| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Ki: 4.5 μM (Fumarate hydratase)[1]
Fumarate Hydratase (FH). Fumarate hydratase-IN-2 sodium salt is a competitive inhibitor of FH, binding to the active site of the enzyme. FH (also known as fumarase) is a tumor suppressor; its loss or inhibition leads to the accumulation of fumarate, an oncometabolite. |
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| ln Vitro |
In cell-free enzyme assays, Fumarate hydratase-IN-2 sodium salt acts as a competitive inhibitor of FH with a Ki of 4.5 microM. It is cell-permeable, meaning it can cross the cell membrane to access its intracellular target. It exhibits nutrient-dependent cytotoxicity, meaning its toxicity to cells depends on the availability of metabolic substrates, often showing higher potency when cells are forced to rely on oxidative phosphorylation.
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| ln Vivo |
In vivo activity data for Fumarate hydratase-IN-2 sodium salt are limited, as it is primarily used as a cellular tool. By inhibiting FH, it induces the accumulation of fumarate. This oncometabolite can inhibit prolyl hydroxylases (PHDs), stabilizing HIF-1alpha and promoting a pseudo-hypoxic state. This tool is used to simulate the metabolic effects of hereditary leiomyomatosis and renal cell cancer (HLRCC) in cell culture models, though specific animal studies with this compound are less common.
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| Enzyme Assay |
For non-cellular FH inhibition assays, purified recombinant human fumarate hydratase is incubated with its substrate, fumarate, in an assay buffer (e.g., Tris-HCl, pH 8.0). Fumarate hydratase-IN-2 sodium salt is added at varying concentrations (1-100 uM). The reaction is initiated, and the conversion of fumarate to malate is monitored spectrophotometrically by measuring the decrease in absorbance at 240 nm (fumarate has a UV absorption peak). Ki values are calculated via Lineweaver-Burk plots.
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| Cell Assay |
For cellular metabolic assays, cancer cells (e.g., renal carcinoma or melanoma cells) are cultured in media with varying nutrient levels (e.g., high vs. low glucose/glutamine) to assess nutrient-dependent cytotoxicity. Cells are treated with Fumarate hydratase-IN-2 sodium salt (5-50 uM) for 24-72 hours. Cell viability is measured by MTT or CellTiter-Glo. Fumarate accumulation is measured by LC-MS/MS, and HIF-1alpha stabilization is confirmed by Western blot as a pharmacodynamic marker.
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| Animal Protocol |
For in vivo pharmacodynamics, Fumarate hydratase-IN-2 sodium salt would likely be formulated in a vehicle such as PBS (phosphate-buffered saline) or a solution containing a co-solvent like DMSO (≤5%) and administered intraperitoneally or intravenously to mice. However, published in vivo efficacy protocols for this specific compound are limited. Blood and tissues would be collected to measure fumarate levels, malate levels, and the fumarate/malate ratio via mass spectrometry to confirm target engagement.
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| ADME/Pharmacokinetics |
Fumarate hydratase-IN-2 sodium salt has a molecular weight of 405.29 g/mol. It is soluble in water and DMSO, making it suitable for both aqueous and organic formulations. For in vitro use, a 10 mM stock solution can be prepared in DMSO. Detailed pharmacokinetic parameters (half-life, Cmax, AUC) for this specific inhibitor are not extensively published in the available literature, as it is primarily used as a cellular probe rather than for systemic efficacy. The powder should be stored at -20degC, protected from light.
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| Toxicity/Toxicokinetics |
Detailed toxicological data for Fumarate hydratase-IN-2 sodium salt are limited. It is a research chemical and is not for human use. Standard laboratory safety precautions should be followed, including the use of gloves, a lab coat, and eye protection. Avoid inhalation and ingestion. The compound may be irritating to the eyes and skin. Its mechanism of action involves inhibiting a TCA cycle enzyme, which could be detrimental to highly metabolically active cells, but selective toxicity is conferred by the nutrient-dependent cytotoxicity phenotype.
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| References | |
| Additional Infomation |
Fumarate hydratase-IN-2 sodium salt is also known as Fumarase inhibitor 3. It is a chemical probe used to study the role of fumarate as an oncometabolite, as seen in HLRCC, a cancer syndrome caused by mutations in the FH gene. It is not an FDA-approved drug. It is a valuable tool for validating FH as a target for cancer therapy, particularly for tumors with intact FH function that can be pharmacologically converted to an "HLRCC-like" metabolic state.
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| Molecular Formula |
C25H25N2NAO4
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|---|---|
| Molecular Weight |
440.466777563095
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| Exact Mass |
440.171
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| CAS # |
2070009-45-7
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| PubChem CID |
121596089
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
32
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| Complexity |
740
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| Defined Atom Stereocenter Count |
2
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| SMILES |
CNC(=O)C[C@@H]1C(=O)N(C2=CCCC[C@@]12C(=O)[O-])CC3=CC=C(C=C3)C4=CC=CC=C4.[Na+]
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| InChi Key |
KOACFUAVCMBNNV-RGFBGPCLSA-M
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| InChi Code |
InChI=1S/C25H26N2O4.Na/c1-26-22(28)15-20-23(29)27(21-9-5-6-14-25(20,21)24(30)31)16-17-10-12-19(13-11-17)18-7-3-2-4-8-18;/h2-4,7-13,20H,5-6,14-16H2,1H3,(H,26,28)(H,30,31);/q;+1/p-1/t20-,25-;/m1./s1
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| Chemical Name |
sodium;(3S,3aR)-3-[2-(methylamino)-2-oxoethyl]-2-oxo-1-[(4-phenylphenyl)methyl]-3,4,5,6-tetrahydroindole-3a-carboxylate
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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: 10.66 mg/mL (24.20 mM)
H2O: 6.25 mg/mL (14.19 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 | 2.2703 mL | 11.3515 mL | 22.7030 mL | |
| 5 mM | 0.4541 mL | 2.2703 mL | 4.5406 mL | |
| 10 mM | 0.2270 mL | 1.1352 mL | 2.2703 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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