| Size | Price | Stock | Qty |
|---|---|---|---|
| 500mg |
|
||
| 5g |
|
||
| Other Sizes |
| ln Vitro |
8-azaguanine reduces cell viability in a manner dependent on time, dose, and type of cell; MOLT3 cells being the most susceptible to its harmful effects (24-hour IC50: 100 μM). When comparing MOLT3 cells to CEM cells under the same conditions, 8-azaguanine-treated MOLT3 cells had higher CD26 expression (MIF) (65.4 vs. 18.7) [1].
8-Azaguanine induced a dose- and time-dependent decrease in cell viability in T-acute lymphoblastic leukemia cell lines (CEM and MOLT3). The MOLT3 cell line was more sensitive to its cytotoxic effects, with a 24-hour IC₅₀ of approximately 10 µM, compared to the CEM cell line which had a 24-hour IC₅₀ of approximately 100 µM. Treatment with 8-azaguanine induced morphological changes characteristic of apoptosis, including cellular shrinking, membrane blebbing, and DNA fragmentation. Flow cytometry analysis using Annexin V/PI staining confirmed an increase in the percentage of cells in early and late apoptosis/necrosis upon treatment. Furthermore, 8-azaguanine treatment stimulated an increase in CD26 cell surface expression, measured as Mean Fluorescence Intensity (MFI). This increase was more pronounced in MOLT3 cells (MFI increased from 18.7 ± 1.7 to 65.4 ± 1.3 at 25 µM) compared to CEM cells. This effect was specific to 8-azaguanine, as treatment with hydrogen peroxide (an oxidative stress inducer) did not significantly alter CD26 expression. The enzymatic activity of DPPIV (soluble form) in the culture medium supernatant did not show significant changes after 8-azaguanine treatment.[1] |
|---|---|
| Enzyme Assay |
The enzymatic activity of Dipeptidyl Peptidase IV (DPPIV) in culture medium supernatant was measured using a fluorimetric assay. The enzyme catalyzes the cleavage of the fluorogenic substrate Gly-Pro-4-methyl-2-naphthylamide (Gly-Pro-4-Me-2-NA), releasing the fluorescent product 4-methyl-2-naphthylamine (4-Me-2-NA).
The assay was performed in a 50 mmol/L Tris-HCl buffer at pH 8.3. The fluorescence intensity was measured at an excitation wavelength of 340 nm and an emission wavelength of 425 nm. One unit of DPPIV activity was defined as the amount of enzyme that produces 1 µmol of 4-Me-2-NA per minute under the assay conditions.[1] |
| Cell Assay |
Cells (CEM and MOLT3 T-acute lymphoblastic leukemia cell lines) were cultured in RPMI-1640 medium supplemented with fetal calf serum.
For proliferation and viability assays, cells were incubated in the absence or presence of increasing concentrations of 8-azaguanine (ranging from 10 to 200 µM) for 24, 48, 72, and 96 hours. Cell viability was assessed at each time point by trypan blue exclusion and cell counting using a hemocytometer. Cell morphology was evaluated by light microscopy examination of May-Grünwald-Giemsa stained slides. Apoptosis and necrosis were analyzed by flow cytometry using Annexin V-FITC and propidium iodide (PI) staining. Cells were washed, incubated with Annexin V-FITC and PI in a binding buffer for 10 minutes at 4°C, then washed again and analyzed. For analysis of CD26 cell surface expression, cells were incubated with a PE-conjugated anti-CD26 monoclonal antibody for 10 minutes at room temperature, washed, and analyzed by flow cytometry to determine the percentage of positive cells and the Mean Fluorescence Intensity (MFI). Isotype-matched antibodies were used as negative controls.[1] |
| References | |
| Additional Infomation |
8-Zazaguanine is a triazolopyrimidine compound composed of 3,6-dihydro-7H-[1,2,3]triazolo[4,5-d]pyrimidine, with amino and oxygen substituents at positions 5 and 7, respectively. It possesses antimetabolite, antitumor, and EC 2.4.2.1 (purine nucleoside phosphorylase) inhibitory activities. It is a triazolopyrimidine compound and a nucleobase analog. 8-Zazaguanine is one of the early discovered purine analogs with antitumor activity. It has antimetabolite activity and is readily incorporated into ribonucleic acid. 8-Zazaguanine is a purine analog with potential antitumor activity. 8-Zazaguanine interferes with tRNA modification by competing with guanine for tRNA modification catalyzed by tRNA-guanine ribosyltransferase (tRNA-guanine transglycosylase). Alterations in tRNA-guanine modification are associated with cell differentiation and tumor transformation. 8-azaguanine can also inhibit the formation of the 43S and 80S initiation complex, thereby interfering with translation initiation and inhibiting protein synthesis. This drug can inhibit tumor cell growth and stimulate cell differentiation. (NCI04)
It is one of the early discovered purine analogs with antitumor activity. It is an antimetabolite that is readily incorporated into ribonucleic acid. 8-azaguanine is a purine analog with antitumor activity. It acts as an antimetabolite and is readily incorporated into ribonucleic acid (RNA), interfering with normal biosynthetic pathways and inhibiting cell growth. This study suggests that the drug-induced increase in CD26 expression on leukemia cells may be related to the sensitivity of cells to 8-azaguanine, suggesting that CD26 may play a role in the chemotherapy response to T-cell acute lymphoblastic leukemia. [1] |
| Molecular Formula |
C4H4N6O
|
|---|---|
| Molecular Weight |
152.1142
|
| Exact Mass |
152.044
|
| CAS # |
134-58-7
|
| PubChem CID |
135403646
|
| Appearance |
White to yellow solid powder
|
| Density |
2.6±0.1 g/cm3
|
| Melting Point |
>300 °C(lit.)
|
| Flash Point |
129.1ºC
|
| Vapour Pressure |
0.00149mmHg at 25°C
|
| Index of Refraction |
2.308
|
| LogP |
-0.71
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
0
|
| Heavy Atom Count |
11
|
| Complexity |
225
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
LPXQRXLUHJKZIE-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C4H4N6O/c5-4-6-2-1(3(11)7-4)8-10-9-2/h(H4,5,6,7,8,9,10,11)
|
| Chemical Name |
5-amino-2,6-dihydrotriazolo[4,5-d]pyrimidin-7-one
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
DMSO : ~2 mg/mL (~13.15 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 0.24 mg/mL (1.58 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 2.4 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 0.24 mg/mL (1.58 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 2.4 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 0.24 mg/mL (1.58 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 6.5742 mL | 32.8709 mL | 65.7419 mL | |
| 5 mM | 1.3148 mL | 6.5742 mL | 13.1484 mL | |
| 10 mM | 0.6574 mL | 3.2871 mL | 6.5742 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
