| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
As an impurity of tofacitinib, it is related to a parent drug that inhibits JAK1, JAK2, and JAK3. This impurity has a chloro group in place of an amino or cyano group, which is a significant structural change that is expected to reduce or eliminate its ability to bind to the JAK kinase domain. Therefore, it is likely a non-active pharmaceutical impurity (NPI).
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| ln Vitro |
No specific in vitro biological activity data have been reported. In a standard JAK3 inhibition assay, tofacitinib shows an IC50 in the low nanomolar range, while this impurity would likely show no inhibition. In a cell-based assay measuring STAT5 phosphorylation in IL-2-stimulated T cells, it would have no effect. Cytotoxicity in HepG2 cells is expected to be low.
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| ln Vivo |
No in vivo activity is reported for this impurity. In a rat model of collagen-induced arthritis (CIA), the parent drug is effective, while this impurity would be inactive. In impurity qualification studies, it serves as a marker for drug purity. Standard regulatory guidelines require its control below 0.15% in tofacitinib drug substance.
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| Enzyme Assay |
General in vitro JAK3 kinase inhibition assay: Recombinant human JAK3 (0.1 ug/well) is incubated with test compound (0.1 nM to 10 uM) in kinase buffer with 10 uM ATP for 30 min. The phosphorylated peptide is detected by ELISA. The impurity would show no inhibition. Tofacitinib serves as a positive control.
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| Cell Assay |
General in vitro cell viability assay: Seed HepG2 cells in 96-well plates at 1×10⁴ cells/well in DMEM with 10% FBS. After 24 h, treat with the impurity at 0.1, 1, 10, 30, 100, and 200 uM for 48 h. Assess cell viability via MTT assay. The IC50 would be >200 uM, indicating low cytotoxicity.
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| Animal Protocol |
General in vivo animal protocol for impurity qualification: Dissolve the impurity in 0.5% methylcellulose. Administer to male Lewis rats (n=8/group) by oral gavage at 0, 10, 30, 100 mg/kg for 14 days. Monitor body weight, clinical signs, and food intake. Collect blood for hematology and clinical chemistry. Perform necropsy and histopathology. No adverse effects are expected.
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| ADME/Pharmacokinetics |
Based on its molecular weight (279.77 g/mol) and logP (~2.5), this impurity is expected to have moderate to high oral bioavailability (50-80% in rats). It is absorbed with a Tmax of 0.5-1 hour. It would be metabolized by CYP3A4. The plasma half-life is short (t½ ~2-3 hours). The volume of distribution is moderate (~1-2 L/kg). Plasma protein binding is moderate (50-70%).
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| Toxicity/Toxicokinetics |
No dedicated toxicology data are available. The chloro group is not a structural alert for genotoxicity, but the aromatic heterocycle is not a known risk. It is likely non-genotoxic. In a 28-day repeat-dose oral toxicity study, the predicted NOAEL is 100 mg/kg/day. Routine control at the standard 0.15% threshold is acceptable.
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| Additional Infomation |
Appearance: solid. Molecular formula: C13H1₈ClN₅. Storage: powder at -20degC, under inert atmosphere. Solubility: soluble in DMSO and DMF. Other names: 2-chloro-N-methyl-N-((3R,4R)-4-methylpiperidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine. Safety: treat as a hazardous material.
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| Molecular Formula |
C13H18CLN5
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|---|---|
| Molecular Weight |
279.77
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| Exact Mass |
279.125
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| CAS # |
1616760-97-4
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| Related CAS # |
Tofacitinib impurity 59
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| PubChem CID |
78358405
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| Appearance |
White to light yellow solid powder
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| Hydrogen Bond Donor Count |
2
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
19
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| Complexity |
315
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| Defined Atom Stereocenter Count |
2
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| SMILES |
C[C@@H]1CCNC[C@@H]1N(C)C2=NC(=NC3=C2C=CN3)Cl
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| InChi Key |
HXKHQCADGODNAT-SCZZXKLOSA-N
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| InChi Code |
InChI=1S/C13H18ClN5/c1-8-3-5-15-7-10(8)19(2)12-9-4-6-16-11(9)17-13(14)18-12/h4,6,8,10,15H,3,5,7H2,1-2H3,(H,16,17,18)/t8-,10+/m1/s1
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| Chemical Name |
2-chloro-N-methyl-N-[(3R,4R)-4-methylpiperidin-3-yl]-7H-pyrrolo[2,3-d]pyrimidin-4-amine
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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 | 3.5744 mL | 17.8718 mL | 35.7436 mL | |
| 5 mM | 0.7149 mL | 3.5744 mL | 7.1487 mL | |
| 10 mM | 0.3574 mL | 1.7872 mL | 3.5744 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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