PS-1145 (IKK Inhibitor X) is a specific IKK inhibitor with IC50 of 88 nM.

IKKϵ:88 nM

50 mg/kg PS-1145 (i.v.)通过上调患有DMBA诱发的皮肤肿瘤的雄性Wistar大鼠中p53,激活胱天蛋白酶,并下调NF-κB和VEGF因子,增强肿瘤细胞凋亡.

PS-1145抑制人初级CD4(+) T细胞增殖,并通过CD3和CD28共同受体的参与阻碍响NF-κB 和 AP-1转录因子的活化。PS-1145通过抑制IkappaBalpha磷酸化阻断TNFalpha-诱导的NF-kappaB活化,并阻断IL-6对Dex-诱导的细胞凋亡的保护作用。

Kinase assay: PS-1145 are dissolved in DMSO and stored at ?20?°C until use. Ki value of PS-1145 against the IKK complex is determined by measuring Km ,?ATP against varying fixed concentration of the inhibitor. Briefly, partially purified IKK complex obtained from unstimulated HeLa S3 cells are pre-activated using the catalytic domain of MEKK1 expressed in sf9. Kinase activity is assessed using a biotinylated IκBα peptide (250 μM, RHDSGLDSMKD,Km ,peptide = 30 μM,K m ,?ATP = 10 μM) and phospho-[Ser32]-phosphoantibodies in an ELISA format with appropriate standard curve for quantification. For PS-1145 Ki measurement, the activated IKK complex is first preincubated in varying fixed concentration of the inhibitor (0.1–1 μM) at 25 °C for 1 h. Then apparentK m measurement for MgATP is performed at each discrete inhibitor concentration.

The inhibitory effect of PS-1145 on MM growth is assessed by measuring MTT dye absorbance of the cells. Cells from 48-h cultures are pulsed with 10 μL of 5 mg/ml MTT to each well for the last 4 h of 48-h cultures, followed by 100 μl of isopropanol containing 0.04N HCl. Absorbance is measured at 570 nm using a spectrophotometer. (Only for Reference)