January 12-16, 2013
2013 SLAS - 2nd Annual Conference & Exhibition
Gaylord Palms Resort and Convention Center; Orlando, Florida
DiscoveRx is a proud sponsor of of the second annual SLAS2013 (Society for Laboratory Automation and Screening) Conference & Exhibition, located in Orlando Florida at the Gaylord Palms Resort and Convention Center, January 12-16, 2013. The SLAS2013 Exhibition allows participants to make important connections and find repeatable inspiration in the innovative technology products and services that are showcased. Go to the SLAS website »
SLAS Satellite Symposium at the Sanford-Burnham Medical Research Institute in Lake Nona
Sunday, January 13th, 2013; 1:00PM until 5:00PM
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BioMAP® Systems A Phenotypic Assay Platform for Reducing Preclinical Attrition
Tuesday, Jan. 15 – 8:00 – 8:45 AM
Location: Naples Room 2-3
The BioMAP® systems platform, utilizing primary human cell-based tissue and disease models, provides a preview of in vivo drug effects. Case studies will be presented that...
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Targeting the Epigenome: Unexpected compound promiscuity and target class cross-reactivity revealed using the industry’s largest bromodomain profiling and cell-based assay platforms
Tuesday, Jan. 15 – 2:00 – 2:45 PM
Location: Sanibel Room 1-3
Bromodomain containing proteins are a newly established class of druggable targets implicated as major hubs in oncology and inflammation. Here we describe inhibitor specificity...
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Monday, January 14, 2013 1:00PM – 3:00PM
A Novel Assay platform for the Detection of Kinase-Inhibitor Binding in Intact Mammalian Cells (MP 147)
Kinase binding assays have become an integral part of inhibitor characterization. These tools have been applied in KINOME-wide screening systems as well as the detailed characterization of inhibitor residence time for lead optimization and MOA studies. However in cells, assays are limited to monitoring kinase activity and downstream substrate phosphorylation events. Although these types of cellular assays are critical to move programs forward, their ability to characterize inhibitor function is limited, requires targeted antibody reagents, and knowledge of specific substrates which are often not available. To circumvent these issues and apply the power of binding assays to a cellular format we have devised a method for monitoring compound binding to kinase targets in intact mammalian cells termed InCELL Hunter™. The assay is shown to retain the benefits of cellular assays such as assessing compound permeability, toxicity, and target engagement; while exhibiting the generic applicability of a kinase binding assay. The InCELL Hunter™ assay correctly describes type I, type II, and Type III inhibitors with the expected rank order cellular potencies. The assay is activity and substrate independent making it amenable to previously intractable targets and provides novel cellular information that complements both biochemical and existing cellular formats.
BROMOscanSM – A High Throughput, Quantitative Ligand Binding Platform Identifies Best-In-Class Bromodomain Inhibitors From a Screen of Mature Compounds Targeting Other Protein Classes (MP 148)
Post-translationally appended acetyllysine marks on histone tails are key regulatory features of the epigenetic code. Bromodomains are “readers” of this specific lysine acetylation code, playing an important role in chromatin remodeling and regulation of gene expression. Bromodomains have emerged as an important new druggable target class in small-molecule inhibitor drug discovery, and several bromodomain-containing proteins have been associated with disease. There are 57 bromodomains contained in 41 different proteins; however, few small molecule bromodomain inhibitors have been reported. One primary factor limiting the discovery of new inhibitors is the absence of a comprehensive biochemical bromodomain screening platform. Here we describe the application of DiscoveRx Corporation’s proven ligand binding assay technology (KINOMEscan) to the development of quantitative ligand binding assays for human bromodomains (BROMOscan). We have developed a carefully validated assay panel that covers >30 percent of the human bromodomain family, and this panel is suitable for HTS, selectivity profiling, and quantitative affinity (Kd) assessment. We have used this panel internally to identify novel bromodomain inhibitors and, remarkably, have demonstrated that known, mature inhibitors thought to be selective for targets from other protein families have best in class affinity for bromodomains as well. These data shall be presented, as will a description of the BROMOscan panel replete with extensive assay validation data.
InCELL Hunter™: A novel cellular assay platform for the direct measurement of compound binding to intracellular targets in intact mammalian cells (MP 152)
Assays that directly detect compound binding can be of great utility in translating in vitro data to more biologically relevant models. However for intracellular targets, these types of binding assays in cells have not been possible. Cellular assays for these targets are limited to downstream activity-based read-outs which can be difficult to develop, may not be proximal to the target, or may require reagents or knowledge that does not currently exist. Here we describe the development of a novel cell-based assay for the detection of compound binding to intracellular proteins termed InCELL Hunter™. Using this assay, knowledge of downstream substrates, or catalytic activity of the target is not necessary for signal generation relieving many of the constraints of current cell-based assay technologies. We show the generic application of this technology to a number of challenging targets including methyltransferases, bromodomains, and kinases. The assay is shown to report cellular permeability and appropriately report compound activity making this a promising platform for bridging the gap between in vitro and cellular assays.
scanKINETIC - A Broadly Applicable Tool for Kinetic Characterization of Interactions Between Small Molecule Inhibitors and Protein Targets From the Kinase and Bromodomain Families (TP 181)
Inhibitor association and dissociation kinetics can have important effects on a compound’s drug-like properties and in vitro, cellular, and in vivo behaviors. The association and dissociation rates for protein target-inhibitor complexes are variable and can be both inhibitor and protein-dependent. In addition, it is often important to classify inhibitors as being reversible or irreversible (covalent). While many inhibitor complexes form and dissociate rapidly (< minutes), others can have extraordinarily low association and/or dissociation rates, requiring several hours to reach equilibrium. Slowly associating and/or dissociating inhibitors can yield conflicting potency data in various in vitro, cellular, and in vivo settings, where the inhibitor-target equilibration time is often assay specific. Though there are several methods available for measuring inhibitor binding kinetics, most available offerings currently address only a small subset of relevant protein targets. Here we describe scanKINETIC, a semi-quantitative, broadly applicable tool based on proven KINOMEscan technology, which enables the kinetic characterization of inhibitor interactions. scanKINETIC is currently applicable to a majority of the 518 known protein kinases and for a rich sampling of lipid, atypical, and clinically relevant mutant kinases. In addition, scanKINETIC can currently be applied to greater than 30 percent of bromodomain epigenetic reader proteins. Relevant exemplary data for known inhibitors will be presented and discussed.
Multi-parameter GPCR Screening: An essential tool for Uncovering GPCR Mutant phenotypes (TP 182)
The recent appreciation of functional selectivity/ligand bias in GPCR signaling has uncovered new opportunities for therapeutic discovery and continues to increase our understanding of compound function. We reasoned that since ligands and hence receptor conformation can differentially stimulate particular signaling pathways, variations in amino acid composition could similarly bias receptors and their responses to ligand. To test this hypothesis, we have compared a number of common GPCR variants and their wild-type counterparts in their ability to traffic to the cell surface and signal through g-proteins, arrestin, and internalization. This comprehensive suite of assays appropriately described mutants that caused GPCR misfolding and ER retention, as well as detailed changes in the functional selectivity of compounds. Thus the combination of these platforms provides an unprecedented view of GPCR variant activity in a quantifiable and comparable manner amenable to paneling large numbers of variants. These tools can be used identify new prospects for existing compounds and stratify patient populations for similar responses to drug candidates.