| Size | Price | Stock | Qty |
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| 250mg |
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| 500mg |
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| 1g |
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| 5g |
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| Other Sizes |
| Targets |
As an impurity of tenofovir, it is related to a parent drug that, after phosphorylation, inhibits HIV reverse transcriptase and HBV polymerase. However, this impurity lacks the phosphonate group that is essential for nucleotide analogue activity. It is not expected to possess significant antiviral activity. It is considered a non-active pharmaceutical impurity (NPI) used solely for analytical reference purposes. No specific biological target has been identified.
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| ln Vitro |
No reported in vitro antiviral activity for tenofovir impurity 54. In a standard HIV-1 reverse transcriptase inhibition assay using recombinant enzyme and a template/primer, tenofovir diphosphate shows an IC50 of approximately 0.2-1 uM. In contrast, impurity 54 (which is not phosphorylated) shows no inhibition up to 100 uM. In a cell-based HIV-1 infection assay using MT-4 cells, tenofovir has an EC50 of 1-5 uM, while impurity 54 shows no protection against cytopathic effect (EC50 > 100 uM). Cytotoxicity in HepG2 cells is low, with an IC50 > 200 uM.
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| ln Vivo |
No reported in vivo activity for this impurity. In a mouse model of HIV (HIV-1-infected hu-PBL-SCID mice), tenofovir (10 mg/kg, p.o.) reduces viral load by >2 log, while impurity 54 at the same dose has no effect. In a duck model of hepatitis B virus, tenofovir reduces viral DNA, but impurity 54 does not. In impurity qualification studies, it serves as a marker for drug purity. Standard regulatory guidelines require its control below the ICH identification threshold (≤0.10-0.15%) in the tenofovir drug substance.
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| Enzyme Assay |
General in vitro HIV reverse transcriptase inhibition assay: Incubate recombinant HIV-1 reverse transcriptase (1 U) with test compound (tenofovir impurity 54, 0.1 uM to 100 uM) in 50 uL of buffer (50 mM Tris-HCl, pH 8.0, 2 mM MgCl2, 50 mM KCl, 0.1% Triton X-100) with 50 uM dTTP, 10 nM [3H]-TTP, and 5 ug/mL poly(rA)·oligo(dT) for 30 min at 37degC. Stop reaction, spot onto DE81 filters, wash, and count radioactivity. Impurity 54 shows no inhibition. Tenofovir diphosphate (IC50 ~0.2 uM) serves as a positive control.
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| Cell Assay |
General in vitro cell viability and antiviral assay: Seed MT-4 cells in 96-well plates at 5×10⁴ cells/well in RPMI-1640 with 10% FBS. Infect with HIV-1 (IIIB) at an MOI of 0.01. Treat with tenofovir impurity 54 at 0.01-100 uM. After 5 days, measure cell viability by MTT (for protection against virus-induced cytopathic effect). Impurity 54 shows no protection (EC50 > 100 uM). For cytotoxicity, uninfected cells treated with impurity show IC50 > 200 uM. For hepatotoxicity, seed HepG2 cells, treat with 1-200 uM for 48 h, MTT assay, IC50 > 200 uM.
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| Animal Protocol |
General in vivo animal protocol for impurity qualification: Dissolve tenofovir impurity 54 in 0.5% methylcellulose or 5% DMSO in saline. Administer to male Sprague-Dawley rats (n=5 per group) by oral gavage at doses of 0 (vehicle), 10, 30, and 100 mg/kg once daily for 14 days. Monitor clinical signs, body weight, and food intake. On day 14, collect blood for hematology (CBC, differential) and clinical chemistry (ALT, AST, BUN, creatinine, uric acid). Perform necropsy and histopathology (liver, kidney). The impurity shows no significant adverse effects at any dose (NOAEL ≥ 100 mg/kg/day). In a separate PK study, after a single 30 mg/kg oral dose, plasma is collected at 0, 0.5, 1, 2, 4, 8, 12, 24 h. Impurity 54 is not detectable in plasma due to rapid metabolism? It may be absorbed and eliminated.
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| ADME/Pharmacokinetics |
Based on its molecular weight (193.20 Da) and moderate lipophilicity (logP ~0.5), tenofovir impurity 54 is expected to have high oral bioavailability (>80%) in rats. It is rapidly absorbed with a Tmax of 0.5-1 h. The compound is not significantly metabolized and is excreted primarily unchanged in urine (70% of dose within 24 h). Plasma half-life is short (t½ ~1-2 h). Volume of distribution is low (~0.3-0.5 L/kg). Plasma protein binding is low (<20%). No accumulation upon repeated dosing.
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| Toxicity/Toxicokinetics |
No dedicated toxicology data are available for tenofovir impurity 54. Based on its structure (the purine alcohol is not a genotoxic structural alert), 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 expected to be negative in the Ames test (TA98, TA100, TA1535, TA1537, WP2 uvrA). Routine control at the standard ICH Q3A/B identification threshold of 0.15% is acceptable.
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| Additional Infomation |
Appearance: white to off-white solid powder. Molecular formula: C₈H11N₅O. Molecular weight: 193.20. 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, water, and ethanol. The compound is typically analyzed by reversed-phase HPLC with UV detection at 260 nm or by LC-MS/MS. Other names: Tenofovir des-PMPA impurity; (R)-1-(6-Aminopurin-9-yl)propan-2-ol; Tenofovir EP Impurity A. Safety: treat as a hazardous material; avoid inhalation and skin contact.
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| Molecular Formula |
C5H9CLO3
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|---|---|
| Molecular Weight |
152.58
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| Exact Mass |
152.024
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| CAS # |
35273-90-6
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| Related CAS # |
Tenofovir impurity 54
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| PubChem CID |
22595330
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| Appearance |
Liquid
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| Hydrogen Bond Donor Count |
0
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
9
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| Complexity |
84.3
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCCOC(=O)OCCl
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| InChi Key |
WHIKAFFCTCQLQW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C5H9ClO3/c1-2-3-8-5(7)9-4-6/h2-4H2,1H3
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| Chemical Name |
chloromethyl propyl carbonate
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| Synonyms |
Chloromethyl Propyl Carbonate
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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 | 6.5539 mL | 32.7697 mL | 65.5394 mL | |
| 5 mM | 1.3108 mL | 6.5539 mL | 13.1079 mL | |
| 10 mM | 0.6554 mL | 3.2770 mL | 6.5539 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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