title: Small-molecule activators of insulin-degrading enzyme discovered through high-throughput compound screening.
abstract: BACKGROUND
  Hypocatabolism of the amyloid beta-protein (Abeta) by insulin-degrading enzyme (IDE) is implicated in the pathogenesis of Alzheimer disease (AD), making pharmacological activation of IDE an attractive therapeutic strategy. However, it has not been established whether the proteolytic activity of IDE can be enhanced by drug-like compounds.
  
  
  RESULTS
  Based on the finding that ATP and other nucleotide polyphosphates modulate IDE activity at physiological concentrations, we conducted parallel high-throughput screening campaigns in the absence or presence of ATP and identified two compounds--designated Ia1 and Ia2--that significantly stimulate IDE proteolytic activity. Both compounds were found to interfere with the crosslinking of a photoaffinity ATP analogue to IDE, suggesting that they interact with a bona fide ATP-binding domain within IDE. Unexpectedly, we observed highly synergistic activation effects when the activity of Ia1 or Ia2 was tested in the presence of ATP, a finding that has implications for the mechanisms underlying ATP-mediated activation of IDE. Notably, Ia1 and Ia2 activated the degradation of Abeta by approximately 700% and approximately 400%, respectively, albeit only when Abeta was presented in a mixture also containing shorter substrates.
  
  
  CONCLUSIONS
  This study describes the first examples of synthetic small-molecule activators of IDE, showing that pharmacological activation of this important protease with drug-like compounds is achievable. These novel activators help to establish the putative ATP-binding domain as a key modulator of IDE proteolytic activity and offer new insights into the modulatory action of ATP. Several larger lessons abstracted from this screen will help inform the design of future screening campaigns and facilitate the eventual development of IDE activators with therapeutic utility.
authors.affiliation: Department of Neuroscience, Mayo Clinic, Jacksonville, Florida, United States of America.
authors.forename: Christelle
authors.initials: C
authors.lastname: Cabrol
authors.forename: Malwina A
authors.initials: MA
authors.lastname: Huzarska
authors.forename: Christopher
authors.initials: C
authors.lastname: Dinolfo
authors.forename: Maria C
authors.initials: MC
authors.lastname: Rodriguez
authors.forename: Lael
authors.initials: L
authors.lastname: Reinstatler
authors.forename: Jake
authors.initials: J
authors.lastname: Ni
authors.forename: Li-An
authors.initials: LA
authors.lastname: Yeh
authors.forename: Gregory D
authors.initials: GD
authors.lastname: Cuny
authors.forename: Ross L
authors.initials: RL
authors.lastname: Stein
authors.forename: Dennis J
authors.initials: DJ
authors.lastname: Selkoe
authors.forename: Malcolm A
authors.initials: MA
authors.lastname: Leissring
mesh_terms: D016229:Amyloid beta-Peptides / Q000378:metabolism; D002626:Chemistry, Pharmaceutical; D020536:Enzyme Activators / Q000494:pharmacology*; D006868:Hydrolysis; D007328:Insulin / Q000378:metabolism*; D007339:Insulysin / Q000378:metabolism*; D019920:Photoaffinity Labels
chemical_list: D016229:Amyloid beta-Peptides; D020536:Enzyme Activators; D007328:Insulin; D019920:Photoaffinity Labels; D007339:Insulysin