Endogenous Metabolite

ω-3 fatty acids reduce triglyceride (TG) levels, but corresponding increases in low-density lipoprotein cholesterol (LDL-C) levels may compromise achievement of lipid goals in patients with elevated cardiovascular risk. AMR101 is an investigational agent containing ≥96% of pure icosapent ethyl (the ethyl ester of eicosapentaenoic acid). The Phase III Multi-Center, Placebo-Controlled, Randomized, Double-Blind, 12-Week Study with an Open-Label Extension (MARINE) investigated the efficacy and safety of AMR101 in 229 patients with very high TG levels (≥500 mg/dl). AMR101 4 g/day significantly reduced median placebo-adjusted TG levels from baseline by 33.1% (p < 0.0001), and AMR101 2 g/day reduced TG levels by 19.7% (p = 0.0051). Changes in LDL-C were minimal and nonsignificant. AMR101 may offer substantial TG lowering without increases in LDL-C levels. [1]

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Thyroid hormones affect reactions in almost all pathways of lipid metabolism. It has been reported that plasma free fatty acid (FFA) concentration in hypothyroidism is generally within the normal range. In this study, however, we show that plasma FFA concentration in some hypothyroid patients is higher than the normal range. Symptoms of thyroid dysfunction in these individuals were less severe than those of patients with lower plasma FFA concentrations. From these findings we hypothesized that the change in FFA concentration must correlate with thyroid function. Using an animal model, we then examined the effect of highly purified eicosapentaenoic acid ethyl ester (EPA-E), a n-3 polyunsaturated fatty acid derived from fish oil, on thyroid function in 1-methyl-2-imidazolethiol (MMI)-induced hypothyroid rats. Oral administration of EPA-E inhibited reduction of thyroid hormone levels and the change of thyroid follicles in MMI-induced hypothyroid rats. These findings suggest that FFA may affect thyroid functions and EPA-E may prevent MMI-induced hypothyroidism[2].

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The precise mechanisms by which omega-3 fatty acids improve fat metabolism are not completely understood. This study was designed to determine the effects of eicosapentaenoic acid (EPA) ethyl ester administration on the expression levels of several muscle, liver and adipose tissue genes involved in lipogenesis and fatty acid oxidation pathways. Male Wistar rats fed a standard diet (control animals) or a high-fat diet were treated daily by oral gavage with EPA ethyl ester (1g/kg) for 5 weeks. The high-fat diet caused a very significant increase in plasma cholesterol (P<.01) levels, which was reverted by EPA (P<.001). A significant decrease in circulating triglyceride levels (P<.05) was also observed in EPA-treated groups. EPA administration induced a significant down-regulation in some lipogenic genes such as muscle acetyl CoA carboxylase beta (ACC beta) (P<.05) and liver fatty acid synthase (FAS) (P<.05). Furthermore, a decrease in glucokinase (GK) gene expression was observed in EPA-treated animals fed a control diet (P<.01), whereas a significant increase in GK mRNA levels was found in groups fed a high-fat diet. On the other hand, no alterations in genes involved in beta-oxidation, such acetyl CoA synthase 4 (ACS4), acetyl CoA synthase 5 (ACS5) or acetyl CoA oxidase (ACO), were found in EPA-treated groups. Surprisingly and opposite to the expectations, a very significant decrease in the expression levels of liver PPARalpha (P<.01) was observed after EPA treatment. These findings show the ability of EPA ethyl ester treatment to down-regulate some genes involved in fatty acid synthesis without affecting the transcriptional activation of beta-oxidation-related genes[3].

Twenty-nine male Wistar rats (6 weeks old) were housed in a temperature-controlled room (22±2°C) with a 12-h light–dark cycle. Animals were distributed into four experimental groups: control, control+EPA (CEPA), overweight and overweight+EPA (OEPA). All animals were maintained for an adaptation period of 4 days, fed chow diet and given deionized water ad libitum. After this period of time, the control and CEPA groups were fed a standard pelleted diet containing 76% carbohydrates, 6% lipids and 18% proteins (362 kcal/100 g). On the other hand, the overweight and OEPA groups were fed a cafeteria diet composed of the following items: paté, chips, bacon, chocolate, biscuits and pelleted diet (relative ratio, 2:1:1:1:1:1). The composition of this diet was as follows: 9% energy as protein, 29% energy as carbohydrate and 62% energy as lipid, by dry weight. All animals had ad libitum access to water and food for 5 weeks. The fatty acid composition of both control and high-fat diets was analyzed as previously reported, with the finding that both control and cafeteria diets have no EPA. Thus, the rats' only source of this fatty acid is oral gavage. Thus, the CEPA and OEPA groups were treated, simultaneously with the maintenance of diets for 35 days, with 1g/kg animal weight of highly purified EPA ethyl ester. This dose of EPA ethyl ester has been previously reported by Nobukata et al. to have a beneficial effect on diabetes prevention. The same volume of water was orally administrated to the control and overweight groups, as previously described in other studies, for 35 days. These control and overweight groups without treatment with any other type of fatty acid (such as saturated fatty acids with equal chain length as EPA or oleic acid) are more likely to be considered as control groups in our study design. This is because it has been demonstrated that supplementation with some other fatty acids is able to modify adiposity and circulating levels of biochemical and hormonal markers planned to be determined in the present study. [3]

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Five-week-old male Wistar rats were used in the exper- iments described in this study. The rats were divided into two groups of six on the basis of their initial body weights. Hypothyroidism in rats was induced by sub- cutaneous injection of MMI at a daily dose of 1 mg with con- comitant oral administration of Eicosapentaenoic Acid Ethyl Ester/EPA-E at a daily dose of 300 mg/kg (or the same volume of saline as a control) for 4 weeks. After 4 weeks the rats were anesthetized with chloroform and blood samples and thyroid tissues were obtained. Part of the thyroid tissues were immediately frozen in liquid nitrogen and stored at 80 C until use. [2]

Absorption, Distribution and Excretion
Icosapent ethyl is de-esterfied, converted into active EPA, and then absorbed in the small intestine. It reaches peak plasma concentration in 5 hours post-oral administration. Very little (<1%) is left circulating in the plasma as EPA incorporates into phospholipids, TG's, and cholesteryl esters.
Icosapent ethyl is not renally excreted
Steady state volume of distribution of active EPA is 88 L
Total plasma clearance, EPA = 684 mL/hr

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Metabolism / Metabolites
Once converted into active EPA, it is hepatically metabolized into acetyl Coenzyme A via beta-oxidation.

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Biological Half-Life
The half life of EPA is 89 hours.

9831415 rat LD50 oral >20 gm/kg Yakkyoku. Pharmacy., 41(1621), 1990
9831415 rat LD50 intraperitoneal 15 gm/kg Yakkyoku. Pharmacy., 41(1621), 1990
9831415 rat LD50 subcutaneous >20 gm/kg Yakkyoku. Pharmacy., 41(1621), 1990
9831415 mouse LD50 oral >20 gm/kg Yakkyoku. Pharmacy., 41(1621), 1990
9831415 mouse LD50 intraperitoneal >20 gm/kg Yakkyoku. Pharmacy., 41(1621), 1990

[1]. Jacobson TA. A new pure ω-3 eicosapentaenoic acid ethyl ester (AMR101) for the management of hypertriglyceridemia: the MARINE trial. Expert Rev Cardiovasc Ther. 2012 Jun;10(6):687-695.

[2]. Effect of eicosapentaenoic acid ethyl ester on hypothyroid function. J Endocrinol. 2001 Nov;171(2):259-65.

[3]. Down-regulation in muscle and liver lipogenic genes: EPA ethyl ester treatment in lean and overweight (high-fat-fed) rats. J Nutr Biochem. 2009 Sep;20(9):705-14.

Ethyl (5Z,8Z,11Z,14Z,17Z)-icosapentaenoate is a long-chain fatty acid ethyl ester resulting from the formal condensation of the carboxy group of (5Z,8Z,11Z,14Z,17Z)-icosapentaenoic acid with the hydroxy group of ethanol. It has a role as an anticholesteremic drug, a marine metabolite, an antipsychotic agent, an antidepressant and a prodrug. It is a long-chain fatty acid ethyl ester and a polyunsaturated fatty ester. It is functionally related to an all-cis-5,8,11,14,17-icosapentaenoic acid.
Icosapent ethyl or ethyl eicosapentaenoic acid is a synthetic derivative of the omega-3 fatty acid eicosapentaenoic acid (EPA). It is used as an adjunct therapy for severe hypertriglyceridemia (TG levels > 500 mg/dL) and to reduce the risk of cardiovascular events in certain patients with elevated triglycerides. FDA approved on July 26, 2012.
Icosapent Ethyl is a highly purified omega-3 fatty acid that can decrease serum triglyceride levels. Icosapent ethyl reduces serum triglycerides without an increase in LDL cholesterol, but increases the cholesterol and triglyceride content in skeletal muscle.
See also: Icosapent (has active moiety).

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Drug Indication
Icosapent ethyl is indicated as an adjunct to maximally tolerated statin therapy to reduce the risk of myocardial infarction, stroke, coronary revascularization, and unstable angina requiring hospitalizing in adult patients with elevated triglycerides (≥150 mg/dL) and established cardiovascular disease or who have diabetes mellitus and ≥2 other risk factors for cardiovascular disease. It is also indicated as an adjunct to diet to reduce triglyceride levels in adult patients with severe (≥500 mg/dL) hypertriglyceridemia.
FDA Label
Indicated to reduce cardiovascular risk as an adjunct to statin therapy.
Treatment of hypertriglyceridaemia

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Mechanism of Action
Studies suggest that EPA reduces hepatic very low-density lipoprotein triglycerides (VLDL-TG) synthesis and/or secretion and enhances TG clearance from circulating VLDL particles. Potential mechanisms of action include increased β-oxidation; inhibition of acyl-CoA:1,2-diacylglycerol acyltransferase (DGAT); decreased lipogenesis in the liver; and increased plasma lipoprotein lipase activity.

C22H34O2

330.512

330.255

C, 79.95; H, 10.37; O, 9.68

86227-47-6

Eicosapentaenoic Acid;10417-94-4;Eicosapentaenoic acid ethyl ester-d5;1392217-44-5

9831415

Colorless to light yellow liquid

0.9±0.1 g/cm3

417.0±34.0 °C at 760 mmHg

103.1±24.0 °C

0.0±1.0 mmHg at 25°C

1.496

7.32

0

2

15

24

425

0

C(/CCCC(=O)OCC)=C/C/C=C\C/C=C\C/C=C\C/C=C\CC

SSQPWTVBQMWLSZ-AAQCHOMXSA-N

InChI=1S/C22H34O2/c1-3-5-6-7-8-9-10-11-12-13-14-15-16-17-18-19-20-21-22(23)24-4-2/h5-6,8-9,11-12,14-15,17-18H,3-4,7,10,13,16,19-21H2,1-2H3/b6-5-,9-8-,12-11-,15-14-,18-17-

ethyl (5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenoate

Eicosapentaenoic Acid Ethyl Ester; Epadel; Miraxion; AMR-101; EPA-E; LAX-101; ethyl icosapentate; 86227-47-6; ICOSAPENT ETHYL; Eicosapentaenoic acid ethyl ester; Epadel; ethyl eicosapentaenoate; Vascepa; Timnodonic acid ethyl ester; EPA E; LAX 101; MND 21; MND-21; AMR 101; AMR101; MND21; EPAE; LAX101

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.

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 (~302.57 mM)
Ethanol : ~50 mg/mL (~151.29 mM)

Solubility in Formulation 1: ≥ 5 mg/mL (15.13 mM) (saturation unknown) in 10% EtOH + 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 50.0 mg/mL clear EtOH 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: 5 mg/mL (15.13 mM) (saturation unknown) in 10% EtOH + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 mg/mL clear EtOH 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: ≥ 5 mg/mL (15.13 mM) (saturation unknown) in 10% EtOH + 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 50.0 mg/mL clear EtOH stock solution to 900 μL of corn oil and mix well.

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Solubility in Formulation 4: 2.5 mg/mL (7.56 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
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 5: 2.5 mg/mL (7.56 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
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 6: ≥ 2.5 mg/mL (7.56 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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 (Please use freshly prepared in vivo formulations for optimal results.)