Canagliflozin is a selective SGLT2 inhibitor with IC₅₀ of 2 nM, 3.7 nM, and 4.4 nM for mSGLT2, rSGLT2, and hSGLT2 in CHOK cells, respectively.

IC50: 2/3.7/4.4 nM (m/r/hSGLT2, in CHOK cells)^[1]

Canagliflozin is a sodium glucose co-transporter (SGLT) 2 inhibitor. In a concentration-dependent fashion, Canagliflozin inhibits Na⁺-dependent ¹⁴C-AMG uptake in CHO-hSGLT2 cells, with an IC₅₀ of 4.4±1.2 nM. Similar IC₅₀ values are obtained in CHO-rSGLT2 and CHO-mSGLT2 cells (IC₅₀=3.7 and 2.0 nM for rat and mouse SGLT2, respectively). Canagliflozin inhibits ¹⁴C-AMG uptake in CHO-hSGLT1 and mSGLT1 cells with IC₅₀ of 684±159 nM and >1,000 nM, respectively. At 10 µM, Canagliflozin inhibits the facilitative (non-Na⁺-linked) GLUT-mediated 3H-2-DG uptake in L6 myoblasts by less than 50%^[1].

Canagliflozin treatment (1 mg/kg) notably lowers renal threshold for glucose excretion (RT_G) in Zucker diabetic fatty (ZDF) rats to 94±10 mg/dL. In the second study, an insulin infusion is given to lower blood glucose (BG) to approximately 25 mg/dL, and then the graded glucose infusion (GGI) is given to slowly raise BG to approximately 300 mg/dL. In ZDF rats treated with Canagliflozin (1 mg/kg), the relationship between BG and urinary glucose excretion (UGE) remains well-described by a threshold relationship with negligible UGE occurring when BG<90 mg/dl="" and="" uge="" increases="" in="" proportion="" to="" bg="" at="" higher="" bg="" levels.="" in="" db/db="" mice,="" single="" doses="" of="" canagliflozin="" dose-dependently="" reduces="" non-fasting="" bg="" concentrations.="" the="" onset="" of="" the="" bg-lowering="" effect="" after="" a="" single="" dose="" is="" rapid,="" and="" bg="" levels="" in="" canagliflozin-treated="" mice="" (at="" 1="" and="" 10="" mg/kg="" doses)="" are="" significantly="" different="" from="" those="" of="" vehicle-treated="" mice="" at="" 1="" hour="" after="" treatment="" (22%="" and="" 36%="" reduction="" of="" bg="" levels="" compared="" with="" that="" in="" vehicle-treated="" mice,="">^[1].

• [1]. Liang Y, et al. Effect of canagliflozin on renal threshold for glucose, glycemia, and body weight in normal and diabetic animal models. PLoS One. 2012;7(2):e30555.

  [2]. Kobuchi S, et al. A validated LC-MS/MS method for the determination of canagliflozin, a sodium-glucose co-transporter 2 (SGLT-2) inhibitor, in a lower volume of rat plasma: application to pharmacokinetic studies in rats. Biomed Chromatogr. 2016 Oct;30(10):1549-55.

Canagliflozin is dissolved in DMSO and stored, and then diluted with appropriate media (DMSO 0.1%) before use^[1].

Sodium-dependent glucose uptake in Chinese Hamster Ovary (CHO) cells expressing SGLT1 and SGLT2 co-transporters parental CHO-K (CHOK) cells (commonly used mammalian cells for gene overexpression studies) expressing human or mouse SGLT1 and SGLT2 are utilized in this study. Cells are seeded into 96-well plates. Cells are then washed one time with 0.15 mL assay buffer (137 mM NaCl, 5 mM KCl, 1 mM CaCl₂, 1 mM MgCl₂, 50 mM HEPES, pH 7.4) at 37°C. After the assay buffer is removed, 50 µL of fresh assay buffer with 5 µL of Canagliflozin (0.3-300 nM) is added, followed by 10 minutes of incubation. Then, 5 µL of 6 mM alpha-methyl-D-glucopyranoside (AMG, a selective SGLT1/2 substrate)/¹⁴C-AMG (0.07 µCi) is added to the cells and incubated at 37°C for 2 hours. Next, the cells are washed 3 times with 0.15 mL ice-cold phosphate-buffered saline (PBS). After the final wash is aspirated, 50 µL of microscint 20 is added. The plate is counted by TopCount^[1]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Canagliflozin is formulated in 0.5% hydroxypropyl methylcellulose^[1].

Mice and Rat^[1]
Four rodent models are used in these experiments: (1) male C57BL/6J mice fed with a high-fat diet (D-12492 with 60 kcal% fat) (diet-induced obese, insulin resistantmice [DIO]); (2) male C57BL/ksj-db/db hyperglycemic mice; (3) male Zucker fatty (ZF) obese, insulin resistant rats; and (4) male ZDF obese, hyperglycemic rats. To examine the effect of Canagliflozin on hyperglycemia, single doses of Canagliflozin (0.1, 1, and 10 mg/kg) are administered to overnight-fasted db/db mice. BG levels are monitored at 0, 0.5, 1, 3, 6, and 24 hours after dosing. Canagliflozin is also administered to ZDF rats at varying doses (3-30 mg/kg) for 4 weeks to evaluate its effect on BG control and pancreatic beta-cell function. BG levels are monitored weekly, and HbA1c, plasma glucose, and insulin levels are determined at the end of the 4-week treatment. An oral glucose tolerance test (OGTT) (2 mg/kg of body weight, given by gavage) is conducted in ZDF rats after 4 weeks of treatment. Blood is sampled at 0, 30, 60, and 120 minutes after glucose challenge from the tail vein for measurement of BG levels using a glucometer and plasma insulin using ELISA method. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Liang Y, et al. Effect of canagliflozin on renal threshold for glucose, glycemia, and body weight in normal and diabetic animal models. PLoS One. 2012;7(2):e30555.

  [2]. Kobuchi S, et al. A validated LC-MS/MS method for the determination of canagliflozin, a sodium-glucose co-transporter 2 (SGLT-2) inhibitor, in a lower volume of rat plasma: application to pharmacokinetic studies in rats. Biomed Chromatogr. 2016 Oct;30(10):1549-55.

444.52

C₂₄H₂₅FO₅S

842133-18-0

Room temperature in continental US; may vary elsewhere

10 mM in DMSO

Canagliflozin is prepared in 1% carboxymethyl-cellulose sodium in distilled water^[2].

* "<1 mg/ml"="" means="" slightly="" soluble="" or="" insoluble.="" "≥"="" means="" soluble,="" but="" saturation="">

• [1]. Liang Y, et al. Effect of canagliflozin on renal threshold for glucose, glycemia, and body weight in normal and diabetic animal models. PLoS One. 2012;7(2):e30555.

  [2]. Kobuchi S, et al. A validated LC-MS/MS method for the determination of canagliflozin, a sodium-glucose co-transporter 2 (SGLT-2) inhibitor, in a lower volume of rat plasma: application to pharmacokinetic studies in rats. Biomed Chromatogr. 2016 Oct;30(10):1549-55.