NR ARDE
AU Bertsch,U.; Winklhofer,K.F.; Hirschberger,T.; Bieschke,J.G.; Weber,P.; Hartl,F.U.; Tavan,P.; Tatzelt,J.; Kretzschmar,H.A.; Giese,A.
TI Systematic identification of anti-prion drugs by high-throughput screening based on scanning for intensely fluorescent targets (SIFT)
QU TSE-Forum, 4. Kongress - Nationale TSE-Forschungsplattform, Düsseldorf 28.10.-29.10.2004, Vortrag V-21
PT Konferenz-Vortrag
AB Conformational changes and aggregation of specific proteins are hallmarks of a number of neurodegenerative diseases, like Alzheimer's disease, Parkinson's disease and prion diseases. In the case of prion diseases the prion protein (PrP), a neuronal glycoprotein, undergoes a conformational change from the normal, mainly alpha-helical conformation to a disease-associated, mainly beta-sheeted, so called, scrapie-isoform (PrPsc), which forms amyloid aggregates. This conversion, which is crucial for disease progression, depends on direct PrPc/PrPsc interaction. We developed a high-throughput assay based on Scanning for Intensely Fluorescent Targets (SIFT) for the identification of drugs, which interfere with this interaction at the molecular level. Screening a library of 10,000 drug-like compounds yielded 256 primary hits, 80 of which were confirmed by dose-response curves with half-maximal inhibitory effects ranging from 0.3 to 60 µM. Among these, six compounds displayed an inhibitory effect on PrPsc propagation in scrapie-infected N2a cells. Four of these candidate drugs share a N'-benzylidene-benzohydrazide (NBB) core structure. Thus the combination of high-throughput in vitro assay with the established cell culture system provides a rapid and efficient method to identify new anti-prion drugs. Moreover, SIFT-based screening may facilitate the search for drugs against other diseases linked to protein aggregation.
AD Uwe Bertsch, Jan Bieschke, Petra Weber, Hans A. Kretzschmar, and Armin Giese, Zentrum für Neuropathologie und Prionforschung, Ludwig-Maximilians-Universität, Feodor-Lynen-Str. 23, D-81377 München, Germany; Konstanze F. Winklhofer, F. Ulrich Hartl, Jörg Tatzelt, Department of Cellular Biochemistry, Max-Planck-Institute for Biochemistry, Am Klopferspitz 18, D-82152 Martinsried, Germany; Thomas Hirschberger, Paul Tavan, Theoretische Biophysik, Lehrstuhl für BioMolekulare Optik, Ludwig-Maximilians-Universität, Oettingenstr. 67, D-80538 München, Germany; Jan Bieschke, Present address: The Scripps Research Institute, 10550 N. Torrey Pines Rd., BCC 265 La Jolla, CA 92037, USA
SP englisch
PO Deutschland
OR Tagungsband