title: Inhibitory effect of REP3123 on toxin and spore formation in Clostridium difficile, and in vivo efficacy in a hamster gastrointestinal infection model.
abstract: OBJECTIVE
  REP3123 is a fully synthetic methionyl-tRNA synthetase inhibitor in pre-clinical development as a novel agent to treat Clostridium difficile infection (CDI). This novel agent was investigated for its ability to block the production of toxins and spores, and was tested for efficacy in vivo in a hamster model.
  
  
  METHODS
  Clostridial toxin levels were determined qualitatively using monoclonal antibodies and by cytotoxicity assays. Spores were detected by staining and by quantitative dilution plating after ethanol treatment. Efficacy of REP3123 was tested in a clindamycin-induced C. difficile hamster gastrointestinal (GI) infection model.
  
  
  RESULTS
  REP3123 at concentrations as low as 1 mg/L inhibited de novo toxin production in high cell density, stationary phase cultures of C. difficile. Among comparator agents currently used for CDI therapy, vancomycin required much higher levels of 20 mg/L, and metronidazole had no effect on toxin levels. REP3123 caused a >10-fold reduction of the sporulation rate in vitro. Vancomycin and, in particular, metronidazole appeared to promote the formation of spores. REP3123, at concentrations as low as 0.5 mg/kg, demonstrated efficacy in the hamster model of CDI and was superior to vancomycin in the overall survival of the animals at the end of the study (33 days).
  
  
  CONCLUSIONS
  REP3123 inhibited growth of C. difficile, affected the production of toxins and spores and demonstrated superior efficacy compared with vancomycin in the hamster GI infection model. This agent may be a promising candidate for CDI treatment; in particular, the inhibition of toxin production and spore formation may reduce the severity and spread of the disease, respectively.
authors.affiliation: Replidyne, Inc., Louisville, CO 80027, USA. ursochsner@gmail.com
authors.forename: Urs A
authors.initials: UA
authors.lastname: Ochsner
authors.forename: Stacie J
authors.initials: SJ
authors.lastname: Bell
authors.forename: Ann L
authors.initials: AL
authors.lastname: O'Leary
authors.forename: Teresa
authors.initials: T
authors.lastname: Hoang
authors.forename: Kimberley Clawson
authors.initials: KC
authors.lastname: Stone
authors.forename: Casey L
authors.initials: CL
authors.lastname: Young
authors.forename: Ian A
authors.initials: IA
authors.lastname: Critchley
authors.forename: Nebojsa
authors.initials: N
authors.lastname: Janjic
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology / Q000627:therapeutic use*; D001427:Bacterial Toxins / Q000096:biosynthesis*; D001578:Benzopyrans / Q000494:pharmacology*; D002460:Cell Line; D016360:Clostridioides difficile / Q000187:drug effects*; D006224:Cricetinae; D004761:Enterocolitis, Pseudomembranous / Q000188:drug therapy*; D004791:Enzyme Inhibitors / Q000494:pharmacology / Q000627:therapeutic use*; D008297:Male; D013171:Spores, Bacterial / Q000187:drug effects*; D016019:Survival Analysis; D013876:Thiophenes / Q000494:pharmacology*; D014640:Vancomycin / Q000494:pharmacology / Q000627:therapeutic use
chemical_list: D000900:Anti-Bacterial Agents; D001427:Bacterial Toxins; D001578:Benzopyrans; D004791:Enzyme Inhibitors; C541771:REP 3123; D013876:Thiophenes; D014640:Vancomycin