Wednesday, October 13, 2004

 

8:00 AM – 9:00 AM –REGISTRATION

 

8:00 AM – EXHIBITS

 

 

8:50 AM – 9:00 AM – Exhibitor Presentation-BD Biosciences

 

9:00 AM – 9:30 AM

PAMPA "in combo" Strategies to Predict Caco-2 and the Blood-Brain Barrier Permeability (Alex Avdeef, pION INC; Woburn, MA) PAMPA (parallel artificial membrane permeability assay) comparative studies will be discussed in detail, featuring --

·          Double-Sink^TM technology (pH gradient, lipophilic gradient, 20% phospholipid mixture in alkane solvent, with some negatively-charged lipid components)

·          individual-well stirring to lower assay time to 10 minutes for lipophilic, sparingly-soluble molecules

·          cosolvent method to allow study of molecules with solubility as low as 6 ng/mL (amiodarone), while still using UV detection

·          in vivo - in vitro correlation between PAMPA and rat in situ perfusion data allows for MAD (maximum available dose) analysis earlier in Discovery

 

9:30 AM – 10:00 AM

Application of Artificial Membranes (e.g. PAMPA) in the Evaluation of Drug Absorption (Hanlan Liu, Genzyme Drug Discovery and Development; Cambridge, MA) Parallel artificial membrane permeability assay (PAMPA) utilizes a simple phospholipid-coated filter disc to measure membrane permeability.  It was developed as an alternative to the low throughput Caco-2 cell permeability assay used to correlate passive permeability with in vivo oral absorption.  This talk will focus on addressing issues found when implementing the assay using real discovery-level compounds.  A few examples of PAMPA assay for evaluating in vivo drug oral absorption/oral bioavailability in both pre-clinical species and human will also be presented.

 

10:00 AM – 10:30 AM

Exploratory Biopharmaceutical Classification System (BCS) Approaches:  The Use of In Silico and High Throughput Solubility and Permeability Methodologies (Marcus Brewster, Johnson & Johnson; Beerse, BELGIUM) Early screening of the drug pipeline is essential in improving the quality of the drug candidate as well as decreasing attrition.  One useful method in this regard is the estimation of a BCS value at early stages based on in silico and high throughput solubility and permeability assessments.  Permeability can be evaluated using artificial membrane techniques including PAMPA Comparisons of PAMPA and CACO-2 permeability have been completed demonstrating the general utility of these systems.

 

10:30 AM – 10:45 AM – BREAK

 

10:45 AM – 10:55 AM – Exhibitor Presentation – LiConic Instruments

 

10:55 AM – 11:25 AM

Application of Intact Cell Systems (e.g. Caco-2) in the Evaluation of Drug Absorption (Dooman Oh, Amgen Inc.; Thousand Oaks, CA) Gastrointestinal absorption of a drug candidate is a complex process, influenced by physicochemical properties, inherent permeability, modulation by uptake and/or efflux transporters, and its formulation. Hence in vitro models of absorption (e.g. Caco-2 monolayer) have become essential components of lead optimization process. Specialized cell systems expressing transporters are also being used widely for answering specific questions related to absorption, distribution, and elimination.

 

11:25 AM – 11:55 AM

Caco-2 Cell Model: A Tool to Evaluate the Absorption Mechanism of Drug Candidates (Cindy Xia, Millennium Pharmaceuticals, Inc.; Cambridge, MA) The Caco-2 cell line was first introduced to the pharmaceutical industry in 1989.  It has since been used extensively as an in vitro model of human intestinal epithelium for studies of intestinal drug absorption/transport and metabolism.  Many active transport systems such as P-glycoprotein (Pgp), multi-drug resistance protein 2 (MRP2), peptide transporter (PEPT1), and nucleoside transporter (NT) normally found in small intestinal enterocytes have been characterized in Caco-2 cells.  In this presentation, the presence of breast cancer resistant protein (BCRP) in Caco-2 cells and the usage of Caco-2 cell model in preclinical discovery and development will be assessed.

 

11:55 AM – 12:25 PM

From ABC to SLC:  applications of drug transporters in the analysis of drug absorption and distribution (Charles Crespi, BD Biosciences; Newport, PA) Multiple human (and animal) SLC and ABC transporters have been heterologously expressed in xenopus oocytes, LLC-PK1 cells and baculovirus/insect cells.  These materials enable important applications in drug discovery and distributions.  For example, the potential for drug-drug interactions at the level of drug transport can be analyzed, species difference in affinity or rates of transport can be analyzed and drug transport can be subjected to reaction phenotyping-type analysis.  The systems will be described and examples provided.

 

 

12:55 PM – 2:30 PM – LUNCH BREAK

 

 

2:30 PM – 2:40 PM – Exhibitor Presentation – Qualyst

 

2:40 PM – 3:10 PM

Application of Cryopreserved Human Hepatocytes in Drug Discovery (David B. Duignan, Pfizer Global Research & Development; Groton, CT) Cryopreserved human hepatocytes represent a useful in vitro model to assess hepatic disposition in drug discovery.  This talk will highlight current and potential applications of cryopreserved human hepatocytes in drug metabolism studies, including the assessment of metabolic liability and biliary excretion.

 

3:10 PM – 3:40 PM

In Vitro Methods and In Silico Approaches for Drug Metabolism Support of Drug Discovery (Tim Carlson, Amgen; Thousand Oaks, CA) In vitro assays for drug metabolism and permeability have been used extensively in the pharmaceutical industry to support drug discovery programs.  The data from these assays can be used to prioritize compounds for further evaluation in vivo.  We have focused on optimizing the throughput and the robustness of these assays to increase the number and the diversity of compounds that can be profiled.  An additional goal is to use the in vitro data to build and implement predictive computational models for use in the design of compounds with more optimal properties.  Efforts towards optimizing liver microsomal metabolic stability and cytochrome P450 inhibition potency will be discussed.

 

3:40 PM – 4:10 PM

Intrinsic Clearance Determinations: Microsome, S9, or Hepatocyte (Chuang Lu; Millennium Pharmaceuticals, Inc.; Cambridge, MA) Determining intrinsic clearances (Cl_int) of drug molecules in the early discovery stage would help project teams to confirm, first of all, if metabolism is the main clearance pathway when comparing the Cl_int with the total body clearance Cl in vivo.  It also helps in rank ordering metabolic stabilities of drug molecules.  Lastly, it helps in assessing species differences in the metabolic clearance and projecting the clearance of drug candidates in humans.  The advantages and limitations of these three commonly used in vitro systems for determining intrinsic clearances will be discussed.  Comparison of freshly isolated vs. cryopreserved hepatocytes and the use of hepatic and extra-hepatic fractions in the clearance determination will also be presented.

 

4:10 PM – 4:25 PM – BREAK

 

4:25 PM – 4:35 PM – Exhibitor Presentation - CellzDirect

 

4:35 PM – 5:05 PM

Current Strategies for the Use of Primary Hepatocytes in Drug Development: Advantages and Challenges for their Application in Drug Metabolism and Drug-drug Interactions (Geraldine Hamilton, CellzDirect; Pittsboro, NC) Species differences in drug metabolism, enzyme regulation and response to toxic insult preclude easy extrapolation from animal model systems to humans.  Accordingly, human-relevant in vitro model systems are an essential part of the drug development process, and primary human hepatocytes have become the ‘gold standard’ for many DMPK studies.  The current lecture will focus on strategies for using primary hepatocytes to determine metabolic stability and enzyme induction. A critical evaluation of new technologies and future challenges will also be presented.

 

5:05 PM – 5:35 PM

Induction of P450 and Other DMEs:  Role of Nuclear Receptors and Impact on Drug Metabolism and Drug Development (David J. Waxman, Boston University; Boston, MA) Many foreign chemicals, including pharmaceuticals, induce the expression of cytochrome P450 and other drug metabolizing enzymes in liver and other tissues.  This presentation will discuss 1) the role of nuclear receptors in mediating P450 induction, 2) in vitro methods used to screen for induction, and 3) the impact of P450 induction on drug metabolism, drug interactions and drug development.

 

5:35 PM – 6:05 PM - ***NEW SPEAKER***

Tackling the “E” in ADMET: An In Vitro Sandwich-Cultured Hepatocyte System for Biliary Excretion and Hepatic Uptake (Kenneth Brouwer, Qualyst, Inc.; Research Triangle Park, NC) Rapid hepatic uptake and extensive biliary clearance limit the therapeutic potential of many drug candidates.  Active transport systems are responsible for the movement of many compounds across the sinusoidal membrane into the hepatocyte.  The parent compound and/or metabolites may undergo secretion into bile by primary, active ATP-dependent export pumps located in the canalicular membrane.   While several models (isolated perfused livers, suspended hepatocytes) have been used to investigate the hepatic disposition of compounds, a reliable moderate-throughput in vitro method to predict the hepatobiliary disposition (hepatic uptake and biliary excretion) of drug candidates and metabolites has been lacking.  Qualyst has developed B-CLEAR^™, an in vitro sandwich-cultured hepatocyte system for biliary excretion and hepatic uptake.  The B-CLEAR^™ system maintains hepatocytes in culture between two layers of gelled collagen (a sandwich configuration), resulting in the formation of extensive, functional canalicular networks.  Recent studies have indicated that the in vivo biliary clearance in rats of numerous model substrates can be predicted accurately by the in vitro B-CLEAR^™ system.  An important step in defining potential mechanisms of aberrant pharmacokinetic disposition of drug candidates is to characterize the hepatobiliary disposition of compounds including the extent of hepatic uptake and excretion, and identifying the transporters involved in hepatobiliary disposition.  A major advantage of B-CLEAR^™ is the ability to investigate alterations in hepatic transport systems in human hepatocytes, evaluate drug-drug interaction potential, and predict the hepatobiliary disposition of drug candidates at an earlier stage in the drug discovery process than possible using traditional animal studies.

 

Session 2 will be continued the next day.

END OF DAY

Thursday, October 14, 2004

 

 

8:00 AM – 9:00 AM –REGISTRATION

 

8:00 AM – EXHIBITS

 

8:50 AM – 9:00 AM – Exhibitor Presentation – XenoTech LLC

 

9:00 AM – 9:30 AM

Stable Cell Lines For Determining Induction and Inhibition of Biotransformation Systems (Judy Raucy, 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.  Such alterations can result in adverse reactions stemming from drug-drug interactions.  A simplified and reliable tool for detecting the ability of candidate drugs to alter CYP3A4 levels or inhibit its catalytic activity was developed by stably integrating into HepG2 cells, the human pregnane X receptor (hPXR) and a luciferase vector harboring the CYP3A4 enhancers.  Treatment of these stable transformants (DPX-2) with various concentrations of inducers including rifampicin, mifepristone, troglitazone, methoxychlor, and kava produced dose-dependent increases in luciferase expression that correlated to CYP3A4 mRNA levels.  To determine the utility of DPX2 cells for identifying inhibitors of CYP3A4 catalytic activity, luciferin BE activity was measured in the presence of various concentrations of ketoconazole or erythromycin.  Both agents exhibited dose dependent decreases in CYP3A4 enzyme activity.  Collectively, DPX2 cells can be used to identify xenobiotics that induce or inhibit CYP3A4 enzyme activity in a high throughput manner demonstrating the applicability of this cell line to early stage drug development. 

 

9:30 AM – 10:00 AM

Latest assessment of Fa2N-4 immortalized human hepatocytes for use in drug discovery and development (Andrew Parkinson, Xenotech, Lenexa, KS) Efforts are underway to evaluate the potential of immortalized human hepatocytes for optimal use in drug discovery and development applications.  This novel test system offers an unlimited supply of cells, predictable scheduling and consistent results over time.  The latest characterization data comparing the Fa2N-4 immortalized hepatocytes to primary cultures of human hepatocytes will be presented.

 

10:00 AM – 10:30 AM

In Vitro - In Vivo Scaling of Pharmacokinetic Drug Interactions: As Good as it Gets? (David J. Greenblatt, Lisa L. von Moltke, Tufts-New England Medical Center) During the last 15 years, an extensive research investment has been made into development of in vitro models to anticipate and predict clinical pharmacokinetic drug interactions.  Such models, if adequately validated, could potentially facilitate improved targeting of time and resources for clinical studies, and even allow some studies to be avoided altogether.  The core assumption of scaling approaches is that an in vitro inhibitory K_i or IC₅₀ versus a specific substrate can be combined with the in vivo exposure to inhibitor (such as inhibitor concentration [I] or inhibitor AUC) to yield a quantitative prediction of the relative clearance decrement (compared to control) of the same substrate when coadministered with the inhibitor.  A survey of scaling studies over the past decade reveals many highly accurate predictions, as well as numerous dismally inaccurate preductions.  Identified grounds for poor prediction include in vitro methologic factors (such as nonspecific binding of substrate or inhibitor, mechanism-based inhibition, solvent effects, substrate-specific inhibition) or physiologic factors (plasma or tissue binding, concentrative hepatic uptake, route-dependent clearance, presystemic extraction, enteric metabolism, concurrent efflux transport).  Modification of models to account for such factors may improve predictive accuracy, but generally modifications are applied in retrospect rather than prospectively.  This presentation reviews the current status and future prospects for in vitro-in vivo scaling of drug interactions.

 

10:30 AM – 10:45 – BREAK

 

10:45 AM – 10:55 AM – Exhibitor Presentation – Tissue Transformation Technologies

 

 

 

11:25 AM – 11:55 AM

In vitro toxicity assays:  From human hepatocytes to IdMOC (Albert P. Li, APSciences Inc., Baltimore, MD) Human hepatocytes represent a relevant model for the evaluation of human hepatotoxicity, as they possess human specific metabolism and are the in vivo targets of most hepatotoxic drugs.  The application of human hepatocytes in toxicity evaluation will be discussed, including the measurement of cytotoxicity and the application of toxicogenomics.  The successful application of human hepatocytes leads to the development of a “complete” in vitro toxicity assay using a novel culture system:  the Integrated Discrete Multiple Organ Culture (IdMOC) with primary human cells from multiple organs (e.g. hepatocytes, renal proximal tubule cells, aortic endothelial cells, astrocytes, small airway epithelial cells) for the evaluation of multiple organ toxicity.

 

11:55 AM – 12:25 PM

Preliminary Screening of Compound Libraries to Distinguish Between Potential Genotoxic and Non- genotoxic Carcinogens in Rat Livers Cells Using Gene Expression Profiling (Gordon Vansant, Althea Technologies; San Diego, CA) When distinct gene expression responses are reproducible for a series of related compounds the patterns are defined as “gene signatures” and are useful for compound screening.  There are up to 100 gene responses associated with the impact of genotoxic compounds and we are using these genes to develop a “genotoxic gene signature” in vitro.  Analysis of the gene expression patterns produced by 100’s of different compounds has resulted in the identification of expression profiles that are characteristic of genotoxic as well as non genotoxic compounds. 

 

12:25 PM – 2:00 PM – LUNCH BREAK

 

2:00 PM – 2:10 PM – Exhibitor Presentation – APSciences, Inc.

 

2:10 PM – 2:40 PM

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 (Carl Alden, Millennium Pharmaceuticals, Inc.; Cambridge, MA) Opportunities for improvement in efficiency from a toxicology and safety pharmacology perspective in pharma R&D lie in leveraging “best in class” technology and early integration with medicinal chemistry and pharmacology activities during H2L and early lead optimization. Liver toxicity represented the predominant reason for drug withdrawals and black boxes from 1975 to the end of the century but is now supplanted by functional side effects (involving CNS and cardiovascular systems). H2L and early LO toxicity and safety pharmacology screening, to be practical on a routine basis, must be reasonably high through put (one person conducting 20 to 50 analyses per week) and low in test agent consumption (less than 10 mg per assay). To meet this challenge we have adopted a “discovery assay by stage” (DABS) paradigm that clearly delineates the timing and type of assay required for selection of the lead series and for representatives from the lead series to progress in the discovery continuum. 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.

 

2:40 PM – 3:10 PM

Tissue 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.

 

3:10 PM – 3:25 PM – BREAK

 

3:25 PM – 3:55 PM

Bioluminescent Methods for ADME/Tox (Michael Curtin, Promega Corporation, Madison, WI) We have harnessed the advantages of bioluminescence to configure sensitive, simple to perform, homogeneous, high through-put assays for ADME/Tox applications. Systems have been developed for measuring the cytotoxicity of test compounds, their capacity to induce apoptosis, their impact on various CYP450 activities and on multi-drug transporters. In each case an add-mix-measure approach is used as the basis for robust and highly reproducible assays.

 

 

END OF CONFERENCE

 

 

POSTER PRESENTATIONS:

 

Poster Presentations are always encouraged.  Please submit your poster abstract for approval by the organizing board by August 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 Tuesday October 12 or before the first session begins on Wednesday, October 13^th. 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, 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

Sheraton Commander Hotel, Cambridge; 16 Garden Street, Cambridge, MA, USA. The original hotel of Harvard Square, opened in 1927.  The Commander is situated on historic Cambridge common near the site where General George Washington took command of the continental Army in 1775. With Cambridge being the center of history and culture, the Sheraton Commander Hotel is the perfect vantage point from which to experience the sites and sound of this wonderful city.  We are minutes away from prestigious academic institution such as Harvard University, Radcliffe, Lesley University, Tufts University and M.I.T.

 

 

Hotel Information

Please contact the hotel directly to make your accommodation reservations if necessary.  $220.00 per single king-size or 2 double bedroom.  There is an additional charge for extra persons.  Please make your hotel reservation directly by telephone at 617.547.4800; Fax: 617.234.1396. or 888.627.7121

On-line Reservations:  http://www.sheratoncommander.com/cambridge.htm. 

 

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 $250.00). 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 September 13 2004. Please submit cancellation and refund requests including transferring of registration to: Fax:  410-869-9560; E-mail: nola@isciencex.com; Cancellation Deadline: September 13, 2004

 

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.

 

Payment Methods Accepted:  Check, American Express, VISA, Master Card.

 

IVAT-2004 Conference: US $1500.00 ______________

 

SPECIAL DISCOUNTS:

Both **Training Course-Boston & IVAT-2004: US$2500.00

 

One Training Course & IVAT-2004: US $2000.00

(see Training Course Agenda for further discounts)

 

**ISE Training Course in Drug Development; October 11-12, 2004, Boston, MA, USA

 

Exhibitors: US $2000.00

 

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

 

Academic/Government participants will receive a 50% discount.