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.
Subject(s)
Drug Discovery , Fluorescent Dyes/chemistry , Glucose/analogs & derivatives , Hypoglycemic Agents/pharmacology , Membrane Transport Modulators/pharmacology , Naphthalimides/chemistry , Sodium-Glucose Transporter 2 Inhibitors , Absorption, Physiological/drug effects , Animals , CHO Cells , Carbon Radioisotopes , Click Chemistry , Clone Cells , Cricetulus , Fluorescent Dyes/analysis , Glucose/metabolism , High-Throughput Screening Assays , Humans , Kinetics , Methylglucosides/metabolism , Naphthalimides/analysis , Recombinant Proteins/chemistry , Recombinant Proteins/metabolism , Small Molecule Libraries , Sodium-Glucose Transporter 2/genetics , Sodium-Glucose Transporter 2/metabolismSubject(s)
Hematopoietic Stem Cell Mobilization , Receptors, CXCR4/antagonists & inhibitors , Animals , Benzylamines , Cyclams , Drug Evaluation, Preclinical , Hematopoietic Stem Cells/cytology , Heterocyclic Compounds/chemistry , Heterocyclic Compounds/pharmacology , Injections, Subcutaneous , Male , Mice , Mice, Inbred C57BL , Polyamines/chemistry , Pyrimidines/chemistry , Quinazolines/chemical synthesis , Quinazolines/chemistry , Quinazolines/pharmacology , Receptors, CXCR4/metabolism , Regenerative MedicineABSTRACT
A novel series of N-linked ß-D-xylosides were synthesized and evaluated for inhibitory activity against sodium-dependent glucose cotransporter 2 (SGLT2) in a cell-based assay. Of these, the 4-chloro-3-(4-cyclopropylbenzyl)-1-(ß-D-xylopyranosyl)-1H-indole 19m was found to be the most potent inhibitor, with an EC(50) value similar to that of the natural SGLT2 inhibitor phlorizin. Further studies in Sprague-Dawley (SD) rats indicated that 19m significantly increased urine glucose excretion in a dose-dependent manner with oral administration. The antihyperglycemic effect of 19m was also observed in streptozotocin (STZ) induced diabetic SD rats. These results described here are a good starting point for further investigations into N-glycoside SGLT2 inhibitors.
Subject(s)
Diabetes Mellitus, Experimental/drug therapy , Diabetes Mellitus, Type 2/drug therapy , Glucosides/chemical synthesis , Hyperglycemia/drug therapy , Hypoglycemic Agents/chemical synthesis , Indoles/chemical synthesis , Sodium-Glucose Transporter 2 Inhibitors , Xylose/analogs & derivatives , Animals , CHO Cells , Cricetinae , Cricetulus , Diabetes Mellitus, Experimental/blood , Diabetes Mellitus, Experimental/urine , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/urine , Glucose/metabolism , Glucosides/pharmacokinetics , Glucosides/pharmacology , Humans , Hypoglycemic Agents/pharmacokinetics , Hypoglycemic Agents/pharmacology , Indoles/pharmacokinetics , Indoles/pharmacology , Male , Mice , Mice, Inbred ICR , Rats , Rats, Sprague-Dawley , Structure-Activity Relationship , Xylose/chemical synthesis , Xylose/pharmacokinetics , Xylose/pharmacologyABSTRACT
A ligand-based virtual screening strategy (a combination of pharmacophore model generation, shape-based scoring, and structure clustering analysis) was developed to discover novel SGLT2 inhibitors. The best pharmacophore model, generated from eight glycoside inhibitors, was utilized to virtually screen three chemical databases that led to the identification of three non-glycoside SGLT2 inhibitors. This is the first report of the generation of a pharmacophore model from glycosides that has then been used to discover novel non-glycosides hits.
