Natural products AKTing on signal transduction pathways-molecules and methods

摘要: Natural products play an important role as source of inspiration for drug discovery and development and tools basic research. Isolated natural that had shown diverse biological activity in previously published tests were analysed their direct effect on molecular signalling processes, militarinone A from the entomogenous fungus Paecilomyces militaris indolinone Isatis tinctoria. The fungal alkaloid was originally identified a screening for neurotrophic substances, where it induced neuronal outgrowth PC12 cells. To uncover the mechanism this action we studied cell pathways involved spike formation differentiation two types cells (PC12 N2a) interaction with associated pathways. The increased could not be confirmed general of militarinone A, only transiently seen cells all other cell lines tested underwent apoptosis within 24h. We propose difference is due to varying constitutive levels p53. Furthermore, traditional European medicinal plant tinctoria was analysed. tinctoria contains several known anti-inflammatory components, namely, tryptanthrin, indirubin, indolinone. In previous study, was shown to inhibit degranulation mast anti-allergic indolinone should further characterised. initially observed stabilising effect on different species assay set-up showed that indolinone efficiently blocked PtdInsP3 production due inhibition class I PI3- kinases, therefore preventing activation Akt subsequent cell degranulation. The concentrations necessary obtain vitro, however, too high consider vivo testing. Since depends phosphoinositide studied phosphosinositide upon stimulation. For purpose, developed a method allows individual analysis phosphoinositides, including PtdInsPand PtdInsP2-isomers. This novel method, based ion-pair chromatography and ESI-MS detection, offers substantial perspectives application phosphoinositidesignalling research relative quantification PIs