Screening and identification of covalent Nrf2 activators with α,β-unsaturated imide structures

Area of Science:

• Biochemistry
• Molecular Biology
• Pharmacology

Background:

• The Keap1-Nrf2 pathway is crucial for cellular defense against oxidative stress.
• Pharmacological activation of this pathway offers therapeutic potential for neurodegenerative and inflammatory diseases.

Purpose of the Study:

• To screen for novel activators of the Keap1-Nrf2 pathway.
• To identify and characterize potent Nrf2 activators for therapeutic development.

Main Methods:

• High-throughput luciferase reporter assay for screening small molecules.
• Structure-activity relationship analysis of synthesized analogs.
• Cellular assays measuring glutathione levels and protection against oxidative damage.
• LC-MS analysis and covalent docking simulations to determine binding mechanisms.

Main Results:

• KU-0016, KU-0017, and KU-0171 were identified as moderate Nrf2 activators.
• KU-0479 demonstrated potent Nrf2 activation with low cytotoxicity.
• KU-0479 increased intracellular glutathione and protected cells from oxidative damage in a GSH-dependent manner.
• Evidence suggests KU-0479 covalently binds to Keap1 at the Cys151 residue.

Conclusions:

• KU-0479 is a synthetically accessible covalent Nrf2 activator.
• KU-0479 possesses favorable redox-modulating properties.
• KU-0479 exhibits significant therapeutic potential for oxidative stress-related diseases.