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Glyburide (formerly HB419; HB420; Glibenclamide; Micronase; Diabeta; Maninil; Micronase; Neogluconin) is a selective blocker of vascular ATP-sensitive K+ channels (KATP) that has been approved as an antidiabetic sulfonylurea drug used for the treatment of type 2 diabetes. Its actions are similar to those of chlorpropamide that can potentially be used to decrease cerebral edema.
Physicochemical Properties
| Molecular Formula | C23H28CLN3O5S | |
| Molecular Weight | 494 | |
| Exact Mass | 493.143 | |
| Elemental Analysis | C, 55.92; H, 5.71; Cl, 7.18; N, 8.51; O, 16.19; S, 6.49 | |
| CAS # | 10238-21-8 | |
| Related CAS # | Glyburide-d3;1219803-02-7;Glyburide-d11;1189985-02-1; 52169-36-5 (potassium salt) | |
| PubChem CID | 3488 | |
| Appearance | White to off-white solid powder | |
| Density | 1.4±0.1 g/cm3 | |
| Boiling Point | 705.7±70.0 °C at 760 mmHg | |
| Melting Point | 173-175°C | |
| Flash Point | 380.6±35.7 °C | |
| Vapour Pressure | 0.0±2.4 mmHg at 25°C | |
| Index of Refraction | 1.623 | |
| LogP | 5.19 | |
| Hydrogen Bond Donor Count | 3 | |
| Hydrogen Bond Acceptor Count | 5 | |
| Rotatable Bond Count | 8 | |
| Heavy Atom Count | 33 | |
| Complexity | 746 | |
| Defined Atom Stereocenter Count | 0 | |
| InChi Key | ZNNLBTZKUZBEKO-UHFFFAOYSA-N | |
| InChi Code | 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) | |
| Chemical Name | 5-chloro-N-[2-[4-(cyclohexylcarbamoylsulfamoyl)phenyl]ethyl]-2-methoxybenzamide | |
| Synonyms |
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| HS Tariff Code | 2934.99.9001 | |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
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] |
| ln Vitro |
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]. 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] |
| ln Vivo | 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]. |
| Enzyme Assay |
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] 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]. 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] |
| Cell Assay |
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]. 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 Protocol |
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. 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] |
| ADME/Pharmacokinetics |
Absorption, Distribution and Excretion Elderly patients taking glyburide reached a Cmax of 211-315ng/mL with a Tmax of 0.9-1.0h, while younger patients reached a Cmax of 144-302ng/mL with a Tmax of 1.3-3.0h. Patients taking glyburide have and AUC of 348ngh/mL. Unlike other sulfonylureas, glyburide is 50% excreted in the urine and 50% in the feces. Glyburide is mainly excreted as the metabolite 4-trans-hydroxyglyburide. Elderly patients have a volume of distribution of 19.3-52.6L, while younger patients have a volume of distribution of 21.5-49.3L. Elderly patients have a clearance of 2.70-3.55L/h, while younger patients have a clearance of 2.47-4.11L/h. Metabolism / Metabolites Glyburide is metabolized mainly by CYP3A4, followed by CYP2C9, CYP2C19, CYP3A7, and CYP3A5. These enzymes metabolize glyburide to 4-trans-hydroxycyclohexyl glyburide (M1), 4-cis-hydroxycyclohexyl glyburide (M2a), 3-cis-hydroxycyclohexyl glyburide (M2b), 3-trans-hydroxycyclohexyl glyburide (M3), 2-trans-hydroxycyclohexyl glyburide (M4), and ethylhydroxycyclohexyl glyburide (M5). The M1 and M2b metabolites are considered active, along with the parent molecule. Glyburide has known human metabolites that include 3-cis-Hydroxycyclohexyl glyburide, 3-trans-Hydroxycyclohexyl glyburide, 2-trans-hydroxycyclohexyl glyburide, and 4-cis-hydroxycyclohexyl glyburide. Biological Half-Life Elderly patients have a terminal elimination half life of 4.0-13.4h, while younger patients have a terminal elimination half life of 4.0-13.9h. |
| Toxicity/Toxicokinetics |
Effects During Pregnancy and Lactation ◉ Summary of Use during Lactation Limited data indicate that the levels of glyburide in milk are negligible. Monitor breastfed infants for signs of hypoglycemia such as jitteriness, excessive sleepiness, poor feeding, seizures cyanosis, apnea, or hypothermia. If there is concern, monitoring of the breastfed infant's blood glucose is advisable during maternal therapy with hypoglycemic agents. ◉ Effects in Breastfed Infants The blood glucose level was normal in one breastfed infant whose mothers was taking oral glyburide 5 mg daily. ◉ Effects on Lactation and Breastmilk Relevant published information was not found as of the revision date. Protein Binding Glyburide is 99.9% bound to protein in plasma with >98% accounted for by binding to serum albumin. Mitochondrial toxicity: Glyburide (Glibenclamide) (5-50 μM) disrupted mitochondrial bioenergetics in vitro, reducing membrane potential, inhibiting respiration, and increasing ROS generation[4] |
| References |
[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. |
| Additional Infomation |
Glyburide is an N-sulfonylurea that is acetohexamide in which the acetyl group is replaced by a 2-(5-chloro-2-methoxybenzamido)ethyl group. It has a role as a hypoglycemic agent, an anti-arrhythmia drug, an EC 2.7.1.33 (pantothenate kinase) inhibitor and an EC 3.6.3.49 (channel-conductance-controlling ATPase) inhibitor. It is a N-sulfonylurea and a member of monochlorobenzenes. Glyburide is a second generation sulfonylurea used to treat patients with diabetes mellitus type II. It is typically given to patients who cannot be managed with the standard first line therapy, [metformin]. Glyburide stimulates insulin secretion through the closure of ATP-sensitive potassium channels on beta cells, raising intracellular potassium and calcium ion concentrations. Glyburide was granted FDA approval on 1 May 1984. A formulation with metformin was granted FDA approval on on 31 July 2000. Glyburide is a Sulfonylurea. Glyburide is a sulfonamide urea derivative with antihyperglycemic activity that can potentially be used to decrease cerebral edema. Upon administration, glyburide binds to and blocks the sulfonylurea receptor type 1 (SUR1) subunit of the ATP-sensitive inwardly-rectifying potassium (K(ATP)) channels on the membranes of pancreatic beta cells. This prevents the inward current flow of positively charged potassium (K+) ions into the cell, and induces a calcium ion (Ca2+) influx through voltage-sensitive calcium channels, which triggers exocytosis of insulin-containing granules. In addition, glyburide also inhibits 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 brain swelling. SUR1-TRPM4 channels are formed by co-assembly of SUR1 with TRPM4 in neurons, astrocytes, and capillary endothelium during cerebral ischemia. Upon ischemia-induced ATP depletion, channels open which results in sodium influx, cytotoxic edema formation, capillary fragmentation and necrotic cell death. SUR1-TRPM4 is not expressed in normal, uninjured tissues. An antidiabetic sulfonylurea derivative with actions like those of chlorpropamide See also: Glyburide; Metformin Hydrochloride (component of). Drug Indication Glyburide is indicated alone or as part of combination product with metformin, as an adjunct to diet and exercise, to improve glycemic control in adults with type 2 diabetes mellitus. Amglidia is indicated for the treatment of neonatal diabetes mellitus, for use in newborns, infants and children. Sulphonylureas like Amglidia have been shown to be effective in patients with mutations in the genes coding for the β-cell ATP-sensitive potassium channel and chromosome 6q24-related transient neonatal diabetes mellitus. Treatment of large hemispheric infarction Treatment of neonatal diabetes mellitus Mechanism of Action Glyburide belongs to a class of drugs known as sulfonylureas. These drugs act by closing ATP-sensitive potassium channels on pancreatic beta cells. The ATP-sensitive potassium channels on beta cells are known as sulfonylurea receptor 1 (SUR1). Under low glucose concentrations, SUR1 remains open, allowing for potassium ion efflux to create a -70mV membrane potential. Normally SUR1 closes in response to high glucose concentrations, the membrane potential of the cells becomes less negative, the cell depolarizes, voltage gated calcium channels open, calcium ions enter the cell, and the increased intracellular calcium concentration stimulates the release of insulin containing granules. Glyburide bypasses this process by forcing SUR1 closed and stimulating increased insulin secretion. Pharmacodynamics Glyburide is a second generation sulfonylurea that stimulates insulin secretion through the closure of ATP-sensitive potassium channels on beta cells, raising intracellular potassium and calcium ion concentrations. Glibenclamide has a long duration of action as it is given once daily, and a wide therapeutic index as patients are started at doses as low as 0.75mg but that can increase as high as 10mg or more. Patients taking glyburide should be cautioned regarding an increased risk of cardiovascular mortality as seen with tolbutamide, another sulfonylurea. Glyburide (Glibenclamide) is a first-generation sulfonylurea antidiabetic drug clinically approved for the treatment of type 2 diabetes mellitus[2][5] - Its classic hypoglycemic mechanism involves blocking pancreatic β-cell KATP channels to promote insulin secretion, but it also exerts multiple off-target effects[2][5] - The drug induces vasodilation via activating Kv channels and SERCA pumps in VSMCs, suggesting potential applications in vascular disorders[1] - It upregulates UCP1 expression in adipocytes independent of KATP channels, contributing to thermogenesis and energy metabolism regulation[2] - As a P-gp inhibitor, Glyburide (Glibenclamide) may alter the pharmacokinetics of P-gp substrate drugs[3] - It interferes with mitochondrial function by changing membrane ion permeability, which may be related to potential cellular toxicity[4] - In pancreatic β-cells, the drug induces autophagy that counteracts its insulin secretion-improving effect, indicating a complex regulatory role in β-cell function[5] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
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. 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0243 mL | 10.1215 mL | 20.2429 mL | |
| 5 mM | 0.4049 mL | 2.0243 mL | 4.0486 mL | |
| 10 mM | 0.2024 mL | 1.0121 mL | 2.0243 mL |