Discovery of a small-molecule inhibitor and cellular probe of Keap1-Nrf2 protein-protein interaction.
2013
Abstract A high-throughput screen (HTS) of the MLPCN library using a homogenous fluorescence polarization assay identified a small molecule as a first-in-class direct inhibitor of Keap1–Nrf2 protein–protein interaction. The HTS hit has three chiral centers; a combination of flash and chiral chromatographic separation demonstrated that Keap1-binding activity resides predominantly in one stereoisomer ( SRS )- 5 designated as ML334 (LH601A), which is at least 100× more potent than the other stereoisomers. The stereochemistry of the four cis isomers was assigned using X-ray crystallography and confirmed using stereospecific synthesis. ( SRS )- 5 is functionally active in both an ARE gene reporter assay and an Nrf2 nuclear translocation assay. The stereospecific nature of binding between ( SRS )- 5 and Keap1 as well as the preliminary but tractable structure–activity relationships support its use as a lead for our ongoing optimization
Keywords:
- Small molecule
- Protein–protein interaction
- Drug discovery
- Molecular probe
- Plasma protein binding
- Fluorescence anisotropy
- Stereochemistry
- Structure–activity relationship
- Small Molecule Libraries
- Chemistry
- Bardoxolone methyl
- Biochemistry
- Oxidative stress
- Signal transduction
- KEAP1
- Antioxidant Response Elements
- Oltipraz
- Correction
- Source
- Cite
- Save
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