MBX-2982 is a selective, orally-available GPR119 agonist, used for the treatment of diabetes.

在进行了“长期孵育/洗脱”实验并预先处理了MBX-2982 (1 μM) 的细胞中，即便进行了彻底的洗涤以去除过量的激动剂，通过IBMX的包含测得的cAMP积累显著高于对照细胞（P<0.01；ANOVA；n=3-6）。AR-231,453在类似的浓度范围内产生持续反应，与急性刺激观察到的反应相似（小幅上升1.82倍），相应的pEC50分别为8.67±0.11和8.93±0.17。此外，对于MBX-2982，在持续和急性刺激下观察到较大但不那么严重的浓度反应偏移（57.54倍），相应的pEC50分别为7.03±0.13和8.79±0.12[1]。

为探究在GLUTag和主要肠道细胞中的观察结果是否具有生理相关性，C57BL/6小鼠以10 mg/kg的剂量接受GPR119激动剂MBX-2982的处理。注意，本实验中未共同给予DPP-IV抑制剂以考察GPR119的直接效应，但在采集血样时使用了DPP-IV抑制剂以保留血中活性GLP-1。未给予葡萄糖负荷的情况下，接受MBX-2982处理的小鼠的血浆GLP-1水平升高，表明GPR119介导的GLP-1分泌并不依赖于葡萄糖[2]。

HEK-GPR119 cells are transfected with GloSensor 22F plasmid and used for dynamic cAMP measurements 24-30 h later. Cell suspensions are made by dislodging the cells using PBS wash and Accutase treatment followed by resuspension in culture media. Cells are then washed twice by pelleting through centrifugation (300 g, 5 min) and resuspension in assay buffer (Hank's Balanced Salt Solution supplemented with 20 mM HEPES and 0.01% fatty acid free BSA, pH 7.4). Cells are then counted and diluted to 600,000 cells/mL in buffer, before GloSensor cAMP reagent is added (2% v/v) and equilibrated with the cells for 2 h at 20°C with periodic mixing. 50 μl/well of cells are added to white-bottomed 384 well plates (30,000 cells/well) in triplicate and baseline luminescence is measuring using an Envision plate-reader. 5 μL of MBX-2982 (serially diluted in DMSO and then diluted 1:100 in assay buffer to obtain ×10 concentrated solution) is manually added to the assay wells to achieve the stated final concentration. Plates are incubated at 20°C with luminescence read at regular intervals to detect dynamic cAMP changes over time within the same wells. cAMP responses at each time-point are expressed as fold over control (vehicle-treated cells)[1].

MBX-2982 is serially diluted in DMSO and then diluted 1:100 in assay buffer to obtain ×10 concentrated solutionis[1]. HEK-GPR119 cells are grown to confluency in flasks, and cell suspensions are made by dislodging cells using PBS wash and accutase treatment followed by resuspension in culture media. Cells are then washed twice by pelleting through centrifugation (227 g, 7 min, 20°C) and resuspension in warm assay buffer (Hank's Balanced Salt Solution supplemented with 20 mM HEPES and 0.01% fatty acid free BSA, pH 7.4), with a 5 min incubation at 37°C after the second wash. Cells are then counted and diluted to 200,000 cells/mL in warm assay buffer[1].