NR ASOA
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 antiprion drugs by high-throughput screening based on scanning for intensely fluorescent targets
QU Journal of Virology 2005 Jun; 79(12): 7785-91
IA http://jvi.asm.org/cgi/content/full/79/12/7785?view=long&pmid=15919931
PT journal article
AB Conformational changes and aggregation of specific proteins are hallmarks of a number of 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 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 of 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 microM. Among these, six compounds displayed an inhibitory effect on PrPsc propagation in scrapie-infected N2a cells. Four of these candidate drugs share an N'-benzylidene-benzohydrazide 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 antiprion drugs, which corroborates that interaction of PrPc and PrPsc is a crucial molecular step in the propagation of prions. Moreover, SIFT-based screening may facilitate the search for drugs against other diseases linked to protein aggregation.
MH Animals; Benzylidene Compounds/chemistry/*pharmacology; Cell Line; Fluorescence; Humans; Mice; *Pharmaceutical Preparations/chemistry; PrPc Proteins/chemistry; PrPsc Proteins/chemistry; Prion Diseases/drug therapy; Prions/*antagonists & inhibitors/chemistry; Protein Binding; Protein Conformation; Protein Folding; Protein Interaction Mapping/*methods; Research Support, Non-U.S. Gov't
AD Zentrum für Neuropathologie und Prionforschung, Ludwig Maximilians Universität, Feodor Lynen Str. 23, D-81377 München, Germany.
SP englisch
PO USA