Therefore, it is very likely that the relevant molecular target of HAK compounds is involved in the OSM induced signal transduction process not earlier than 6 h after onset of the stimulation. Furthermore, IL 6 mRNA decay experiments were performed with actinomycin sellckchem D, a transcription arresting Inhibitors,Modulators,Libraries Inhibitors,Modulators,Libraries agent, to study whether the strong inhibition of IL 6 mRNA expression by HAK compounds Inhibitors,Modulators,Libraries was based on modified mRNA stability. No difference in mRNA stabi lity was observed between treated and non treated cells, demonstrating that the HAK com pound mediated suppression of IL 6 mRNA is most likely due to inhibition of transcription rather than modified mRNA stability.

Suppression of LPS induced Inhibitors,Modulators,Libraries IL 6 release by HAK compounds in primary murine astrocytes To analyze whether inhibition of OSM induced IL 6 expression is a cell line specific effect or a common fea ture of HAK compounds and valid in general, primary murine astrocytes were treated with HAK compounds. In contrast to human U343 glioma cells, OSM treatment did not lead to an increased IL 6 expression in mouse and rat primary astrocytes. However, LPS significantly induced IL 6 release into the condi tioned medium of mouse and rat astrocyte cultures. Primary murine astrocytes stimulated with 1 ug ml LPS were co treated with compounds HAK 2 and HAK 5 at a concentration of 20 uM each for 24 h. The observed effect of compounds HAK 2 and HAK 5 on LPS stimulation was similar to that of OSM induced IL 6 expression in human U343 glioma cells. In comparison to untreated samples LPS induced IL 6 expression was reduced by 60% in mouse and 50% in rat astrocytes by both HAK compounds.

Thus, suppression of both, LPS and OSM induced IL 6 expression in different cell types Inhibitors,Modulators,Libraries by structurally related compounds is another indication for strong potency of HAK compounds to target neuroinflammatory processes. Potent inhibition of IL 6 upregulation by compound HAK 2 in vivo Based on the results obtained from primary murine astrocytes, we reasoned that HAK compounds might also suppress elevated IL 6 expression in vivo. There fore, bioactivity of the compound HAK 2 was analyzed in the LPS induced mouse septic shock model. In pre paration of this in vivo study, selected HAK compounds were characterized in detail concerning brain bioavail ability.

It was www.selleckchem.com/products/Enzastaurin.html demonstrated by quantitative analysis of mouse plasma and brain samples using LC MS MS that the compounds are bioavailable and able to pass the blood brain barrier. For further in vivo investigations compound HAK 2 with a logBB of 0. 22 was selected. Intraperitoneal injection of 1 mg kg LPS into C57 B6 mice resulted in an acute elevation of IL 6 concentra tion in plasma, hippocampus and cortex 2 h post administration. The plasma level of IL 6 protein was significantly reduced by 55% in mice treated with 5 mg kg HAK 2 in parallel as compared to LPS alone.