| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
4-Anilino-4-oxobutanoic acid-d5 is a deuterium-labeled analytical standard and does not directly target biological receptors. The non-labeled parent compound, 4-Anilino-4-oxobutanoic acid (succinanilic acid), is a metabolite of the histone deacetylase (HDAC) inhibitor suberoylanilide hydroxamic acid (SAHA, vorinostat). Vorinostat exerts its anticancer effects by inhibiting HDACs, leading to the accumulation of acetylated histones and other proteins, which alters gene expression and induces cell cycle arrest, differentiation, and apoptosis. The metabolism of vorinostat involves glucuronidation and hydrolysis to aniline-containing metabolites, including 4-anilino-4-oxobutanoic acid. This metabolite is considered inactive and is excreted in the urine. The deuterated version is used solely as an analytical standard for quantifying the metabolite in pharmacokinetic studies and does not engage in biological interactions.
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| ln Vitro |
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as quantitative tracers while the drugs were being developed. Because deuteration may have an effect on a drug's pharmacokinetics and metabolic properties, it is a cause for concern [1].
4-Anilino-4-oxobutanoic acid-d5, as a deuterium-labeled internal standard, does not possess intrinsic in vitro biological activity. It is used exclusively for analytical quantification purposes. The non-labeled 4-Anilino-4-oxobutanoic acid (succinanilic acid) is a metabolite of vorinostat (SAHA) and is not known to have significant biological activity. In vitro, vorinostat (the parent drug) inhibits HDAC activity, with IC50 values in the low nanomolar range (e.g., 10-100 nM) for HDAC1, HDAC2, HDAC3, and HDAC6. This leads to the accumulation of acetylated histones and the induction of cell cycle arrest and apoptosis in cancer cells. The metabolite 4-Anilino-4-oxobutanoic acid does not inhibit HDACs at physiologically relevant concentrations. The deuterated version is used as an internal standard to accurately quantify the metabolite in cell culture supernatants or lysates. |
| ln Vivo |
4-Anilino-4-oxobutanoic acid-d5 does not exhibit in vivo biological activity because it is an analytical standard. The non-labeled 4-Anilino-4-oxobutanoic acid is a major metabolite of vorinostat (SAHA). After oral administration of vorinostat, it is extensively metabolized, and 4-anilino-4-oxobutanoic acid is a prominent metabolite found in plasma and urine. The metabolite is considered inactive and does not contribute to the therapeutic effects of vorinostat. In vivo, vorinostat (the parent drug) has demonstrated antitumor activity in patients with cutaneous T-cell lymphoma and is being investigated for other cancers. The plasma concentration of 4-anilino-4-oxobutanoic acid is often measured in pharmacokinetic studies to assess the metabolism of vorinostat. As an internal standard, the deuterated form is used to accurately quantify the metabolite in plasma, urine, and tissue samples.
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| Enzyme Assay |
A standard non-cellular protocol for using 4-Anilino-4-oxobutanoic acid-d5 as an internal standard involves its inclusion in the sample preparation workflow for LC-MS analysis. A stock solution of the internal standard (1 mg/mL) is prepared in methanol or acetonitrile. For plasma samples, 50 uL of plasma is transferred to a microcentrifuge tube. Then, 10 uL of the internal standard solution (diluted to a working concentration of 1 ug/mL) is added. Proteins are precipitated by adding 150 uL of ice-cold acetonitrile containing 0.1% formic acid. The mixture is vortexed for 1 minute and centrifuged at 14,000 rpm for 10 minutes at 4degC. The supernatant (150 uL) is transferred to an autosampler vial and mixed with 150 uL of water containing 0.1% formic acid. The sample (5-10 uL) is injected onto an LC-MS/MS system operated in negative or positive ion mode (depending on the compound's ionization properties) with multiple reaction monitoring (MRM) for the specific transitions of 4-anilino-4-oxobutanoic acid and the deuterated internal standard. Quantification is achieved by calculating the peak area ratio of the analyte to the internal standard against a calibration curve.
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| Cell Assay |
A typical in vitro cellular protocol for using 4-Anilino-4-oxobutanoic acid-d5 as an internal standard involves the quantification of the metabolite in cultured cancer cells. Human cancer cell lines (e.g., HCT-116 colon cancer cells) are seeded in 6-well plates at a density of 5×10⁵ cells/well and cultured overnight. Cells are treated with vorinostat (non-labeled, 0.1-10 uM) for 24-48 hours. After treatment, the culture medium is collected. Cells are washed twice with ice-cold PBS and harvested by scraping. Cell pellets are resuspended in 200 uL of PBS, and 10 uL of the internal standard solution (1 ug/mL) is added. Cells are lysed by sonication (3×10 seconds on ice). Proteins are precipitated by adding 800 uL of acetonitrile containing 0.1% formic acid. After centrifugation, the supernatant is analyzed by LC-MS/MS. The concentration of 4-anilino-4-oxobutanoic acid in the culture medium and cell lysate is quantified using a calibration curve prepared with the same internal standard. The internal standard corrects for extraction efficiency and matrix effects.
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| Animal Protocol |
A typical in vivo animal protocol for using 4-Anilino-4-oxobutanoic acid-d5 as an internal standard involves the quantification of the metabolite in plasma and tissues. Male BALB/c nude mice bearing subcutaneous tumor xenografts (e.g., HCT-116) are administered vorinostat (non-labeled) via intraperitoneal injection at a dose of 50-100 mg/kg. Blood samples (50-100 uL) are collected via the tail vein into EDTA-coated tubes at various time points (0, 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24 hours) post-dose. Plasma is separated by centrifugation (2,000 g, 10 minutes, 4degC). For tissue analysis, mice are euthanized at the final time point, and tumors, liver, kidney, and other tissues are harvested, weighed, and homogenized in PBS. For extraction, 50 uL of plasma or tissue homogenate is mixed with 10 uL of 4-Anilino-4-oxobutanoic acid-d5 internal standard solution (1 ug/mL) and 150 uL of acetonitrile. After centrifugation, the supernatant is analyzed by LC-MS/MS. The internal standard corrects for variations in sample preparation and matrix effects.
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| ADME/Pharmacokinetics |
As an analytical internal standard, 4-Anilino-4-oxobutanoic acid-d5 is not characterized by typical pharmacokinetic parameters. However, the non-labeled 4-Anilino-4-oxobutanoic acid is a major metabolite of vorinostat (SAHA). After oral or intravenous administration of vorinostat, it is rapidly converted to this metabolite. The plasma concentration of the metabolite often exceeds that of the parent drug. The metabolite is formed via hydrolysis of the hydroxamic acid group and is further metabolized by glucuronidation. It is eliminated primarily in the urine. The half-life of the metabolite in humans is approximately 2-3 hours. The deuterated internal standard is used to accurately quantify the metabolite in biological samples for pharmacokinetic and metabolism studies of vorinostat.
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| Toxicity/Toxicokinetics |
Toxicity data specific to 4-Anilino-4-oxobutanoic acid-d5 are not available. The non-labeled 4-Anilino-4-oxobutanoic acid is a metabolite of vorinostat (SAHA). Vorinostat has a known toxicity profile, including gastrointestinal disturbances (nausea, vomiting, diarrhea), fatigue, thrombocytopenia, and anemia. The metabolite itself is considered less active and is not known to be toxic at concentrations typically observed in vivo. Standard laboratory safety precautions should be followed when handling the compound, including the use of gloves, lab coats, and safety glasses. Avoid inhalation, ingestion, and direct skin or eye contact. The compound should be stored at -20degC or as recommended by the supplier, protected from light and moisture. It is intended for research use only and should not be used in humans or animals for therapeutic or diagnostic purposes.
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| References | |
| Additional Infomation |
4-Anilino-4-oxobutanoic acid-d5 (CAS# 840529-98-8) is a stable isotope-labeled compound with a molecular weight of 198.23. The molecular formula is C10H6D5NO3, and it is also known as succinanilic acid-d5 and d5-4-anilino-4-oxobutanoic acid. The compound is a labelled analogue of 4-anilino-4-oxobutanoic acid, which is the metabolite (MII) of suberoylanilide hydroxamic acid (SAHA, vorinostat). Vorinostat is a histone deacetylase (HDAC) inhibitor used in the treatment of cutaneous T-cell lymphoma. The deuterated version is intended for use as an internal standard for the quantification of 4-anilino-4-oxobutanoic acid by LC-MS. This product is for research use only and is not approved for clinical diagnostic applications. The compound should be stored at -20degC, protected from light and moisture.
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| Molecular Formula |
C10H6D5NO3
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| Molecular Weight |
198.23
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| Exact Mass |
198.105
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| CAS # |
840529-98-8
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| PubChem CID |
45038175
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| Appearance |
White to off-white solid powder
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| Density |
1.32g/cm3
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| Boiling Point |
462.29ºC at 760 mmHg
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| Melting Point |
146-149°C
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| Flash Point |
233.385ºC
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| Index of Refraction |
1.6
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| LogP |
2.139
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
14
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| Complexity |
209
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| Defined Atom Stereocenter Count |
0
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| SMILES |
[2H]C1=C(C(=C(C(=C1[2H])[2H])NC(=O)CCC(=O)O)[2H])[2H]
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| InChi Key |
KTFGFGGLCMGYTP-RALIUCGRSA-N
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| InChi Code |
InChI=1S/C10H11NO3/c12-9(6-7-10(13)14)11-8-4-2-1-3-5-8/h1-5H,6-7H2,(H,11,12)(H,13,14)/i1D,2D,3D,4D,5D
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| Chemical Name |
4-oxo-4-(2,3,4,5,6-pentadeuterioanilino)butanoic 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)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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 | 5.0446 mL | 25.2232 mL | 50.4465 mL | |
| 5 mM | 1.0089 mL | 5.0446 mL | 10.0893 mL | |
| 10 mM | 0.5045 mL | 2.5223 mL | 5.0446 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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