Discovery and Characterization of a Chemical Probe for Cyclin-Dependent Kinase-Like 2.
Frances M BashoreSophia M MinXiangrong ChenStefanie HowellCaroline H RinderleGabriel MorelJosie A SilvaroliCarrow I WellsBruce A BunnellDavid Harold DrewryNavjot S PablaSila K UltanirAlex N BullockAlison D AxtmanPublished in: ACS medicinal chemistry letters (2024)
Acylaminoindazole-based inhibitors of CDKL2 were identified via analyses of cell-free binding and selectivity data. Compound 9 was selected as a CDKL2 chemical probe based on its potent inhibition of CDKL2 enzymatic activity, engagement of CDKL2 in cells, and excellent kinome-wide selectivity, especially when used in cells. Compound 16 was designed as a negative control to be used alongside compound 9 in experiments to interrogate CDKL2-mediated biology. A solved cocrystal structure of compound 9 bound to CDKL2 highlighted key interactions it makes within its ATP-binding site. Inhibition of downstream phosphorylation of EB2, a CDKL2 substrate, in rat primary neurons provided evidence that engagement of CDKL2 by compound 9 in cells resulted in inhibition of its activity. When used at relevant concentrations, compound 9 does not impact the viability of rat primary neurons or certain breast cancer cells nor elicit consistent changes in the expression of proteins involved in epithelial-mesenchymal transition.
Keyphrases
- induced apoptosis
- cell cycle arrest
- epithelial mesenchymal transition
- cell free
- oxidative stress
- signaling pathway
- spinal cord
- cell death
- social media
- poor prognosis
- breast cancer cells
- quantum dots
- small molecule
- cell cycle
- transcription factor
- deep learning
- hydrogen peroxide
- spinal cord injury
- binding protein
- transforming growth factor
- big data
- amino acid