title: Molecular and functional evaluation of a novel HIF inhibitor, benzopyranyl 1,2,3-triazole compound.
abstract: Hypoxia occurs in a variety of pathological events, including the formation of solid tumors. Hypoxia-inducible factor (HIF)-1α is stabilized under hypoxic conditions and is a key molecule in tumor growth and angiogenesis. Seeking to develop novel cancer therapeutics, we investigated small molecules from our in-house chemical libraries to target HIF-1α. We employed a dual-luciferase assay that uses a luciferase (Luc) reporter vector harboring five copies of hypoxia-responsive element (HRE) in the promoter. Under hypoxic conditions that increased Luc reporter activity by four-fold, we screened 144 different compounds, nine of which showed 30-50% inhibition of hypoxia-induced Luc reporter activity. Among these, "Compound 12, a benzopyranyl 1,2,3-triazole" was the most efficient at inhibiting the expression of HIF-1α under hypoxic conditions, reducing its expression by 80%. Under hypoxic conditions, the half maximal IC50 of the compound was 24 nM in HEK-293 human embryonic kidney cells, and 2 nM in A549 human lung carcinoma cells. Under hypoxic conditions, Compound 12 increased hydroxylated HIF-1α levels and HIF-1α ubiquitination, and also dose-dependently decreased HIF-1α target gene expression as well as vascular endothelial growth factor (VEGF) secretion. Furthermore, this compound inhibited VEGF-induced in vitro angiogenesis in human umbilical vein endothelial cells (HUVECs), and in vivo, it inhibited chick chorioallantoic membrane angiogenesis. In allogaft assays, cotreatment with Compound 12 and gefitinib significantly inhibited tumor growth and angiogenesis. Compound 12 can be a novel inhibitor of HIF-1α by accelerating its degradation, and shows much potential as an anti-cancer agent through its ability to suppress tumor growth and angiogenesis.
authors.affiliation: BK21 Plus KNU Multi-Omics based Creative Drug Research Team, National Basic Research Laboratory of Vascular Homeostasis Regulation, Kyungpook National University, Buk-gu, 702-701, Daegu, Republic of Korea.
authors.forename: Kyunghye
authors.initials: K
authors.lastname: Park
authors.affiliation: BK21 Plus KNU Multi-Omics based Creative Drug Research Team, National Basic Research Laboratory of Vascular Homeostasis Regulation, Kyungpook National University, Buk-gu, 702-701, Daegu, Republic of Korea.
authors.forename: Hye Eun
authors.initials: HE
authors.lastname: Lee
authors.affiliation: BK21 Plus KNU Multi-Omics based Creative Drug Research Team, National Basic Research Laboratory of Vascular Homeostasis Regulation, Kyungpook National University, Buk-gu, 702-701, Daegu, Republic of Korea.
authors.forename: Sun Hee
authors.initials: SH
authors.lastname: Lee
authors.affiliation: College of Pharmacy, Research Institute of Pharmaceutical Sciences, Kyungpook National University, Buk-gu, 702-701, Daegu, Republic of Korea.
authors.forename: Doohyun
authors.initials: D
authors.lastname: Lee
authors.affiliation: College of Pharmacy, Research Institute of Pharmaceutical Sciences, Kyungpook National University, Buk-gu, 702-701, Daegu, Republic of Korea.
authors.forename: Taeho
authors.initials: T
authors.lastname: Lee
authors.affiliation: BK21 Plus KNU Multi-Omics based Creative Drug Research Team, National Basic Research Laboratory of Vascular Homeostasis Regulation, Kyungpook National University, Buk-gu, 702-701, Daegu, Republic of Korea.|College of Pharmacy, Research Institute of Pharmaceutical Sciences, Kyungpook National University, Buk-gu, 702-701, Daegu, Republic of Korea.
authors.forename: You Mie
authors.initials: YM
authors.lastname: Lee
mesh_terms: D000072283:A549 Cells; D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D018827:Carcinoma, Lewis Lung / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000098:blood supply / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D006900:Hydroxylation; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008810:Mice, Inbred C57BL; D018919:Neovascularization, Physiologic / Q000187:drug effects; D055550:Protein Stability; D059748:Proteolysis; D020218:Response Elements; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D014162:Transfection; D014230:Triazoles / Q000494:pharmacology*; D047368:Tumor Burden / Q000187:drug effects; D000072258:Tumor Hypoxia; D059016:Tumor Microenvironment; D054875:Ubiquitination; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D020533:Angiogenesis Inhibitors; C497442:HIF1A protein, human; C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D014230:Triazoles; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A
keywords: 2; 3-triazole; HIF-1α inhibitor; benzopyranyl 1; chemical library