| Size | Price | Stock | Qty |
|---|---|---|---|
| 50mg |
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| 100mg |
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| 250mg | |||
| Other Sizes |
| Targets |
APRT (adenine phosphoribosyltransferase) – No IC50, Ki, or EC50 values were determined; the compound showed little irreversible or competitive inhibitory effect on APRT in vitro even at concentrations up to 1.5 × 10⁻¹ M [1]
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|---|---|
| ln Vitro |
Irreversible inhibition assay: Preincubation of ES cell extracts (E14.1) with 1.5 × 10⁻¹ M 9-ethyladenine for 1 hour resulted in a significant reduction of APRT activity to 0.01 ± 0.01 nmol/min/mg protein, compared to control (without 9-ethyladenine) which was 3.5 ± 0.1 nmol/min/mg protein. Lower concentrations (1.5 × 10⁻⁴ M, 1.5 × 10⁻³ M, 1.5 × 10⁻² M) did not cause significant inhibition (APRT activities ranged from 2.3 to 2.8 nmol/min/mg protein vs controls 2.7–3.0 nmol/min/mg protein) [1]
- Competitive inhibition assay: Even in the presence of 1.5 × 10⁻¹ M 9-ethyladenine, there was no significant reduction in [¹⁴C]-adenine incorporation into ES cell extracts compared to control [1] |
| ln Vivo |
Administration of 9-ethyladenine to HPRT-deficient mice (2.5 × 10⁻⁶ mol or 1.25 × 10⁻⁵ mol per injection, i.p., three times per week for 5 weeks) did not induce self-injurious behavior (no visible injury or hair loss) and did not significantly alter stereotypy frequency compared to wild-type controls [1]
- Brain APRT activity in mice treated with 2.5 × 10⁻⁶ mol 9-ethyladenine was 0.26 ± 0.04 nmol/min/mg protein vs saline control 0.25 ± 0.05; with 1.25 × 10⁻⁵ mol, APRT activity was 0.11 ± 0.02 vs saline control 0.12 ± 0.02 (no significant reduction) [1] |
| Enzyme Assay |
APRT activity measurement: Crude cell extracts were prepared by homogenizing cells or brain tissue in 0.03 M Tris-HCl (pH 7.4), followed by centrifugation at 12,000 rpm for 20 min at 4°C. The supernatant was used directly. The assay was performed as described by Meuth et al. Radiolabeled [¹⁴C]-adenine incorporation was measured, and labeled AMP was absorbed onto DE81 filters. Radioactivity was determined using a scintillation counter [1]
- Irreversible inhibition assay for APRT: Crude extract from E14.1 cells was preincubated with 9-ethyladenine (at concentrations ranging from 1.5 × 10⁻⁴ M to 1.5 × 10⁻¹ M) for 1 hour in a reaction mixture without [¹⁴C]-adenine. Then [¹⁴C]-adenine was added and incubation continued for another 1 hour. The labeled AMP was captured on DE81 filters and counted [1] - Competitive inhibition assay for APRT: Similar to the irreversible assay but without preincubation; 9-ethyladenine was added together with [¹⁴C]-adenine, and incorporation was measured [1] - HPRT activity measurement: Using [¹⁴C]-hypoxanthine as substrate, similar procedure as APRT assay [1] |
| Cell Assay |
ES cell culture: Wild-type E14.1 and HPRT-deficient E14.1TG3B1 cells were cultured in Dulbecco’s Modified Eagle Medium supplemented with 15% fetal calf serum, 0.1 mM β-mercaptoethanol, and 1000 U/mL leukemia inhibitory factor [1]
- Preparation of ES cell extracts: Cells (1–2 × 10⁷) grown to 80% confluency were trypsinized, washed twice with ice-cold PBS, and pelleted by centrifugation. The final pellet was resuspended in 3 mL of 0.03 M Tris-HCl (pH 7.4). Cells were homogenized using a Polytron, and the suspension was centrifuged at 12,000 rpm for 20 min at 4°C. The supernatant (crude extract) was used for enzyme assays. Protein concentration was determined by Bradford assay [1] |
| Animal Protocol |
Animals: Male C57BL/6 wild-type mice and HPRT-deficient male mice (C57/3B1, hybrid of 129/Ola and C57BL/6), 6–8 weeks old, were individually caged and maintained on a 12 h light/dark cycle [1]
- Dosing and administration: 9-Ethyladenine was dissolved in sterile normal saline at 2.5 × 10⁻² M and stored at 4°C. Mice received intraperitoneal injections of 2.5 × 10⁻⁶ mol or 1.25 × 10⁻⁵ mol of 9-ethyladenine three times per week for 5 weeks. A higher dose of 2.5 × 10⁻⁵ mol was also tested but caused lethality [1] - Behavioral observation: Before injection, mice were transferred to a clean cage with bedding but no food or water for 10 min. Recording started 10 min after returning the animals to the cage post-injection. Stereotypic behavior (grooming with fore or hind legs, nibbling, biting) was scored over a 20-min period. Recordings were made three times per week for 5 weeks (total of 15 times per mouse) [1] - Brain tissue collection: After the fourth injection, mice were sacrificed, brains were removed and homogenized to prepare cell extracts for APRT activity measurement [1] |
| Toxicity/Toxicokinetics |
At a dose of 1.25 × 10⁻⁵ mol of 9-ethyladenine (three times per week), after 4 weeks mice became too sick to move and one mouse died, likely due to chronic toxicity [1]
- At a higher dose of 2.5 × 10⁻⁵ mol (single intraperitoneal injection), 4 out of 5 male mice died after the first injection, and the remaining one died after the second injection, probably due to cytotoxicity [1] - No tissue injury or hair loss was observed in any animal within 120 days after starting injections, even at the doses tested [1] - Some mice treated with 9-ethyladenine showed increased sensitivity to sound and the presence of humans [1] |
| References | |
| Additional Infomation |
9-Ethyladenine was previously reported to be an efficient inhibitor of APRT and to cause self-injurious behavior (Lesch-Nyhan syndrome-like symptoms) in HPRT-deficient mice. However, the present study could not reproduce these findings [1]
- The study concluded that 9-ethyladenine is not a sufficient APRT inhibitor and cannot be used in experiments that mimic lowered APRT status in an animal model [1] |
| Molecular Formula |
C7H9N5
|
|---|---|
| Molecular Weight |
163.18
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| Exact Mass |
163.085
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| CAS # |
2715-68-6
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| PubChem CID |
7
|
| Appearance |
White to off-white solid powder
|
| Density |
1.5±0.1 g/cm3
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| Boiling Point |
382.2±45.0 °C at 760 mmHg
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| Melting Point |
196.5-197.5 °C(benzene)
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| Flash Point |
185.0±28.7 °C
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| Vapour Pressure |
0.0±0.9 mmHg at 25°C
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| Index of Refraction |
1.753
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| LogP |
0.26
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
12
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| Complexity |
162
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
MUIPLRMGAXZWSQ-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C7H9N5/c1-2-12-4-11-5-6(8)9-3-10-7(5)12/h3-4H,2H2,1H3,(H2,8,9,10)
|
| Chemical Name |
Adenine, 9-ethyl- (8CI)
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| Synonyms |
9-Ethyladenine NSC-14580 NSC14580NSC 14580
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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) |
H2O : ~100 mg/mL (~612.82 mM)
DMSO : ~62.5 mg/mL (~383.01 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 3 mg/mL (18.38 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 30.0 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: ≥ 3 mg/mL (18.38 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 30.0 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: ≥ 3 mg/mL (18.38 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 33.33 mg/mL (204.25 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 | 6.1282 mL | 30.6410 mL | 61.2820 mL | |
| 5 mM | 1.2256 mL | 6.1282 mL | 12.2564 mL | |
| 10 mM | 0.6128 mL | 3.0641 mL | 6.1282 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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