| Size | Price | |
|---|---|---|
| Other Sizes |
| Targets |
Human Endogenous Metabolite
ITACONIC ACID targets multiple pathways: (1) It inhibits the enzyme isocitrate lyase (ICL), a key enzyme of the glyoxylate cycle essential for bacterial growth. (2) It modulates the pentose phosphate pathway (PPP) by inhibiting 6-phosphogluconate dehydrogenase (6PGD) and succinate dehydrogenase (SDH). (3) It activates the transcription factor Nrf2 (nuclear factor erythroid 2-related factor 2) by alkylating Keap1, leading to antioxidant gene expression. (4) It also inhibits TNF-alpha, IL-6, and IL-1beta production in macrophages. |
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| ln Vitro |
ITACONIC ACID exhibits anti-inflammatory and antibacterial activity: (1) In RAW 264.7 macrophages, itaconate (0.5-5 mM) significantly reduces LPS-induced TNF-alpha, IL-6, and IL-1beta production by 30-70%. (2) It promotes the pentose phosphate pathway (PPP), increasing NADPH oxidase activity and ROS production. (3) It binds to Nrf2 via alkylation of Keap1 cysteine residues, stabilizing Nrf2 and promoting Nrf2 nuclear translocation, leading to increased expression of Nrf2 target genes (HO-1, NQO1). (4) As an antibacterial, itaconate inhibits the growth of M. tuberculosis and other bacteria that rely on the glyoxylate cycle, with MIC values of 1-5 mM for M. tuberculosis. (5) Itaconate also inhibits succinate dehydrogenase (SDH) (IC50 ~ 10-30 uM), contributing to succinate accumulation and modulation of the TCA cycle.
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| ln Vivo |
In vivo, itaconic acid and its derivative 4-octyl itaconate (4-OI) exhibit anti-inflammatory activity: (1) In a murine model of LPS-induced septic shock (C57BL/6 mice, 5-10 mg/kg LPS IP), administration of 4-OI (50-200 mg/kg IP, 1 h before LPS) significantly reduces serum TNF-alpha and IL-6 levels by 40-60% and improves survival. (2) In a DSS-induced colitis model, itaconate treatment reduces disease activity index and colon inflammation. (3) In ischemia-reperfusion injury models, itaconate reduces tissue damage by upregulating Nrf2-dependent antioxidant responses. (4) Endogenous itaconic acid is produced by macrophages in response to bacterial infection and plays a crucial protective role.
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| Enzyme Assay |
(1) Itaconic acid is an endogenous metabolite, not an enzyme/substrate for in vitro binding assays. For SDH enzyme activity inhibition assay: (2) incubate purified porcine SDH (0.1 U) with itaconic acid (1-1000 uM) in reaction buffer (50 mM potassium phosphate, 0.2 mM EDTA, 0.2 mM KCN, 20 mM succinate, 0.05 mM DCIP, pH 7.4) at 25degC for 10 min. (3) Measure the reduction of DCIP at 600 nm. (4) Calculate IC50 using dose-response curve. (5) For isocitrate lyase assay: incubate purified M. tuberculosis isocitrate lyase (0.1 ug) with itaconic acid (0.1-10 mM) in 100 mM MOPS buffer (pH 7.0) containing 2 mM MgCl2, 10 mM dithiothreitol, and 4 mM isocitrate. Measure the formation of glyoxylate by a colorimetric assay at 324 nm.
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| Cell Assay |
(1) Seed RAW 264.7 macrophages (5,000-10,000 cells/well) in 96-well plates overnight. (2) Pre-treat cells with itaconic acid (0.1-10 mM) or 4-octyl itaconate (0.1-100 uM) for 1-2 h. (3) Stimulate with LPS (1 ug/mL) for 6-24 h. (4) Collect supernatant for cytokine measurement by ELISA (TNF-alpha, IL-6, IL-1beta). (5) For Nrf2 activation: treat cells with 4-OI (0.1-100 uM) for 6 h, then lyse, perform Western blot for NQO1, HO-1, and Nrf2 in nuclear extracts. (6) For succinate measurement: after treatment, extract metabolites and quantify succinate by LC-MS. (7) For viability: add MTT or CCK-8 reagent, measure OD.
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| Animal Protocol |
(1) Use 6-8 week old male C57BL/6 mice (20-25 g). (2) For LPS-induced septic shock model: administer 4-octyl itaconate (4-OI) at 50-200 mg/kg IP or itaconic acid at 100-500 mg/kg IP (formulated in 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline) 1 h before LPS injection (5-10 mg/kg IP). (3) Collect blood 6 h post-LPS by cardiac puncture, centrifuge for serum, measure TNF-alpha and IL-6 by ELISA. (4) Monitor survival for 48 h. (5) For DSS colitis model: add 2.5-3% DSS to drinking water for 7 days to induce colitis, treat with 4-OI (100 mg/kg IP) daily. (6) Measure colon length, histological score, and cytokine levels in colon homogenates. (7) For tumor models: use xenograft-bearing mice, assess tumor growth inhibition with itaconate derivatives.
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| ADME/Pharmacokinetics |
Standard formulation for in vitro: dissolve itaconic acid in water or PBS (pH 7.4) at 100-500 mM stock, or in DMSO (≥ 250 mg/mL). For 4-octyl itaconate (4-OI), dissolve in DMSO at 100-200 mM stock. For in vivo: formulate 4-OI (50-200 mg/kg) in 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline for IP injection. Endogenous itaconic acid concentrations in macrophages: upon LPS stimulation, intracellular itaconate reaches 1-10 mM. Plasma half-life of 4-OI in mice: t1/2 ~ 2-4 h (IP, 100 mg/kg). Oral bioavailability of itaconic acid is poor (~10-20%) due to its charged nature.
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| Toxicity/Toxicokinetics |
In vitro toxicity: itaconic acid (0.5-10 mM) in RAW 264.7 or HEK293 cells shows no significant cytotoxicity by MTT assay for up to 72 h; IC50 > 20 mM. In vivo toxicity: Single IP dose of 4-octyl itaconate (4-OI) in mice up to 500 mg/kg shows no mortality or severe toxicity; MTD > 500 mg/kg. Repeated dosing (100 mg/kg IP daily for 7 days) is well tolerated with no weight loss or organ damage observed in liver and kidney histology. The compound is for research use only, not for human therapeutic use.
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| References | |
| Additional Infomation |
Itaconic acid is a dicarboxylic acid, a derivative of methacrylic acid, in which one methyl hydrogen is replaced by a carboxylic acid group. It is a metabolite of both fungi and humans. It is a dicarboxylic acid, an olefinic compound, and a dicarboxylic acid fatty acid. Its function is related to succinic acid. It is the conjugate acid of itaconic acid (2-). Itaconic acid is a metabolite found or produced in Escherichia coli (K12 strain, MG1655 strain). Itaconic acid has also been reported in potatoes, corn smut, and several other organisms with relevant data.
Itaconic acid is an endogenous metabolite produced by activated macrophages from the TCA cycle intermediate cis-aconitate via the enzyme IRG1 (immune responsive gene 1, also known as ACOD1). It is a key molecule in immunometabolism that links immune activation to metabolic reprogramming. In addition to its anti-inflammatory and antibacterial activities, itaconic acid is also used as an industrial monomer in the production of polymers, resins, and synthetic fibers. The 4-octyl itaconate (4-OI) ester is a cell-permeable derivative commonly used in research to study the effects of itaconate in cells and animals. This compound is not FDA-approved and is strictly for research use only. |
| Molecular Formula |
C5H6O4
|
|---|---|
| Molecular Weight |
130.10
|
| Exact Mass |
130.026
|
| CAS # |
97-65-4
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| Related CAS # |
Itaconic acid-13C5;2095777-38-9
|
| PubChem CID |
811
|
| Appearance |
White to off-white solid powder
|
| Density |
1.4±0.1 g/cm3
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| Boiling Point |
381.4±25.0 °C at 760 mmHg
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| Melting Point |
162-164 °C WITH DECOMP, ALSO REPORTED AS 172 °C
; 175 °C
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| Flash Point |
198.7±19.7 °C
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| Vapour Pressure |
0.0±1.9 mmHg at 25°C
|
| Index of Refraction |
1.498
|
| LogP |
0.38
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
9
|
| Complexity |
158
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C=C(CC(=O)O)C(=O)O
|
| InChi Key |
LVHBHZANLOWSRM-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C5H6O4/c1-3(5(8)9)2-4(6)7/h1-2H2,(H,6,7)(H,8,9)
|
| Chemical Name |
2-methylidenebutanedioic acid
|
| 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)
|
| Solubility (In Vitro) |
H2O :~27.78 mg/mL (~213.53 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 25 mg/mL (192.16 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 7.6864 mL | 38.4320 mL | 76.8640 mL | |
| 5 mM | 1.5373 mL | 7.6864 mL | 15.3728 mL | |
| 10 mM | 0.7686 mL | 3.8432 mL | 7.6864 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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