title: Utilization of an in vivo reporter for high throughput identification of branched small molecule regulators of hypoxic adaptation.
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.
authors.affiliation: Burke Medical Research Institute, Department of Neurology and Neuroscience, Weill Medical College of Cornell University, 785 Mamaroneck Ave, White Plains, NY 10605, USA.
authors.forename: Natalya A
authors.initials: NA
authors.lastname: Smirnova
authors.forename: Ilay
authors.initials: I
authors.lastname: Rakhman
authors.forename: Natalia
authors.initials: N
authors.lastname: Moroz
authors.forename: Manuela
authors.initials: M
authors.lastname: Basso
authors.forename: Jimmy
authors.initials: J
authors.lastname: Payappilly
authors.forename: Sergey
authors.initials: S
authors.lastname: Kazakov
authors.forename: Francisco
authors.initials: F
authors.lastname: Hernandez-Guzman
authors.forename: Irina N
authors.initials: IN
authors.lastname: Gaisina
authors.forename: Alan P
authors.initials: AP
authors.lastname: Kozikowski
authors.forename: Rajiv R
authors.initials: RR
authors.lastname: Ratan
authors.forename: Irina G
authors.initials: IG
authors.lastname: Gazaryan
mesh_terms: D002396:Catechols / Q000737:chemistry / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049308:Dioxygenases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D006834:Hydrazines / Q000737:chemistry / Q000494:pharmacology; D000860:Hypoxia / Q000188:drug therapy*; D051793:Hypoxia-Inducible Factor 1 / Q000378:metabolism; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases; D008958:Models, Molecular; D018696:Neuroprotective Agents / Q000737:chemistry / Q000494:pharmacology; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D015125:Oxyquinoline / Q000737:chemistry / Q000494:pharmacology*; D011393:Procollagen-Proline Dioxygenase; D013329:Structure-Activity Relationship; D013830:Thiadiazoles / Q000737:chemistry / Q000494:pharmacology
chemical_list: D002396:Catechols; D006834:Hydrazines; D051793:Hypoxia-Inducible Factor 1; D018696:Neuroprotective Agents; D009687:Nuclear Proteins; D013830:Thiadiazoles; D015125:Oxyquinoline; D049308:Dioxygenases; D011393:Procollagen-Proline Dioxygenase; C447780:EGLN2 protein, human; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases; C034221:catechol