Tuesday, June 8

5:00 pm-6:00 pm Early Registration, Poster and Exhibit Setup

Wednesday, June 9

8:00 Registration, Morning Coffee, Poster and Exhibit Setup

Functional Understanding of Kinases

9:00 Welcome by Session Chairperson
Dr. Mitchell Kostich, Discovery Technology, Schering-Plough Research Institute

9:15 Census and Classification of Human Protein Kinases
Dr. Mitchell Kostich
Recent work suggests that there are more than 500 protein kinase genes present in the human genome. Creating an accurate census and construction of a phylogenetic tree for this family are important early steps towards determining which family members are attractive drug targets and which should be considered in counter screening. Efforts by several groups to enumerate and classify members of this gene family are summarized and reconciled with one another. Special attention is given to unresolved issues and application of this work to drug discovery.

9:45 Tracking the Kineome by Multiblotting with Antibodies and Peptide Antibody Mimetics (PAM's)
Dr. Steven Pelech, President and CSO, Kinexus Bioinformatics Corporation
With a vast suite of highly validated antibodies, the Kinetworksä multiblotting service has been successfully used to track more than 130 protein kinases and 135 known phosphorylation sites in hundreds of human tissues and cells and other experimental model systems. Kinexus has also begun to develop high affinity, specific peptide antibody mimetics (PAM's) to eventually monitor all of the known human protein kinases with peptide macroarrays. In the near future, this data will become Internet accessible and used to deduce the composition and architecture of protein kinase networks.

10:15 Kinases In Bulk: Gene Families As System Biology Tools For Functional Studies
Dr. Liang Cao, Director of Cell Biology, OriGene Technologies, Inc.
Protein kinase genes are known to play key roles in cellular processes including cell division, signal transduction, apoptosis and cell mobility. Because dysfunction of kinase genes results in a variety of diseases, such as cancer, diabetes and heart disease, this family of genes has captured academic and pharmaceutical research interest. The Kinase CloneSetä, composed of more than 460 full-length kinase genes, approaches completion of the predicted 518 putative genes in the "Kinome" (Manning, G. et al., Science 298:1912, 2002). The Kinase CloneSet provides researchers the necessary tools for high throughput screening and functional family studies. In this presentation, experimental results from over-expression and expression knock-down of kinases in cancer cells, using the Kinase CloneSet, will be presented.

10:45 Poster and Exhibit Viewing, Coffee Break

11:15 Global Analysis of Yeast Protein Kinases using Protein Chips
Ms. Geeta Devgan, Department of Molecular, Cellular, and Developmental Biology, Yale University
We are currently using proteomic methods to understand cellular signaling pathways and protein-protein interactions of the budding yeast, Saccharomyces cerevisiae. 121 of 122 protein kinases were cloned and purified from yeast as GST fusions and analyzed for their ability to phosphorylate 20 different yeast substrates. We have also constructed a protein chip, an array of 5800 proteins overexpressed from yeast, which can provide a global analysis of kinase activity over the entire yeast proteome. Once substrate specificity is determined, we can begin to explain redundancy between various kinases and further explore their mechanisms of action. Co-Authors: Heng Zhu, Jason Ptacek and Michael Snyder

11:45 Protein Micro Arrays as a Tool to Identify Novel Protein-Protein Interactions
Dr. Ulrike Korf, Molecular Genome Analysis, German Cancer Research Center (DKFZ),
A variety of assays have been established for functional characterization of novel human ORFs, e.g. microarray-based kinase assays. We want to apply this assay platform to explore in vitro signaling properties these novel human proteins are potentially involved in. Minute amounts of recombinant purified proteins are dispensed in a micro array format on a 3-dimensional surface using a non-contact spotter (BioChip Arrayer, Perkin Elmer). Currently we have adapted 14 different kinase assays to a miniaturized assay format. All assays are easily adaptable to a high-throughput format. Assays are carried out on the basis of probing protein-protein interaction using fluorescently labeled kinases as well as on kinase-catalyzed incorporation of a radioactive [P-33] phosphate into arrayed proteins. Candidate proteins revealing a distinct interaction and/or phosphorylation pattern are subjected to further characterization to confirm the detected novel protein phosphorylation by mass spectrometry. Co-Authors: Simone Schleeger, Christian Schmidt, Dirk Bossemeyer, Peter Drückes, Thorsten Kohl, Barbara Ueberle, Thomas Faupel, Konrad Büssow, Martina Schnoelzer, Stefan Wiemann, and Annemarie Poustka.

12:15 Activating Mutations in the EGF Receptor underlie the Clinical Response to Gefitinib
Dr. Jeffrey Settleman, Associate Professor, Medicine, Harvard Medical School -MGH Cancer Center
Approximately 10% of patients with non-small cell lung cancer exhibit a rapid and substantial clinical response to the specific EGF receptor inhibitor, gefitinib (Iressa). We found that tumors from 8 of 9 of such “responders” (and none of 7 “non-responders”) harbor somatic mutations within the catalytic kinase domain of the EGF receptor that render them “activating” as well as highly sensitive to the inhibitory actions of gefitinib. Thus, screening for such mutations in lung cancer patients should be very useful in identifying patients that are likely to respond to gefitinib therapy. Co-Authors: Dr. Thomas Lynch, Dr. Daphne Bell, Dr. Dr. Daniel A Haber, and Dr. Raffaella Sordella

12:45 Luncheon Workshop Microfluidics: High Quality Screening From Targets to Leads Presented by Dr. Seth P. Cohen, Director: Application Sciences, Caliper Life Sciences

Sponsored by:

Microfluidics-based assays have been deployed throughout the drug discovery process from assay development for novel enzymes through to lead optimization of pre-clinical drug candidates. One of the typical hallmarks of microfluidics applications is the overall higher quality of the data compared with microplate-based formats. The basic principles of LabChip technology will be described using examples of commercially available microfluidic assays for phosphatase, protease and kinase targets. A variety of tools and applications have been developed to speed the discovery of protease and kinase substrates as well as to reduce assay development timelines. Examples in which the analytical quality data produced using LabChip technologies identifies hits traditional technologies might overlook will be described and the application of a panel of kinase assays for use in lead profiling will be presented. In addition, data will be presented on the development of novel primary screening assays for lipid modifying enzymes as well as cell-based GPCR assays.

Drug Development: Screening Strategies

1:55 Comments by Session Chairperson
Dr. Geoffrey Alms, Upstate, Inc.

2:00 Screens for Novel Small Molecule and cDNA Inhibitors of PI3K/Akt Signaling
Dr. Charles Cho, Postdoctoral Associate, Chemistry, Scripps Research Institute
The FOXO family of transcription factors are mediators of various cellular functions including cell cycle progression and cell death, and are negatively regulated by activation of the PI3 kinase pathway. Akt phosphorylation of FOXO family members leads to FOXO sequestration in the cytoplasm and transcriptional inactivation. Using high throughput imaging of FOXO nuclear translocation, we have screened both small molecule and arrayed cDNA libraries and discovered novel inhibitors of PI3 kinase signaling in both classes.

2:30 High Content Screening Based on Imaging of Translocation of a Key Transcription Factor
Dr. William R. Sellers, Assistant Professor of Medicine, Harvard Medical School, Dana-Farber Cancer Institute
Summary Unavailable at Printing

3:00 Evaluating Kinase Compound Selectivity: Testing a Panel of Public Compounds
Dr. Geoffrey Alms, Senior Applications Scientist, Phosphorylation Team, Upstate Inc.
In order to further understand the selectivity of kinase inhibitors both publicly released and those in the clinic, a panel of 60 compounds has been screened against 101 kinases representing a diverse cross section of the kinome. We will present our thoughts and conclusions from this study as well as ideas about meeting the future needs of kinase selectivity screening.

3:30 Poster and Exhibit Viewing, Refreshment Break

4:00 Scaffold Based Drug Discovery against Kinase Targets
Dr. Michael V. Milburn, Senior VP of Research, Plexxikon Inc.
We will demonstrate how, through a combination of biochemical assay screening of a low molecular-weight 20K compound collection and high throughput co-crystallographic structure analysis of bioactive hits co-complexed with kinases, we identify multiple low molecular weight starting points for lead discovery. Using the available co-structure information, discovery chemistry synthesizes additional compounds at multiple points of R-group substitution for lead optimization and co-crystallization with the target. Those scaffolds that are novel chemistries, chemically tractable, and best suited for drug discovery are further developed to yield proprietary small molecule drug leads for pharmaceutically relevant protein targets. We will show several examples of this technology applied to multiple kinase drug discovery partnerships with other companies as well as Plexxkon's internal kinase drug discovery efforts. In general, kinase drug leads developed with this scaffold-based approach will be shown to have good ADME properties as well as in vivo efficacy.

4:30 Kinases, Homology Models, and High Throughput Docking
Dr. David J. Diller, Head, Molecular Modeling, Pharmacopeia, Inc.
Over the past few years we have undertaken the task of validating the use of homology modeling in the context of kinase targeted drug design. In this talk we briefly discuss the validation studies. Furthermore, we discuss how the results have then been used in areas such as library design and lead optimization.

5:00 Close of Day One