Structural, kinetic, and pharmacodynamic mechanisms of D-amino acid oxidase inhibition by small molecules

J Med Chem. 2013 May 9;56(9):3710-24. doi: 10.1021/jm4002583. Epub 2013 Apr 30.


We characterized the mechanism and pharmacodynamics of five structurally distinct inhibitors of d-amino acid oxidase. All inhibitors bound the oxidized form of human enzyme with affinity slightly higher than that of benzoate (Kd ≈ 2-4 μM). Stopped-flow experiments showed that pyrrole-based inhibitors possessed high affinity (Kd ≈ 100-200 nM) and slow release kinetics (k < 0.01 s(-1)) in the presence of substrate, while inhibitors with pendent aromatic groups altered conformations of the active site lid, as evidenced by X-ray crystallography, and showed slower kinetics of association. Rigid bioisosteres of benzoic acid induced a closed-lid conformation, had slower release in the presence of substrate, and were more potent than benzoate. Steady-state d-serine concentrations were described in a PK/PD model, and competition for d-serine sites on NMDA receptors was demonstrated in vivo. DAAO inhibition increased the spatiotemporal influence of glial-derived d-serine, suggesting localized effects on neuronal circuits where DAAO can exert a neuromodulatory role.

MeSH terms

  • Animals
  • Behavior, Animal / drug effects
  • Binding, Competitive
  • Catalytic Domain
  • D-Amino-Acid Oxidase / antagonists & inhibitors*
  • D-Amino-Acid Oxidase / chemistry
  • D-Amino-Acid Oxidase / metabolism
  • Enzyme Inhibitors / chemistry*
  • Enzyme Inhibitors / metabolism
  • Enzyme Inhibitors / pharmacokinetics
  • Enzyme Inhibitors / pharmacology*
  • Glycine / metabolism
  • Humans
  • Kinetics
  • Male
  • Molecular Docking Simulation
  • Neuroglia / drug effects
  • Neuroglia / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Serine / biosynthesis
  • Small Molecule Libraries / chemistry*
  • Small Molecule Libraries / metabolism
  • Small Molecule Libraries / pharmacokinetics
  • Small Molecule Libraries / pharmacology*


  • Enzyme Inhibitors
  • Small Molecule Libraries
  • Serine
  • D-Amino-Acid Oxidase
  • Glycine

Associated data

  • PDB/3ZNN
  • PDB/3ZNO
  • PDB/3ZNP
  • PDB/3ZNQ