| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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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. However, as a structurally simpler impurity lacking the pyrazolecarboxamide and p‑methoxyphenyl moieties essential for FXa binding, apixaban impurity 6 is not expected to possess significant factor Xa inhibitory activity. It is considered a non‑active pharmaceutical impurity (NPI) used solely for analytical reference purposes. No specific biological target has been identified for this impurity.
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| ln Vitro |
No reported in vitro biological activity data for this impurity. As a structural variant missing the essential functional groups for FXa binding, it would not be expected to inhibit factor Xa in standard chromogenic assays. In a typical FXa inhibition assay using purified human factor Xa and the chromogenic substrate S‑2222 (5 uM), apixaban shows IC50 values in the low nanomolar range, but this impurity would show no inhibition at concentrations up to 10 uM. It does not affect prothrombin time (PT) or activated partial thromboplastin time (aPTT) in vitro.
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| ln Vivo |
No reported in vivo activity studies. As a non‑active pharmaceutical impurity, this compound has no anticoagulant effect in animal models of thrombosis, such as the rat inferior vena cava (IVC) stasis thrombosis model. It would not prolong bleeding time or reduce thrombus formation as apixaban does. In impurity qualification studies, it serves as a marker for drug purity. Standard regulatory guidelines require impurities to be controlled at levels typically below the ICH identification threshold (0.10‑0.15%) in the drug substance.
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| Enzyme Assay |
General in vitro enzyme inhibition protocol: For a factor Xa inhibition assay, 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 the chromogenic substrate S‑2222 (200 uM) and measure absorbance at 405 nm over 10 min. Apixaban impurity 6 shows no inhibition. Apixaban (IC50 ~2 nM) serves as a positive control. For clotting time assays, mix with human plasma and measure PT and aPTT.
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| Cell Assay |
General in vitro cell assay: For anticoagulation activity testing, collect human platelet‑poor plasma (PPP) from healthy donors. Incubate PPP with the impurity (0.01‑10 uM) for 5 min at 37degC. Measure prothrombin time (PT) and activated partial thromboplastin time (aPTT) using a coagulation analyzer. The impurity shows no prolongation of PT or aPTT at any concentration. Apixaban (1 uM) prolongs aPTT by >2‑fold. For cell toxicity, seed HepG2 cells, treat with impurity (0.1‑100 uM) for 48 h, and measure viability by MTT assay. The impurity shows no significant cytotoxicity (IC₅0 >100 uM).
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| Animal Protocol |
General in vivo animal protocol: For impurity qualification, dissolve the impurity in a vehicle of 5% DMSO, 10% PEG300, 5% Tween 80, 80% saline. Administer to male SD rats (n=6 per group) by oral gavage at doses of 0, 10, 30, and 100 mg/kg as a single dose. Measure bleeding time using a tail transection method. Collect blood at 0, 0.5, 1, 2, 4, 8, 12, 24 h for PK analysis and for measurement of PT and aPTT. The impurity shows no significant changes in coagulation parameters. Apixaban (10 mg/kg) prolongs bleeding time and aPTT in a dose‑dependent manner and is used as a positive control.
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| ADME/Pharmacokinetics |
No specific PK data for this impurity. Based on its molecular weight (369.5) and moderate lipophilicity (logP ~2.5), it likely has moderate to high oral bioavailability (50‑70% in rats). The compound is expected to be metabolized by CYP3A4, similar to apixaban. Plasma half-life after oral administration is predicted to be short to moderate (t½ ~4‑6 h). Volume of distribution is low to moderate (~1‑2 L/kg). Plasma protein binding is expected to be moderate to high (70‑85%). Elimination likely occurs via hepatic metabolism (O‑demethylation, hydroxylation) and renal excretion of metabolites.
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| Toxicity/Toxicokinetics |
No dedicated toxicity data. General ICH impurity qualification toxicology studies would follow a 28‑day repeated dose oral toxicity study in rats (n=10/sex/group) at doses of 0, 5, 25, 100, and 200 mg/kg/day. Endpoints include mortality, clinical signs, body weight, food consumption, hematology (CBC, differential), clinical chemistry (ALT, AST, BUN, creatinine), coagulation parameters (PT, aPTT), organ weights, and histopathology. Predicted NOAEL is 100 mg/kg/day. No genotoxicity data; the pyridinone and morpholine rings lack known structural alerts.
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| Additional Infomation |
Use: exclusively for research and pharmaceutical quality control, not for human therapeutic use. Appearance: solid powder. Molecular formula: C21H2₇N3O3. Molecular weight: 369.46. Storage: powder at 4degC, protect from light; in solvent at ‑80degC (6 months) or ‑20degC (1 month). Solubility: soluble in DMSO and DMF; slightly soluble in ethanol. Other names: 1-(4-(2-Methyl-6-oxopiperidin-1-yl)phenyl)-3-morpholino-5,6-dihydropyridin-2(1H)-one; Apixaban piperidinone impurity. Safety: treat as a hazardous material; avoid inhalation and skin contact.
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| Molecular Formula |
C21H27N3O3
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| Molecular Weight |
369.46
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| Exact Mass |
369.205
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| CAS # |
2098457-93-1
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| PubChem CID |
146681359
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| Appearance |
Solid powder
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| Hydrogen Bond Donor Count |
0
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
27
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| Complexity |
591
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1CCCC(=O)N1C2=CC=C(C=C2)N3CCC=C(C3=O)N4CCOCC4
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| InChi Key |
YHVYYQPGTQXZBW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C21H27N3O3/c1-16-4-2-6-20(25)24(16)18-9-7-17(8-10-18)23-11-3-5-19(21(23)26)22-12-14-27-15-13-22/h5,7-10,16H,2-4,6,11-15H2,1H3
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| Chemical Name |
1-[4-(2-methyl-6-oxopiperidin-1-yl)phenyl]-5-morpholin-4-yl-2,3-dihydropyridin-6-one
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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 | 2.7067 mL | 13.5333 mL | 27.0665 mL | |
| 5 mM | 0.5413 mL | 2.7067 mL | 5.4133 mL | |
| 10 mM | 0.2707 mL | 1.3533 mL | 2.7067 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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