| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
TRPC3 140 nM (IC50)
Epithelial sodium channel (ENaC) (IC50 = 400 nM, irreversible inhibitor). TRPP3 (polycystin-L) (IC50 = 140 nM in Ca2+ uptake assay). PPARgamma (inducer). |
|---|---|
| ln Vitro |
TRPP3, a cation channel that belongs to the transient receptor potential (TRP) superfamily, is triggered by Ca2+ and permeable to Ca2+, Na+, and K+. In spinal cord neurons, TRPP3 is involved in the control of pH-sensitive action potentials. In radiotracer uptake experiment, phenamil methanesulfonate (1 μM) reduces 45Ca2+ absorption. With an IC50 value of 0.28 μM in oocytes expressing TRPP3 or oocytes injected with H2O, it inhibits TRPP3-mediated Ca2+ transport[1]. The epithelial sodium channel (ENaC) is inhibited by phenamil methanesulfonate, a more powerful ENaC blocker than amiloride, with an IC50 of 400 nM (vs 776 nM for amiloride)[2]. In both human and ovine bronchial epithelia cells, phenolmethyl methanesulfonate suppresses baseline short-circuit currents with IC50 values of 75 and 116 nM, respectively[3]. In C3H10T1/2 cells, phenamil methanesulfonate (0–20 μM; 14 days) controls adipogenesis and increases the expression of PPARγ, Fabp4, lipoprotein lipase, and adipogenic genes in a concentration-dependent manner[4]. Methanesulfonate (0–20 μM; 7 or 14 days) boosts concentration-dependently the activity of alkaline phosphatase (ALP) in MC3T3–E1 cells and modifies their osteoblastic differentiation[4].
Phenamil methanesulfonate is a potent inhibitor of ENaC (IC50 = 400 nM) and TRPP3 (IC50 = 140 nM in Ca2+ uptake assay). It induces differentiation and PPARgamma expression through ETS variant 4 in mouse preadipocytes. It promotes osteogenesis by activating bone morphogenetic protein (BMP) signaling. |
| ln Vivo |
PAH caused by chronic hypoxia is lessened by phenamil methanesulfonate (subcutaneous injection; 15 or 30 mg/kg; 21 days; infusion rate of 1 ml/h). Furthermore, Phenamil reduces the mRNA levels of SMA, SM22, Id3, and Trb3 in the lung sample in rats under hypoxia or normoxia. Phenamil, however, has minimal effects on the pulmonary vasculature in a physiological setting[5].
No in vivo data is available for Phenamil methanesulfonate. However, ENaC inhibitors are used as diuretics, and their in vivo activity is well characterized. Phenamil has been used in vivo to study its effects on pulmonary arterial hypertension and bone formation, though these studies are often with the parent compound, not the methanesulfonate salt. |
| Enzyme Assay |
Recombinant human ENaC channels (alpha, beta, gamma subunits) are expressed in Xenopus oocytes or HEK293 cells. The compound is applied to the bath solution, and the amiloride-sensitive sodium current is measured using two-electrode voltage-clamp (TEVC) or patch-clamp techniques.
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| Cell Assay |
RT-PCR[4]
Cell Types: C3H10T1/2 cells Tested Concentrations: 0 μM and 20 μM Incubation Duration: 14 days Experimental Results: Increased PPARγ, Fabp4, and lipoprotein lipase (LPL) mRNA expression. MDCK or mpkCCDc14 cells (kidney collecting duct cells) are cultured on permeable supports to form polarized monolayers. The compound is added to the apical medium, and the transepithelial sodium transport is measured using an Ussing chamber. Its effect on PPARgamma expression is studied in 3T3-L1 preadipocytes. |
| Animal Protocol |
Animal/Disease Models: Male SD (Sprague-Dawley) rats[5]
Doses: 15 or 30 mg /kg Route of Administration: subcutaneous (sc) injection; 15 or 30 mg/kg; 21 days; infusion rate of 1 ml/h Experimental Results: decreased hypoxia-induced pulmonary hypertension and vascular remodeling. Phenamil methanesulfonate is formulated in a suitable vehicle (e.g., 5% DMSO in saline) and administered via intraperitoneal injection to mice. Its effects on blood pressure and renal function can be assessed. For bone studies, it may be injected locally or delivered via an implant. |
| ADME/Pharmacokinetics |
The compound is moderately soluble in DMSO and 2-hydroxypropyl-beta-cyclodextrin. Its half-life and bioavailability are not specified. It is likely to be rapidly cleared due to its charged groups, but as a research tool, its local effects are often studied.
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| Toxicity/Toxicokinetics |
As an ENaC inhibitor, high systemic doses can lead to hyperkalemia, metabolic acidosis, and hypotension. It is generally well-tolerated in vitro. In vivo, its toxicity is related to its pharmacology, and specific toxicological data for this salt is limited.
|
| References | |
| Additional Infomation |
Phenamil methanesulfonate is a research tool for studying sodium transport, ENaC biology, and related pathways (PPARgamma, BMP). It is a second-generation analog of amiloride with increased potency and reduced reversibility, making it a valuable tool for mechanistic studies.
|
| Molecular Formula |
C13H16CLN7O4S
|
|---|---|
| Molecular Weight |
401.83
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| Exact Mass |
401.067
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| CAS # |
1161-94-0
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| PubChem CID |
135419184
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| Appearance |
Light yellow to yellow solid powder
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| Boiling Point |
621.7ºC at 760 mmHg
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| Flash Point |
329.8ºC
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| Vapour Pressure |
2.22E-15mmHg at 25°C
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| LogP |
3.381
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
3
|
| Heavy Atom Count |
26
|
| Complexity |
492
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CS(=O)(=O)O.C1=CC=C(C=C1)N=C(N)NC(=O)C2=C(N=C(C(=N2)Cl)N)N
|
| InChi Key |
MHPIZTURFVSLTJ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C12H12ClN7O.CH4O3S/c13-8-10(15)19-9(14)7(18-8)11(21)20-12(16)17-6-4-2-1-3-5-6;1-5(2,3)4/h1-5H,(H4,14,15,19)(H3,16,17,20,21);1H3,(H,2,3,4)
|
| Chemical Name |
3,5-diamino-6-chloro-N-(N'-phenylcarbamimidoyl)pyrazine-2-carboxamide;methanesulfonic acid
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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 (e.g. under nitrogen), 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)
|
| Solubility (In Vitro) |
DMSO: 25 mg/mL (62.22 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.22 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 | 2.4886 mL | 12.4431 mL | 24.8861 mL | |
| 5 mM | 0.4977 mL | 2.4886 mL | 4.9772 mL | |
| 10 mM | 0.2489 mL | 1.2443 mL | 2.4886 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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