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| 1mg |
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| Other Sizes |
| Targets |
GABA receptor channels (GABA RCs). Gidazepam, the unlabeled parent compound, is an agonist of GABA type A (GABAA) receptor chloride channels. By binding to benzodiazepine binding sites on the GABAA receptor complex, Gidazepam enhances the affinity of the receptor for GABA, potentiating GABA-induced chloride ion influx. This leads to hyperpolarization of neurons and decreased neuronal excitability, resulting in anxiolytic, sedative, muscle relaxant, and anticonvulsant effects. Gidazepam-d5 is chemically identical to Gidazepam except for the substitution of five hydrogen atoms with deuterium atoms, which does not affect the molecular mechanism or binding affinity but provides a stable isotope label for analytical quantitation. The anticonvulsant effect of Gidazepam is utilized in preclinical epilepsy research.
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| ln Vitro |
No specific in vitro activity data are available for Gidazepam-d5. As a stable isotope-labeled analog, it is expected to have identical pharmacological properties to the parent drug Gidazepam. Gidazepam itself acts as a GABA receptor channel agonist, enhancing GABA-induced Cl- influx in GABAA receptor-expressing cells (e.g., Xenopus oocytes expressing specific GABAA receptor subunits or neuronal cell cultures). The compound potentiates the effect of GABA in a concentration-dependent manner, shifting the GABA dose-response curve to the left. In radioligand binding assays, Gidazepam displaces [3H]-flunitrazepam from the benzodiazepine binding site on the GABAA receptor, with affinity similar to classical benzodiazepines. Gidazepam-d5 is used as an internal standard for such binding assays in mass spectrometry-based workflows.
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| ln Vivo |
No specific in vivo data are available for Gidazepam-d5. The unlabeled parent compound Gidazepam exerts anticonvulsant effects in animal models of epilepsy, including pentylenetetrazole (PTZ)-induced seizures, maximal electroshock (MES) seizure models, and kindling models. In these models, Gidazepam elevates seizure threshold, reduces seizure severity and duration, and prevents seizure generalization. The compound also displays anxiolytic and sedative effects at higher doses. Gidazepam-d5 is typically not used for in vivo efficacy studies but rather as a tracer or internal standard in pharmacokinetic or toxicokinetic studies where deuterium labeling facilitates LC-MS/MS quantification of parent drug concentrations in plasma, brain, and other tissues.
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| Enzyme Assay |
For non-cellular binding assays, a radioligand binding assay can be performed using rat brain cortical or hippocampal membranes as a source of GABAA receptors. Membranes (100-200 ug protein) are prepared by homogenization in ice-cold Tris-citrate buffer (50 mM, pH 7.4) and centrifugation (20,000 × g, 20 min, 4degC). The pellet is washed and resuspended. For displacement assays, membranes are incubated with 0.5-1 nM [3H]-flunitrazepam (a benzodiazepine site radioligand) and varying concentrations of unlabeled Gidazepam (0.1-1000 nM) or a fixed concentration of Gidazepam-d5 (for isotope dilution experiments) in Tris-citrate buffer containing 150 mM NaCl for 60-90 minutes at 4degC. Non-specific binding is determined in the presence of 10 uM diazepam. Bound and free radioligand are separated by rapid filtration through GF/B filters pre-soaked in 0.3% polyethyleneimine using a cell harvester. Filters are washed, and radioactivity is quantified by liquid scintillation counting. Gidazepam-d5 can also be used as a competitor in this assay.
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| Cell Assay |
Gidazepam-d5 is not typically used in cell-based pharmacological assays because the deuterium label does not alter biological activity. For the unlabeled parent compound Gidazepam, a standard in vitro assay uses HEK293 cells transiently or stably transfected with GABAA receptor subunits (e.g., alpha1beta2gamma2). Cells are seeded in 96-well plates (2-4 × 10^4 cells/well) in DMEM with 10% FBS and incubated for 24-48 hours at 37degC, 5% CO2. For functional assessment of GABAA receptor activation, a fluorescent membrane potential assay or calcium-sensitive dye (e.g., Fluo-4 AM) can be used because GABAA receptor activation leads to Cl- influx, which can be measured indirectly. Alternatively, a patch-clamp electrophysiology assay is the gold standard: cells are voltage-clamped at -60 mV, and GABA (1-10 uM) is applied in the absence or presence of varying concentrations of Gidazepam (0.1-1000 nM). The potentiation of GABA-induced Cl- current is measured. Gidazepam-d5 can be used as an internal standard in cell lysates for LC-MS/MS analysis of drug concentrations in cellular uptake or metabolism studies.
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| Animal Protocol |
For in vivo pharmacokinetic studies, male Sprague-Dawley rats (250-300 g) or CD-1 mice (25-30 g) are used (n=3-5 per time point). Gidazepam-d5 (the parent compound or a mixture of unlabeled Gidazepam with Gidazepam-d5 as internal standard) is formulated in a suitable vehicle (e.g., 5% DMSO, 5% Kolliphor EL, 90% saline) and administered intravenously (0.5-2 mg/kg) or orally (2-10 mg/kg) by gavage. Blood samples (100-200 uL) are collected from the tail vein or via jugular vein cannulation into EDTA-coated tubes at pre-dose, 5, 15, 30, 60, 120, 240, 480, 720, and 1440 minutes post-dose. Plasma is separated by centrifugation (2,000 g, 10 min, 4degC) and stored at -80degC. At terminal time points, animals are euthanized, and tissues (brain, liver, kidney, heart, lung) are harvested, homogenized in PBS or water, and stored at -80degC. Gidazepam and Gidazepam-d5 concentrations are quantified by LC-MS/MS following protein precipitation with acetonitrile (3:1 v/v). PK parameters (AUC, Cmax, Tmax, t1/2, CL, Vd, bioavailability) are calculated by non-compartmental analysis using software (e.g., Phoenix WinNonlin).
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| ADME/Pharmacokinetics |
Gidazepam pharmacokinetics: As a benzodiazepine derivative, Gidazepam is highly lipophilic, with high plasma protein binding (>90% bound to albumin and alpha-1-acid glycoprotein). The volume of distribution is large (>2 L/kg), indicating extensive tissue distribution, particularly to the brain (due to its ability to cross the blood-brain barrier). The plasma half-life of Gidazepam in rodents is 2-6 hours, depending on the species and route of administration. The compound undergoes hepatic metabolism primarily via CYP3A4-mediated N-demethylation and aliphatic hydroxylation, followed by glucuronidation. Metabolites are excreted in urine and feces. Gidazepam-d5, being an isotopologue, has identical PK properties to the unlabeled compound, making it an ideal internal standard for bioanalysis. The deuterium atoms may slightly alter the rate of metabolism (kinetic isotope effect) in some cases, but this is generally minimal and does not significantly impact quantitation.
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| Toxicity/Toxicokinetics |
No specific toxicity data are available for Gidazepam-d5. The unlabeled parent Gidazepam has a safety profile consistent with classical benzodiazepines. Acute toxicity (high doses) results in CNS depression, sedation, ataxia, muscle relaxation, and respiratory depression, with lethal dose exceeding 100-200 mg/kg in rodents. With chronic administration, tolerance may develop, and physical dependence can occur, with withdrawal symptoms (anxiety, seizures, insomnia) upon discontinuation. No specific genotoxicity, carcinogenicity, or organ toxicity has been reported for Gidazepam alone. The deuterium label does not introduce additional toxicity. Gidazepam-d5 is a research chemical and is not intended for human or therapeutic use. Standard laboratory safety precautions for handling benzodiazepines (e.g., use of gloves, fume hood, proper disposal) should be followed.
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| References |
[1]. N. Ya Golovenko, et al. Pharmacodynamical and Neuroreceptor Analysis of the Permeability of the Blood-Brain Barrier for Derivatives of 1,4-Benzodiazepine. Neurophysiology, Vol. 46, No. 3, June, 2014.
[2]. Nesterkina M, et al. Synthesis and Pharmacological Properties of Novel Esters Based on Monocyclic Terpenes and GABA. Pharmaceuticals (Basel). 2016 Jun 13;9(2). pii: E32. |
| Additional Infomation |
Deuterium labeling (stable isotope labeling) is a common technique in pharmaceutical research to generate internal standards for quantitative mass spectrometry (LC-MS/MS). Gidazepam-d5 contains five deuterium atoms, providing a mass shift of +5 Da compared to the unlabeled parent Gidazepam. This mass difference allows the labeled compound to be distinguished from the unlabeled drug in biological samples, making it an ideal internal standard to account for matrix effects, extraction recovery, and ionization efficiency in bioanalysis. Gidazepam (unlabeled) is not an FDA-approved drug but is a research benzodiazepine known to have anticonvulsant properties. Gidazepam-d5 is for research use only; it is not a therapeutic agent. The compound is typically supplied as a solid powder and is stable for storage at -20degC protected from light.
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| Molecular Formula |
C17H10D5BRN4O2
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| Molecular Weight |
392.26
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| Related CAS # |
Gidazepam;129186-29-4
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| Appearance |
Typically exists as solid at room temperature
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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 | 2.5493 mL | 12.7466 mL | 25.4933 mL | |
| 5 mM | 0.5099 mL | 2.5493 mL | 5.0987 mL | |
| 10 mM | 0.2549 mL | 1.2747 mL | 2.5493 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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