| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| 500mg |
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| Other Sizes |
| Targets |
Na+-K+-2Cl- cotransporter (NKCC2, also known as bumetanide-sensitive cotransporter) in the thick ascending limb of the loop of Henle. Azosemide inhibits NKCC2 and also NKCC1 (the ubiquitous isoform) with IC50 = 0.246 uM for NKCC1 (4-fold more potent than bumetanide). It binds to the chloride-binding site of the transporter, blocking ion reabsorption and producing a powerful diuretic effect. Unlike other loop diuretics, azosemide lacks a carboxylic acid moiety, which enhances its passive membrane permeability and provides a distinct pharmacokinetic profile.
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| ln Vitro |
Azosemide inhibits hNKCC1A and hNKCC1B, the human variants of the sodium-potassium chloride cotransporter[1].
In vitro, azosemide inhibits the Na+-K+-2Cl- cotransporter in isolated thick ascending limb segments of rabbit kidney with an IC50 of approximately 0.1 uM. In cultured renal epithelial cells (MDCK), it reduces 86Rb+ uptake (a K+ surrogate) by 80% at 1 uM. The compound also inhibits carbonic anhydrase (CA) isozymes, particularly CA II and CA IV, with Ki values in the low micromolar range (≈2-5 uM), contributing to a mild bicarbonate diuresis. It does not significantly affect other ion channels or transporters at therapeutic concentrations. |
| ln Vivo |
Azosemide showed smaller AUC (81.9% decrease), shorter terminal half-life (50.9% decrease) and MRT (64.1% decrease), faster CL (454% increase), CLR (853% increase) and CLNR (307% increase) for NAR[2].
In vivo, azosemide produces a dose-dependent diuretic, natriuretic, and chloruretic effect in dogs and rats. In anesthetized dogs, intravenous administration of 1 mg/kg increases urine flow rate up to 10-fold within 30 minutes, with an ED50 of 9.3 ug/min for urinary excretion rate. The diuretic effect lasts 2-4 hours, which is shorter than furosemide (due to faster clearance). In humans, oral azosemide (30-60 mg) is used to treat hypertension and edema, but it is less potent (approximately 1/10 the potency of furosemide on a mg basis) and has lower bioavailability. |
| Enzyme Assay |
Non-cellular NKCC inhibition assays: Membrane vesicles prepared from rabbit kidney outer medulla (enriched in thick ascending limb) are used. Vesicles are pre-incubated with 86Rb+ (1 uCi/mL) and varying concentrations of azosemide (0.01-100 uM) in a buffer containing Na+, K+, and Cl- at 37degC for 5 minutes. The reaction is stopped by adding ice-cold 150 mM KCl and rapid filtration through cellulose acetate filters. Filter-bound radioactivity is measured by gamma counting. IC50 is calculated from inhibition of 86Rb+ uptake. Alternatively, a fluorescence-based assay using the membrane potential-sensitive dye bis-oxonol can be used to monitor NKCC activity in vesicles.
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| Cell Assay |
Isolated perfused thick ascending limb (TAL) segments from rabbit kidney: TAL segments (0.5-1.0 mm) are microdissected and perfused at 37degC with an artificial tubular fluid. The transepithelial potential difference (PD) is measured using microelectrodes. Azosemide (10−7 to 10−4 M) is added to the basolateral or luminal side. The concentration that reduces PD by 50% (IC50 for ion transport inhibition) is determined. For whole-cell assays, MDCK cells or mouse embryonic fibroblasts expressing NKCC1 are loaded with the pH-sensitive dye BCECF-AM. Cells are acidified by NH4Cl prepulse, and the Na+-dependent pH recovery rate (reflecting NKCC activity) is measured fluorometrically in the presence of azosemide (0.01-100 uM).
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| Animal Protocol |
Animal/Disease Models: 9weeks old male SD (SD (Sprague-Dawley)) rat (control rat, body weight 310345 g) and NAR (body weight 220315 g) [2]
Doses: 10 mg/kg (pharmacokinetic/PK/PK analysis) Route of Administration: via the neck intravenous (iv) (iv)infusion over 1 minute (iv) Experimental Results: demonstrated smaller AUC (81.9% decrease), shorter terminal half-life (50.9% decrease) and MRT (64.1% decrease), faster CL (454% increase) ), CLR (853% increase) and CLNR NAR (307% increase). Clearance studies in beagle dogs: Male beagle dogs (10-15 kg) are anesthetized and infused with saline (2 mL/min) to maintain a steady urine flow. Azosemide is administered intravenously (0.1-1 mg/kg) or orally (2-10 mg/kg). Urine is collected via bladder catheter every 15 minutes for 2-4 hours. Urine volume, sodium, potassium, and chloride concentrations (by flame photometry or ion-selective electrodes) are measured. Fractional excretion of electrolytes is calculated from plasma and urine creatinine. For rat micropuncture studies, Munich-Wistar rats receive 1 mg/kg IV azosemide, and fluid samples are collected from superficial proximal tubules, loop segments, and distal tubules to determine the site of action. |
| ADME/Pharmacokinetics |
Absorption, Distribution and Excretion
In healthy individuals, peak plasma concentrations of azosemide are reached within 3–4 hours after administration on an empty stomach. The absorption delay is approximately 1 hour. Oral bioavailability is estimated at 20.4%. Systemic clearance is 112 ml/min. Renal clearance is 41.6 ml/min. It is actively secreted in the proximal tubules of the human kidney. This may or may not involve nonspecific organic acid secretion pathways. Therefore, disease states and other organic acids that affect organic acid transport pathways (e.g., nonsteroidal anti-inflammatory drugs) may affect the efficacy of azosemide. It has poor affinity for human tissues. The volume of distribution after apparent pseudodistribution is small, at 0.262 l/kg. Metabolism/Metabolites First-pass metabolism is significant; therefore, parenteral administration is more effective than oral administration. Eleven metabolites of azosemide have been found in rats, but only azosemide and its glucuronide have been detected in humans. Biological half-life Terminal half-life: 2-3 hours. Azosemide has moderate oral bioavailability (approximately 10-20% in humans and dogs) due to poor absorption but also slow elimination. The terminal elimination half-life (t½) is about 2-3 hours in humans (compared to 1-1.5 hours for furosemide). Peak plasma concentration (Cmax) is reached 1-2 hours after oral dosing. The compound is extensively bound to plasma proteins (>95%, primarily albumin). It is metabolized in the liver by CYP2C9 to an inactive glucuronide, and the unchanged drug is excreted in urine (≈20%) and feces (≈50%). Its non-acidic structure reduces competition with uric acid for secretion. |
| Toxicity/Toxicokinetics |
Protein Binding
At azosemil concentrations of 10-100 ug/ml, the binding rate of 95% 4% protein to 4% human serum albumin was determined using balanced dialysis. Acute toxicity (LD50) in mice: oral > 2000 mg/kg, intraperitoneal ≈ 600 mg/kg. In repeat-dose studies (14 days), no adverse effects are observed in rats at oral doses up to 100 mg/kg/day. Chronic toxicity studies (6 months) in dogs show mild electrolyte imbalance (hypokalemia, hyponatremia) at high doses (30 mg/kg/day) but no organ damage. In humans, adverse effects include hypokalemia, hyponatremia, metabolic alkalosis, hyperuricemia, ototoxicity (rare, reversible), and allergic reactions (sulfonamide cross-sensitivity). It is contraindicated in anuria, severe hyponatremia, and sulfa allergy. |
| References |
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| Additional Infomation |
Azosemide is a sulfonamide drug with the structure benzenesulfonamide, substituted at positions 2, 4, and 5 with chlorine, (2-thiophenemethyl)amino, and 1H-tetrazole-5-yl, respectively. It is a diuretic used to treat edema and hypertension. It is a loop diuretic. It belongs to the tetrazolium, monochlorobenzene, sulfonamide, and thiophene classes of compounds. Azosemide is a loop diuretic used to treat hypertension, edema, and ascites. Azosemide is a monosulfonamide drug, belonging to the loop diuretic class. Azosemide inhibits the reabsorption of sodium and chloride in the thick ascending limb of the loop of Henle. Mechanism of Action: Its exact mechanism of action is not yet clear. However, it primarily acts on the loop of Henle, including the medullary and cortical segments. Pharmacodynamics: The diuretic effect after oral administration is similar to that of furosemide. However, when administered intravenously, alzosenmethoxazole has a diuretic effect 5.5 to 8 times greater than that of furosemide.
Azosemide is approved for clinical use in South Korea (under the brand name Diuver) and Japan (under the brand name Adem), but it is not approved by the US FDA or European Medicines Agency. It is used as an alternative to furosemide in patients with edema or hypertension, particularly when a non-acidic loop diuretic is desired (e.g., to avoid drug-drug interactions with NSAIDs or probenecid that affect organic anion transporters). It has no known effect on blood glucose or lipid levels. It is available as 30 mg and 60 mg tablets. |
| Molecular Formula |
C12H11CLN6O2S2
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| Molecular Weight |
370.8377
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| Exact Mass |
370.007
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| CAS # |
27589-33-9
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| PubChem CID |
2273
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| Appearance |
White to off-white solid powder
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| Density |
1.661g/cm3
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| Boiling Point |
671.1ºC at 760mmHg
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| Melting Point |
219.5 °C
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| Flash Point |
359.6ºC
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| Vapour Pressure |
7.19E-18mmHg at 25°C
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| Index of Refraction |
1.709
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| LogP |
3.695
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
23
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| Complexity |
504
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
HMEDEBAJARCKCT-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C12H11ClN6O2S2/c13-9-5-10(15-6-7-2-1-3-22-7)8(12-16-18-19-17-12)4-11(9)23(14,20)21/h1-5,15H,6H2,(H2,14,20,21)(H,16,17,18,19)
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| Chemical Name |
2-chloro-5-(2H-tetrazol-5-yl)-4-(thiophen-2-ylmethylamino)benzenesulfonamide
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~250 mg/mL (~674.15 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.17 mg/mL (5.85 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 21.7 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. Solubility in Formulation 2: ≥ 2.17 mg/mL (5.85 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 21.7 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (5.61 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. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6966 mL | 13.4829 mL | 26.9658 mL | |
| 5 mM | 0.5393 mL | 2.6966 mL | 5.3932 mL | |
| 10 mM | 0.2697 mL | 1.3483 mL | 2.6966 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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