Drug molecules have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as tracers for quantitation during the drug development process. Due to its potential to alter the pharmacokinetic and metabolic profiles of medications, deuteration has drawn attention[1].

Absorption, Distribution and Excretion
/MILK/ In goats, 1 hr after oral administration of 30 mg/kg bw nitrosodiethylamine, there were 11.4 mg/kg nitrosodiethylamine in milk and 11.9 mg/kg in blood. Only traces were found in milk and none in blood after 24 hr.
Autoradiographic studies indicated that non-metabolized N-nitrosodiethylamine passed to fetuses with even distribution in most fetal tissues on all studied days of gestation (day 12, 14, 16, 16 and 18) in mice. Results also indicated metabolism of the substance in mucosa of fetal bronchial tree and liver on day 18 of gestation.

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Metabolism / Metabolites
Inhibition of sulfotransferase by 2,6-dichloro-4-nitrophenol completely abolished the genotoxic potential of N-nitrosodiethanolamine in rat liver as indicated by the induction of DNA single-strand breaks. The DNA strand-breaking potential of N-nitroso-2-hydroxymorpholine, a metabolite of N-nitrosodiethanolamine formed by alcohol dehydrogenase -mediated oxidation, was also almost quantitatively abolished. In contrast to these beta-hydroxylated nitrosamines, the effectiveness of N-nitrosodiethylamine remained unaffected by 2,6-dichloro-4-nitrophenol with respect to its DNA damaging potential. ... A new activation mechanism for N-nitrosodiethanolamine is proposed: N-nitrosodiethanolamine is transformed at first by alcohol dehydrogenase into the cyclic hemiacetal N-nitroso-2-hydroxymorpholine. This cyclic beta-hydroxynitrosamine appears to be a substrate for sulfotransferase. The resulting sulfate conjugate is suggested to be ultimate genotoxic electrophile. However, the results do not exclude the possibility that N-nitrosodiethanolamine itself undergoes sulfate conjugation.
Oxidative N-deethylation of NDEA accounts for the production of CO2 and alkylating species in vivo. The rate of metabolism of NDEA by slices of organs from rats and hamsters in vitro has been measured, and a correlation made between the degree of metabolism and the distribution of induced tumors. After administration of NDEA to rats or hamsters, several ethylated derivatives were produced in liver and kidney nucleic acids. These included 7-ethylguanine, O6-ethylguanine and 3-ethyladenine.
... Evidence suggests that nitrosodiethylamine requires metabolic activation in order to exert its carcinogenic and toxic effects. ... N-nitrosoethyl-N-(2-hydroxyethyl)amine and N-nitrosoethyl-N-(carboxymethyl)amine have been detected in urine of rats. ...
Possible relationships between structure and metabolism of nitrosamines have been investigated in the rat small intestine. Isolated segments of jejunum and ileum were perfused from the luminal side for 2 hr with a Tyrode solution containing one of four symmetrical dialkylnitrosamines with 2-5 carbon atoms per side chain, all (14)C-labeled at the alpha position, or one of two unsymmetrical nitrosamines, N-nitroso-tert-butylmethylamine and N-nitrosomethylbenzylamine, (14)C-labeled in the methyl group. Besides measurement of (14)C to intestinal tissue, the absorbed fluid (absorbate) as well as the perfusion medium and tissue homogenates were analyzed by for the presence of polar metabolites to assess the intestinal metabolism of nitrosamines. Neither N-nitrosodiethylamine nor the two unsymmetrical nitrosamines were metabolized to any significant extent.
Nitrosodiethylamine has known human metabolites that include N-Nitrosoethanamine.

Toxicity Summary
IDENTIFICATION AND USE: N-nitrosodiethylamine (NDEA) is a yellow oil. It is used as a gasoline and lubricant additive, antioxidant and as a stabilizer in plastics. HUMAN STUDIES: NDEA has been identified as tobacco carcinogen. Short-term exposure of a bronchial epithelial cell line to smoking-equivalent concentrations of tobacco carcinogens including NDEA altered the expression of key proliferation regulatory genes, EGFR, BCL-2, BCL2L1, BIRC5, TP53, and MKI67, similar to that reported in biopsy specimens of pulmonary epithelium described to be preneoplastic lesions. ANIMAL STUDIES: NDEA caused tumors in several species of experimental animals, at several different tissue sites, and by several different routes of exposure. It was carcinogenic in animals exposed perinatally and as adults, causing tumors mainly in the liver, respiratory tract, kidney, and upper digestive tract. Benign and malignant liver tumors occurred in mice, rats, hamsters, guinea pigs, rabbits, dogs, and pigs orally exposed to NDEA. Liver tumors also occurred in rats following inhalation exposure or rectal administration; in mice, rats, and hamsters following intraperitoneal injection; in hamsters, guinea pigs, gerbils, and hedgehogs following subcutaneous injection; in mice following prenatal exposure; in birds following intramuscular injection; and in fish and frogs exposed to NDEA in the tank water. In dogs, exposure to NDEA by stomach tube followed by subcutaneous injection caused cancer of the liver and nasal cavity. Tumors of the lung and upper respiratory tract occurred in mice, rats, hamsters, dogs, and pigs following oral administration of NDEA. Tumors of the kidney occurred in rats following oral, intravenous, or prenatal administration of NDEA. Oral administration also caused kidney tumors in pigs and tumors of the upper digestive tract in mice, rats, and hamsters. The mutagenicity of NDEA was evaluated in Salmonella tester strains TA98, TA100, TA1535, TA1537 and TA1538 (Ames Test), both in the presence and absence of added metabolic activation. NDEA did not cause a reproducible positive response in any of the bacterial tester strains, either with or without metabolic activation. In the presence of liver microsomal fraction from phenobarbital-treated rats, NDEA caused 8-azaguanine-resistant mutants in Chinese hamster V79 cells. NDEA was mutagenic in recessive lethal test in Drosophila melanogaster. The effects of NDEA on sexual development, gametogenesis, and oocyte maturation were studied in Japanese medaka (Oryzias latipes). NDEA reduced the germ cell number dose-dependently during early stages of sexual differentiation in XX larvae, resulting in underdeveloped ovaries in adulthood at low doses. This effect was sex-specific as no such changes were seen in XY larvae. Furthermore, XX and XY larvae that were exposed at a low dose during early life showed a significant reduction in body weight in adulthood. Gonads in sexually immature adult medaka males and females exposed to NDEA were in advanced stages in comparison to that of the controls. ECOTOXICITY STUDIES: NDEA induced oxidative stress and antioxidant defense to zebrafish metabolism system at concentrations over 5 ug/L. After a 42-day exposure, a significant DNA damage was observed in zebrafish liver cells at NDEA concentrations beyond 500 ug/L. Toxic and carcinogenic effects observed in snakes (Phython reticulatus) after lifelong administration of 6, 12, and 24 mg/kg NDEA by gavage at four nightly intervals. Total dose needed to induce tumors was 500-600 mg/kg.

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Interactions
OBJECTIVE: The present paper aims to investigate the effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and N-nitrosodiethylamine (DEN) on tumorigenesis and its potential mechanism. METHODS: The potentials of TCDD and DEN in separation or in combination to induce malignant transformation were tested in Balb/c 3T3 cells by using a cell transformation assay method. The possible mechanism of observed effects was studied further by adding a-naphthoflavone (a-NF), a competitive binding agent of TCDD, to the Aryl hydrocarbon receptor (AhR) pathway. The mRNA expressions of Cyp1a1 and Cyp2a5 gene in Balb/c 3T3 cells treated by DEN and TCDD in separation or in combination with or without presence of a-NF were measured with fluorescence quantification RT-PCR technique. RESULTS: The cell transformation frequency (TF) was significantly higher in case of induction with TCDD in combination with DEN, as compared to that with either TCDD or DEN alone. These effects were not inhibited via a-NF. The mRNA expression levels of both Cyp1a1 and Cyp2a5 were enhanced by TCDD treatment alone, but this inducible effect was blocked in cells treated by TCDD and DEN in combination. CONCLUSION: TCDD and DEN had a significant synergistic effect on tumorigenesis when they were used in combination. AhR pathway may not be the key mechanism of this synergistic effect. Thus, it is necessary to further test the potential mechanism involved in cancer development.
... In the present study, we examined the effects of pitavastatin - a drug used for the treatment of hyperlipidemia - on the development of diethylnitrosamine (DEN)-induced liver preneoplastic lesions in C57BL/KsJ-db/db (db/db) obese mice. Male db/db mice were administered tap water containing 40 ppm DEN for 2 weeks and were subsequently fed a diet containing 1 ppm or 10 ppm pitavastatin for 14 weeks. At sacrifice, feeding with 10 ppm pitavastatin significantly inhibited the development of hepatic premalignant lesions, foci of cellular alteration, as compared to that in the untreated group by inducing apoptosis, but inhibiting cell proliferation. Pitavastatin improved liver steatosis and activated the AMPK-alpha protein in the liver. It also decreased free fatty acid and aminotransferases levels, while increasing adiponectin levels in the serum. The serum levels of tumor necrosis factor (TNF)-alpha and the expression of TNF-alpha and interleukin-6 mRNAs in the liver were decreased by pitavastatin treatment, suggesting attenuation of the chronic inflammation induced by excess fat deposition.
Resveratrol, a phytochemical compound abundant in red wine and grapes, is known to affect cancer cells both in vitro and in vivo. ... This study reports the effects of resveratrol in the early and advanced stages of hepatocarcinogenesis in a model of N-nitrosodiethylamine (DEN)-induced hepatocellular carcinoma (HCC) of male Wistar rats. For the experiment, rats were divided into different groups and treated with resveratrol either from day 1 of DEN administration for 15 days (pre-HCC), or after the development of HCC, i.e., 15-16 weeks after DEN administration (post-HCC), and compared to untreated HCC-bearing rats. Biochemical analysis of alpha-fetoprotein, the known serum marker for HCC, and other serum and liver marker enzymes also demonstrated a decreased level upon resveratrol treatment compared to the untreated HCC-bearing rats. H and E staining of tissue sections from the liver showed alteration or transformation of liver parenchymatous tissue in DEN-induced HCC (at 15-16 weeks). Resveratrol treatment during early (on day 1 of DEN-induction) and advanced (weeks 17-18) HCC showed a marked difference in the tissue architecture compared to untreated HCC. Immunoblot analysis revealed that resveratrol intervention at both the early and advanced stages of DEN-induced HCC activated the apoptotic markers, such as PARP cleavage, caspase-3 activation, p53 up-regulation and cytochrome-c release. In addition, semiquantitative RT-PCR and immunoblot analysis demonstrated the up- and down-regulation of key apoptotic regulators, such as Bax and Bcl2, respectively, in a resveratrol treatment-dependent manner. ...
The chemopreventive potential of Tephrosia purpurea extract (TPE) on N-nitrosodiethylamine (NDEA)-induced hepatocellular carcinoma (HCC) in Wistar rats was assessed. HCC was induced by a single intraperitoneal injection of NDEA (200 mg/kg) followed by subcutaneous injections of CCl(4) (3 mL/kg per week) for six weeks. After administration of the carcinogen, 200 and 400 mg/kg TPE were administered orally once a day throughout the study. The levels of liver cancer markers, including alpha-fetoprotein and carcinoembryonic antigen, were substantially increased by NDEA treatment. TPE treatment significantly reduced liver injury and restored the entire liver cancer markers. Additionally, TPE markedly normalized the activity of antioxidant enzymes, namely lipid peroxidation, reduced glutathione, catalase, superoxide dismutase, glutathione peroxidase and glutathione-S-transferase in the liver of NDEA-treated rats. Treatment with TPE significantly reduced the nodule incidence and multiplicity in the carcinogen-bearing rats. Histological observations of the liver tissues correlated with the biochemical observations. These findings powerfully support that T. purpurea prevented lipid peroxidation, suppressed the tumor burden, and promoted enzymatic and nonenzymatic antioxidant defence systems during NDEA-induced hepatocarcinogenesis. This might have been due to modulating the antioxidant defense status, which contributed to its anticarcinogenic potential.
For more Interactions (Complete) data for N-Nitrosodiethylamine (16 total), please visit the HSDB record page.

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Non-Human Toxicity Values
LD50 Rat subcutaneous 195 mg/kg
LD50 Rat intraperitoneal 216 mg/kg
LD50 Rat intravenous 280 mg/kg
LD50 Rat oral 280 mg/kg
For more Non-Human Toxicity Values (Complete) data for N-Nitrosodiethylamine (6 total), please visit the HSDB record page.

[1]. Impact of Deuterium Substitution on the Pharmacokinetics of Pharmaceuticals. Ann Pharmacother. 2019 Feb;53(2):211-216.C

n-Nitrosodiethylamine can cause cancer according to an independent committee of scientific and health experts.
N-nitrosodiethylamine is a clear slightly yellow liquid. Boiling point 175-177 °C. Can reasonably be anticipated to be a carcinogen. Used as a gasoline and lubricant additive and as an antioxidant and stabilizer in plastics.
N-nitrosodiethylamine is a nitrosamine that is N-ethylethanamine substituted by a nitroso group at the N-atom. It has a role as a mutagen, a hepatotoxic agent and a carcinogenic agent.
N-Nitrosodiethylamine is a synthetic light-sensitive, volatile, clear yellow oil that is soluble in water, lipids, and other organic solvents. It is used as gasoline and lubricant additive, antioxidant, and stabilizer for industry materials. When heated to decomposition, N-nitrosodiethylamine emits toxic fumes of nitrogen oxides. N-Nitrosodiethylamine affects DNA integrity, probably by alkylation, and is used in experimental research to induce liver tumorigenesis. It is considered to be reasonably anticipated to be a human carcinogen. (NCI05)
A nitrosamine derivative with alkylating, carcinogenic, and mutagenic properties.

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Mechanism of Action
... It is shown that the two nitrosamines N-nitrosodiethylamine and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone bind to nicotinic cholinergic receptors in hamster lung. Binding of the nitrosamines as well as nicotine to this receptor stimulates proliferation of human lung carcinoid cells in vitro. These data suggest chronic stimulation of nicotinic receptors by nicotine and nitrosamines in smokers as one of the molecular events responsible for stimulation of neuroendocrine cell proliferation and ultimately the development of lung tumors with neuroendocrine differentiation. ...

C4H6D4N2O

106.16

106.104

1346603-41-5

N-Nitrosodiethylamine;55-18-5

5921

Yellow oil
Slightly yellow liquid

0.9±0.1 g/cm3

173.9±9.0 °C at 760 mmHg

59.0±18.7 °C

1.7±0.3 mmHg at 25°C

1.442

0.42

0

3

2

7

51.7

0

CCN(CC)N=O

WBNQDOYYEUMPFS-VEPVEJTGSA-N

InChI=1S/C4H10N2O/c1-3-6(4-2)5-7/h3-4H2,1-2H3/i1D2,3D2

N-ethyl-N-(1,1,2,2-tetradeuterioethyl)nitrous amide

DEN-d4; Diethylnitrosamine-d4; N-Nitrosodiethylamine-d4

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

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

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.

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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).

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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)

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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).

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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

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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.

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 (Please use freshly prepared in vivo formulations for optimal results.)