| Size | Price | Stock | Qty |
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| 5mg |
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| Other Sizes |
| Targets |
Apixaban-d3 is a stable isotope-labeled internal standard. Its unlabeled parent compound, Apixaban, is a highly potent, selective, reversible, and orally bioavailable inhibitor of activated factor X (FXa). The primary target of Apixaban is factor Xa (Ki = 0.08 nM for both free and prothrombinase-bound FXa), a key serine protease in the coagulation cascade. By directly inhibiting FXa, Apixaban blocks the conversion of prothrombin to thrombin, thereby reducing the generation of fibrin and the formation of thrombus (clot). Apixaban does not require antithrombin III for its activity and does not affect platelet aggregation directly. The deuterated version (Apixaban-d3) mimics this same binding selectivity but is used for analytical quantification in research and quality control applications, not for therapeutic purposes.
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| ln Vitro |
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as tracers that influence measurement during the drug development process. It's possible that the pharmacokinetics and functional range of medications contribute to the concern over mutagenesis [1].
The in vitro biological activity of Apixaban-d3 is presumed to be identical to its unlabeled parent, Apixaban. In vitro assays have demonstrated that Apixaban is a highly potent and selective direct inhibitor of human factor Xa. Using a purified enzyme assay (chromogenic substrate S-2765), Apixaban inhibits FXa with a Ki value of 0.08 nM, and it is at least 10,000-fold more selective for FXa than for other serine proteases (thrombin, trypsin, chymotrypsin). In human plasma clotting assays, Apixaban doubles the prothrombin time (PT) at a concentration of 0.4-0.6 uM. In thrombin generation assays using human platelet-poor plasma, Apixaban (1-100 nM) prolongs the lag time and reduces the peak thrombin generation in a concentration-dependent manner. The labeled Apixaban-d3 is used as an internal standard in LC-MS to accurately quantify drug concentrations in these in vitro assay systems. |
| ln Vivo |
The in vivo activity of Apixaban-d3 is not directly evaluated, but its unlabeled parent is a well-characterized anticoagulant. In a rabbit model of venous thrombosis, oral administration of Apixaban (0.1-3 mg/kg) produced dose-dependent antithrombotic effects, reducing thrombus weight by up to 90% at 3 mg/kg. In rat models of arterial thrombosis, Apixaban (0.03-0.3 mg/kg i.v. or 1-10 mg/kg p.o.) reduced thrombus formation without significantly increasing bleeding time at therapeutic doses. In dogs, Apixaban (0.5-5 mg/kg p.o.) produced dose-dependent prolongation of clotting times (PT, aPTT) and inhibition of FXa activity. The antithrombotic efficacy of Apixaban correlates with plasma drug levels, with an EC₅0 for thrombus reduction of approximately 50-100 ng/mL. Apixaban-d3 is used as an internal standard for LC-MS to accurately quantify drug levels in plasma and tissues, enabling precise PK/PD (pharmacokinetic/pharmacodynamic) modeling of the dose-response relationship.
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| Enzyme Assay |
A generic non-cell-based assay for Apixaban-d3 involves its use as an internal standard in an LC-MS/MS method for quantifying Apixaban in human plasma. Prepare a standard stock solution of unlabeled Apixaban in DMSO (1 mg/mL). Prepare a separate stock solution of Apixaban-d3 at the same concentration (or use commercially prepared solution). Prepare calibration standards by spiking the unlabeled analyte into blank human plasma to achieve concentrations ranging from 0.5 to 500 ng/mL. Add a fixed concentration of the internal standard (e.g., 50 ng/mL) to each calibration standard. Also prepare blank and double-blank samples. For sample preparation, add 200 uL of acetonitrile containing 0.1% formic acid to 100 uL of plasma to precipitate proteins. Vortex and centrifuge at 12,000g for 10 minutes. Transfer the supernatant to an autosampler vial. Analyze by LC-MS/MS in positive ESI mode. Monitor the mass transitions: m/z 460.2 → 443.2 (loss of NH3) for Apixaban, and m/z 463.2 → 446.2 for Apixaban-d3 (13C-d3 labeled variant). Construct the calibration curve by plotting the peak area ratio (analyte/IS) vs. the nominal concentration. Method sensitivity should achieve a lower limit of quantification (LLOQ) of 0.5 ng/mL or better.
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| Cell Assay |
A standard in vitro cell-based protocol for the unlabeled Apixaban is not typical, as Apixaban acts on circulating coagulation factors. However, its anticoagulant effects can be assessed in human whole blood or platelet-rich plasma. Collect whole blood from healthy human volunteers into sodium citrate tubes (3.2% citrate). For the thrombin generation assay (TGA), prepare platelet-poor plasma (PPP) by centrifugation at 2,000g for 15 minutes. To 80 uL of PPP, add increasing concentrations of unlabeled Apixaban (0, 10, 30, 100, 300, 1000 ng/mL) and 20 uL of PPP-Reagent (containing low amounts of tissue factor and phospholipids). Initiate the reaction by adding 20 uL of FluCa substrate (containing calcium and a fluorogenic thrombin substrate). Monitor fluorescence (excitation 360 nm, emission 460 nm) continuously for 60-120 minutes at 37degC using a fluorometer plate reader. Calculate the following thrombin generation parameters: lag time (minutes), peak thrombin (nM), time to peak (minutes), and endogenous thrombin potential (ETP, nM·min). Determine the IC₅0 for each parameter. Use Apixaban-d3 as an internal standard in an LC-MS analysis of the PPP samples to validate the drug concentrations.
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| Animal Protocol |
A typical in vivo animal protocol for Apixaban-d3 involves a pharmacokinetic (PK) and pharmacodynamic (PD) study in dogs or rats. For a rat study, use male Sprague-Dawley rats (250-300 g, n = 6 per group). Administer a single oral dose of unlabeled Apixaban (5 mg/kg) suspended in 0.5% methylcellulose (with 0.02% Tween 80). Collect blood samples via tail vein at various time points (0, 0.5, 1, 2, 4, 6, 8, 12, 24, 36, 48 h) into 3.2% sodium citrate tubes (9:1 ratio). Centrifuge the blood at 2,000g for 15 minutes to obtain platelet-poor plasma. Store plasma samples at -80degC. For PK analysis, spike plasma samples (50 uL) with a fixed amount of Apixaban-d3 as internal standard. Precipitate proteins with acetonitrile, centrifuge, and analyze the supernatant by LC-MS/MS. For PD analysis, measure the anti-FXa activity in plasma using a chromogenic anti-Xa assay kit (e.g., Rotachrom). Plot the relationship between plasma Apixaban concentration and anti-FXa activity to generate PK/PD correlation curves. Calculate PK parameters including Cmax, Tmax, AUC(0-48), t½, and oral clearance using non-compartmental analysis.
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| ADME/Pharmacokinetics |
Apixaban-d3 is an analytical internal standard, so its PK is identical to its unlabeled parent, Apixaban. Apixaban is an oral direct factor Xa inhibitor approved for the prevention and treatment of thromboembolic disorders. In humans, Apixaban is rapidly absorbed after oral administration, with Tmax of 1-4 hours and absolute oral bioavailability of approximately 50-60% (for the 5 mg dose). It exhibits linear pharmacokinetics over the therapeutic dose range (2.5-10 mg). Apixaban is highly plasma protein-bound (approximately 87%), primarily to albumin. The volume of distribution is approximately 21 L (moderate distribution). Apixaban is metabolized primarily by CYP3A4/5 (with minor contributions from CYP1A2, 2C8, 2C9, 2C19, and 2J2), with O-demethylation and hydroxylation being the major pathways. The majority of the dose (~75%) is excreted in feces as unchanged drug and metabolites, while ~25% is excreted in urine. The terminal elimination half-life is approximately 12-15 hours in healthy subjects, allowing once or twice daily dosing. The labeled Apixaban-d3 is critical for accurate quantification of the drug in plasma and tissues for PK studies.
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| Toxicity/Toxicokinetics |
Apixaban-d3 is a research-grade internal standard, not a therapeutic agent. Its unlabeled parent, Apixaban, is an approved anticoagulant with a well-characterized safety profile. The most common adverse effects in clinical use are bleeding (major bleeding occurs in approximately 2-3% of patients per year) and gastrointestinal symptoms (nausea, dyspepsia). Apixaban is contraindicated in patients with active pathological bleeding, severe hepatic impairment, or mechanical heart valves. The oral LD₅0 of Apixaban in rodents is >5,000 mg/kg, indicating low acute toxicity. In chronic toxicology studies in rats and dogs (up to 6 months), no target organ toxicity was observed at exposures up to 20-30 times the human therapeutic exposure. Apixaban is not genotoxic (Ames test, chromosomal aberration assay negative) and is not carcinogenic in long-term rodent studies. For laboratory handling, standard precautions for anticoagulant research compounds should be used. Apixaban-d3 should be stored as a powder at -20degC, protected from light and moisture. Solutions in DMSO can be stored at -80degC for up to 6 months.
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| References | |
| Additional Infomation |
Apixaban-d3 (Apixaban-13C-d3) is the stable isotope-labeled version of Apixaban, a direct and selective inhibitor of activated factor X (FXa). Apixaban is an orally bioavailable anticoagulant that was approved by the FDA in 2012 for the prevention of stroke and systemic embolism in patients with non-valvular atrial fibrillation, and subsequently for the treatment of deep vein thrombosis (DVT) and pulmonary embolism (PE), as well as for the prophylaxis of DVT following hip or knee replacement surgery. Apixaban was developed by Bristol-Myers Squibb and Pfizer and is marketed under the brand name Eliquis. Its high selectivity for FXa (Ki = 0.08 nM) and favorable pharmacokinetic profile have made it a leading drug in the anticoagulant market. Apixaban-d3 serves as a critical internal standard for LC-MS/MS bioanalysis in research applications, including bioequivalence studies, drug-drug interaction studies, and therapeutic drug monitoring research. For research use only, not for human diagnostic or therapeutic applications.
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| Molecular Formula |
C25H22D3N5O4
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|---|---|
| Molecular Weight |
462.52
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| Exact Mass |
462.209
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| CAS # |
1131996-12-7
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| Related CAS # |
Apixaban;503612-47-3
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| PubChem CID |
25226058
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| Appearance |
Off-white to light yellow solid powder
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| LogP |
3.713
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
5
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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 |
[2H]C([2H])([2H])OC1=CC=C(C=C1)N2C3=C(CCN(C3=O)C4=CC=C(C=C4)N5CCCCC5=O)C(=N2)C(=O)N
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| InChi Key |
QNZCBYKSOIHPEH-FIBGUPNXSA-N
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| InChi Code |
InChI=1S/C25H25N5O4/c1-34-19-11-9-18(10-12-19)30-23-20(22(27-30)24(26)32)13-15-29(25(23)33)17-7-5-16(6-8-17)28-14-3-2-4-21(28)31/h5-12H,2-4,13-15H2,1H3,(H2,26,32)/i1D3
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| Chemical Name |
7-oxo-6-[4-(2-oxopiperidin-1-yl)phenyl]-1-[4-(trideuteriomethoxy)phenyl]-4,5-dihydropyrazolo[3,4-c]pyridine-3-carboxamide
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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 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.) |
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| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1621 mL | 10.8103 mL | 21.6207 mL | |
| 5 mM | 0.4324 mL | 2.1621 mL | 4.3241 mL | |
| 10 mM | 0.2162 mL | 1.0810 mL | 2.1621 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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