title: Murine Efficacy and Pharmacokinetic Evaluation of the Flaviviral NS5 Capping Enzyme 2-Thioxothiazolidin-4-One Inhibitor BG-323.
abstract: Arthropod-borne flavivirus infection continues to cause significant morbidity and mortality worldwide. Identification of drug targets and novel antiflaviviral compounds to treat these diseases has become a global health imperative. A previous screen of 235,456 commercially available small molecules identified the 2-thioxothiazolidin-4-one family of compounds as inhibitors of the flaviviral NS5 capping enzyme, a promising target for antiviral drug development. Rational drug design methodologies enabled identification of lead compound BG-323 from this series. We have shown previously that BG-323 potently inhibits NS5 capping enzyme activity, displays antiviral effects in dengue virus replicon assays and inhibits growth of West Nile and yellow fever viruses with low cytotoxicity in vitro. In this study we further characterized BG-323's antiviral activity in vitro and in vivo. We found that BG-323 was able to reduce replication of WNV (NY99) and Powassan viruses in culture, and we were unable to force resistance into WNV (Kunjin) in long-term culture experiments. We then evaluated the antiviral activity of BG-323 in a murine model. Mice were challenged with WNV NY99 and administered BG-323 or mock by IP inoculation immediately post challenge and twice daily thereafter. Mice were bled and viremia was quantified on day three. No significant differences in viremia were observed between BG-323-treated and control groups and clinical scores indicated both BG-323-treated and control mice developed signs of illness on approximately the same day post challenge. To determine whether differences in in vitro and in vivo efficacy were due to unfavorable pharmacokinetic properties of BG-323, we conducted a pharmacokinetic evaluation of this small molecule. Insights from pharmacokinetic studies indicate that BG-323 is cell permeable, has a low efflux ratio and does not significantly inhibit two common cytochrome P450 (CYP P450) isoforms thus suggesting this molecule may be less likely to cause adverse drug interactions. However, the T1/2 of BG-323 was suboptimal and the percent of drug bound to plasma binding proteins was high. Future studies with BG-323 will be aimed at increasing the T1/2 and determining strategies for mitigating the effects of high plasma protein binding, which likely contribute to low in vivo efficacy.
authors.affiliation: University of Northern Colorado, School of Biological Sciences, Greeley, CO, United States of America.
authors.forename: Kristen M
authors.initials: KM
authors.lastname: Bullard
authors.affiliation: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, United States of America.
authors.forename: Rebekah C
authors.initials: RC
authors.lastname: Gullberg
authors.affiliation: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, United States of America.
authors.forename: Elnaz
authors.initials: E
authors.lastname: Soltani
authors.affiliation: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, United States of America.
authors.forename: J Jordan
authors.initials: JJ
authors.lastname: Steel
authors.affiliation: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, United States of America; Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, Colorado, United States of America.
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Geiss
authors.affiliation: University of Northern Colorado, School of Biological Sciences, Greeley, CO, United States of America.
authors.forename: Susan M
authors.initials: SM
authors.lastname: Keenan
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000493:pharmacokinetics* / Q000494:pharmacology*; D018938:Caco-2 Cells; D002522:Chlorocebus aethiops; D024882:Drug Resistance, Viral / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000493:pharmacokinetics* / Q000494:pharmacology*; D005260:Female; D006801:Humans; D051379:Mice; D008813:Mice, Inbred ICR; D011422:Propionates / Q000493:pharmacokinetics* / Q000494:pharmacology; D012315:RNA Caps / Q000737:chemistry / Q000378:metabolism*; D053778:Thiazolidines / Q000493:pharmacokinetics* / Q000494:pharmacology; D014018:Tissue Distribution; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors*; D014779:Virus Replication / Q000187:drug effects; D014901:West Nile Fever / Q000188:drug therapy* / Q000821:virology; D014902:West Nile virus / Q000187:drug effects*
chemical_list: C000607001:3-(5-(4-tert-butylbenzylidene-4-oxo-2-thioxo-1,3-thiazolidin-3-yl)propanoic acid; D000998:Antiviral Agents; D004791:Enzyme Inhibitors; C096041:NS5 protein, flavivirus; D011422:Propionates; D012315:RNA Caps; D053778:Thiazolidines; D017361:Viral Nonstructural Proteins