A high-throughput effector screen identifies a novel small molecule scaffold for inhibition of ten-eleven translocation dioxygenase 2.
Shubhendu PaleiJörn WeisnerMelina VogtRajesh GontlaBenjamin C BuchmullerChristiane EhrtTobias GrabeSilke KleinböltingMatthias Philipp MüllerGuido H CleverDaniel RauhDaniel SummererPublished in: RSC medicinal chemistry (2022)
Ten-eleven translocation dioxygenases (TETs) are the erasers of 5-methylcytosine (mC), the central epigenetic regulator of mammalian DNA. TETs convert mC to three oxidized derivatives with unique physicochemical properties and inherent regulatory potential, and it initializes active demethylation by the base excision repair pathway. Potent small molecule inhibitors would be useful tools to study TET functions by conditional control. To facilitate the discovery of such tools, we here report a high-throughput screening pipeline and its application to screen and validate 31.5k compounds for inhibition of TET2. Using a homogenous fluorescence assay, we discover a novel quinoline-based scaffold that we further validate with an orthogonal semi-high throughput MALDI-MS assay for direct monitoring of substrate turnover. Structure-activity relationship (SAR) studies involving >20 derivatives of this scaffold led to the identification of optimized inhibitors, and together with computational studies suggested a plausible model for its mode of action.
Keyphrases
- high throughput
- small molecule
- structure activity relationship
- tissue engineering
- mass spectrometry
- protein protein
- single cell
- single molecule
- case control
- transcription factor
- dna methylation
- multiple sclerosis
- gene expression
- circulating tumor
- dendritic cells
- ms ms
- genome wide
- cell free
- molecular docking
- bone mineral density
- climate change
- immune response
- anti inflammatory
- bioinformatics analysis
- nucleic acid
- postmenopausal women
- energy transfer