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Immediately preceding Structure-Based
Drug Design, June 15-16
Day 2
Tuesday, June 13
7:45am Morning Coffee
Breakfast Technology Workshop
Kinase Assay Development Strategies
Dr. Roger Bossé, Technology & BD Leader, Discovery & Research
Reagents, Molecular Medicine, PerkinElmer
Choosing an appropriate strategy for Kinase assay development can be challenging, with factors such as substrate size and antibody availability influencing the selection of the platform. This presentation will examine the various approaches and techniques that are employed. In addition, we’ll introduce a novel platform for antibody-free, label-less detection of large protein substrates. |
Sponsored
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8:40 Chairperson's Opening Remarks
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Anti-Tumor Developments
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| Featured Presentation
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8:45 Oncogenic Kinase Signaling
Dr. Peter Blume-Jensen, Senior Director, Molecular Oncology,
Merck Research Laboratories |
9:30 PI-3 Kinase and PTEN Inhibitors for Cancer Therapeutics
Dr. Don Durdon, Department of Pediatrics, Aflac Cancer Center, Emory University School of Medicine; Scientific Founder, Semafore Pharmaceuticals, Inc.
We will present preclinical results on a vascular targeted pan PI-3 kinase inhibitor, SF1126, developed by Semafore Pharmaceuticals in collaboration with the Aflac Cancer Center at the Emory University School of Medicine. This inhibitor represents a new paradigm in cancer therapeutics aimed at “intercept” points in mammalian signaling where multiple cell surface receptors converge on single nodal control point e.g. PI-3 kinase or
PTEN. Data will be presented that the inhibition of PI-3 kinase pathways by SF1126 in concert with PTEN inhibition will augment antitumor efficacy.
10:00 Coffee Break, Poster and Exhibit Viewing
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10:45
Implications of Kinase Similarity in Drug Discovery
Dr. Jeffrey Sutherland, Discovery Informatics, Eli Lilly and Company
We have examined the clinical information on kinase inhibitors and analyzed it from small molecule, target and disease perspective. Kinase inhibitors in the clinic are on average of higher molecular weight and more lipophilic than all clinically investigated drugs. Tyrosine kinases from the VEGF and EGFR families are the most pursued targets, and furthermore, oncological indications account for 75% of all
kinase-related clinical interest. In addition, our analysis of the similarity between kinase targets with respect to sequence, selectivity and structure has revealed that kinases having 60% or higher sequence identity are most likely to be inhibited by the same classes of compounds and have similar ATP binding sites. The identification of this threshold, coupled with the widely accepted representation of the sequence-based kinase space is expanding our understanding of the clinical and structural space of the
kinome. |
11:30 Effective Lead Discovery and Lead Optimization: Scherings Kinase Platform
Dr. Ulf Bömer, Head Technology Development & HTS Infrastructure, Research Center Europe,
AD/HTS, Schering AG, Germany
Based on a single assay technology we have built up a whole platform for the fast and cost-effective development and performance of all kinase assays - primary
HTS, compound characterisation and lead optimization - in the 5-µl-scale. It comprises tools for the rapid identification of high-quality peptide substrates and suitable corresponding specific antibodies, and standard procedures for effective automated assay development and broad compound characterisation (mode-of-action studies, selectivity profiling).
12:00 Oral Poster Presentations
12:15pm Lunch on your own
(Technology Workshop Sponsorship Available)
1:45 Binding Assays for Inactive Kinases
Dr. Elma Loomans, Senior Research Scientist, Molecular Pharmacology Unit,
N.V. Organon, The Netherlands
The majority of protein kinase assays used in drug discovery research is enzyme activity assays. These assays measure the phosphorylation of a kinase substrate protein or peptide, which is the end product of the enzyme reaction. Binding assays allow separate measurement of the inhibitory activity of kinase inhibitors on the active and the inactive isoform of the enzyme and do not require ATP or substrate in the reaction. The development and use of high-throughput binding assays for tyrosine and
serine/threonine kinase based on beta-galactosidase enzyme fragment complementation will be presented.
2:15 Screening Kinases using Physiological Substrates
Dr. Karen Kleman-Leyer, Project Leader, R&D, BellBrook Labs
Most kinase assays rely on detection of tagged phosphopeptides, and cannot be easily adapted to use physiological acceptor substrates such as native proteins or lipids. Using a novel kinase HTS assay platform that relies on fluorescent detection of ADP, we demonstrate flexible kinase screening capability using native proteins and lipids as acceptors. In addition, we use the assay to assess the effects of different acceptor substrates on observed inhibitor potency. These studies are designed to lead to more predictive selectivity profiling.
2:45 Technology Hot-Spot
Novel Technologies in Kinase/Phosphatase Assays: Versatile, Homogenous, High Throughput Assay Platforms for Monitoring the Enzyme Activities of Protein Kinase and Phosphatase
Dr. Said A. Goueli, Technology Development Team Leader, Promega Corporation
Because of the significance of protein phosphorylation in normal and abnormal cellular growth and the role the enzymes mediating these processes in the etiology of multiple diseases, we have developed assays that are capable of monitoring the activities of protein kinases and phosphatases in a homogenous and high sensitivity formats to screen for inhibitors of these enzymes in HTS. The presentation will demonstrate the versatility, robustness (Z’>0.7), and rapid performance, and ease of use for various groups of enzymes. The various features of each platform will be highlighted with examples to differentiate the strengths and versatility of each system. The fluorescence based assay is designed to monitor the activities of both kinases ad phosphatases while the luminescent based assay underscores the versatility of using any kinase substrate (protein, lipids, sugars, etc), and thus it is applicable to all kinds of kinase substrates regardless of their nature and with no prior modification. It also detects additional phosphorylation sites of already existing phosphopeptide substrates by enzymes such as GSK-3 and CK1, and monitors the activity of kinases phosphorylating their substrates on multiple sites. Finally both assay platforms are robust as indicated by the high Z’ values (over 0.7), homogenous, can be completed in one step after completion of kinase reaction, does not require antibodies or custom synthesized substrates.
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3:00 Technology Hot-Spot (Sponsorship Available)
PATHTRAKTM Cellular Kinase Solutions: Tracking Compound Effects on Intracellular Signaling Pathways
Dr. Lisa Martel, Scientist, Assay Development, Pharmacology, MDS Pharma Services
One of the most significant hurdles faced by drug discovery programs for kinase targets is obtaining the potency and selectivity needed to move a drug into the clinic. There is a clear benefit to assessing the effects of potential lead compounds on the phosphorylation of endogenous substrates in living cells early on in the drug discovery process. Measuring compound effects on kinase-mediated phosphorylation in whole cells provides the biologically relevant context of intact intracellular signaling pathways and physiological ATP concentrations. To meet the growing needs of the pharmaceutical and biotech industries to profile lead candidates, MDS Pharma Services offers
PATHTRAKTM, a platform of cell-based kinase assays to efficiently assess both potency and selectivity of compounds on intracellular signaling pathways.
3:15 Refreshment Break, Last Chance for Poster and Exhibit Viewing
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Identification + Screening |
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3:55 Chairperson's Remarks
4:00 Kinase Inhibitors: Chemical Genomics Based Screening, Selectivity Profiling, and Mode-of-Action Studies
Dr. Gerard Drewes, Director, Discovery Research, Cellzome Inc., Germany
To predict efficacy and side effects of kinase inhibitors, it is advantageous to determine the target profile directly in the target cell or tissue, as opposed to the more conventional assays with recombinant enzymes. We have developed a range of Chemical Genomics methods that allow us to study the interaction of compounds with their targets directly in tissue culture or even tissue samples from patients. The drug or lead compound is applied at a range of concentrations to living cells or to cell
lysates, and the lysate is subsequently captured on a proprietary affinity matrix which specifically binds hundreds of protein kinases and
kinase-associated proteins. The target affinity profiles of the compound are then determined by differential analysis of the captured proteins from treated and untreated samples, by quantitative mass spectrometry using stable isotopes. The application of this technology to library screening, selectivity profiling, and mode-of-action studies on compounds will be presented.
4:30 Concept of Focused Diversity: Application to Development of Specific and Dual Inhibitors of VEGFR-2 with in vivo Activity
Dr. Alex Kiselyov, Executive VP of R&D, ChemDiv, Inc.
Goal: to develop a robust algorithm for identification of novel specific and dual inhibitors of VEGFR2, the key protein modulating
angiogenesis. In order to achieve our goal, we selected a subset of compounds (Focused Diversity: 3,000
cmpds/275 templates) pre-screened against a number of diverse biological targets. Specifically, this set contained molecules with confirmed in vitro/cell-based activities against:
i) “traditional” (kinases, GPCR's, ion channels) and ii) “unconventional” targets
(Hh/Wnt signaling pathways). Initial assays yielded 11 novel
chemotypes, all showing good in vitro and cell-based activities (< 1
uM) against VEGFR1/2. MedChem follow-up furnished actives with cell-based potencies
(phosphorylation, functional assays) of 25-100 nM. Compounds displayed favorable in vitro
PK: good absorption (Caco2), hhep stability, water/buffer solubility (> 5
mM) and plasma binding (< 75%). Lead candidates displayed promising oral exposure in murine models (ca. 15-20
uM/4 hrs; acute dose of 10 mg/kg, t1/2 = 6-9 hrs), MTD (acute/chronic PO's> 250 mg/kg) and efficacy in tumor xenografts (HT29) to warrant further extended in vivo studies. Conclusion: focused diversity concept has been validated in the identification of novel efficacious inhibitors of VEGFR2s.
5:00 Panel Discussion
5:45 End of Day
For more information, please contact:
Margit Eder, Ph.D., Conference Director
Phone: 781-972-5478 • E-mail: meder@healthtech.com
For exhibit and sponsorship information, please contact:
Suzanne Carroll, Manager, Business Development
781-972-5452 • scarroll@healthtech.com
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