain (YD) containing proteins are an emerging</p> <p>class of epigenetic targets in drug discovery. Dysregulation of these modified lysine binding proteins has been linked to the onset and progression of cancers. We herein report the discovery and characterisation of the first small molecule chemical probe, SGC-iMLLT, for the YD of MLLT1 (ENL/YEATS1) and MLLT3 (AF9/YEATS3). SGC-iMLLT is a potent and selective inhibitor of MLLT1/3 -histone interactions. Excellent selectivity over other human YD proteins (YEATS2/4) and bromodomains was observed. Furthermore, our probe displays cellular target engagement of MLLT1 and MLLT3. The first small molecule X-ray co-crystal structures with the MLLT1 YD are also reported. This first in class probe molecule can be used to understand MLLT1/3 associated biology and the therapeutic potential of small molecule YD inhibitors.</p>

scholarly journals ABPP-HT - high-throughput activity-based profiling of deubiquitylating enzyme inhibitors in a cellular context

2020 ◽  
Author(s):  
Hannah Jones ◽  
Raphael Heilig ◽  
Roman Fischer ◽  
Benedikt M Kessler ◽  
Adan Pinto-Fernandez
Keyword(s):  
Sample Preparation ◽  
High Throughput ◽  
Enzyme Inhibitors ◽  
Drug Efficacy ◽  
Protein Profiling ◽  
Target Engagement ◽  
Molecule Inhibitor ◽  
Cellular Target ◽  
Cellular Context ◽  
Target Effects

AbstractThe potency and selectivity of a small molecule inhibitor are key parameters to assess during the early stages of drug discovery. In particular, it is very informative for characterizing compounds in a relevant cellular context in order to reveal potential off-target effects and drug efficacy. Activity-based probes (ABPs) are valuable tools for that purpose, however, obtaining cellular target engagement data in a high-throughput format has been particularly challenging. Here, we describe a new methodology named ABPP-HT (high-throughput-compatible activity-based protein profiling), implementing a semi-automated proteomic sample preparation workflow that increases the throughput capabilities of the classical ABPP workflow approximately ten times while preserving its enzyme profiling characteristics. Using a panel of deubiquitylating enzyme (DUB) inhibitors, we demonstrate the feasibility of ABPP-HT to provide compound selectivity profiles of endogenous DUBs in a cellular context at a fraction of time as compared to previous methodologies.

scholarly journals Positioning High-Throughput CETSA in Early Drug Discovery through Screening against B-Raf and PARP1

2018 ◽  
Vol 24 (2) ◽  
pp. 121-132 ◽  
Author(s):  
Joseph Shaw ◽  
Ian Dale ◽  
Paul Hemsley ◽  
Lindsey Leach ◽  
Nancy Dekki ◽  
...  
Keyword(s):  
Drug Discovery ◽  
High Throughput ◽  
Target Engagement ◽  
Native State ◽  
Cellular Target ◽  
Protein Thermal Stability ◽  
Cellular Thermal Shift Assay ◽  
Hit Identification ◽  
Early Drug ◽  
Thermal Shift

Methods to measure cellular target engagement are increasingly being used in early drug discovery. The Cellular Thermal Shift Assay (CETSA) is one such method. CETSA can investigate target engagement by measuring changes in protein thermal stability upon compound binding within the intracellular environment. It can be performed in high-throughput, microplate-based formats to enable broader application to early drug discovery campaigns, though high-throughput forms of CETSA have only been reported for a limited number of targets. CETSA offers the advantage of investigating the target of interest in its physiological environment and native state, but it is not clear yet how well this technology correlates to more established and conventional cellular and biochemical approaches widely used in drug discovery. We report two novel high-throughput CETSA (CETSA HT) assays for B-Raf and PARP1, demonstrating the application of this technology to additional targets. By performing comparative analyses with other assays, we show that CETSA HT correlates well with other screening technologies and can be applied throughout various stages of hit identification and lead optimization. Our results support the use of CETSA HT as a broadly applicable and valuable methodology to help drive drug discovery campaigns to molecules that engage the intended target in cells.

scholarly journals A new “angle” on kinase inhibitor design: Prioritizing amphosteric activity above kinase inhibition

2015 ◽  
Vol 2 (2) ◽  
pp. e975641 ◽  
Author(s):  
Justin G Meyerowitz ◽  
William A Weiss ◽  
W Clay Gustafson
Keyword(s):  
Kinase Inhibitor ◽  
Inhibitor Design ◽  
Kinase Inhibition
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