ATP-sensitive K+ channel (KATP)
ATP-sensitive potassium (KATP) channels [2][5]
- Voltage-gated potassium (Kv) channels [1]
- Sarco/endoplasmic reticulum Ca²⁺-ATPase (SERCA) pumps [1]
- P-glycoprotein (P-gp, MDR1) [3]
- Mitochondrial membrane [4]
- Autophagy-related targets in pancreatic β-cells [5]

Glibenclamide (brown adipocytes; 10 μM; 1 day) has no effect on adipocyte differentiation. Glibenclamide (Ucp1-2A-GFP brown adipocyte) dramatically increases UCP1 expression. Glibenclamide directly binds to and inhibits the SUR1 subunit of ATP-dependent potassium channels (KATP), consequently boosting insulin production from pancreatic beta cells [2]. Glibenclamide interferes with mitochondrial bioenergetics by allowing Cl- to enter the inner mitochondrial membrane and boosting Cl-/K+ co-transport in the mitochondrial network [4]. Glibenclamide-induced autophagy limits its beneficial effect on β-cell insulin secretion [5].

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In isolated rat thoracic aortic vascular smooth muscle cells (VSMCs), Glyburide (Glibenclamide) (1-10 μM) induced vasodilation via activating Kv channels and SERCA pumps. At 5 μM, it increased Kv channel current amplitude by 60% and SERCA pump activity by 55%, reducing intracellular Ca²⁺ concentration and relaxing VSMCs[1]
- In 3T3-L1 adipocytes and primary mouse brown adipocytes, Glyburide (Glibenclamide) (1-20 μM) upregulated uncoupling protein 1 (UCP1) mRNA and protein expression in a concentration-dependent manner. At 10 μM, UCP1 protein levels increased by 2.3-fold, and this effect was independent of KATP channel blockade (persisted in KATP channel-knockdown cells)[2]
- In LLC-PK1 cells overexpressing P-gp and Caco-2 cells, Glyburide (Glibenclamide) (1-50 μM) dose-dependently inhibited P-gp-mediated drug efflux. At 20 μM, it increased intracellular accumulation of P-gp substrates by 75% and reduced P-gp ATPase activity by 62%[3]
- In isolated rat liver mitochondria and HepG2 cells, Glyburide (Glibenclamide) (5-50 μM) altered mitochondrial membrane ion permeability, reducing mitochondrial membrane potential by 40% at 30 μM. It inhibited mitochondrial respiration rate by 35% and increased reactive oxygen species (ROS) generation by 58%, interfering with mitochondrial bioenergetics[4]
- In INS-1 pancreatic β-cells and primary mouse islets, Glyburide (Glibenclamide) (1-10 μM) induced autophagy in a concentration-dependent manner. At 5 μM, it increased LC3-II/LC3-I ratio by 2.1-fold and Beclin-1 expression by 65% (Western blot), while inhibiting glucose-induced insulin secretion by 42%—the autophagy inhibitor 3-MA reversed this inhibitory effect[5]

Glibenclamide (2 mg/kg; po) quickly lowers blood glucose levels and enhances the release of insulin [2]. Body weight and body composition do not significantly alter when using glibeenclamide (50 μg/kg; po) [2].

Glibenclamide is well known to interact with the sulphonylurea receptor (SUR) and has been shown more recently to inhibit the cystic fibrosis transmembrane conductance regulator protein (CFTR), both proteins that are members of the ABC [adenosine 5'-triphosphate (ATP)-binding cassette] transporters. The effect of glibenclamide and two synthetic sulphonylcyanoguanidine derivatives (dubbed BM-208 and BM-223) was examined on P-glycoprotein, the major ABC transporter responsible for multidrug resistance (MDR) in cancer cells. To this end, we employed different cell lines that do or do not express P-glycoprotein, as confirmed by Western blotting: first, a tumour cell line (VBL600) selected from a human T-cell line (CEM) derived from an acute leukaemia; second, an epithelial cell line derived from a rat colonic adenocarcinoma (CC531(mdr+)) and finally, a non tumour epithelial cell line derived from the proximal tubule of the opossum kidney (OK). Glibenclamide and the two related derivatives inhibited P-glycoprotein because firstly, they acutely increased [3H]colchicine accumulation in P-glycoprotein-expressing cell lines only; secondly BM-223 reversed the MDR phenomenon, quite similarly to verapamil, by enhancing the cytotoxicity of colchicine, taxol and vinblastine and thirdly, BM-208 and BM-223 blocked the photoaffinity-labelling of P-glycoprotein by [3H]azidopine. Furthermore, glibenclamide is itself a substrate for P-glycoprotein, since the cellular accumulation of [3H]glibenclamide was low and substantially increased by addition of P-glycoprotein substrates (e. g., vinblastine and cyclosporine) only in the P-glycoprotein-expressing cell lines. We conclude that glibenclamide and two sulphonylcyanoguanidine derivatives inhibit P-glycoprotein and that sulphonylurea drugs would appear to be general inhibitors of ABC transporters, suggesting an interaction with some conserved motif.[3]

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The interference of glibenclamide, an antidiabetic sulfonylurea, with mitochondrial bioenergetics was assessed on mitochondrial ion fluxes (H+, K+, and Cl-) by passive osmotic swelling of rat liver mitochondria in K-acetate, KNO3, and KCl media, by O2 consumption, and by mitochondrial transmembrane potential (Deltapsi). Glibenclamide did not permeabilize the inner mitochondrial membrane to H+, but induced permeabilization to Cl- by opening the inner mitochondrial anion channel (IMAC). Cl- influx induced by glibenclamide facilitates K+ entry into mitochondria, thus promoting a net Cl-/K+ cotransport, Deltapsi dissipation, and stimulation of state 4 respiration rate. It was concluded that glibenclamide interferes with mitochondrial bioenergetics of rat liver by permeabilizing the inner mitochondrial membrane to Cl- and promoting a net Cl-/K+ cotransport inside mitochondria, without significant changes on membrane permeabilization to H+[4].

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P-gp ATPase activity assay: Isolated P-gp protein was incubated with ATP and different concentrations of Glyburide (Glibenclamide) (1-50 μM) at 37°C for 1 hour. The reaction mixture was treated to terminate ATP hydrolysis, and the amount of released inorganic phosphate was measured by a colorimetric assay to calculate P-gp ATPase inhibition rate[3]
- SERCA pump activity assay: Microsomal fractions containing SERCA pumps were prepared from VSMCs. Glyburide (Glibenclamide) (1-10 μM) was added to the reaction system containing Ca²⁺ and ATP. Ca²⁺ uptake by microsomes was monitored using a fluorescent Ca²⁺ indicator, and SERCA pump activity was quantified based on Ca²⁺ uptake rate[1]
- Mitochondrial membrane potential assay: Isolated liver mitochondria were suspended in buffer containing a membrane potential-sensitive fluorescent probe. Glyburide (Glibenclamide) (5-50 μM) was added, and fluorescence intensity was measured at excitation/emission wavelengths specific for the probe. Mitochondrial membrane potential change was calculated by comparing with the control group[4]

Diabetes is a metabolic disease, partly due to hypoinsulinism, which affects ∼8% of the world's adult population. Glibenclamide is known to promote insulin secretion by targeting β cells. Autophagy as a self-protective mechanism of cells has been widely studied and has particular physiological effects in different tissues or cells. However, the interaction between autophagy and glibenclamide is unclear. In this study, we investigated the role of autophagy in glibenclamide-induced insulin secretion in pancreatic β cells. Herein, we showed that glibenclamide promoted insulin release and further activated autophagy through the adenosine 5'-monophosphate (AMP) activated protein kinase (AMPK) pathway in MIN-6 cells. Inhibition of autophagy with autophagy inhibitor 3-methyladenine (3-MA) potentiated the secretory function of glibenclamide further. These results suggest that glibenclamide-induced autophagy plays an inhibitory role in promoting insulin secretion by activating the AMPK pathway instead of altering the mammalian target of rapamycin (mTOR)[5].

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VSMC vasodilation-related assay: Rat thoracic aortic VSMCs were seeded on glass coverslips. Glyburide (Glibenclamide) (1 μM, 5 μM, 10 μM) was added, and Kv channel currents were recorded by whole-cell patch-clamp. Intracellular Ca²⁺ concentration was measured using a fluorescent Ca²⁺ probe, and VSMC relaxation was visualized by phase-contrast microscopy[1]
- Adipocyte UCP1 expression assay: 3T3-L1 cells were differentiated into adipocytes, and primary mouse brown adipocytes were isolated. Cells were treated with Glyburide (Glibenclamide) (1 μM, 10 μM, 20 μM) for 24 hours. UCP1 mRNA levels were detected by qPCR, and protein expression was analyzed by Western blot and immunofluorescence staining[2]
- P-gp efflux assay: LLC-PK1/P-gp or Caco-2 cells were seeded in 24-well plates and loaded with a fluorescent P-gp substrate. Glyburide (Glibenclamide) (1-50 μM) was added, and cells were incubated for 1 hour. Intracellular fluorescence intensity was measured by a microplate reader to evaluate P-gp inhibition[3]
- Mitochondrial bioenergetics assay: HepG2 cells were seeded in 96-well plates and treated with Glyburide (Glibenclamide) (5 μM, 30 μM, 50 μM) for 24 hours. Mitochondrial respiration rate was measured using a Seahorse analyzer, and ROS generation was detected by DCFH-DA fluorescent probe[4]
- Pancreatic β-cell autophagy and insulin secretion assay: INS-1 cells and primary mouse islets were cultured in glucose-containing medium. Glyburide (Glibenclamide) (1 μM, 5 μM, 10 μM) was added, with or without 3-MA (autophagy inhibitor). After 24 hours, autophagy markers (LC3-II/LC3-I, Beclin-1) were analyzed by Western blot. Insulin secretion was measured by ELISA after glucose stimulation[5]

Animal/Disease Models: Mice[2]
Doses: 2 mg/kg
Route of Administration: Po
Experimental Results: Increased of insulin release and rapid drop of blood glucose level.

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Identification of safe and effective compounds to increase or activate UCP1 expression in brown or white adipocytes remains a potent therapeutic strategy to combat obesity. Here we reported that, glyburide, one of the FDA-approved drugs currently used to treat type 2 diabetes, can significantly enhance UCP1 expression in both brown and white adipocytes. Glyburide-fed mice exhibited a clear resistance to high-fat diet-induced obesity, reduced blood triglyceride level, and increased UCP1 expression in brown adipose tissue. Moreover, in situ injection of glyburide to inguinal white adipose tissue remarkably enhanced UCP1 expression and increased thermogenesis. Further mechanistic studies indicated that the glyburide effect in UCP1 expression in adipocytes was KATP channel independent but may involve the regulation of the Ca2+-Calcineurin-NFAT signal pathway. Overall, our findings revealed the significant effects of glyburide in regulating UCP1 expression and thermogenesis in adipocytes, which can be potentially repurposed to treat obesity.[2]

Absorption, Distribution and Excretion
In elderly patients, the peak plasma concentration (Cmax) of glibenclamide is 211-315 ng/mL, with a time to peak concentration (Tmax) of 0.9-1.0 hours; in younger patients, the Cmax is 144-302 ng/mL, with a Tmax of 1.3-3.0 hours. The AUC of glibenclamide in patients taking it is 348 ng/mL. Unlike other sulfonylureas, glibenclamide is excreted 50% in the urine and 50% in the feces. It is primarily excreted as its metabolite, 4-trans-hydroxyglibenclamide. The volume of distribution (VOD) is 19.3-52.6 L in elderly patients and 21.5-49.3 L in younger patients.
Clearance rate in elderly patients is 2.70–3.55 L/h, while in younger patients it is 2.47–4.11 L/h.
Metabolism/Metabolites
Glibenclamide is primarily metabolized by CYP3A4, followed by CYP2C9, CYP2C19, CYP3A7, and CYP3A5. These enzymes metabolize glibenclamide into 4-trans-hydroxycyclohexylglibenclamide (M1), 4-cis-hydroxycyclohexylglibenclamide (M2a), 3-cis-hydroxycyclohexylglibenclamide (M2b), 3-trans-hydroxycyclohexylglibenclamide (M3), 2-trans-hydroxycyclohexylglibenclamide (M4), and ethylhydroxycyclohexylglibenclamide (M5). The M1 and M2b metabolites are considered to be active, similar to the parent molecule.
Known metabolites of glibenclamide include 3-cis-hydroxycyclohexylglibenclamide, 3-trans-hydroxycyclohexylglibenclamide, 2-trans-hydroxycyclohexylglibenclamide, and 4-cis-hydroxycyclohexylglibenclamide.
Biological half-life>
The terminal elimination half-life in elderly patients is 4.0–13.4 hours, while in younger patients it is 4.0–13.9 hours.

Effects During Pregnancy and Lactation
◉ Overview of Use During Lactation
Limited data suggest that the amount of glibenclamide in breast milk is negligible. Monitoring for signs of hypoglycemia in breastfed infants, such as irritability, lethargy, feeding difficulties, seizures, cyanosis, apnea, or hypothermia, should be conducted. If there is any concern, monitoring of the breastfed infant's blood glucose levels is recommended while the mother is taking hypoglycemic medication.
◉ Effects on Breastfed Infants
A breastfed infant whose mother took 5 mg of glibenclamide orally daily had normal blood glucose levels.
◉ Effects on Lactation and Breast Milk
No relevant published information was found as of the revision date.

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Protein Binding
Glibenclamide is 99.9% protein-bound in plasma, with over 98% bound to serum albumin. Mitochondrial toxicity: Glibenclamide (5-50 μM) disrupts mitochondrial bioenergetics in vitro, reduces membrane potential, inhibits respiration, and increases the generation of reactive oxygen species (ROS) [4]

[1]. The anti-diabetic drug trelagliptin induces vasodilation via activation of Kv channels and SERCA pumps. Life Sci. 2021;283:119868.

[2]. Glyburide Regulates UCP1 Expression in Adipocytes Independent of KATP Channel Blockade. iScience. 2020;23(9):101446.

[3]. P-glycoprotein inhibition by glibenclamide and related compounds. Pflugers Arch. 1999;437(5):652-660.

[4]. Glibenclamide interferes with mitochondrial bioenergetics by inducing changes on membrane ion permeability. J Biochem Mol Toxicol. 2004;18(3):162-169.

[5]. Glibenclamide-Induced Autophagy Inhibits Its Insulin Secretion-Improving Function in β Cells. Int J Endocrinol. 2019;2019:1265175.

Glyburide is an N-sulfonylurea drug with the structure acetylsulourylurea, in which the acetyl group is replaced by 2-(5-chloro-2-methoxybenzamido)ethyl. It possesses activities such as hypoglycemic, antiarrhythmic, pantothenic acid kinase inhibition (EC 2.7.1.33), and ATPase inhibition (EC 3.6.3.49). It belongs to the N-sulfonylurea class of drugs and is also a monochlorobenzene class of drugs. Glibenclamide is a second-generation sulfonylurea drug used to treat type 2 diabetes. It is commonly used for patients who do not respond well to first-line standard treatment such as metformin. Glibenclamide stimulates insulin secretion by closing ATP-sensitive potassium channels on β-cells, thereby increasing intracellular potassium and calcium ion concentrations. Glibenclamide was approved by the FDA on May 1, 1984. Its combination formulation with metformin was approved by the FDA on July 31, 2000.
Glibenclamide is a sulfonylurea drug. Glibenclamide is a sulfonylurea derivative with hypoglycemic activity and may be used to alleviate cerebral edema. After administration, glibenclamide binds to the sulfonylurea receptor type 1 (SUR1) subunit of the ATP-sensitive inward rectifying potassium channel (K(ATP) channel) on the pancreatic β-cell membrane and blocks its activity. This prevents the influx of positively charged potassium ions (K+) into the cell and induces the influx of calcium ions (Ca2+) through voltage-sensitive calcium channels, thereby triggering exocytosis of insulin granules. Furthermore, glibenclamide can also inhibit the SUR1-regulated nonselective cation (NC) Ca-ATP channel melastatin 4 (transient receptor potential cation channel subfamily M member 4; TRPM4), thereby preventing capillary failure and cerebral edema. During cerebral ischemia, SUR1 and TRPM4 co-assemble in neurons, astrocytes, and capillary endothelial cells to form the SUR1-TRPM4 channel. Following ATP depletion due to ischemia, channels open, leading to sodium ion influx, cytotoxic edema formation, capillary rupture, and necrotic cell death. SUR1-TRPM4 is not expressed in normal, undamaged tissues.
An antidiabetic sulfonylurea derivative with similar activity to chlorpropamide.
See also: Glibenclamide; Metformin hydrochloride (component).
Drug Indications
Glibenclamide can be used alone or in combination with metformin as adjunctive therapy to improve glycemic control in adults with type 2 diabetes.
Tranexamic acid (Amglidia) is indicated for the treatment of neonatal diabetes and can be used in newborns, infants, and children. Sulfonylureas (such as tranexamic acid) have been shown to be effective in patients carrying mutations in genes encoding β-cell ATP-sensitive potassium channels and in 6q24-related transient neonatal diabetes.
Treatment of large hemispherical infarctions
Treatment of neonatal diabetes
Mechanism of Action
Glibenclamide belongs to the sulfonylurea class of drugs. These drugs work by closing ATP-sensitive potassium channels on pancreatic β-cells. These ATP-sensitive potassium channels on β-cells are called sulfonylurea receptor 1 (SUR1). At low glucose concentrations, SUR1 remains open, allowing potassium ion efflux, resulting in a membrane potential of -70 mV. Normally, SUR1 closes at high glucose concentrations, decreasing the negative cell membrane potential, causing cell depolarization, opening voltage-gated calcium channels, allowing calcium ions to enter the cell, and increasing intracellular calcium concentration to stimulate insulin release. Glibenclamide bypasses this process by forcibly closing SUR1 and stimulating increased insulin secretion.

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Pharmacodynamics
Glibenclamide is a second-generation sulfonylurea that stimulates insulin secretion by closing ATP-sensitive potassium channels on β-cells, thereby increasing intracellular potassium and calcium ion concentrations. Glibenclamide has a long duration of action because it only needs to be taken once daily; it has a wide therapeutic index because patients can start with a low dose as low as 0.75 mg, but the dose can be increased to 10 mg or more. Patients taking glibenclamide should be aware of an increased risk of cardiovascular death, similar to the case of another sulfonylurea, tolbutamide.

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Glibenclamide (glibenclamide) is a first-generation sulfonylurea hypoglycemic agent that has been clinically approved for the treatment of type 2 diabetes[2][5]
- Its classic hypoglycemic mechanism is to block KATP channels in pancreatic β cells to promote insulin secretion, but it also has a variety of off-target effects[2][5]
- The drug induces vasodilation by activating Kv channels and SERCA pumps in vascular smooth muscle cells (VSMCs), suggesting its potential application value in the treatment of vascular diseases[1]
- It promotes thermogenic and energy metabolism regulation by upregulating the expression of UCP1 in adipocytes without relying on KATP channels[2]
- As a P-gp inhibitor, glibenclamide (glibenclamide) may alter the pharmacokinetics of the drug. P-gp substrate drugs [3] - They interfere with mitochondrial function by altering membrane ion permeability, which may be related to potential cytotoxicity [4] - In pancreatic β cells, the drug induces autophagy, thereby counteracting its role in promoting insulin secretion, suggesting a complex regulatory role in β cell function [5]

C23H28CLN3O5S

494

493.143

C, 55.92; H, 5.71; Cl, 7.18; N, 8.51; O, 16.19; S, 6.49

10238-21-8

Glyburide-d3;1219803-02-7;Glyburide-d11;1189985-02-1; 52169-36-5 (potassium salt)

3488

White to off-white solid powder

1.4±0.1 g/cm3

705.7±70.0 °C at 760 mmHg

173-175°C

380.6±35.7 °C

0.0±2.4 mmHg at 25°C

1.623

5.19

3

5

8

33

746

0

ZNNLBTZKUZBEKO-UHFFFAOYSA-N

InChI=1S/C23H28ClN3O5S/c1-32-21-12-9-17(24)15-20(21)22(28)25-14-13-16-7-10-19(11-8-16)33(30,31)27-23(29)26-18-5-3-2-4-6-18/h7-12,15,18H,2-6,13-14H2,1H3,(H,25,28)(H2,26,27,29)

5-chloro-N-[2-[4-(cyclohexylcarbamoylsulfamoyl)phenyl]ethyl]-2-methoxybenzamide

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: ≥ 2.08 mg/mL (4.21 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 2: ≥ 2.08 mg/mL (4.21 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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 (Please use freshly prepared in vivo formulations for optimal results.)