TBK1/IKKε-IN-4 is a 6-aminopyrazolopyrimidine derivative and a potent, selective TBK1 and IKKε inhibitor with IC 50 values of 13 nM and 59 nM, respectively. TBK1/IKKε-IN-4 shows 100- to 1000-fold less activity against other protein kinases including PDK1, PI3K family members and mTOR In Vitro TBK1/IKKε-IN-4 (Compound II; 96 hours; A549 andHCC44 cells) treatmentdisplays selective toxicity in TBK1-dependent cancer cell lines (IC 50 of ~ 4.2 µM for H441 cells and IC 50 of ~0.4 µM for A549 cells). TBK1/IKKε-IN-4 (Compound II; 0-2 µM; 30 minutes; HCC44 cells) treatment inhibits the AKT activity. TBK1/IKKε-IN-4 (Compound II) inhibits LPS-induced expression of IFNβ (IC 50 =62 nM), and the IFNβ target genes IP10 (IC 50 =78 nM) and Mx1 (IC 50 =20 nM). TBK1/IKKε-IN-4 effectively blocksTLR3-dependent IRF3 nuclear translocation in cells with an IC 50 under 100 nM, but does not impair TNFR1-dependent p65 NFκB nuclear translocation with doses as high as 20 µM. MCE has not independently confirmed the accuracy of these methods. They are for reference only. Western Blot AnalysisCell Line: HCC44 cells Concentration: 0 µM, 0.5 µM, 1 µM, 2 µM Incubation Time: 30 minutes Result: Was sufficient to blunt baseline AKT activity. Form:Solid IC50& Target:IKKε 59 nM (IC 50 ) TBK1 13 nM (IC 50 )