1	Stability of Compounds Stored in TekCel Plate Management System: Application of New Approaches for Ultra-High-Throughput Analysis of Compound Stability. James A. Connelly Ph.D. Director of Library Production and QC Aventis Combinatorial Technologies Center Tucson, Arizona
2	Overview Integration of ACTC Combinatorial Chemistry Library Collection in Aventis Lead Generation Operations Characteristics of ACTC and Aventis Collections Compound Logistics Support of Multiple Screening Strategies Integration into Global Compound Logistics Network (HPU) TekCel System at ACTC and Future Plans Comparison of Compound Purity in DMSO Solutions Stored in TekCel PlateStore versus Freezers New Approaches for Surveillance of Purity Quality and Stability of Combinatorial Libraries Ultra High-throughput Chromatography (Nanostream Veloce) Flow-Injection Mass Spectrometry (Phoenix S&T) Combined uHTC and FI-MS Requirement for Highly Standardized Methods
3	Aventis Collection Attributes Many “Solids” of Relatively Large Quantity Liquid Screening and CP Stocks Created from Solids Stocks Historical MedChem Compounds from HEP and COP Programs Purchased Compounds Quality Criteria >80% Purity Primary Stocks Handled as Single Containers Stored in a Large Enterprise Level System Produced as Liquid Stocks Scaffold Based Combinatorial Libraries Limited Compound Quantity Variable Purity 75%>50% Purity Plate Based Samples Relatively Large libraries Storage in Racks in Freezers Manual Access for Screening and Cherry-picking
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5	Aventis Combinatorial Library Collection Naturally Exists as a Modular Set of Libraries Produced in 96 Well Format DMSO Liquid Stocks 384 Well Plates as Screening and Cherry Picking Stocks
6	Aventis Screening Strategies
7	Global Compound Logistics Network
8	ACTC Compounds Delivered into Many Processes Assembly of Aventis Rational Screening Sets Compound Testing after Virtual Screening Profiling, Biological, Physical and Pharmacological ACTC (Local) Screening Cherry-Picks for Activity and Structure Confirmation Local and Global Aventis Screening at Major Sites Single Library Requests Multiple Library Requests Selected Compound Subsets Screening Follow-up Activities
9	Solutions for Integration of ACTC Collection Implement Global Compound Logistics Software and Database Structuring ACTC Collection into 3 Stocks Screening Stock to Enable 384/1536 Pin-Tool HTS Cherry-Picking and Library Copy Stock High Throughput, Rapid Response, Low Volume Needs Archive Stock Low Throughput, Refilling and Larger Volume Needs TekCel PlateStore Systems for HTS and CP Activities Future: Consideration of TubeStore/TekBench
10	TekCel Solution Initially Implemented at ACTC
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12	Output of TekCel PlateStore System
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14	Compound Purity/Stability Comparison One CC Library Analyzed, TL118 (12,232 Compounds) 10 mM DMSO Solutions Stored for 1.5 Years Under Two Different Conditions TekCel PlateStore Storage, Nitrogen Purge, Minus 10 degrees C Accessed Routinely for Cherry-Picking Standard Storage Freezers in Lidded Plates Accessed 16 Times for Screening Comparative Analysis of 384 Samples at Endpoint
15	Compound Stability/Purity (TekCel vs Standard)
16	Purity Variance (TekCel vs Standard)
17	Approaches for Compound QC/Stability Standard LC/MS (48 Samples/Day, MUX 384 Samples/Day) Information Rich, Costly, Slow, Not Generally Standardized Throughput Not Sufficient for Large Libraries (10,000) Ultra High-Throughput Chromatography (1200 Samples/Day) Less Information, Lower Cost, More Easily Standardized Comparative Analysis Over Time Possible (If Standardizable) Flow Injection MS Analysis (Current 2000 Sample/Day) Less Information, Very Low Cost, ultra-HT, Standardizable? Comparative Analysis Over Time Possible (If Standardizable) Combined uHTC and FI-MS Need Highly Standardized Methods
18	Nanostream Microfluidic Chromatography
19	Nanostream Veloce Microfluidic Chromatography
20	Comparison of Brio vs Standard HPLC
21	Comparison of LC/MS to uHTC plus FI-MS 384 Samples from TL118 Were Analyzed Both by LC/MS and by uHTC plus FI-MS Can uHTC plus FI-MS Results Provide Equivalent results as LC/MS? Correctly Detected All Samples Without Expected Mass Purity Class Binning Illustrates Need for Method Standardization in Both Approaches uHTC plus FI-MS Could Potentially Be Used Instead of LC/MS for General Compound QC Operations Key Advantages Are Standardizability and Low Cost
22	uHTC plus FI-MS vs LC/MS (ExMass Not Found)
23	Binning into Purity Categories (uHTC vs LC/MS)
24	Binning into Purity Categories (uHTC vs LC/MS)
25	Initial QC vs Re-Analysis (Same LC/MS Method)
26	Conclusions TekCel PlateStore System Provides An Effective Mechanism for Integration of ACTC Collection into Global Aventis LG Operations TekCel PlateStore Provides Greater Stability of DMSO Solutions as Compared to Our Previous Storage Conditions New HT Analytical Technologies Hold Promise for Better Compound QC Analytical Method Standardization is Essential for Effective Analysis of Compound Purity and Stability Over Time
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