| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
N-Acetyl mesalazine-d3, as an isotopically labeled internal standard, does not target specific biological receptors. The non-labeled parent compound, N-Acetyl mesalazine (N-Acetyl-5-aminosalicylic acid), is a metabolite of the anti-inflammatory drug mesalazine (5-ASA). Its mechanism of action is primarily related to its anti-inflammatory and antioxidant properties. N-Acetyl mesalazine can scavenge free radicals, reduce DNA base hydroxylation, and ameliorate mucosal inflammation. It is also an inhibitor of recombinant human thiopurine methyltransferase (hTPMT), an enzyme that methylates thiopurine drugs such as azathioprine and 6-mercaptopurine. By inhibiting hTPMT, N-Acetyl mesalazine may alter the metabolism and toxicity of thiopurine drugs. The deuterated version is used solely as an analytical standard 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].
As a deuterium-labeled internal standard, N-Acetyl mesalazine-d3 itself does not possess intrinsic in vitro biological activity. However, the non-labeled N-Acetyl mesalazine has demonstrated in vitro anti-inflammatory and antioxidant activities. In cell-free assays, N-Acetyl mesalazine scavenges free radicals such as DPPH and superoxide anions, with an IC50 in the micromolar range. It also reduces DNA base hydroxylation induced by reactive oxygen species. In cultured intestinal epithelial cells, N-Acetyl mesalazine reduces the production of pro-inflammatory cytokines (e.g., IL-6, IL-8, TNF-alpha) and chemokines in response to inflammatory stimuli. It also inhibits the activity of recombinant human thiopurine methyltransferase (hTPMT) in enzyme activity assays, with an IC50 of approximately 50 uM. The compound's anti-inflammatory effects are mediated in part by the inhibition of NF-kappaB activation and the downregulation of cyclooxygenase-2 (COX-2) expression. |
| ln Vivo |
N-Acetyl mesalazine-d3 does not exhibit in vivo biological activity because it is an analytical standard. The non-labeled N-Acetyl mesalazine is the major metabolite of mesalazine (5-ASA), an anti-inflammatory drug used to treat inflammatory bowel disease (IBD). After oral administration of mesalazine, it is acetylated to N-Acetyl mesalazine in the intestinal mucosa and liver. N-Acetyl mesalazine is considered to have lower anti-inflammatory activity compared to mesalazine, but it may still contribute to the therapeutic effects of mesalazine. In addition, N-Acetyl mesalazine has been shown to inhibit thiopurine methyltransferase (TPMT) activity in vivo, which can alter the pharmacokinetics and toxicity of thiopurine drugs such as azathioprine and 6-mercaptopurine, which are often used in combination with mesalazine to treat IBD. As an internal standard, the deuterated form is used to accurately quantify N-Acetyl mesalazine in plasma and urine for pharmacokinetic studies.
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| Enzyme Assay |
A typical non-cellular protocol for using N-Acetyl mesalazine-d3 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 DMSO. 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 to achieve a final concentration of 100-200 ng/mL. Proteins are precipitated by adding 150 uL of ice-cold acetonitrile containing 0.1% formic acid. The mixture is vortexed for 30 seconds 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 positive ion mode with multiple reaction monitoring (MRM) for the specific transitions of N-Acetyl mesalazine 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 N-Acetyl mesalazine-d3 as an internal standard involves the quantification of N-Acetyl mesalazine in cultured cells. Intestinal epithelial cells (e.g., Caco-2 or HT-29 cells) are seeded in 6-well plates at a density of 5×10⁵ cells/well and cultured in appropriate medium for 24-48 hours. After treatment with mesalazine (non-labeled, 0.1-10 mM) for various time periods, 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 concentrations of mesalazine and its metabolite N-Acetyl mesalazine are quantified. The internal standard corrects for extraction efficiency and matrix effects.
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| Animal Protocol |
A typical in vivo animal protocol for using N-Acetyl mesalazine-d3 as an internal standard involves the quantification of mesalazine and its metabolite N-Acetyl mesalazine in plasma and tissues. Male Sprague-Dawley rats (200-250 g) are administered mesalazine (non-labeled) via oral gavage at a dose of 50-100 mg/kg. Blood samples (200 uL) are collected via the tail vein into EDTA-coated tubes at various time points (0, 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, rats are euthanized at the final time point, and the colon and other tissues are harvested, rinsed with PBS, and homogenized in PBS (100 mg tissue/1 mL PBS). For extraction, 50 uL of plasma or tissue homogenate is mixed with 10 uL of internal standard solution (1 ug/mL) and 150 uL of acetonitrile. After centrifugation, the supernatant is analyzed by LC-MS/MS. The concentrations of mesalazine and N-Acetyl mesalazine are quantified using calibration curves prepared with the same internal standard.
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| ADME/Pharmacokinetics |
As an analytical internal standard, N-Acetyl mesalazine-d3 is not characterized by typical pharmacokinetic parameters. However, the non-labeled N-Acetyl mesalazine is the major metabolite of mesalazine (5-ASA). After oral administration of mesalazine, it is rapidly acetylated to N-Acetyl mesalazine. The plasma concentration of N-Acetyl mesalazine is typically higher than that of the parent drug due to extensive first-pass metabolism. N-Acetyl mesalazine is eliminated primarily in the urine, with a plasma elimination half-life (t1/2) of approximately 5-10 hours, which is longer than that of mesalazine (which is ~1 hour). The compound is highly bound to plasma proteins (>80%). The deuterated internal standard is used to accurately quantify N-Acetyl mesalazine in biological samples for pharmacokinetic studies.
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| Toxicity/Toxicokinetics |
Toxicity data specific to N-Acetyl mesalazine-d3 are not available. The non-labeled N-Acetyl mesalazine is a metabolite of mesalazine (5-ASA), which has a well-established safety profile for the treatment of inflammatory bowel disease. Common side effects of mesalazine include gastrointestinal disturbances (nausea, diarrhea, abdominal pain), headache, and rash. Rare but serious side effects include nephrotoxicity, pancreatitis, and blood dyscrasias. N-Acetyl mesalazine is generally considered less active than mesalazine and may have a lower risk of toxicity. However, at high concentrations, it may inhibit hTPMT and alter the metabolism of thiopurine drugs, potentially increasing their toxicity. Standard laboratory safety precautions should be followed when handling this compound. It is for research use only and should not be used in humans or animals for therapeutic or diagnostic purposes. The compound should be stored at -20degC, protected from light and moisture.
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| References | |
| Additional Infomation |
N-Acetyl mesalazine-d3 (CAS# 93968-79-7) is a stable isotope-labeled compound with a molecular weight of 198.19. The molecular formula is C9H6D3NO4, and it is also known as N-Acetyl-5-aminosalicylic acid-d3 and N-Acetyl-ASA-d3. This deuterated compound is a labelled metabolite of mesalazine, a salicylic acid derivative, and an inhibitor of recombinant human thiopurine methyltransferase (hTPMT). N-Acetyl mesalazine can scavenge free radicals, reduce DNA base hydroxylation, and ameliorate mucosal inflammation. It can be used in the study of diseases such as colitis and colon cancer. The isotopically labeled version is intended for use as an internal standard for the quantification of N-Acetyl mesalazine by LC-MS. This product is for research use only and is not approved for clinical diagnostic purposes. The compound should be stored at -20degC, protected from light and moisture.
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| Molecular Formula |
C9H6D3NO4
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| Molecular Weight |
198.19
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| Exact Mass |
198.072
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| CAS # |
93968-79-7
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| Related CAS # |
N-Acetyl mesalazine;51-59-2;N-Acetyl mesalazine-d3-1;1309935-89-4
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| PubChem CID |
46779947
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| Appearance |
White to off-white solid powder
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| Density |
1.485g/cm3
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| Boiling Point |
480.202ºC at 760 mmHg
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| Melting Point |
219-224°C
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| Flash Point |
244.217ºC
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| Index of Refraction |
1.661
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| LogP |
1.121
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
14
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| Complexity |
241
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| Defined Atom Stereocenter Count |
0
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| SMILES |
[2H]C([2H])([2H])C(=O)NC1=CC(=C(C=C1)O)C(=O)O
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| InChi Key |
GEFDRROBUCULOD-FIBGUPNXSA-N
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| InChi Code |
InChI=1S/C9H9NO4/c1-5(11)10-6-2-3-8(12)7(4-6)9(13)14/h2-4,12H,1H3,(H,10,11)(H,13,14)/i1D3
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| Chemical Name |
2-hydroxy-5-[(2,2,2-trideuterioacetyl)amino]benzoic 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 |
| 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 | 5.0457 mL | 25.2283 mL | 50.4566 mL | |
| 5 mM | 1.0091 mL | 5.0457 mL | 10.0913 mL | |
| 10 mM | 0.5046 mL | 2.5228 mL | 5.0457 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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