Universal and Quantitative Method To Evaluate Inhibitor Potency for Cysteinome Proteins Using a Nonspecific
Tomoya Sameshima1, Yukiya Tanaka1, Ikuo Miyahisa1
1Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited , 26-1, Muraoka-higashi 2 chome, Fujisawa, Kanagawa, Japan.
Biochemistry
|May 19, 2017
Summary
A new biochemical assay uses a fluorescent probe to quantify inhibitor potency for drug discovery targets, including those previously lacking assays. This method accelerates the identification and optimization of new small-molecule drugs by enabling high-throughput screening.
Area of Science:
- Biochemistry
- Drug Discovery
- Chemical Biology
Background:
- Limited validated targets for small-molecule drug discovery, with only 2% of human genes currently targeted by approved drugs.
- Absence of quantitative, high-throughput biochemical assays hinders the evaluation of inhibitors for many potential drug targets.
- Need for novel assay development to expand the druggable genome and accelerate pharmaceutical research.
Purpose of the Study:
- To develop a novel biochemical assay for quantitative evaluation of inhibitor potency against drug targets.
- To enable high-throughput screening for both reversible and irreversible inhibitors.
- To provide a method for targets lacking existing biochemical assay systems.
Main Methods:
- Development of a biochemical assay utilizing a nonspecific thiol-labeling fluorescent probe.
- Application of the assay to targets containing a cysteine residue accessible to small-molecule ligands.
- Mathematical modeling to quantitatively evaluate inhibitor potency using activity-based protein profiling.
- Experimental validation using epidermal growth factor receptor kinase as a model target.
Main Results:
- A novel assay system was successfully developed and validated for quantifying inhibitor potency.
- The assay demonstrated applicability to targets with accessible cysteine residues.
- Quantitative structure-activity relationship (QSAR) information can be derived, aiding lead optimization.
Conclusions:
- The developed assay system significantly expands the range of druggable targets by enabling potency evaluation.
- This approach accelerates the drug discovery pipeline by facilitating high-throughput screening and lead optimization.
- The method provides a crucial tool for targets previously inaccessible to traditional biochemical assays.
