2. DuPont Pharmaceuticals Company Utilization of Spotfire™ in Quality Assessment & Analysis of Screening Data Thomas D.Y. Chung, Ph.D. Sr. Director, Leads Discovery Operations Dupont Pharmaceuticals Company Wilmington, DE © 2001, DuPont, Inc. - All Rights Reserved.

3. DuPont Pharmaceuticals Company Target Number of Screens (Attrition Model) 1 7-1015-20 5-7 40-50 20-30 NDA Pre- IND Med CHEM Early CHEM Qualified LEAD New HTS No. of Cmpds 200 - 500K Decision Cost & Quality $$$$$

4. DuPont Pharmaceuticals Company Spotfire Utilization in DuPont Pharma Leads Discovery Data QC review and artifact alerts Survey screen performance & consistency Decision support for compound selectivity assessment Allows easy selection, justification, and communication of desired “hits” Summary views on multivariate data for HT- ADME and Pharmaceuticals Properties Profiling

5. DuPont Pharmaceuticals Company Compound Progression Schema 1º HTS w/ or w/o Counterscreen (singlet or replicate) Plate & Run QC/ Post run filtering Confirmations of solutions & Dose Response Curves (DRC) Review of stats, false positives & negatives, & self-consistency of %I or %A and IC50 or EC50 Check chemical structure for obvious flaws HIT SELECTION, CONFIRMATION & DOSE-RESPONSE Obtain independent sample (check lots), powder if available. Confirm purity, identity & bioactivity of major peaks. LC/MS Refine dose-response curve SAMPLE POTENCY & IDENTITY CONFIRMATION Look for nascent SAR - similarity & substructure Selectivity & P3 profiles & liabilities Chemical attractiveness & starting point for optimization Bioassay in TA in vitro & in vivo models NEW LEAD SERIES? LEAD SAR, QUALIFICATION & PRIORITIZATION I. CONFIRMED HIT Is solution activity confirmed, potent (selective?) & chemically interesting? II. CONFIRMED ACTIVE Is independent sample (solid or liquid) potent, pure & structure correct? III. QUALIFIED LEAD SERIES Is structure “druggable”? Acceptable selectivity, P3 properties, chemical tractability, TA bioeffect

6. DuPont Pharmaceuticals Company Liquid Handling Validation

7. DuPont Pharmaceuticals Company Assay Run QC - Spotfire Pro  Stack view Good run Stack view Severe Edge Effects

8. DuPont Pharmaceuticals Company Assay Confirmation QC- Spotfire™.net

9. DuPont Pharmaceuticals Company Run QC - Plate Drill-down Spotfire™.net

10. DuPont Pharmaceuticals Company Selectivity Profiling

11. DuPont Pharmaceuticals Company Compound Number Analytical Quality Low Medium Very High HTS Specificity/Chem Analysis Solub/SPB/ Bioavail /Metab HTS to SAR SAR 100000-1000000 50-750 25-400 2-5 20-200 1-5 Solub/SPB/Bioavail/Metab Primary Screen Lead Qualification Progression v

12. DuPont Pharmaceuticals Company Desireable Drug-Like Characteristics Pharmacologically active and specific Favorable oral bioavailability soluble membrane permeable reasonable uptake not substrate of efflux-pumps Desirable pharmacokinetic properties distribution and clearance 60%-90% serum protein binding metabolism low toxicity no drug-drug interactions not substrates for cytochromes P450

13. DuPont Pharmaceuticals Company HT- Pharmaceutical Properties Profiling Purity & Bioactivity Correlation CYP 3A4, 2D6 & 2C9 Inhibition Pgp Efflux Inhibition Serum Binding Solubility* - nephelometry or UV/LC Metabolism* - liver microsomes/cells Permeability - surface activity (tension)

14. DuPont Pharmaceuticals Company Acknowledgments Ilona Kariv Louis Coudurier Gerard McGeehan