| Size | Price | Stock | Qty |
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| 250mg |
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| 500mg |
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| 1g |
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| 2g | |||
| Other Sizes |
Purity: ≥98%
Acefylline (also known as Theophylline-7-acetic acid and Theophyllineacetic acid), is an antagonist of the adenosine receptor and an activator of the peptidylarginine deiminase (PAD). It is a stimulant medication that is an analog of xanthine. In the pharmaceutical preparation etanautine, acefylline and diphenhydramine work together to counteract the drowsiness that diphenhydramine induces.
| Targets |
adenosine receptor
cAMP phosphodiesterase (PDE) isoenzymes (rat lung) ( theophylline-7-acetic acid: IC50 = 0.5 mmol/L for PDE I, 0.6 mmol/L for PDE II, 0.4 mmol/L for PDE III) [1] Peptidylarginine deiminases (PADs, including PAD1, PAD3) [2] |
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| ln Vitro |
In vitro activity: Acefylline (200 μM; 5-180 min) is a peptidylarginine deiminase (PAD) activator.[2] 1. Acefylline (theophylline-7-acetate) was evaluated for inhibitory effects on rat lung cAMP phosphodiesterase (PDE) isoenzymes (PDE I, II, III); it showed weaker inhibitory activity than theophylline and ambroxol-theophylline-7-acetate, and comparable activity to ambroxol; theophylline-7-acetic acid (parent structure of acefylline) exhibited concentration-dependent inhibition of PDE I (IC50=0.5 mmol/L), PDE II (IC50=0.6 mmol/L), and PDE III (IC50=0.4 mmol/L) in rat lung tissue, with higher potency than ambroxol and lower potency than theophylline. [1] 2. Acefylline (100 μM) significantly activated filaggrin deimination in human epidermal keratinocytes (NHEK) and reconstructed human epidermis models (p < 0.05 vs control); it upregulated PAD1 and PAD3 protein expression in the upper epidermis (immunofluorescence staining, confocal microscopy), and increased citrullinated filaggrin levels (western blot with anti-citrulline antibody); the activation effect was dose-dependent (50/100/200 μM, peak at 100 μM, 200 μM showed no further increase); it had no effect on PAD2/PAD4 expression or localization in the epidermis. [2] |
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| ln Vivo |
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| Enzyme Assay |
1. Rat lung cAMP PDE isoenzyme inhibition assay: Rat lung tissue homogenates were prepared to isolate cAMP phosphodiesterase isoenzymes (PDE I, II, III); the enzyme reaction system was established with specific substrates and cofactors for each PDE isoenzyme; acefylline, theophylline, theophylline-7-acetic acid, ambroxol, and ambroxol-theophylline-7-acetate were added at different concentrations (0.1–1.0 mmol/L); the hydrolysis of cAMP was quantified to calculate inhibitory rates and IC50 values for each compound; the inhibitory potency of acefylline was compared with other test compounds (theophylline > theophylline-7-acetic acid > ambroxol-theophylline-7-acetate ≈ ambroxol ≈ acefylline). [1]
2. PAD activity assay for filaggrin deimination: Reconstructed human epidermis models and NHEK cells were treated with acefylline at 50/100/200 μM for 48 h; cell/tissue lysates were prepared, and PAD enzyme activity was measured using a fluorometric assay with specific substrates for PAD1/PAD3; citrullinated filaggrin levels were detected by western blot (anti-citrulline antibody) to confirm PAD-mediated deimination; immunofluorescence staining was performed to localize PAD1/PAD3 and citrullinated filaggrin in the upper epidermis (confocal microscopy, image quantification). [2] |
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| Cell Assay |
1. Human epidermal keratinocyte (NHEK) assay: NHEK cells were cultured in complete keratinocyte medium and seeded in 6-well plates; cells were treated with acefylline at 50/100/200 μM or vehicle (DMSO) for 48 h; total protein extracts were prepared, and western blot was performed to detect PAD1, PAD3, PAD2, PAD4, and citrullinated filaggrin levels (β-actin as loading control); immunofluorescence staining was conducted on fixed cells to visualize the subcellular localization of PAD1/PAD3 and citrullinated filaggrin (confocal microscopy, 3 independent experiments, one-way ANOVA for statistical analysis). [2]
2. Reconstructed human epidermis assay: 3D reconstructed human epidermis models were cultured in maintenance medium and treated with acefylline at 50/100/200 μM for 48 h; tissues were fixed, embedded in paraffin, and sectioned; hematoxylin-eosin (H&E) staining was performed to assess epidermal structure; immunofluorescence staining was used to detect PAD1/PAD3 expression and citrullinated filaggrin distribution in the upper epidermis (confocal microscopy, image analysis with ImageJ software, p < 0.05 vs control). [2] |
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| Animal Protocol |
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| References |
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| Additional Infomation |
2-(1,3-Dimethyl-2,6-dioxo-7-purine)acetic acid is an oxopurine. Acetyltheophylline has been reported in Pogostemon cablin, and relevant data are available. 1. Acetyltheophylline (7-acetyltheophylline) is a theophylline derivative with cAMP phosphodiesterase (PDE) inhibitory activity; its parent compound, 7-acetyltheophylline, is a selective inhibitor of rat lung cAMP PDE isoenzymes (PDE I > III > II), and its PDE inhibitory efficacy can be modulated by esterification/derivation (e.g., with ambroxol); the PDE inhibitory efficacy of acetyltheophylline is lower than that of theophylline but comparable to that of ambroxol. [1]
2. Acetylcillin is a novel peptidyl arginine deiminase (PAD1/PAD3) activator that acts on the epidermis to promote the deiminization (citrullination) of filaggrin—a key step in epidermal barrier formation and hydration; it has potential application value in treating dry skin diseases (e.g., atopic dermatitis) by enhancing the processing of epidermal filaggrin; it specifically targets PAD1/PAD3 without affecting other PAD subtypes (PAD2/PAD4), thereby reducing off-target effects. [2] 3. Acetylcillin belongs to the 7-theophylline acetate derivatives; [3] A method for synthesizing 7-theophylline acetate esters and diethylene glycol monomethyl ethers (structural analogs of acetylcillin) by esterification (concentrated sulfuric acid catalysis, reflux conditions) is described, aiming to improve their solubility and pharmacological properties (these analogs were not evaluated in vitro/in vivo). [3] |
| Molecular Formula |
C9H10N4O4
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| Molecular Weight |
238.2
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| Exact Mass |
238.07
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| Elemental Analysis |
C, 45.38; H, 4.23; N, 23.52; O, 26.87
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| CAS # |
652-37-9
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| Related CAS # |
18833-13-1 (piperazine); 652-37-9
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| PubChem CID |
69550
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| Appearance |
White to off-white solid powder
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| Density |
1.6±0.1 g/cm3
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| Boiling Point |
555.7±56.0 °C at 760 mmHg
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| Melting Point |
270-272 °C
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| Flash Point |
289.9±31.8 °C
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| Vapour Pressure |
0.0±1.6 mmHg at 25°C
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| Index of Refraction |
1.714
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| LogP |
-0.49
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
17
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| Complexity |
385
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C1C2=C(N=C([H])N2C([H])([H])C(=O)O[H])N(C([H])([H])[H])C(N1C([H])([H])[H])=O
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| InChi Key |
HCYFGRCYSCXKNQ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C9H10N4O4/c1-11-7-6(8(16)12(2)9(11)17)13(4-10-7)3-5(14)15/h4H,3H2,1-2H3,(H,14,15)
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| Chemical Name |
2-(1,3-dimethyl-2,6-dioxopurin-7-yl)acetic acid
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| Synonyms |
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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 |
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| 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) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (10.50 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 25.0 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: ≥ 2.5 mg/mL (10.50 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.1982 mL | 20.9908 mL | 41.9815 mL | |
| 5 mM | 0.8396 mL | 4.1982 mL | 8.3963 mL | |
| 10 mM | 0.4198 mL | 2.0991 mL | 4.1982 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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