4 research outputs found
An NMR Biochemical Assay for Fragment-Based Drug Discovery: Evaluation of an Inhibitor Activity on Spermidine Synthase of Trypanosoma cruzi
Although
NMR in fragment-based drug discovery is utilized almost
exclusively to evaluate physical binding between molecules, it should
be also a powerful tool for biochemical assay, evaluating inhibitory
effect of compounds on enzymatic activity. Time-dependent spectral
change in real-time monitoring or inhibitor concentration-dependent
spectral change after constant-time reaction was processed by factor
analysis, by which reaction rate or IC<sub>50</sub> value was obtained.
Applications to spermidine synthase of Trypanosoma
cruzi, which causes Chagas disease, are described
4‑Hydroxypyridazin-3(2<i>H</i>)‑one Derivatives as Novel d‑Amino Acid Oxidase Inhibitors
d-Amino acid oxidase (DAAO) catalyzes the oxidation of d-amino acids including d-serine, a coagonist of the <i>N</i>-methyl-d-aspartate receptor. We identified a
series of 4-hydroxypyridazin-3Â(2<i>H</i>)-one derivatives
as novel DAAO inhibitors with high potency and substantial cell permeability
using fragment-based drug design. Comparisons of complex structures
deposited in the Protein Data Bank as well as those determined with
in-house fragment hits revealed that a hydrophobic subpocket was formed
perpendicular to the flavin ring by flipping Tyr224 in a ligand-dependent
manner. We investigated the ability of the initial fragment hit, 3-hydroxy-pyridine-2Â(1<i>H</i>)-one, to fill this subpocket with the aid of complex structure
information. 3-Hydroxy-5-(2-phenylethyl)Âpyridine-2Â(1<i>H</i>)-one exhibited the predicted binding mode and demonstrated high
inhibitory activity for human DAAO in enzyme- and cell-based assays.
We further designed and synthesized 4-hydroxypyridazin-3Â(2<i>H</i>)-one derivatives, which are equivalent to the 3-hydroxy-pyridine-2Â(1<i>H</i>)-one series but lack cell toxicity. 6-[2-(3,5-Difluorophenyl)Âethyl]-4-hydroxypyridazin-3Â(2<i>H</i>)-one was found to be effective against MK-801-induced
cognitive deficit in the Y-maze
NMR Biochemical Assay for Oxidosqualene Cyclase: Evaluation of Inhibitor Activities on <i>Trypanosoma cruzi</i> and Human Enzymes
Oxidosqualene
cyclase (OSC), a membrane-associated protein, is
a key enzyme of sterol biosynthesis. Here we report a novel assay
for OSC, involving reaction in aqueous solution, NMR quantification
in organic solvent, and factor analysis of spectra. We evaluated one
known and three novel inhibitors on OSC of <i>Trypanosoma cruzi</i>, a parasite causative of Chagas disease, and compared their effects
on human OSC for selectivity. Among them, one novel inhibitor showed
a significant parasiticidal activity
NMR Biochemical Assay for Oxidosqualene Cyclase: Evaluation of Inhibitor Activities on <i>Trypanosoma cruzi</i> and Human Enzymes
Oxidosqualene
cyclase (OSC), a membrane-associated protein, is
a key enzyme of sterol biosynthesis. Here we report a novel assay
for OSC, involving reaction in aqueous solution, NMR quantification
in organic solvent, and factor analysis of spectra. We evaluated one
known and three novel inhibitors on OSC of <i>Trypanosoma cruzi</i>, a parasite causative of Chagas disease, and compared their effects
on human OSC for selectivity. Among them, one novel inhibitor showed
a significant parasiticidal activity