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This is the agenda for the 2012 meeting. Please check back for an updated version!
Day 1 Day 2 Day 3
Day 1 - Wednesday, October 24, 2012
Plenary Keynote Session I - 8th Modern Drug Discovery & Development Summit
Moderator: Immanuel Freedman, Manager, Pharmacometrics, GlaxoSmithKline
1:30 Clinical Biomaterials for Regenerative Medicine: From Bench to Business

Glenn D. Prestwich
Presidential Professor
Director, Therapeutic Biomaterials Center
Special Presidential Assistant for Faculty Entrepreneurism
Medicinal Chemistry
University of Utah
Faculty and student entrepreneurs working together maximize scholarly and societal impact. First, I will describe the entrepreneurial ecosystem at the University of Utah. Then, I will describe a case study for commercialization of a university technology in the area of regenerative medicine.

Clinical biomaterials for regenerative medicine. Injectable and biocompatible vehicles for delivery, retention, growth, and differentiation of progenitor cells are needed for cell therapy. We created a synthetic extracellular matrix (sECM) from hyaluronic acid (HA) that affords highly reproducible, manufacturable, approvable, and affordable biomaterials. The in situ crosslinkable sECM hydrogels can be customized for use with progenitor and mature cell populations obtained many tissues, including skin, fat, liver, heart, brain, muscle, bone, and cartilage. In addition, sECMs have been developed for rapid expansion and recovery of cells in 3-D, and for the bioprinting of engineered tissue constructs. The technology is being commercialized in three fields of use: human medical devices, cell therapy and research tools for 3-D cell culture, and veterinary wound care and adhesion prevention.
2:15 Building a New Ecosystem for R&D with Academia and Pharma: Centers for Therapeutic Innovation


Anthony J. Coyle
Vice President and Chief Scientific Officer
Centers for Therapeutic Innovation
Pfizer
In 2010, Pfizer took decisive steps to launch the Centers for Therapeutic Innovation and, in doing so, has led the industry in innovation around early R&D models. Since then, CTI has made considerable operational progress - building a strong team, establishing four US-based sites, signing 19 academic partners, and generating an early research portfolio. CTI is poised to be a transformational force, employing an entrepreneurial R&D model that accesses the best science in the world – regardless of geographic origin – to deliver mechanistically-relevant clinical studies that can translate into differentiated, clinically-validated candidates that align with the company’s strategy in order to deliver important medicines to patients. This year’s plan focuses on our approach to building an innovative early R&D portfolio that can help create options for the other RUs and BUs across Pfizer. Importantly, precision medicine is embedded into the CTI approach, in order to deliver data-rich packages to the RUs/BUs for their consideration. Working jointly with academic medical centers, and their unparalleled access to well-phenotyped human tissue samples, CTI projects can aggressively pursue patient stratification analyses and clinical strategies early in development. CTI is focused on identifying the best science in the world, demonstrating scientific and operational excellence in progressing the portfolio, and building strategic alignment around projects with our RU and BU colleagues.
3:00 Networking and Refreshment Break
3:30 Innovation Through Cutting Edge Translational Medicine & Virtual Pharma Model to Transform R&D Productivity



Harsukh Parmar
Vice President, Global Head
Translational & Experimental Medicine, Inflammation
Roche
R & D productivity remains poor despite continual increase in R & D budgets. Innovation has often become stifled in large companies by huge demands on unrealistic target product profiles (TPP’s), self-perpetuating huge bureaucracies, inadequate human target validation, excessive spend on easily tractable targets, incapable of delivering differentiated products, and poor leadership in decision making at all phases of R & D, importantly at the target identification/target validation discovery stage, but also at other milestones. These deficiencies lead to large excessive spend of R & D budgets on projects which get into human clinical trials but which have no future and which have no chance of delivering the agreed TPP’s wanted by the strategic marketing organizations. Currently marketed products, which are now becoming generic, continue to pose high hurdles on efficacy and safety and benchmarking against these products is often poor in early stages of discovery and early development. Failure on efficacy grounds in Phase IIb and Phase III is still very common and is unacceptable from both an ethical and R & D budget perspective. Sales & Marketing spend still remains higher than R & D spend across the industry and this needs to be reversed as soon as possible. This model needs to change. True globalization, better “human target validation”, stronger input from translational medicine, excellence in visionary leadership who truly understand R & D, and transformation into a virtual pharma model are all viable options to address some of these productivity challenges.
4:15 Leveraging Partnerships to deliver Precision Medicine


Morten Sogaard
Executive Director and Head
Biotechnology and Precision Medicine External R&D Innovation
Pfizer
Health care and Pharmaceutical R&D today is expensive, reactive and often ineffective – largely because we don’t understand the diseases and patients we are treating as well as we would like to. We need to more precisely measure patient disease phenotypes and correlate it with molecular markers to better understand disease and effect of drug treatment. We term the application of this approach of integrating clinical and molecular information to drug discovery Precision Medicine R&D

The key value proposition for PM is to increase the therapeutic index by treating only those patients likely to respond or excluding those most likely to experience side effect. In a best case scenario smaller, cheaper and faster clinical trials, and earlier submission and launch. More dramatic therapeutic effects drive greater value for patients and easier to convince the value of our medicines to payers.

The talk will give examples of how Precision Medicine and diagnostics are applied in Pfizer R&D projects. Particular emphasis will be put on innovative partnerships and why a open R&D Ecosystem will be essential to the success of Precision Medicine.

More information can be found in Dolsten & Søgaard: Precision medicine: an approach to R&D for delivering superior medicines to patients. Clinical and Translational Medicine 2012, 1:7

Benefits:
• Overview of Precision Medicine R&D concept
• Examples of Application of Precision Medicine R&D in Pfizer Projects
• Examples of Pfizer Precision Medicine R&D partnerships
• Overview of how genetics and omics will contribute to success of PreM R&D
Day 1 Day 2 Day 3
Day 2 - Thursday, October 25, 2012
7:00 Continental Breakfast & Registration
7:55 Welcome & Opening Remarks
Plenary Keynote Session II - 8th Modern Drug Discovery & Development Summit
Moderator: Ben Zeskind, Co-Founder and CEO, Immuneering Corporation
8:00 DNA Encoded Libraries – A Disruptive Innovation for Molecular Discovery?


Barry Morgan
Vice President
Molecular Discovery Research
GlaxoSmithKline
Encoded Library Technology (ELT) is a novel approach to molecular discovery based upon the creation of large encoded libraries of novel, drug-like structures that can be rapidly interrogated to identify families of compounds with affinity for a macromolecular target. The utility of libraries assembled by combinatorial chemistry to molecular discovery has been constrained due to deconvolution limitations: ELT addresses this issue by encoding each molecule with a covalently attached DNA sequence. The resulting libraries are screened by “selection” on the basis of affinity, “hits” identified by high capacity DNA sequencing, and the corresponding organic structures synthesized and assayed.

ELT Libraries are constructed by sequential “split and pool” cycles that alternate organic synthesis with enzymatic oligonucleotide ligation: in this way we have been able to assemble a library pool containing over 30 billion (3x10^10) structures. I will describe studies with these libraries against a variety of drug targets, yielding families of “hit” molecules that inhibit the function of these targets with single digit nanomolar potency.

Biochemical combinatorial techniques such as phage display, RNA display and aptamers have proven to be reliable methods for generation of ligands to protein targets. Adapting these techniques to small synthetic molecules has been a long-sought goal. We believe that ELT represents the attainment of that goal.
8:45 Catalyzing Innovation: The NIH National Center for Advancing Translational Sciences


John C. McKew
Chief Therapeutics Development Branch, Therapeutics for Rare and Neglected Diseases (TRND) BrIDGs (former NIH-RAID); Division of Preclinical Innovation
National Center for Advancing Translational Sciences [NCATS], National Institutes of Health
The National Center for Advancing Translational Science (NCATS) is a newly formed center within the National Institutes of Health. It was created by merging existing programs and several new initiatives into a new center. This new center now has two main divisions: the Division of Preclinical Innovation and the Division of Clinical Innovation. The Division of Preclinical Innovation encompasses the former NIH Center for Translational Therapeutics and provides a unique range of programs addressing many aspects of therapeutics development. The programs include the NIH Center for Chemical Genomics (NCGC) which is one of the Molecular Libraries screening centers; the Tox 21 programs; a multiagency collective in vitro toxicology screening program, Therapeutics for Rare and Neglected diseases program; a collaborative drug discovery and development program focused on preclinical to early clinical development of candidate therapeutics for rare and neglected tropical diseases, and Bridging Interventional Development Gaps (the former NIH-RAID program); a collaborative late preclinical development resource program. The division of clinical innovation is currently comprised of the clinical translational science awards. This talk will give an overview of the new center and highlight the translational research programs contained within.

Benefits:
• Better understanding of the role the NIH and NCATS play in developing novel public private partnerships for therapeutic development
• Understanding of the existing and newly developed preclinical development initiatives NCATS has announced along with their solicitation timelines.
• Examples of successful collaborative projects from the Therapeutics for Rare and Neglected Diseases (TRND) and Bridging Interventional Development Gaps (BrIDGs) program portfolios.
9:30 Morning Networking Break
Label Free Technologies
Moderator: Frederik Sunberg, Global Director, Life Sciences, GE Healthcare
10:00 Using Label-free Technology Solutions to Improve Drug Discovery and Development
Fredrik Sundberg, Global Director, Life Sciences, GE Healthcare
The implementation of enabling technology solutions can significantly impact on critical stages of drug discovery and development, improving both drug candidate quality and overall productivity by reducing safety risks and improving efficacy.

The use of innovative technologies, such as label-free assays, can rapidly eliminate false-positive hits in screening and drive lead optimization to improve the probability of success in later clinical stages. For example, the implementation of rapid SPR biosensor assays provides more information on protein interactions that improve decision-making in both hit validation, lead selection, pre-clinical and clinical safety assessment.

The objective of the presentation is to describe label-free assays and how they can support prediction of safety and efficacy in key applications throughout the workflow. Key applications for both small molecules and bio-therapeutics from early discovery to clinical development will be addressed, such as:
• Screening and hit validation
• Lead selection and optimization
• Immunogenicity monitoring
• Protein structure-function mapping

Key benefits:
• Learn how to use label-free assays to:
• improve productivity
• get faster time-to-result
• monitor biomarkers for safety
• characterize protein drugs, MABs and biosimilars
10:25 FEATURED PRESENTATION
Label-Free Approaches Enabling Hit Identification and Lead Validation


George Addona
Executive Director
In Vitro Pharmacology
Merck Research Laboratories
11:00 Label-free, Immobilization-free Interaction Studies Using Microscale Thermophoresis
Stefan Duhr, CEO, NanoTemper Technologies GmbH
The analysis of bio-molecular interactions, such as protein-protein, protein-nucleic acid or protein-small molecule, not only helps to develop therapeutics or diagnostics techniques, but also provides important insights into cellular processes. Here we present a novel label-free and tether-free technology to analyze the affinity of biomolecular interactions based on the method Microscale Thermophoresis (MST). MST analyzes the directed movement of molecules in optically generated microscopic temperature gradients. This thermophoretic movement is determined by the entropy of the hydration shell around the molecules. Almost all interactions and biochemical processes relating to a change in size, charge and conformation of the molecules alters this hydration shell, and is thus detectable by MST. Here we show examples how MST can be used to quantify interactions in a label-free manner by using the intrinsic tryptophane fluorescence of membrane proteins and other interesting target molecules. In addition, examples are shown how MST can measure interactions with high selectivity in complex bioliquids like cell lysate or blood serum using a non-intrinsic source of fluorescence.
11:25 Exploring GPCR Desensitization Using Label-free Technology: Implications in Early Drug Discovery
Patricia McDonald, Associate Scientific Director, Translational Research Institute, Scripps Research Institute, Florida
G protein-coupled receptors (GPCRs) are by far the most diverse and therapeutically relevant class of receptors in the human genome. It is estimated that ~40% of all currently marketed drugs target GPCRs. Despite this success, emerging concepts in GPCR pharmacology such as ‘allosterism’ and ‘functional selectivity’ have led to significant efforts in identifying and optimizing receptor ligands with improved efficacy. Typically, ligand efficacy is defined by a ligand’s capacity to activate a single signaling pathway. However, it is now recognized that ligands can exhibit pluridimensional efficacies, a trait often overlooked when only considering a single cellular response. Activation of intracellular second messenger cascades, resulting from GPCR-ligand interactions, induce cytoskeletal rearrangement leading to changes in cell morphology. These rapid whole-cell responses can be measured using a label-free technology that measures changes in cellular impedance. Moreover, owing to the non-invasive nature of such technologies, in addition to detecting receptor activation; receptor ‘desensitization’, the process by which GPCR signaling is terminated, can also be detected and quantified. Desensitization of GPCR-mediated responses can arise from a number of different mechanisms depending on the receptor:ligand pair, cell-type, as well as sub-cellular localization of the receptor. The phenomenon of ligand-selective GPCR desensitization clearly has implications in the drug discovery process. Thus, a cell-based assay that monitors receptor desensitization regardless of the mechanism involved would greatly enhance the identification and optimization of GPCR leads. The use of emerging label-free technologies for the purpose of determining the propensity of various GPCR ligands to induce desensitization will be discussed.
11:50 Lunch on Your Own
Cell Based Assays
Moderator: Robert Hills, Senior Scientist, Integrated Systems Biology, Janssen Research and Development
1:30 Phenotypic Screen Assays for Drug Discovery
Wei Zheng, Group Leader, Therapeutics for Rare and Neglected Diseases, National Center for Advancing Translational Sciences, National Institutes of Health
Although the high throughput screening based on a known molecular target is currently a main approach for lead discovery, many of drug candidates identified through this process have encountered high failure rates in the late development stages including animal model tests and clinical trials. A recent review of failed drugs in clinical studies revealed that overall 31% of phase II and 61% of Phase III trials failed due to lack of human efficacy (data from 2007-2011, Arrowsmith, Nat Rev Drug Discov, 10: 87 and 10: 328, 2011). Questions have been raised to this drug discovery platform as the validated drug targets are reduced significantly over the time. Phenotypic screen, however, has recently emerged as an alternative approach for lead discovery that has a potential to interrogate human genome for new drug targets along with the discovery of new lead compounds. We have applied the phenotypic screen assays for several drug discovery projects involving the genetic diseases including Huntington disease, SMA, Niemann Pick Type C and beta-thalassemia. In addition, application of human cells differentiated from induced pluripotent stem cells (iPSCs) has recently emerged as a new tool for lead discovery that provides promising cell-based disease models for compound screens. We have used human neurons differentiated from iPSCs to assess the drug efficacy and cytotoxicity of a neuronal drug development program. The results and correlation between animal models and human neuronal cells will be presented and discussed.

Benefits:
• Examples of phenotypic screen assays
• Application of human iPSC derived cell-based disease model for compound screens
• Drug Repurposing screening
1:55 3D Cell-based Assays: Better in Vitro Biology for Drug De-risking
Jan Lichtenberg, CEO and Co-Founder, InSphero AG
To gain the full benefit of in-vitro cell biology, current cell monolayer models routinely used in drug discovery, have been further developed towards more physiologic systems. Although the advantages of organotypic 3-dimensional (3D) cell-culture models have been known for years, complex, low throughput production, and analytics impeded its industrial implementation.

New spherical microtissues can be produced in a high-throughput compatible hanging-drop culture system resulting in highly uniform spheroids in standard 96-well and 384-well formats. Microtissues can be generated from various cell sources and do not contain any artificial biomaterials such as hydrogels or scaffold materials.

The focus of the presentation will be on safety studies using primary rat and human liver 3D microtissues that reflect liver cell composition and include non-parenchymal cells. These 3D microtissues enable long-term toxicology assessments and the analysis of idiosyncratic toxicological effects in a scalable and automation-compatible assay process. The overview is complemented by examples of efficacy testing using single and multi-cell type tumor microtissue reflecting more closely the avascular tumor xenograft.

To be useful for drug de-risking, novel 3D cell-based models should seamlessly integrate into existing workflows. This requires a compatibility with existing biochemical read-outs, the possibility of histological analysis and adherence to standard formats. The presentation illustrates how these aspects can be addressed with scaffold-free, 3D microtissue spheroids to facilitate its adoption in daily screening routine.

Benefits of this presentation include examples for implementing 3D cell-based assays in commercial screening infrastructures and a complete picture starting from model production to assays and downstream analysis.
2:20 Optimizing Assay Methods for Interrogating 3D Spheroids
Terry Riss, Senior Product Specialist, Cell Health, Promega Corporation
Three dimensional (3D) cell culture systems provide a model to perform in vitro testing in an environment that exhibits cell-to-cell and cell-to-matrix interactions that are more physiologically relevant. Cells grown in 3D culture models have been shown to exhibit different signal transduction events, gene expression patterns and presence of biochemical markers, when compared to cells grown as monolayers. As the development and use of 3D models expands, there is a need to confirm the relevance of assay markers used as endpoints. There is also a need to confirm the performance of traditional cell-based assays designed for use with monolayer cultures, especially for the ability to lyse cells contained in large 3D structures or embedded in complex matrix components. The advantages of using a matrix-free hanging drop method for generating 3D microtissues of controlled size for verification of assay performance will be described. The performance of ATP detection as a marker of cell viability using 3D human liver microtissues will be shown. A collection of assays will be discussed for determining the cell stress pathway leading to cytotoxicity.

Benefits:
The hanging drop method:
- can generate 3D structures of controlled size that are reproducible for validating assay performance
- enables microtissue formation in the absence of added matrix components
- provides a single microtissue structure per sample well that avoids heterogeneity of other methods
The ATP cell viability assay:
- has adequate sensitivity for detection of small microtissues
- has the ability to effectively lyse cells in 3D microtissues
2:45 Organotypic Culture Array for High-Throughput Drug Screening
Paul Vulto, Senior Researcher, Leiden/Amsterdam Centre for Drug Research; CTO, Leiden University; MIMETAS
Recently, impressive progress has been made in mimicking organ functionality in a microfluidic space. However, upscaling of these techniques for massive parallel screening and their applicability in an industrial setting are issues that have been scarcely addressed.

We report a platform, based on phaseguide technology that enables arbitrary design and simple implementation of 3D cell and tissue culture models in a stratified approach. The tissue models are arrayed in a microtiter plate format. A continuous perfusion of nutrients and gas mimics the function of blood vessels in the human body. The system is equipped to facilitate arbitrary co-culturing and gradient conditions.

HepG2 hepatocytes and 4T1 murine breast cancer cells have been kept alive for three weeks with a 100% cell survival after two weeks. Cells proliferated progressively forming liver or tumor tissue. Liver tissue showed a dose dependent decrease in survival upon exposure to a panel of drugs that are known hepatotoxins. Cancer cell invasion and tumor formation was effectively demonstrated and quantified.

Phaseguides enable organotypic culture on a microtiterplate platform that is industry compatible and amenable for high-throughput screening without the use of any equipment other than standard pipetting aids. Continuous perfusion, mimicking blood flux, dramatically increases the life span of tissues and improves translatability of the model. The platform enables in-vitro drug efficacy and toxicity screening at higher predictability and higher throughput in order to reduce late stage attrition in drug development and develop better medicines for whole populations as well as for individual patients.

Benefits
• Create complex tissue models
• Using state-of-the art microfluidic techniques
• For early toxicity and efficacy screening
• By a passionate speaker with nice videos
3:10 Afternoon Networking Break
Novel Assay and Screening Technologies - Part I
Moderator: Terry Riss, Senior Product Specialist, Cell Health, Promega Corporation
3:40 Fragment Screening Strategies for Difficult or Unprecedented Targets - Leveraging SPR and NMR
Donald Huddler, Investigator, Biophysics Group – Computational and Structural Chemistry, GlaxoSmithKline
An increasing percentage of the contemporary drug discovery targets are novel or with limited precedence. The majority of these novel targets have little or no chemical matter in the literature. For the small percentage of truly novel targets that do have published chemical matter, many, upon careful examination, are revealed to be nuisance mechanism compounds of no utility. Consequently, the biophysics team is left with a challenging, novel target, with no validated compounds to develop and optimize direct binding screening assays. In our group, we have employed a general approach relying on saturation transfer difference (STD) NMR and SPR as well as a small, highly soluble, fragment library to quickly identify and validate reversibly binding compounds. Systematic application of this biophysics approach has yielded novel fragments for an array of non-traditional discovery targets.
4:05 You Want to Put Your Fragment Where? Hitting the Next (and really hard!) Generation of Targets with Fragments
Edward R. Zartler, Chief Scientific Officer, Quantum Tessera Consulting, LLC
The low hanging fruit has all been picked. The next generation of targets are more complex (and more difficult) which will require re-thinking how to screen them. These targets include unfolded (or intrinsically disordered), protein-protein interactions, multi-protein complexes, and a variety of targets no one would have touched a decade ago. NMR spectroscopy is a well-known screening and H2L follow-up technique that will have major impact in delivering hits on these next generation of techniques. This talk will discuss the obstacles these targets represent and ways that NMR (and other techniques) can be used to overcome them.
4:30 High Throughput Mechanotyping: Probing Cell Deformability Using Parallel Filtration
Amy Rowat, Assistant Professor, Department of Integrative Biology & Physiology, University of California, Los Angeles
Cell and nucleus mechanical properties are altered across a wide spectrum of physiological and disease contexts, ranging from genetic blood diseases to stem cell differentiation to cancer. We recently invented High Throughput Mechanical Screening (HTMS) instrumentation to simultaneously probe the deformability of >102 samples by subjecting them to external stresses, forcing them to deform through micron-scale pores, and counting the number of passaged cells. Importantly, HTMS provides a physiologically relevant screening assay to detect mechanical changes that have direct implications for the circulation and perfusion of cells through blood and tissues. This simple method is compatible with conventional multiwell plate formats, and requires only a pressure source and method for counting cells, such as flow cytometer or plate reader. Our preliminary results show that we can apply HTMS to probe the deformability of a variety of cell samples in parallel, including both adherent and suspension cells as well as the dose dependence of cytoskeleton-targeting drugs. With the ability to screen cell deformability across hundreds of samples, mechanical phenotype could be exploited as an effective label-free biomarker in fields ranging from cell mechanics to stem cell and cancer biology.
4:55 Screening for TRPV1 Positive Allosteric Modulators: A Route to New Analgesic Agents
Michael Iadarola, Chief, Neurobiology and Pain Therapeutics Section Laboratory of Sensory Biology, National Institutes of Health, National Institute of Dental and Craniofacial Research
New analgesic drugs are a major medical need. Opioids and non-steroidal anti-inflammatory drugs, the main current treatments, frequently are not well tolerated and are not always effective. Receptors and ion channels located on sensory neurons are attractive targets because they are unlikely to cause CNS side effects and The capsaicin or vanilloid receptor TRPV1 is one such molecule. This ion channel can be activated by noxious heat, low pH, lipid like endovanilloid compounds and is sensitized by inflammatory algesic compounds. Over-activation of the channel by TRPV1 agonists causes intracellular calcium toxicity and can functionally inactivate the pain-sensing nerve terminal producing analgesia in the area exposed to the agonist.

We have extended this nerve-terminal inactivation concept through the development of TRPV1 positive allosteric modulators (PAMs). TRPV1 PAMs are designed to act conditionally or in a state-dependent fashion only at sites of tissue damage or inflammation where TRPV1 is stimulated by endovanilloids or low pH.

Using a stably transfected cell line and a two-addition calcium fluorescence assay we screened the Molecular Libraries Small Molecule Repository for TRPV1 agonists and PAMs. A remarkable number of agonists were identified that were verified in a secondary dose-response screen. The PAMs were narrowed down to 4 lead compounds in addition to our initially identified compound (MRS1477). A variety of in vitro and in vivo testing and optimization has been performed which suggests that this is a new strategy for identifying potential analgesic therapeutic agents.

Benefit:
• Pain therapeutics, especially those directed at the peripheral nervous system
• Screening for ion channel activators and modulators
• Testing for analgesic agents
• Current development efforts for analgesic agents
5:20 Using the iCRO Model to Make a Repurposed Ion Channel Drug: The ChanRx Story
Rajesh (R.K.) Khosla, CEO, ChanRx Corp.
30 years ago, GBR 12909 (“vanoxerine”) started its journey from interesting molecule to Phase IIb human trials. Along the way, it has been through a myriad of in vitro, in vivo and human trials by a variety of companies. This presentation recounts the pathway from ChanTest’s discovery of vanoxerine’s ion channel profile 8 years ago, through pre-clinical testing, market targeting, Phase IIa human trial, market segmentation, strategic positioning, IP development, fund raising, assembly of the management team, SAB and SMB, and the imminent start of the Phase IIb dose ranging study for acute conversion of AF/AFL to sinus rhythm.

- Understand the commercial development process
- Understand the importance of market segmentation
- Understand how to structure the proper team
- Understand the difficulty of gaining outside validation
- Understand when to say “No” to requests from funders
5:45 Networking Reception and Poster Session
Day 1 Day 2 Day 3
Day 3 - Friday, October 26, 2012
7:30 Continental Breakfast
Novel Assay and Screening Technologies - Part II
Moderator: Wei Zheng, Group Leader, Therapeutics for Rare and Neglected Diseases, NCATS
8:00 Human Pluripotent Stem Cell Derived Hepatocytes and Applications in Drug Discovery
Petter Björquist, Senior Principal Scientist, Department Head, Cellectis Stem Cells
Human pluripotent stem cells (hPSC) hold an enormous promise for an unlimited source of specific cell types to be used as cell therapy in regenerative medicine. However, currently the most intense interest for hPSC is within direct industrial applications such as use as tools in drug discovery. Cell based in vitro assays with high human relevance are urgently needed for pre-clinical activities, spanning from target identification and validation, screening of compound efficacy, to drug metabolism and safety assessment studies. Hepatocytes are considered to be one of the most important cell types for these processes.

We have differentiated human embryonic stem cells (hESC) and human induced pluripotent stem cells (hiPSC) into hepatocyte-like cells by using a four-step differentiation protocol guiding the cells through discrete stages recapitulating liver development. The resulting cells morphologically closely resemble human hepatocytes and express hepatic markers on mRNA and protein levels. Additionally, the cells show hepatic functionality like albumin and urea production and CYP activity. The hPSC-derived hepatocyte-like cells are routinely produced in large quantities and made available in different multi-well formats.

We will show important aspects of industrial routine mass production of hPSC-derived hepatocyte-like cells. Examples of cell based screening assays will exemplify how these cells can be used in various industrial in vitro applications.

Benefits:
• Understanding of industrial aspects of the hPSC technology
• How hPSC can allow an unlimited source of functional cells
• Genetic diversity by using the hiPSC technology
• Assay development using hPSC-derived functional cells
• hES-HEP/hiPS-HEP, an unlimited source of hepatocytes for industrial applications
8:25 Targeting Membrane Associated Proteins Using Novel Template Directed Assembly (TDATM) Screening Technology
Kelvin Lam, Vice President, Blue Sky BioServices
Many plasma membrane-associated protein targets are found in multi-subunit complexes and the native membrane context determines the true biological activities. However, these drug targets are traditionally screened as isolated enzyme catalytic fragments that ignore the native environment. TDATM is a screening technology designed to solve this dilemma. TDATM reconstitutes a lipid environment for membrane-associated proteins and enables development of HTS assays that mimic physiological state of the native target. The TRK family are membrane-associated receptor tyrosine kinases (RTKs) activated by neurotrophins and implicated in neurodegeneration, pain, and cancer. TrkA was selected to validate TDATM technology. The assay was screened against a compound library, and we identified unique TrkA inhibitors using TDA. We concluded that TDATM is an enabling technology that preserves the proper polarity and topology for membrane-associated drug targets.

Benefits:
• Novel screening technology to target membrane-associated proteins
• Complementary technology to cell-based assays
• Identification of novel compounds
• Enabling technology to target membrane-associated targets
• Membrane-associated receptor tyrosine kinases (RTKs)
• Diverse small molecule compound libraries
8:50 Industry Leading Kinase Platform Innovation Drives Panel Efficiency in Lead Optimization
Jonathan Lippy, Senior Research Scientist II, Bristol-Myers Squibb
9:15 Application of Raman Measurement to ELISA
Neal Siegel, Chief Scientist, R&D, Sword Diagnostics, Inc.
Sword Diagnostics has developed an improved method for detection of peroxidase-linked Immunoassays based on Raman spectrophotometry. We have developed enzyme substrates that take advantage of the Raman light scattering effect to effectively improve accuracy, precision and sensitivity of Enzyme-linked Immuno-Sorbant Assays (ELISA). In many cases the improvements are immediate on the substitution of our detection chemistry into an existing assay. In some cases owing to the optimization of a particular set of reagents to the specific detection method some optimization on the assay developer’s part is required. Examples of these are provided in the body of the presentation.

Benefits:
• Novel application of Raman spectroscopy is shown to improve ELISA performance.
• Minimal intervention on the part of the Assay Developer is required.
• Improved detectability and quantitation can be realized for existing assays.
• Precision is improved across most of the dynamic range and especially at the lower end for most assays
9:40 Morning Networking Break
Advanced Methods in Drug Discovery
Moderator: Patricia McDonald, Associate Scientific Director, Translational Research Institute, Scripps Research Institute, Florida
10:10 Leveraging Combination High-throughput Screening to Identify Novel Drug Discovery Targets and Development Strategies
Glenn Short, Senior Director, Discovery Sciences, Zalicus, Inc.
Technological innovation has been both a boon and bust for drug development firms. While technology has increased the efficiency of drug discovery, emerging drugs designed to maximize on-target activity still suffer from high rates of attrition and sub-optimal clinical efficacy. The disconnect between clinical efficacy and drug selectivity stems, in part, from unknowns involving how drug targets are wired within cellular networks and how these networks respond to drug-challenge. To explore this knowledge-gap, we have leveraged our combination high-throughput screening (cHTS) platform in a number of discovery and development strategies that aim to improve efficacy. These strategies use combination chemogenomics to understand network responses to drug combinations and to map synergistic connectivity between targets. We and our partners are using this information not only to discover novel multi-target mechanisms, but also to lower the rates of attrition for targeted agents by minimizing toxicity, improving efficacy and identifying responsive patient subpopulations. Integration of these strategies into the canonical drug development paradigm will be discussed along with highlighted examples from our work using targeted drugs in the therapeutic areas of oncology and inflammation.

• Details cHTS approach and relevance to drug discovery and development
• Identifies drug development strategies in which cHTS can be used to improve translation
• Underscores the importance of understanding how the modulation of a drug target impacts cellular responses
• Highlights combination drug data in oncology and inflammation and its integration into traditional drug development strategy
10:35 High Throughput Behavioral Methods for Screening Novel Compounds Active in the Nervous and Muscle Systems
David Lowe, Head of Science, PsychoGenics Inc.
PsychoGenics has developed proprietary technologies for screening novel compound libraries using behavioral readouts at high throughput and scale. This phenotypic whole animal approach has the advantage that in vivo active compounds are identified early in the screening cascade, taking into account the composite interactions of bioavailability, brain penetration, receptor occupancy and other factors. Furthermore, structural motives with a poor side effect profile are identified based on overall behavioral parameters and can be eliminated as optimization candidates. This phenotypic screening approach utilizes computer based recording and analysis of rodent behavioral responses, pattern recognition, machine learning, bioinformatics and data mining. Signatures based on a large number of behavioral features, including resting and reactive behaviors, gait geometry and dynamics, social, circadian and cognitive behavior are identified for each compound, and can be compared to a reference data base of compounds with known mechanism and/or regulatory approval for one or more indications. Signatures can also be studied in both normal rodents and experimental transgenic, surgical or pharmacological models, giving a quantitative understanding of the ability of novel compounds to beneficially modify the phenotype in an experimental model, with again quantification of the compound’s side effect profile also being read out from the experiment. This approach has been successfully used in several drug discovery programs, for drug repurposing, as well as the detailed study of transgenic models of CNS and muscle disorders. The technology platform also shows great promise in the early detection of safety pharmacology signals. Several examples will be illustrated in this talk.
11:00 Discovering and Validating Drug Targets and Biomarkers using RNAi, High-Content Screening and Next Generation Sequencing
Donald Jackson, Senior Research Investigator II, Applied Genomics, Bristol-Myers Squibb Research & Development
RNA interference provides a fast and general method to assess the biological consequences of disrupting the expression of mammalian genes. Within the pharmaceutical industry RNAi has primarily been used to identify and characterize potential drug targets. However, RNAi is equally useful for identifying genes that modulate sensitivity to drugs, including response biomarkers and mechanisms of resistance. Screening highly multiplexed mixtures of RNAi reagents targeting over 10,000 genes followed by deconvolution with next-generation sequencing enable whole-genome scale screens in multiple models and conditions. Combining RNA interference with high-content screening enables screens with endpoints that could not be assayed otherwise, allows direct measurement of endogenous proteins, and provides additional information on mechanistic differences between hits as part of the primary screen results. We combine these approaches with additional genomic information from gene expression profiling, gene copy number analyses, and large-scale sequencing to deliver higher-quality targets and biomarkers with a greater likelihood of successful translation to the clinic.

Benefits:
• Use of RNA interference to understand mechanisms of drug resistance and sensitivity
• Combining RNAi with advanced assay platforms such as high-content screening and next-generation sequencing
• Highly multiplexed methods for rapid, large-scale RNAi screens
• Integration of RNAi results with other genomic data
11:25 Inhibitors of Nematode Detoxification Genes Identified from the NIH Molecular Libraries Collection
Siobhan Malany, Chemical Biology Team Leader, Conrad Prebys Chemical Genomics Center, Sanford Burham Medical Research Institute
Multidrug resistance in parasitic nematodes is a growing health and agricultural problem worldwide. The stress-inducible transcription factor SKN-1 regulates a significant number of detoxification genes in the model nematode Caenorhabditis elegans and is a promising target for the development of drugs that may be used in combination with current anthelmintics to inhibit drug resistance in parasitic species. We have established a 1536-well high-throughput whole organism-based assay to identify inhibitors of SKN-1 in C. elegans. Here, we report hit validation results from screening the NIH's Molecular Libraries compound collection (~360K) using acrylamide induced C. elegans in a dual fluorescent-based assay. Ratio of fluorescence intensities of the gene of interest and a housekeeping gene was determined in 1536-well format with an Envision reader. We also developed a counter screen assay to eliminate non-specific compounds. In summary, we have identified sub micro molar inhibitors specific to SKN-1. Structure-activity relationship studies have been initiated to identify a potent inhibitor of SKN-1 that may be used as a probe to better understand regulation of detoxification pathways and drug resistance in parasitic nematodes.
11:50 [Oral Presentations Submitted from Exemplary Abstracts]
Kinetics Based Early Drug Screening: A New Application for DNA-encoded Chemical Libraries
Nils Jakob Vest Hansen, CEO, Vipergen
A major limitation in drug discovery is the consideration of kinetic parameters such as ligand-target binding lifetimes in a high throughput early screen. We introduce a massively parallel screening method called binder trap enrichment (BTE) that addresses this critical problem. Here, a DNA-labelled target is exposed to a DNA-encoded chemical library (DEL) in solution. Then single protein molecules are trapped in emulsion droplets, with or without a ligand, during dissociation dominated kinetics - providing a snap shot of the solution - at controlled time points. Binding complexes are identified and counted by DNA ligation of the droplet contents, DNA amplification and deep DNA sequencing. With a very low false positive rate, true binding events are statistically differentiated over coincidental trapping of ligand and target. A 2-million compound DEL was screened against MAP kinase and carbonic anhydrase, yielding nanomolar affinity hits with the latter yielding an accurate ranking of inhibitors by their complex lifetime. Free compounds tested in binding assays confirmed the results validating BTE as the first high throughput method for screening large chemical libraries based on ligand-target binding lifetimes.
12:05 A Fluorescence Polarization Assay for the High-throughput Identification of Glutamate Carboxypeptidase II Inhibitors
Cyril Barinka, Institute of Biotechnology, Academy of Sciences of the Czech Republic
Glutamate carboxypeptidase II (GCPII) is a validated target for therapeutic and diagnostic interventions aimed at prostate cancer and neurologic disorders. To facilitate the identification of novel scaffolds inhibiting GCPII we have developed a high-throughput screening (HTS) assay based on fluorescence polarization (FP). To this end we prepared a fluorescence probe based on a urea-based inhibitory scaffold covalently linked to a Bodipy TMR fluorophore (TMRGlu) and implemented and optimized conditions suitable for HTS. Using known GCPII inhibitors, the FP assay was shown to be comparable to benchmark assays established in the field and as a proof-of-principle it was used to screen a 20,000-compound library of small molecules. The novel assay is robust, highly reproducible (Z’ = 0.82), inexpensive and suitable for automation, thus providing an excellent platform for HTS of small molecule libraries targeting GCPII.
12:20 Lunch Provided by GTC
1:15 Micropatterns for Structural Guidance of Cells
Myles Fennell, In Vitro Strategies, LLC
In vivo, cell behavior results from the integration of different signals obtained from adhesion molecules, paracrine and endocrine mediators and from mechano-structural guidance, a relationship that has been largely ignored in 2D culture systems. “Classical” in vitro 2D cellular assays are performed on unpatterned uniform adhesive substrates that can introduce considerable but unnoticed variability in cell function. 2D+ is a tool that provides 2D cultured cells with structural guidance. The guidance can be for individual cells or for groups of cells. Guidance can improve experimental reproducibility, sensitivity and provide a tool for understanding the relationship between structure and function in quantifying complex biological events. 2D+ restores the possibility for cells to adapt to their boundary conditions thus returning the opportunity to behave as mechano-sensing systems. Considering cells as unified systems, from their adhesive points down to their architecture, gene expression, and function, can improve the way cellular assays are performed and the quality of data generated. The 2D+ Platform provides structural guidance to cells cultured in vitro and thus the framework for expression of a more physiological phenotype. Innovative cellular assays will be described demonstrating the unique features of micropatterned cells showing new tools for quantification and biological analysis of complex cellular compartments.
1:40 Progressing Toxicology Testing Through the Use of Advanced Cell Models and HCA
Elizabeth P. Roquemore, Technology Manager, Cell Applications, Cell Technologies R&D, GE Healthcare Life Sciences
Development delays, black-box warnings and post-launch withdrawals of pharmaceuticals are driving the need for more rapid and predictive liability screening earlier in the development process. By simultaneously monitoring multiple toxicity indicators and phenotypic endpoints against the same cellular background, high content analysis (HCA) approaches can provide a more integrated and physiologically relevant means of evaluating toxicity than conventional in vitro tests performed in disparate assay systems. Moreover, recent advances in stem cell technologies have enabled robust and reliable production of relevant human cell models in quantities sufficient for high content screening. In this seminar, results from a multi-parametric live-cell study employing CytivaTM stem cell-derived cardiomyocytes will be presented to demonstrate the power of high content approaches in assessing the cardiotoxic potential of selective kinase inhibitors.
2:05 Ultra-high Throughput Screening Assays for Nuclear Hormone Receptors
Franck Madoux, Senior Scientist, The Scripps Research Institute
Nuclear Hormone Receptor (NHR) proteins form a class of ligand activated transcription factors that regulate the expression of downstream genes involved in a large breadth of biological functions, encompassing proliferation, differentiation, and homeostasis, both during development and in adult-hood. With 48 members described in humans, this protein family displays a wide variety of functions and transcriptional activities via the interaction with different co-activators and/or co-repressors. Unsurprisingly, this diversity, together with the significant function played by these key proteins, is reflected in the wide range of pathologies NHRs can be incriminated for, making this target class attractive for drug discovery. Since its creation, the Scripps Research Institute Molecular Screening Center successfully implemented a significant number of ultra-high throughput assays and screens to identify probes able to modulate directly or indirectly NHR activities in the miniaturized 1,536-well plate format. Through a series of practical and critical case-studies covering various targets and assay technologies, this presentation will present what ultimately became a broad, yet specialized platform to interrogate any NHR and allow not only to identify new lead series, but also to elucidate their mechanism of action, improve their potency, selectivity and bioavailability and, when applicable, assess their activity in vivo.
2:30 Translational Assays: A Bridge from the Bench to the Clinic
Robert Hills, Senior Scientist, Integrated Systems Biology, Janssen Research and Development
Many drug discovery assays rely on contrived systems or end points with little resemblance to what will actually be measured in the clinic. As a drug candidate progresses from initial HTS assay to in vitro and then into in vivo models, often different end points are measured in each respective assay. If one were to start with a set of target engagement markers gleaned from clinical data, then it would be possible to stream-line the discovery process by focusing on a set of consistent markers that translate across each step. In all cases, label free technologies are employed, with LC/MS/MS being one of the more useful and powerful techniques.

• What makes a good translational assay?
• What technologies are amenable to this assay format?
• What are the potential pitfalls?
2:55 Conference Concludes
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