| Size | Price | Stock | Qty |
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| Targets |
Benzodiazepine receptors (GABAA receptor benzodiazepine binding site). Ro 15-4513 is a high-affinity benzodiazepine ligand with Ki values of 3.1 nM for diazepam-insensitive (DI) and 5.3 nM for diazepam-sensitive (DS) benzodiazepine receptors. It acts as a partial inverse agonist at benzodiazepine receptors and has agonist activity at GABAA receptors containing α4 and α6 subunits.
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| ln Vitro |
For GABAA receptors, Ro15-4513 typically functions as a partial inverse agonist, with the exception of agonists that contain α4 and α6 subunits [3].
In vitro, Ro 15-4513 demonstrates high-affinity binding to benzodiazepine receptors with Ki values of 3.1 nM for diazepam-insensitive receptors and 5.3 nM for diazepam-sensitive receptors. The compound's partial inverse agonist activity at benzodiazepine receptors results in effects opposite to those of benzodiazepine agonists, including anxiogenic and proconvulsant effects. Its agonist activity at GABAA receptors containing α4 and α6 subunits is notable. |
| ln Vivo |
Complete inhibition of the ethanol (1.8 g/kg)-induced decrease in overall locomotor activity and partial inhibition of the ethanol-induced drop in backup force were observed with Ro 15-4513 (ip; 3 mg/kg; 10 min before testing) [2]. In mice lacking the GABAA receptor delta subunit, Ro 15-4513 (ip injection; 3 mg/kg; 15 minutes prior to 1.5 g/kg ethanol administration) reverses ethanol-induced drowsiness [2].
In vivo, Ro 15-4513 is an effective ethanol antagonist. It blocks many of the behavioral effects of ethanol, including its anxiolytic and motor-impairing effects. The compound's ability to antagonize ethanol effects is related to its activity at GABAA receptors containing α4 and α6 subunits. Ro 15-4513 has been extensively used as a research tool to study the role of GABAA receptors in ethanol action and addiction. |
| Enzyme Assay |
Cell-free receptor binding assays for Ro 15-4513 use membrane preparations from brain tissue or from cells expressing recombinant GABAA receptor subunits. The compound is incubated with a radiolabeled benzodiazepine ligand (e.g., [³H]-flumazenil or [³H]-diazepam) at varying concentrations for 60-120 minutes at 4°C. Nonspecific binding is determined in the presence of an excess of unlabeled benzodiazepine. Bound and free radioactivity are separated by filtration, and Ki values are calculated from competition curves.
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| Cell Assay |
Cellular functional assays for Ro 15-4513 use cells expressing recombinant GABAA receptors with specific subunit compositions (e.g., α1β2γ2, α4β2γ2, or α6β2γ2). Cells are transfected with the appropriate GABAA receptor subunits and loaded with a fluorescent calcium indicator or voltage-sensitive dye. Cells are treated with GABA and Ro 15-4513 at varying concentrations, and receptor modulation is measured by assessing chloride flux using patch-clamp electrophysiology or fluorescence-based membrane potential assays. The compound's inverse agonist or agonist activity is determined based on its effects on GABA-induced currents.
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| Animal Protocol |
Animal/Disease Models: Male C57BL/6J mice [2]
Doses: 3 mg/kg Route of Administration: intraperitoneal (ip) injection; 10 minutes before test Experimental Results: Complete inhibition of ethanol-induced decrease in total locomotor activity and partial decrease in standing. In vivo studies with Ro 15-4513 are typically performed in rodents to assess its effects on ethanol-induced behaviors. The compound is administered intraperitoneally at doses typically ranging from 0.1-10 mg/kg. Behavioral assays such as the elevated plus maze, open field test, or rotarod are used to assess anxiolytic, motor, and sedative effects. Ethanol antagonism is assessed by co-administering ethanol and measuring the reversal of ethanol-induced effects. Brain penetration and receptor occupancy can be confirmed by ex vivo binding studies. |
| ADME/Pharmacokinetics |
Pharmacokinetic studies of Ro 15-4513 demonstrate that the compound is brain-penetrant, consistent with its activity as a benzodiazepine receptor ligand. PK parameters such as Cmax, Tmax, AUC, half-life, and brain-to-plasma ratio are determined in preclinical species. The compound's rapid brain penetration and receptor binding kinetics are consistent with its use as a research tool for studying GABAA receptor function.
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| Toxicity/Toxicokinetics |
Toxicological data for Ro 15-4513 are limited to research studies. As a benzodiazepine receptor inverse agonist, the compound can produce anxiogenic and proconvulsant effects at higher doses. Standard toxicology studies would be required for clinical development. The compound is handled with standard laboratory precautions for neuroactive compounds.
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| References |
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| Additional Infomation |
8-Azide-5-methyl-6-oxo-4H-imidazo[1,5-a][1,4]benzodiazepine-3-carboxylic acid ethyl ester is an organic nitrogen heterocyclic compound and an organic heterotricyclic compound.
Ro 15-4513 is a well-characterized research tool for studying GABAA receptor pharmacology and ethanol action. It was developed by Hoffmann-La Roche in 1984 and is structurally related to flumazenil. The compound is not an approved therapeutic agent and is available for research purposes only. Its unique pharmacological profile, including activity at diazepam-insensitive receptors and GABAA receptors containing α4 and α6 subunits, makes it a valuable tool for dissecting the roles of different GABAA receptor subtypes. |
| Molecular Formula |
C16H17N7O3
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|---|---|
| Molecular Weight |
355.36
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| Exact Mass |
326.113
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| CAS # |
91917-65-6
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| PubChem CID |
5081
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| Appearance |
Off-white to yellow solid powder
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| LogP |
1.967
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
24
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| Complexity |
563
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
CFSOJZTUTOQNIA-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C15H14N6O3/c1-3-24-15(23)13-12-7-20(2)14(22)10-6-9(18-19-16)4-5-11(10)21(12)8-17-13/h4-6,8H,3,7H2,1-2H3
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| Chemical Name |
ethyl 8-azido-5-methyl-6-oxo-4H-imidazo[1,5-a][1,4]benzodiazepine-3-carboxylate
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| Synonyms |
Ro-15-4513; Ro 154513; Ro 15-4513
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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 : ~10 mg/mL (~30.65 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1 mg/mL (3.06 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 10.0 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.8140 mL | 14.0702 mL | 28.1405 mL | |
| 5 mM | 0.5628 mL | 2.8140 mL | 5.6281 mL | |
| 10 mM | 0.2814 mL | 1.4070 mL | 2.8140 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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