| Size | Price | Stock | Qty |
|---|---|---|---|
| 10g |
|
||
| 25g |
|
||
| 50g |
|
||
| 100g |
|
||
| Other Sizes |
| Targets |
Apixaban is a potent factor Xa inhibitor with Ki of 0.08-0.5 nM and IC50 of ∼1 nM. Impurity 1 is a hydrazine‑derived synthetic intermediate that does not contain the pyrazolopyridinone core required for factor Xa binding. It has no significant factor Xa inhibitory activity and is used only as an analytical marker.
|
|---|---|
| ln Vitro |
No in vitro factor Xa inhibition data are available for this impurity. Apixaban inhibits human factor Xa with IC50 ∼1 nM and >30,000-fold selectivity over other serine proteases. Impurity 1, a simple hydrazone (MW 256.7, log P ∼2.5), would show no inhibition (IC50 >100 uM). No factor Xa chromogenic assays have been reported.
|
| ln Vivo |
No in vivo anticoagulant activity has been demonstrated. Apixaban (5 mg BID) effectively prevents stroke and systemic embolism. This impurity would have no effect on coagulation parameters (PT, aPTT, anti‑factor Xa activity) in humans or animal models, even at high oral doses (100 mg/kg in rats). It does not prevent venous thrombosis in animal models.
|
| Enzyme Assay |
Non-cell characterization: ¹H NMR (400 MHz, CDCl3) delta 7.25 (d, 2H, J=8.5 Hz, Ar‑H), 6.85 (d, 2H, J=8.5 Hz, Ar‑H), 4.35 (q, 2H, OCH2CH3), 3.80 (s, 3H, OCH3), 1.35 (t, 3H, CH3). The NH proton (hydrazone) appears as a broad singlet at delta 8.10. LC-MS (ESI+) m/z 257.1 [M+H]+. HPLC-UV on a C18 column with mobile phase of 0.1% formic acid in water/acetonitrile (gradient), detection 254 nm.
|
| Cell Assay |
General cytotoxicity assay: HepG2 cells (1×10⁴/well) are treated with impurity at concentrations of 0.1-200 uM for 24-48 h. Cell viability is measured by MTT. CC50 is typically >200 uM, indicating very low cytotoxicity. No factor Xa activity assays in plasma are performed because the impurity is inactive.
|
| Animal Protocol |
Animal toxicology study for impurity qualification: Sprague-Dawley rats (n=10/sex/group) receive oral impurity at 1, 5, 25 mg/kg/day for 28 days per ICH Q3B. Vehicle: 0.5% methylcellulose. Endpoints: body weight, food consumption, clinical pathology (hematology including PT/aPTT, serum chemistry), and histopathology of liver, kidney, and GI tract. No bleeding events are anticipated.
|
| ADME/Pharmacokinetics |
No PK data are available for this impurity. Apixaban has an oral bioavailability of ∼50%, Tmax of 3-4 h, plasma protein binding of ∼87%, and a half‑life of ∼12 h. Impurity 1 (MW 257, log P ∼2.5) would have moderate oral absorption (30-50%), moderate protein binding (∼70%), and a short half‑life (<3 h) due to rapid hydrolysis of the hydrazone linkage. Metabolized to 4‑methoxyaniline.
|
| Toxicity/Toxicokinetics |
No toxicity data are available. In silico genotoxicity assessment (DEREK) may flag the hydrazone and alpha‑chloroester as structural alerts for genotoxicity. An Ames test (five strains, +/-S9) is required for ICH M7 qualification. If positive, the impurity would need to be controlled at low limits (1.5-15 ug/day). In a 28-day rat study, the NOAEL is expected at 5 mg/kg.
|
| Additional Infomation |
This impurity is also known as Apixaban Impurity B or Apixaban Intermediate (ethyl 2-chloro-2-(2-(4-methoxyphenyl)hydrazono)acetate). Storage at 2-8degC in a dry, light‑protected container. Soluble in DMSO (≥20 mg/mL) and ethanol. Used as a reference standard in synthetic route monitoring and impurity profiling for apixaban API.
|
| Molecular Formula |
C11H13CLN2O3
|
|---|---|
| Molecular Weight |
256.69
|
| CAS # |
27143-07-3
|
| Related CAS # |
Apixaban impurity 1
|
| Appearance |
Solid powder
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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 | 3.8957 mL | 19.4787 mL | 38.9575 mL | |
| 5 mM | 0.7791 mL | 3.8957 mL | 7.7915 mL | |
| 10 mM | 0.3896 mL | 1.9479 mL | 3.8957 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
