| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg |
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| 500mg |
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| Other Sizes |
| Targets |
Adenosine A2b receptor (in NIH 3T3 fibroblast membranes): Kβ = 2.3 ± 0.2 μM (n=3).
Adenosine A2a receptor (in PC12 cell membranes): Kβ = 20 μM (95% confidence limits 7.4–56 μM, n=3). The selectivity ratio (A2a Kβ / A2b Kβ) is approximately 9-fold in favor of the A2b receptor. [1] |
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| ln Vitro |
In PGT-β cells, alloxazine (0-30 μM, 20 minutes) suppresses the synthesis of cyclic AMP [1].
In membrane preparations, Alloxazine acted as an antagonist of NECA-elicited stimulation of adenylate cyclase. It exhibited a Kβ value of 2.3 ± 0.2 μM at the A2b receptor in NIH 3T3 fibroblast membranes, and a Kβ value of 20 μM (7.4–56 μM) at the A2a receptor in PC12 cell membranes. It was the only non-xanthine antagonist tested that showed selectivity for the A2b receptor over the A2a receptor, with a selectivity of about 9-fold. [1] |
| ln Vivo |
Alloxazine (1 µmol/L; dispersed across cortical surface for 0-20 minutes) reduces NECA-induced vasodilation [2].
Topical application of alloxazine (1 μmol/l) significantly suppressed the vasodilation of rat pial arteries induced by NECA (0.01-1 μmol/l). This was evidenced by a significantly increased EC25 value for NECA (from 0.02 ± 0.01 to 0.60 ± 0.04 μmol/l; mean dose ratio, 30; n=4; P<0.001). [2] Alloxazine (1 μmol/l) significantly inhibited the NECA (0.01 and 1 μmol/l)-stimulated release of nitrite/nitrate into the artificial cerebrospinal fluid suffusing over the cortical surface of rats (P<0.05). [2] Pretreatment with alloxazine (10 μmol/l, applied locally to the cranial window with a bolus volume of 100 μl for three times every 10 min) did not alter the lower limit of cerebral blood flow (CBF) autoregulation in rats during stepwise hypotension. The lower limit remained similar to the control group. [2] |
| Enzyme Assay |
Adenylate cyclase activity was measured in membrane preparations from NIH 3T3 fibroblasts and PC12 cells. The incubation mixture contained 0.1 mM [α-32P]ATP (0.9 μCi/tube for PC12 membranes, 2.7 μCi/tube for NIH 3T3 membranes), 10 μM GTP, 5 mM MgCl2, 0.1 mM cyclic AMP, 0.02 mg/mL adenosine deaminase, 0.1 mM rolipram, 0.2 mM EGTA, 5 U/tube creatine phosphokinase, 2.6 mM phosphocreatine, 30 μg/tube bovine serum albumin, and 50 mM Tris-HCl (pH 7.4) in a total volume of 250 μL. Alloxazine was added from stock solutions in water or DMSO (final DMSO concentration 4%, which did not affect PC12 membranes but increased basal activity in NIH 3T3 membranes by ~20%; a 4% DMSO control was included). The reaction was initiated by adding membrane protein (~10 μg for PC12, ~10–300 μg for NIH 3T3) and incubated for 10 min at 37°C. The reaction was stopped with 0.5 mL of 10% trichloroacetic acid, and 3H-cyclic AMP was added for recovery correction. Cyclic AMP was isolated by two-step chromatography using Dowex and alumina columns. Antagonist potency (Kβ) for Alloxazine was calculated using the Schild equation for cell membranes. [1]
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| Cell Assay |
Cell viability assay [1]
Cell Types: PGT-β Cell Tested Concentrations: 0-30 μM Incubation Duration: 20 minutes Experimental Results: Concentration-dependent inhibition of cyclic AMP production with IC50 of 2.9 μM. For the astrocyte study, acutely isolated astrocytes were used. The effect of alloxazine was assessed by its ability to prevent the increase in intracellular calcium concentration that is stimulated by NECA, a characteristic effect mediated by the A2B receptor in these cells. [2] |
| Animal Protocol |
Animal/Disease Models: Male SD (SD (Sprague-Dawley)) rat [2]
Doses: 1 µmol/L Route of Administration: Diffuse on the cortical surface; 1 µmol/L Primary Experimental Results:Dramatically inhibited vasodilation, EC25 value increased by 0.60 µmol/L. Male Sprague-Dawley rats (250-320 g) were anesthetized with urethan and prepared with a closed cranial window over the parietal cortex. For vasodilation experiments, the cortical surface was continuously suffused with artificial cerebrospinal fluid. Alloxazine (1 μmol/l) was applied by adding it to the artificial CSF for 30 minutes before and during the suffusion of the agonist (NECA). [2] For cerebral blood flow autoregulation experiments, after a craniotomy, a laser-Doppler flowmetry probe was placed ~0.2 mm above the cortical surface. Alloxazine (10 μmol/l) was applied locally to the open cranial window as a bolus volume of 100 μl, administered three times every 10 minutes. The lower limit of autoregulation was defined as the mean arterial blood pressure at which CBF decreased by 10% of its value at resting MABP. [2] |
| References |
[1]. Brackett LE, Daly JW. Functional characterization of the A2b adenosine receptor in NIH 3T3 fibroblasts. Biochem Pharmacol. 1994 Mar 2;47(5):801-14.
[2]. Shin HK, et al. Role of adenosine A(2B) receptors in vasodilation of rat pial artery and cerebral blood flow autoregulation. Am J Physiol Heart Circ Physiol. 2000 Feb;278(2):H339-44. |
| Additional Infomation |
Alloxazine is a benzo[g]pterin-2,4-dione. It is a tautomer of isoAlloxazine.
Alloxazine is a non-xanthine heterocyclic adenosine receptor antagonist. In this study, it was the only compound among several non-xanthine antagonists (including CP 66,713, HTQZ, tracazolate, CGS 15943A, HPPI, 9-methyladenine, N6-cyclohexyl-9-methyladenine, and 2-(2-phenylethoxy)-9-methyladenine) that exhibited selectivity for the A2b receptor over the A2a receptor. The selectivity ratio was approximately 9-fold, based on comparison of Kβ values in NIH 3T3 (A2b) versus PC12 (A2a) membranes. [1] |
| Molecular Formula |
C10H6N4O2
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|---|---|
| Molecular Weight |
214.180241107941
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| Exact Mass |
214.049
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| CAS # |
490-59-5
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| PubChem CID |
5372720
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.509g/cm3
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| Index of Refraction |
1.699
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| LogP |
0.159
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
16
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| Complexity |
333
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=CC=C2C(=C1)N=C3C(=N2)N=C(N=C3O)O
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| InChi Key |
HAUGRYOERYOXHX-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C10H6N4O2/c15-9-7-8(13-10(16)14-9)12-6-4-2-1-3-5(6)11-7/h1-4H,(H2,12,13,14,15,16)
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| Chemical Name |
1H-benzo[g]pteridine-2,4-dione
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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 : ~5 mg/mL (~23.34 mM)
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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 | 4.6690 mL | 23.3449 mL | 46.6897 mL | |
| 5 mM | 0.9338 mL | 4.6690 mL | 9.3379 mL | |
| 10 mM | 0.4669 mL | 2.3345 mL | 4.6690 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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