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Updated: Aug 12, 2025
Identification of Kinase-substrate Pairs Using High Throughput Screening
Published on: August 29, 2015
Identification of a Class of WNK Isoform-Specific Inhibitors Through High-Throughput Screening
Julita Chlebowicz1, Radha Akella1, John M Humphreys1
1Department of Biophysics, The University of Texas Southwestern Medical Center, Dallas, TX, USA.
Introduction:
WNK [with no lysine (K)] kinases are serine/threonine kinases associated with familial hyperkalemic hypertension (FHHt). WNKs are therapeutic targets for blood pressure regulation, stroke and several cancers including triple negative breast cancer and glioblastoma. Here, we searched for and characterized novel WNK kinase inhibitors.
Methods:
We used a ~210,000-compound library in a high-throughput screen, re-acquisition and assay, commercial specificity screens and crystallography to identify WNK-isoform-selective inhibitors.
Results:
We identified five classes of compounds that inhibit the kinase activity of WNK1: quinoline compounds, halo-sulfones, cyclopropane-containing thiazoles, piperazine-containing compounds, and nitrophenol-derived compounds. The compounds are strongly pan-WNK selective, inhibiting all four WNK isoforms. A class of quinoline compounds was identified that further shows selectivity among the WNK isoforms, being more potent toward WNK3 than WNK1. The crystal structure of the quinoline-derived SW120619 bound to the kinase domain of WNK3 reveals active site binding, and comparison to the WNK1 structure reveals the potential origin of isoform specificity.
Discussion:
The newly discovered classes of compounds may be starting points for generating pharmacological tools and potential drugs treating hypertension and cancer.
Insights
Researchers identified novel WNK kinase inhibitors, including quinoline compounds, from a large library screen. These inhibitors show pan-WNK selectivity and may lead to new treatments for hypertension and cancer.
Area of Science:
- Biochemistry
- Pharmacology
Background:
- WNK (with no lysine) kinases are serine/threonine kinases implicated in familial hyperkalemic hypertension (FHHt).
- WNK kinases represent therapeutic targets for managing blood pressure, stroke, and various cancers like triple-negative breast cancer and glioblastoma.
Purpose of the Study:
- To identify and characterize novel inhibitors targeting WNK kinases.
- To discover compounds with potential therapeutic applications in hypertension and cancer treatment.
Main Methods:
- A high-throughput screening of approximately 210,000 compounds was conducted.
- Assays included re-acquisition, specificity screening, and crystallography to identify WNK-isoform-selective inhibitors.
Main Results:
- Five distinct classes of WNK1 inhibitors were identified: quinoline compounds, halo-sulfones, cyclopropane-containing thiazoles, piperazine-containing compounds, and nitrophenol-derived compounds.
- All identified compounds demonstrated broad selectivity, inhibiting all four WNK isoforms (pan-WNK selectivity).
- A subset of quinoline compounds exhibited specific potency towards WNK3 over WNK1, with structural analysis revealing the basis for this selectivity.
Conclusions:
- The newly discovered compound classes serve as potential starting points for developing pharmacological tools.
- These inhibitors hold promise for the development of novel therapeutic agents for hypertension and various cancers.
