Abstract
Prion diseases are associated with the conformational conversion of the physiological form of cellular prion protein (PrPC) to the pathogenic form, PrPSc. Compounds that inhibit this process by blocking conversion to the PrPSc could provide useful anti-prion therapies. However, no suitable drugs have been identified to date. To identify novel anti-prion compounds, we developed a combined structure- and ligand-based virtual screening system in silico. Virtual screening of a 700,000-compound database, followed by cluster analysis, identified 37 compounds with strong interactions with essential hotspot PrP residues identified in a previous study of PrPC interaction with a known anti-prion compound (GN8). These compounds were tested in vitro using a multimer detection system, cell-based assays, and surface plasmon resonance. Some compounds effectively reduced PrPSc levels and one of these compounds also showed a high binding affinity for PrPC. These results provide a promising starting point for the development of anti-prion compounds.
Highlights
Prion diseases are a group of lethal neurodegenerative diseases of humans and animals, including human Creutzfeldt-Jakob disease; bovine spongiform encephalopathy; scrapie in sheep, hamsters, and mice; and chronic wasting diseases in deer[1,2]
Anti-prion compounds are generally assessed by monitoring the levels of protease-resistant PrPSc using proteinase K (PK) digestion followed by western blotting
Several crystal and nuclear magnetic resonance (NMR) structures are available for PrPC, it should be noted that only the NMR of PrPC (1AG2) with GN8 structural details has been thoroughly characterised[23]
Summary
Prion diseases are a group of lethal neurodegenerative diseases of humans and animals, including human Creutzfeldt-Jakob disease; bovine spongiform encephalopathy; scrapie in sheep, hamsters, and mice; and chronic wasting diseases in deer[1,2]. The central event in prion disease pathogenesis is the conversion of the α -helix-rich cellular form of prion protein (PrPC) to a misfolded, β -sheet-rich, pathogenic, and infectious conformational isoform (PrPSc), the detailed structure of PrPSc is still not fully characterised[1,4,5] This conversion initiates a chain replication reaction, where each newly converted PrPSc molecule interacts with more PrPC molecules, fueling the formation of additional PrPSc6,7. Anti-prion compounds are generally assessed by monitoring the levels of protease-resistant PrPSc using proteinase K (PK) digestion followed by western blotting As this screening approach is fairly time-consuming and semi-quantitative, we employed a highly quantitative high-throughput misfolded protein detection assay (multimer detection system; MDS) to screen compounds for anti-prion efficacy. The T2 and 3E7 prion antibodies employed by the MDS recognize amino acids 147–152 and 140–160, respectively, of the PrP sequence[19]
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