Utilization of an in vivo reporter for high throughput identification of branched small molecule regulators of hypoxic adaptation

Chem Biol. 2010 Apr 23;17(4):380-91. doi: 10.1016/j.chembiol.2010.03.008.

Abstract

Small molecules inhibiting hypoxia inducible factor (HIF) prolyl hydroxylases (PHDs) are the focus of drug development efforts directed toward the treatment of ischemia and metabolic imbalance. A cell-based reporter produced by fusing HIF-1 alpha oxygen degradable domain (ODD) to luciferase was shown to work as a capture assay monitoring stability of the overexpressed luciferase-labeled HIF PHD substrate under conditions more physiological than in vitro test tubes. High throughput screening identified novel catechol and oxyquinoline pharmacophores with a "branching motif" immediately adjacent to a Fe-binding motif that fits selectively into the HIF PHD active site in in silico models. In accord with their structure-activity relationship in the primary screen, the best "hits" stabilize HIF1 alpha, upregulate known HIF target genes in a human neuronal line, and exert neuroprotective effects in established model of oxidative stress in cortical neurons.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catechols / chemistry
  • Catechols / pharmacology*
  • Cell Line, Tumor
  • Dioxygenases / antagonists & inhibitors*
  • Dioxygenases / chemistry
  • Dioxygenases / metabolism
  • High-Throughput Screening Assays / methods*
  • Humans
  • Hydrazines / chemistry
  • Hydrazines / pharmacology
  • Hypoxia / drug therapy*
  • Hypoxia-Inducible Factor 1 / metabolism
  • Hypoxia-Inducible Factor-Proline Dioxygenases
  • Models, Molecular
  • Neuroprotective Agents / chemistry
  • Neuroprotective Agents / pharmacology
  • Nuclear Proteins / antagonists & inhibitors*
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / metabolism
  • Oxidative Stress / drug effects
  • Oxyquinoline / chemistry
  • Oxyquinoline / pharmacology*
  • Procollagen-Proline Dioxygenase
  • Structure-Activity Relationship
  • Thiadiazoles / chemistry
  • Thiadiazoles / pharmacology

Substances

  • Catechols
  • Hydrazines
  • Hypoxia-Inducible Factor 1
  • Neuroprotective Agents
  • Nuclear Proteins
  • Thiadiazoles
  • Oxyquinoline
  • Dioxygenases
  • Procollagen-Proline Dioxygenase
  • EGLN2 protein, human
  • Hypoxia-Inducible Factor-Proline Dioxygenases
  • catechol