RESUMO
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.
Assuntos
Descoberta de Drogas , Corantes Fluorescentes/química , Glucose/análogos & derivados , Hipoglicemiantes/farmacologia , Moduladores de Transporte de Membrana/farmacologia , Naftalimidas/química , Inibidores do Transportador 2 de Sódio-Glicose , Absorção Fisiológica/efeitos dos fármacos , Animais , Células CHO , Radioisótopos de Carbono , Química Click , Células Clonais , Cricetulus , Corantes Fluorescentes/análise , Glucose/metabolismo , Ensaios de Triagem em Larga Escala , Humanos , Cinética , Metilglucosídeos/metabolismo , Naftalimidas/análise , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Bibliotecas de Moléculas Pequenas , Transportador 2 de Glucose-Sódio/genética , Transportador 2 de Glucose-Sódio/metabolismoAssuntos
Mobilização de Células-Tronco Hematopoéticas , Receptores CXCR4/antagonistas & inibidores , Animais , Benzilaminas , Ciclamos , Avaliação Pré-Clínica de Medicamentos , Células-Tronco Hematopoéticas/citologia , Compostos Heterocíclicos/química , Compostos Heterocíclicos/farmacologia , Injeções Subcutâneas , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Poliaminas/química , Pirimidinas/química , Quinazolinas/síntese química , Quinazolinas/química , Quinazolinas/farmacologia , Receptores CXCR4/metabolismo , Medicina RegenerativaRESUMO
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.
Assuntos
Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Tipo 2/tratamento farmacológico , Glucosídeos/síntese química , Hiperglicemia/tratamento farmacológico , Hipoglicemiantes/síntese química , Indóis/síntese química , Inibidores do Transportador 2 de Sódio-Glicose , Xilose/análogos & derivados , Animais , Células CHO , Cricetinae , Cricetulus , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/urina , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/urina , Glucose/metabolismo , Glucosídeos/farmacocinética , Glucosídeos/farmacologia , Humanos , Hipoglicemiantes/farmacocinética , Hipoglicemiantes/farmacologia , Indóis/farmacocinética , Indóis/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos ICR , Ratos , Ratos Sprague-Dawley , Relação Estrutura-Atividade , Xilose/síntese química , Xilose/farmacocinética , Xilose/farmacologiaRESUMO
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.