Bay 11-7085 can irreversibly inhibit the IκBα phosphorylation induced by TNFα (IC50: 10 μM).

IκBα phosphorylation:10 μM

在含Caov-3细胞的无胸腺裸鼠体内,BAY 11-7085可使紫杉醇对腹内传播和腹水产生的抑制作用增强.BAY11-7085对脑膜炎相关的NF-κB活性增加有抑制作用,改善被感染大鼠临床状态,并使脑膜炎相关的CNS并发症和脑膜炎症显著衰减.

BAY 11-7085与紫杉醇联用可增强对NF-κB活性的抑制,并使经紫杉醇处理细胞的存活率降低。通过抑制NF-κB,BAY11-7085可抑制TNFα诱导的粘附分子E-selectin,ICAM-1及VCAM-1的表达。BAY11-7085（10 μM）对HUVEC细胞无细胞毒性。此外,Bay11-7085与LY294002联用可协同影响PEL细胞的凋亡。

In Gel Kinase Assay: In gel kinase assay for the proteins that phosphorylate IκB-α is carried out as detailed below. Whole cell extracts are prepared from HUVEC treated with TNFα (100 units/ml) for 15 min in the presence or absence of inhibitor (20 μM, pretreatment for 1 h) as indicated. Proteins are separated on a 10% SDS gel containing 0.5 mg/ml HIS-IκB-α. Gels are washed two times in 20% propanol, 50 mM Hepes, pH 7.6, for 30 min and two times in buffer A (50 mM Hepes, pH 7.6, 5 mM 2-mercaptoethanol) for 30 min, followed by a 1-h incubation with buffer A containing 6 M urea, 1 h each in 3, 1.5, and 0.75 M urea in buffer A and 0.05% Tween 20 and 1 h in buffer A with 0.05% Tween 20. The kinase assay is carried out for 1 h at 30?°C in the presence of 50 μM ATP, 5 μCi/ml [32P]ATP, 20 mM Hepes, pH 7.6, 20 mM MgCl2, 20 mM β-glycerophosphate, 20 mM p-nitrophenyl phosphate, 1 mM sodium vanadate, 2 mM dithiothreitol. The gel is washed with 5% trichloroacetic acid and 1% sodium pyrophosphate, dried, and exposed to film. A separate gel with no HIS-IκB-α is assayed as a control.

Human umbilical vein endothelial cells (HUVEC) are isolated and maintained in culture. Cell toxicity is assessed by morphology and by MTT assay.(Only for Reference)