Identification of potent Yes1 kinase inhibitors using a library screening approach

Abstract

Yes1 kinase has been implicated as a potential therapeutic target in a number of cancers including melanomas, breast cancers, and rhabdomyosarcomas. Described here is the development of a robust and miniaturized biochemical assay for Yes1 kinase that was applied in a high throughput screen (HTS) of kinase-focused small molecule libraries. The HTS provided 144 (17% hit rate) small molecule compounds with IC50 values in the sub-micromolar range. Three of the most potent Yes1 inhibitors were then examined in a cell-based assay for inhibition of cell survival in rhabdomyosarcoma cell lines. Homology models of Yes1 were generated in active and inactive conformations, and docking of inhibitors supports binding to the active conformation (DFG-in) of Yes1. This is the first report of a large high throughput enzymatic activity screen for identification of Yes1 kinase inhibitors, thereby elucidating the polypharmacology of a variety of small molecules and clinical candidates.

MEIS2 Is an Oncogenic Partner in AML1-ETO-Positive AML

“…Unfortunately, to date, no selective YES1 kinase inhibitor is available. Among the most potent YES1 inhibitors is dasatinib, which also blocks other src kinases (Patel et al, 2013). Although the data have to be interpreted with caution based on this, dasatinib a highly potent blocker of growth of AE-positive cells, and this was accompanied by dephosphorylation of the YES1 protein.…”

“…So far, there are no selective YES1 inhibitors available. One of the most potent YES1 inhibitors is dasatinib, which also impairs other kinases such as c-Src, Fyn, and Lyn (BMS-354825) (Patel et al, 2013). The AE-positive Kasumi cell line, expressing high levels of MEIS2, is also positive for YES1 expression and showed complete loss of phosphorylation of the kinase after dasatinib treatment.…”

Bioluminescence Methods for Assaying Kinases in Quantitative High-Throughput Screening (qHTS) Format Applied to Yes1 Tyrosine Kinase, Glucokinase, and PI5P4Kα Lipid Kinase

“…For the Yes1 assay, the average Z’= 0.76, CV= 6.9 and S/B =.23.7 (19). IC 50 for control dasatinib = 0.5 nM.…”

“…The ADP-Glo™ technology was applied to three classes of kinases here indicating the broad utility of this assay methodology: a lipid kinase PI5P4Kα (18), a tyrosine kinase Yes1 (19), and a metabolic kinase Glucokinase (20). The primary goal was to identify inhibitors of PI5P4Kα and Yes1 for the potential use as anticancer agents and activators of glucokinase for the potential treatment of diabetes.…”

Identification of Novel Plasmodium falciparum Hexokinase Inhibitors with Antiparasitic Activity

“…In step 1, 2 l of the ADP detection reagent was added and the plates were incubated for 40 min at RT. In step 2, 4 l of the kinase detection reagent was added and the plates were incubated for 30 min prior to reading of the luminescence on a ViewLux multimodal detector (PerkinElmer, Waltham, MA) using previously described methodologies (13)(14)(15)(16).…”