Gastric H+/K+ ATPase proton pump. The study demonstrates that esomeprazole‘s effects on triple-negative breast cancer cells are mediated through this target, evidenced by increased expression of the pump in sensitive cells. [1]
The study suggests that esomeprazole, beyond its classical target gastric H+/K+ ATPase, can inhibit dimethylarginine dimethylaminohydrolase (DDAH) and downregulate inducible nitric oxide synthase (iNOS) expression and activity, contributing to its anti-inflammatory and anti-fibrotic effects. [2]

Esomeprazole (25-100 µM; 20 hours; MDA-MB-468 cells) treatment enhances intracellular acidification, which in turn inhibits the growth of triple-negative breast cancer cells in vitro in a dose-dependent manner [1].

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Esomeprazole suppressed the growth of triple-negative breast cancer cell line MDA-MB-468 in a dose-dependent manner with an EC50 of approximately 70 µM, as determined by trypan blue exclusion assay after 20 hours of treatment. [1]
Esomeprazole (at 50 µM) significantly increased intracellular acidity in MDA-MB-468 cells, reducing the relative fluorescence of the pH-sensitive probe BCECF-AM to 60.2% of control levels. This acidification is linked to its growth-suppressive mechanism. [1]
Esomeprazole enhanced the cytotoxic effects of doxorubicin (DOX) on MDA-MB-468 cells. Combination treatment with fixed concentrations of esomeprazole (30 µM) and doxorubicin (30 nM) resulted in a significantly greater reduction in live cell number compared to doxorubicin treatment alone. [1]
In contrast to cancer cells, the non-cancerous breast epithelial cell line MCF-10A was significantly less sensitive to esomeprazole. Treatment with 100 µM esomeprazole reduced the number of live MDA-MB-468 cells to 51.88% of control, while the reduction in MCF-10A cells (to 92.31% of control) was not statistically significant. [1]

The C57BL/6J mice treated with esomeprazole (30–300 mg/kg; oral gavage; daily; for 19 or 11 days) showed a significant reduction in the animals' lung fibrosis progression. Additionally, esomeprazole lowers circulating fibrosis and inflammatory markers [2].

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In a mouse model of cotton smoke-induced lung injury over 3 weeks, therapeutic administration of a low dose of esomeprazole (30 mg/kg/day, p.o., starting 10 days post-smoke exposure) significantly increased plasma levels of asymmetric dimethylarginine (ADMA) and reduced plasma nitric oxide (NO) levels compared to vehicle-treated controls, indicating modulation of the DDAH/iNOS pathway. [2]
The same therapeutic dose (30 mg/kg/day) significantly reduced circulating levels of pro-inflammatory and pro-fibrotic markers, including tumor necrosis factor-alpha (TNFα) and matrix metalloproteinase-7 (MMP7), compared to vehicle-treated controls exposed to smoke. A decreasing trend was also observed for interleukin-1β (IL1β). [2]
Therapeutic administration of low-dose esomeprazole (30 mg/kg/day) significantly inhibited the progression of lung fibrosis. The mean fibrosis score in the treatment group (0.64) was significantly lower than that in the vehicle control group (1.06). Histological analysis showed minimal to no fibrosis in the treatment group, whereas the control group exhibited fibrous thickening of alveolar walls. [2]
Treatment with esomeprazole did not show a significant favorable effect on lung inflammation (assessed by H&E staining for inflammatory cell infiltration) in this specific smoke-induced injury model. [2]
A high prophylactic dose of esomeprazole (300 mg/kg/day, p.o., starting 2 days post-smoke exposure) was poorly tolerated, leading to increased mortality, bloating, and sluggishness in mice, and showed no meaningful impact on suppressing lung fibrosis. [2]

Cell Viability Assay[1]
Cell Types: MDA-MB-468 Cell
Tested Concentrations: 25 µM, 50 µM, 75 µM, 100 µM
Incubation Duration: 20 hrs (hours)
Experimental Results: Inhibition of triple negative breast cancer cells in a dose-dependent manner in vitro.

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Cell Viability/Growth Suppression Assay: Triple-negative breast cancer MDA-MB-468 cells and non-cancerous breast epithelial MCF-10A cells were cultured in their respective growth media. Cells were treated with varying concentrations of esomeprazole, doxorubicin, or their combination in 6-well plates for 20 hours in RPMI medium (chosen for its low buffering capacity) containing fetal bovine serum. After treatment, cells were washed, detached with trypsin, and stained with trypan blue. The number of live (unstained) cells was counted using an automated cell counter. Data were expressed relative to untreated control cells. [1]
Intracellular pH Measurement: MDA-MB-468 cells were treated with esomeprazole (50 µM) or acidified medium (pH 6.0) for 20 hours. After treatment, cells were washed and incubated with 3 µM BCECF-AM, a pH-sensitive fluorescent probe, for 30 minutes at 37°C. Following another wash, intracellular fluorescence was detected and imaged using confocal microscopy. The fluorescence intensity at 530 nm, which is inversely correlated with intracellular acidity (lower pH = lower fluorescence), was quantified via digital image analysis and expressed as relative fluorescence per cell. [1]
Immunofluorescence for Target Detection: MDA-MB-468 and MCF-10A cells were grown on glass coverslips. Cells were fixed with paraformaldehyde, permeabilized, and incubated with a primary mouse monoclonal antibody against the human H+/K+ ATPase β subunit. After washing, cells were incubated with a fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG secondary antibody. Fluorescence was visualized and detected using confocal microscopy to assess the expression level of the gastric proton pump. [1]
Western Blotting for Target Confirmation: Cell lysates from MDA-MB-468 and MCF-10A cells were prepared. Proteins were separated by electrophoresis, transferred to a membrane, and probed with the same primary mouse monoclonal antibody against human H+/K+ ATPase β subunit used in immunofluorescence. A horseradish peroxidase-conjugated goat anti-mouse IgG was used as the secondary antibody for detection. To confirm equal protein loading, the membrane was also probed with a mouse monoclonal antibody against human α-tubulin. [1]

Animal/Disease Models: C57BL/6J mice (8 weeks old, 25-30 g) cotton smoke-induced lung injury [2]
Doses: 30 mg/kg, 300 mg/kg
Route of Administration: po (oral gavage); daily; continued for 19 Or 11-day
Experimental Results: Dramatically inhibited the progression of lung fibrosis in animals.

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Cotton Smoke-Induced Lung Injury Model in Mice: Eight-week-old C57BL/6J mice were exposed to cotton smoke for 21 days to induce lung injury. Animals were randomized into groups: a no-exposure (sham) group, a smoke-exposed vehicle control group, a prophylactic esomeprazole group, and a therapeutic esomeprazole group. The vehicle and prophylactic groups received daily oral gavage of either 10% ethanol (vehicle) or esomeprazole (300 mg/kg dissolved in 10% ethanol) starting 2 days after the initiation of smoke exposure. The therapeutic group received daily oral esomeprazole at a lower dose of 30 mg/kg (in 10% ethanol) starting 10 days after smoke exposure began and continuing until the end of the study (necropsy). At necropsy, blood was collected via cardiac puncture for plasma analysis, and lungs and other organs were harvested for weight measurement and histopathological examination (H&E and Masson's Trichrome staining). [2]

Absorption, Distribution and Excretion
Following oral administration, peak plasma concentration (Cmax) is reached at approximately 1.5 hours (Tmax). Cmax increases proportionally with dose, and the area under the plasma concentration-time curve (AUC) triples from 20 mg to 40 mg. Systemic bioavailability is approximately 90% with repeated once-daily administration of 40 mg, compared to approximately 64% with a single 40 mg dose. Following once-daily administration of 40 mg, the mean exposure (AUC) of esomeprazole increases from 4.32 μmolhr/L on day 1 to 11.2 μmolhr/L on day 5. The AUC decreases by 43% to 53% after a single 40 mg dose of esomeprazole taken after a meal compared to a fasting state. Esomeprazole should be taken at least one hour before a meal. _Combined Antibiotic Therapy:_ Seventeen healthy male and female subjects received 40 mg esomeprazole magnesium once daily, combined with 500 mg twice daily [DB01211] and 1000 mg twice daily [DB01060] for 7 days. Compared with esomeprazole monotherapy, the mean steady-state AUC and Cmax of esomeprazole increased by 70% and 18%, respectively, during triple therapy. The increased esomeprazole exposure observed during concomitant use with clarithromycin and amoxicillin is not expected to cause significant safety issues.
The plasma elimination half-life of esomeprazole is approximately 1 to 1.5 hours. Less than 1% of the unchanged drug is excreted in the urine. After oral administration of esomeprazole, approximately 80% is excreted in the urine as inactive metabolites, and the remainder is excreted in the feces as inactive metabolites.
The apparent volume of distribution at steady state in healthy volunteers is approximately 16 liters.
The plasma elimination half-life of esomeprazole is approximately 1 to 1.5 hours. Less than 1% of the unchanged drug is excreted in the urine. After oral administration of esomeprazole, approximately 80% is excreted in the urine as inactive metabolites, and the remainder is excreted in the feces as inactive metabolites.
Esomeprazole binds to plasma proteins at a rate of 97%. Plasma protein binding remains constant within a concentration range of 2 to 20 μmol/L. In healthy volunteers, the steady-state apparent volume of distribution is approximately 16 liters.
Nexium extended-release capsules and Nexium extended-release oral suspension contain bioequivalent esomeprazole magnesium enteric-coated granules. Bioequivalence is based on a single-dose (40 mg) study conducted in 94 healthy male and female volunteers on an empty stomach. Peak plasma concentration (Cmax) occurs at approximately 1.5 hours (Tmax) after oral administration. Cmax increases proportionally with dose, and the area under the plasma concentration-time curve (AUC) triples with increasing dose from 20 mg to 40 mg. Systemic bioavailability is approximately 90% with repeated once-daily administration of 40 mg, compared to approximately 64% with a single 40 mg dose. The mean exposure (AUC) of esomeprazole increased from 4.32 μmol/hr/L on day 1 to 11.2 μmol/hr/L on day 5 after a once-daily administration of 40 mg.

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Metabolism/Metabolites
Esomeprazole is primarily metabolized in the liver via the cytochrome P450 (CYP) enzyme system. Esomeprazole metabolites do not possess antisecretory activity. The majority of esomeprazole metabolism depends on the CYP2C19 isoenzyme, producing hydroxyl and demethylated metabolites. The remainder depends on CYP3A4, producing sulfone metabolites. The CYP2C19 isoenzyme exhibits polymorphism in the metabolism of esomeprazole. Approximately 3% of Caucasians and 15% to 20% of Asians lack CYP2C19 and are termed astomosomal metabolizers. However, the effect of CYP2C19 polymorphism on esomeprazole is less significant than its effect on omeprazole. At steady state, the AUC ratio of astomosomal metabolizers to the AUC ratio of the remaining population (metabolic metabolizers) is approximately 2. After administration of equimolar doses, the S- and R-isomers are metabolized differently in the liver, resulting in higher plasma concentrations of the S-isomer than the R-isomer. Nine major urinary metabolites have been detected. Two of these major metabolites have been identified as hydroxyesomeprazole and its corresponding carboxylic acid. Three major metabolites have been identified in plasma: 5-O-demethyl derivatives, sulfone derivatives, and hydroxyesomeprazole. The major metabolites of esomeprazole have no effect on gastric acid secretion. Esomeprazole is primarily metabolized in the liver via the cytochrome P450 (CYP) enzyme system. Esomeprazole metabolites do not possess antisecretory activity. The majority of esomeprazole metabolism depends on the CYP 2C19 isoenzyme, which produces hydroxyl and demethylated metabolites. The remainder depends on CYP 3A4, which produces sulfone metabolites. CYP 2C19 isoenzymes exhibit polymorphism in esomeprazole metabolism, as approximately 3% of Caucasians and 15% to 20% of Asians lack CYP 2C19 and are termed as poor metabolizers. At steady state, the AUC ratio of slower metabolizers to that of the rest of the population (faster metabolizers) is approximately 2. Following equimolar dose administration, the S- and R-isomers are metabolized differently in the liver, resulting in higher plasma concentrations of the S-isomer than the R-isomer.

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Biological half-life
1–1.5 hours

Effects During Pregnancy and Lactation
◉ Overview of Use During Lactation
Esomeprazole is the S-enantiomer of the proton pump inhibitor omeprazole. Limited information suggests that a mother taking 10 mg of esomeprazole daily, with low concentrations in breast milk, is not expected to have any adverse effects on the breastfed infant.
◉ Effects on Breastfed Infants
One mother took 20 mg of omeprazole orally daily, expressing and discarding breast milk four hours after each morning dose. She then continued breastfeeding for three months until weaning. The infant was in good health at 12 months of age.
A woman with rheumatoid arthritis received oral esomeprazole 10 mg, prednisone 2.5 mg, and sulfasalazine 1 g once daily, along with injections of 200 mg of cetuzumab every two weeks. Her baby was approximately 50% breastfed and 50% formula-fed. No detectable drug-related adverse reactions were observed in the infant.
◉ Effects on Breastfeeding and Lactation
Omeprazole (racemic mixture) has been reported to cause gynecomastia in several men. A retrospective US claims database study found an increased risk of gynecomastia in users of proton pump inhibitors.
A review article reported that a search of the European Pharmacovigilance Center database found 45 cases of gynecomastia, 9 cases of galactorrhea, 19 cases of breast pain, and 12 cases of breast enlargement associated with esomeprazole. A search of the World Health Organization's Global Pharmacovigilance Database found 114 cases of gynecomastia, 38 cases of galactorrhea, 56 cases of breast pain, and 28 cases of breast enlargement associated with esomeprazole.
One woman developed elevated serum prolactin and estradiol levels, along with bilateral galactorrhea, one week after starting esomeprazole 40 mg once daily for the treatment of reflux esophagitis. The galactorrhea disappeared 3 days after esomeprazole was discontinued, and prolactin and estradiol levels returned to normal 7 days after discontinuation. One month later, the patient resumed esomeprazole and experienced bilateral galactorrhea again. She then switched to lansoprazole, and the galactorrhea did not recur. For mothers who have established lactation, prolactin levels may not affect their ability to breastfeed. Protein Binding Esomeprazole binds to plasma proteins at a rate of 97%. This binding remains constant within a concentration range of 2 to 20 µmol/L. Discussion: Proton pump inhibitors (PPIs) are generally well-tolerated with mild and transient side effects such as headache and diarrhea; high intravenous doses (up to 240 mg daily) are also well-tolerated in the treatment of Zollinger-Ellison syndrome. However, due to a lack of relevant studies, the potential problems of long-term high-dose use remain unclear. [1] In this mouse study, the high dose of esomeprazole (300 mg/kg/day, orally) used in the prophylactic regimen was poorly tolerated, resulting in adverse reactions such as abdominal distension, lethargy, and increased mortality. Kaplan-Meier survival curves showed that the survival rate of the high-dose group was lower than that of the control group. [2] Mice tolerated the lower therapeutic dose of esomeprazole (30 mg/kg/day, orally) well, and no significant adverse reactions were reported. [2] Autopsy organ weight analysis showed no significant difference in lung, heart, and kidney weight between the esomeprazole treatment group and the solvent control group. The liver weight was slightly increased in the esomeprazole treatment group, but the difference was not statistically significant. [2]

[1]. Use of proton pump inhibitors as adjunct treatment for triple-negative breast cancers. An introductory study. J Pharm Pharm Sci. 2014;17(3):439-46.

[2]. Therapeutic Efficacy of Esomeprazole in Cotton Smoke-Induced Lung Injury Model. Front Pharmacol. 2017 Jan 26;8:16.

[3]. Esomeprazole: a clinical review. Am J Health Syst Pharm. 2002 Jul 15;59(14):1333-9.

Esomeprazole magnesium is a magnesium salt formed by the reaction of magnesium hydroxide with 2 molar equivalents of esomeprazole. It is a gastric acid secretion inhibitor used to treat gastroesophageal reflux disease (GERD), dyspepsia, peptic ulcers, and Zollinger-Ellison syndrome (ZE syndrome). It is an EC 3.6.3.10 (H(+)/K(+) exchange ATPase) inhibitor and an anti-ulcer drug. It contains the esomeprazole (1-) domain. Esomeprazole, marketed as Nexium, is a proton pump inhibitor (PPI) used to treat gastroesophageal reflux disease (GERD), protect the gastric mucosa to prevent recurrence of gastric ulcers or gastric damage caused by long-term use of nonsteroidal anti-inflammatory drugs (NSAIDs), and treat pathological hypersecretion states including ZE syndrome. It is also often used in combination with other antibiotics (such as [DB01060], [DB01211], and [DB00916]) in quadruple therapy for Helicobacter pylori infection. Its efficacy is similar to other PPIs (e.g., [DB00338], [DB00213], [DB00448], [DB05351], and [DB01129]). Esomeprazole is the S-isomer of [DB00338], which is a racemic mixture of the S- and R-enantiomers. In vitro studies have shown that esomeprazole inhibits gastric acid secretion to a similar degree as [DB00338], with no significant difference between the two. Esomeprazole inhibits gastric acid secretion by covalently binding to the cysteine sulfhydryl group on the (H+, K+)-ATPase enzyme on the secretory surface of gastric parietal cells, thus preventing the final step in gastric acid formation. This action results in the inhibition of both basal and stimulant gastric acid secretion, unaffected by stimuli. Because the binding of esomeprazole to (H+, K+)-ATPase is irreversible, new enzyme expression is required to restore gastric acid secretion; therefore, the acid-suppressing effect of esomeprazole lasts for more than 24 hours. Proton pump inhibitors (PPIs) such as esomeprazole have also been shown to inhibit the activity of dimethylarginine dimethylaminohydrolase (DDAH), an enzyme crucial for cardiovascular health. DDAH inhibition leads to the accumulation of the nitric oxide synthase inhibitor asymmetric dimethylarginine (ADMA), which is believed to be the reason why PPIs are associated with an increased risk of cardiovascular events in patients with unstable coronary syndromes. Due to their good safety profile and the fact that many PPIs are available over-the-counter without a prescription, they are currently widely used in North America. However, long-term use of PPIs (such as esomeprazole) may produce adverse effects, including increased susceptibility to bacterial infections (including Clostridium difficile infection of the gastrointestinal tract), reduced absorption of micronutrients such as iron and vitamin B12, and increased risk of hypomagnesemia and hypocalcemia, all of which may lead to osteoporosis and fractures in later life. Rapid discontinuation of PPIs (such as esomeprazole) may lead to rebound effects and short-term excessive secretion. To prevent rebound effects, the dose of esomeprazole should be slowly reduced or gradually tapered until discontinued. Esomeprazole magnesium is the magnesium salt of esomeprazole, which is the S-isomer of omeprazole and has proton pump inhibitor activity. In the acidic environment of parietal cells, esomeprazole is protonated and converted into an active achiral sulfonamide; the active sulfonamide forms one or more covalent disulfide bonds with the proton pump hydrogen potassium ATPase (H+/K+ ATPase), thereby inhibiting its activity and the secretion of H+ ions from parietal cells into the gastric lumen, the final step in gastric acid production. H+/K+ ATPase is an integrated membrane protein of gastric parietal cells. Esomeprazole is the S-isomer of omeprazole and has proton pump inhibitor activity. In the acidic environment of parietal cells, esomeprazole is protonated and converted into an active achiral sulfinamide; this active sulfinamide forms one or more covalent disulfide bonds with the proton pump hydrogen potassium ATPase (H+/K+ ATPase), thereby inhibiting its activity and preventing parietal cells from secreting H+ ions into the gastric lumen, the final step in gastric acid production. H+/K+ ATPase is an integrated membrane protein of gastric parietal cells. Omeprazole is an S-isomer. See also: esomeprazole (containing the active moiety); esomeprazole magnesium; naproxen (component).

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Drug Indications
Esomeprazole is indicated for the treatment of acid reflux disorders, including healing and maintenance therapy for erosive esophagitis, symptomatic gastroesophageal reflux disease (GERD), peptic ulcers, Helicobacter pylori eradication, prevention of gastrointestinal bleeding induced by nonsteroidal anti-inflammatory drugs (NSAIDs), and long-term treatment of pathological hypersecretion states, including Zollinger-Ellison syndrome.

FDA Label

Nexium Controls is indicated for the short-term treatment of reflux symptoms (e.g., heartburn and acid reflux) in adults.Mechanism of Action
Esomeprazole exerts its inhibitory effect on gastric acid by covalently binding to the cysteine sulfhydryl groups on the secretory surface (H+, K+)-ATPase enzymes of the gastric parietal cells. This action results in the inhibition of both basal and stimulant gastric acid secretion, independent of stimuli.
Because the binding of esomeprazole to (H+, K+)-ATPase is irreversible, new enzymes need to be expressed to restore gastric acid secretion; therefore, the antisecretory effect of esomeprazole lasts for more than 24 hours. Esomeprazole is a proton pump inhibitor that inhibits gastric acid secretion by specifically inhibiting H+/K+-ATPase in gastric parietal cells. The S- and R-isomers of omeprazole are protonated in the acidic environment of parietal cells and converted into the active inhibitor—a non-chiral sulfonamide. Esomeprazole reduces gastric acidity by specifically acting on the proton pump, blocking the final step in gastric acid formation. This effect is dose-related and can be observed at daily doses ranging from 20 to 40 mg. Esomeprazole is a proton pump inhibitor (PPI) belonging to the substituted benzimidazole class of drugs. Its mechanism of action is to inhibit gastric acid secretion by irreversibly inhibiting H+/K+ ATPase in gastric parietal cells. Clinically, it is used to treat gastroesophageal reflux disease, peptic ulcers, and functional dyspepsia. [1] This study explores its potential use as an adjuvant therapy for cancer. The proposed mechanism is that esomeprazole inhibits the proton pump (specifically gastric H+/K+ ATPase expressed in breast cancer cells), leading to intracellular acidification, which in turn inhibits cancer cell growth and increases their sensitivity to chemotherapeutic drugs such as doxorubicin. [1] This study is the first to demonstrate that gastric H+/K+ ATPase is expressed in triple-negative breast cancer cells (MDA-MB-468) and mediates their sensitivity to esomeprazole. This finding is innovative because it was previously thought that the expression of this pump was limited to gastric parietal cells. [1] Esomeprazole is a proton pump inhibitor (PPI) used clinically to treat acid-related diseases such as gastroesophageal reflux disease (GERD). This study explores its potential use in treating lung injury and fibrosis. [2]
The proposed mechanism involves the inhibition of DDAH, leading to an increase in the level of its substrate ADMA, which is an endogenous inhibitor of iNOS. This results in reduced iNOS activity and NO production, thereby alleviating nitrosation stress, inflammation, and fibrosis. Proton pump inhibitors (PPIs) have also been found to upregulate heme oxygenase-1 (HO-1) and may activate the Keap1/Nrf2 pathway, thereby exerting cytoprotective and antioxidant effects. [2]
This study distinguishes the effects of the drug in an acidic stomach (where it is activated to inhibit H+/K+ ATPase) from its potential effects in extragastric tissues at neutral pH (where it may be active in its previous drug form). The beneficial effects observed in a lung injury model (where mice physiologically do not experience reflux) support that PPIs have direct pharmacological effects in addition to inhibiting gastric acid secretion. [2]
Retrospective clinical studies cited in the introduction and discussion have shown that PPI use is associated with good prognosis in chronic lung diseases such as idiopathic pulmonary fibrosis (IPF) and chronic obstructive pulmonary disease (COPD). [2]

C17H19N3O3S-.MG+2

369.721

712.198

1198768-91-0

Esomeprazole;119141-88-7;Esomeprazole magnesium trihydrate;217087-09-7;Esomeprazole sodium;161796-78-7;Esomeprazole magnesium;161973-10-0;Esomeprazole potassium salt;161796-84-5;Esomeprazole hemistrontium;914613-86-8

9568613

Typically exists as solid at room temperature

0

14

10

49

453

2

CC1=CN=C(C(=C1OC)C)C[S@](=O)C2=NC3=C([N-]2)C=CC(=C3)OC.CC1=CN=C(C(=C1OC)C)C[S@](=O)C2=NC3=C([N-]2)C=CC(=C3)OC.[Mg+2]

KWORUUGOSLYAGD-YPPDDXJESA-N

InChI=1S/2C17H18N3O3S.Mg/c2*1-10-8-18-15(11(2)16(10)23-4)9-24(21)17-19-13-6-5-12(22-3)7-14(13)20-17;/h2*5-8H,9H2,1-4H3;/q2*-1;+2/t2*24-;/m00./s1

magnesium;5-methoxy-2-[(S)-(4-methoxy-3,5-dimethylpyridin-2-yl)methylsulfinyl]benzimidazol-1-ide

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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