title: Antiprion Activity of DB772 and Related Monothiophene- and Furan-Based Analogs in a Persistently Infected Ovine Microglia Culture System.
abstract: The transmissible spongiform encephalopathies are fatal neurodegenerative disorders characterized by the misfolding of the native cellular prion protein (PrP(C)) into the accumulating, disease-associated isoform (PrP(Sc)). Despite extensive research into the inhibition of prion accumulation, no effective treatment exists. Previously, we demonstrated the inhibitory activity of DB772, a monocationic phenyl-furan-benzimidazole, against PrP(Sc) accumulation in sheep microglial cells. In an effort to determine the effect of structural substitutions on the antiprion activity of DB772, we employed an in vitro strategy to survey a library of structurally related, monothiophene- and furan-based compounds for improved inhibitory activity. Eighty-nine compounds were screened at 1 μM for effects on cell viability and prion accumulation in a persistently infected ovine microglia culture system. Eleven compounds with activity equivalent to or higher than that of DB772 were identified as preliminary hit compounds. For the preliminary hits, cytotoxicities and antiprion activities were compared to calculate the tissue culture selectivity index. A structure-activity relationship (SAR) analysis was performed to determine molecular components contributing to antiprion activity. To investigate potential mechanisms of inhibition, effects on PrP(C) and PrP(Sc) were examined. While inhibition of total PrP(C) was not observed, the results suggest that a potential target for inhibition at biologically relevant concentrations is through PrP(C) misfolding to PrP(Sc) Further, SAR analysis suggests that two structural elements were associated with micromolar antiprion activity. Taken together, the described data provide a foundation for deeper investigation into untested DB compounds and in the design of effective therapeutics.
authors.affiliation: Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington, USA.
authors.forename: Kelcey D
authors.initials: KD
authors.lastname: Dinkel
authors.affiliation: Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington, USA Department of Pathology, University of Georgia, Athens, Georgia, USA.
authors.forename: James B
authors.initials: JB
authors.lastname: Stanton
authors.affiliation: Department of Chemistry, Georgia State University, Atlanta, Georgia, USA.
authors.forename: David W
authors.initials: DW
authors.lastname: Boykin
authors.affiliation: Department of Chemistry and Physics, Augusta University, Augusta, Georgia, USA.
authors.forename: Chad E
authors.initials: CE
authors.lastname: Stephens
authors.affiliation: Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington, USA.
authors.forename: Sally A
authors.initials: SA
authors.lastname: Madsen-Bouterse
authors.affiliation: Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington, USA United States Department of Agriculture, Agricultural Research Service, Animal Disease Research Unit, Pullman, Washington, USA david.schneider1@ars.usda.gov.
authors.forename: David A
authors.initials: DA
authors.lastname: Schneider
mesh_terms: D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology*; D005663:Furans / Q000494:pharmacology*; D017628:Microglia / Q000187:drug effects*; D018620:PrPSc Proteins / Q000037:antagonists & inhibitors*; D000072002:Prion Proteins / Q000037:antagonists & inhibitors*; D012756:Sheep; D013329:Structure-Activity Relationship
chemical_list: C571880:2-(2-benzimidazolyl)-5-(4-(2-imidazolino)phenyl)furan; D001562:Benzimidazoles; D005663:Furans; D018620:PrPSc Proteins; D000072002:Prion Proteins; C031000:benzimidazole