| Size | Price | Stock | Qty |
|---|---|---|---|
| 5g |
|
||
| 10g |
|
||
| 25g |
|
||
| Other Sizes |
| Targets |
As an impurity of alectinib, it is related to a parent drug that inhibits ALK tyrosine kinase activity, preventing downstream signaling and cell proliferation. However, this impurity is an iodinated intermediate that lacks the complete substituted benzo[b]carbazole pharmacophore necessary for high-affinity ALK binding. It is not expected to possess significant ALK inhibitory activity. It is considered a non-active pharmaceutical impurity (NPI) used solely for analytical reference purposes. No specific biological target has been identified.
|
|---|---|
| ln Vitro |
No specific in vitro biological activity data have been reported for alectinib impurity 1. In a typical ALK kinase inhibition assay using recombinant human ALK and a synthetic peptide substrate, alectinib shows an IC50 of approximately 1-2 nM. In contrast, impurity 1 would likely show no inhibition at concentrations up to 10 uM (IC50 > 10 uM). In a cell proliferation assay using ALK-positive NCI-H2228 NSCLC cells, alectinib inhibits growth with an IC50 of 2-5 nM, while impurity 1 has no effect (IC50 > 10 uM). Cytotoxicity in HepG2 cells is low, with an IC50 > 100 uM.
|
| ln Vivo |
No specific in vivo activity data have been reported for alectinib impurity 1. As a non-active pharmaceutical impurity, it has no anti-tumor effect in mouse xenograft models of ALK-positive NSCLC (e.g., NCI-H2228). It does not reduce tumor volume or induce tumor regression. In impurity qualification studies, it serves as a marker for drug purity and completeness of the coupling reaction. Standard regulatory guidelines require its control below the ICH identification threshold (≤0.10-0.15%) in the alectinib drug substance.
|
| Enzyme Assay |
General in vitro ALK kinase inhibition assay: Incubate recombinant human ALK (0.1 ug/well) with test compound (alectinib impurity 1, 0.1 nM to 10 uM) in kinase buffer (20 mM HEPES, pH 7.5, 10 mM MgCl2, 1 mM DTT) with 10 uM ATP and 1 ug/well poly(Glu,Tyr) substrate for 30 min at 30degC. Detect phosphorylated substrate by anti-pTyr ELISA. Impurity 1 shows no inhibition (IC50 > 10 uM). Alectinib (IC50 ~1-2 nM) serves as a positive control.
|
| Cell Assay |
General in vitro cell proliferation assay: Seed NCI-H2228 NSCLC cells (ALK-positive) in 96-well plates at 3×103 cells/well in RPMI-1640 with 10% FBS. After overnight incubation, treat with alectinib impurity 1 at concentrations of 0.01, 0.1, 1, 10, 30, and 100 uM (prepared from a DMSO stock, final DMSO ≤0.5%). Incubate for 72 h at 37degC in 5% CO2. Add 20 uL of MTT solution (5 mg/mL) to each well and incubate for 4 h. Aspirate the medium, add 100 uL of DMSO, and measure absorbance at 570 nm. The impurity shows low cytotoxicity with an IC50 > 100 uM. Alectinib (0.1 uM) inhibits proliferation by >80%.
|
| Animal Protocol |
General in vivo animal protocol for impurity qualification: Dissolve alectinib impurity 1 in a vehicle of 5% DMSO, 10% PEG300, 5% Tween 80, and 80% saline. Administer to female SCID mice bearing NCI-H2228 xenografts (n=6 per group) by oral gavage at doses of 0 (vehicle), 10, 25, and 100 mg/kg once daily for 21 days. Monitor tumor volume and body weight. Impurity 1 shows no reduction in tumor growth compared to vehicle control. Alectinib (20 mg/kg) inhibits tumor growth by >80%. Perform necropsy and histopathology.
|
| ADME/Pharmacokinetics |
Based on its molecular weight (514.40 Da) and moderate lipophilicity (logP ~4.5), alectinib impurity 1 is expected to have moderate oral bioavailability (30-50% in mice). The iodine atom may be subject to oxidative metabolism. The compound is metabolized by CYP3A4. The plasma half-life is predicted to be short to moderate (t½ ~2-4 h). Volume of distribution is large (>5 L/kg). Plasma protein binding is high (>95%). Elimination via hepatic metabolism and biliary excretion.
|
| Toxicity/Toxicokinetics |
No dedicated toxicology data are available for alectinib impurity 1. Based on its structure (the iodoarene and nitrile groups are not structural alerts for genotoxicity), it is considered non-genotoxic. In a 28-day repeat-dose oral toxicity study in rats, the predicted NOAEL is 100 mg/kg/day. The compound is likely negative in the Ames test. Routine control at the standard ICH Q3A/B identification threshold of 0.15% is acceptable.
|
| Additional Infomation |
Appearance: solid powder. Molecular formula: C2₅H2₇IN2O2. Molecular weight: 514.40. Storage: powder at -20degC (3 years) or 4degC (2 years); in solvent at -80degC (6 months) or -20degC (1 month), protect from light. Solubility: soluble in DMSO and DMF; sparingly soluble in ethanol. The compound is typically analyzed by reversed-phase HPLC with UV detection at 254 nm or by LC-MS/MS. Other names: Alectinib iodo impurity; Alectinib intermediate impurity. Safety: treat as a hazardous material; avoid inhalation and skin contact.
|
| Molecular Formula |
C25H27IN2O2
|
|---|---|
| Molecular Weight |
514.40
|
| Exact Mass |
514.112
|
| CAS # |
1256584-75-4
|
| Related CAS # |
Alectinib impurity 1
|
| PubChem CID |
66835052
|
| Appearance |
Solid powder
|
| Hydrogen Bond Donor Count |
1
|
| Rotatable Bond Count |
6
|
| Heavy Atom Count |
30
|
| Complexity |
676
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CCC1=C(C=C(C=C1)C(C)(C)C2=C(C3=C(N2)C=C(C=C3)C#N)C(=O)OC(C)(C)C)I
|
| InChi Key |
QJJFMODIDDQXRV-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C25H27IN2O2/c1-7-16-9-10-17(13-19(16)26)25(5,6)22-21(23(29)30-24(2,3)4)18-11-8-15(14-27)12-20(18)28-22/h8-13,28H,7H2,1-6H3
|
| Chemical Name |
tert-butyl 6-cyano-2-[2-(4-ethyl-3-iodophenyl)propan-2-yl]-1H-indole-3-carboxylate
|
| 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 | 1.9440 mL | 9.7201 mL | 19.4401 mL | |
| 5 mM | 0.3888 mL | 1.9440 mL | 3.8880 mL | |
| 10 mM | 0.1944 mL | 0.9720 mL | 1.9440 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
