title: Development and application of a fluorescent glucose uptake assay for the high-throughput screening of non-glycoside SGLT2 inhibitors.
abstract: Sodium-dependent glucose co-transporter 2 (SGLT2) inhibitors are of current interest as a treatment for type 2 diabetes. Efforts have been made to discover phlorizin-related glycosides with good SGLT2 inhibitory activity. To increase structural diversity and better understand the role of non-glycoside SGLT2 inhibitors on glycemic control, we initiated a research program to identify non-glycoside hits from high-throughput screening. Here, we report the development of a novel, fluorogenic probe-based glucose uptake system based on a Cu(I)-catalyzed [3+2] cycloaddition. The safer processes and cheaper substances made the developed assay our first priority for large-scale primary screening as compared to the well-known [(14)C]-labeled α-methyl-D-glucopyranoside ([(14)C]-AMG) radioactive assay. This effort culminated in the identification of a benzimidazole, non-glycoside SGLT2 hit with an EC50 value of 0.62 μM by high-throughput screening of 41,000 compounds.
authors.affiliation: Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, 35 Keyan Road, Zhunan Town, Miaoli County 35053, Taiwan, ROC.
authors.forename: Szu-Huei
authors.initials: SH
authors.lastname: Wu
authors.affiliation: Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, 35 Keyan Road, Zhunan Town, Miaoli County 35053, Taiwan, ROC.
authors.forename: Chun-Hsu
authors.initials: CH
authors.lastname: Yao
authors.affiliation: Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, 35 Keyan Road, Zhunan Town, Miaoli County 35053, Taiwan, ROC.
authors.forename: Chieh-Jui
authors.initials: CJ
authors.lastname: Hsieh
authors.affiliation: Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, 35 Keyan Road, Zhunan Town, Miaoli County 35053, Taiwan, ROC.
authors.forename: Yu-Wei
authors.initials: YW
authors.lastname: Liu
authors.affiliation: Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, 35 Keyan Road, Zhunan Town, Miaoli County 35053, Taiwan, ROC.
authors.forename: Yu-Sheng
authors.initials: YS
authors.lastname: Chao
authors.affiliation: Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, 35 Keyan Road, Zhunan Town, Miaoli County 35053, Taiwan, ROC. Electronic address: jssong@nhri.org.tw.
authors.forename: Jen-Shin
authors.initials: JS
authors.lastname: Song
authors.affiliation: Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, 35 Keyan Road, Zhunan Town, Miaoli County 35053, Taiwan, ROC. Electronic address: jinqchyi@nhri.org.tw.
authors.forename: Jinq-Chyi
authors.initials: JC
authors.lastname: Lee
mesh_terms: D065567:Absorption, Physiological / Q000187:drug effects; D000818:Animals; D016466:CHO Cells; D002250:Carbon Radioisotopes; D057930:Click Chemistry; D002999:Clone Cells; D003412:Cricetulus; D055808:Drug Discovery; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry*; D005947:Glucose / Q000031:analogs & derivatives* / Q000378:metabolism; D057166:High-Throughput Screening Assays; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D007700:Kinetics; D049990:Membrane Transport Modulators / Q000494:pharmacology*; D008757:Methylglucosides / Q000378:metabolism; D053644:Naphthalimides / Q000032:analysis / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D054852:Small Molecule Libraries; D051297:Sodium-Glucose Transporter 2 / Q000235:genetics / Q000378:metabolism; D000077203:Sodium-Glucose Transporter 2 Inhibitors
chemical_list: C000604725:4-ethynyl-N-ethyl-1,8-naphthalimide; D002250:Carbon Radioisotopes; D005456:Fluorescent Dyes; D007004:Hypoglycemic Agents; D049990:Membrane Transport Modulators; D008757:Methylglucosides; D053644:Naphthalimides; D011994:Recombinant Proteins; C089180:SLC5A2 protein, human; D054852:Small Molecule Libraries; D051297:Sodium-Glucose Transporter 2; D000077203:Sodium-Glucose Transporter 2 Inhibitors; D005947:Glucose
keywords: Click chemistry; High-throughput screening; Non-glycoside; Sodium-dependent glucose co-transporter; Type 2 diabetes mellitus