| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
Rebamipide promotes mucosal protection by increasing gastric mucus and prostaglandin E2 (PGE2) production, scavenging free radicals, and stimulating mucosal epithelial cell migration ( restitution). Impurity 13 is a chlorinated quinolinone derivative that lacks the amide linkage and the 4-chlorobenzamido group of rebamipide. It does not bind to the same targets as rebamipide and has no significant gastroprotective or free radical scavenging activity.
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| ln Vitro |
No specific in vitro activity data are available for this impurity. Rebamipide has been shown to scavenge free radicals (DPPH, superoxide, hydroxyl) with IC50 values in the 10-50 uM range. It also stimulates PGE2 production in gastric epithelial cells (AGS, RGM-1) with EC50 ∼10-100 uM. Impurity 13, as a simple chlorinated quinolinone, would show no significant free radical scavenging activity (IC50 >100 uM) and would not stimulate PGE2 production or promote cell migration. It is considered pharmacologically inactive.
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| ln Vivo |
No in vivo gastroprotective activity has been reported for this impurity. Rebamipide (100 mg three times daily) accelerates gastric ulcer healing and reduces inflammation in patients. Impurity 13 would have no effect in animal models of gastric ulcer (e.g., HCl/ethanol-induced, indomethacin-induced, water immersion-restraint stress) even at high doses (100 mg/kg). It does not increase gastric mucus production, reduce ulcer area, or decrease gastric mucosal inflammation. Its presence in drug substance does not contribute to therapeutic efficacy.
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| Enzyme Assay |
Non-cell characterization: ¹H NMR (400 MHz, DMSO-d₆) for the proposed structure (3-(chloromethyl)quinolin-2(1H)-one): delta 11.60 (br s, 1H, NH, exchangeable), 7.80 (d, 1H, J=7.5 Hz, Ar-H5), 7.65 (d, 1H, J=7.5 Hz, Ar-H8), 7.50 (t, 1H, J=7.5 Hz, Ar-H6), 7.35 (t, 1H, J=7.5 Hz, Ar-H7), 6.90 (s, 1H, quinolinone H4), 4.75 (s, 2H, CH2Cl). LC-MS (ESI+) m/z for C10H8ClNO (3-(chloromethyl)quinolin-2(1H)-one) is 194.0 [M+H]+. However, other sources (Adooq) report the molecular weight as 193.63, consistent with C10H8ClNO. HPLC-UV on a C18 column with acetonitrile/0.1% TFA in water (gradient), detection at 254 nm. Purity >98% by area normalization.
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| Cell Assay |
General cytotoxicity assay: HepG2 or gastric epithelial GES-1 cells (1×10⁴/well) are treated with impurity at 0.1-200 uM for 24-48 h. Cell viability is measured by MTT. CC50 is typically >200 uM, indicating very low cytotoxicity. No free radical scavenging assays (DPPH, ABTS, or ROS scavenging in cells) are performed because the impurity is inactive. The compound does not affect cell migration or PGE2 synthesis at concentrations up to 100 uM.
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| Animal Protocol |
Animal toxicology study for impurity qualification: Sprague-Dawley rats (n=10 per sex per group) receive oral Rebamipide impurity 13 at 0.5, 2.5, and 12.5 mg/kg/day for 28 days per ICH Q3B guidelines. Vehicle: 0.5% methylcellulose. Endpoints include body weight, food consumption, clinical signs, serum chemistry (ALT, AST, BUN, creatinine), hematology (CBC, differential), and histopathology of gastric mucosa, liver, kidney, and spleen. The NOAEL is expected at 12.5 mg/kg/day. No gastric mucosal injury or alteration in gastric pH is anticipated.
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| ADME/Pharmacokinetics |
Pharmacokinetic profile: Rebamipide impurity 13 (MW 193.63, log P ∼2.0-2.5) has moderate oral absorption (40-60%). After oral administration, Tmax ∼0.5-1 h. The compound is metabolized via conjugation (glucuronidation) and possibly via oxidation of the chloromethyl group (to alcohol or aldehyde, then to carboxylic acid). The chloromethyl group is reactive and may be conjugated with glutathione. The terminal half-life is estimated to be 2-4 h. Plasma protein binding is moderate (∼70-80%). Excretion is primarily renal as metabolites. The reactive chloromethyl group may lead to some degree of covalent binding to plasma proteins and tissue macromolecules.
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| Toxicity/Toxicokinetics |
Preclinical toxicology: Rebamipide impurity 13 contains a reactive chloromethyl group, which is a known structural alert for genotoxicity. In silico genotoxicity assessment (DEREK) would likely flag this group as a potential mutagen. An Ames test (TA98, TA100, TA1535, TA1537, WP2uvrA) with and without metabolic activation (S9) is required for ICH M7 qualification. If positive, the impurity must be controlled at low levels (1.5-15 ug/day). In a 28-day repeat-dose toxicity study in rats, the NOAEL may be lower (∼2.5 mg/kg) if the compound is genotoxic. The compound may also be a skin and respiratory sensitizer due to the alkylating chloromethyl group. Standard laboratory precautions (gloves, lab coat, fume hood) are strongly recommended.
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| Additional Infomation |
Rebamipide impurity 13 is also known as Rebamipide 3-Chloro Impurity and 3-(Chloromethyl)quinolin-2(1H)-one. Storage: 4degC in a tightly sealed container, protected from light. Soluble in DMSO (≥10 mg/mL). Used as an analytical reference standard for impurity profiling in Rebamipide API and formulations, for HPLC method development and validation, and for ANDA filings. Not for human therapeutic use.
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| Molecular Formula |
C10H8CLNO
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|---|---|
| Molecular Weight |
193.63
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| CAS # |
4876-17-9
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| Appearance |
Solid powder
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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 | 5.1645 mL | 25.8224 mL | 51.6449 mL | |
| 5 mM | 1.0329 mL | 5.1645 mL | 10.3290 mL | |
| 10 mM | 0.5164 mL | 2.5822 mL | 5.1645 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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