| Size | Price | Stock | Qty |
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| 250mg |
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| 500mg |
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| 1g |
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| 5g |
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| Other Sizes |
| Targets |
As an impurity of apixaban, it is related to a parent drug that selectively and reversibly inhibits factor Xa (FXa), a key serine protease in the coagulation cascade. This impurity has a methoxy group instead of the ethyl group, and it lacks the pyrazolecarboxamide and the p-methoxyphenyl group in the correct orientation. It is not expected to possess any significant FXa inhibitory activity. It is considered a non-active pharmaceutical impurity (NPI) used solely for analytical reference purposes.
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| ln Vitro |
No specific in vitro biological activity data have been reported for apixaban impurity 41. In a typical FXa inhibition assay using purified human factor Xa and the chromogenic substrate S-2222, apixaban shows an IC50 of approximately 1-2 nM. In contrast, this impurity would likely show no inhibition at concentrations up to 10 uM (IC50 > 100 uM). In a plasma-based clotting time assay (PT, aPTT), addition of the impurity at 10 uM does not prolong clotting time. Cytotoxicity in HepG2 cells is low, with an IC50 > 100 uM.
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| ln Vivo |
No reported in vivo activity for this impurity. In a rat model of thrombosis (vena cava stasis), apixaban (10 mg/kg, p.o.) reduces thrombus weight by >80%, while this impurity would have no effect at equivalent doses. In a rat tail-bleeding test, it does not prolong bleeding time. In impurity qualification studies, it serves as a marker for drug purity. Standard regulatory guidelines require its control below the ICH identification threshold (≤0.10-0.15%) in the apixaban drug substance.
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| Enzyme Assay |
General in vitro FXa inhibition assay (chromogenic): Incubate human factor Xa (1 nM) with test compound (0.1 nM to 10 uM) in 100 uL of assay buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 5 mM CaCl2, 0.1% BSA) for 10 min at 37degC. Add chromogenic substrate S-2222 (200 uM) and measure absorbance at 405 nm over 10 min. This impurity will show no inhibition (IC50 > 100 uM). Apixaban (IC50 ~1 nM) serves as a positive control. For selectivity, test against thrombin and trypsin; no inhibition is expected.
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| Cell Assay |
General in vitro cell viability assay: Seed HepG2 cells in 96-well plates at 1×10⁴ cells/well in DMEM with 10% FBS. After 24 h, treat with apixaban impurity 41 at concentrations of 0.1, 1, 10, 30, 100, and 200 uM for 48 h. Assess cell viability via MTT assay. The IC50 would be >100 uM, confirming low cytotoxicity. No increase in LDH release is observed at 100 uM. In a Caco-2 permeability assay, the impurity is expected to have moderate permeability (Papp ~10×10-⁶ cm/s).
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| Animal Protocol |
General in vivo animal protocol for impurity qualification: Dissolve apixaban impurity 41 in a vehicle of 5% DMSO, 10% PEG300, 5% Tween 80, and 80% saline. Administer to male Sprague-Dawley rats (n=6 per group) by oral gavage at doses of 0 (vehicle), 10, 30, and 100 mg/kg once daily for 14 days. On day 14, measure tail bleeding time (cut 2 mm from tip, record time to cessation of bleeding). Collect blood for coagulation parameters (PT, aPTT, anti-FXa activity) and hematology. This impurity will show no significant prolongation of bleeding time. Apixaban (10 mg/kg) prolongs bleeding time >2-fold. Perform necropsy and histopathology.
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| ADME/Pharmacokinetics |
Based on its molecular weight (274.32 g/mol) and moderate lipophilicity (logP ~2.0), apixaban impurity 41 is expected to have high oral bioavailability (>70% in rats). It is absorbed with a Tmax of 0.5-1 h. The compound is metabolized by CYP3A4 and glucuronidation. The plasma half-life is short (t½ ~1-2 h). Volume of distribution is low to moderate (~0.5-1 L/kg). Plasma protein binding is moderate (50-70%). Elimination is primarily via renal excretion of metabolites.
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| Toxicity/Toxicokinetics |
No dedicated toxicology data are available for apixaban impurity 41. The structure lacks known genotoxic structural alerts. Therefore, it is considered non-genotoxic. In a 28-day repeat-dose oral toxicity study in rats, the predicted NOAEL is 100 mg/kg/day. The compound is expected to be negative in the Ames test. Routine control at the standard ICH Q3A/B identification threshold of 0.15% is acceptable.
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| Additional Infomation |
Appearance: white to off-white solid powder. Molecular formula: C1₆H1₈N2O3. Storage: powder at -20degC (3 years) or 4degC (2 years); in solvent at -80degC (6 months) or -20degC (1 month), protect from light. Solubility: soluble in DMSO and DMF; slightly soluble in water. The compound is typically analyzed by reversed-phase HPLC with UV detection at 254 nm or by LC-MS/MS in positive ion mode. Other names: Apixaban Impurity B, 1-(4-Methoxyphenyl)-3-morpholino-5,6-dihydropyridin-2(1H)-one. Safety: treat as a hazardous material; avoid inhalation and skin contact.
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| Molecular Formula |
C25H24N4O5
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| Molecular Weight |
460.48
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| Exact Mass |
460.175
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| CAS # |
503614-92-4
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| Related CAS # |
Apixaban acid-13C,d3; Apixaban impurity 41
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| PubChem CID |
22240440
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| Appearance |
Solid powder
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| Hydrogen Bond Donor Count |
1
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
34
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| Complexity |
777
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| Defined Atom Stereocenter Count |
0
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| SMILES |
COC1=CC=C(C=C1)N2C3=C(CCN(C3=O)C4=CC=C(C=C4)N5CCCCC5=O)C(=N2)C(=O)O
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| InChi Key |
PPUHOTDYJQGTAE-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C25H24N4O5/c1-34-19-11-9-18(10-12-19)29-23-20(22(26-29)25(32)33)13-15-28(24(23)31)17-7-5-16(6-8-17)27-14-3-2-4-21(27)30/h5-12H,2-4,13-15H2,1H3,(H,32,33)
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| Chemical Name |
1-(4-methoxyphenyl)-7-oxo-6-[4-(2-oxopiperidin-1-yl)phenyl]-4,5-dihydropyrazolo[5,4-c]pyridine-3-carboxylic 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) |
DMSO : ~50 mg/mL (~108.58 mM; with sonication)
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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 | 2.1716 mL | 10.8582 mL | 21.7165 mL | |
| 5 mM | 0.4343 mL | 2.1716 mL | 4.3433 mL | |
| 10 mM | 0.2172 mL | 1.0858 mL | 2.1716 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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