Wednesday, December 12, 2001
Registration & Exhibit Set-up: 5:00 7:00 PM
Thursday, December
13, 2001
Symposium I In Silico Toxicology
Chairs: Alan G. E. Wilson, Pharmacia Corp.
Dale E. Johnson, DDPlatform, LLC
9:00 10:00 AM
Continental Breakfast and Registration
10:00
10:45 AM
Keynote Speaker
Role of In Silico Modeling and Prediction in
Drug Safety Evaluation - Past, Present and Future (Alan
G. E. Wilson, Pharmacia Corporation; St. Louis, MO)
10:45 11:30 AM
The
Use of Chemoinformatics in In Silico
Toxicology Evaluation. (Dale E. Johnson,
Eos Biotechnology, S. San Francisco, CA) Correlating chemical
structure features statistically with toxicological endpoints requires a
complex chemoinformatics approach. For the design
of chemical libraries or optimization of lead structures, the use of a
medicinal chemistry building block approach is an effective model for
chemoinformatics. The application of this chemoinformatics approach to in
silico toxicology will be discussed.
11:30 11:45 AM - BREAK
11:45
AM 12:30 PM
Toxicity Evaluation Using Statistical Analysis and
Visualization. (Tom
Downey, Partek Incorporated; St. Charles, MO) Early
assessment of toxicological effects involves a variety of technologies
including high throughput screening (HTS), structure activity relationships
(SAR), and measurement of gene expression. In order to extract meaningful
discoveries, a systematic approach of statistical data analysis and
visualization is needed. This talk will explain concepts of experimental
design, exploratory analysis and inferential statistics (explained in layman's
terms) as they apply to the above technologies.
12:30
1:15 PM
In Silico Approaches
for the Prediction of Toxicity and Metabolic Fate. (Anita White, Pharmacia Corp., St. Louis, MO). An integrated
approach using multiple commercial in silico platforms has been developed and
applied towards Pharmacia proprietary chemicals.
Details of this integrated approach and the statistical analysis of its
performance will be presented.
1:15 2:30 PM
- LUNCH BREAK
2:30 3:15 PM
In Silico Toxicology: Pfizer Perspectives (Nigel Greene,
Pfizer Global Research and Development; Groton Laboratories; Groton, CT) The
approaches and experience in the application of in silico toxicology in Pfizer
will be discussed.
3:15 4:00 PM Round Table discussion
END OF DAY
6:00 PM 8:00 PM Welcome Reception (cocktails
and hors doeuvres, casual attire, not mandatory)
Friday, December 14, 2001
Symposium
II: In Vitro Approaches
Chairs: Albert P. Li, In Vitro Technologies and
Michael P. Carver, Wyeth Ayerst Research
8:00
9:00 AM
Continental
Breakfast and Registration
9:00 9:45 AM
Screening for Hepatotoxicity with Cryopreserved Human
Hepatocytes: In
Vitro Model for Phase I Clinical Trial.
(Albert P. Li, InVitro
Technologies, Inc., Baltimore, MD) Multiple endpoints
for toxicity have been developed with cryopreserved human hepatocytes and
applied to model hepatotoxicants. In additional to high throughput
screening using a single endpoint, we have also developed an approach with
which a single hepatotoxicant can be evaluated with hepatocytes from multiple
donors, similar to the testing of multiple subjects in clinical trials.
9:45
10:30 AM
A Method for the Quantification of Reactive Metabolite
Formation from Toxic Drugs in Human Hepatocytes. (Neil Hartman, CDER, USFDA, Laurel, MD) Reactive
metabolite formation is an important factor in the understanding of
idiosyncratic drug toxicity. An HPLC method has been developed in our
laboratory using radio labeled glutathione for the quantification of the
formation of drug-glutathione conjugates. This method may provide useful
information for the estimation of toxic potential, especially towards
idiosyncratic drug toxicity
10:30 10:45 AM BREAK
10:45
11:30 AM
In Vitro Nephrotoxicity Evaluation Using Primary Human Kidney Cells (Scott Lloyd, In Vitro Technologies, Inc., Baltimore, MD) Besides
hepatoxicity, nephrotoxicity is a major toxicological outcome of adverse drug
effects. Primary cultures of human kidney tubules cells are used in our
laboratory as an experimental model for nephrotoxicity, using ATP content, MTT
metabolism, neutral red uptake, and alamarbluemetabolism as endpoints.
The response of both primary cultures and cultures of cryopreserved human
kidney to model nephrotoxicants will be discussed.
11:30
AM 12:15 PM
In
Vitro Screening
Assay for Bone Marrow Cell Toxicity. (Joe Tomaszewski, National Cancer
Institute, Bethesda, MD) Bone marrow toxicity is a major concern
for chemotherapeutic agents. In NCI, in vitro bone marrow cell cultures
from rodents, dogs, and humans are used for the screening of potential
toxicity. Strength and challenges of this important in vitro screening
assay will be discussed
12:15 1:30 PM
- LUNCH BREAK
1:30
2:15 PM
Predictive Toxicity Using Multiple Gene Expression Platforms (Michael Orr, GeneLogic, Inc.; Gaithersburg, MD) Open and closed system gene expression technologies; Multiple species approach; Building reference database for in vivo and in vitro use; Entire genome assayed for predictive qualities; Prediction of toxicity across compound classes; Discrimination of individual compounds and classes.
2:15
3:00 PM
n Vitro Evaluation of Eye and Skin Toxicity. (John
Harbell, Institute for In Vitro Sciences, Inc.) Various in
vitro methods are currently available for assessing potential eye and skin
irritation. Some of the most promising are the reconstructed human tissue
models, which can be used both for safety and efficacy studies. A review of
these models and the progress of international validation studies in this area
will be discussed.
3:00
3:15 PM - BREAK
3:15 4:00 PM
In Vitro ADMET Profiling in Early
Drug Discovery. (O. Helen
Chan, ArQule, Inc, Woburn, MA) A Parallel Track Drug Discovery
process involving simultaneous generation and evaluation of experimental data
for ADME, toxicity, selectivity, solubility, and potency to develop
multi-dimensional structure-activity relationships that guide chemical
synthesis will be presented.
4:00 4:45 PM
- Round Table Discussion
END OF DAY
Saturday,, December 15, 2001
Symposium
III: HTS Approaches
Chairs:
John Watson, Promega Corp.
Uday Saxena, Reddy US Therapeutics
8:00 9:00 AM
Continental Breakfast and Registration
9:00 9:45 AM
High Throughput Toxicity Assays for Drug Candidate
Prioritization: Predictive Value, Simplicity and Sensitivity. (John Watson, Promega Corporation, Madison, WI)
The purpose of this talk is to discuss how and why toxicity
assays are moving towards earlier stages in the drug discovery process. An
emphasis
will be placed on methods that have the best correlation with human toxicity
outcome data. Two commercially available methods that assay cell viability
via mitochondrial metabolites will be described with an emphasis on how the
assays can be used in high throughput applications.
9:45 10:30 AM
High Throughput ADME-Tox screening with the Acumen Explorer. (Adrian Dawkes, Acumen Bioscience Ltd., a TTP
Group Co., Herts, UK) The
acumen explorer provides parametric measurements for fluorescent cell based
assays in high throughput and high content modes. Data will be presented
showing the applicability of the instrument in ADME-tox screens across a range
of cell and tissue types.
10:30 10:45 BREAK
10:45 11:30 AM
Oxidative DNA Damage is Involved in Many Pathological Processes,
Including Drug and Chemical Toxicity. (Jean-Paul
Morin, INSERM, Universitι de ROUEN, Rouen Cedex, France) The Biotrin OxyDNA Assay provides a new simple method for
detecting the impact of compounds with the potential of causing genotoxicity
early in the drug development process. The Biotrin OxyDNA Assay is a
fluorescent protein-binding assay for detecting oxidative DNA damage in situ. It can be used to detect
oxidative DNA damage by direct staining or FACS techniques. Data on the use of the Biotrin OxyDNA Assay
to detect oxidative DNA damage in cell culture caused by free radicals and
other causes of oxidative stress will be presented including endothelial cells,
epithelial kidney cells and organotypic cultures of lung tissue.
11:30 12:15 PM
Quantitative Structure-Toxicity Relation-ships by
Accelerated Cytotoxicity Mechanism Screening. (Peter J. O'Brien, University of Toronto, Toronto, Ontario,
Canada)
"Accelerated Cytotoxic Mechanism Screening" (ACMS) techniques using
freshly isolated rat hepatocytes have been compared with high throughput
toxicity screening methods using human HepG2 cells to derive quantitative
structure toxicity relationship equations for a variety of phenols, catechols,
polyphenols, aniline derivatives and NSAIDS. The role of the chemical
parameters of these agents including log P, pKa, and redox potential for
predicting their cytotoxic dose, their effects on mitochondrial function and
their susceptibility to metabolic activation to reactive intermediates will be
described. These studies should provide
a foundation for predicting the toxic potential and possible cytotoxic
mechanisms of new drug candidates. New in vitro screening methods will be
proposed.
12:15 1:30 PM
- LUNCH BREAK
1:30 1:45 PM
Screening of Drug Candidates for Their Potential to Induce Apoptosis and Necrosis (Vangala Subrahmanyam, Purdue Pharma, Ardsley, NY) Drugs induce primarily two types of cell death widely recognized as apoptosis and necrosis. Apoptosis is considered as non-harmful event where a damaged cell triggers its own demise without inducing inflammation. On the other hand necrotic cell death may release chemokines that activate inflammatory reactions and result in a cascade of events that will result in organ failure. Another mode of cell death recently recognized is apoptotic necrosis, maybe activated via defective apoptosis. This talk will focus on how cytotoxicity tests for apoptosis and necrosis may be utilized in drug discovery. The predictive value of such tests will be discussed.
1:45
2:30 PM
The Use of Bioluminescence Based Assays for Cell Viability and Apoptosis: Suitable for High Throughput Screening Applications. (S P M Crouch, T S Simmons, D A Bradbury and K J Slater. LumiTech Ltd., City Link, Nottingham, UK; Presented by Tony McCook, Biowhittaker, Gaithersburg, MD) The measurement of cellular adenylate nucleotides, using bioluminescence, has been used to develop assays for the measurement of cell proliferation/cytotoxicity, and also for the differentiation of apoptosis and necrosis. The models tested included U937 cells with camptothecin and L929 cells with TNF for cytotoxicity. There was a concentration dependent reduction in ATP bioluminescence, which correlated with loss of cell viability assessed by propidium iodide (PI) uptake (U937 p<0.0001, n=10) and crystal violet staining (L929 P<0.00001, n=8). We have investigated proliferation in a number of growth factor dependent models, mixed lymphocyte reactions and induction of human airway smooth muscle cell proliferation using a variety of agonists. In all models tested there was a significant correlation (p<0.0001) with tritiated thymidine incorporation. More than 35 different models were used for apoptosis/necrosis, in both adherent and suspension cell lines and primary cell cultures. Here the amount of cellular ADP present relative to ATP was determined (ADP:ATP ratio). Data using HL-60, Cem-7, Jurkat and U937 cell lines and apoptosis inducing agents, showed a significant correlation (p<0.001) with TUNEL and estimation of the sub Go fraction using PI staining. Ratio data also correlated with disruption of the mitochondrial transmembrane potential using the fluorescent cyanine dye JC-1 (P<0.001). Use of the broad spectrum caspase inhibitor ZVAD-FMK showed a reduction in the ADP:ATP ratio correlating with caspase 3 activity and expression of phosphatidyl serine on the cell surface, as determined by Annexin V binding and flow cytometry. To confirm the applicability of these tests for high throughput screening, Z statistics were performed with the Z consistently greater than 0.8. All assays are safe, simple and easy to perform with minimal (15 minutes) preparation time.
2:45 3:30 PM
In Vitro Stem Cell Hemotoxicity Assays as a Predictive Tool for Continuously Proliferating Cell Systems (Ivan N. Rich, HemoGenix LLC; Irmo, SC) The five continuously proliferating systems of the body are the lympho-hematopoietic system, the gastrointestinal tract, the germ cells of the reproductive organs, the dermal cells of the skin and the corneal epithelial cells of the eye. All possess stem and progenitor cell populations that are extremely sensitive to any perturbation, especially from anti-neoplastic drugs. The stem and multi-lineage cell populations of the hematopoietic system provide a readily amenable, cost-effective and highly sensitive cell-based and surrogate screening system to predict agent toxicity. New high throughput and chronotoxicity assays are being developed.
3:30 -
4:15 PM
Round Table Discussion
END OF CONFERENCE
POSTER PRESENTATIONS:
Poster Presentations are always encouraged. Please submit your poster abstract for
approval by the organizing board by
November
30th.
Submit posters to Nola Mahaney, Director of
Administration; ISE, Inc.; 7061 Deepage Drive, Suite 102; Columbia, MD 21045 or
fax at (410) 309-9014 or email file attachment to nola@isciencex.com.
Participating
Exhibitors as of June 25, 2001:
