| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| Targets |
DNA polymerases; HIV-1 reverse transcriptase[1]
HIV-1 reverse transcriptase (RT) and HBV DNA polymerase. It is a substrate for HIV-1 RT, where it competes with the natural substrate deoxyadenosine 5'-triphosphate (dATP). Once incorporated into viral DNA, it terminates chain elongation. |
|---|---|
| ln Vitro |
Tenofovir diphosphate functions as a DNA pol inhibitor. Tenofovir-diphosphate exhibits mild inhibitory effects on DNA polymerases (pol) α, δ, and ʫ. Its Ki for PMPApp (PMPAppKi) is 10.2, 10.2, and 15.6, respectively, in relation to the Km for dATP (dATPKm)[1].
In vitro, Tenofovir diphosphate is a potent inhibitor of HIV-1 reverse transcriptase. It inhibits the DNA-dependent DNA polymerase activity with a Ki of 1.55 uM and the RNA-dependent DNA polymerase activity with a Ki of 0.022 uM. This more potent inhibition of the RNA-dependent activity is key to its anti-HIV-1 mechanism. It also inhibits HBV polymerase. |
| ln Vivo |
The in vivo activity is achieved after administration of the parent prodrugs (TDF or TAF). These prodrugs are absorbed and converted to tenofovir, which is then phosphorylated intracellularly to the active tenofovir diphosphate. The active form has a long intracellular half-life (over 60 hours in peripheral blood mononuclear cells), allowing for once-daily dosing. It is highly effective at suppressing viral replication in HIV and HBV patients.
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| Enzyme Assay |
Tenofovir diphosphate (PMPApp) is a weak inhibitor of DNA polymerases (pol) α, δ, and ɛ*, with values for the Ki for PMPApp (PMPAppKi) relative to the Km for dATP (dATPKm) of 10.2, 10.2, and 15.2, respectively. Its incorporation into DNA was about 1,000-fold less efficient than that of dATP, with PMPAppKm values 350-, 2,155-, and 187-fold higher than dATPKm values for pol α, δ, and ɛ*, respectively[1].
A standard biochemical assay is a reverse transcriptase inhibition assay. In this cell-free system, purified HIV-1 RT is incubated with an RNA template, a complementary DNA primer, dNTPs (including a radiolabeled one), and varying concentrations of Tenofovir diphosphate. The enzyme incorporates the nucleotides, extending the DNA primer. The reaction is stopped and the radiolabeled, extended DNA product is captured and measured. The IC₅0 or Ki is then calculated. |
| Cell Assay |
The active anabolite itself is not applied directly to cells. The cellular activity is assessed using the parent prodrugs. A standard protocol involves infecting a human T-cell line (e.g., MT-2 or CEM) with HIV-1. The cells are then treated with serial dilutions of tenofovir (or its prodrugs). After 5-7 days of culture, antiviral efficacy is measured by quantifying a viral protein like p24 in the supernatant via ELISA. Alternatively, the inhibition of viral cytopathic effect is measured using a cell viability dye like XTT.
|
| Animal Protocol |
In vivo studies are performed with the parent prodrugs, TDF or TAF. A typical animal study for HIV research uses a humanized mouse model (e.g., NSG mice engrafted with human CD34+ cells) infected with HIV-1. The compound is administered daily via oral gavage. The primary endpoint is the viral load in the plasma, measured by RT-qPCR. For HBV research, a mouse model with persistent HBV replication (e.g., AAV-HBV mouse model) is used.
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| ADME/Pharmacokinetics |
PK data applies to the parent prodrugs. Tenofovir disoproxil fumarate (TDF) is a prodrug that is rapidly converted to tenofovir. Tenofovir alafenamide (TAF) is a newer prodrug designed for increased stability and more efficient delivery to lymphocytes. TAF achieves higher intracellular concentrations of tenofovir diphosphate at a lower dose, reducing systemic exposure. Tenofovir is cleared by the kidneys via glomerular filtration and active tubular secretion.
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| Toxicity/Toxicokinetics |
The active anabolite itself is not used in vivo, so its direct toxicity is not profiled. The safety of tenofovir therapy is well-established. TDF has been associated with renal toxicity (e.g., Fanconi syndrome) and decreases in bone mineral density in some patients. The newer prodrug TAF has a more favorable renal and bone safety profile due to its targeted delivery. Both are generally well-tolerated.
|
| References | |
| Additional Infomation |
Tenofovir is a nucleotide analog, which is different from a nucleoside analog (NRTI) as it already has a monophosphate group attached, requiring only two additional phosphorylation steps to become active. The disodium salt form is provided as a research standard. The triphosphate form is key for mechanism of action studies.
|
| Molecular Formula |
C9H16N5NA2O10P3
|
|---|---|
| Molecular Weight |
493.151665687561
|
| Exact Mass |
492.99
|
| CAS # |
2738719-07-6
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| Related CAS # |
Tenofovir diphosphate;166403-66-3;Tenofovir diphosphate triethylamine;2122333-63-3
|
| PubChem CID |
163335230
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| Appearance |
White to off-white solid powder
|
| Hydrogen Bond Donor Count |
5
|
| Hydrogen Bond Acceptor Count |
14
|
| Rotatable Bond Count |
9
|
| Heavy Atom Count |
29
|
| Complexity |
662
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
C[C@H](CN1C=NC2=C(N=CN=C21)N)OCP(=O)(O)OP(=O)(O)OP(=O)(O)O.[Na].[Na]
|
| InChi Key |
PUFMXPADRYEPCZ-QYCVXMPOSA-N
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| InChi Code |
InChI=1S/C9H16N5O10P3.2Na/c1-6(2-14-4-13-7-8(10)11-3-12-9(7)14)22-5-25(15,16)23-27(20,21)24-26(17,18)19;;/h3-4,6H,2,5H2,1H3,(H,15,16)(H,20,21)(H2,10,11,12)(H2,17,18,19);;/t6-;;/m1../s1
|
| Chemical Name |
C[C@H](CN1C=NC2=C(N=CN=C21)N)OCP(=O)(O)OP(=O)(O)OP(=O)(O)O.[Na].[Na]
|
| 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, 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)
|
| Solubility (In Vitro) |
H2O : 100 mg/mL (203.61 mM)
|
|---|---|
| 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 | 2.0278 mL | 10.1389 mL | 20.2778 mL | |
| 5 mM | 0.4056 mL | 2.0278 mL | 4.0556 mL | |
| 10 mM | 0.2028 mL | 1.0139 mL | 2.0278 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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