DuPont Pharmaceuticals Company Utilization of Spotfire™ in Quality Assessment & Analysis of Screening Data Thomas D.Y. Chung, Ph.D. Sr. Director, Leads.

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 CompanyWilmington, DE

© 2001, DuPont, Inc. - All Rights Reserved.

DuPont Pharmaceuticals Company

Target Number of Screens(Attrition Model)

17-1015-205-740-5020-30

NDAPre-IND

MedCHEM

EarlyCHEM

QualifiedLEAD

NewHTS

No. ofCmpds 200 - 500K

DecisionCost & Quality $$$$$

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

DuPont Pharmaceuticals Company

Compound Progression Schema1º HTS w/ or w/o Counterscreen (singlet or replicate)

Plate & Run QC/ Post run filteringConfirmations 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 & substructureSelectivity & P3 profiles & liabilities

Chemical attractiveness & starting point for optimizationBioassay in TA in vitro & in vivo models

NEW LEAD SERIES?

LEAD SAR, QUALIFICATION & PRIORITIZATION

I. CONFIRMED HITIs solution activity confirmed, potent

(selective?) & chemically interesting?

II. CONFIRMED ACTIVEIs independent sample

(solid or liquid)potent, pure & structure

correct?

III. QUALIFIED LEAD SERIESIs structure “druggable”?Acceptable selectivity,

P3 properties, chemical tractability, TA bioeffect

DuPont Pharmaceuticals Company

Liquid Handling Validation

1 3 5 7 9 11 13 15 17 19 21 23

A

D

G

J

M

P

384 Pipetting Head #1

1.0-3.0

-1.0-1.0

-3.0--1.0

-5.0--3.0

1 3 5 7 9 11 13 15 17 19 21 23

A

D

G

J

M

P

384 Pipetting Head #3

1.0-3.0

-1.0-1.0

-3.0--1.0

-5.0--3.0

CCS PLATE TRACK D 384 HEAD #1mean std. Dev. % CV

Plate1 4,907,695 258,909 5.28Plate 2 5,004,546 214,325 4.28Plate 3 5,142,413 234,261 4.56 OK!!Plate 4 5,079,111 215,448 4.24Plate 5 5,093,590 234,734 4.61

avg %CV 4.60

CCS PLATE TRACK D 384 HEAD #3mean std. Dev. % CV

Plate 6 4,497,948 634,656 14.11Plate 7 4,694,281 611,363 13.02Plate 8 4,803,828 506,096 10.54 POOR!!!Plate 9 4,352,574 678,146 15.58Plate 10 4,303,385 622,733 14.47

avg %CV 13.50

CCS PLATE TRACK D 384 HEAD #1mean std. Dev. % CV 3sd error

Plate1 2,244,378 86,409 3.90 259,227Plate 2 2,345,796 87,755 3.70 263,265Plate 3 2,319,290 83,476 3.60 250,428 VERY GOOD!!Plate 4 2,272,469 89,181 3.90 267,543Plate 5 2,289,773 87,493 3.80 262,479

avg %CV 3.80

CCS PLATE TRACK D 384 HEAD #3mean std. Dev. % CV 3sd error

Plate 6 2,311,048 89,156 3.90 267,468Plate 7 2,288,710 90,607 4.00 271,821Plate 8 2,282,292 83,585 3.70 250,755Plate 9 2,320,800 82,671 3.56 248,013 MUCH IMPROVED!!!Plate 10 2,236,199 80,283 3.60 240,849

avg %CV 3.75

DuPont Pharmaceuticals Company

Assay Run QC - Spotfire Pro

Stack viewGood run

Stack viewSevere Edge Effects

DuPont Pharmaceuticals Company

Assay Confirmation QC-Spotfire™.net

DuPont Pharmaceuticals Company

Run QC - Plate Drill-downSpotfire™.net

DuPont Pharmaceuticals Company

Selectivity Profiling

DuPont Pharmaceuticals Company

Compound Number

Analytical Quality

Low

Medium

Very High

HTS

Specificity/Chem Analysis

Solub/SPB/Bioavail/MetabHTS to SAR

SAR

100000-1000000

50-750

25-400

2-5

20-200

20-200

1-5

Solub/SPB/Bioavail/Metab

Solub/SPB/Bioavail/Metab

Primary Screen

Lead Qualification Progression

v

DuPont Pharmaceuticals Company

Desireable Drug-Like Characteristics Pharmacologically active and specific

Favorable oral bioavailabilitysolublemembrane permeablereasonable uptakenot substrate of efflux-pumps

Desirable pharmacokinetic propertiesdistribution and clearance

60%-90% serum protein bindingmetabolism

low toxicityno drug-drug interactionsnot substrates for cytochromes P450

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)

DuPont Pharmaceuticals Company

Acknowledgments

Ilona KarivLouis Coudurier

Gerard McGeehan