| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| Other Sizes |
| Targets |
EWP-815 targets several enzymes. It is a potent inhibitor of Ins(1,4)P2 phosphatase and Ins(1,4,5)P3 5-phosphatase. These enzymes are involved in the metabolism of inositol phosphates, which are important second messengers in cellular signaling pathways. By inhibiting these phosphatases, EWP-815 affects the levels of inositol phosphates and downstream signaling. The compound also inhibits dopamine beta-hydroxylase, an enzyme involved in the synthesis of the neurotransmitter norepinephrine. This makes it a valuable tool for studying both inositol phosphate signaling and catecholamine metabolism.
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| ln Vitro |
In prelabeled GH3 cells, EWP 815 decreases the basal breakdown rate without influencing the thyrotropin-releasing hormone (TRH; 0.03 mM)-stimulated inositol phospholipid breakdown [1]. The soluble enzyme IC50 is 83 mM, whereas the granzyme is 71 mM. With an average [3H]Ins(1,4)P2 recovery of 2.6[1], EWP 815 (30 μM; 1 hour) suppresses Ins(1,4)P2 phosphatase more than Ins(1,4,5)P3 5-phosphatase. With an IC50 of 6 μM and 8 μM, respectively, EWP 815 (6, 8 μM; 10 min) inhibits the dephosphorylation of [3H]Ins(1,4,5)P3 5-phosphatase in both the soluble and particulate fractions[1]. Without changing Ins (1,4,5)P3 binding, EWP 815 (3-300 μM; 30 min) reduces the amount of inositol phospholipid produced in response to thyrotropin-releasing hormone (TRH) stimulation (100 nM) by 30% at 100 μM and 1.8% at 300 μM, respectively[1].
In vitro, EWP-815 has been shown to reduce the amount of inositol phospholipid produced in response to thyrotropin-releasing hormone (TRH) stimulation. In a study, EWP-815 (3-300 µM; 30 min) reduced the amount of inositol phospholipid produced in response to TRH stimulation (100 nM) by 30% at 100 µM and 1.8% at 300 µM. This effect was observed without changing Ins(1,4,5)P3 binding, indicating that the compound acts by inhibiting the phosphatases involved in its metabolism rather than by blocking the receptor. These findings highlight its role as a phosphatase inhibitor. |
| ln Vivo |
Mice treated intraperitoneally with EWP 815 (50 mg/kg; 30 minutes prior to sacrifice) have their dopamine β-hydroxylase activity inhibited [2].
Specific in vivo activity data for EWP-815 are not detailed in the available literature. As a research chemical, its primary application is for in vitro studies to investigate enzyme inhibition. Its potential in vivo effects would be related to its inhibition of inositol phosphate metabolism and dopamine beta-hydroxylase. Further studies in animal models would be needed to confirm its in vivo efficacy, pharmacokinetics, and safety. The compound is intended for research use only. |
| Enzyme Assay |
In vitro enzyme assays for EWP-815 involve measuring the inhibition of Ins(1,4)P2 phosphatase, Ins(1,4,5)P3 5-phosphatase, or dopamine beta-hydroxylase. For phosphatase assays, the enzyme is incubated with its substrate (e.g., Ins(1,4,5)P3) in a buffer solution. Varying concentrations of EWP-815 are added to the reaction mixture, and the hydrolysis of the substrate is measured. For dopamine beta-hydroxylase assays, the enzyme is incubated with its substrate (dopamine) and cofactors, and the formation of norepinephrine is measured. The compound's inhibitory activity is determined by calculating the IC50 or by measuring the percentage of inhibition at a given concentration.
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| Cell Assay |
In vitro cell-based assays for EWP-815 are performed to study its effects on inositol phosphate signaling and neurotransmitter metabolism. Cells are labeled with [3H]-inositol to track the production of inositol phosphates. The cells are then stimulated with an agonist (e.g., TRH) in the presence or absence of varying concentrations of EWP-815. The inositol phosphates are extracted and separated by chromatography, and the amount of labeled inositol phosphates is measured. The compound's effect on dopamine beta-hydroxylase activity can be studied in cells that express the enzyme by measuring the conversion of dopamine to norepinephrine.
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| Animal Protocol |
Animal/Disease Models: Female mice (20 g) [2]
Doses: 50 mg/kg Route of Administration: intraperitoneal (ip) injection; 30 minutes before sacrifice Hydroxylase. The amount of 3H-α-Me-NA was diminished by 12%. Specific in vivo animal experiment protocols for EWP-815 are not detailed in the available literature. For potential in vivo studies, the compound can be formulated for administration. The route of administration (e.g., oral, intraperitoneal, or intravenous) and dosing regimen would depend on the specific experimental design and target being studied. Given its mechanism of action, it could be used in models of neurological or signaling disorders. |
| ADME/Pharmacokinetics |
EWP-815 has a molecular weight of 350.6 g/mol and the formula C12H22N4S4. It is chemically known as bis(4-methyl-1-piperazinylthiocarbonyl) disulfide. The compound is a disulfide analog and is intended for research use only. Its pharmacokinetic properties, such as absorption, distribution, metabolism, and excretion, have not been extensively characterized in the available literature. For long-term storage, the compound is kept as a powder under appropriate conditions.
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| Toxicity/Toxicokinetics |
Specific toxicological data for EWP-815 are not provided in the available sources. As a research chemical, its safety profile in humans has not been established. The compound is classified as a research reagent and is not for therapeutic or veterinary use. Standard laboratory safety precautions should be followed when handling this compound, including the use of appropriate personal protective equipment.
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| References | |
| Additional Infomation |
EWP-815 is a synthetic disulfiram analogue and a potent inhibitor of key enzymes involved in cellular signaling and neurotransmitter metabolism. Chemically known as bis(4-methyl-1-piperazinylthiocarbonyl) disulfide, it has the molecular formula C12H22N4S4 and a molecular weight of 350.6 g/mol. EWP-815 is a potent inhibitor of Ins(1,4)P2 phosphatase and Ins(1,4,5)P3 5-phosphatase. It also inhibits dopamine beta-hydroxylase activity. In vitro, it has been shown to reduce inositol phospholipid production in response to TRH stimulation. The compound is used in research to study inositol phosphate signaling and neurotransmitter metabolism and is intended for research use only.
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| Molecular Formula |
C12H22N4S4
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| Molecular Weight |
350.58988
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| Exact Mass |
350.073
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| CAS # |
20231-01-0
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| PubChem CID |
30054
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| Appearance |
White to off-white solid powder
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| Density |
1.339g/cm3
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| Boiling Point |
450.5ºC at 760 mmHg
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| Melting Point |
142ºC
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| Flash Point |
226.3ºC
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| Vapour Pressure |
2.62E-08mmHg at 25°C
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| Index of Refraction |
1.673
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| LogP |
1.184
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
20
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| Complexity |
315
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
OOBDBFPYJXJEHC-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C12H22N4S4/c1-13-3-7-15(8-4-13)11(17)19-20-12(18)16-9-5-14(2)6-10-16/h3-10H2,1-2H3
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| Chemical Name |
(4-methylpiperazine-1-carbothioyl)sulfanyl 4-methylpiperazine-1-carbodithioate
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~50 mg/mL (~142.62 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.25 mg/mL (3.57 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 12.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 2: ≥ 1.25 mg/mL (3.57 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 12.5 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 | 2.8523 mL | 14.2617 mL | 28.5233 mL | |
| 5 mM | 0.5705 mL | 2.8523 mL | 5.7047 mL | |
| 10 mM | 0.2852 mL | 1.4262 mL | 2.8523 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.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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