| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
IC50: 10.5 μM (TRPP3 channel)[1] NHE[2] Macropinocytosis[3]
EIPA targets multiple proteins: it inhibits the Na+/H+ exchanger (NHE), a plasma membrane protein involved in sodium and proton transport across cell membranes. It also acts as an inhibitor of the TRPP3 channel (a transient receptor potential channel) with an IC50 of 10.5 μM. Additionally, EIPA blocks macropinocytosis, a form of endocytosis that is dependent on NHE activity. Through these mechanisms, EIPA modulates intracellular pH, sodium homeostasis, and cellular uptake processes. |
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| ln Vitro |
In X, EIPA (100 μM, 30 min) inhibits the Ca2+ uptake mediated by TRPP3. Ovum laevis [1]. The basal Na+ current is reversibly inhibited by EIPA hydrochloride (10-100 μM) (IC50: 19.5 μM)[1]. In IEC-18 cells, EIPA (300 μM, 6 h) increases autophagy via NHE3 (Na+/H+-exchanger 3)[2]. EIPA (20 μM, 2 h) inhibits the large absorption of CA-PZ via macropinocytosis in both MIA PaCa-2 and HT-29 cells[3]. Zinc/kainate toxicity is attenuated by EIPA (30 μM, 3h) by reducing Zn2+ entrance in cerebellar granule neurons[4]. EIPA (5-100 μM, 48h) inhibits MKN28 cell growth by upregulating p21 expression[5]. The increase in COX-2 protein level generated by LPS is inhibited by EIPA (3 μM, 6 h)[7].
In vitro, EIPA inhibits Na+/H+ exchanger activity with an IC50 of 0.033 μg/mL. It inhibits TRPP3-mediated Ca²⁺ uptake in Xenopus oocytes at 100 μM. EIPA blocks macropinocytosis in various cell types, including cancer cells such as gastric cancer, colon carcinoma, and pancreatic carcinoma cells. These activities make it a useful tool for studying NHE function, TRPP3 channel physiology, and macropinocytosis-dependent processes like antigen uptake and nutrient scavenging in cancer. |
| ln Vivo |
EIPA (intravenous injection, 1 mg/kg) dose-dependently reduces I/R (ischemia/reperfusion)-induced renal dysfunction in ddY strain mice [6]. EIPA (orally, 10 mg/kg) inhibits LPS-induced inflammation in a balloon-type LPS-induced inflammation model [7].
In vivo, EIPA has been studied for potential therapeutic applications in cancer, cardiovascular diseases, and cystic fibrosis. By inhibiting NHE, it may modulate intracellular pH and sodium balance, affecting cell proliferation, migration, and survival in pathological conditions. Its ability to block macropinocytosis suggests potential utility in preventing nutrient uptake by cancer cells. However, detailed in vivo efficacy data is limited, and EIPA is primarily used as a research tool rather than a therapeutic agent. |
| Enzyme Assay |
Methods: Effects of EIPA on proliferation, pH(c), [Cl(-)](c) and expression of proteins regulating cell cycle and MAPKs were studied in MKN28 expressing NHE exposed to EIPA for 48 h.
Results: EIPA suppressed proliferation of MKN28 cells by causing G(0)/G(1) arrest without any significant effects on pH(c), but associated with reduction of [Cl(-)](c). Although EIPA alone had no effects on pH(c), EIPA co-applied with DIDS (an inhibitor of Cl(-)/HCO(3)(-) exchangers; i.e., anion exchanger (AE) and Na+-driven Cl(-)/HCO(3)(-) exchanger (NDCBE)) reduced pH(c), suggesting that DIDS-sensitive Cl(-)/HCO(3)(-) transporters such as AE and/or NDCBE keep pH(c) normal by stimulating HCO(3)(-) uptake coupled with Cl(-) release under an NHE-inhibited condition. EIPA-induced lowered [Cl(-)](c) up-regulated expression of p21associated with phosphorylation of MAPKs, suppressing proliferation associated with G(0)/G(1) arrest.
Conclusions: EIPA suppressed proliferation of MKN28 cells through up-regulation of p21 expression via reduction of [Cl(-)](c) as a result from DIDS-sensitive Cl(-)/HCO(3)(-) exchanger-mediated compensation for keeping pH(c) normal under an NHE-inhibited condition. This is the first study revealing that an NHE inhibitor suppressed the proliferation of cancer cells by reducing [Cl(-)](c) but not pH(c)[5].
In vitro enzyme/receptor binding assays for EIPA typically measure inhibition of Na+/H+ exchange activity using fluorescent pH-sensitive dyes (e.g., BCECF) in cells loaded with the dye. Cells are treated with EIPA at various concentrations, and the rate of intracellular pH recovery following an acid load is measured to calculate IC50 values. For TRPP3 channel inhibition, calcium flux assays using fluorescent calcium indicators (e.g., Fluo-4) in cells expressing TRPP3 are performed. |
| Cell Assay |
Cell Proliferation Assay[5]
Cell Types: MKN28 cells Tested Concentrations: 5, 10, 25, 50, and 100 μM Incubation Duration: 48 h Experimental Results: Inhibited cell proliferation in a dose- and time-dependent manner. Western Blot Analysis[2] Cell Types: IEC-18 cells Tested Concentrations: 300 μM Incubation Duration: 6 h Experimental Results: Increased total LC3-II protein levels and P62 flux. Increased ATG5, 7, 12 and P62 expression. Cellular assays for EIPA involve treating cultured cells with the compound at concentrations ranging from nanomolar to micromolar. For NHE inhibition, cells are loaded with BCECF-AM, and intracellular pH is measured fluorometrically following ammonium chloride pulse-induced acidification. For macropinocytosis inhibition, cells are treated with EIPA and then exposed to fluorescent dextran or other fluid-phase markers; uptake is quantified by flow cytometry or fluorescence microscopy. For TRPP3 studies, calcium imaging is performed. |
| Animal Protocol |
Animal/Disease Models: Male ddY strain mouse [6]
Doses: 1 mg/kg Route of Administration: intravenous (iv) (iv)injection Experimental Results: Reduce histological renal damage and improve the increase in renal ET-1 content caused by I/R. Animal/Disease Models: air bag type LPS-induced inflammation model [7] Doses: 10 mg/kg Route of Administration: Oral Experimental Results:Inhibited LPS-induced leukocyte infiltration into air bags. Suppresses the amount of PGE2 in ostomy bag fluid. In vivo animal studies for EIPA typically involve administration to rodents via intraperitoneal or intravenous injection. Models of cancer, inflammation, or cardiovascular disease may be used. Endpoints include tumor growth, inflammatory markers, or physiological parameters. However, EIPA is primarily used in vitro as a research tool, and comprehensive in vivo efficacy data is limited. Dosing and administration protocols would need to be optimized based on pharmacokinetic properties. |
| ADME/Pharmacokinetics |
Pharmacokinetic properties of EIPA are typical of amiloride derivatives. The compound is moderately soluble in DMSO, DMF, and ethanol, with limited aqueous solubility. It is cell-permeable and used in both cell culture and in vivo settings. As a small molecule, it likely undergoes hepatic metabolism and renal excretion. Detailed PK parameters (half-life, bioavailability, clearance) would need to be determined in preclinical species for specific experimental applications. EIPA is soluble in DMSO at 14 mg/mL.
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| Toxicity/Toxicokinetics |
Preclinical toxicity of EIPA is not extensively documented in publicly available sources. As an amiloride derivative, it may share some safety profile with amiloride, which is clinically used as a potassium-sparing diuretic. Potential adverse effects include hyperkalemia, gastrointestinal disturbances, and renal effects. At research concentrations, EIPA is generally well-tolerated in cell culture. Standard toxicology studies would be required for therapeutic development. Safety pharmacology would assess cardiovascular and renal effects.
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| References | |
| Additional Infomation |
Ethylisopropylamiloride amiloride belongs to the pyrazine class of compounds. Its structure is similar to amiloride, except that the amino group adjacent to the chlorine substituent on the pyrazine ring is replaced by an ethyl or isopropyl group. It possesses antiarrhythmic, neuroprotective, and sodium channel blocking effects. It belongs to the guanidine, aromatic amine, organochlorine, tertiary amine, pyrazine, and monocarboxylic acid amide classes. Its function is related to that of amiloride.
EIPA (Ethylisopropylamiloride) is a research-grade inhibitor of Na+/H+ exchanger (NHE) and macropinocytosis. It also inhibits the TRPP3 channel. It is used in studies of ion transport, fluid balance, inflammation, and cancer, including gastric cancer, colon carcinoma, and pancreatic carcinoma. EIPA is available for research purposes only and is not approved for clinical use. |
| Molecular Formula |
C11H18CLN7O
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|---|---|
| Molecular Weight |
299.76
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| Exact Mass |
299.126
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| Elemental Analysis |
C, 44.08; H, 6.05; Cl, 11.83; N, 32.71; O, 5.34
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| CAS # |
1154-25-2
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| Related CAS # |
EIPA hydrochloride;1345839-28-2
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| PubChem CID |
1795
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| Appearance |
Typically exists as White to yellow solids at room temperature
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| Density |
1.5±0.1 g/cm3
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| Boiling Point |
543.9±60.0 °C at 760 mmHg
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| Melting Point |
202-205ºC(lit.)
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| Flash Point |
282.8±32.9 °C
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| Vapour Pressure |
0.0±1.5 mmHg at 25°C
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| Index of Refraction |
1.665
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| LogP |
4.86
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
20
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| Complexity |
372
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C(C1=NC(Cl)=C(N(CC)C(C)C)N=C1N)NC(N)=N
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| InChi Key |
QDERNBXNXJCIQK-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C11H18ClN7O/c1-4-19(5(2)3)9-7(12)16-6(8(13)17-9)10(20)18-11(14)15/h5H,4H2,1-3H3,(H2,13,17)(H4,14,15,18,20)
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| Chemical Name |
3-amino-6-chloro-N-(diaminomethylidene)-5-[ethyl(propan-2-yl)amino]pyrazine-2-carboxamide
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| Synonyms |
Ethyl isopropylamiloride; EIPA; 1154-25-2; 5-(N-Ethyl-N-isopropyl)amiloride; Ethylisopropylamiloride; 5-(Ethylisopropyl)amiloride; Ethyl isopropyl amiloride; 5-(N-Ethyl-N-isopropyl) Amiloride; 5-(N-ethyl-N-isopropyl)-Amiloride;
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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 |
| 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 (~166.80 mM)
H2O : < 0.1 mg/mL |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.34 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.3360 mL | 16.6800 mL | 33.3600 mL | |
| 5 mM | 0.6672 mL | 3.3360 mL | 6.6720 mL | |
| 10 mM | 0.3336 mL | 1.6680 mL | 3.3360 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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