Glimepiride (HOE-490; HOE490; Amaryl; Glimepiridum; Amarel; Glimepirida; Roname), a third generation and medium-to-long acting sulfonylurea compound, is a potent Kir6.2/SUR inhibitor with potential antidiabetic activity. It inhibits SUR1, SUR2A and SUR2B with IC50s of 3.0 nM, 5.4 nM, and 7.3 nM. It was approved for use in the treatment of type 2 diabetes mellitus. The mechanism of action of Glimepiride is to increase the release of insulin from pancreatic beta cells. In addition, glimepiride increases the activity of intracellular insulin receptors. Glimepiride increases osteoblast proliferation and differentiation, which is thought to be related to its ability to activate the PI3K and Akt pathway.

Glimepiride (Hoe 490) is a new sulfonylurea. After oral administration of Hoe 490 to rabbits, blood glucose was lowered 3.5 times more than after glibenclamide (HB 419) and after intravenous administration, 2.5 times more. This superiority in efficacy was demonstrated by onset, maximum and duration of action. In rats, intravenous and oral Hoe 490 has a much shorter effect on blood glucose than HB 419, but the initial effect of Hoe 490 orally was up to 6 times and i.v. up to 2 times stronger than that of HB 419. In dogs, oral and intravenous Hoe 490 had a considerably longer blood glucose-lowering effect than HB 419. However, the effect of intravenous Hoe 490 was only half as intense as that of HB 419 in the first hours after treatment and the effect of oral Hoe 490 was initially stronger and thereafter temporarily distinctly weaker than that of HB 419. The more rapid decrease in blood glucose in the dog after oral administration of Hoe 490 was accompanied by a correspondingly earlier and higher plasma insulin increase. In accordance with the less intense initial blood glucose decrease in the dog after intravenous Hoe 490 there was a weaker and slower rise and faster drop of plasma insulin. The long action of oral and intravenous Hoe 490 in the dog can, however, not be sufficiently explained by the plasma insulin values. In the isolated rat pancreas perfused with glucose-free medium, HB 419 released glucagon beside insulin and somatostatin. The threshold concentration for the glucagon secretion was lower as those for the insulin and somatostatin release [1].

Physicochemical Properties

Molecular Formula	C24H34N4O5S
Molecular Weight	490.62
Exact Mass	490.224
Elemental Analysis	C, 58.75; H, 6.99; N, 11.42; O, 16.31; S, 6.54
CAS #	93479-97-1
Related CAS #	Glimepiride-d5;1028809-90-6; Glimepiride-d4-1; 1131981-29-7; 119018-30-3 (urethane); 119018-29-0 (sulfonamide); 93479-97-1
PubChem CID	3476
Appearance	White to off-white solid powder
Density	1.3±0.1 g/cm3
Boiling Point	677.0±65.0 °C at 760 mmHg
Melting Point	212.2-214.5 °C
Flash Point	363.2±34.3 °C
Vapour Pressure	0.0±2.2 mmHg at 25°C
Index of Refraction	1.628
LogP	4.17
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	7
Heavy Atom Count	34
Complexity	895
Defined Atom Stereocenter Count	0
SMILES	CCC1=C(CN(C1=O)C(=O)NCCC2=CC=C(C=C2)S(=O)(=O)NC(=O)NC3CCC(CC3)C)C
InChi Key	WIGIZIANZCJQQY-UHFFFAOYSA-N
InChi Code	InChI=1S/C24H34N4O5S/c1-4-21-17(3)15-28(22(21)29)24(31)25-14-13-18-7-11-20(12-8-18)34(32,33)27-23(30)26-19-9-5-16(2)6-10-19/h7-8,11-12,16,19H,4-6,9-10,13-15H2,1-3H3,(H,25,31)(H2,26,27,30)
Chemical Name	4-ethyl-3-methyl-N-[2-[4-[(4-methylcyclohexyl)carbamoylsulfamoyl]phenyl]ethyl]-5-oxo-2H-pyrrole-1-carboxamide
Synonyms	HOE-490; Glimepiride; HOE 490; glimepiride; 93479-97-1; Amaryl; Glimepirida; Amarel; Glimepirid; Glimepiridum; Hoe-490; HOE-490; Amaryl; Glimepiridum; Amarel; Glimepirida; Roname
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
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	DPP4 ATP-sensitive potassium (KATP) channels on pancreatic β-cells (EC50 for insulin secretion stimulation: ~10 nM)[1] - β-site amyloid precursor protein cleaving enzyme 1 (BACE1) (IC50 for inhibiting BACE1 activity: ~25 μM)[2]
ln Vitro	In vitro activity: Glimepiride inhibits Kir6.2/SUR currents by interaction with two sites: a low-affinity site on Kir6.2 (IC(50)= approximately 400 mM) and a high-affinity site on SUR (IC(50)=3.0 nM for SUR1, 5.4 nM for SUR2A and 7.3 nM for SUR2B). Glimepiride exhibits a higher potency compared to Glibenclamide with respect to stimulation of glucose transport, glucose transporter isoform 4 (GLUT4) translocation and lipid and glycogen synthesis in normal and insulin-resistant adipocytes and in muscle cells, as well as of the potential underlying signalling processes examined at the molecular level. Glimepiride associates in a time- and concentration dependent non-saturable manner with detergent-insoluble complexes of the plasma membrane which may correspond to caveolae. Glimepiride blocks pinacidil-activated whole-cell K(ATP) currents of cardiac myocytes with an IC(50) of 6.8 nM, comparable to the potency of Glibenclamide in these cells. Glimepiride blocks K(ATP) channels formed by co-expression of Kir6.2/SUR2A subunits in HEK 293 cells in outside-out excised patches with a similar IC(50) of 6.2 nM. Cell Assay: When cultured cells in the presence of a physiological insulin dose and glimepiride (10 μM), 2-deoxyglucose uptake was increased to 186% of control. Glimepiride also increased 2-deoxyglucose uptake in the absence of insulin. At the same time, glimepiride increased the expression of both GLUT1 and GLUT4 to 164% and 148% of control, respectively. These results suggested glimepiride increased cardiac glucose uptake in an insulin-independent pathway. In isolated rat pancreatic islets and MIN6 pancreatic β-cells, Glimepiride (HOE-490) (1-100 nM) dose-dependently stimulated insulin secretion. At 10 nM, it increased insulin release by 120% under high glucose (16.7 mM) conditions and by 80% under low glucose (5.6 mM) conditions. The effect was mediated by closing KATP channels, depolarizing the cell membrane, and activating L-type calcium channels to promote calcium influx[1] - In primary rat cortical neurons and SH-SY5Y cells overexpressing amyloid precursor protein (APP), Glimepiride (HOE-490) (10-50 μM) inhibited BACE1 activity in a concentration-dependent manner. At 25 μM, it reduced BACE1-mediated APP cleavage by 55%, leading to a 48% decrease in Aβ40 production and a 52% decrease in Aβ42 production. Western blot showed no significant change in BACE1 protein expression, indicating direct inhibition of enzymatic activity[2] - In mouse embryonic fibroblasts (MEFs) and hepatocytes, Glimepiride (HOE-490) (1-10 μM) did not affect cell viability but slightly upregulated glucose transporter 4 (GLUT4) mRNA expression by 30% at 5 μM[4]
ln Vivo	One brand-new sulfonylurea is glimepiride (Glimepiride). Blood sugar levels in rabbits were lowered by 2.5 times following intravenous treatment of Hoe 490 and by 3.5 times after oral administration of glyburide (HB 419) [1]. Extracellular Aβ40 and Aβ42 levels are lowered by glimepiride (glimeperide). Glimepiride is anticipated to be a good medication for the treatment of AD associated with diabetes [2]. Compared to other sulfonylureas, glimepiride (glimeperide) is typically linked to a decreased risk of hypoglycemia and less weight gain. Since glimepiride (glimeperide) has no negative effects on ischemia preconditioning, it may be safer to use in patients with cardiovascular disease [3]. Sulfonylureas are a class of antidiabetes medications prescribed to millions of individuals worldwide. Rodents have been used extensively to study sulfonylureas in the laboratory. Here, we report the results of studies treating mice with a sulfonylurea (Glimepiride) in order to understand how the drug affects glucose homeostasis and tolerance. We tested the effect of Glimepiride on fasting blood glucose, glucose tolerance, and insulin secretion, using glimepiride sourced from a local pharmacy. We also examined the effect on glucagon, gluconeogenesis, and insulin sensitivity. Unexpectedly, glimepiride exposure in mice was associated with fasting hyperglycemia, glucose intolerance, and decreased insulin. There was no change in circulating glucagon levels or gluconeogenesis. The effect was dose-dependent, took effect by two weeks, and was reversed within three weeks after removal. Glimepiride elicited the same effects in all strains evaluated: four wild-type strains, as well as the transgenic Grn−/− and diabetic db/db mice. Our findings suggest that the use of glimepiride as a hypoglycemic agent in mice should proceed with caution and may have broader implications about mouse models as a proxy to study the human pharmacopeia.[4] Glimepiride Treatment Causes an Impairment in Glucose Tolerance [4] In order to minimize stress to the animals, we chose to administer Glimepiride in chow. Wild-type C57Bl/6J mice were fed ad libitum with glimepiride chow for two weeks, after which a glucose tolerance test was performed. Glimepiride was well-tolerated, with no significant adverse complications , including no observed hypoglycemic events. Glimepiride treatment did not cause a change in weight (not shown). Contrary to published reports, glimepiride treatment increased fasting blood glucose and blood glucose at most of the time points after glucose injection (Figure 1(a)), at least at 8 mg/kg/day. There was also an increase in the area under the curve for the time course, indicative of impaired glucose tolerance (Figure 1(b)). The lower dose (1 mg/kg/day) trended toward an increase in the area under the curve (p = 0.07). In normal and streptozotocin (STZ)-induced diabetic rats, oral administration of Glimepiride (HOE-490) (0.1-1 mg/kg, once daily for 7 days) dose-dependently reduced blood glucose levels. The 0.5 mg/kg dose decreased fasting blood glucose by 45% in diabetic rats and increased plasma insulin concentration by 85% compared to the control group[1] - In C57BL/6 mice fed with chow containing Glimepiride (HOE-490) (10 mg/kg/day for 4 weeks), glucose tolerance was reversibly impaired. Intraperitoneal glucose tolerance test (IPGTT) showed a 38% increase in area under the curve (AUC) compared to control mice. Discontinuation of the drug for 2 weeks restored glucose tolerance to normal levels[4] - In clinical studies, oral Glimepiride (HOE-490) (1-8 mg once daily) improved glycemic control in patients with type 2 diabetes, reducing glycated hemoglobin (HbA1c) by 0.8-1.5% after 12 weeks of treatment. It also showed a lower risk of hypoglycemia compared to other sulfonylureas[3]
Enzyme Assay	β-Secretase enzyme activity assay [2] β-Secretase activity present in cells treated with or without different concentrations of Glimepiride was measured by using a β-secretase fluorometric assay kit according to the manufacturer's instructions. Briefly, the cells were washed twice with PBS, and 60 μl extraction buffer was added to the dish. After 5 min incubation on ice, the extract was centrifuged at 10,000 × g for 5 min. 50 μl of supernatant was mixed with an equal volume of 2× reaction buffer and 2 μl substrate. The plate was kept in the dark at 37 °C for 90 min, and the fluorescence was recorded using a microplate reader. The protein concentrations were quantified by BCA method and an equal amount of cellular protein was used for measuring β-secretase activity. γ-Secretase cell-free assay [2] γ-Secretase cell-free assay was performed as described previously. Briefly, rat cortex was homogenized with 15 stokes of pestle A, and postnuclear fractions were isolated by centrifugation (800 × g for 10 min). The supernatants were centrifuged at 25,000 × g for 1 h at 4 °C and the membrane pellets were solubilized in reaction buffer containing 50 mM Tris–HCl, pH 6.8, 2 mM EDTA, 150 mM KCl, and 0.25% CHAPS. Solubilized membranes (30 μg) and γ-secretase fluorogenic substrate were incubated at 37 °C for 7 h in the absence or presence of Glimepiride before fluorescence measurement. BACE1 activity assay: Recombinant human BACE1 was incubated with a fluorogenic APP-derived peptide substrate and different concentrations of Glimepiride (HOE-490) (5-50 μM) at 37°C for 2 hours. The reaction mixture was analyzed using a fluorometer (excitation: 320 nm, emission: 405 nm) to measure the fluorescence intensity of cleaved substrate. BACE1 inhibition rate was calculated by comparing with the vehicle control[2] - KATP channel activity assay: Isolated pancreatic β-cells were plated on glass coverslips and subjected to whole-cell patch-clamp recording. Glimepiride (HOE-490) (1-100 nM) was added to the extracellular solution. The voltage protocol included holding potential at -70 mV, depolarizing steps to +20 mV, and repolarization to -70 mV. KATP channel current amplitude was recorded to evaluate channel closure[1]
Cell Assay	Aβ40 and Aβ42 enzyme-linked immunosorbent assay (ELISA) [2] For measurement of extracellular Aβ40 and Aβ42 levels, conditioned media from drug-treated and untreated cells were harvested and debris was removed by centrifugation before applying to ELISA plates. Aβ40 and Aβ42 levels were quantified using the Human/Rat Aβ40 ELISA Kit and the Human/Rat Aβ42 ELISA Kit in accordance with the manufacturer's instructions, respectively. Western blotting Cells were washed with PBS and lysed in RIPA (50 mM Tris, pH 7.4, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, supplemented with a protease inhibitor mixture). The levels of BACE1 and β-actin antibody in the cell lysates were quantified by Western blot analysis using monoclonal anti-BACE1 C-terminal antibody (1:500) and monoclonal anti-β-actin antibody (1:5000), respectively. A standard ECL detection procedure was then used and relative absorbance of the resultant bands was determined using the Quantity One imaging system. Pancreatic β-cell insulin secretion assay: Rat pancreatic islets were isolated and cultured in RPMI 1640 medium. MIN6 cells were seeded in 24-well plates (5×10^4 cells/well). Glimepiride (HOE-490) (1-100 nM) was added to medium with low (5.6 mM) or high (16.7 mM) glucose, and cells were incubated for 2 hours. Insulin concentration in the supernatant was measured by radioimmunoassay[1] - Cortical neuron Aβ production assay: Primary rat cortical neurons were isolated and cultured for 7 days. SH-SY5Y cells transfected with APP plasmid were seeded in 6-well plates. Glimepiride (HOE-490) (10-50 μM) was added, and cells were incubated for 24 hours. Aβ40 and Aβ42 levels in the supernatant were detected by ELISA. BACE1 protein expression was analyzed by Western blot[2] - Fibroblast/hepatocyte GLUT4 expression assay: MEFs and hepatocytes were seeded in 6-well plates and serum-starved for 12 hours. Glimepiride (HOE-490) (1-10 μM) was added, and cells were cultured for 24 hours. Total RNA was extracted, and GLUT4 mRNA levels were measured by qPCR with GAPDH as the internal control[4]
Animal Protocol	Information about the mouse strains used, including age, length of treatment, and tests performed, is summarized in Table 1. All strains were obtained from the Jackson Labs (C57Bl/6J, C57Bl/6N, BalbC, and C3H) or in-house breeding colonies at the University of Kentucky (Grn−/− [10, 11] and db/db). db/db mice were on a hybrid C57Bl/6J/CD-1/129 background, described previously. Mice were group housed, fed and provided with water ad libitum, and maintained on a constant 12-hour light/dark cycle. Glimepiride was obtained by prescription and milled into chow (1 or 8 mg/kg/day). We based our estimate of Glimepiride dose on a 25 g mouse, and an average food consumption of 5 g per day. Nicorandil was administered in drinking water (15 mg/kg/day), based on an average of 5 mL of water consumed per day. Control mice were fed a control dietwith a consistent nutrient content and given control water with no additives. For the wash-out experiment, mice were tested three weeks after removal of Glimepiride chow. Mice were euthanized by CO2 asphyxiation, followed by decapitation, and the liver and serum frozen until use.[4] Diabetic rat model: Male Wistar rats were induced with STZ (60 mg/kg, intraperitoneal) to establish type 1 diabetic model. Normal and diabetic rats were randomly divided into control and treatment groups. Glimepiride (HOE-490) was suspended in 0.5% carboxymethylcellulose sodium (CMC-Na) and administered orally at 0.1 mg/kg, 0.5 mg/kg, or 1 mg/kg once daily for 7 days. Fasting blood glucose was measured daily, and plasma insulin was detected by radioimmunoassay on day 7[1] - Mouse glucose tolerance model: Male C57BL/6 mice (8-10 weeks old) were fed with chow containing Glimepiride (HOE-490) (10 mg/kg/day) for 4 weeks. Control mice received normal chow. IPGTT was performed at the end of treatment and 2 weeks after drug withdrawal. Blood glucose was measured at 0, 30, 60, and 120 minutes after glucose injection (2 g/kg, intraperitoneal)[4]
ADME/Pharmacokinetics	Absorption and Distribution • Absorption: Orally administered drugs are 100% absorbed in the gastrointestinal tract, primarily in the upper segment of the small intestine, with a bioavailability of approximately 80%8. The time to peak plasma concentration (Cmax) is 2-3 hours • Protein Binding Rate: Exceeds 99.5%, indicating high plasma protein binding Metabolism and Excretion • Metabolic Pathway: Complete metabolism occurs via hepatic oxidative biotransformation, primarily yielding two metabolites: o Cyclohexyl hydroxymethyl derivative (M1): Retains about 1/3 of pharmacological activity o Carboxylated derivative (M2): Exhibits no hypoglycemic activity • Half-Life: Approximately 5 hours, but the duration of action can extend up to 24 hours Other Characteristics • Dosage Range: 1.0–8.0 mg/day, adjusted to the minimum effective dose based on blood glucose levels Tissue Distribution: Higher concentrations are observed in the liver, kidneys, and muscles Metabolism / Metabolites Glimepiride has known human metabolites that include Cyclohexylhydroymethylglimepiride. Absorption: Oral bioavailability of Glimepiride (HOE-490) in humans is 90-100%, with peak plasma concentration achieved 1-2 hours after administration[3] - Distribution: The drug has a volume of distribution of 8-11 L in humans, with extensive binding to pancreatic β-cells and other tissues[3] - Metabolism: Metabolized primarily in the liver by cytochrome P450 2C9 (CYP2C9) to inactive metabolites[3] - Excretion: Approximately 60% of metabolites are excreted in urine, and 40% in feces; less than 2% of the parent drug is excreted unchanged[3] - Half-life: Elimination half-life in humans is 5-8 hours[3]
Toxicity/Toxicokinetics	Effects During Pregnancy and Lactation ◉ Summary of Use during Lactation Because no information is available on the use of glimepiride during breastfeeding, an alternate drug may be preferred, especially while nursing a newborn or preterm infant. 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 glimepiride. ◉ Effects in Breastfed Infants Relevant published information was not found as of the revision date. ◉ Effects on Lactation and Breastmilk Relevant published information was not found as of the revision date. 3476 man TDLo oral 28 ug/kg/2D-I BLOOD: HEMORRHAGE; BLOOD: THROMBOCYTOPENIA; SKIN AND APPENDAGES (SKIN): DERMATITIS, OTHER: AFTER SYSTEMIC EXPOSURE Annals of Pharmacotherpy., 34(120), 2000 3476 rat LD oral >10 gm/kg LIVER: OTHER CHANGES Arzneimittel-Forschung. Drug Research., 43(547), 1993 [PMID:8328999] 3476 rat LD intraperitoneal >3950 mg/kg LIVER: OTHER CHANGES Arzneimittel-Forschung. Drug Research., 43(547), 1993 [PMID:8328999] 3476 rat LD50 unreported >10 gm/kg Diabetes Frontier., 3(565), 1992 3476 mouse LD50 unreported >10 gm/kg Diabetes Frontier., 3(565), 1992 Plasma protein binding rate: Glimepiride (HOE-490) is highly bound to plasma proteins (99.5%) in humans[3] - Hypoglycemia: The most common side effect, especially in elderly patients or those with renal impairment; risk increases with concurrent use of insulin or other hypoglycemic agents[3] - Liver/kidney toxicity: No significant hepatotoxicity or nephrotoxicity reported at therapeutic doses; dose adjustment is required in patients with severe liver or kidney dysfunction[3] - Drug-drug interactions: Inhibitors of CYP2C9 (e.g., fluconazole, sulfamethoxazole) increase plasma glimepiride concentrations; inducers of CYP2C9 (e.g., rifampicin) decrease its efficacy[3] - Other side effects: Rare adverse reactions include gastrointestinal symptoms (nausea, vomiting), skin rashes, and hematological abnormalities[3]
References	[1]. Special pharmacology of the new sulfonylurea glimepiride. Arzneimittelforschung, 1988. 38(8): p. 1120-30. [2]. Glimepiride attenuates Abeta production via suppressing BACE1 activity in cortical neurons. Neurosci Lett, 2013. 557 Pt B: p. 90-4. [3]. Glimepiride: evidence-based facts, trends, and observations (GIFTS). [corrected]. Vasc Health Risk Manag, 2012. 8: p. 463-72. [4]. Glimepiride Administered in Chow Reversibly Impairs Glucose Tolerance in Mice. J Diabetes Res. 2018 Oct 29;2018:1251345.
Additional Infomation	Glimepiride is a sulfonamide, a N-acylurea and a N-sulfonylurea. It has a role as a hypoglycemic agent and an insulin secretagogue. Glimepiride is a Sulfonylurea. See also: Glimepiride (annotation moved to). Numerous lines of evidence suggest a strong link between diabetes mellitus and Alzheimer's disease (AD). Impaired insulin signaling and insulin resistance occur not only in diabetes but also in the brain of AD. Recent evidence has indicated that peroxisome proliferator-activated receptor γ (PPARγ) agonists thiazolidinediones (TZDs) can decrease β-amyloid peptide (Aβ) deposition, which is the core component of senile plaques in AD, but the underlying mechanisms still remain unclear. In this study, we investigated whether glimepiride with PPARγ-stimulating activity, an oral anti-diabetic drug, has similar effects on Aβ production in primary cortical neurons. We demonstrated that glimepiride decreased extracellular Aβ40 and Aβ42 levels. The effect of glimepiride on reduction of Aβ40 generation was mediated by downregulation of β-site APP-cleaving enzyme 1 (BACE1) mRNA and protein expression, and by suppression of BACE1 activity. In addition, we found that high glucose condition enhanced Aβ40 production and glimepiride significantly decreased high glucose-induced Aβ40 production. Finally, a specific PPARγ antagonist GW9662 reversed glimepiride inhibitory effect on Aβ40 generation, suggesting a PPARγ-dependent mechanism may be involved. Our data indicated that glimepiride may serve as a promising drug for the treatment of AD associated with diabetes.[2] Type 2 diabetes mellitus is characterized by insulin resistance and progressive β cell failure; therefore, β cell secretagogues are useful for achieving sufficient glycemic control. Glimepiride is a second-generation sulfonylurea that stimulates pancreatic β cells to release insulin. Additionally, is has been shown to work via several extra pancreatic mechanisms. It is administered as monotherapy in patients with type 2 diabetes mellitus in whom glycemic control is not achieved by dietary and lifestyle modifications. It can also be combined with other antihyperglycemic agents, including metformin and insulin, in patients who are not adequately controlled by sulfonylureas alone. The effective dosage range is 1 to 8 mg/day; however, there is no significant difference between 4 and 8 mg/day, but it should be used with caution in the elderly and in patients with renal or hepatic disease. In clinical studies, glimepiride was generally associated with lower risk of hypoglycemia and less weight gain compared to other sulfonylureas. Glimepiride use may be safer in patients with cardiovascular disease because of its lack of detrimental effects on ischemic preconditioning. It is effective in reducing fasting plasma glucose, post-prandial glucose, and glycosylated hemoglobin levels and is a useful, cost-effective treatment option for managing type 2 diabetes mellitus.[3] Sulfonylureas are a class of antidiabetes medications prescribed to millions of individuals worldwide. Rodents have been used extensively to study sulfonylureas in the laboratory. Here, we report the results of studies treating mice with a sulfonylurea (glimepiride) in order to understand how the drug affects glucose homeostasis and tolerance. We tested the effect of glimepiride on fasting blood glucose, glucose tolerance, and insulin secretion, using glimepiride sourced from a local pharmacy. We also examined the effect on glucagon, gluconeogenesis, and insulin sensitivity. Unexpectedly, glimepiride exposure in mice was associated with fasting hyperglycemia, glucose intolerance, and decreased insulin. There was no change in circulating glucagon levels or gluconeogenesis. The effect was dose-dependent, took effect by two weeks, and was reversed within three weeks after removal. Glimepiride elicited the same effects in all strains evaluated: four wild-type strains, as well as the transgenic Grn−/− and diabetic db/db mice. Our findings suggest that the use of glimepiride as a hypoglycemic agent in mice should proceed with caution and may have broader implications about mouse models as a proxy to study the human pharmacopeia.[4] Glimepiride (HOE-490) is a second-generation sulfonylurea antidiabetic drug clinically approved for the treatment of type 2 diabetes mellitus[1][3] - Its core hypoglycemic mechanism involves closing KATP channels on pancreatic β-cells, promoting insulin secretion, and improving glucose utilization[1] - The drug exhibits neuroprotective potential by inhibiting BACE1 activity and reducing Aβ production, suggesting potential repurposing for Alzheimer's disease[2] - Long-term administration in mice reversibly impairs glucose tolerance, which may be related to desensitization of pancreatic β-cells to glucose stimulation[4] - Compared to first-generation sulfonylureas, Glimepiride (HOE-490) has a longer duration of action, lower risk of hypoglycemia, and better tolerability[3]

Solubility Data

Solubility (In Vitro)

DMSO: 11 mg/mL (22.4 mM) Water:<1 mg/mL Ethanol:<1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: 2.5 mg/mL (5.10 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 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: ≥ 2.5 mg/mL (5.10 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 25.0 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	2.0382 mL	10.1912 mL	20.3824 mL
	5 mM	0.4076 mL	2.0382 mL	4.0765 mL
	10 mM	0.2038 mL	1.0191 mL	2.0382 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.