title: Inhibition of Heat-Stable Toxin-Induced Intestinal Salt and Water Secretion by a Novel Class of Guanylyl Cyclase C Inhibitors.
abstract: BACKGROUND
  Many enterotoxigenic Escherichia coli strains produce the heat-stable toxin, STa, which, by activation of the intestinal receptor-enzyme guanylyl cyclase (GC) C, triggers an acute, watery diarrhea. We set out to identify GCC inhibitors that may be of benefit for the treatment of infectious diarrheal disease.
  
  
  METHODS
  Compounds that inhibit STa-induced cyclic guanosine 3',5'-monophosphate (cGMP) production were selected by performing cyclase assays on cells and membranes containing GCC, or the related GCA. The effect of leads on STa/GCC-dependent activation of the cystic fibrosis transmembrane conductance regulator anion channel was investigated in T84 cells, and in porcine and human intestinal tissue. Their effect on STa-provoked fluid transport was assessed in ligated intestinal loops in piglets.
  
  
  RESULTS
  Four N-2-(propylamino)-6-phenylpyrimidin-4-one-substituted piperidines were shown to inhibit GCC-mediated cellular cGMP production. The half maximal inhibitory concentrations were ≤ 5 × 10(-7) mol/L, whereas they were >10 times higher for GCA. In T84 monolayers, these leads blocked STa/GCC-dependent, but not forskolin/adenylyl cyclase-dependent, cystic fibrosis transmembrane conductance regulator activity. GCC inhibition reduced STa-provoked anion secretion in pig jejunal tissue, and fluid retention and cGMP levels in STa-exposed loops. These GCC inhibitors blocked STa-provoked anion secretion in rectal biopsy specimens.
  
  
  CONCLUSIONS
  We have identified a novel class of GCC inhibitors that may form the basis for development of future therapeutics for (infectious) diarrheal disease.
authors.affiliation: Department of Gastroenterology and Hepatology, Erasmus MC University Medical Center, Rotterdam, The Netherlands.
authors.forename: Marcel J C
authors.initials: MJ
authors.lastname: Bijvelds
authors.affiliation: Laboratory of Immunology, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
authors.forename: Michaela
authors.initials: M
authors.lastname: Loos
authors.affiliation: Department of Pulmonology, University Medical Center Utrecht, The Netherlands.
authors.forename: Inez
authors.initials: I
authors.lastname: Bronsveld
authors.affiliation: Shire-Movetis NV, Turnhout.
authors.forename: Ann
authors.initials: A
authors.lastname: Hellemans
authors.affiliation: Janssen Research and Development, Janssen Pharmaceutica NV, Beerse, Belgium.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Bongartz
authors.affiliation: Janssen Research and Development, Janssen Pharmaceutica NV, Beerse, Belgium.
authors.forename: Luc
authors.initials: L
authors.lastname: Ver Donck
authors.affiliation: Laboratory of Immunology, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
authors.forename: Eric
authors.initials: E
authors.lastname: Cox
authors.affiliation: Department of Gastroenterology and Hepatology, Erasmus MC University Medical Center, Rotterdam, The Netherlands.
authors.forename: Hugo R
authors.initials: HR
authors.lastname: de Jonge
authors.affiliation: Shire-Movetis NV, Turnhout.
authors.forename: Jan A J
authors.initials: JA
authors.lastname: Schuurkes
authors.affiliation: Shire-Movetis NV, Turnhout.
authors.forename: Joris H
authors.initials: JH
authors.lastname: De Maeyer
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism; D000328:Adult; D000818:Animals; D001427:Bacterial Toxins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000037:antagonists & inhibitors / Q000378:metabolism; D003967:Diarrhea; D054307:Enterotoxigenic Escherichia coli; D004768:Enterotoxins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D029968:Escherichia coli Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007583:Jejunum / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008954:Models, Biological; D010880:Piperidines / Q000494:pharmacology*; D000074261:Receptors, Enterotoxin; D050705:Receptors, Guanylate Cyclase-Coupled / Q000037:antagonists & inhibitors* / Q000378:metabolism; D018000:Receptors, Peptide / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013552:Swine; D055815:Young Adult
chemical_list: D001427:Bacterial Toxins; D004768:Enterotoxins; D004791:Enzyme Inhibitors; D029968:Escherichia coli Proteins; D010880:Piperidines; D018000:Receptors, Peptide; C034151:heat stable toxin (E coli); D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000262:Adenylyl Cyclases; D000074261:Receptors, Enterotoxin; D050705:Receptors, Guanylate Cyclase-Coupled
keywords: CFTR; enterotoxigenic E. coli (ETEC); guanylyl cyclase C; secretory diarrhea