TUESDAY, JUNE 24
7:30am Morning Coffee
8:30 Chairperson’s Keynote Introduction
KEYNOTE PRESENTATION
8:40 Mechanism – Driven Approaches to Molecularly Targeted Cancer Drug Discovery
Robert A. Copeland, Ph.D., Vice President, Biology, Oncology Center of Excellence in Drug Discovery, GlaxoSmithKline Pharmaceuticals
Oncology drug discovery at GSK is focused on molecularly targeted therapeutics for the treatment of cancer and for cancer supportive care indications. A guiding principle of our approach is that a detailed, mechanistic understanding of pathobiology and of drug action – at the molecular, cellular and organismal level – is essential for effective drug discovery. Towards this end we have buildt a discovery organization that encompasses robust biochemical, cellular biological and in vivo pharmacological interrogation of drug candidates. Examples of the application of these approaches to several cancer drug – seeking efforts will be presented.
Screening and Lead Optimization
9:25 Discovery and Development of Selective, Orally Bioavailable Tyrosine Kinase Inhibitors for Targeted Treatment of Human Cancers
Stephen K. Burley, M.D., D.phil., F.R.S.C., Chief Scientific Officer and Senior Vice President, Research, SGX Pharmaceuticals, Inc.
The SGX platform utilizes high-throughput X-ray crystallography to guide fragment-based lead identification and subsequent optimization of potency, selectivity, and other drug-like properties. Biologically active compound series are prioritized for further medicinal chemistry and preclinical development efforts using the results of in vitro and in vivo ADME and in vitro toxicology studies. Recent progress with inhibitors of the MET receptor tyrosine kinase (SGX523—Clinical; SGX126—Preclinical) and of drug-resistant BCR-ABL (SGX393—Preclinical) will be discussed. MET represents a potentially important target across a wide range of cancer indications, wherein a personalized-medicines approach may prove particularly beneficial. BCR-ABL is the target for treatment of chronic myeloid leukemia, for which approved 1st and 2nd generation inhibitors are encountering resistance in patients.
10:05 Networking Coffee Break, Poster and Exhibit Viewing
10:45 Screening for plk1 Inhibitors
Michael Yaffe, Ph.D., Associate Professor, Biology, Massachusetts Institute of Technology
11:15 Rapamycin-Derived mTOR Inhibitors by Synthetic Biology
Ming-Q Zhang, Ph.D., FRSC, Senior Vice President, R&D, Biotica Technology Ltd.
mTOR is one of the most important kinases in our search for anticancer and anti-inflammatory medicines. The impact of PTEN deficiency on the PI3-Akt-mTOR pathway, the interplay between the 2 complexes of mTORC1 and mTORC2, and the newly discovered role of FKBP38 in antagonizing mTOR have all had major influence on our understanding and effective approaches to modulate this kinase. A series of novel rapamycin derivatives have been generated by a synthetic biology approach. SAR analysis of these rapamycin derivatives on some of the cellular readouts related to the mTOR pathway will shed some light on differentiation of these closely related analogues from their natural product lead.
| 11:45 Development of an HTRF Primary Assay for a Kinase Using Selective Labeling Methods |
Sponsored by
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Weidong Ding, Ph.D., Principal Scientist, Biochemical and Cellular Sciences, Screening Sciences
Wyeth Research
An HTRF homogeneous assay is desirable for HTS of a kinase in that it eliminates wash steps that are time-consuming. The use of D2 as a fluorophore is especially helpful because the labeled proteins can be readily characterized by LC-MS due to the small size of D2. The D2 loading and distribution on the proteins were correlated with HTRF signals, resulting in improved labeling conditions. We describe here the methods of selecting derivatives of protein substrate that led to higher degree of D2 labeling and better HTRF signals.
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| 12:15pm Lunch and Learn Tools for the Interrogation of Lipid Kinases and the PI3K/AKT/mTOR Signaling Pathway |
Sponsored by
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Sara B. Hereley, Product Manager, Invitrogen
The PI3K/AKT signaling pathway is fundamental for many aspects of cell cycle, growth, death and survival, and plays a pivotal role in the genesis of numerous diseases, including cancer and diabetes. Because of the complexity of this signaling pathway, especially as applied to the regulation of the mammalian target of rapamycin (mTOR), a variety of methods will be critical for the proper identification of small-molecule mediators of this pathway. To address these needs, we have developed a suite of fluorescence-based tools for the study the components of this pathway, in formats ranging from purified biochemical assays to cellular assays to monitor global pathway activity. |
1:40 Chairperson’s Remarks
1:45 Three-Dimensional (3D) Overlay Culture Models of Human Breast Cancer Reveal a Critical Sensitivity to MEK Inhibitors
Raymond R. Mattingly, Ph.D., Associate Professor of Pharmacology, Wayne State University
We have screened drugs that inhibit several pathways downstream of Ras activation, including MAP kinase, PI 3-Kinase, Rac1 and Rho/myosin light chain kinase (MLCK) for their effects in both traditional, two-dimensional (2D) cell culture and overlay 3D cultures in reconstituted basement membrane (rBM). We tested the agents against a model of the isogenic progression of human breast cancer, using spheroid size to measure cell growth in 3D overlay culture, and MTT assay to determine proliferation in 2D culture.
Remarkably, we find a selective effect of the MAP kinase kinase (MEK) inhibitors to block the proliferation of the more transformed cells in the 3D overlay cultures, whereas in 2D cultures the inhibitors produced a similar reduction of proliferation in all the cell lines tested. In this study, we found little reduction in growth in the 2D or 3D overlay cultures in response to PI3 kinase, Rac, or MLCK inhibition. The results from this study suggest that 3D overlay culture in rBM may be a good model for the screening of Ras pathway inhibitors, with sensitivity to cancer progression status that is superior to that in traditional 2D culture models.
2:15 Substrate Profiling of Protein Kinase C Isoforms Using Serine/Threonine Dynamic Peptide Microarrays for Design and Synthesis of Isozyme Selective Bisubstrate Based Inhibitors
Rob M. J. Liskamp, Ph.D., Professor, Medicinal Chemistry and Chemical Biology, Utrecht Institute for Pharmaceutical Sciences, Utrecht University
The development of a novel non-radioactive method for detection of phosphorylation using porous micro arrays is described. The used dynamic arrays are decorated with known kinase peptide substrates containing serine/threonine phosphorylation sites. These peptide micro arrays allowed accurate and reproducible profiling of different PKC isozyme activities towards multiple substrates in one experiment. The found peptide substrate phosphorylation sites sequences served as a starting point to develop bisubstrate inhibitors leading to isozyme-specific inhibitors.
2:45 Tailoring Computational Methods for Kinase Targets
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Sponsored by
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Dipesh Risal, Ph.D. Product Manager, Life Sciences, Accelrys Inc.
Accelrys offers a suite of programs and tools that are tailored for structure-based design projects involving protein kinases. Several workflows and case studies will be presented that involve not only the docking and scoring against the flexible binding sites of representative kinases, but also tools that allow a visual exploration of the kinome in an
intuitive context.
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| 3:00 Kinase-Glo® Assays Platform: A Robust Bioluminescent System for Monitoring the Activities of all Types of Kinases |
Sponsored by
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Said A. Goueli, Ph.D., Research Fellow, Cell Analysis Group, Research and Development, Promega Corp.
The luminescence based Kinase-Glo assay platform has gained wide acceptance in many drug screening programs. The assay measures ATP depletion, a general feature of all kinases, as a read-out for kinase activity. Thus it is applicable to most kinase-substrate combinations with no prior substrate modification. Examples of its use with several protein kinases using peptides as well as protein substrates will be presented, as well as examples of its use with some non-protein kinases (lipid kinases, sphingosine kinases, ceramide kinases, and glucokinases). The assay can also be used with kinases that use pre-phosphorylated substrates such as GSK-3ß and CK1, or with kinases that phosphorylate on multiple sites on a substrate. Data showing the use of the assay with higher concentrations of ATP (up to 500µM) will also be presented. Utilizing higher ATP levels maximizes the probability of selecting non-ATP binding site inhibitors during screening. Data comparing Kinase-Glo to other luminescent and TR-FRET based assays will also be presented. |
3:15 Networking Refreshment Break, Poster and Exhibit Viewing
4:00 Discover Slowness: Enhancing Drug Efficiency Through Inhibitors with Long Kinase-Drug Residence Times
Doris Hafenbradl, Ph.D., Executive Vice President, Screening & Proteins, Proteros Biostructures
Retrospective analysis has shown that amongst the successful kinase drugs a surprisingly large proportion show slow drug-kinase binding kinetics. Broader analysis reveals that this phenomenon is not only true for kinase drugs but also for drugs throughout all target families. Despite this clear advantage of slow binding drugs no systematic development of drugs with this property is happening so far. This discrepancy is mainly due to the lack of time and cost efficient methods to measure kinase binding kinetics for large numbers of drug candidates. Here we present a novel technology that opens this bottleneck and allows the rational design of drug candidates with slow kinetics. Moreover we discuss how this new technology can be implemented into the current lead optimization process in order to produce better drugs.
4:30 Panel Discussion: Screening for Gold – Do we Need More Targets?
5:30 End of Day Two