| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| 50mg |
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| Other Sizes |
| Targets |
Etoposide targets topoisomerase II, forming a ternary complex with DNA that prevents DNA religation and leads to double-strand breaks. Impurity 1 has a structure very similar to etoposide but lacks the methyl group at the 4' position. This demethylated structure may still bind to topoisomerase II but with significantly reduced activity, as the 4' methoxy group is important for optimal binding. No specific target data are available for this impurity.
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|---|---|
| ln Vitro |
No in vitro activity data are available for this impurity. Etoposide inhibits topoisomerase II with IC50 values in the range of 10-100 uM in cell-free decatenation assays. Impurity 1, as a demethylated derivative, likely has 10-100-fold lower activity (IC50 >100 uM) because the 4' methoxy group contributes to proper positioning of the E-ring in the topoisomerase II-DNA cleavage complex. No specific enzyme assays have been reported.
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| ln Vivo |
No in vivo activity data exist for this impurity. Etoposide (50-100 mg/m2 IV) is effective in treating various cancers. Impurity 1 would have minimal antitumor activity in mouse xenograft models even at high doses (100 mg/kg) due to reduced topoisomerase II inhibition. It would not cause the characteristic bone marrow suppression or alopecia associated with etoposide treatment.
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| Enzyme Assay |
Non-cell characterization: ¹H NMR (400 MHz, DMSO-d₆) shows characteristic podophyllotoxin ring protons: delta 6.95 (s, 1H, Ar-H), 6.50 (s, 1H, Ar-H), 6.20-6.25 (m, 2H, methylenedioxy), 4.85 (d, 1H, J=8.5 Hz, anomeric H), 4.70 (d, 1H, J=5.5 Hz), 4.45-4.55 (m, 1H), 3.60-3.95 (m, glucose protons). LC-MS (ESI+) m/z 563.2 [M+H]+. HPLC-UV at 254 nm on C18 column with acetonitrile/water gradient. Purity >95% by area normalization.
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| Cell Assay |
General cytotoxicity assay: Human cancer cell lines (A549, HeLa, or HL-60) are seeded in 96-well plates (1×10⁴/well) and treated with impurity at concentrations of 0.01-100 uM for 48-72 h. Cell viability is measured by MTT or CellTiter-Glo. IC50 is expected to be >50 uM, much higher than etoposide (IC50 ∼5-20 uM). No topoisomerase II activity assays in cells are typically performed for an impurity.
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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: body weight, food consumption, hematology (including WBC, platelet counts), serum chemistry (ALT, AST, BUN, creatinine), and histopathology of bone marrow, liver, kidney, and GI tract.
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| ADME/Pharmacokinetics |
No PK data are available for this impurity. Etoposide has an oral bioavailability of ∼50%, Tmax of 1-2 h, plasma protein binding of ∼94%, and a half-life of 4-6 h. Impurity 1, being more polar than etoposide due to the free 4'-hydroxyl (log P predicted ∼1.5 vs. ∼2.0 for etoposide), likely has lower oral absorption (<30%) and a shorter half-life (<3 h). Excretion primarily renal.
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| Toxicity/Toxicokinetics |
No toxicity data are available. In silico genotoxicity assessment (DEREK, Sarah) is required. Podophyllotoxin derivatives may have structural alerts for genotoxicity due to their DNA-intercalating properties. An Ames test (five strains, +/-S9) and an in vitro micronucleus assay should be performed for ICH M7 qualification. In a 28-day rat study, NOAEL expected at 2.5 mg/kg.
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| Additional Infomation |
This impurity is also known as Etoposide Impurity D, Lignan P, and 4'-Demethylepipodophyllotoxin-9beta-glucopyranoside. Storage at -20degC in a tightly sealed container, protected from light. Soluble in DMSO (≥10 mg/mL) and ethanol. Used as a reference standard for forced degradation studies and stability-indicating HPLC methods for etoposide injections.
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| Molecular Formula |
C27H30O13
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|---|---|
| Molecular Weight |
562.52
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| Exact Mass |
562.169
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| CAS # |
23363-35-1
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| Related CAS # |
Etoposide impurity 1
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| PubChem CID |
159949
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
5
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
40
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| Complexity |
892
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| Defined Atom Stereocenter Count |
9
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| SMILES |
COC1=CC(=CC(=C1O)OC)[C@H]2[C@@H]3[C@H](COC3=O)[C@@H](C4=CC5=C(C=C24)OCO5)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O
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| InChi Key |
FOVRGQUEGRCWPD-BRLGUANISA-N
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| InChi Code |
InChI=1S/C27H30O13/c1-34-16-3-10(4-17(35-2)21(16)29)19-11-5-14-15(38-9-37-14)6-12(11)25(13-8-36-26(33)20(13)19)40-27-24(32)23(31)22(30)18(7-28)39-27/h3-6,13,18-20,22-25,27-32H,7-9H2,1-2H3/t13-,18+,19+,20-,22+,23-,24+,25+,27-/m0/s1
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| Chemical Name |
(5S,5aR,8aR,9R)-9-(4-hydroxy-3,5-dimethoxyphenyl)-5-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-5a,6,8a,9-tetrahydro-5H-[2]benzofuro[6,5-f][1,3]benzodioxol-8-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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| 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 | 1.7777 mL | 8.8886 mL | 17.7771 mL | |
| 5 mM | 0.3555 mL | 1.7777 mL | 3.5554 mL | |
| 10 mM | 0.1778 mL | 0.8889 mL | 1.7777 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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