Doxefazepam is a benzodiazepine analogue with anxiolytic (anti-anxiety) and anticonvulsant (antiepileptic/antiseizure) effects.

Physicochemical Properties

Molecular Formula	C17H14CLFN2O3
Molecular Weight	348.76
Exact Mass	348.068
CAS #	40762-15-0
PubChem CID	38668
Appearance	Typically exists as solid at room temperature
Density	1.45g/cm3
Boiling Point	618.4ºC at 760mmHg
Melting Point	138-140°
Flash Point	327.8ºC
Index of Refraction	1.646
LogP	1.474
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	3
Heavy Atom Count	24
Complexity	504
Defined Atom Stereocenter Count	0
SMILES	FC1C(C2C3C=C(Cl)C=CC=3N(CCO)C(=O)C(O)N=2)=CC=CC=1
InChi Key	VOJLELRQLPENHL-UHFFFAOYSA-N
InChi Code	InChI=1S/C17H14ClFN2O3/c18-10-5-6-14-12(9-10)15(11-3-1-2-4-13(11)19)20-16(23)17(24)21(14)7-8-22/h1-6,9,16,22-23H,7-8H2
Chemical Name	7-chloro-5-(2-fluorophenyl)-3-hydroxy-1-(2-hydroxyethyl)-3H-1,4-benzodiazepin-2-one
HS Tariff Code	2934.99.9001
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)

Biological Activity

Targets	Benzodiazepine[1]
ln Vivo	Food containing enough doxefazepam, a benzodiazepine derivative, is fed to groups of 50 male and 50 female Sprague-Dawley rats so that intakes of 0, 3, 10, or 30 mg/kg/day are guaranteed. After receiving treatment for 104 weeks, rats are put to death. Both euthanized animals and those who died simultaneously undergo a thorough autopsy. The rodents' ability to survive is unaffected by the long-term injection of doxefazepam. In the treated female groups, there is a noteworthy upward trend in the incidence of hepatocellular neoplasms, mostly benign ones. In treated rats, this increased prevalence is not linked to an increased incidence of focal hyperplasia or other preneoplastic lesions. Doxefazepam's pharmacological action targets the brain, which is why it is investigated thoroughly to look for tiny foci of proliferating cells. In male and female rats, a total of 12 and 6 malignant gliomas are found; only 2 are identified upon autopsy. The majority of these tumors are oligodendrogliomas, which are frequently observed in elderly rats [1]. Doxefazepam is the subject of several toxicological investigations. In mice, rats, and dogs, oral LD50 values exceed 2000 mg/kg, but endoperitoneal LD50 values in mice and rats are 746 and 544 mg/kg, respectively, and more than 1000 mg/kg in dogs[2].
Toxicity/Toxicokinetics	Toxicity Summary Benzodiazepines bind nonspecifically to benzodiazepine receptors BNZ1, which mediates sleep, and BNZ2, which affects affects muscle relaxation, anticonvulsant activity, motor coordination, and memory. As benzodiazepine receptors are thought to be coupled to gamma-aminobutyric acid-A (GABAA) receptors, this enhances the effects of GABA by increasing GABA affinity for the GABA receptor. Binding of the inhibitory neurotransmitter GABA to the site opens the chloride channel, resulting in a hyperpolarized cell membrane that prevents further excitation of the cell.
References	[1]. Borelli G, et al. Carcinogenicity study of doxefazepam administered in the diet to Sprague-Dawley rats. Fundam Appl Toxicol. 1990 Jul;15(1):82-92. [2]. Bertoli D, et al. Toxicological evaluations of the benzodiazepine doxefazepam. Arzneimittelforschung. 1989 Apr;39(4):480-4.
Additional Infomation	Doxefazepam is a benzodiazepine. Doxefazepam (marketed under brand name Doxans) is a benzodiazepine derivative drug developed by Schiapparelli in the 1970s. It possesses anxiolytic, anticonvulsant, sedative and skeletal muscle relaxant properties. It is used therapeutically as a hypnotic. According to Babbini and colleagues in 1975, this derivative of flurazepam was between 2 and 4 times more potent than the latter while at the same time being half as toxic in laboratory animals.

Solubility Data

Solubility (In Vitro)

DMSO: 250 mg/mL (716.83 mM)

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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.8673 mL	14.3365 mL	28.6730 mL
	5 mM	0.5735 mL	2.8673 mL	5.7346 mL
	10 mM	0.2867 mL	1.4337 mL	2.8673 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.