Tuesday, June 8

Structural and Bioinformatics
Approaches

9:00 Comments by Session Chairperson
Dr. Kenneth Lundstrom, CSO, BioXtal

9:15 Towards an Artificial Nose Sensor Based on Olfactory GPCR's
Dr. Claire Komives, Department of Chemical and Materials Engineering, San Jose State University
A mathematical model of olfactory GPCR signal transduction has demonstrated that varying the ratio of GTP/GDP in the sample buffer can enable the development of olfactory receptor-based chemical sensors with up to six orders of magnitude of dynamic range. Experiments with receptors from rat noses have validated the model and we have begun the cloning of GPCRs from a mouse cDNA library (donated from Richard Axel's lab). Additionally, experiments on the expression of the OR5 receptor (gift of Prof. Hans Breer) in CHO cells are currently underway, exploring the use of a novel membrane trafficking protein.

9:45 Small Molecule Antagonist of Thrombin-Mediated PAR1/Gq Signaling
Dr. Annette Gilchrist
Many biological processes result from activation of G protein coupled receptors (GPCRs) and their signaling partners, the G proteins. However, many GPCRs, including the protease activated receptor (PAR-1) have been refractory to traditional methods of identifying small molecule antagonists. PAR-1 is promiscuous, coupling to multiple G proteins in the same cell. To elucidate which G protein underlies a specific thrombin-mediated response, we introduced Galpah C-terminal minigene vectors into cells. Results indicate that the different Galpah subunits have unique roles. We went on to identify small molecule antagonists of thrombin-mediated PAR-1/Gq signaling. The small molecules were tested for their ability to affect Gq binding to PAR-1, as well as thrombin-mediated calcium flux and platelet aggregation.

10:15 Mimetics: From Ligand to Drug
Dr. Peter Cassidy, CEO, Mimetica
GPCR binding peptide ligands often recognize reverse-turn containing conformations, and this information is potentially useful in the drug development process. Mimetics of known ligands can be made, directly giving drug-like leads, and screening libraries for orphan receptors can be designed around the same mimetic scaffolds. As success depends on the accuracy if the mimetics, their ease of synthesis and whether they are sufficiently drug-like, our work in the area will be described.

10:45 Poster and Exhibit Viewing Coffee Break

11:15 Structural Genomics Approaches to Membrane Proteins
Dr. Kenneth Lundstrom
Less than 1% of the more than 20,000 structures deposited in public databases are from membrane proteins. This is due to the complexity of membrane proteins and therefore their overexpression, purification and crystallization is by far more difficult than for soluble proteins. On the other hand, membrane proteins represent some 70% of the current drug targets. Large networks have been established in USA, Japan and Europe to develop novel technologies in all areas from overexpression to crystallography in attempts to obtain novel high resolution structures for targeted drug design. Many of the networks deal only with prokaryotic membrane proteins, because of higher success rate for these targets. Several networks have included a few mammalian targets in parallel to bacterial targets. The MePNet program, however, has uniquely concentrated on eukaryotic GPCRs, mainly of human origin. The progress of expression, purification and crystallization activities in the various networks will be presented.

11:45 Design Strategies for GPCR-Focused Libraries
Dr. Patrick Jimonet, Chemical Biology GPCR, High Throughput Medicinal Chemistry, Aventis Pharmaceuticals
Strategies to design GPCR-focused libraries are progressively moving from a purely compound-based approach to a more sophisticated recognition-based approach. Our own initiative has been the design of a dynamic knowledge-based GPCR compound collection, regularly improved by libraries focusing on receptor subfamilies or bioinformatics clusters. Informatics tools have been acquired and/or customized to leverage knowledge linking biological and chemical spaces. Our integrated strategy and other recent approaches will be presented.

12:15 Panel Discussion with all Morning Speakers

12:30 Lunch on Your Own

Signaling and Disease Applications

1:55 Comments by Session Chairperson
Dr. Annette Gilchrist, Northwestern University

2:00 Solid-state NMR Studies of Transmembrane Signaling
Dr. Lynmarie K. Thompson, Associate Professor, Department of Chemistry, University of Massachusetts
Solid-state NMR is a promising approach for structural studies of membrane proteins, that can be used for example to measure local structure at the ligand site of a receptor. We are investigating the molecular mechanism of transmembrane signaling in a well-studied model receptor system, the bacterial chemotaxis receptor. Prior studies of bacterial chemotaxis receptors have yielded structures of the external and internal domains and have led to proposals that ligand binding causes subtle conformational changes that are transmitted across the membrane to control the phosphorylation activity of an associated kinase. As a direct test of this model, it is critical to measure the proposed conformational changes in the intact, membrane-bound receptor. We have developed site-directed solid-state NMR as a novel approach for obtaining high-resolution distance measurements for this purpose. Our results have provided the first high-resolution measurements of the structure and ligand-induced structural change in the intact, membrane-bound receptor. These site-directed solid-state NMR approaches should prove valuable for probing local structure and mechanisms of other important membrane receptors.

2:30 Receptors for Lipid Mediators: Role in Inflammation and Metabolic Disease
Dr. Bjorn Olde, Associate Professor, Molecular Neurobiology, Lund University
The recently identified free fatty acid receptor, FFA1R (previously known as GPR40) is activated by medium- to long-chain FFAs. It seems to play a key role in insulin secretion and etiology of type2 diabetes. It is a hitherto unknown target for glitazone-type antidiabetics. FFA2R (previously, GPR43) is activated by short FFAs, is expressed in immune cells, and has an anti-migratory effect. Thus, an effect which is "opposite" to that of the leukotriene B4 receptor, BLT. These lipid-mediator receptors represent novel drug targets involving hitherto unknown action mechanisms.

3:00 Poster and Exhibit Viewing, Refreshment Break

3:30 Development of Drugs Acting at GPCRs for the Treatment of Cachexia and Obesity
Dr. Val S. Goodfellow, Associate Director, Medicinal Chemistry, Neurocrine Biosciences, Inc.
Numerous GPCRs in the central nervous system are involved in the regulation of food intake, metabolism and body weight. We have targeted the MC4 receptor for treatment of cachexia and the MCH receptor for treatment of obesity. Our current understanding of these receptors, their endogenous ligands and the development of compounds from high-through-put screening hits to potent ligands with drug like properties adequate for proof of principle studies in feeding disorders will be discussed.

4:00 Discovery of Novel GPCR Targets Through a Reverse Genetics Screen for "Druggable" Gene Function in Mice
Dr. Barry Kenny, Commercial Director Paradigm Therapeutics Ltd
We have identified novel GPGR targets with therapeutic utility in the treatments of pain and modulation of the GnRH sex hormone axis. Detailed in vivo studies reveal the direct relevance and mechanistic basis for these genes as drug targets, with additional studies confirming the relevance to human disease. These targets represent novel drug discovery opportunities in key areas of unmet medical need. We have constructed a high throughput reverse genetics screen to determine the function of biological orphans of mammalian genes of special interest in the discovery of new drugs. We shall describe our screen, discuss some of the phenotypes obtained, and highlight some of the detailed in vivo profiling that has uncovered key GPCR drug targets using this approach. Key programs in the areas of pain and neuroendocrinology will be described. In the latter case, the relevance and linkage to human therapeutics will demonstrate how we have been able to identify a novel approach to modulation of the GnRH axis with important therapeutic implications.

4:30 Panel Discussion with all Afternoon Speakers

5:00 Close of Conference

Call for Sponsors and Exhibitors 
Showcase your company's expertise, brand your solutions and develop revenue opportunities with qualified decision-makers by becoming an exhibitor or sponsor at GPCRs and Kinases. Sponsorship opportunities include: technology workshop presentations, poster competitions, refreshment breaks and networking receptions just to name a few. We also have high-visibility items such as conference padfolios and badge lanyards, which will brand your company name and logo.

For more information on exhibiting and sponsoring, please contact Arnie Wolfson at 781-972-5431, or awolfson@healthtech.com