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| Targets |
The primary target of Az-Dcme is the HIV-1 reverse transcriptase (RT), an RNA-dependent DNA polymerase that is essential for the replication of HIV. HIV-1 RT converts the viral single-stranded RNA genome into double-stranded DNA, which is then integrated into the host cell's genome. As a nucleoside analog, Az-Dcme is a prodrug that requires intracellular activation. It is taken up by cells and phosphorylated by cellular kinases to its active triphosphate form, Az-Dcme-TP. The active triphosphate then acts as a competitive inhibitor of the natural substrate, deoxycytidine triphosphate (dCTP), for binding to the HIV-1 RT. When Az-Dcme-TP is incorporated into the growing viral DNA chain by the RT, it lacks a 3'-hydroxyl group, which is required for the formation of the phosphodiester bond with the next nucleotide. This causes chain termination, preventing the completion of the viral DNA synthesis and thus inhibiting viral replication. The selectivity of Az-Dcme for HIV-1 RT over human DNA polymerases is a key feature of its activity. Its potency is demonstrated by its low EC50 values against HIV-1 in cell culture. This makes it a valuable tool for studying the mechanism of NRTIs and the development of resistance.
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| ln Vitro |
Additionally efficient against HIV-2 in lymphocytes is AzddMeC (CS-92). There was no discernible impact on the replication of the Friend murine virus, HSV types 1 and 2, or coxsackievirus B4. HIV-1 reverse transcriptase and AzddMeC 5'-triphosphate exhibit competitive inhibition in their interaction (inhibition constant Kis is 9.3 nM) [1].
In vitro studies have characterized Az-Dcme as a potent and selective inhibitor of HIV-1 replication. Its activity is typically measured using HIV-1-infected cell cultures. In a standard assay, human peripheral blood mononuclear (PBM) cells or macrophages are infected with HIV-1 and then treated with various concentrations of Az-Dcme. After a period of incubation (e.g., 5-7 days), viral replication is measured by quantifying the levels of HIV-1 p24 antigen in the culture supernatant or by measuring the activity of the viral reverse transcriptase. The EC50, which is the concentration that inhibits 50% of viral replication, is determined from the dose-response curve. Az-Dcme has been shown to inhibit HIV-1 replication in human PBM cells with an EC50 of 9 nM and in human macrophages with an EC50 of 6 nM. These values demonstrate its high potency. In addition to its antiviral activity, its selectivity is confirmed by testing its cytotoxicity in uninfected cells. The CC50, which is the concentration that causes 50% cytotoxicity, is typically much higher than the EC50, indicating a favorable selectivity index. These in vitro studies are crucial for characterizing the antiviral activity and safety of Az-Dcme. |
| ln Vivo |
Male rhesus monkeys were given 60 mg/kg of the compound intravenously and orally, after which the pharmacokinetics of AzddMeC were described. In monkeys, the main metabolite of AzddMeC is 3'-azido-3'-deoxythymidine (AZT). Serum concentrations of AzddMeC have a terminal half-life of 0.5 to 1.3 hours and decrease quickly in a biexponential manner. The main routes for AzddMeC clearance are renal excretion of unmodified nucleosides and metabolic deamination to produce AZT. It has a 26% oral bioavailability[2].
In vivo activity of Az-Dcme has been demonstrated in animal models of HIV infection, such as the SCID-hu mouse model, where human fetal thymus and liver tissues are implanted into immunodeficient mice and then infected with HIV. In such models, administration of Az-Dcme has been shown to reduce viral load. Its oral bioavailability is a key feature that makes it a promising candidate for development. In a typical study, the compound would be administered to the animals orally, and the viral load in the blood or tissues would be measured over time. The reduction in viral load would be compared to a vehicle control group. These in vivo studies are essential for validating the antiviral activity of Az-Dcme and for providing proof-of-concept for its potential as a therapeutic agent. However, Az-Dcme is a research compound, and its development may have been discontinued in favor of other NRTIs. Its primary use is as a tool for studying HIV-1 reverse transcriptase and antiviral drug development. |
| Enzyme Assay |
The in vitro enzyme assay for Az-Dcme is a standard HIV-1 reverse transcriptase (RT) inhibition assay. In this assay, recombinant HIV-1 RT is incubated with a template-primer (e.g., a synthetic RNA template and a DNA primer), the four deoxynucleoside triphosphates (dNTPs), one of which is radiolabeled, and varying concentrations of the active triphosphate of Az-Dcme (Az-Dcme-TP). The RT catalyzes the incorporation of the dNTPs into the growing DNA chain, and the incorporation of the radiolabeled nucleotide is measured. The inhibition of RT activity by Az-Dcme-TP is determined by measuring the reduction in the incorporation of the radiolabeled nucleotide. The IC50, which is the concentration that inhibits 50% of RT activity, is determined from the dose-response curve. This assay is a direct measure of the compound's potency at its molecular target. To confirm the mechanism of action, the ability of Az-Dcme-TP to act as a chain terminator can be demonstrated by sequencing the DNA product. These enzyme assays are crucial for characterizing the mechanism of action of NRTIs.
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| Cell Assay |
In vitro cell-based assays for Az-Dcme are used to measure its antiviral activity against HIV-1. The most common assay is the HIV-1 replication assay in human PBM cells or macrophages. In this assay, cells are isolated from human blood and stimulated with phytohemagglutinin (PHA) and interleukin-2 (IL-2). The cells are then infected with HIV-1 (e.g., the laboratory strain IIIB) in the presence of varying concentrations of Az-Dcme. After a period of incubation (typically 5-7 days), the culture supernatant is collected, and the amount of HIV-1 p24 antigen is measured by ELISA. The p24 antigen is a viral core protein, and its level is a direct measure of viral replication. The EC50 is determined from the dose-response curve. For Az-Dcme, the EC50 is 9 nM in PBM cells and 6 nM in macrophages. To measure cytotoxicity, the same cells are cultured without virus and treated with Az-Dcme, and cell viability is measured using a dye such as MTT. The CC50 is determined, and the selectivity index (SI = CC50/EC50) is calculated. These cell-based assays are the standard for evaluating the antiviral activity of NRTIs.
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| Animal Protocol |
In vivo animal experiments for Az-Dcme are conducted to evaluate its antiviral efficacy and pharmacokinetic properties. A commonly used model is the SCID-hu mouse model. In this model, severe combined immunodeficient (SCID) mice are implanted with human fetal thymus and liver tissues (Thy/Liv) to create a humanized immune system. The mice are then infected with HIV-1, which replicates in the implanted human tissues. Az-Dcme is administered orally or by injection, and the viral load in the implanted tissues is measured by quantitative PCR or by measuring p24 antigen levels. The efficacy is determined by comparing the viral load in treated animals to that in vehicle-treated control animals. These studies provide evidence for the in vivo efficacy of Az-Dcme. The compound's oral bioavailability makes it suitable for oral administration in these studies. However, Az-Dcme is a research compound, and its development as a clinical candidate may have been discontinued. It is used as a tool for studying NRTI pharmacology and resistance.
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| ADME/Pharmacokinetics |
Az-Dcme is orally bioactive, which is a key feature for its potential as a therapeutic agent. Its pharmacokinetic (PK) properties have been studied in preclinical species. After oral administration, it is absorbed and distributed throughout the body. It is taken up by cells and phosphorylated to its active triphosphate. The active triphosphate has a long intracellular half-life, which contributes to its antiviral activity. Az-Dcme has a molecular weight of 266.26 g/mol and a molecular formula of C10H14N6O3. It is a nucleoside analog, and its chemical properties influence its PK. For research use, it is typically supplied as a solid and is soluble in DMSO. Its stability is ensured by storing it under recommended conditions (typically -20°C). A comprehensive PK profile is essential for understanding its in vivo efficacy and for designing dosing regimens. The compound's oral bioavailability makes it a convenient candidate for therapeutic use.
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| Toxicity/Toxicokinetics |
Detailed toxicity data for Az-Dcme is not provided in standard product descriptions, as it is a research compound. However, as a nucleoside analog, its potential toxicity would be related to its effects on mitochondrial DNA synthesis, which is a class effect of some NRTIs. The selectivity of Az-Dcme for HIV-1 RT over human DNA polymerases is a key factor in its safety profile. In preclinical studies, its safety would be assessed in rodents and dogs to determine its maximum tolerated dose and to identify any target organs of toxicity. Common side effects of NRTIs include gastrointestinal disturbances, headache, and, in some cases, mitochondrial toxicity, which can lead to lactic acidosis and hepatic steatosis. However, the specific toxicity profile of Az-Dcme has not been extensively characterized, as it is a research compound. For research use, it should be handled with standard laboratory safety precautions, using appropriate personal protective equipment (PPE). It is not intended for human use.
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| References | |
| Additional Infomation |
Az-Dcme is a research compound and is not approved for any clinical use. It is a potent and selective inhibitor of HIV-1 reverse transcriptase and HIV-1 replication, and it is orally bioactive. It has been studied as a potential treatment for HIV infection, but its development may have been discontinued in favor of other NRTIs that have better pharmacokinetic properties or safety profiles. As a nucleoside reverse transcriptase inhibitor (NRTI), its mechanism of action involves incorporation into the viral DNA and chain termination, which is a common mechanism for this class of drugs. Az-Dcme is used as a research tool to study the mechanism of HIV-1 reverse transcriptase, to investigate the development of drug resistance, and to screen for new NRTIs. Its high potency (EC50 of 9 nM in PBM cells and 6 nM in macrophages) makes it a valuable compound for these studies. It is also a useful tool for studying the pharmacology of nucleoside analogs. It is available for research purposes only and is not intended for human use.
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| Molecular Formula |
C10H14N6O3
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| Molecular Weight |
266.261
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| Exact Mass |
266.112
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| CAS # |
87190-79-2
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| PubChem CID |
64986
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| Appearance |
White to off-white solid powder
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| LogP |
-0.47
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
19
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| Complexity |
496
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| Defined Atom Stereocenter Count |
3
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| SMILES |
CC1=CN(C(=O)N=C1N)[C@H]2C[C@@H]([C@H](O2)CO)N=[N+]=[N-]
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| InChi Key |
GZSDAHQGNUAEBC-XLPZGREQSA-N
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| InChi Code |
InChI=1S/C10H14N6O3/c1-5-3-16(10(18)13-9(5)11)8-2-6(14-15-12)7(4-17)19-8/h3,6-8,17H,2,4H2,1H3,(H2,11,13,18)/t6-,7+,8+/m0/s1
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| Chemical Name |
4-amino-1-[(2R,4S,5S)-4-azido-5-(hydroxymethyl)oxolan-2-yl]-5-methylpyrimidin-2-one
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| Synonyms |
Az-Dcme AzddMeC CS-92
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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, 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) |
DMSO : ~200 mg/mL (~751.15 mM)
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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 | 3.7557 mL | 18.7786 mL | 37.5573 mL | |
| 5 mM | 0.7511 mL | 3.7557 mL | 7.5115 mL | |
| 10 mM | 0.3756 mL | 1.8779 mL | 3.7557 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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