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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.
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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 $$$$$
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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
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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
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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
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DuPont Pharmaceuticals Company
Assay Run QC - Spotfire Pro
Stack viewGood run
Stack viewSevere Edge Effects
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DuPont Pharmaceuticals Company
Assay Confirmation QC-Spotfire™.net
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DuPont Pharmaceuticals Company
Run QC - Plate Drill-downSpotfire™.net
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DuPont Pharmaceuticals Company
Selectivity Profiling
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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
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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
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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)
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DuPont Pharmaceuticals Company
Acknowledgments
Ilona KarivLouis Coudurier
Gerard McGeehan