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 is biosynthesized in the liver from trimethylamine (TMA), which is derived from choline. Flavin monooxygenase 3 (FMO3) has been implicated in the oxidation of TMA since individuals with mutations in FMO3 present with accumulation of TMA levels, causing fish malodor syndrome. TMAO is secreted in the urine and is not metabolized any further.

Toxicity Summary
Uremic toxins such as TMAO are actively transported into the kidneys via organic ion transporters (especially OAT3). Increased levels of uremic toxins can stimulate the production of reactive oxygen species. This seems to be mediated by the direct binding or inhibition by uremic toxins of the enzyme NADPH oxidase (especially NOX4 which is abundant in the kidneys and heart) (5). Reactive oxygen species can induce several different DNA methyltransferases (DNMTs) which are involved in the silencing of a protein known as KLOTHO. KLOTHO has been identified as having important roles in anti-aging, mineral metabolism, and vitamin D metabolism. A number of studies have indicated that KLOTHO mRNA and protein levels are reduced during acute or chronic kidney diseases in response to high local levels of reactive oxygen species (6). TMAO appears to contribute to the development of atherosclerosis in part by promoting cholesterol accumulation within macrophages, perhaps by inducing 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
>100 uM in blood is usually indicative of 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 is a tertiary amine oxide resulting from the oxidation of the amino group of trimethylamine. It has a role as an osmolyte, a metabolite and an Escherichia coli metabolite. It is functionally related to a trimethylamine. It is a conjugate base of a hydroxytrimethylaminium.
Trimethylamine N-Oxide is a metabolite found in or produced by Escherichia coli (strain K12, MG1655).
Trimethylamine oxide has been reported in Vitis vinifera, Euglena gracilis, and other organisms with data available.
TMAO is a uremic toxin, an osmolyte and an atherotoxin (causing atherosclerotic plaques). Uremic toxins can be subdivided into three major groups based upon their chemical and physical characteristics: 1) small, water-soluble, non-protein-bound compounds, such as urea; 2) small, lipid-soluble and/or protein-bound compounds, such as the phenols and 3) larger so-called middle-molecules, such as beta2-microglobulin. Chronic exposure of uremic toxins can lead to a number of conditions including renal 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. In particular, trimethylamine-N-oxide is biosynthesized endogenously from trimethylamine, which is derived from choline, which can be derived from dietary lecithin (phosphatidylcholines) or dietary carnitine. TMAO decomposes to trimethylamine (TMA), which is the main odorant that is characteristic of degrading seafood. TMAO is an osmolyte that the body will use to counteract the effects of increased concentrations of urea (due to kidney failure) and high levels can be used as a biomarker for kidney problems. Fish odor syndrome or trimethylaminuria is a defect in the production of the enzyme flavin containing monooxygenase 3 (FMO3) causing incomplete breakdown of trimethylamine from choline-containing food into trimethylamine oxide. Trimethylamine then builds up and is released in the person's sweat, urine, and breath, giving off a strong fishy odor. The concentration of TMAO in the blood increases after consuming foods containing carnitine or lecithin (phosphatidylcholines), if the bacteria that convert those substances to TMAO are present in the gut. High concentrations of carnitine are found in red meat, some energy drinks, and certain dietary supplements; lecithin is found in eggs and is commonly used as an ingredient in processed food. High levels of TMAO are found in many seafoods. Some types of normal gut bacteria (e.g. species of Acinetobacter) in the human gut convert dietary carnitine and dietary lecithin to TMAO. TMAO alters cholesterol metabolism in the intestines, in the liver and in arterial wall. When TMAO is present, cholesterol metabolism is altered and there is an increased deposition of cholesterol within, and decreased removal of cholesterol from, peripheral cells such as those in the artery wall (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.)