Canagliflozin (JNJ 28431754AAA) is a selective SGLT2 inhibitor that inhibits CHO cells expressing mSGLT2, rSGLT2, and hSGLT2 (IC50=2/3.7/4.4 nM). Canagliflozin can be used for the treatment of type II diabetes mellitus (T2DM).

SGLT2 (rat):3.7 nM, SGLT2 (mouse):2 nM, SGLT2 (human):4.4 nM

方法: CHOK-hSGLT1 和 CHOK-hSGLT2 细胞用 Canagliflozin (0.01-10000 nM) 孵育 10 min，检测细胞对钠依赖性葡萄糖的摄取。
结果: Canagliflozin 抑制 CHO-hSGLT1 和 CHOK-hSGLT2 细胞中 14C-AMG 的摄取，IC50 分别为 684±159 nM 和 4.4±1.2 nM。[1]
方法: 人脐静脉内皮细胞 HUVECs 用 Canagliflozin (1-50 µM) 处理 24 h，测量 DNA 合成来监测细胞增殖。
结果: 用 Canagliflozin 处理 HUVECs 导致 DNA 合成的浓度依赖性抑制。在服用 Canagliflozin 的患者血浆中达到的浓度 (5-10 µM) 下，观察到 Canagliflozin 阻断 DNA 合成的能力。此外，更高浓度的 Canagliflozin (50 µM) 几乎消除了 DNA 合成。[2]

方法: 为研究治疗 2 型糖尿病 (T2DM) 的效果，将 Canagliflozin (0.1-30 mg/kg，0.5% hydroxypropyl methylcellulose) 灌胃给药给四种动物模型。动物模型包括：(1) 喂食高脂肪饮食的雄性 C57BL/6J 小鼠；(2) 雄性 C57BL/ksj-db/db 高血糖小鼠；(3) 雄性 Zucker 脂肪 (ZF) 肥胖、胰岛素抵抗大鼠；(4) 雄性 ZDF 肥胖、高血糖大鼠。
结果: 在 T2DM 啮齿动物模型中，Canagliflozin 降低了 RTG 并增加了 UGE，改善了血糖控制和 β 细胞功能，并减少了肥胖啮齿动物模型中的体重增加。[1]

Cells from the rat skeletal muscle cell line, L6, is used to test the effect of Canagliflozin on glucose transporter 1 (GLUT1) activity. Cells are maintained in Dulbecco's modified Eagle's medium containing 5.6 mM glucose supplemented with 10% fetal bovine serum, are seeded in 24-well plates at a density of 3 × 105 cells/well and cultured for 24 hours in an atmosphere of 5% CO2 at 37 °C. Cells are rinsed twice with Kreb's ringer phosphate HEPES buffer (pH 7.4, 150 mM NaCl, 5 mM KCl, 1.25 mM MgSO4, 1.25 mM CaCl2, 2.9 mM Na2HPO4, 10 mM HEPES) and are pre-incubated with the solutions of Canagliflozin (250 μL, 10 μM) for 5 minutes at room temperature. The transport reaction is initiated by adding 50 μL of 4.5 mM 2-DG (a substrate for GLUTs)/3H-2-DG (0.625 μCi) followed by incubation for 15 minutes at room temperature. The 2-DG uptake is halted by aspiration of the incubation mixture. Cells are immediately washed 3 times with ice-cold PBS. Samples are extracted with 0.3 N NaOH, and radioactivity is determined by liquid scintillation.(Only for Reference)