Utilization of an in vivo reporter for high throughput identification of branched small molecule regulators of hypoxic adaptation.
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.
Smirnova
Natalya A
NA
Burke Medical Research Institute, Department of Neurology and Neuroscience, Weill Medical College of Cornell University, 785 Mamaroneck Ave, White Plains, NY 10605, USA.
Rakhman
Ilay
I
Moroz
Natalia
N
Basso
Manuela
M
Payappilly
Jimmy
J
Kazakov
Sergey
S
Hernandez-Guzman
Francisco
F
Gaisina
Irina N
IN
Kozikowski
Alan P
AP
Ratan
Rajiv R
RR
Gazaryan
Irina G
IG
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
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