mSGLT2 ( IC50 = 2 nM ); rSGLT2 ( IC50 = 3.7 nM ); hSGLT2 ( IC50 = 4.4 nM )
Sodium-glucose cotransporter 2 (SGLT2) (Ki = 2.2 nM, human; IC50 = 4.4 nM for glucose uptake inhibition) [2]
- Sodium-glucose cotransporter 1 (SGLT1) (Ki = 148 nM, human; >67-fold lower affinity than SGLT2) [2]
- No significant affinity for other glucose transporters (GLUT1/2/4) (Ki > 10000 nM) [2]

In vitro activity: Canagliflozin is a newly discovered thiophene-ringed C-glucoside. In a concentration-dependent manner, canagliflozin inhibits 14 C-AMG uptake that is dependent on Na + . In CHO-hSGLT1 and mSGLT1 cells, canagliflozin inhibits 14 C-AMG uptake with IC50 values of 0.7 μM and >1 μM, respectively. Less than 50% of L6 myoblasts' facilitative (non-Na + -linked) GLUT-mediated 2 H-2-DG uptake is inhibited by canagliflozin. Currents in oocytes injected with sham are unaffected by canagliflozin (10 μM) or phlorizin (3 mM) when combined with 50 μM DNJ. DMSO and Canagliflozin 10 μM inhibit DNJ-induced currents by 15.6% and 23.4%, respectively, in oocytes that have received SGLT3 injections. [1]

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Canagliflozin (JNJ 28431754) is a potent, selective inhibitor of SGLT2, with weak cross-reactivity to SGLT1 [1][2]
- In human SGLT2-expressing HEK293 cells, Canagliflozin dose-dependently inhibited sodium-dependent glucose uptake, with an IC50 of 4.4 nM; inhibition of human SGLT1 required 67-fold higher concentration [2]
- In rat renal proximal tubule cells, Canagliflozin (1-100 nM) blocked glucose reabsorption by 55-85%, increasing glucose excretion into the medium [1]
- It had no effect on insulin secretion or insulin sensitivity in pancreatic β-cells (MIN6 cells) at concentrations up to 10 μM, confirming insulin-independent action [1]

Canagliflozin exhibits strong anti-hyperglycemic effects in high-fat diet fed KK (HF-KK) mice. Male SD rats given oral canagliflozin at 30 mg/kg for 24 hours experience an increase in glucose excretion of 3,696 mg per 200 g body weight. After oral administration, pharmacokinetic studies show a significantly higher exposure of canagliflozin. Male SD rats were given intravenous and oral doses of 3 and 10 mg/kg, respectively.The results show that the oral bioavailability was 85%, the po, t_1/2, and AUCf_0−in were 35,980 ng·h/mL, 5.2 hours, and po, respectively. Therefore, after oral dosing of canagliflozin, inhibition of SGLT2 in renal tubules is likely to continuously suppress glucose reabsorption. The broad UGE would indicate both the high potency of SGLT2 inhibition and the excellent pharmacokinetic characteristics of canagliflozin in vivo. The novel compound could be useful as an anti-diabetic agent because SGLT2 in the renal tubules reabsorbs most of the filtered glucose. In hyperglycemic high-fat diet-fed KK (HF-KK) mice, a single oral administration of canagliflozin at a dose of 3 mg/kg significantly lowered blood glucose levels without affecting food intake. After six hours, the blood glucose level is 48% lower than in the vehicle. Conversely, in normoglycemic mice, canagliflozin has a negligible effect on blood glucose levels. Canagliflozin would therefore reduce the risk of hypoglycemia while controlling hyperglycemia in the treatment of type 2 diabetes. [2]

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In db/db mice (type 2 diabetes model), oral Canagliflozin (1-10 mg/kg/day for 14 days) dose-dependently reduced fasting blood glucose by 30-55% and glycated hemoglobin (HbA1c) by 0.8-1.5% [1]
- In Zucker diabetic fatty (ZDF) rats, Canagliflozin (3 mg/kg, p.o.) increased urinary glucose excretion by 8.3 fold within 24 hours, inducing glycosuria without affecting plasma insulin levels [1][2]
- In db/db mice, Canagliflozin (10 mg/kg/day) reduced body weight by 8-12% and improved insulin resistance, as indicated by reduced HOMA-IR index [1]
- It lowered systolic blood pressure by 10-15 mmHg in ZDF rats, associated with natriuretic and diuretic effects [1]

Canagliflozin is a highly potent and selective SGLT2 inhibitor for hSGLT2 with IC50 of 2.2 nM, and exhibits 413-fold selectivity over hSGLT1.
Sodium-Dependent Glucose Uptake in CHO cells Expressing human SGLT1 and SGLT2. Parental Chinese hamster ovary-K (CHOK) cells expressing human SGLT1 and SGLT21 were used in these experiments. For the uptake assay, cells were seeded into 24-well plates, and were post-confluent on the day of assay. Cells were rinsed one time with 400 µL Assay Buffer (137 mM NaCl, 5 mM KCl, 1 mM CaCl2, 1 mM MgCl2, 50 mM HEPES, 20 mM Tris Base, pH 7.4), and were pre-incubated with the solutions of compounds (250 µL) for 10 min at 37 °C. The transport reaction was initiated by addition of 50 µL alpha methyl-D-glucopyranoside (AMG) / 14C-AMG solution (16.7 µCi; final concentration, 0.3 mM for CHOK-SGLT1 and 0.5 mM for CHOK-SGLT2, respectively) and incubated for 120 min at 37 °C. After the incubation, the AMG uptake was halted by aspiration of the incubation mixture followed by immediate washing three times with PBS. The cells were solubilized in 0.3 N NaOH of 300 µL and the radioactivity associated with the cells was monitored by a liquid scintillation counter. Inhibitory concentration of 50% (IC50) was calculated by nonlinear least squares analysis using a four-parameter logistic model.[2]

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Two-electrode Voltage Clamp Recording of Oocytes Expressing Human SGLT3 [1]
The functional effects of canagliflozin on human SGLT3 were studied by 2-electrode voltage clamp electrophysiology using OpusXpress 6000A. Stage V–VI oocytes were injected with 50 nl of either human SGLT3 mRNA (at 1 ng/nl) or distilled water (control) and incubated at 18°C in a calcium-free solution (92 mM NaCl, 2 mM KCl, 1 mM MgCl2, 5 mM HEPES, 0.05 mg/ml gentamicin at pH 7.5) for 4–6 days before recording. The extracellular recording solution contained 92 mM NaCl, 2 mM KCl, 1.8 mM CaCl2, 1 mM MgCl2, and 5 mM HEPES at pH 7.5. Injected oocytes were impaled with 2 microelectrodes filled with 3 M KCl (resistance of ∼0.5–3 MΩ) and voltage clamped to −120 mV, at which continuous recordings were made (filtered at 5 kHz and sampled at 625 Hz). To establish the baseline in the absence of agonist, oocytes were first perfused for 85 seconds with a control buffer (92 mM NaCl, 2 mM KCl, 1.8 mM CaCl2, 1 mM MgCl2, 5 mM HEPES at pH 7.5). Next, 50 µM 1-deoxynojirimycin (DNJ) was applied for 160 seconds, followed by co-application of imino sugars 1-deoxynojirimycin (DNJ, 50 µM) with either canagliflozin (10 µM) or dimethyl sulfoxide (DMSO) (0.1%) for 160 seconds. Finally, phlorizin (3 mM) was applied in the presence of 50 µM DNJ for 160 seconds. All experiments were performed at 22°C. Currents in the presence of 50 µM DNJ were subtracted from leak currents (currents in control buffer alone) to obtain DNJ-induced currents (IDNJ). Effects of compounds were calculated as: % Inhibition = 100×(IDNJ−Icmpd)/IDNJ, where Icmpd is the DNJ-induced and leak-subtracted current in the presence of a compound or DMSO. Due to lack of effect at the highest dose tested, a dose-response relationship was not examined.

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SGLT2/SGLT1 binding assay: Membrane preparations from human SGLT2/SGLT1-expressing cells were incubated with [³H]-phlorizin (0.5 nM) and Canagliflozin (0.01-10000 nM) at 25°C for 90 minutes. Non-specific binding was determined with excess unlabeled phlorizin. Bound ligands were separated by filtration, and radioactivity was quantified to calculate Ki values [2]
- Sodium-dependent glucose uptake assay: SGLT2-HEK293/SGLT1-HEK293 cells were preincubated with Canagliflozin (0.01-1000 nM) for 20 minutes, then incubated with [¹⁴C]-D-glucose (100 μM) and sodium chloride (140 mM) for 30 minutes. Intracellular radioactivity was measured to determine IC50 values [2]

The effect of canagliflozin on the activity of the glucose transporter 1 (GLUT1) is examined in rat skeletal muscle cell line L6 cells. The culture medium used for the cells is Dulbecco's modified Eagle's medium, which contains 5.6 mM glucose and 10% fetal bovine serum. The cells are seeded in 24-well plates at a density of 3 × 10 5 cells/well and are cultured for 24 hours at 37 °C in an atmosphere of 5% CO2. The cells are pre-incubated with the solutions of Canagliflozin (250 μL, 10 μM) for 5 minutes at room temperature after being 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 Na 2 HPO₄, 10 mM HEPES). 50 μL of 4.5 mM 2-DG (a GLUTs substrate)/3H-2-DG (0.625 μCi) is added to start the transport reaction, which is then incubated for 15 minutes at room temperature. Aspiration of the incubation mixture stops the uptake of 2-DG. The cells are instantly cleaned three times in ice-cold PBS. Radioactivity is measured using liquid scintillation after samples are extracted using 0.3 N NaOH.

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Cell-based Assays [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 were utilized in this study. Cells were seeded into 96-well plates. Cells were 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 was removed, 50 µl of fresh assay buffer with 5 µl of canagliflozin (0.3–300 nM) was 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) was added to the cells and incubated at 37°C for 2 hours. Next, the cells were washed 3 times with 0.15 ml ice-cold phosphate-buffered saline (PBS). After the final wash was aspirated, 50 µl of microscint 20 was added. The plate was counted by TopCount.

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The 2-deoxy-glucose (2-DG) Uptake in L6 Myoblast Cells [1]
Cells from the rat skeletal muscle cell line, L6, was used to test the effect of canagliflozin on glucose transporter 1 (GLUT1) activity. Cells were maintained in Dulbecco's modified Eagle's medium containing 5.6 mM glucose supplemented with 10% fetal bovine serum, were seeded in 24-well plates at a density of 3.0×105 cells/well and cultured for 24 hours in an atmosphere of 5% CO2 at 37°C. Cells were 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 were pre-incubated with the solutions of canagliflozin (250 µl, 10 uM) for 5 minutes at room temperature. The transport reaction was 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 was halted by aspiration of the incubation mixture. Cells were immediately washed 3 times with ice-cold PBS. Samples were extracted with 0.3 N NaOH, and radioactivity was determined by liquid scintillation.

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Renal proximal tubule glucose reabsorption assay: Rat renal proximal tubule cells were cultured on permeable supports, pretreated with Canagliflozin (1-100 nM) for 30 minutes, then exposed to glucose (5 mM) and sodium (140 mM). Transepithelial glucose flux was measured by monitoring glucose concentration in the basolateral medium [1]
- Pancreatic β-cell insulin secretion assay: MIN6 cells were treated with Canagliflozin (0.1-10 μM) plus glucose (5-25 mM) for 2 hours. Insulin levels in supernatants were quantified by ELISA to assess effects on insulin secretion [1]

Dissolved in 0.2% CMC/0.2% Tween 80; 10 mg/kg; oral administration. KK (HF-KK) mice Animals and canagliflozin Administration [1]
\nFour rodent models were 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. Canagliflozin was formulated in 0.5% hydroxypropyl methylcellulose and administrated via oral gavage at 10 ml/kg.

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\n Reduction of Hyperglycemia in Diabetic Rodent Models [1]
\nTo examine the effect of canagliflozin on hyperglycemia, single doses of canagliflozin (0.1, 1, and 10 mg/kg) were administered to overnight-fasted db/db mice. BG levels were monitored at 0, 0.5, 1, 3, 6, and 24 hours after dosing. Canagliflozin was 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 were monitored weekly, and HbA1c, plasma glucose, and insulin levels were determined at the end of the 4-week treatment. An oral glucose tolerance test (OGTT) (2 mg/kg of body weight, given by gavage) was conducted in ZDF rats after 4 weeks of treatment. Blood was 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.

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\n Body Weight Control Studies in Obese Mice and Rats [1]
\nThe effects of canagliflozin on body weight gain were evaluated in DIO mice and ZF rats. DIO mice received a 4-week treatment of canagliflozin at 30 mg/kg. Body weight, food intake, and BG levels were monitored weekly. UGE and indirect calorimetry were conducted in the fourth week of treatment during the compound treatment. In another study, ZF rats were treated with canagliflozin at 3 mg/kg for 3 weeks. Body weight, food intake, and BG were measured weekly during the 19-day treatment period. UGE, body fat, and indirect calorimetric studies were conducted at the end of this study.

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\n Urinary Glucose Excretion (UGE) Study. [1]
\nMale Sprague-Dawley (SD) rats aged 4-5 weeks were used for experiments at 6 weeks of age after acclimation period. The animals were divided into experimental groups matched for body weight (n = 2-3). The compounds were prepared in vehicles as suspension or solution. UGE studies were performed after two-day acclimation period in metabolic cages. The compounds (canagliflozin) or vehicle were orally administered at a dose of 30 mg/kg in 0.2% CMC/0.2% Tween 80. Urine samples were collected for 24 hours using metabolic cages to measure urinary glucose excretion. Urine glucose contents were determined by an enzymatic assay kit (UGLU-L). All animals were allowed free access to a standard pellet diet (CRF1) and tap water.

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\n Single Oral Dosing Study. [1]
\nMale KK/Ta Jcl mice aged 9 weeks were kept on a standard diet (CRF-1; 5.7% (w/w) fat, 3.59 kcal/g), 20-week-old mice were fed with a high-fat diet (60 kcal%) for 4 weeks. The experiment was carried out at the age of 24 weeks. Male C57BL/6N mice aged 11 weeks were also used in this study. The animals were divided into experimental groups matched for body weight and blood glucose levels, which were measured in the fed state on the day of the experiment. The compounds (canagliflozin; 3 mg/kg) or vehicle (0.2% CMC/0.2% Tween 80) were orally administered at a volume of 10 mL/kg. The blood samples were collected from the tail vein before and at 1, 2, 4, 6 and 24 hr after the administration. The blood glucose level was determined using commercially available kits based on the glucose oxidase method. Data are expressed as means ± SEM. Area under the curve for blood glucose levels (AUCglucose 0-6 hr) was calculated by the trapezoidal rule.

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\ndb/db diabetic mouse model: Male db/db mice (8-10 weeks old) were administered Canagliflozin suspended in 0.5% CMC-Na via oral gavage at 1, 3, 10 mg/kg/day for 14 days. Fasting blood glucose, HbA1c, body weight, and insulin levels were measured [1]
\n- ZDF diabetic rat model: Male ZDF rats (10-12 weeks old) were given Canagliflozin (3 mg/kg) dissolved in 0.5% CMC-Na by oral gavage. Urinary glucose excretion, sodium excretion, urine volume, and blood pressure were monitored over 24 hours [1][2]

Absorption, Distribution and Excretion
Bioavailability and Steady-State: The absolute oral bioavailability of canagliflozin averages approximately 65%. Steady-state concentrations are reached after 4 to 5 days with daily doses of 100 mg to 300 mg. Effect of Food on Absorption: Co-administration of canagliflozin with a high-fat meal has no significant effect on its pharmacokinetic parameters. This drug can be taken without food intake. However, because canagliflozin may reduce postprandial plasma glucose excretion by prolonging intestinal glucose absorption, it is recommended to take this drug before the first meal of the day. In healthy subjects, after a single oral dose of radiolabeled canagliflozin, the proportions of canagliflozin or its metabolites detected in feces and urine were as follows: In feces, 41.5% were unmetabolized radiolabeled drug; 7.0% were hydroxylated metabolites; and 3.2% were O-glucuronide metabolites. Approximately 33% of the ingested radiolabeled dose is detected in urine, usually as an O-glucuronide metabolite. Less than 1% of the dose is excreted in urine as unmetabolized drug. The drug is widely distributed throughout the body. The mean steady-state volume of distribution after a single intravenous injection of canagliflozin in healthy subjects is 83.5 L. The clearance of canagliflozin after intravenous injection in healthy subjects is approximately 192 mL/min. Renal clearance of canagliflozin at 100 mg and 300 mg doses ranges from 1.30 to 1.55 mL/min. /Breast Milk/ Canagliflozin is distributed in rat milk; it is unknown whether the drug is distributed in human milk. Canagliflozin is an oral hypoglycemic agent used to treat type 2 diabetes. It works by inhibiting sodium-glucose cotransporter 2, blocking the reabsorption of glucose in the proximal renal tubules. This article describes the biotransformation and distribution of canagliflozin in vivo following a single oral administration of [(14)C]canagliflozin in intact mice and bile duct cannulated (BDC) mice, rats, intact dogs, and humans. Fecal excretion is the primary route of elimination of the radioactive material from the drug in both animals and humans. In BDC mice and rats, most of the radioactive material is excreted via bile. The amount of radioactive material excreted in urine in animals ranges from 1.2% to 7.6% of the administered [(14)C]canagliflozin dose, while in humans it is approximately 33%. The primary metabolic pathway for canagliflozin elimination is oxidation in animals and direct glucuronidation in humans. Unmetabolized canagliflozin is the major component in systemic circulation in all species. In human plasma, the two pharmacologically inactive O-glucuronide conjugates, M5 and M7, account for 19% and 14% of total drug exposure, respectively, and are considered the major human metabolites. In repeated-dose safety studies in mice and rats, plasma concentrations of M5 and M7 were lower than in humans taking the maximum recommended dose of 300 mg canagliflozin. However, bile metabolite analysis in rodents showed significant exposure to M5 and M7 in the livers of mice and rats. The pharmacological inactivity and high water solubility of M5 and M7 support glucuronidation of canagliflozin as a safe detoxification route. The mean absolute oral bioavailability of canagliflozin is approximately 65%. Concomitant administration of canagliflozin with a high-fat meal does not affect its pharmacokinetics; therefore, INVOKANA can be taken with or without food. However, given that INVOKANA can delay intestinal glucose absorption, thereby reducing postprandial glycemic variability, it is recommended to take it before the first meal of the day. Following a single intravenous infusion of canagliflozin in healthy subjects, the mean steady-state volume of distribution was 119 L, indicating extensive tissue distribution. Canagliflozin is extensively bound to plasma proteins (99%), primarily albumin. Protein binding was not correlated with canagliflozin plasma concentration. Plasma protein binding was not significantly altered in patients with impaired renal or hepatic function. Following a single oral dose of [14C] canagliflozin in healthy subjects, 41.5%, 7.0%, and 3.2% of the radioactive dose were recovered in feces, respectively, and excreted as canagliflozin, hydroxylated metabolites, and O-glucuronide metabolites. Enterohepatic circulation of canagliflozin was negligible. Approximately 33% of the radioactive dose was excreted in the urine, primarily as O-glucuronide metabolites (30.5%). Less than 1% of canagliflozin was excreted unchanged in the urine. Renal clearance of canagliflozin at 100 mg and 300 mg doses ranged from 1.30 to 1.55 mL/min. The mean systemic clearance after intravenous administration of canagliflozin in healthy subjects was approximately 192 mL/min.

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Metabolism/Metabolites
Canagliflozin is primarily metabolized via O-glucuronidation.
It is primarily eliminated via glucuronidation by the enzymes UGT1A9 and UGT2B4, producing two inactive O-glucuronide metabolites. In the human body, the oxidative metabolism of canagliflozin via the hepatic cytochrome P44 enzyme CYP3A4 is negligible (approximately 7%). O-glucuronidation is the main metabolic elimination pathway of canagliflozin, primarily carried out by UGT1A9 and UGT2B4, producing two inactive O-glucuronide metabolites. The CYP3A4-mediated (oxidative) metabolism of canagliflozin in the human body is minimal (approximately 7%).
Biological Half-Life
In a clinical study, the terminal half-life of canagliflozin was 10.6 hours in the 100 mg dose group and 13.1 hours in the 300 mg dose group.

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Oral bioavailability: Approximately 78% in rats after oral administration; approximately 83% in dogs after oral administration [2]
-Elimination half-life: 10.2 hours in rats; 16.8 hours in dogs [2]
-Plasma protein binding: 91-94% in human plasma (concentration range: 0.1-10 μg/mL) [2]
-Distribution: Volume of distribution (Vd) in rats = 1.1 L/kg; widely distributed in the kidneys and small intestine [2]
-Metabolism: Mainly metabolized in the liver by CYP3A4 and UDP-glucuronyl transferases (UGTs) into inactive metabolites [2]
-Excretion: 60-65% of the dose is excreted in feces as metabolites; 25-30% is excreted in urine; <5% is excreted unchanged [2]

Toxicity Summary
Identification and Use: Canagliflozin is an oral renal sodium/glucose cotransporter 2 (SGLT2) inhibitor that causes glycosuria and provides a unique mechanism for lowering blood glucose levels in patients with diabetes. Human Exposure and Toxicity: Canagliflozin is used to treat type 2 diabetes. It lowers blood glucose primarily by increasing urinary glucose excretion through inhibition of sodium-glucose cotransporter 2 (SGLT2) in the kidneys. Data from randomized clinical trials lasting up to 52 weeks indicate that canagliflozin is generally well tolerated. The most common adverse reaction is genital fungal infection, occurring in 11–15% of women receiving canagliflozin, compared to 2–4% in women randomized to glimepiride or sitagliptin. In men, the corresponding rates were 8–9% and 0.5–1%, respectively. The incidence of urinary tract infection (UTI) was slightly higher (5–7%) after treatment with canagliflozin compared to placebo (4%). The risk of hypoglycemia associated with canagliflozin is slightly higher than with placebo, but this risk increases significantly when used in combination with insulin or sulfonylureas (SU), in patients with chronic kidney disease (CKD), and in elderly patients. Patients using canagliflozin, especially those with CKD, may experience worsening renal function and hyperkalemia. Canagliflozin offers two major advantages: mild weight loss (average 2-4 kg) and lower blood pressure due to its osmotic diuretic effect. However, the latter may cause orthostatic hypotension and dizziness in susceptible individuals. Another noteworthy adverse effect of canagliflozin is an average 8% increase in plasma low-density lipoprotein cholesterol (LDL-C) levels compared to placebo. Elderly patients may discontinue use of the drug due to adverse effects such as urinary frequency, genital fungal infections, and urinary tract infections. Animal studies: A two-year rat study (at doses of 10, 30, and 100 mg/kg) evaluated the carcinogenicity of canagliflozin. The results showed an increased incidence of pheochromocytoma, renal tubular tumors, and testicular interstitial cell tumors in rats. Interstitial cell tumors were associated with elevated luteinizing hormone levels, while pheochromocytomas were likely associated with glucose malabsorption and calcium homeostasis disturbances. Renal tubular tumors may also be associated with glucose malabsorption. In mice, administration of canagliflozin at doses of 10, 30, or 100 mg/kg did not increase the incidence of tumors. In a toxicity study in pupal rats, researchers administered canagliflozin directly to pupal rats from day 21 (PND 21) to day 90 (PND 90) at doses of 4, 20, 65, or 100 mg/kg. The results showed increased kidney weight in all dose groups, and the incidence and severity of renal pelvis and tubular dilatation increased in a dose-dependent manner. The exposure in the lowest tested dose group was equal to or greater than 0.5 times the maximum clinical dose of 300 mg. The renal pelvis dilatation observed in pupal rats did not completely reverse within a one-month recovery period. In studies of administering canagliflozin to pregnant rats or rabbits during organogenesis, or to mothers from day 6 of gestation (GD 6) to PND 21, and to pups in utero and throughout lactation, no similar effects on developing kidneys were observed. Canagliflozin at doses up to 100 mg/kg (approximately 14 and 18 times the clinical dose of 300 mg for males and females, respectively) had no effect on the ability of rats to mate, reproduce, or maintain offspring, although slight changes were observed in some reproductive parameters at the highest dose (decreased sperm velocity, increased number of abnormal sperm, slightly fewer corpus luteum, reduced implantation sites, and reduced pup number). In the Ames assay, canagliflozin was not mutagenic regardless of metabolic activation. In in vitro mouse lymphoma assays, canagliflozin was mutagenic, but only after metabolic activation. Canagliflozin did not show mutagenicity or chromosome breakage in either the oral micronucleus assay or the oral comet assay in rats.

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Interactions
Sodium-glucose cotransporter 2 (SGLT2) inhibitors reduce hyperglycemia by lowering the renal glucose threshold, thereby increasing urinary glucose excretion. They are considered a novel approach for treating type 2 diabetes. SGLT2 inhibitors have been shown to effectively lower glycated hemoglobin levels without causing hypoglycemia, and have added value in promoting weight loss and lowering blood pressure, whether used as monotherapy or in combination with other hypoglycemic agents. Because SGLT2 inhibitors can be used concomitantly with multiple other drugs, this article reviews the potential drug interactions (DDIs) of three main drugs in this class (dapagliflozin, canagliflozin, and empagliflozin). Most existing studies have been conducted in healthy volunteers and have evaluated the pharmacokinetic interferences of single-dose administration of SGLT2 inhibitors. Exposure to each SGLT2 inhibitor tested (assessed by peak plasma concentration (Cmax) and area under the concentration-time curve (AUC)) was not significantly affected by co-administration with other hypoglycemic agents or commonly used cardiovascular drugs in patients with type 2 diabetes. Conversely, these drugs did not affect the pharmacokinetic parameters of dapagliflozin, canagliflozin, or empagliflozin. Some minor changes were considered clinically insignificant. However, as demonstrated by dapagliflozin and canagliflozin, some drugs that specifically interfere with the metabolic pathways of SGLT2 inhibitors (such as rifampin, uridine diphosphate glucuronyl transferase (UGT) inhibitors or inducers) may cause significant changes in SGLT2 inhibitor exposure. Potential drug interactions warrant further attention in patients with type 2 diabetes receiving long-term SGLT2 inhibitor therapy, especially in debilitated patients taking multiple medications concurrently or with impaired hepatic or renal function.
Digoxin: When co-administered with INVOKANA 300 mg, the AUC and mean peak concentration (Cmax) of digoxin increased by 20% and 36%, respectively. Patients taking INVOKANA and digoxin concurrently should be monitored appropriately.
Concomitant use of canagliflozin with drugs that interfere with the renin-angiotensin-aldosterone system (including angiotensin-converting enzyme (ACE) inhibitors or angiotensin II receptor antagonists) may increase the incidence of symptomatic hypotension. Intravascular volumes should be assessed and corrected before initiating canagliflozin in these patients; after initiation of treatment, patients should be monitored for signs and symptoms of hypotension. These drugs may also cause hyperkalemia in patients with moderate renal impairment. Serum potassium levels should be monitored regularly in patients prone to hyperkalemia due to drug therapy after initiation of canagliflozin.
UGT enzyme inducers: Rifampin: Concomitant use of canagliflozin with rifampin (a non-selective inducer that induces multiple UGT enzymes, including UGT1A9 and UGT2B4) can reduce the area under the curve (AUC) of canagliflozin by 51%. This reduction in canagliflozin exposure may decrease its efficacy. If these UGT inducers (e.g., rifampin, phenytoin sodium, phenobarbital, ritonavir) must be used in combination with INVOKANA (canagliflozin), and if the patient currently tolerates INVOKANA 100 mg once daily, has an eGFR greater than 60 mL/min/1.73 m², and requires additional glycemic control, the dose may be increased to 300 mg once daily. For patients with an eGFR of 45 to less than 60 mL/min/1.73 m² who are receiving UGT inducer therapy and require additional glycemic control, other glycemic treatments should be considered. For more complete interaction data on canagliflozin (6 types), please visit the HSDB records page.

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Acute toxicity: Oral LD50 in rats and mice > 2000 mg/kg [2]
-Subchronic toxicity (oral administration in rats over 28 days): No significant hepatotoxicity or nephrotoxicity was observed at doses up to 30 mg/kg/day; mild transient glycosuria and nauria (physiological effects) [1][2]
-No significant electrolyte abnormalities (potassium, sodium) or renal impairment at therapeutic doses [1]
-Drug interactions: Preclinical studies have shown that CYP3A4 inhibitors (e.g., ketoconazole) can inhibit its effects; no interaction with metformin or insulin [2]

[1]. PLoS One. 2012;7(2):e30555.

[2]. J Med Chem. 2010 Sep 9;53(17):6355-60.

Therapeutic Uses
ClinicalTrials.gov is a registry and results database that lists human clinical studies funded by public and private institutions worldwide. The website is maintained by the National Library of Medicine (NLM) and the National Institutes of Health (NIH). Each record on ClinicalTrials.gov includes summary information about the study protocol, including: the disease or condition; the intervention (e.g., the medical product, behavior, or procedure being studied); the title, description, and design of the study; participation requirements (eligibility criteria); the location of the study; contact information for the study location; and links to relevant information from other health websites, such as the NLM's MedlinePlus (which provides patient health information) and PubMed (which provides citations and abstracts of academic articles in the medical field). Canagliflozin is listed in the database. Invokana (canagliflozin) is indicated as an adjunct to diet and exercise for improving glycemic control in adults with type 2 diabetes. /US Product Label Includes/
Exploratory Treatment of Sodium-Glucose Cotransporter 2 (SGLT2) in the Proximal Tubule: SGLT2 is responsible for the reabsorption of glucose by the kidneys. Therefore, selective inhibition of SGLT2 can lead to significant glycosuria, lower blood glucose, and help weight loss in diabetic patients. In the past year, two SGLT2 inhibitors—canagliflozin and dapagliflozin—have been approved for the treatment of type 2 diabetes. However, in addition to their glycemic effects, these new drugs possess several other properties that provide a theoretical basis for kidney protection. Similar to drugs that block the renin-angiotensin system, SGLT2 inhibitors can also reduce the single glomerular filtration rate (SNGFR) in patients with chronic kidney disease, but the mechanism is quite different. Other potential benefits of SGLT2 inhibition include a moderate reduction in blood pressure and plasma uric acid levels. Furthermore, cell culture studies have shown that glucose uptake in the renal tubule lumen and basolateral membrane promotes the production of extracellular matrix proteins in the proximal tubules. Whether these properties translate into a slowing effect on the progression of chronic kidney disease requires further long-term, dedicated research.
Exploring treatments for diabetes mellitus complicated by hypertension is crucial for reducing cardiovascular mortality and morbidity. Despite improvements in blood pressure control over the past two decades, the blood pressure control rate in patients with type 2 diabetes mellitus (T2DM) remains well below 50%. Recently, a new oral hypoglycemic agent has been approved; sodium-glucose cotransporter 2 (SGLT2) inhibitors work by excreting large amounts of glucose in the urine. Dapagliflozin and canagliflozin are currently approved in the US and Europe, while empagliflozin and iodaggliflozin have completed phase 3 clinical trials. In addition to lowering blood sugar, SGLT2 inhibitors are associated with weight loss and act as osmotic diuretics, thereby lowering blood pressure. Although SGLT2 inhibitors are not yet approved for lowering blood pressure, they may help patients whose blood pressure is 7-10 mmHg below target. Drug Warning: Hypersensitivity reactions (such as generalized urticaria) have been reported with canagliflozin treatment, some of which are quite severe. These reactions typically occur within hours to days after starting canagliflozin. If a hypersensitivity reaction occurs, the medication should be discontinued immediately, appropriate treatment should be administered, and the patient should be closely monitored until symptoms and signs subside. A dose-dependent increase in low-density lipoprotein cholesterol (LDL-C) may occur during canagliflozin treatment. Serum LDL-C concentrations should be monitored during canagliflozin treatment, and such lipid elevations should be managed according to standard treatment protocols. The incidence of hypoglycemia is increased when canagliflozin is used in combination with insulin secretagogues (e.g., sulfonylureas) or insulin compared to sulfonylureas or insulin alone. Therefore, patients receiving canagliflozin may need to reduce the dose of the concurrently used insulin secretagogue or insulin to reduce the risk of hypoglycemia. Canagliflozin may increase the risk of genital fungal infections in men (e.g., balanitis, candidal balanitis) and women (e.g., vulvovaginal candidiasis, vulvovaginal fungal infection, vulvovaginitis). Clinical trials have shown that patients with a history of genital fungal infections and men who have not undergone circumcision are more likely to develop such infections. Patients should be monitored for genital fungal infections, and if such infections occur, appropriate treatment should be administered. For more complete data on drug warnings for canagliflozin (14 in total), please visit the HSDB record page. Pharmacodynamics: This drug increases urinary glucose excretion and lowers the renal glucose threshold (RTG) in a dose-dependent manner. The renal glucose threshold is defined as the minimum blood glucose level required for detectable glucose to be present in the urine. The ultimate result of canagliflozin administration is increased urinary glucose excretion and reduced renal glucose reabsorption, thereby lowering blood glucose concentrations and improving glycemic control. Regarding type 2 diabetes and cardiovascular disease: Patients with type 2 diabetes have an increased risk of cardiovascular events due to the damage to blood vessels and nerves in the cardiovascular system caused by diabetes. In particular, high blood glucose easily leads to pro-atherosclerotic (plaque) lesions in the blood vessels, resulting in a variety of fatal and non-fatal events, including stroke and myocardial infarction. Long-term glycemic control has been proven effective in preventing cardiovascular events such as myocardial infarction and stroke in patients with type 2 diabetes.

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Cagglitazone (JNJ 28431754) is a selective SGLT2 inhibitor used to treat type 2 diabetes mellitus (T2DM)[1][2]
- Its core mechanism is to block SGLT2 in the proximal tubules of the kidney, inhibiting sodium-dependent glucose reabsorption and promoting glycosuria (excretion of glucose in urine), thereby lowering blood glucose levels[1][2]
- It has an insulin-independent hypoglycemic effect and is therefore suitable for patients with T2DM who have insulin resistance[1]
- Other benefits include weight loss (calorie expenditure through glycosuria) and lower blood pressure (through natriuresis and diuresis)[1]
- Its high selectivity for SGLT2 minimizes gastrointestinal reactions. Side effects associated with SGLT1 inhibitors (e.g. diarrhea)[2]
- It is approved for the treatment of type 2 diabetes mellitus on a once-daily dosing regimen due to its long elimination half-life[2]

C24H25FO5S

444.52

444.14

C, 64.85; H, 5.67; F, 4.27; O, 18.00; S, 7.21

842133-18-0

Canagliflozin hemihydrate; 928672-86-0; Canagliflozin-d4; 1997338-61-0; Canagliflozin-d6

24812758

White solid powder

1.4±0.1 g/cm3

642.9±55.0 °C at 760 mmHg

68-72

342.6±31.5 °C

0.0±2.0 mmHg at 25°C

1.639

5.34

4

7

5

31

574

5

O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1C1C=CC(C)=C(CC2=CC=C(C3C=CC(F)=CC=3)S2)C=1

XTNGUQKDFGDXSJ-ZXGKGEBGSA-N

InChI=1S/C24H25FO5S/c1-13-2-3-15(24-23(29)22(28)21(27)19(12-26)30-24)10-16(13)11-18-8-9-20(31-18)14-4-6-17(25)7-5-14/h2-10,19,21-24,26-29H,11-12H2,1H3/t19-,21-,22+,23-,24+/m1/s1

(2S,3R,4R,5S,6R)-2-[3-[[5-(4-fluorophenyl)thiophen-2-yl]methyl]-4-methylphenyl]-6-(hydroxymethyl)oxane-3,4,5-triol

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.62 mM) (saturation unknown) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (5.62 mM) (saturation unknown) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (4.68 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 4: ≥ 2.08 mg/mL (4.68 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 5: ≥ 2.08 mg/mL (4.68 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 6: ≥ 0.5 mg/mL (1.12 mM) (saturation unknown) in 1% DMSO 99% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 7: 0.5% CMC+0.25% Tween 80 : 18 mg/mL

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 (Please use freshly prepared in vivo formulations for optimal results.)