Institute for Scientific Exchange, Inc. Presents:

 

5^th International Conference on Early Toxicity Screening:

Strategies and Approaches for Toxicity Screening in Drug Discovery and Development

February 21-23, 2005

San Diego, CA, USA

Symposium Venue:  San Diego Mission Valley Marriott

 

Featuring experts from the following institutions: Merck & Co., Inc; Millennium Pharmaceuticals Inc.; Institute for In Vitro Sciences, Inc.; National Cancer Institute; University of California; SRI International; NIEHS; ILS, Inc.; Duke University Medical Center; Texas A&M University; Phylonix Pharmaceuticals, Inc.; Multi Channel Systems MCS GmbH; Novartis Institutes for Biomedical Research; RegeneMed, Inc.; Advanced Pharmaceutical Sciences, Inc.; Pfizer - Groton Labs; Puracyp, Inc.; Promega Corporation; BioSource; Cambrex Bio Science; Allergan; CeeTox, Inc.

 

Featured Exhibitors:

 

 

 

 

 

 

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Monday, February 21, 2005

 

 

8:00 AM – 9:00 AM – REGISTRATION

 

8:00 AM – 5:00 PM – EXHIBITS

 

8:50 AM – 9:00 AM – Exhibitor Presentation – Cambrex Bio Science Walkersville, Inc.

 

9:00 AM – 9:30 AM

Screening Strategies to Reduce Reactive Metabolite Formation in Drug Discovery (Weichao Chen, Merck & Co., Inc. San Diego, CA) Toxicity of drug candidates is one of the major problems facing the pharmaceutical industry. Inadequate detoxification of reactive drug metabolites formed through bioactivation has been implicated as a pathogenic mechanism for tissue necrosis, carcinogenicity and immune-mediated toxicity.  Thus, implementing screens to weed out compounds with potential toxic metabolites in early drug discovery may enable all drug candidates entering development to achieve a higher survival rate.  Screening methods to identify candidates with the potential to produce reactive metabolites will be presented.

 

9:30 AM – 10:00 AM

In Vitro and In Silico Screening Strategies in Hit-to-Lead (H2L) and Early Lead Optimization (LO) With Good Predictivity for Development Limiting Side Effects and Toxicity (Scott A. Barros, Millennium Pharmaceuticals Inc.; Cambridge, MA) Further improvement in efficiency from a toxicology and safety pharmacology perspective in pharma R&D lies in leveraging “best in class” technology and early integration with medicinal chemistry and pharmacology activities during H2L and early lead optimization. Screening assays must be reasonably high through put and low in test agent consumption. Our in silico and in vitro testing paradigm for toxicity and side effects has well defined predictivity for human response and for development limiting barriers focusing on QT interval prolongation, liver toxicity, and genetic toxicity. The strategy and tactics as well as supportive data will be presented in detail.

 

10:00 AM – 10:30 AM

Validation & Regulatory Acceptance of In Vitro Toxicity Assays (John W. Harbell, Institute for In Vitro Sciences, Inc.; Gaithersburg, MD) This session will focus on the design and validation of in vitro bioassays intended for use in the regulated environment.  Bioassay are designed around four components; a specific cell or tissue, defined exposure conditions, characterized endpoint measures of the biological effect, and a predetermined relationship between the endpoint values and the effect being predicted (potency or toxicity). The assay’s predictive capacity is dependent on maintaining consistency in these parameters from assay validation through its routine use for regulatory submissions.

 

10:30 AM – 10:45 AM – BREAK

 

10:45 AM – 10:55 AM – Exhibitor Presentation – APSciences, Inc.

 

10:55 AM – 11:15 AM – PANEL DISCUSSION: Session I

 

 

11:15 AM – 11:45 AM

Overview:  Screening for Organ Toxicity for Anticancer Drugs (Joseph E. Tomaszewski, National Cancer Institute; Rockville, MD) Anticancer drugs are known to have toxicity towards normal organs, especially to proliferating cells such as bone marrow and intestinal epithelium.  Other toxicities include hepatotoxicity, nephrotoxicity, and neurotoxicity.  The experience in NCI on organ toxicity will be summarized, with emphasis on species-species differences.

 

11:45 AM – 12:15 PM

Development of In Vitro Bioassays to Predict Human Response (Eugene Elmore, University of California, Chao Family Comprehensive Cancer;

Irvine, CA) The prediction of clinical efficacy and adverse events using in vitro response data has been difficult due to the lack of direct evidence that in vitro assays were responsive at concentrations achievable in humans.  Our laboratory has developed a variety of in vitro assays that utilize human cells to identify efficacy for cancer prevention as well as tissue specific toxicity at clinically relevant concentrations.  Data will be presented on our approach to assay development and the correlation to human data. 

 

12:15 PM – 1:30 PM – LUNCH

 

1:30 PM – 2:00 PM 

New Applications for Precision-Cut Slices from Liver and Lung (Holger P. Behrsing, SRI International; Menlo Park, CA) Recent improvements in technique have considerably extended the survival time of liver and lung precision-cut slices in roller cultures and thereby their usefulness as toxicology models.  By coupling these improved systems with histology, immunohistochemistry, and traditional clinical chemistry biometrics, it is shown that toxicants induce histological and biochemical changes remarkably similar to those they induce in vivo.   This talk will present data supporting this conclusion and examples of applications not previously reported for these systems.

 

2:00 PM – 2:30 PM

Organ Slices for In Vitro Toxicology (Alison Vickers, Allergan, Inc., Irvine, CA) Tissue slices, an in vitro model representing the multi-cellular and functional features of in vivo tissues, is a promising model for characterizing mechanisms of drug induced organ injury and for defining species susceptibilities to organ injury. The integration of gene expression analysis, linked with assessments of cell function and morphology, contribute to our understanding of key cellular pathways leading to organ injury. However, it is the application of human tissue that enhances our ability to focus on the clinical relevance of a potential outcome, and for identifying clinically relevant biomarkers.

 

2:30 PM – 3:00 PM

International Validation of a CFU-GM Test for Predicting Neutropenia in Man  (Ralph E Parchment, SciTech Development LLC; Grosse Pointe, MI) The international validation of a CFU-GM test for predicting acute doses of chemicals that will cause severe, reversible neutropenia in man has led to the discovery of important general principles for predicting organ system toxicities using in vitro assays.  Importantly, the predictive IC (effect level) in an in vitro test appears to be unique to each organ system, and depends on in vivo tissue reserve and the tissue-specific relationship between loss of function and clinical manifestation of toxicity. It was also realized that the in vitro exposure-effect relationship does not reveal hazard classification per se, but instead establishes risk of toxicity in a particular organ system as a function of dose.  To be useful in practice, the in vitro endpoints must be relatable to formalized classifications of clinical toxicity used by regulatory agencies.  Thus, predicting clinical toxicity profiles and safe exposure levels in man without using animal models will require a battery of in vitro assays that each predicts the safe exposure level of an innovator chemical for a particular organ system’s clinical function (“mechanism-naïve” assessments).  Since these in vitro systems assess the adverse effects of toxic chemicals on the actual target tissue affected in vivo, and provide information about clinical outcome of different exposure levels, it is expected that in vitro human safety pharmacology studies will ultimately be more accurate at predicting safe human exposure levels than animal models.”

 

3:00 PM – 3:15 PM – BREAK

 

3:15 PM – 3:45 PM

Systematic Detection of Hemotoxicity to Stem and Progenitor Cells Using the HALO Platform (Ivan N. Rich, HemoGenix, Inc.; Colorado Springs, CO) HALO (Hemotoxicity Assays via Luminescence Output) is a multifunctional in vitro platform specifically designed to examine the effects of virtually any compound on stem and progenitor cells of the hematopoietic system. Validation of this proliferation assay has used 11 reference compounds from the Registry of Cytotoxicity Database to examine their cellular effects, IC50/IC90 values and rank the compounds in order of their toxicity on 7 different cell populations derived from human and mouse bone marrow.

 

3:45 PM – 4:15 PM – PANEL DISCUSSION – Session II

 

 

4:15 PM – 4:45 PM

Models to Examine the Peripheral Nervous System as a Target Site of Damage (Jean Harry, NIEHS; Research Triangle Park, NC) Adverse side effects of pharmaceutical agents can include a disruption in the normal functioning of the peripheral nervous system.  Quite often these effects require a period of time to manifest yet can become debilitating to the patient.  This presentation will provide a brief general overview of the peripheral nervous system, examples of compounds that selectively effect components of this system, in vivo screening methods, and in vitro models that have been developed to identify and understand mechanisms underlying such alterations.

 

4:45 PM – 5:15 PM

High-Content Screens for Neurodegeneration and Neurotoxicity Based on Brain Tissue Explants (Donald C. Lo, Center for Drug Discovery, Duke University Medical Center; Durham, NC) Cell-free, cell line, and primary culture based screens often do not adequately predict either therapeutic benefit or neurotoxicity in the intact brain.  Such discordances arise, at least in part, from the lack of normal tissue context for complex organ systems such as the brain.  We have thus developed disease-specific, neural screening assays incorporating intact and living brain slice explants, and show their use in drug and drug target discovery, and in determining genetic predisposition to neurotoxicity in Parkinson's disease.

 

END OF DAY

 

 Tuesday, February 22, 2005

 

 

8:00 AM – 9:00 AM – REGISTRATION

 

8:00 AM – 5:00 PM – EXHIBITS

 

8:50 AM – 9:00 AM – Exhibitor Presentation - StemCell Technologies

 

9:00 AM – 9:30 AM

Concepts of In Vitro Neurotoxicology: Applications to Drug Discovery (Evelyn Tiffany-Castiglioni, Texas A&M University; College Station, TX) This presentation will present concepts of in vitro neurotoxicology that should facilitate early toxicity screening for drug discovery.  Emphasis will be on improved methods and systems for in vitro neurotoxicity testing, and their relevance to in vivo central nervous system toxicity.  Pharmaceuticals will be compared with well-known neurotoxicants.  The topics addressed will be: utility of in vitro systems, neurotoxicants and their cellular targets, the paradigm of accumulated damage, interacting biological facets of in vitro neurotoxicology, and extrapolation of benchmark doses in vitro for neurotoxic endpoints.

 

9:30 AM – 10:00 AM

Zebrafish Assays for Drug Effects on the Nervous System (Chaoyong Ma, Phylonix Pharmaceuticals, Inc.; Cambridge, MA)  Current success rate to market for nervous system drugs is the lowest among major therapeutic areas, and numerous approved drugs are associated with neurotoxicity. The zebrafish model can help solve these problems by enabling rapid and thorough assessment of drug effects on the nervous system during early stages of drug development. We have generated zebrafish models of nervous system diseases by gene knockdown and neurotoxins. We also showed that compounds that induce neurotoxicity in humans caused similar neurotoxicity in zebrafish.

 

10:00 AM – 10:30 AM – PANEL DISCUSSION – Session III

 

10:30 AM – 10:45 AM – BREAK

 

 

10:45 AM – 11:15 AM

Automated Repolarization Assays for Early Cardiac Toxicity Screening (Thomas Meyer, Karl-Heinz Boven, and Christine Leisgen, Multi Channel Systems MCS GmbH; Reutlingen, GERMANY) Extracellularly recorded cardiac field potentials allow monitoring drug effects on cardiac action potentials. With this method, cardiac ionic channels can be studied in their natural environment. Spontaneously beating cardiomyocytes allow the detection of second-messenger mediated effects and arrhythmia. The automated QT-Screen system uses 96-well-plates for achieving a medium to high throughput.  Dose response relationships have been analyzed for E-4031, Quinidine, Sotalol, Verapamil and IKATP modulators Glibenklamide, H1098, and Rilmakalim. The data is in line with published data from repolarization assays.

 

11:15 AM – 11:45 AM

Implementation of a Novel Early Toxicology Strategy for Genotoxicity and Cardiosafety Compound Profiling in the Drug Discovery Process (Claudia McGinnis, Discovery Technologies, Novartis Institutes for Biomedical Research, Cambridge, MA) Comprehensive Early Toxicology Profiling of lead compounds is increasingly being considered as a novel strategy to reduce attrition rates in development. The principal safety concerns are usually in the area of genetic toxicology and cardiosafety for which standard experimental procedures are well established in low throughput format. The challenge is to identify and evaluate predictive, higher throughput in vitro and in vivo assays and technologies, and to implement them into an automated screening environment.  An overview of novel, high throughput genetic toxicology and cardiosafety screening scenarios will be presented. Predictability, compound consumption, and ease of implementation of these assays will be compared to next level, traditional toxicity tests. Results will be presented from a number of external validation and in-house collaborative studies and examples will be shown how to fit the concept of early toxicology profiling into the drug discovery process.

 

11:45 AM – 12:15 PM

The Use of Genetic Toxicity Assays in Drug Decision Making (Raymond R. Tice, ILS, Inc.; Research Triangle Park, NC) Before drugs are made publicly available, an assessment must be made of the risk for adverse health consequences, including a genotoxicity evaluation (i.e., the ability to damage/mutate DNA). Current regulatory genetic toxicology testing consists of a battery of complementary in vitro/ in vivo assays evaluating various endpoints (mutagenicity, chromosomal aberrations, micronuclei, and/or DNA damage).  Some genotoxicity tests are used for mechanistic studies. The basis for these tests,

their limitations and advantages, and how genotoxicity data are interpreted will be reviewed.

 

12:15 PM – 12:30 PM – PANEL DISCUSSION: Session IV

 

12:30 PM – 2:00 PM – LUNCH

 

 

2:00 PM – 2:30 PM

3D Tissue Engineered Human Liver for ADMET (Dawn R. Applegate, RegeneMed, Inc.; La Jolla, CA)

Currently available ADMET screening methodologies express transient activity and are not fully predictive of human function. As a result, the majority of drug candidates fail clinical testing. Tissue engineered human and animal liver provide organotypic cultures that recreate the in vivo microenvironment, containing all of the cells of native liver in an extracellular matrix the cells themselves create. These cultures afford long-term maintenance of tissue-specific function in vitro as well as provide predictive responses to model chemical and drug toxins.

 

2:30 PM – 3:00 PM

Early Toxicity Testing for Human Organ-Specific Toxicity:  From Hepatocytes to IdMOC (Albert P. Li; Advanced Pharmaceutical Sciences, Inc.; Baltimore, MD) Because of species-differences in drug toxicity, animal models do not always predict human drug toxicity.  The use of primary cells from human organs allows the generation of data relevant to human organ toxic effects of drugs and drug candidates.  For instance, human hepatocytes and renal tubule cells can be used for the evaluation of the adverse effects of a chemical on liver or kidney, respectively.  A further advancement on the application of primary cells is the Integrated Discrete Multiple Organ culture (IdMOC) system which allows multiple cell types to be cultured physically separated but interconnected by a common medium, akin to an animal (human) in vivo, where multiple organs are connected by the circulating blood.  The IdMOC allows the evaluation of the toxicity of a chemical on multiple organs, with multiple organ metabolism which is lacking in single cell-type systems.

 

3:00 PM – 3:30 PM

Applications of Cell-based Imaging Technologies in Toxicity Screening in Drug Discovery and Development (Jinghai (Jim) Xu, Pfizer - Groton Labs; Groton, CT) Traditionally, in vivo histopathological findings have been used as part of standard readouts for safety assessment of new chemical entities (NCEs).  The in vitro correlate of these readouts would be cell-based image analysis.  This presentation will present an overview of the current state of the art using cell-based imaging to predict human toxicity, esp. targeting mechanisms of necrosis, steatosis, phospholipidosis, micronucleus, mitochondria dysfunction, oxidative stress, and drug interactions.  The applications of automated image analysis in assays predicting phospholipidosis and micronucleus will be highlighted.  It is anticipated that one or more of these technologies can be used to identify NCEs with higher probability of safety in earlier phases of drug discovery and development.

 

3:30 PM – 3:45 PM – BREAK

 

3:45 PM – 4:15 PM

A Simultaneous Assessment of CYP3A4 Metabolism and Induction in the DPX-2 Cell Line (Scott Allen, Puracyp, Inc.; Carlsbad, CA) Exposure to certain xenochemicals can alter the catalytic activity of the major drug-metabolizing enzyme, CYP3A4, either by enhancing expression of this P450 or by inhibiting its activity.  A simplified and reliable tool for detecting the ability of candidate drugs to alter CYP3A4 levels or inhibit its catalytic activity was developed by construction of a stable cell line.  Treatment of these stable transformants (DPX2) with various known CYP3A4 inducers produced increases in luciferase expression that correlated to CYP3A4 mRNA levels.  DPX2 cells were also used to identify inhibitors of CYP3A4 catalytic activity and assess toxicity of several agents.  Collectively, DPX2 cells can be used to recognize xenobiotics that are toxic to mammalian cells and simultaneously identify agents that induce or inhibit CYP3A4 enzyme activity.

 

4:15 PM – 4:45 PM – PANEL DISCUSSION: Session V

 

END OF DAY 

Wednesday, February 23, 2005

 

 

7:00 AM – 8:00 AM – REGISTRATION

 

7:00 AM – 12:00 PM – EXHIBITS

 

8:00 AM – 8:30 AM

Multiplexing In Vitro Cytotoxicity Assays (Terry Riss, Promega Corporation; Madison, WI) Recent advances in cell-based assay chemistries have made it possible to record multiplex data in a high throughput format using standard plate readers without the need to perform microscopic imaging of individual cells. Development of non-destructive endpoints for cell-based assays has expanded the possible combinations for multiplexing; however, for homogeneous multiplexing to be successful, the assay chemistries must be compatible. We have developed methods to combine various luminescent and fluorescent cell-based assays in a homogeneous format. We have recorded luciferase reporter gene assays in real time in living cells followed by measuring cell viability or caspase activity. Measuring the number of viable cells at the end of a treatment period is useful to distinguish between a specific down regulation of a reporter gene or non-specific cytotoxicity. Multiplexed cell viability data also can serve as an internal control to correct for variability in seeding density and differential growth of cells resulting in improvements of the reliability of data. We will present the results of ongoing efforts to investigate the compatibility of different assay chemistries and detection methods for developing multiplexed homogeneous cell-based assays.

 

8:30 AM – 9:00 AM

A Review of Methods for Quantifying Cytotoxicity and Apoptosis (Mary Michael Brodey, BioSource; Camarillo, CA) The drug discovery process can be facilitated by studies that quantify cytotoxicity and apoptosis. Quantification of cytotoxicity can be accomplished by several methods. Cells can be stained using compounds that are specifically taken up by viable cells (e.g., CFDA, Neutral Red, or Europium Titriplex V) or using compounds that are specifically excluded from viable cells (e.g., propidium iodide or Hoechst 33342). Cytotoxicity can be assessed by monitoring release of LDH or  ⁵¹Cr into culture media. Cytotoxicity can also be assessed by monitoring cellular redox state using alamarBlue^TM or tetrazolium salts. Methods for monitoring apoptosis rely on the detection of physiological processes that accompany this cell function. The changes in the plasma membrane that occur during apoptosis can be detected and quantified by staining cells with Annexin V-FITC. The DNA fragmentation that occurs during apoptosis can be detection and quantified by analyzing chromosomal DNA by gel electrophoresis, as well as by measuring bromodeoxyuridine incorporation into broken DNA ends. The caspase cleavage and activation observed in apoptotic cells can be monitored by Western blotting and caspase enzyme activity determinations that use labeled synthetic peptide substrates.  A review of these methods for detecting cytotoxicity and apoptosis will be provided.

 

9:00 AM – 9:30 AM

Rapid Luminescent Assays for Cytotoxicity (Alex Batchelor, Susan Gill, Anthony Pitt; Cambrex Bio Science; Nottingham, UK) Luciferase/luciferin assays have been used in drug discovery for many years to determine cell viability, proliferation and cytotoxicity. The sensitivity of luciferase enables the detection of ATP from as few as 10 cells, resulting in assays that miniaturise easily with repeatable IC50s. More recently, other luminescent cytotoxicity assays have been developed, measuring adenylate kinase for cytolysis, or caspase activity for apoptosis. We will present a review of available methods for determining cytotoxicity in HTS formats. We aim to highlight the differences between the various luminescent assays and show comparisons with non-luminescent assays. Key considerations are what cellular process is measured,  at what point in the cell death process the measurement is taken, how assay sensitivity affects IC50 and how robustness of the assay technology reduces the chances of false positive and false negative results.

 

9:30 AM – 9:45 AM – BREAK

 

9:45 AM – 10:15 AM

In vitro Toxicity Screens That Focus on Key Cellular Functions Provide Essential Information Early in Drug Development (James M. McKim, CeeTox, Inc.; Kalamazoo. MI) This presentation will focus on a new approach to in vitro toxicity screening that is based on multiple endpoint analysis of key biochemical functions that are essential for cell health.  The system of toxicity screening combines multiple endpoints, dose-response, time, and a database of non-proprietary drugs and chemicals to provide information on the adverse properties associated with new chemical entities.  The biochemical data obtained provides a high degree of in vivo predictability.  The data obtained can be used by researchers to improve the safety profiles of potential drug candidates at a time when modifications to the chemical structure can still be made.  The end result is more data that can be used to make better decisions earlier in the development pipeline. 

 

10:15 AM – 10:45 AM – PANEL DISCUSSION: Session VI

 

END OF CONFERENCE

 

 

 

POSTER PRESENTATIONS:

 

Poster Presentations are always encouraged.  Please submit your poster abstract for approval by the organizing board by January 30^th.  Poster size should be no larger than 4 feet high by 7 feet long.  Abstracts of posters will be included in the participant binder and in the ISE website.  There is no formal poster presentation scheduled.  All posters will remain displayed throughout the conference.  Please be prepared to display your poster during registration on Sunday February 20^th or before the first session begins on Monday, February 21^st.  Poster presenters will have ample time for discussion during breaks and the Welcome reception. Submit posters abstracts for approval to Nola Mahaney, VP, Operations; ISE, Inc.; 5707 Calverton Street, Suite 2C; Baltimore, MD 21228 or fax at (410) 869-9560 or email file attachment to nola@isciencex.com.  Approved poster presenters are responsible for completing a conference attendance registration form and payment of fee (visit www.isciencex.com/register.htm) and for the shipping of the poster itself.  Please contact Nola Mahaney for any questions or concerns.  Please refer to “Travel Information” for hotel address and shipping information.

 

Travel Information

The Symposium venue, the San Diego Marriott Mission Valley is conveniently located at Interstate 8. Exit Qualcomm Way to Rio San Diego Drive, just ½ mile west of Qualcomm Stadium. Located minutes from Qualcomm stadium, San Diego Zoo, Sea World©, and the San Diego Mission. A number of restaurants, shops and services are also nearby.  Please contact the hotel for further information.

 

Hotel Information

A limited number of rooms have been reserved at The San Diego Marriott Mission Valley, 8757 Rio San Diego Drive; San Diego, CA, 92108 USA, at a reduced conference rate of US $129.00 per night (plus applicable taxes-single occupancy).  Please make your hotel reservation directly by telephone at (800) 842-5329, (619) 692-3800, fax (619) 692-0769 or www.marriott.com.  Please ask for “ETS-2005” block for ISE, Inc.

 

Payment

Payment may be made by check or credit card.  Checks should be made in US $, payable to Institute for Scientific Exchange, Inc.  Mail to: ISE, Inc., 5707 Calverton Street, Suite 2C, Baltimore, MD 21228, USA

 

Cancellation Policy

All cancellations are subjected to a $250.00 cancellation fee. Longer than 30 days, 100% refund (less cancellation fee). Less than 30 days, no refund but registration may be transferred to another person.  All refund requests must be in writing.  All refunds will be issued after the meeting has occurred. No refunds requests will be accepted after January 21, 2005. Please submit cancellation and refund requests including transferring of registration to:

 

Fax:  410-869-9560

E-mail: nola@isciencex.com

Deadline: January 21, 2005

 

Email to nola@isciencex.com or mail/fax completed form with remittance to: ISE, Inc. 5707 Calverton Street, Suite 2C, Baltimore, MD 21228, USA; FAX No:  (410) 869-9560.  Payment may be made by check in US$, payable to Institute for Scientific Exchange, Inc. or by credit card.

 

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Payment Method:  Check, Credit Card (VISA, MASTER CARD, AMERICAN EXPRESS)

 

ETS-2005 Registration:     US $1500.00________

Transporters-2005:             US $1500.00________

 

SPECIAL DISCOUNT-BOTH EVENTS US$2000.00

 

Exhibitors:                             US $2000.00________

 

Academic/Government participants will receive a 50% discount. 

 

Contact Nola Mahaney for Exhibitor or Sponsorship Opportunities at nola@isciencex.com, or phone (410) 869-9166); or visit www.isciencex.com/exhibitors .

 

REGISTER NOW