1 research outputs found
Universal and Quantitative Method To Evaluate Inhibitor Potency for Cysteinome Proteins Using a Nonspecific Activity-Based Protein Profiling Probe
Recently, there have
been a limited number of new, validated targets
for small-molecule drug discovery in the pharmaceutical industry.
Although there are approximately 30 000 genes in the human
genome, only 2% are targeted by currently approved small-molecule
drugs. One reason that many targets remain neglected by drug discovery
programs is the absence of biochemical assays enabling evaluation
of the potency of inhibitors in a quantitative and high-throughput
manner. To overcome this issue, we developed a biochemical assay to
evaluate the potency of both reversible and irreversible inhibitors
using a nonspecific thiol-labeling fluorescent probe. The assay can
be applied to any targets with a cysteine residue in a pocket that
can accommodate small-molecule ligands. By constructing a mathematical
model, we showed that the potency of compounds can be quantitatively
evaluated by performing an activity-based protein profiling assay.
In addition, the validity of the theory was confirmed experimentally
using epidermal growth factor receptor kinase as a model target. This
approach provides an assay system for targets for which biochemical
assays cannot be developed. Our approach can potentially not only
expand the number of exploitable targets but also accelerate the lead
optimization process by providing quantitative structure–activity
relationship information