title: Biomolecular Interaction Assays Identified Dual Inhibitors of Glutaminase and Glutamate Dehydrogenase That Disrupt Mitochondrial Function and Prevent Growth of Cancer Cells.
abstract: Glutaminase (KGA/isoenzyme GAC) is an emerging and important drug target for cancer. Traditional methods for assaying glutaminase activity are coupled with several other enzymes. Such coupled assays do not permit the direct and stringent characterization of specific glutaminase inhibitors. Ebselen was identified as a potent 9 nM KGA inhibitor in the KGA/glutamate oxidase (GO)/horse radish peroxidase (HRP) coupled assay but showed very weak activity in inhibiting the growth of glutamine-dependent cancer cells. For rigorous characterization, we developed a direct kinetic binding assay for KGA using bio-layer interferometry (BLI) as the detection method; Ebselen was identified as a GDH inhibitor but not a KGA inhibitor. Furthermore, we designed and synthesized several benzo[d][1,2]selenazol-3(2H)-one dimers which were subjected to SAR analysis by several glutaminolysis specific biochemical and cell based assays. Novel glutamate dehydrogenase (GDH) or dual KGA/GDH inhibitors were discovered from the synthetic compounds; the dual inhibitors completely disrupt mitochondrial function and demonstrate potent anticancer activity with a minimum level of toxicity.
authors.affiliation: School of Pharmacy, Zhejiang University of Technology , Hangzhou, Zhejiang, China 310014.
authors.forename: Min
authors.initials: M
authors.lastname: Zhu
authors.affiliation: School of Pharmacy, Zhejiang University of Technology , Hangzhou, Zhejiang, China 310014.
authors.forename: Jinzhang
authors.initials: J
authors.lastname: Fang
authors.affiliation: School of Pharmacy, Zhejiang University of Technology , Hangzhou, Zhejiang, China 310014.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: School of Pharmacy, Zhejiang University of Technology , Hangzhou, Zhejiang, China 310014.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: School of Pharmacy, Zhejiang University of Technology , Hangzhou, Zhejiang, China 310014.
authors.forename: Jianhui
authors.initials: J
authors.lastname: Xie
authors.affiliation: School of Pharmacy, Zhejiang University of Technology , Hangzhou, Zhejiang, China 310014.
authors.forename: Yan
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Hangzhou Jennifer Biotech. Inc. , Hangzhou, China 31009.
authors.forename: Jennifer Jin
authors.initials: JJ
authors.lastname: Ruan
authors.affiliation: School of Pharmacy, Zhejiang University of Technology , Hangzhou, Zhejiang, China 310014.
authors.forename: Zhao
authors.initials: Z
authors.lastname: Chen
authors.affiliation: School of Pharmacy, Zhejiang University of Technology , Hangzhou, Zhejiang, China 310014.
authors.forename: Wei
authors.initials: W
authors.lastname: Hou
authors.affiliation: School of Pharmacy, Zhejiang University of Technology , Hangzhou, Zhejiang, China 310014.
authors.forename: Gensheng
authors.initials: G
authors.lastname: Yang
authors.affiliation: School of Pharmacy, Zhejiang University of Technology , Hangzhou, Zhejiang, China 310014.
authors.forename: Weike
authors.initials: W
authors.lastname: Su
authors.affiliation: School of Pharmacy, Zhejiang University of Technology , Hangzhou, Zhejiang, China 310014.
authors.forename: Benfang Helen
authors.identifier: 0000-0001-9981-8328
authors.initials: BH
authors.lastname: Ruan
mesh_terms: D000495:Allosteric Site; D001393:Azoles / Q000032:analysis* / Q000378:metabolism / Q000494:pharmacology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D057075:Enzyme Assays; D004791:Enzyme Inhibitors / Q000032:analysis* / Q000378:metabolism / Q000494:pharmacology; D005969:Glutamate Dehydrogenase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D005972:Glutaminase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D054833:Isoindoles; D007700:Kinetics; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D016566:Organoselenium Compounds / Q000032:analysis* / Q000378:metabolism / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D001393:Azoles; D004791:Enzyme Inhibitors; D054833:Isoindoles; D016566:Organoselenium Compounds; D011994:Recombinant Proteins; C042986:ebselen; D005969:Glutamate Dehydrogenase; D005972:Glutaminase