In vitro, fibroblasts treated with trimethylamine N-oxide (TMAO) exhibited increased migration and size as compared to untreated fibroblasts. Trimethylamine N-oxide has the ability to upregulate the expression of collagen I and α-SMA while also increasing the expression of TGF-β receptor I, which in turn promotes the phosphorylation of Smad2. After treating newborn mouse fibroblasts with trimethylamine N-oxide, there is a decrease in the ubiquitination of TGF-βRI. Smurf2 expression is likewise inhibited by trimethylamine N-oxide [2]. Many marine animals have tissues that contain trimethylamine N-oxide, which offers protection from the damaging effects of hydrostatic pressure, high urea, temperature, and salt [3].

In animal modeling, cardiac fibrosis models can be created using trimethylamine N-oxide. Phenylacetate mustard (ip; 0–20 mg/kg; 15 days) has an ED15 value of 8.0 mg/kg and is consistently 1.8–1.9 times more potent against cancer than CHL [2]. 15% mortality is caused by 15.9 mg/kg of phenylacetic acid mustard (intraperitoneal injection; 0–20 mg/kg; single dosage) [2].

Metabolism / Metabolites
Trimethylamine N-oxide (TMAO) is biosynthesized in the liver from trimethylamine (TMA), which is derived from choline. Flavin monooxygenase 3 (FMO3) is involved in the oxidation of TMA; individuals with mutations in the FMO3 gene may experience an accumulation of TMA levels, leading to fishy odor syndrome. TMAO is excreted in the urine and is not further metabolized.

Toxicity Summary
Uremic toxins, such as trimethylamine oxide (TMAO), can be actively transported to the kidneys via organic ion transporters, particularly OAT3. Elevated uremic toxin levels can stimulate the production of reactive oxygen species (ROS). This appears to be mediated by the direct binding of uremic toxins to or inhibition of NADPH oxidases, particularly NOX4, which is abundant in the kidneys and heart (5). ROS can induce a variety of DNA methyltransferases (DNMTs) involved in the silencing of the KLOTHO protein. KLOTHO has been shown to play an important role in anti-aging, mineral metabolism, and vitamin D metabolism. Multiple studies have shown that KLOTHO mRNA and protein levels are reduced in acute or chronic kidney disease due to elevated local ROS levels (6). TMAO appears to contribute in part to the development of atherosclerosis by promoting the accumulation of cholesterol within macrophages, possibly through the induction of scavenger receptors such as CD36 and SRA1, both of which are involved in the uptake of modified lipoproteins (A15344).

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Toxicity Data
>A blood concentration of 100 μM usually indicates uremia.

[1]. High-choline Diet Exacerbates Cardiac Dysfunction, Fibrosis, and Inflammation in a Mouse Model of Heart Failure With Preserved Ejection Fraction. J Card Fail. 2020 May 14;S1071-9164(19)31802-0.

[2]. Gut Microbe-Derived Metabolite Trimethylamine N-oxide Accelerates Fibroblast-Myofibroblast Differentiation and Induces Cardiac Fibrosis. J Mol Cell Cardiol. 2019 Sep;134:119-130.

[3]. Trimethylamine N-Oxide: The Good, the Bad and the Unknown. Toxins (Basel). 2016 Nov 8;8(11):326.

Trimethylamine N-oxide (TMAO) is a tertiary amine oxide formed by the oxidation of the amino group of trimethylamine. It functions as an osmotic regulator, metabolite, and E. coli metabolite. Its function is related to trimethylamine. It is the conjugate base of hydroxytrimethylamine. TMAO is a metabolite found or produced in E. coli (K12 strain, MG1655 strain). TMAO has also been reported in grapes, Euglena, and other organisms with relevant data. TMAO is a uremic toxin, osmotic regulator, and atherosclerotic toxin (which can lead to atherosclerotic plaques). Uremic toxins can be classified into three main categories based on their chemical and physical properties: 1) small, water-soluble, non-protein-bound compounds, such as urea; 2) small, lipid-soluble and/or protein-bound compounds, such as phenols; and 3) larger, so-called medium-molecular-weight compounds, such as β2-microglobulin. Long-term exposure to uremic toxins can lead to various diseases, including kidney damage, chronic kidney disease, and cardiovascular disease. Trimethylamine N-oxide (TMAO) is an oxidation product of trimethylamine and a common metabolite in animals and humans. Specifically, TMAO is endogenously biosynthesized, while trimethylamine originates from choline, which in turn comes from dietary lecithin (phosphatidylcholine) or dietary carnitine. TMAO breaks down into trimethylamine (TMA), the primary source of the odor in rotting seafood. TMAO is an osmotic regulator used by the body to counteract the effects of elevated urea levels (caused by kidney failure), and its high levels can serve as a biomarker for kidney disease. Fishy odor syndrome, or trimethylamineuria, is caused by a deficiency in the production of flavin monooxygenase 3 (FMO3), resulting in the incomplete breakdown of trimethylamine in choline-containing foods into trimethylamine oxide. Trimethylamine then accumulates and is released through sweat, urine, and respiration, emitting a strong fishy odor. If bacteria in the gut can convert these substances into trimethylamine oxide (TMAO), then consuming foods containing carnitine or lecithin (phosphatidylcholine) will increase the concentration of TMAO in the blood. High concentrations of carnitine are found in red meat, certain energy drinks, and some dietary supplements; lecithin is found in eggs and is commonly used as an ingredient in processed foods. Many types of seafood also contain high concentrations of TMAO. Certain normal gut bacteria in the human gut (such as Acinetobacter) can convert dietary carnitine and lecithin into TMAO. TMAO alters cholesterol metabolism in the gut, liver, and arterial walls. When TMAO is present, cholesterol metabolism is altered, leading to increased cholesterol deposition in peripheral cells such as arterial walls and decreased cholesterol clearance (1, 2, 3).

C₃H₉NO

75.11

75.068

1184-78-7

Trimethylamine N-oxide dihydrate;62637-93-8;Trimethylamine N-oxide-d9;1161070-49-0;Trimethylamine-N-oxide-13C3

1145

White to off-white solid powder

0.9301 (rough estimate)

133.8°C (rough estimate)

220-222ºC(lit.)

1.4698 (estimate)

-2.57

0

1

0

5

28.4

0

UYPYRKYUKCHHIB-UHFFFAOYSA-N

InChI=1S/C3H9NO/c1-4(2,3)5/h1-3H3

N,N-dimethylmethanamine oxide

Trimethylamine Noxide; Trimethylamine N oxide

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)

H2O : ~100 mg/mL (~1331.38 mM)
DMSO : ~100 mg/mL (~1331.38 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (33.28 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% 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 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (33.28 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 25.0 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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (33.28 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 100 mg/mL (1331.38 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.

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