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Theoretical and experimental relationships between percent inhibition and IC50 data observed in high-throughput screening. | LitMetric

AI Article Synopsis

Article Abstract

The four-parameter logistic Hill equation models the theoretical relationship between inhibitor concentration and response and is used to derive IC(50) values as a measure of compound potency. This relationship is the basis for screening strategies that first measure percent inhibition at a single, uniform concentration and then determine IC(50) values for compounds above a threshold. In screening practice, however, a "good" correlation between percent inhibition values and IC(50) values is not always observed, and in the literature, there seems confusion about what correlation even to expect. We examined the relationship between percent inhibition data and IC(50) data in HDAC4 and ENPP2 high-throughput screening (HTS) data sets and compared our findings with a series of numerical simulations that allowed the investigation of the influence of parameters representing different types of uncertainties: variability in the screening concentration (related to solution library and compound characteristics, liquid handling), variations in Hill model parameters (related to interaction of compounds with target, type of assay), and influences of assay data quality parameters (related to assay and experimental design, liquid handling). In the different sensitivity analyses, we found that the typical variations of the actual compound concentrations in existing screening libraries generate the largest contributions to imperfect correlations. Excess variability in the ENPP2 assay above the values of the simulation model can be explained by compound aggregation artifacts.

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http://dx.doi.org/10.1177/1087057112455219DOI Listing

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