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ASSAY DEVELOPMENT IN DRUG DISCOVERY

ASSAY DEVELOPMENT IN DRUG DISCOVERYI ndira Padmalayam, DISCOVERY Division, Southern Research Institute, Birmingham, Alabama 35205 The Drug DISCOVERY Process:The ASSAY DEVELOPMENT StageAssay developmentPrimary assays Secondary assaysTarget IdentificationTarget ValidationAssayDevelopmentHTSLead IdentificationLead OptimizationDevelopment Hit validationChemistryStructure-Activity Relationship (SAR)bioavailability (PK, ADME), toxicityIn vivo efficacyPre-clinicalGLP-ToxClinicalTarge t IdentificationTarget ValidationAssayDevelopmentLead Optimization DevelopmentHTSNon-HTSA ssay DEVELOPMENT : A critical part of the hit DISCOVERY process HITS A hit is a compound which has the desired activity in acompound screen and whoseactivity is confirmed upon retestingAdapted frrom: Hughes, (2011), British Journal of Pharmacology, 162 1239-1249 Secondary assaysPrimary assaysAssay DevelopmentPre-Roadblocks to faster ASSAY DEVELOPMENT Why is this a bottleneck?

Fluorescence-based assays (contd.) FRET: Florescence Resonance Energy Transfer Half life: 1 to 10 nano secs • Principle: Two fluorophores: Donor and Acceptor-Based on transfer of energy between donor and acceptor-Distance is critical Far: No energy transfer no FRET Close: Energy transfer from donor to acceptor FRET

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