| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
PPARgamma (specific agonist), p21-activated kinase 1 (PAK1), and NF-kappaB (inhibitor). 5-ASA also inhibits the activity of osteopontin (OPN).
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| ln Vitro |
Stable heavy isotopes of hydrogen, carbon, and other elements have been incorporated into drug molecules, largely as tracers for quantitation during the drug development process. Studies involving the human use of drugs with labeled deuterium suggest that these compounds may offer some advantages when compared with their nondeuterated counterparts. Deuteration has gained attention because of its potential to affect the pharmacokinetic and metabolic profiles of drugs. Deutetrabenazine is the first deuterated drug to receive Food and Drug Administration approval. This deuterated form of the drug tetrabenazine is indicated for the treatment of chorea associated with Huntington's disease as well as tardive dyskinesia. Ongoing clinical trials suggest that a number of other deuterated compounds are being evaluated for the treatment of human diseases and not merely as research tools. [1]
In cell‑free assays, 5-ASA acts as a specific agonist for PPARgamma, with no activity on PPARalpha or PPARdelta, inducing degradation of the p65 subunit of NF-kappaB. It also inhibits PAK1 at the mRNA level, suggesting an additional PPARgamma‑independent mechanism. In HT‑29 colon carcinoma cells, 5-ASA (10‑1000 uM, 12‑96 h) inhibits cell growth in a dose‑ and time‑dependent manner, with synergistic effects when combined with nimesulide. |
| ln Vivo |
5-Aminosalicylic acid (5-ASA) has an antineoplastic effect in a xenograft tumor model. To evaluate the in vivo antineoplasic effect of 5-Aminosalicylic acid, SCID mice engrafted with HT-29 colon cancer cells are treated daily for 21 consecutive days with 5-Aminosalicylic acid at 50 mM. At the end of the treatment, a reduction of 80-86% of tumor weight and volume is observed in SCID mice receiving 5-Aminosalicylic acid compared with control mice or mice treated with GW9662 alone. The antineoplastic effect of 5-Aminosalicylic acid is already detectable after 10 days of 5-Aminosalicylic acid treatment. Similar results are obtained with mice treated with 5-Aminosalicylic acid at 5 mM. Antitumorigenic effect of 5-Aminosalicylic acid is completely abolished at 21 days by simultaneous intraperitoneal administration of GW9662. Thus, the observed antineoplastic effect of 5-Aminosalicylic acid is at least partially dependent on PPARγ[4].
In vivo, in a SCID mouse xenograft model bearing HT‑29 colon cancer cells, treatment with 5-ASA (50 mM, 21 days) resulted in an 80‑86% reduction in tumor weight and volume. 5-Aminosalicylic acid-d3 disodium, as an internal standard, does not have independent in vivo activity. |
| Enzyme Assay |
General cell‑free protocol for assessing PPARgamma agonist activity: A fluorescence polarization (FP) competitive binding assay is used. Recombinant human PPARgamma ligand‑binding domain (LBD) protein (5 nM) is incubated with a fluorescently labeled PPARgamma ligand (Fluormone PLM Red, 1 nM) in a buffer (100 mM potassium phosphate, pH 7.4, 100 ug/mL BSA) at room temperature for 2 hours. Varying concentrations of 5-ASA (0.01‑1000 uM) are added. The FP signal is measured, and the IC₅0 is calculated. For the PPARgamma‑p65 degradation assay, purified PPARgamma and p65 proteins are incubated with 5-ASA, and p65 degradation is monitored by Western blot.
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| Cell Assay |
General cellular protocol for 5-ASA: HT‑29 colon carcinoma cells are seeded in 96‑well plates at 5,000 cells/well. After 24 hours, cells are treated with various concentrations of 5-ASA (10, 100, 500, 1000 uM) for 24, 48, 72, and 96 hours. Cell viability is assessed using the MTT assay. For combination studies, cells are treated with 5-ASA and nimesulide simultaneously. For mechanistic studies, cells are treated with 5-ASA (100‑1000 uM) for 24‑48 hours. Total RNA is extracted, and PAK1 mRNA levels are measured by qRT‑PCR. Protein lysates are subjected to Western blot with anti‑p65 and anti‑PPARgamma antibodies.
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| Animal Protocol |
General animal protocol for 5-ASA: SCID mice are engrafted subcutaneously with HT‑29 colon cancer cells (5×10⁶ cells). When tumors reach ~100 mm3, the mice are randomized into treatment groups (n=6/group). 5-ASA is formulated in a suitable vehicle (e.g., saline or 0.5% methylcellulose) and administered via oral gavage at doses of 100, 500, and 1000 mg/kg daily for 21 days. Tumor volume is measured twice weekly. At the end of the study, tumors are excised and weighed, and tumor tissues are processed for histology and Western blot analysis.
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| ADME/Pharmacokinetics |
General PK protocol for 5-ASA using its labeled internal standard: A sensitive and specific LC‑MS/MS method is used to quantify 5-ASA in pharmacokinetic studies. The internal standard (5-Aminosalicylic acid-d3 disodium) is added to plasma samples at a constant concentration (e.g., 100 ng/mL). The samples are processed by protein precipitation with acetonitrile. The m/z transitions for 5-ASA (e.g., 154→110) and for the d3 internal standard (157→113) are monitored. Pharmacokinetic parameters (Cmax, Tmax, AUC, t½, CL, Vd) are calculated from the concentration‑time data.
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| Toxicity/Toxicokinetics |
General toxicity protocol for 5-ASA: A 90‑day repeat‑dose oral toxicity study is performed in rats at doses of 100, 300, and 1000 mg/kg/day. Parameters include clinical signs, body weight, food consumption, hematology (CBC, differential), serum chemistry (ALT, AST, BUN, creatinine, electrolytes), urinalysis (pH, specific gravity, protein, blood), and histopathology of major organs. The primary adverse effects of 5-ASA are gastrointestinal (diarrhea, nausea) and renal (interstitial nephritis).
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| References |
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| Additional Infomation |
5-Aminosalicylic acid-d3 disodium has the molecular formula C₇H2D3NNa2O3 and a molecular weight of 200.12 g/mol. The unlabeled 5-ASA (mesalamine) is a first‑line treatment for ulcerative colitis and Crohn‘s disease. Its mechanism of action in IBD is primarily through local mucosal effects: activation of PPARgamma in colonic epithelial cells inhibits NF-kappaB signaling, reducing the production of pro‑inflammatory cytokines (e.g., IL‑1, IL‑6, TNF‑alpha). It also inhibits the activity of osteopontin (OPN), a cytokine involved in inflammatory and fibrotic processes. The deuterated compound is used as an internal standard for LC‑MS quantification.
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| Molecular Formula |
C7H2D3NNA2O3
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|---|---|
| Molecular Weight |
200.12
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| Appearance |
Typically exists as solid at room temperature
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| Synonyms |
5-Aminosalicylic acid disodium salt-d3; Mesalamine-d3 disodium; 5-ASA-d3 disodium; Mesalazine-d3 disodium
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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 | 4.9970 mL | 24.9850 mL | 49.9700 mL | |
| 5 mM | 0.9994 mL | 4.9970 mL | 9.9940 mL | |
| 10 mM | 0.4997 mL | 2.4985 mL | 4.9970 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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