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| Other Sizes |
| Targets |
8-(p-Sulfophenyl)theophylline (8-SPT) targets the adenosine A2B receptor (A2BR), a G protein-coupled receptor (GPCR) that couples primarily to Gs proteins, activating adenylyl cyclase and increasing intracellular cAMP levels. The A2B receptor is the low-affinity adenosine receptor, activated by micromolar concentrations of adenosine, and is involved in inflammation, mast cell activation, vasodilation, and tumor growth. 8-SPT acts as a competitive antagonist at the A2B receptor, binding to the orthosteric adenosine binding site. It has a Ki value of 1330 nM (1.33 microM) for the human recombinant A2B receptor. 8-SPT is selective for A2BR over other adenosine receptor subtypes (A1, A2A, A3) at concentrations up to 10 microM.
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| ln Vitro |
In vitro, 8-(p-Sulfophenyl)theophylline (8-SPT) is a competitive antagonist of the adenosine A2B receptor. In functional assays using CHO cells expressing human A2B receptors, 8-SPT (1-100 microM) inhibits NECA (5‘-N-ethylcarboxamidoadenosine)-stimulated cAMP accumulation with an IC₅0 in the low micromolar range (approximately 2-5 microM). The calculated Ki value is 1330 nM. 8-SPT has no significant effect on baseline cAMP levels. The compound also inhibits A2B receptor-mediated relaxation of smooth muscle preparations (e.g., rat aorta) with an IC₅0 in the micromolar range. At concentrations up to 300 microM, 8-SPT does not significantly inhibit A1 or A2A receptors (selectivity >30-fold). The sulfophenyl group prevents cellular penetration, limiting its use to extracellular A2B receptors.
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| ln Vivo |
In vivo, 8-(p-Sulfophenyl)theophylline (8-SPT) has been used to study the role of A2B receptors in various disease models. In a mouse model of asthma (ovalbumin-induced airway inflammation), administration of 8-SPT (10-50 mg/kg, intraperitoneally) reduces airway hyperresponsiveness, eosinophil infiltration, and mucus production, confirming the pro-inflammatory role of A2B receptors. In a rat model of hindlimb ischemia, 8-SPT inhibits angiogenesis, indicating a role for A2B receptors in promoting blood vessel growth. In cancer models (e.g., melanoma, colon cancer), 8-SPT has been shown to reduce tumor growth and metastasis by blocking A2B receptor-mediated immunosuppression and promoting anti-tumor immunity. 8-SPT is also used as a reference antagonist to validate A2B receptor involvement in pharmacological responses.
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| Enzyme Assay |
In vitro A2B receptor binding assay: Membranes from CHO cells stably expressing human A2B receptors (20-50 ug protein) are incubated in assay buffer (50 mM Tris-HCl pH 7.4, 10 mM MgCl2, 1 mM EDTA, 0.1% BSA) with 1 nM [3H]-DPCPX (an adenosine antagonist) or [3H]-NECA, and varying concentrations of 8-(p-Sulfophenyl)theophylline (0.1-1000 uM) at 25degC for 60-90 min. Non-specific binding is determined with 10 uM NECA. Bound radioligand is separated by filtration through GF/B filters, and radioactivity is counted. The IC₅0 is calculated from competition curves, and the Ki is determined using the Cheng-Prusoff equation (Ki = 1330 nM for A2BR). For functional assays, CHO-A2B cells are seeded in 96-well plates, treated with 8-SPT (1-100 uM) for 30 min, then stimulated with NECA (10-100 nM). cAMP levels are measured by HTRF or ELISA. IC₅0 is calculated.
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| Cell Assay |
For cell-based A2B receptor functional assays, CHO cells expressing human A2B receptors are seeded in 96-well plates (2×10⁴ cells/well) in DMEM with 10% FBS and incubated overnight. Cells are washed with PBS and incubated with 8-(p-Sulfophenyl)theophylline (1-1000 uM) in assay buffer (PBS containing 0.5 mM IBMX, a phosphodiesterase inhibitor) for 15-30 min at 37degC. Then, NECA (10-100 nM) is added for 15-30 min to stimulate A2B receptor activation. The reaction is terminated by adding lysis buffer, and intracellular cAMP levels are quantified by ELISA or HTRF. The IC₅0 is calculated from the dose-response curve. For control experiments, vehicle (DMSO) alone and the potent A2B antagonist PSB-603 (1 nM-10 uM) are used. Cytotoxicity is assessed by MTT assay in parallel. 8-SPT is not toxic at concentrations up to 300 uM.
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| Animal Protocol |
In vivo mouse model of airway inflammation: Female BALB/c mice (6-8 weeks old, n=10/group) are sensitized with ovalbumin (OVA, 50 ug in alum) intraperitoneally on days 0 and 7. From day 14-21, mice are challenged with aerosolized OVA (1% in PBS) for 30 min daily. 8-(p-Sulfophenyl)theophylline (8-SPT) is formulated in 0.5% methylcellulose or saline and administered intraperitoneally (10-50 mg/kg) 30 min before each OVA challenge. Control groups receive vehicle alone or theophylline (50 mg/kg). At the endpoint (day 22), mice are euthanized, and bronchoalveolar lavage fluid (BALF) is collected for eosinophil count (by flow cytometry) and cytokine measurement (IL-4, IL-5, IL-13 by ELISA). Lung tissues are harvested for histology (H&E staining and PAS staining for mucus). 8-SPT treatment significantly reduces eosinophil infiltration and mucus production. Body weight is monitored for toxicity.
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| ADME/Pharmacokinetics |
8-(p-Sulfophenyl)theophylline (8-SPT, MW 336.33) is a hydrophilic, charged molecule due to the sulfonate group. It has poor oral bioavailability because it is not passively absorbed from the gastrointestinal tract. Following intraperitoneal or intravenous administration, 8-SPT distributes primarily in the extracellular fluid and does not readily cross the blood-brain barrier (BBB). The plasma half-life in rodents is approximately 1-2 hours. 8-SPT is not significantly metabolized and is excreted unchanged in the urine (renal clearance) due to its high water solubility. It does not accumulate in tissues. For in vivo studies, 8-SPT is typically administered intraperitoneally (10-50 mg/kg) or intravenously (5-20 mg/kg). For in vitro use, it is soluble in DMSO (50-100 mM) and water (10-20 mM). Storage: powder at -20degC, protected from light.
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| Toxicity/Toxicokinetics |
For 8-(p-Sulfophenyl)theophylline, hazard statements: H315 (Causes skin irritation), H319 (Causes serious eye irritation), H335 (May cause respiratory irritation). Signal word: Warning. Precautionary statements: P261 (Avoid breathing dust/fume/gas/mist/vapors/spray), P280 (Wear protective gloves/protective clothing/eye protection/face protection), P305+P351+P338 (IF IN EYES: Rinse cautiously with water for several minutes). The compound is not classified as a drug; it is a research chemical. Standard safety precautions for handling laboratory chemicals apply: use PPE (gloves, lab coat, goggles), work in a fume hood, avoid inhalation and skin contact. Storage: powder at -20degC for 3 years, 4degC for 2 years; in solvent at -80degC for 1 year, -20degC for 6 months. Protect from light.
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| References | |
| Additional Infomation |
adenosine antagonists
8-(p-Sulfophenyl)theophylline (8-SPT; 8-(4-Sulfophenyl)theophylline; CAS# 80206-91-3) is a research-grade adenosine A2B receptor antagonist (Ki = 1330 nM). It is not an FDA-approved drug. It is used as a pharmacological tool for studying the role of A2B receptors in inflammation, asthma, cardiovascular disease, and cancer. For research use only (RUO), not for diagnostic or therapeutic applications. |
| Molecular Formula |
C13H12N4O5S
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|---|---|
| Molecular Weight |
336.32
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| Exact Mass |
336.053
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| CAS # |
80206-91-3
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| PubChem CID |
1908
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| Appearance |
Solid powder
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| Hydrogen Bond Donor Count |
2
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
23
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| Complexity |
610
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CN1C2=C(C(=O)N(C1=O)C)NC(=N2)C3=CC=C(C=C3)S(=O)(=O)O
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| InChi Key |
LXJSJIXZOAMHTG-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C13H12N4O5S/c1-16-11-9(12(18)17(2)13(16)19)14-10(15-11)7-3-5-8(6-4-7)23(20,21)22/h3-6H,1-2H3,(H,14,15)(H,20,21,22)
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| Chemical Name |
4-(1,3-dimethyl-2,6-dioxo-7H-purin-8-yl)benzenesulfonic acid
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| Synonyms |
8-SPT; 8-pSPT
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.9734 mL | 14.8668 mL | 29.7336 mL | |
| 5 mM | 0.5947 mL | 2.9734 mL | 5.9467 mL | |
| 10 mM | 0.2973 mL | 1.4867 mL | 2.9734 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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