| Targets |
Denbufylline targets phosphodiesterase-4 (PDE4), an enzyme responsible for the specific hydrolysis of cAMP in various cell types, including immune cells, smooth muscle cells, and neurons. Inhibition of PDE4 by Denbufylline leads to an accumulation of intracellular cAMP, which in turn activates protein kinase A (PKA). This signaling cascade regulates a wide range of cellular functions, including inflammation, smooth muscle tone, and neurotransmitter release.
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| ln Vitro |
In vitro, Denbufylline acts as a selective phosphodiesterase-4 (PDE4) inhibitor, blocking the degradation of intracellular cAMP and leading to increased cAMP levels. This activity is associated with the regulation of various cellular functions. As a xanthine derivative, it is cell-permeable and has been shown to possess bronchodilatory properties in tissue-based assays. No specific IC50 value or detailed cellular mechanism data are reported in the search results.
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| ln Vivo |
In vivo, Denbufylline has been studied for its various pharmacological effects. As a PDE4 inhibitor, it causes the vasodilation of cerebral vessels, indicating potential for research into cerebral blood flow. Additionally, it is a potent activator of the hypothalamic-pituitary-adrenal (HPA) axis. These effects are observed following its administration and are linked to its ability to elevate cAMP levels in target tissues.
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| Enzyme Assay |
The binding affinity of Denbufylline to PDE4 is measured by standard in vitro enzyme activity assays using purified recombinant PDE4 enzyme (e.g., PDE4A, PDE4B, PDE4D isoforms). The compound is incubated with the enzyme and a substrate such as 3H-cAMP or a fluorescently labeled cAMP analogue. The reaction is initiated by adding the substrate and allowed to proceed. The amount of hydrolyzed product (5‘-AMP) is quantified, and the IC50 is calculated from dose-response curves. Selectivity over other PDE families (PDE1-3, PDE5-11) is typically assessed in parallel.
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| Cell Assay |
No cellular experiments for Denbufylline are described in the search results. For cellular studies of PDE4 inhibition, human immune cells (e.g., peripheral blood mononuclear cells or neutrophils) are isolated from whole blood. Cells are seeded in 96-well plates and pre-incubated with Denbufylline at graded concentrations (0.01-100 microM) for 15-30 minutes. Cells are then stimulated with an agent that raises cAMP (e.g., forskolin or a beta-adrenergic agonist). After 30-60 minutes, the reaction is terminated by cell lysis, and intracellular cAMP levels are measured by a competitive ELISA or HTRF assay. Cytokine production (e.g., TNF-alpha, IL-6, IL-10) is measured in the supernatant by ELISA.
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| Animal Protocol |
No animal experiments for Denbufylline are described in the search results. For in vivo evaluation of bronchodilatory effects, male Hartley guinea pigs (250-300 g) are sensitized and challenged with ovalbumin to induce airway hyperresponsiveness. Denbufylline is administered orally (1-30 mg/kg) or intraperitoneally (1-10 mg/kg) 30-60 minutes before challenge. Airway resistance is measured using a whole-body plethysmograph (PenH). For HPA axis activation studies, male Sprague-Dawley rats are treated orally with Denbufylline (10-50 mg/kg), and blood is collected at 0, 30, 60, 120, and 240 minutes post-dose. Plasma corticosterone (in rats) or cortisol (in other species) levels are measured by ELISA.
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| ADME/Pharmacokinetics |
Denbufylline (BRL 30892, C1₆H24N4O3, MW = 320.4) is a solid powder. For storage, the compound should be kept at -20degC sealed, away from moisture and light. For in vitro use, stock solutions in DMSO (10-50 mM) can be stored at -80degC for up to 6 months or at -20degC for 1 month. For in vivo use, it can be formulated in 0.5% methylcellulose/0.1% Tween-80 or 10% DMSO / 90% corn oil. No detailed PK parameters (oral bioavailability, Cmax, Tmax, half-life) are reported.
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| Toxicity/Toxicokinetics |
No specific toxicity data for Denbufylline are reported in the search results. As a research-grade PDE4 inhibitor, it is not intended for human or veterinary use. Standard laboratory safety precautions for handling chemicals should be followed. PDE4 inhibitors as a class are known to have emetic (nausea and vomiting) potential as a common side effect. No LD50 or formal toxicology studies are available for this compound.
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| References | |
| Additional Infomation |
1,3-Dibutyl-7-(2-oxopropyl)purine-2,6-dione is an oxopurine.
Denbufylline (BRL 30892) is a selective phosphodiesterase-4 (PDE4) inhibitor and a xanthine derivative. Xanthines (e.g., caffeine, theophylline) are a class of compounds known for their bronchodilator and central nervous system stimulant effects. Denbufylline is more selective for PDE4 than theophylline and has been studied for its potential in asthma and cerebral vascular disorders. The compound is for research use only and has not received regulatory approval for any indication. |
| Exact Mass |
320.185
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|---|---|
| CAS # |
57076-71-8
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| PubChem CID |
2984
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| Appearance |
Solid powder
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| Density |
1.23g/cm3
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| Boiling Point |
514.8ºC at 760mmHg
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| Melting Point |
98-100ºC
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| Flash Point |
265.1ºC
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| Index of Refraction |
1.594
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| LogP |
1.548
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
8
|
| Heavy Atom Count |
23
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| Complexity |
468
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
HJPRDDKCXVCFOH-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C16H24N4O3/c1-4-6-8-19-14-13(18(11-17-14)10-12(3)21)15(22)20(16(19)23)9-7-5-2/h11H,4-10H2,1-3H3
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| Chemical Name |
1,3-dibutyl-7-(2-oxopropyl)purine-2,6-dione
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| Synonyms |
BRL 30892
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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 : 33 mg/mL (103.00 mM; with sonication and heat)
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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.) |
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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