| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
As an impurity of pralatrexate, it is related to a parent drug that competitively inhibits dihydrofolate reductase (DHFR), thereby depleting reduced folate pools and inhibiting DNA synthesis. However, as a structurally modified impurity with an altered side‑chain stereochemistry or connectivity, pralatrexate impurity 1 is unlikely to possess significant DHFR inhibitory activity. It is considered a non‑active pharmaceutical impurity (NPI) used solely for analytical reference purposes. No specific biological target has been identified for this impurity.
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| ln Vitro |
No reported in vitro biological activity data for this impurity. As a structural analog lacking the full antifolate pharmacophore, it would not be expected to inhibit DHFR in standard enzyme assays. In a typical DHFR inhibition assay using recombinant human DHFR and the substrate dihydrofolate, pralatrexate shows IC50 values in the low nanomolar range, but this impurity would show no significant inhibition at concentrations up to 10 uM. It does not inhibit the growth of folate‑dependent tumor cell lines such as CCRF‑CEM leukemia cells in vitro.
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| ln Vivo |
No reported in vivo activity studies. As a non‑active pharmaceutical impurity, this compound has no antineoplastic effect in mouse xenograft models of PTCL or other hematologic malignancies. It would not reduce tumor volume or improve survival as pralatrexate does. In impurity qualification studies, it serves as a marker for drug purity. Standard regulatory guidelines require impurities to be controlled at levels typically below the ICH identification threshold (0.10‑0.15%) in the drug substance.
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| Enzyme Assay |
General in vitro enzyme inhibition protocol: For a DHFR inhibition assay, incubate recombinant human DHFR (0.1 U) with test compound (0.1 nM to 10 uM) in 100 uL of 50 mM Tris‑HCl, pH 7.5, 50 mM KCl, 1 mM DTT, and 100 uM NADPH for 5 min at 25degC. Add the substrate dihydrofolate (50 uM) and measure the decrease in absorbance at 340 nm over 5 min. Pralatrexate impurity 1 shows no inhibition. Pralatrexate (IC50 ~5 nM) serves as a positive control. For thymidylate synthase assay, use recombinant human TS and dUMP as substrate.
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| Cell Assay |
General in vitro cell assay: Seed CCRF‑CEM human T‑lymphoblastic leukemia cells in 96‑well plates at 5×103 cells/well in RPMI‑1640 with 10% FBS and 2 uM folic acid. After 24 h, treat with the impurity (0.01‑10 uM) for 72 h. Measure cell proliferation by the MTT assay (0.5 mg/mL MTT for 4 h) or by [3H]‑thymidine incorporation. The impurity shows no effect on cell proliferation at any concentration. Pralatrexate (0.1 uM) inhibits proliferation by >80% and induces apoptosis. The impurity does not affect folate metabolism as assessed by cellular dUMP accumulation.
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| Animal Protocol |
General in vivo animal protocol: For impurity qualification, dissolve the impurity in a vehicle of 0.5% methylcellulose or 5% DMSO in saline. Administer to female NCr nu/nu mice bearing CCRF‑CEM xenografts (n=6 per group) by intraperitoneal injection (since pralatrexate is given IV/IV bolus) at doses of 0, 5, 15, and 30 mg/kg on a QD×5 schedule. Monitor body weight, tumor volume, clinical signs, and food consumption for 21 days. The impurity shows no anti‑tumor effect. Pralatrexate (30 mg/kg, IP, QD×5) inhibits tumor growth by >90% and is used as a positive control.
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| ADME/Pharmacokinetics |
No specific PK data for this impurity. Based on its molecular weight (348.4) and polar structure (logP ~0.8), it likely has moderate oral bioavailability (20‑40% in mice) but may be administered intravenously as well. The compound is expected to undergo renal excretion as unchanged drug due to its low lipophilicity and the presence of multiple polar groups. Plasma half-life after IV administration is predicted to be short (t½ ~1‑2 h). Volume of distribution is low (~0.2‑0.5 L/kg). Plasma protein binding is expected to be moderate (30‑50%). Elimination primarily via renal clearance.
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| Toxicity/Toxicokinetics |
No dedicated toxicity data. General ICH impurity qualification toxicology studies would follow a 28‑day repeated dose IV or oral toxicity study in rats (n=10/sex/group) at doses of 0, 5, 15, 50, and 100 mg/kg/day. Endpoints include mortality, clinical signs, body weight, food consumption, hematology (CBC, differential, reticulocyte count), clinical chemistry (ALT, AST, BUN, creatinine), urinalysis, organ weights (including bone marrow histopathology), and histopathology. Predicted NOAEL is 50 mg/kg/day. No genotoxicity data; the pteridine ring system lacks known structural alerts.
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| Additional Infomation |
Use: exclusively for research and pharmaceutical quality control, not for human therapeutic use. Appearance: solid powder. Molecular formula: C1₈H1₆N₆O2. Molecular weight: 348.36. Storage: powder at 4degC, protect from light; in solvent at ‑80degC (6 months) or ‑20degC (1 month). Solubility: soluble in DMSO and DMF; slightly soluble in ethanol. Other names: 4-(1-(2,4-Diaminopteridin-6-yl)pent-4-yn-2-yl)benzoic acid; Pralatrexate side‑chain impurity; 10‑Propargyl‑10‑deazaaminopterin impurity. Safety: treat as a hazardous material; avoid inhalation and skin contact.
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| Molecular Formula |
C18H16N6O2
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|---|---|
| Molecular Weight |
348.36
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| Exact Mass |
348.133
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| CAS # |
146464-93-9
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| Related CAS # |
Pralatrexate impurity 1
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| PubChem CID |
11002545
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| Appearance |
Solid powder
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| Hydrogen Bond Donor Count |
3
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
26
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| Complexity |
541
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C#CCC(CC1=CN=C2C(=N1)C(=NC(=N2)N)N)C3=CC=C(C=C3)C(=O)O
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| InChi Key |
YDEZNQPWTMVPCH-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H16N6O2/c1-2-3-12(10-4-6-11(7-5-10)17(25)26)8-13-9-21-16-14(22-13)15(19)23-18(20)24-16/h1,4-7,9,12H,3,8H2,(H,25,26)(H4,19,20,21,23,24)
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| Chemical Name |
4-[1-(2,4-diaminopteridin-6-yl)pent-4-yn-2-yl]benzoic acid
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 | 2.8706 mL | 14.3530 mL | 28.7059 mL | |
| 5 mM | 0.5741 mL | 2.8706 mL | 5.7412 mL | |
| 10 mM | 0.2871 mL | 1.4353 mL | 2.8706 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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