title: Synthesis and structure-activity relationship study of chemical probes as hypoxia induced factor-1α/malate dehydrogenase 2 inhibitors.
abstract: A structure-activity relationship study of hypoxia inducible factor-1α inhibitor 3-aminobenzoic acid-based chemical probes, which were previously identified to bind to mitochondrial malate dehydrogenase 2, was performed to provide a better understanding of the pharmacological effects of LW6 and its relation to hypoxia inducible factor-1α (HIF-1α) and malate dehydrogenase 2 (MDH2). A variety of multifunctional probes including the benzophenone or the trifluoromethyl diazirine for photoaffinity labeling and click reaction were prepared and evaluated for their biological activity using a cell-based HRE-luciferase assay as well as a MDH2 assay in human colorectal cancer HCT116 cells. Among them, the diazirine probe 4a showed strong inhibitory activity against both HIF-1α and MDH2. Significantly, the inhibitory effect of the probes on HIF-1α activity was consistent with that of the MDH2 enzyme assay, which was further confirmed by the effect on in vitro binding activity to recombinant human MDH2, oxygen consumption, ATP production, and AMP activated protein kinase (AMPK) activation. Competitive binding modes of LW6 and probe 4a to MDH2 were also demonstrated.
authors.affiliation: BK21Plus R-FIND Team, College of Pharmacy, Dongguk University-Seoul , Koyang, 410-820, Korea.
authors.forename: Ravi
authors.initials: R
authors.lastname: Naik
authors.forename: Misun
authors.initials: M
authors.lastname: Won
authors.forename: Hyun Seung
authors.initials: HS
authors.lastname: Ban
authors.forename: Deepak
authors.initials: D
authors.lastname: Bhattarai
authors.forename: Xuezhen
authors.initials: X
authors.lastname: Xu
authors.forename: Yumi
authors.initials: Y
authors.lastname: Eo
authors.forename: Ye Seul
authors.initials: YS
authors.lastname: Hong
authors.forename: Sarbjit
authors.initials: S
authors.lastname: Singh
authors.forename: Yongseok
authors.initials: Y
authors.lastname: Choi
authors.forename: Hee-Chul
authors.initials: HC
authors.lastname: Ahn
authors.forename: Kyeong
authors.initials: K
authors.lastname: Lee
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D001577:Benzophenones / Q000737:chemistry; D001667:Binding, Competitive; D045744:Cell Line, Tumor; D002626:Chemistry, Pharmaceutical / Q000379:methods; D002846:Chromatography, Affinity / Q000379:methods; D015195:Drug Design; D055808:Drug Discovery; D045325:HCT116 Cells; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000037:antagonists & inhibitors*; D020128:Inhibitory Concentration 50; D007700:Kinetics; D010101:Oxygen Consumption; D051736:Proto-Oncogene Proteins c-mdm2 / Q000037:antagonists & inhibitors*; D011994:Recombinant Proteins / Q000737:chemistry; D013329:Structure-Activity Relationship; D062368:meta-Aminobenzoates / Q000737:chemistry
chemical_list: D001577:Benzophenones; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D011994:Recombinant Proteins; D062368:meta-Aminobenzoates; C047723:benzophenone; D000255:Adenosine Triphosphate; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2; C044855:3-aminobenzoic acid