| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| Other Sizes |
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| Targets |
IC50: 10.5 μM (TRPP3 channel)[1] NHE[2] Macropinocytosis[3]
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|---|---|
| ln Vitro |
In X, TRPP3-mediated Ca2+ uptake is inhibited by EIPA hydrochloride (100 μM, 30 min). Ovum laevis [1]. Basal Na+ current is reversibly inhibited by EIPA hydrochloride (10-100 μM) (IC50: 19.5 μM) [1]. Through NHE3 (Na+/H+-exchanger 3), EIPA hydrochloride (300 μM, 6 hours) increases autophagy in IEC-18 cells [2]. The uptake of CA-PZ into HT-29 cells and MIA PaCa-2 cells through macropinocytosis is inhibited by EIPA hydrochloride (20 μM, 2 hours) [3]. Zinc/kainic acid toxicity is lessened by EIPA hydrochloride (30 μM, 3 hours) by decreasing Zn2+ entry into cerebellar granule neurons [4]. By upregulating p21 expression, EIPA hydrochloride (5-100 μM, 48 hours) inhibits MKN28 cell proliferation [5]. EIPA hydrochloride (3 μM, 6 hours) prevents the rise in COX-2 protein levels brought on by LPS [7].
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| ln Vivo |
In ddY strain mice, renal impairment caused by I/R (ischemia/reperfusion) is dose-dependently reduced by intravenous administration of 1 mg/kg of EIPA hydrochloride [6]. ?In the balloon-type LPS-induced inflammation paradigm, EIPA hydrochloride (10 mg/kg) reduces LPS-induced inflammation [7].
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| 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].
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| Cell Assay |
Cell proliferation assay [5]
Cell Types: MKN28 Cell Tested Concentrations: 5, 10, 25, 50 and 100 μM Incubation Duration: 48 h Experimental Results: Inhibition of cell proliferation in a dose- and time-dependent manner. Western Blot Analysis[2] Cell Types: IEC-18 Cell Tested Concentrations: 300 μM Incubation Duration: 6 hrs (hours) Experimental Results: Increased LC3-II total protein levels and P62 flux. The expression of ATG5, 7, 12 and P62 was increased. |
| 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. |
| References |
[1]. Inhibition of TRPP3 channel by MK-870 and analogs. Mol Pharmacol. 2007 Dec;72(6):1576-85.
[2]. Na+/H+ Exchanger Regulates Amino Acid-Mediated Autophagy in Intestinal Epithelial Cells. Cell Physiol Biochem. 2017;42(6):2418-2429. [3]. A new HDAC inhibitor cinnamoylphenazine shows antitumor activity in association with intensive macropinocytosis.Oncotarget. 2017 Feb 28;8(9):14748-14758. [4]. Acidosis and 5-(N-ethyl-N-isopropyl)amiloride (EIPA) Attenuate Zinc/Kainate Toxicity in Cultured Cerebellar Granule Neurons. Biochemistry (Mosc). 2015 Aug;80(8):1065-72. [5]. An inhibitor of Na(+)/H(+) exchanger (NHE), ethyl-isopropyl amiloride (EIPA), diminishes proliferation of MKN28 human gastric cancer cells by decreasing the cytosolic Cl(-) concentration via DIDS-sensitive pathways. Cell Physiol Biochem. 2012;30(5):1241-53. [6]. Role of Na+/H+ exchanger in the pathogenesis of ischemic acute renal failure in mice. J Cardiovasc Pharmacol. 2007 Mar;49(3):154-60. [7]. Inhibition of lipopolysaccharide-induced prostaglandin E2 production and inflammation by the Na+/H+ exchanger inhibitors. J Pharmacol Exp Ther. 2007 Apr;321(1):345-52. |
| Additional Infomation |
TRPP3, a member of the transient receptor potential (TRP) superfamily of cation channels, is a Ca2+-activated channel permeable to Ca2+, Na+, and K+. TRPP3 has been implicated in sour tasting in bipolar cells of tongue and in regulation of pH-sensitive action potential in spinal cord neurons. TRPP3 is also present in excitable and nonexcitable cells of other tissues, including retina, brain, heart, testis, and kidney, with unknown functions. In this study, we examined the functional modulation of TRPP3 channel by amiloride and its analogs, known to inhibit several ion channels and transporters and respond to all taste stimuli, using Xenopus laevis oocyte expression, electrophysiology, and radiotracer measurements. We found that amiloride and its analogs inhibit TRPP3 channel activities with different affinities. Radiolabeled (45)Ca2+ uptake showed that TRPP3-mediated Ca2+ transport was inhibited by amiloride, phenamil, benzamil, and 5-(N-ethyl-N-isopropyl)amiloride (EIPA). Two-microelectrode voltage clamp experiments revealed that TRPP3-mediated Ca2+-activated currents are substantially inhibited by amiloride analogs, in an order of potency of phenamil > benzamil > EIPA > amiloride, with IC50 values of 0.14, 1.1, 10.5, and 143 microM, respectively. The inhibition potency positively correlated with the size of inhibitors. Using cell-attached patch clamping, we showed that the amiloride analogs decrease the open probability and mean open time but have no effect on single-channel conductance. Study of inhibition by phenamil in the presence of previously reported inhibitor tetrapentylammonium indicates that amiloride and organic cation inhibitors compete for binding the same site on TRPP3. TRPP3 may contribute to previously reported in vivo amiloride-sensitive cation transport.[1]
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| Molecular Formula |
C11H19CL2N7O
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|---|---|
| Molecular Weight |
336.220858812332
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| Exact Mass |
335.1
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| Elemental Analysis |
C, 39.30; H, 5.70; Cl, 21.09; N, 29.16; O, 4.76
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| CAS # |
1345839-28-2
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| Related CAS # |
EIPA;1154-25-2
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| PubChem CID |
56935648
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| Appearance |
Typically exists as Light yellow to yellow solids at room temperature
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
21
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| Complexity |
372
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
BGHBOQNLQNKPFO-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C11H18ClN7O.ClH/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);1H
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| Chemical Name |
3-amino-6-chloro-N-(diaminomethylidene)-5-[ethyl(propan-2-yl)amino]pyrazine-2-carboxamide;hydrochloride
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| Synonyms |
EIPA hydrochloride; EIPA (hydrochloride); 1345839-28-2; CHEMBL1909809; 3-amino-6-chloro-N-(diaminomethylidene)-5-[ethyl(propan-2-yl)amino]pyrazine-2-carboxamide;hydrochloride; L593754 hydrochloride; 5-(N-Ethyl-N-isopropyl)amiloridehydrochloride; MH 12-43 hydrochloride; .
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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: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 : ~130 mg/mL (~386.65 mM)
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|---|---|
| Solubility (In Vivo) |
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.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] Oral Formulations
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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.9742 mL | 14.8712 mL | 29.7424 mL | |
| 5 mM | 0.5948 mL | 2.9742 mL | 5.9485 mL | |
| 10 mM | 0.2974 mL | 1.4871 mL | 2.9742 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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