| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Rivaroxaban potently inhibits human factor Xa with Ki of 0.4 nM and IC50 of 0.7 nM in cell-free assays. Impurity 3 has a molecular weight of 608.60 and contains the factor Xa pharmacophore but in a dimeric form that likely prevents binding. It probably shows no measurable factor Xa inhibitory activity. For analytical purposes, its target is not relevant, as it serves only as a reference standard.
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|---|---|
| ln Vitro |
No in vitro activity data are available for this impurity. Rivaroxaban selectively inhibits human factor Xa with >10,000-fold selectivity over other serine proteases. This impurity, as a dimeric derivative, lacks the proper 3D conformation to access the factor Xa active site. Even at 100 uM, no inhibitory activity against factor Xa is expected.
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| ln Vivo |
No in vivo activity data exist. Rivaroxaban has excellent oral bioavailability (~80-100%) with Tmax 2-4 h, t½ 5-9 h, and >90% plasma protein binding. This impurity has a significantly larger MW (608.60), making oral absorption unlikely. Even if absorbed, it would not inhibit factor Xa in vivo and would not affect coagulation parameters.
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| Enzyme Assay |
Structural confirmation of Rivaroxaban impurity 3 uses ¹H NMR (400 MHz, DMSO-d₆) showing characteristic urea NH protons at delta 6.0-6.5 ppm, oxazolidinone protons at delta 4.0-4.8 ppm, and morpholinone ring protons. ¹3C NMR confirms the urea carbonyl. LC-MS (ESI+) detects [M+H]+ at m/z 609.2. HPLC-UV on a C18 column (150×4.6 mm, 3.5 um) with mobile phase of 0.05 M KH2PO4 (pH 3.0)/acetonitrile (gradient 20→80% B), detection 254 nm, determines purity (>95%).
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| Cell Assay |
General cytotoxicity assay: HEK293 or HepG2 cells (1×10⁴/well) are treated with impurity at 0.1-100 uM for 24-48 h. Cell viability measured by MTT or CellTiter-Glo. No specific factor Xa inhibition assays in cells are performed since the impurity is pharmacologically inactive. CC50 is typically >100 uM, indicating low cytotoxicity.
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| Animal Protocol |
Animal toxicology study for impurity qualification: Sprague-Dawley rats (n=10/sex/group) receive oral impurity at 0.5, 2.5, 12.5 mg/kg/day for 28 days per ICH Q3B. Vehicle: 0.5% methylcellulose. Endpoints include body weight, clinical signs, hematology (including PT, aPTT, factor Xa activity), serum chemistry, and histopathology of liver, kidney, and GI tract.
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| ADME/Pharmacokinetics |
No PK data are available for this impurity. For Rivaroxaban, oral bioavailability is 80-100%, Tmax 2-4 h, plasma protein binding 92-95%, t½ 5-9 h. Impurity 3 has a larger MW (608.60) and likely much lower oral absorption (<10%). It probably shows higher plasma protein binding (>99%) due to its lipophilicity and would be excreted slowly with t½ >24 h.
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| Toxicity/Toxicokinetics |
No toxicity data are available. In silico genotoxicity assessment (DEREK, Sarah Nexus) per ICH M7 should be performed to identify structural alerts. An Ames test (TA98, TA100, TA1535, TA1537, WP2uvrA) with and without metabolic activation is required for qualification. In a 28-day rat study, the NOAEL is expected at 2.5 mg/kg. No specific organ toxicity is anticipated.
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| Additional Infomation |
This impurity is also known as Rivaroxaban Impurity D. Storage at 2-8degC, protected from light. Soluble in DMSO (≥10 mg/mL) and acetonitrile. It is a critical marker for stability-indicating HPLC methods and is listed in various pharmacopoeial monographs. Used for ANDA filings and method validation.
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| Molecular Formula |
C29H32N6O9
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|---|---|
| Molecular Weight |
608.60
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| Exact Mass |
608.223
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| CAS # |
1365267-35-1
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| Related CAS # |
Rivaroxaban impurity 3
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| PubChem CID |
56943041
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| Appearance |
Solid powder
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| Hydrogen Bond Donor Count |
2
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
44
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| Complexity |
1010
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| Defined Atom Stereocenter Count |
2
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| SMILES |
C1COCC(=O)N1C2=CC=C(C=C2)N3C[C@@H](OC3=O)CNC(=O)NC[C@H]4CN(C(=O)O4)C5=CC=C(C=C5)N6CCOCC6=O
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| InChi Key |
RMYVVSKNFAMRGJ-ZEQRLZLVSA-N
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| InChi Code |
InChI=1S/C29H32N6O9/c36-25-17-41-11-9-32(25)19-1-5-21(6-2-19)34-15-23(43-28(34)39)13-30-27(38)31-14-24-16-35(29(40)44-24)22-7-3-20(4-8-22)33-10-12-42-18-26(33)37/h1-8,23-24H,9-18H2,(H2,30,31,38)/t23-,24-/m0/s1
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| Chemical Name |
1,3-bis[[(5S)-2-oxo-3-[4-(3-oxomorpholin-4-yl)phenyl]-1,3-oxazolidin-5-yl]methyl]urea
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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 | 1.6431 mL | 8.2156 mL | 16.4312 mL | |
| 5 mM | 0.3286 mL | 1.6431 mL | 3.2862 mL | |
| 10 mM | 0.1643 mL | 0.8216 mL | 1.6431 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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