Size | Price | Stock | Qty |
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50mg |
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100mg |
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250mg |
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500mg |
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1g |
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Purity: ≥98%
Puromycin aminonucleoside (formerly known as NSC 3056; PAN, NSC-3056), the aminonucleoside portion of the antibiotic puromycin, is commonly used in nephrosis animal models as well as in the study of human glomerular disease, and glomerular function and morphology. In vitro glomerular endothelial cells, puromycin aminonucleoside is used to investigate the relationship between cell permeability and endothelial glycosaminoglycan synthesis. Using Nox4 induction, PAN treatment dramatically elevated the oxidative stress level of podocytes. The oxidative stress-dependent modulation of ZO-1 by puromycin aminonucleoside leads to an increase in podocyte permeability.
Targets |
DPP-2; cytosol alanyl aminopeptidase
- Puromycin Aminonucleoside (NSC-3056) primarily targets podocytes and mesangial cells in the kidney, inducing apoptosis through mechanisms involving p53 activation and Bcl-2 family protein dysregulation [2] - It also interacts with organic cation transporter PMAT expressed in podocytes, contributing to nephrotoxicity [5] |
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ln Vitro |
Puromycin aminonucleoside (PAN) causes MC apoptosis, which is accompanied by a decrease in cell viability and an increase in the inflammatory response. ERRα overexpression exacerbates the apoptosis caused by PAN in MCs treated with it.[1]
- In cultured podocytes, Puromycin Aminonucleoside (10–50 μM) induced apoptosis characterized by caspase-3 activation, DNA fragmentation, and increased p53 expression. Western blot analysis revealed decreased Bcl-2 and increased Bax levels, indicating mitochondrial pathway involvement [2] - Treatment of mesangial cells with Puromycin Aminonucleoside (20–100 μM) led to apoptosis and inflammatory injury, marked by increased IL-6 and TNF-α secretion. This effect was mediated by estrogen-related receptor-α (ERRα), as ERRα knockdown using siRNA attenuated cytokine production [3] - Puromycin Aminonucleoside (50 μM) disrupted the actin cytoskeleton in podocytes, observed by reduced F-actin staining and increased nephrin phosphorylation, as detected by immunofluorescence and Western blot [6] |
ln Vivo |
The renal cortex of rats treated with PAN shows increased expression of ERRα, which is consistent with an enhanced apoptotic response, according to the in vitro study.[1]
- In rats, intravenous injection of Puromycin Aminonucleoside (100 mg/kg) induced nephrotic syndrome, characterized by proteinuria (peaking at 7 days post-injection), hypoalbuminemia, and renal histological changes including foot process effacement. Quantitative proteomics showed a 40% reduction in nephrin levels in podocytes [6] - In a mouse model of podocyte injury, systemic administration of Puromycin Aminonucleoside (150 mg/kg) increased urinary migrasomes, which correlated with early podocyte injury as detected by electron microscopy and flow cytometry [4] - Pretreatment of rats with an adenosine deaminase inhibitor (e.g., erythro-9-(2-hydroxy-3-nonyl)adenine, 5 mg/kg) before Puromycin Aminonucleoside (180 mg/kg) administration reduced proteinuria by 50% and preserved renal histology, indicating protection against nephrotoxicity [7] |
Cell Assay |
On 96-well plates, cells are seeded at a density of 5,000 cells per well in MEM containing 10% FBS. Following approximately 48 hours of incubation (approximately 40–50% confluence), cells are transferred to new growth medium that contains Puromycin aminonucleoside (NSC 3056) in different concentrations. In order to conduct the protection experiment, cells are cultured in a medium containing either 250 μM Puromycin aminonucleoside (NSC 3056) or 2 μM Decynium-22, a PMAT inhibitor. Following a 72-hour incubation period at 37°C in an incubator with 95% O2, the cells and plates are cleaned. The IC50 values are obtained by nonlinearly regressing the cell growth data to the following model (WinNonLin version 3.2): S is the percentage of optical density to untreated control cells' cell survival expressed as Smax − [Smax − S0] × [Cγ/(Cγ + IC50γ)], where S0 is the lowest residual cell survival at high drug concentrations, C is the concentration of Puromycin aminonucleosides, γ is the Hill coefficient, and IC50 is the concentration of Puromycin aminonucleosides that leads to half-maximal cell survival. In every experiment, four separate experiments are conducted, with five to six determinations made.
- Podocyte Apoptosis Assay: Human podocytes were treated with Puromycin Aminonucleoside (10–50 μM) for 48 hours. Apoptosis was measured by Annexin V/PI staining and flow cytometry, showing dose-dependent increases in apoptotic cells. Western blot analysis confirmed p53 upregulation and Bcl-2 downregulation [2] - Mesangial Cell Inflammation Assay: Rat mesangial cells were incubated with Puromycin Aminonucleoside (20–100 μM) for 24 hours. Culture supernatants were analyzed for IL-6 and TNF-α levels using ELISA, while cell lysates were assessed for ERRα expression by Western blot [3] - Nephrin Dynamics Assay: Mouse podocytes treated with Puromycin Aminonucleoside (50 μM) for 24 hours were subjected to quantitative proteomics. Selected reaction monitoring (SRM) mode detected a 30% decrease in nephrin phosphorylation at specific residues [6] |
Animal Protocol |
Male F344 rats at 11 weeks of age are purchased from JaPuromycin aminonucleoside SLC. In this study, a Puromycin aminonucleoside nephrosis model and normal rats are used. Rats are given a single intravenous injection of puromycin aminonucleoside (NSC 3056) at a dose of 8 mg/100 g body weight in saline to cause puromycin aminonucleoside nephrosis. The volume of saline given to control animals is the same. Days 4 and 7 following the Puromycin aminonucleoside injection are studied in nephrotic rats (n=6 per group).
- Nephrotic Syndrome Model in Rats: Male Sprague-Dawley rats (200–250 g) received a single intravenous injection of Puromycin Aminonucleoside (100 mg/kg dissolved in sterile saline). Urine was collected daily for protein measurement, and kidneys were harvested on days 3, 7, and 14 for histological and proteomic analysis [6] - Migrasome Detection in Mice: C57BL/6 mice (8–10 weeks old) were intraperitoneally injected with Puromycin Aminonucleoside (150 mg/kg). Urine samples collected at 24 and 48 hours were analyzed by electron microscopy and flow cytometry to quantify migrasomes [4] - Nephrotoxicity Protection Study in Rats: Animals received an adenosine deaminase inhibitor (5 mg/kg, intraperitoneal) 1 hour before Puromycin Aminonucleoside (180 mg/kg, intravenous). Proteinuria was monitored for 7 days, and renal tissue was evaluated for histological damage [7] |
Toxicity/Toxicokinetics |
- Acute Renal Toxicity: In rats, Puromycin Aminonucleoside (180 mg/kg) caused severe proteinuria (≥300 mg/24h) and histological changes including glomerular basement membrane thickening and foot process effacement [6]
- Protective Effect of Adenosine Deaminase Inhibition: Co-administration of an adenosine deaminase inhibitor reduced serum creatinine levels by 30% and preserved renal tubular structure compared to Puromycin Aminonucleoside alone [7] |
References |
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Additional Infomation |
3'-amino-3'-deoxy-N(6),N(6)-dimethyladenosine is puromycin derivative that lacks the methoxyphenylalanyl group on the amine of the sugar ring. It is a 3'-deoxyribonucleoside and a member of adenosines.
PUROMYCIN derivative that lacks the methoxyphenylalanyl group on the amine of the sugar ring. It is an antibiotic with antineoplastic properties and can cause nephrosis. - Puromycin Aminonucleoside-induced nephrotoxicity is characterized by podocyte injury, which is mediated by oxidative stress and mitochondrial dysfunction. The drug accumulates in podocytes via PMAT, leading to intracellular adenosine deaminase inhibition and ATP depletion [5] - The nephrotic syndrome model induced by Puromycin Aminonucleoside is widely used to study proteinuria and glomerular diseases, as it recapitulates human minimal change disease features. |
Molecular Formula |
C12H18N6O3
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Molecular Weight |
294.309721469879
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Exact Mass |
294.144
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Elemental Analysis |
C, 48.97; H, 6.16; N, 28.56; O, 16.31
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CAS # |
58-60-6
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Related CAS # |
58-60-6 (Puromycin Aminonucleoside); 58-58-2 (Puromycin 2HCl); 53-79-2 (Puromycin free base)
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PubChem CID |
6020
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Appearance |
White to light yellow solid powder
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Density |
1.7±0.1 g/cm3
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Boiling Point |
595.6±60.0 °C at 760 mmHg
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Melting Point |
235℃ (Decomposition)
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Flash Point |
314.0±32.9 °C
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Vapour Pressure |
0.0±1.8 mmHg at 25°C
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Index of Refraction |
1.776
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LogP |
0.02
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Hydrogen Bond Donor Count |
3
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Hydrogen Bond Acceptor Count |
8
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Rotatable Bond Count |
3
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Heavy Atom Count |
21
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Complexity |
373
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Defined Atom Stereocenter Count |
4
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SMILES |
OC[C@@H]1[C@@H](N)[C@H]([C@H](N2C=NC3=C2N=CN=C3N(C)C)O1)O
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InChi Key |
RYSMHWILUNYBFW-GRIPGOBMSA-N
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InChi Code |
InChI=1S/C12H18N6O3/c1-17(2)10-8-11(15-4-14-10)18(5-16-8)12-9(20)7(13)6(3-19)21-12/h4-7,9,12,19-20H,3,13H2,1-2H3/t6-,7-,9-,12-/m1/s1
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Chemical Name |
(2R,3R,4S,5S)-4-amino-2-[6-(dimethylamino)purin-9-yl]-5-(hydroxymethyl)oxolan-3-ol
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Synonyms |
NSC3056; NSC-3056; Puromycin Aminonucleoside; ARDMA; NSC 3056; SAN; Stylomycin aminonucleoside; Aminonucleoside; Stylomycin aminonucleoside; 3'-amino-3'-deoxy-n6,n6-dimethyladenosine; 3'-Amino-3'-deoxy-N,N-dimethyladenosine; 6-Dimethylamino-9-(3'-ribosylamine)purine; ADENOSINE, 3'-AMINO-3'-DEOXY-N,N-DIMETHYL-;
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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 |
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: 25~59 mg/mL (84.9~200.5 mM)
Water: ~30 mg/mL (~101.9 mM) |
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Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.49 mM) (saturation unknown) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (8.49 mM) (saturation unknown) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. 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: ≥ 2.08 mg/mL (7.07 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. Solubility in Formulation 4: ≥ 2.08 mg/mL (7.07 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 20.8 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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 5: ≥ 2.08 mg/mL (7.07 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 20.8 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 6: ≥ 2.08 mg/mL (7.07 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 20.8 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. Solubility in Formulation 7: ≥ 2.08 mg/mL (7.07 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 20.8 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. Solubility in Formulation 8: ≥ 2.08 mg/mL (7.07 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 9: ≥ 2.08 mg/mL (7.07 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL clear DMSO stock solution to 900 μL corn oil and mix evenly. Solubility in Formulation 10: 12.5 mg/mL (42.47 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
1 mM | 3.3978 mL | 16.9889 mL | 33.9778 mL | |
5 mM | 0.6796 mL | 3.3978 mL | 6.7956 mL | |
10 mM | 0.3398 mL | 1.6989 mL | 3.3978 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.