Asciminib

All compounds (imatinib, nilotinib, dasatinib, ponatinib, crizotinib, and asciminib) used in this study were purchased from Selleck Chemicals (Houston, TX, USA) and were diluted in DMSO to a 1000× stock solution, which then were diluted to 1× working concentrations for the experiments. For the in vivo experiments, the compounds were diluted to working concentration in 0.5% methyl-cellulose (Methocel® 65HG, Fluka, Darmstadt, Germany).

Asciminib, (ASCI), bosutinib (BOSU), dasatinib (DASA), imatinib (IMA), nilotinib (NILO) and ponatinib (PONA) were obtained from Selleckchem (Cologne, Germany). The TKI were dissolved in DMSO and stored at −80 °C. Stock concentrations were 5 mM for ASCI, BOSU, DASA and PONA, and 10 mM for IMA and NILO. Peak and IC50 concentrations were chosen based on clinical studies which reported serum levels in patient samples. Peak levels as well as steady-state concentrations (from here on: IC50) were based on these data [20 (link),21 (link),22 (link),23 (link),24 (link),25 (link),26 (link),27 (link)]. The concentrations were chosen as follows: ASCI 806 nM (peak) and 28 nM (IC50); BOSU 600 nM (peak) and 2.4 nM (IC50); DASA 150 nM (peak) and 10 nM (IC50); IMA 3000 nM (peak) and 600 nM (IC50); NILO 3000 nM (peak) and 30 nM (IC50); PONA 145 nM (peak) and 70 nM (IC50). DMSO was included as a control in all experiments at a maximum concentration of 0.1% v/v in NILO peak samples.

KIRA8 (CAS#1630086-20-2), GNF-2 (CAS#778270-11-4), and thapsigargin (CAS#67526-95-8) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Asciminib (CAS#1492952-76-7) was purchased from Selleck Chemicals (Houston, TX, USA). All reagents were dissolved in dimethyl sulfoxide (DMSO) to a final concentration of 10 mM as a stock solution and stored at −30 °C.