title: Lifting the mask: identification of new small molecule inhibitors of uropathogenic Escherichia coli group 2 capsule biogenesis.
abstract: Uropathogenic Escherichia coli (UPEC) is the leading cause of community-acquired urinary tract infections (UTIs), with over 100 million UTIs occurring annually throughout the world. Increasing antimicrobial resistance among UPEC limits ambulatory care options, delays effective treatment, and may increase overall morbidity and mortality from complications such as urosepsis. The polysaccharide capsules of UPEC are an attractive target a therapeutic, based on their importance in defense against the host immune responses; however, the large number of antigenic types has limited their incorporation into vaccine development. The objective of this study was to identify small-molecule inhibitors of UPEC capsule biogenesis. A large-scale screening effort entailing 338,740 compounds was conducted in a cell-based, phenotypic screen for inhibition of capsule biogenesis in UPEC. The primary and concentration-response assays yielded 29 putative inhibitors of capsule biogenesis, of which 6 were selected for further studies. Secondary confirmatory assays identified two highly active agents, named DU003 and DU011, with 50% inhibitory concentrations of 1.0 µM and 0.69 µM, respectively. Confirmatory assays for capsular antigen and biochemical measurement of capsular sugars verified the inhibitory action of both compounds and demonstrated minimal toxicity and off-target effects. Serum sensitivity assays demonstrated that both compounds produced significant bacterial death upon exposure to active human serum. DU011 administration in mice provided near complete protection against a lethal systemic infection with the prototypic UPEC K1 isolate UTI89. This work has provided a conceptually new class of molecules to combat UPEC infection, and future studies will establish the molecular basis for their action along with efficacy in UTI and other UPEC infections.
authors.affiliation: Department. of Pediatrics, Duke University School of Medicine, Durham, North Carolina, United States of America.
authors.forename: Carlos C
authors.initials: CC
authors.lastname: Goller
authors.affiliation: Department. of Pediatrics, Duke University School of Medicine, Durham, North Carolina, United States of America.
authors.forename: Mehreen
authors.initials: M
authors.lastname: Arshad
authors.affiliation: Southern Research Specialized Biocontainment Screening Center, Southern Research Institute, Birmingham, Alabama, United States of America.
authors.forename: James W
authors.initials: JW
authors.lastname: Noah
authors.affiliation: Southern Research Specialized Biocontainment Screening Center, Southern Research Institute, Birmingham, Alabama, United States of America.
authors.forename: Subramaniam
authors.initials: S
authors.lastname: Ananthan
authors.affiliation: Southern Research Specialized Biocontainment Screening Center, Southern Research Institute, Birmingham, Alabama, United States of America.
authors.forename: Carrie W
authors.initials: CW
authors.lastname: Evans
authors.affiliation: Southern Research Specialized Biocontainment Screening Center, Southern Research Institute, Birmingham, Alabama, United States of America.
authors.forename: N Miranda
authors.initials: NM
authors.lastname: Nebane
authors.affiliation: Southern Research Specialized Biocontainment Screening Center, Southern Research Institute, Birmingham, Alabama, United States of America.
authors.forename: Lynn
authors.initials: L
authors.lastname: Rasmussen
authors.affiliation: Southern Research Specialized Biocontainment Screening Center, Southern Research Institute, Birmingham, Alabama, United States of America.
authors.forename: Melinda
authors.initials: M
authors.lastname: Sosa
authors.affiliation: Southern Research Specialized Biocontainment Screening Center, Southern Research Institute, Birmingham, Alabama, United States of America.
authors.forename: Nichole A
authors.initials: NA
authors.lastname: Tower
authors.affiliation: Southern Research Specialized Biocontainment Screening Center, Southern Research Institute, Birmingham, Alabama, United States of America.
authors.forename: E Lucile
authors.initials: EL
authors.lastname: White
authors.affiliation: Specialized Chemistry Center, University of Kansas, Lawrence, Kansas, United States of America.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Neuenswander
authors.affiliation: Specialized Chemistry Center, University of Kansas, Lawrence, Kansas, United States of America.
authors.forename: Patrick
authors.initials: P
authors.lastname: Porubsky
authors.affiliation: Specialized Chemistry Center, University of Kansas, Lawrence, Kansas, United States of America.
authors.forename: Brooks E
authors.initials: BE
authors.lastname: Maki
authors.affiliation: Specialized Chemistry Center, University of Kansas, Lawrence, Kansas, United States of America.
authors.forename: Steven A
authors.initials: SA
authors.lastname: Rogers
authors.affiliation: Specialized Chemistry Center, University of Kansas, Lawrence, Kansas, United States of America.
authors.forename: Frank
authors.initials: F
authors.lastname: Schoenen
authors.affiliation: Department. of Pediatrics, Duke University School of Medicine, Durham, North Carolina, United States of America; Department of Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, North Carolina, United States of America; Center for Microbial Pathogenesis, Duke University School of Medicine, Durham, North Carolina, United States of America.
authors.forename: Patrick C
authors.initials: PC
authors.lastname: Seed
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology; D016667:Bacterial Capsules / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D004927:Escherichia coli Infections / Q000188:drug therapy* / Q000473:pathology; D005260:Female; D006801:Humans; D051379:Mice; D014552:Urinary Tract Infections / Q000188:drug therapy* / Q000473:pathology; D056689:Uropathogenic Escherichia coli / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents