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]

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.

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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.

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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]

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].

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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]

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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).

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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]

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.

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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.

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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]

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.

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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.

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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]

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]

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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]

Absorption and Distribution
• Absorption: Orally administered drugs are 100% absorbed in the gastrointestinal tract, mainly in the upper small intestine, with a bioavailability of approximately 80%8. Time to peak concentration (Cmax) is 2-3 hours.
• Protein binding: Over 99.5%, indicating high plasma protein binding.
Metabolism and excretion
• Metabolic pathway: Complete metabolism occurs via hepatic oxidative biotransformation, primarily producing two metabolites:
o Cyclohexylhydroxymethyl derivative (M1): Retains approximately 1/3 of the pharmacological activity.
o Carboxylated derivative (M2): Does not have hypoglycemic activity.
• Half-life: Approximately 5 hours, but the duration of action can be up to 24 hours.
Other characteristics
• Dosage range: 1.0–8.0 mg/day, adjusted to the lowest effective dose based on blood glucose levels.
Tissue distribution: High concentrations are observed in the liver, kidneys, and muscle.
Metabolism/Metabolites
Glimepiride's known metabolites include cyclohexylhydroxymethyl glimepiride.

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Absorption: The oral bioavailability of glimepiride (HOE-490) in the human body is 90-100%, and peak plasma concentration is reached 1-2 hours after administration [3]
-Distribution: The volume of distribution of this drug in the human body is 8-11 liters, and it binds extensively to pancreatic β cells and other tissues [3]
-Metabolism: It is mainly metabolized in the liver by cytochrome P450 2C9 (CYP2C9) into inactive metabolites [3]
-Excretion: About 60% of the metabolites are excreted in urine and 40% in feces; less than 2% of the parent drug is excreted unchanged [3]
-Half-life: The elimination half-life in the human body is 5-8 hours [3]

Effects During Pregnancy and Lactation
◉ Overview of Use During Lactation
Since there is currently no information regarding the use of glimepiride during lactation, it is recommended to prioritize other medications, especially when breastfeeding newborns or premature infants. Monitor breastfed infants for signs of hypoglycemia, such as irritability, lethargy, feeding difficulties, seizures, cyanosis, apnea, or hypothermia. If there is any concern, it is recommended to monitor the breastfed infant's blood glucose levels while the mother is taking glimepiride.
◉ Effects on Breastfed Infants
As of the revision date, no relevant published information was found.
◉ Effects on Lactation and Breast Milk
As of the revision date, no relevant published information was found.

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3476 human TDLo oral 28 ug/kg/2D-I Blood: hemorrhage; Blood: thrombocytopenia; Skin and appendages (skin): dermatitis, other: post-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 injection >3950 mg/kg Liver: other changes Arzneimittel-Forschung. Drug Research, 43(547), 1993 [PMID:8328999]
3476 rat LD50 not reported >10 gm/kg Frontiers in Diabetes, 3(565), 1992
3476 Mouse LD50 Not reported >10 gm/kg Frontiers in Diabetes, 3(565), 1992
Plasma protein binding:Glimepiride (HOE-490) is highly bound to plasma proteins in humans (99.5%)[3]
- Hypoglycemia: The most common side effect, especially in elderly patients or patients with renal insufficiency; the risk is increased with the use of insulin or other hypoglycemic agents[3]
- Hepatotoxicity/nephrotoxicity: No significant hepatotoxicity or nephrotoxicity has been reported at therapeutic doses; dose adjustment is required in patients with severe hepatotoxicity or renal insufficiency[3]
- Drug interactions: CYP2C9 inhibitors (e.g., fluconazole, sulfamethoxazole) can increase plasma concentrations of glimepiride; CYP2C9 inducers (e.g., rifampin) can reduce its efficacy[3]
- Other side effects: Rare adverse reactions include gastrointestinal symptoms (nausea, vomiting), rash, and hematologic abnormalities [3]

[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.

Glimepiride is a sulfonamide drug belonging to the N-acylurea and N-sulfonylurea classes. It has hypoglycemic and insulin-secreting effects. Glimepiride belongs to the sulfonylurea class. See also: Glimepiride (note moved to). Extensive evidence suggests a close link between diabetes and Alzheimer's disease (AD). Impaired insulin signaling and insulin resistance are not only seen in diabetes but also in the brains of AD patients. Recent evidence suggests that peroxisome proliferator-activated receptor gamma (PPARγ) agonists thiazolidinediones (TZDs) can reduce the deposition of β-amyloid (Aβ), a core component of senile plaques in AD, but the underlying mechanism remains unclear. This study investigated whether the oral hypoglycemic agent glimepiride (with PPARγ stimulating activity) has a similar effect on Aβ production in primary cortical neurons. The results showed that glimepiride reduced extracellular Aβ40 and Aβ42 levels. The mechanism by which glimepiride reduces Aβ40 production is by downregulating the mRNA and protein expression of β-site APP lyase 1 (BACE1) and inhibiting BACE1 activity. In addition, we found that high glucose conditions enhance Aβ40 production, while glimepiride significantly reduces high glucose-induced Aβ40 production. Finally, the specific PPARγ antagonist GW9662 reversed the inhibitory effect of glimepiride on Aβ40 production, suggesting a possible PPARγ-dependent mechanism. Our data suggest that glimepiride may be a promising drug for the treatment of diabetes-related Alzheimer's disease (AD). [2] Type 2 diabetes is characterized by insulin resistance and progressive β-cell dysfunction; therefore, β-cell secretagogues help achieve adequate glycemic control. Glimepiride is a second-generation sulfonylurea that stimulates the release of insulin from pancreatic β-cells. In addition, studies have shown that it can also act through a variety of extrapancreatic mechanisms. Glimepiride can be used as monotherapy for patients with type 2 diabetes whose glycemic control is not achieved through diet and lifestyle modifications. For patients whose blood glucose is poorly controlled by sulfonylureas alone, glimepiride can also be used in combination with other hypoglycemic agents, including metformin and insulin. The effective dose range is 1–8 mg/day; however, there is no significant difference between 4 mg/day and 8 mg/day, but caution should be exercised in elderly patients and patients with kidney or liver disease. In clinical studies, glimepiride is generally associated with a lower risk of hypoglycemia and less weight gain compared to other sulfonylureas. Since glimepiride has no adverse effects on ischemic preconditioning, it may be safer to use in patients with cardiovascular disease. It effectively lowers fasting blood glucose, postprandial blood glucose and glycated hemoglobin levels, making it an effective and economical treatment option for type 2 diabetes. [3] Sulfonylureas are a class of antidiabetic drugs that are taken by millions of patients worldwide. Rodents have been widely used in laboratory studies of sulfonylureas. This article reports the results of a study in mice treated with sulfonylureas (glimepiride) to understand how the drug affects glucose homeostasis and glucose tolerance. We used glimepiride, obtained from a local pharmacy, to test its effects on fasting blood glucose, glucose tolerance, and insulin secretion. We also investigated the effects of glimepiride on glucagon, gluconeogenesis, and insulin sensitivity. Unexpectedly, mice administered glimepiride developed fasting hyperglycemia, glucose intolerance, and decreased insulin levels. Circulating glucagon levels and gluconeogenesis remained unchanged. This effect was dose-dependent, appearing within two weeks and reversible within three weeks of discontinuation. Glimepiride produced the same effects in all mouse strains evaluated: including four wild-type mouse strains, as well as transgenic Grn−/− mice and diabetic db/db mice. Our results suggest that the use of glimepiride as a hypoglycemic agent in mice should be approached with caution and may have broader implications for studying human drugs using mouse models. [4]

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Glimepiride (HOE-490) is a second-generation sulfonylurea antidiabetic drug that has been clinically approved for the treatment of type 2 diabetes[1][3]
- Its core hypoglycemic mechanism includes closing KATP channels on pancreatic β cells, promoting insulin secretion, and improving glucose utilization[1]
- The drug has neuroprotective effects by inhibiting BACE1 activity and reducing Aβ production, suggesting that it may be used to treat Alzheimer's disease[2]
- Long-term use of the drug in mice reversibly impairs glucose tolerance, which may be related to the desensitization of pancreatic β cells to glucose stimulation[4]
- Compared with first-generation sulfonylureas, glimepiride (HOE-490) has a longer duration of action, a lower risk of hypoglycemia, and better tolerability[3]

C24H34N4O5S

490.62

490.224

C, 58.75; H, 6.99; N, 11.42; O, 16.31; S, 6.54

93479-97-1

Glimepiride-d5;1028809-90-6; Glimepiride-d4-1; 1131981-29-7; 119018-30-3 (urethane); 119018-29-0 (sulfonamide); 93479-97-1

3476

White to off-white solid powder

1.3±0.1 g/cm3

677.0±65.0 °C at 760 mmHg

212.2-214.5 °C

363.2±34.3 °C

0.0±2.2 mmHg at 25°C

1.628

4.17

3

5

7

34

895

0

CCC1=C(CN(C1=O)C(=O)NCCC2=CC=C(C=C2)S(=O)(=O)NC(=O)NC3CCC(CC3)C)C

WIGIZIANZCJQQY-UHFFFAOYSA-N

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)

4-ethyl-3-methyl-N-[2-[4-[(4-methylcyclohexyl)carbamoylsulfamoyl]phenyl]ethyl]-5-oxo-2H-pyrrole-1-carboxamide

HOE-490; Glimepiride; HOE 490; glimepiride; 93479-97-1; Amaryl; Glimepirida; Amarel; Glimepirid; Glimepiridum; Hoe-490; HOE-490; Amaryl; Glimepiridum; Amarel; Glimepirida; Roname

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 (5.10 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.

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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.

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