In murine macrophage RAW 264.7 cells, N,N-Dimethylacetamide (DMA) significantly suppressed nitric oxide (NO) production induced by lipopolysaccharide (LPS) stimulation in a concentration-dependent manner. [1]
Immunocytochemical analysis showed that DMA (at concentrations of 0.01 and 1 µmol/L) inhibited the LPS-induced nuclear translocation of the NF-κB p65 subunit in RAW 264.7 cells. The inhibition was more pronounced at the higher concentration (1 µmol/L). [1]
MTT assays confirmed that DMA at concentrations ranging from 0.1 to 10 µmol/L did not significantly affect the viability of RAW 264.7 cells, either in the presence or absence of LPS stimulation, indicating the observed effects were not due to cytotoxicity. [1]

In a murine model of LPS-induced preterm birth, intraperitoneal administration of DMA (at doses of 0.2, 0.39, 0.78, 1.56, and 3.1 mg/kg) prevented preterm delivery in timed-pregnant C57BL/6 mice in a dose-dependent manner. The proportion of mice delivering prematurely was significantly reduced at all doses except the lowest (0.2 mg/kg), with the highest doses (1.56 and 3.1 mg/kg) completely preventing delivery. [1]
DMA treatment also dose-dependently rescued pups from LPS-triggered spontaneous abortion. The number of pups lost was significantly lower in all DMA-treated groups compared to the LPS-only control, with complete rescue at doses of 1.56 and 3.1 mg/kg. [1]
Histological analysis of placental tissue revealed that DMA (1.56 mg/kg) significantly attenuated the robust inflammatory cell infiltration (primarily polymorphonuclear neutrophils) into the placental labyrinth induced by LPS. The tissue appearance in DMA-treated mice was similar to that of sham controls. [1]
Western blot analysis of placental tissue lysates showed that DMA (1.56 mg/kg) significantly decreased the expression of pro-inflammatory cytokines IL-1β, TNF-α, and IL-6, and reduced levels of matrix metalloproteinase-8 (MMP-8) compared to LPS-only controls. Concurrently, DMA significantly increased the expression of the anti-inflammatory cytokine IL-10. [1]
Examination of 265 pups exposed to DMA in utero revealed no evidence of congenital anomalies (abnormal extremities, craniofacial development, or abdominal wall defects). [1]

To assess the effect of DMA on macrophage function, RAW 264.7 cells were seeded and treated with various concentrations of DMA. One hour later, cells were stimulated with LPS (0.1 µg/mL). After 24 hours, cell culture supernatants were collected, centrifuged, and nitric oxide production was determined by measuring nitrite concentration using a Griess reagent assay. Absorbance was read at 540 nm. [1]
To evaluate the effect of DMA on NF-κB nuclear translocation, RAW 264.7 cells were seeded, allowed to adhere, and then treated with or without LPS (100 ng/mL) in the presence or absence of DMA (0.01 or 1 µmol/L) for 2 hours. Cells were fixed, permeabilized, blocked, and incubated overnight with an anti-NF-κB p65 primary antibody at 4°C. After washing, cells were incubated with a fluorescent dye-conjugated secondary antibody. Nuclei were counterstained. Translocation was assessed via immunofluorescence microscopy. [1]
To test the effect of DMA on cell viability, RAW 264.7 cells were seeded, allowed to adhere, and then stimulated with LPS (1 µg/mL) in the presence of a series of DMA concentrations for 24 hours. MTT solution was added and incubated for 2 hours. The formed formazan crystals were solubilized, and absorbance was measured at 570 nm using a microplate reader. [1]

Timed-pregnant C57BL/6 mice at embryonic day 15.5 (E15.5) received an intraperitoneal (i.p.) injection of LPS (50 mg/kg dissolved in 500 µL PBS). Mice were then randomly assigned to control or treatment groups. At 0.5 hours before and 10 hours after LPS injection, mice received i.p. injections of either 0.5 mL PBS (control) or 0.1 mL of varying concentrations of DMA (corresponding to doses of 0.2, 0.39, 0.78, 1.56, and 3.1 mg/kg). A sham group received PBS instead of LPS, followed by PBS injections. Mice were observed for preterm delivery for 24 hours, after which they were autopsied to confirm pregnancy status, count retained pups, and evaluate for fetal anomalies. Placentas were harvested for histological and molecular analysis. [1]

Absorption, Distribution and Excretion
N,N-Dimethylacetamide/…is readily absorbed through the skin…. N,N-Dimethylacetamide is rapidly absorbed across biofilms. No correlation was detected between individual airborne exposure and urinary excretion of monomethylacetamide over a full work shift (5 days). Of the 8 workers studied, the majority (n = 6) excreted approximately 13% of the calculated inhaled dose of metabolites via urine. DMAC is readily absorbed by the body after oral, skin, or inhalation exposure. …After inhalation, approximately 2% of the dimethylacetamide dose is excreted in urine as MMAC (N-methylacetamide); after inhalation plus skin exposure, approximately 10% of the dose is excreted. Exposure to 10 ppm of DMAC results in complete excretion of MMAC within 30 hours. The highest concentrations of MMAC in urine were 45 and 100 ppm (mg/L) in the inhalation and skin exposure groups, respectively. For workers continuously exposed to DMAC vapors (inhalation and skin contact) at airborne DMAC concentrations of 6–22 ppm, approximately 13.5% of the estimated dose is excreted in urine as MMAC. Some workers may excrete approximately 30% of the dose as MMAC. For more complete data on the absorption, distribution, and excretion of N,N-dimethylacetamide (7 metabolites), please visit the HSDB record page. Metabolites/Metabolites Dimethylacetamide undergoes N-demethylation in rats, resulting in the presence of the corresponding monomethylamide in urine… Acetamide, a didemethylation product of dimethylacetamide, has also been found. Analysis of urine from humans following occupational exposure to dimethylacetamide showed the presence of N-methylacetamide. /Excerpt from a table/ Gas chromatography analysis of urine from rats subcutaneously injected with dimethylacetamide revealed the presence of N-methylacetamide and acetamide in the urine. Both metabolites were also found in a mixture of dimethylacetamide and rat liver homogenate. N-Methylacetamide was detected in the urine of volunteers who inhaled or absorbed dimethylacetamide vapors through their skin. Measurements of the excreted metabolite N-methylacetamide in individuals exposed to dimethylacetamide vapors (with or without wearing masks that allowed inhalation of dimethylacetamide-free air) showed that more dimethylacetamide was absorbed through the lungs than through the skin. …Only 2–10% of inhaled dimethylacetamide was recovered from urine as N-methylacetamide. Studies have shown that the major urinary metabolite of dimethylformamide is N-(hydroxymethyl)-N-methylformamide, not N-methylformamide, because methanol-formamide decomposes on a gas chromatography column (to form N-methylformamide) but is relatively stable in aqueous solution. Similarly, it is reasonable to infer that the N-methylacetamide found in urine after exposure to dimethylacetamide is actually a product of the chemical decomposition of N-(hydroxymethyl)-N-methylacetamide during analysis. N,N-Dimethylacetamide is first metabolized into a monomethyl derivative via demethylation, and then into the parent acetamide.

Toxicity Data
LC50 (rat) = 2,475 ppm/1 hour

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Interactions Daily injection of progesterone (5 mg/hamster/day for 4 days) blocked the antifertility effect of DMA (administered at implantation), and injection of luteinizing hormone complex (1 mg/hamster/day prolactin plus 5.1 IU/hamster/day pregnant mare serum gonadotropin for 4 days) had the same effect. Dimethylacetamide reduced the incidence and/or yield of total tumors, benign plaques, benign hyperkeratotic lesions, and advanced tumors. Treatment with retinyl acetate or croton oil followed by 7,12-dimethylbenzo(a)anthracene initiation. In a single experiment, pyrimidine methylamine isothiocyanate was administered to rats at a dose of 45 mg/kg, dissolved in 6% dimethylacetamide, 6% dimethyl sulfoxide, or 95% ethanol. Results showed that the teratogenic effect was cumulative in dimethyl sulfoxide or ethanol; however, in dimethylacetamide, the dose-response relationship exhibited a parabolic curve. PCC4azal embryonic cancer cells were cultured in 129 strain mice via subcutaneous transplantation. When the tumor was palpable, it was treated with a combination of retinoic acid and dimethylacetamide. In vitro experiments showed that this embryonic cancer cell line had extremely low spontaneous differentiation capacity. The tumor was highly sensitive to differentiation induced by retinoic acid and/or dimethylacetamide. Intratumoral injection of 20 μl of a solution containing 10 mg retinoic acid/ml dimethylacetamide 10 times daily almost completely induced tumor morphological differentiation, primarily into neuroepithelial and glandular derivatives. Control group tumors showed only slight spontaneous differentiation. Differentiation was associated with decreased tumor growth rate, decreased mitotic index, reduced necrosis area, and prolonged host survival. In 4 of the 18 cases, long-term survival of the host was due to complete differentiation of the malignant embryonal carcinoma into a benign teratoma. Retinoic acid:dimethylacetamide, at the same dose and administration regimen, also effectively induced differentiation when administered systemically (i.e., intraperitoneally or subcutaneously).

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Non-human toxicity values
Oral LD50 in rats: 5.4 ml/kg
Intraperitoneal LD50 in mice: 3240 mg (3.4 ml)/kg
Oral LD50 in male rats: 5,809 mg/kg
Oral LD50 in female rats: 4,390 mg/kg
For more complete non-human toxicity data for N,N-dimethylacetamide (16 in total), please visit the HSDB record page.

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In this study, the doses of DMA used (up to 3.1 mg/kg) did not result in congenital abnormalities in the exposed mouse pups. [1] The article points out that DMA is used clinically as an excipient at doses far higher than the effective dose shown in this study. However, the article also notes that long-term exposure to DMA can lead to liver damage in humans. [1] In vitro MTT assays showed that DMA at concentrations up to 10 µmol/L had no significant effect on RAW cells. Cell viability was 264.7% within 24 hours. [1]

[1]. N,N-dimethylacetamide regulates the proinflammatory response associated with endotoxin and prevents preterm birth. Am J Pathol. 2013 Aug;183(2):422-30.

[2]. N,N-dimethylacetamide targets neuroinflammation in Alzheimer's disease in in-vitro and ex-vivo models. Sci Rep. 2023 May 1;13(1):7077.

[3]. FDA-Approved Excipient N, N-Dimethylacetamide Attenuates Inflammatory Bowel Disease in In Vitro and In Vivo Models. Fortune J Health Sci. 2022;5:499-509.

According to California labor law, N,N-dimethylacetamide is carcinogenic. An independent committee of scientific and health experts noted that it may cause developmental toxicity and male reproductive toxicity. Dimethylacetamide is a clear, colorless liquid with a slight ammonia odor. Its density is similar to water. Its flash point is 145°F (63°C). Its vapor is heavier than air. It can be absorbed through the skin and cause poisoning. It may irritate the eyes and skin. N,N-Dimethylacetamide solution (40% or less) is a colorless liquid with a slight ammonia odor. (US Coast Guard, 1999) N,N-Dimethylacetamide belongs to the acetamide class of compounds, its structure consisting of two hydrogen atoms on the nitrogen atom of acetamide replaced by two methyl groups. It is a metabolite observed in cancer metabolism. It functions as a human metabolite. It belongs to the acetamide class of compounds and is also a monocarboxylic acid amide. It is functionally related to acetamide.
N,N-Dimethylacetamide is a dipolar aprotic solvent and reagent that can be used in the production of fibers and pharmaceuticals, and also as a non-aldehyde fixative.
Hallucinogens are any natural or synthetic substances that can induce hallucinations.

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Therapeutic Uses
This commercial solvent has been tested as a parenteral drug delivery carrier and an antitumor agent.

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N,N-Dimethylacetamide (DMA) is described as an inexpensive, common aprotic organic solvent. This study identified it as a novel anti-inflammatory agent that can inhibit pro-inflammatory responses associated with endotoxin (LPS). [1]
The proposed mechanisms include DMA inhibiting macrophage activation, inhibiting NF-κB nuclear translocation, downregulating pro-inflammatory cytokines (IL-1β, TNF-α, IL-6), upregulating the anti-inflammatory cytokine IL-10, and reducing inflammatory cell infiltration, thereby preventing inflammation-mediated preterm birth. [1]
The authors believe that DMA is a potential non-toxic therapy for treating a variety of inflammatory diseases, but its in vivo and clinical validation for these indications still needs further evaluation. [1]

C4H9NO

87.1204

87.068

127-19-5

N,N-Dimethylacetamide-d9;16727-10-9;N,N-Dimethylacetamide-d6;31591-08-9;N,N-Dimethylacetamide-d3;20255-66-7

31374

Colorless to light yellow liquid

0.9±0.1 g/cm3

166.1±0.0 °C at 760 mmHg

-20 °C

70.0±0.0 °C

1.8±0.3 mmHg at 25°C

1.407

-0.75

0

1

0

6

58.6

0

O=C(C([H])([H])[H])N(C([H])([H])[H])C([H])([H])[H]

FXHOOIRPVKKKFG-UHFFFAOYSA-N

InChI=1S/C4H9NO/c1-4(6)5(2)3/h1-3H3

N,N-dimethylacetamide

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)

DMSO : ≥ 100 mg/mL (~1147.84 mM)

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.)