Quantitative high-throughput screening: a titration-based approach that efficiently identifies biological activities in large chemical libraries.

By James Inglese, D. Auld, A. Jadhav, Ronald L. Johnson, Anton Simeonov, Adam Yasgar, Wei Zheng, C. Austin. 2006

773 citations recorded by the source corpus.

High-throughput screening (HTS) of chemical compounds to identify modulators of molecular targets is a mainstay of pharmaceutical development. Increasingly, HTS is being used to identify chemical probes of gene, pathway, and cell functions, with the ultimate goal of comprehensively delineating relationships between chemical structures and biological activities. Achieving this goal will require methodologies that efficiently generate pharmacological data from the primary screen and reliably profile the range of biological activities associated with large chemical libraries. Traditional HTS, which tests compounds at a single concentration, is not suited to this task, because HTS is burdened by frequent false positives and false negatives and requires extensive follow-up testing. We have developed a paradigm, quantitative HTS (qHTS), tested with the enzyme pyruvate kinase, to generate concentration-response curves for >60,000 compounds in a single experiment. We show that this method is precise, refractory to variations in sample preparation, and identifies compounds with a wide range of activities. Concentration-response curves were classified to rapidly identify pyruvate kinase activators and inhibitors with a variety of potencies and efficacies and elucidate structure-activity relationships directly from the primary screen. Comparison of qHTS with traditional single-concentration HTS revealed a high prevalence of false negatives in the single-point screen. This study demonstrates the feasibility of qHTS for accurately profiling every compound in large chemical libraries (>10(5) compounds). qHTS produces rich data sets that can be immediately mined for reliable biological activities, thereby providing a platform for chemical genomics and accelerating the identification of leads for drug discovery.

Subfields: Biomedical Engineering, Computational Theory and Mathematics, Molecular Biology, Computational Chemistry & Drug Discovery

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Citations in the corpus

Molecular Docking and Structure-Based Drug Design Strategies (2015, background): “These programs are useful in predicting important properties related to biological activity, such as solubility, protonation state and molecular volume [132].”
Toxicity Prediction using Deep Learning (2015, background): “These experiments are repeated with varying concentrations of the chemical compound, which allows to determine dose-response curves (Inglese et al., 2006).”
Improving the Human Hazard Characterization of Chemicals: A Tox21 Update (2013, methodology): “…at the NCGC “proof of principle” libraries of 1,408 and 1,462 compounds, respectively, with each compound dissolved and stored in dimethyl sulfoxide (DMSO), to be evaluated for activity in 1,536well plate quantitative high throughput screens (qHTS), as described by Inglese et al. (2006).”
High-throughput screening assays for the identification of chemical probes. (2007, background): “3a , quadrants i and ii) and may be rescued for HTS using replicate or concentration-dependent format”
TOXICITY TESTING IN THE 21ST CENTURY: A VISION AND A STRATEGY (2010, methodology): “Quantitative high-throughput assays can be used to expand the coverage of the universe of new and existing chemicals that need to be evaluated for human health risk assessment (Roberts, 2001; Inglese, 2002; Inglese et al., 2006; Haney et al., 2006).”
A Homogeneous, High-Throughput Assay for Phosphatidylinositol 5-Phosphate 4-Kinase with a Novel, Rapid Substrate Preparation (2013, methodology): “Pilot Screen Validation of the PI5P4Ka assay was performed by a qHTS (quantitative HTS, [25]) against the library of pharmacologically active compounds (LOPAC(1280), Sigma-Aldrich) arrayed as a fivepoint titration series (152, 76, 15, 3, and 0.”
Apparent activity in high-throughput screening: origins of compound-dependent assay interference. (2010, background): “Originally developed as a drug discovery methodology by pharmaceutical companies for identification of chemical entities with potential therapeutic value, HTS has branched into the academic sector as a technique to discover chemical probes, which are applied as research tools to study a biological t”
The NCGC Pharmaceutical Collection: A comprehensive resource of clinically approved drugs enabling repurposing and chemical genomics (2011, methodology): “2% [hits classi ed according to (66)], above the average rate of 1.”