Composition for reducing serum TMAO

Abstract

To develop a new use of fish oil components, particularly a new use thereof on health promotion.SOLUTION: A composition for reducing trimethylamine N-oxide in blood serum, containing a component selected from mono-unsaturated fatty acid having 20 or more carbon atoms, salts thereof and esters thereof as an active ingredient; the active ingredient is derived from fish oil; and the fish oil is derived from saury oil.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to compositions for reducing serum levels of trimethylamine N-oxide (TMAO). [Background technology]

[0002] It is known that fish are rich in n-3 polyunsaturated fatty acids, which are unsaturated fatty acids. Many studies have been reported to date that have shown that frequent consumption of fish reduces the risk of developing cardiovascular disease. In particular, an epidemiological study in Greenland is famous as a pioneering epidemiological study that drew attention to the n-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA) in the prevention and treatment of cardiovascular disease. In this epidemiological study, it was revealed that the indigenous Inuit people of Greenland had a low incidence of acute myocardial infarction and a low mortality rate from ischemic heart disease. Furthermore, an investigation of their diet revealed that they consumed a lot of n-3 polyunsaturated fatty acids such as EPA and docosahexaenoic acid (DHA) due to their dietary habit of eating a lot of seals and fish, and conversely, they consumed little n-6 polyunsaturated fatty acids, which are abundant in terrestrial animals and plants. It has been reported that such differences in the fatty acid composition ingested in the diet are the reason for the low incidence of acute myocardial infarction and the low mortality rate from ischemic heart disease in the Inuit (Non-Patent Documents 1 to 3). In addition, human clinical trials have revealed the neutral fat lowering effect of EPA and DHA, but it is known that the cholesterol lowering effect is almost nonexistent or weak.

[0003] On the other hand, it has been reported that long-chain monounsaturated fatty acids (LC-MUFA) with carbon numbers of 20 and 22, which are known to be abundant in saury oil and Alaska pollock oil, have a cholesterol-lowering effect, unlike EPA and DHA (Patent Document 1). In addition, in diet-induced obesity model mice and type 2 diabetes model mice, the effects of consuming saury oil and Alaska pollock oil, such as lowering total cholesterol, LDL-cholesterol, triglyceride, and blood glucose levels in the blood and improving insulin resistance, have been found (Non-Patent Documents 4 to 6). A similar effect has also been confirmed in oils and fats selectively enriched with LC-MUFA (Non-Patent Document 5). In relation to this, a cohort study of 50,000 Japanese men and women has reported that men who consume more seafood have a lower risk of developing diabetes, and that the risk is particularly lower in groups that consume a large amount of small and medium-sized fish such as saury and mackerel, and fatty fish such as salmon (Non-Patent Document 7).

[0004] Furthermore, a study using an arteriosclerosis model mouse reported that intake of LC-MUFA inhibited the formation of atherosclerotic lesions in the aortic sinus, and that this was likely the result of a combined protective effect of reducing blood inflammatory cytokine concentrations and promoting cholesterol removal from macrophages by stimulating the PPAR signaling pathway (Non-Patent Document 8).

[0005] The present inventors have also found that ingestion of oil rich in LC-MUFA improves flow-mediated vasodilation (Patent Document 2).

[0006] Trimethylamine N-oxide (TMAO) is a substance that is produced by metabolism of trimethylamine (TMA)-containing substances in foods, such as phosphatidylcholine, choline, and carnitine, through intestinal bacterial metabolism and metabolism in the liver. Associations between TMAO and the onset of various diseases, such as cardiovascular diseases such as atherosclerosis (Non-Patent Documents 9-11), heart failure (Non-Patent Document 12), thrombosis (Non-Patent Document 13), and chronic kidney disease (Non-Patent Document 14), have been reported. [Prior art documents] [Chartered documents]

[0007]

Patent Document 1

Patent document 2

Non-licensed literature

[0008]

Non-licensed literature 1

Non-licensed Document 4

Non-licensed Document 5

Non-licensed Document 6

Non-licensed Document 7

Non-licensed Document 8

Non-licensed literature 9

[0009] It has been reported that elevated serum levels of TMAO are associated with various diseases, and there is a need for a method to reduce serum levels of TMAO that is safe and can be implemented for the long term. In addition, among LC-MUFA, positional isomers such as C20:1n-11 and C22:1n-11 are also found in fish oils derived from fish species other than saury, such as Alaska pollock, mackerel, and salmon, but are hardly found in vegetable oils. Since saury and Alaska pollock are popular fish among Japanese people and have high industrial value, there is a strong demand for the development of new applications for fish oils rich in LC-MUFA prepared from these fish species.

[0010] The present invention aims to provide new uses of fish oil components containing LC-MUFA, particularly new uses relating to health promotion. [Means for solving the problem]

[0011] As a result of intensive research conducted by the present inventors to achieve the above-mentioned object, they discovered that the intake of saury oil, which is rich in LC-MUFA, reduces serum levels of TMAO. It was not previously known that oils rich in LC-MUFA affect serum levels of TMAO. Based on these findings, the present inventors conducted further research and completed the present invention. That is, the present invention is as follows.

[0012] [1] A composition for reducing serum trimethylamine N-oxide, comprising: The composition contains, as an active ingredient, a component selected from monounsaturated fatty acids having 20 or more carbon atoms, salts thereof, and esters thereof. [2] The composition according to [1], wherein the active ingredient is a component selected from a monounsaturated fatty acid having 20 carbon atoms, a salt thereof and an ester thereof, a component selected from a monounsaturated fatty acid having 22 carbon atoms, a salt thereof and an ester thereof, or a combination thereof. [3] The composition according to [1] or [2], wherein the active ingredient is a glyceride containing a monounsaturated fatty acid having 20 or more carbon atoms as a constituent fatty acid. [4] The composition described in [3], wherein the glyceride is a triglyceride. [5] The composition according to any one of [1] to [4], further comprising an n-3 polyunsaturated fatty acid and an n-6 polyunsaturated fatty acid, wherein the weight ratio of the n-3 polyunsaturated fatty acid to the n-6 polyunsaturated fatty acid is 4.0 or more. [6] The composition according to any one of [1] to [5], wherein the active ingredient is derived from fish oil. [7] The composition described in [6], wherein the fish oil is saury oil. [8] The composition according to any one of [1] to [7], characterized in that the composition is used so that 2 mg / kg body weight / day or more of the active ingredient calculated as a monounsaturated fatty acid having 20 or more carbon atoms is orally administered for 4 weeks or more. [9] The composition according to any one of [1] to [8], which is for reducing the risk of contracting a disease caused by an increase in serum level of trimethylamine N-oxide.

[10] The composition according to any one of [1] to [9], which is a food composition.

[11] The composition according to any one of [1] to [9], which is a pharmaceutical composition. Effect of the Invention

[0013] The present invention provides a composition for reducing serum TMAO that contains a fish oil component rich in LC-MUFA, such as saury oil, and can reduce the risk of contracting diseases caused by elevated serum levels of TMAO. [Brief description of the drawings]

[0014] [Figure 1] 1 is a graph showing the change in serum TMAO levels before and after ingestion of subjects who took the saury oil capsule or blended oil capsule of Example 1. The saury oil capsule group: p=4.3×10-9; the blended oil capsule group: p=1.0×10-4. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] The present invention will now be described in more detail. In the present specification, the following abbreviations may be used. TMAO: Trimethylamine N-oxide MUFA: Monounsaturated fatty acids LC-MUFA: A general term for long-chain monounsaturated fatty acids, especially the C20:1 and C22:1 isomers PUFA: Polyunsaturated fatty acids

[0016] <Composition for reducing serum TMAO> The present invention provides a composition for reducing serum TMAO, which contains, as an active ingredient, a component selected from monounsaturated fatty acids having 20 or more carbon atoms, their salts, and their esters (hereinafter, sometimes referred to as the composition of the present invention).

[0017] The monounsaturated fatty acid having 20 or more carbon atoms, its salt, or its ester used in the present invention is not particularly limited, and may be any one that is acceptable for pharmaceutical or food use. Glycerides containing monounsaturated fatty acids having 20 or more carbon atoms as constituent fatty acids can be produced by known methods, and can also be produced as oils and fats derived from natural products, for example, by the boiling method described in WO1996 / 26647. Examples of oils and fats derived from natural products include marine oils (for example, fish oils such as saury oil, and mammalian oils and fats such as seals and whales), microbial oils, etc. As described later, fish oils have a high content of monounsaturated fatty acids having 20 or more carbon atoms. Therefore, it is preferable that the active ingredient in the composition of the present invention is derived from fish oil (for example, saury oil). In addition, free monounsaturated fatty acids having 20 or more carbon atoms, their salts, and esters other than glycerides can be prepared by known methods, for example, using the above-mentioned glycerides as raw materials.

[0018] For example, saury crude oil is usually extracted in the same way as other fish oils: Whole saury or processing residues from seafood processing such as fish heads, skin, backbone, and innards are crushed and steamed, then pressed to separate the broth (stick water, SW) and pressed meal. The oil obtained with the broth is separated from the broth by centrifugation.

[0019] The 2015 edition (7th revision) of the Standard Tables of Food Composition in Japan states that the fatty acids in saury (skinned, raw) contain 26.0% docosenoic acid (C22:1) by weight, 17.6% eicosenoic acid (C20:1) by weight, and 54.2% total monounsaturated fatty acids. Among fish oils, saury oil is characterized by its high content of monounsaturated fatty acids.

[0020] Crude fish oil is refined into refined fish oil through refining processes such as degumming, deacidification, bleaching, deodorization, etc. This refined fish oil can also be used as a supply source of a component selected from monounsaturated fatty acids having 20 or more carbon atoms, their salts, and their esters.

[0021] In one embodiment, an oil having an increased concentration of monounsaturated fatty acids may be used as a supply source of a component selected from monounsaturated fatty acids having 20 or more carbon atoms, their salts, and their esters. In this case, a triglyceride enriched in MUFA can be obtained by a method of concentrating the monounsaturated fatty acid ethyl esters after ethyl esterification, and then transesterifying the monounsaturated fatty acid with glycerin to reconstitute the triglyceride.

[0022] In one embodiment, the composition of the present invention may be a fatty acid composition mainly composed of free fatty acid, a fatty acid salt composition mainly composed of fatty acid salt, or a fatty acid ester composition mainly composed of fatty acid ester. Examples of fatty acid salts include potassium salts and sodium salts. Examples of fatty acid esters include esters of lower alcohols having 5 or less carbon atoms (i.e., alkyl esters; for example, methyl esters, ethyl esters, n-propyl esters, i-propyl esters, n-butyl esters, s-butyl esters, t-butyl esters, n-pentyl esters, etc.), esters of glycerin such as monoglycerides, diglycerides, and triglycerides (i.e., glycerides), and phospholipids. In a preferred embodiment, the composition of the present invention is a fatty acid alkyl ester composition or a fatty acid glyceride composition, and more preferably a fatty acid ethyl ester composition or a fatty acid triglyceride composition.

[0023] Examples of monounsaturated fatty acids having 20 or more carbon atoms include monounsaturated fatty acids having 20 carbon atoms (C20:1) (e.g., C20:1 n-11, C20:1 n-9, C20:1 n-7), monounsaturated fatty acids having 22 carbon atoms (C22:1) (e.g., C22:1 n-11, C22:1 n-9, C22:1 n-7, C22:1 n-13), and monounsaturated fatty acids having 24 carbon atoms (C24:1) (e.g., C24:1 n-9). In a preferred embodiment, the active ingredient of the composition of the present invention is a component selected from C20:1, a salt thereof, and an ester thereof, a component selected from C22:1, a salt thereof, and an ester thereof, or a combination thereof.

[0024] The content of C22:1 and C20:1 varies depending on the fish species, but examples of fish species with high content of these include fish belonging to the family Sauryidae such as Pacific cod, Alaska pollock, Atlantic cod, and sablefish, fish belonging to the family Gadidae such as Chum salmon, Coho salmon, Sockeye salmon, Pink salmon, Pacific salmon, and Rainbow trout, fish belonging to the family Osmeridae such as Capelin and Shishamo, and fish belonging to the family Clupeidae such as Herring. In addition, fish such as sand lance, tuna, mackerel, and alfonsino are relatively rich in these. They are also rich in the liver oil of sharks such as the spiny dogfish, basking shark, and chimaera. In the present invention, fish oils prepared from these fish can be used as they are, purified, or concentrated.

[0025] The ratio of monounsaturated fatty acids having 20 or more carbon atoms to the total fatty acids in the composition of the present invention can be 10% by weight or more, for example, 11% by weight or more, 12% by weight or more, 13% by weight or more, 14% by weight or more, 15% by weight or more, 16% by weight or more, 17% by weight or more, 18% by weight or more, 19% by weight or more, 20% by weight or more, 21% by weight or more, 22% by weight or more, 23% by weight or more, 24% by weight or more, 25% by weight or more, 26% by weight or more, 27% by weight or more, 28% by weight or more, 29% by weight or more, 30% by weight or more, 40% by weight or more, 50% by weight or more, 60% by weight or more, 70% by weight or more, 80% by weight or more, or 90% by weight or more. In the present invention, the ratio of a specific fatty acid to the total fatty acids in the composition (or the content of a specific fatty acid to the total weight of fatty acids) is determined based on the fatty acid composition. The fatty acid composition can be determined according to a conventional method. Specifically, the oil to be measured is esterified using a lower alcohol and a catalyst to obtain a fatty acid lower alkyl ester. The obtained fatty acid lower alkyl ester is then analyzed using a gas chromatograph equipped with a flame ionization detector (FID). Peaks corresponding to each fatty acid are identified in the obtained gas chromatography chart, and the peak area of ​​each fatty acid is determined using the Agilent ChemStation integration algorithm (revision C.01.03

[37] , Agilent Technologies). The fatty acid composition is determined by the percentage of each peak area relative to the sum of the peak areas of the fatty acids. The area % obtained by the above-mentioned measurement method is the same as the weight % of each fatty acid in the sample. Please refer to the Japan Oil Chemists' Society (JOCS) Standard Testing Methods for Analysis of Fats, Oils and Lubricants, 2013 Edition, 2.4.2.1-2013 Fatty Acid Composition (FID Constant Temperature Gas Chromatography) and 2.4.2.2-2013 Fatty Acid Composition (FID Programmed Temperature Gas Chromatography). The analysis conditions for gas chromatography are as follows. Equipment: Agilent 6890N GC system (Agilent) Column: DB-WAX (Agilent Technologies, 30m x 0.25mm ID, 0.25μm film thickness) Carrier gas: Helium (1.0 mL / min, constant flow) Inlet temperature: 250℃ Injection volume: 1μL Injection method: Split Split ratio: 20:1 Column oven: 180℃ - 3℃ / min - 230℃ Detector: FID Detector temperature: 250℃

[0026] In this specification, the fatty acid content also means the fatty acid content (wt%) determined based on the above fatty acid composition.

[0027] In the present invention, examples of the salts of monounsaturated fatty acids having 20 or more carbon atoms include potassium salts and sodium salts. Examples of esters of monounsaturated fatty acids having 20 or more carbon atoms include esters of lower alcohols having 5 or less carbon atoms (e.g., methyl esters, ethyl esters, n-propyl esters, i-propyl esters, n-butyl esters, s-butyl esters, t-butyl esters, n-pentyl esters, etc.), esters with glycerin such as monoglycerides, diglycerides, and triglycerides (i.e., glycerides), and phospholipids. In a preferred embodiment, the active ingredient of the composition of the present invention is a glyceride containing a monounsaturated fatty acid having 20 or more carbon atoms as a constituent fatty acid, and more preferably a triglyceride.

[0028] As the monounsaturated fatty acid having 20 or more carbon atoms in the present invention, fats and oils derived from natural products containing free monounsaturated fatty acids having 20 or more carbon atoms may be used as they are, or after purification or concentration. As the ester of monounsaturated fatty acids having 20 or more carbon atoms, fats and oils derived from natural products containing glycerides having monounsaturated fatty acids having 20 or more carbon atoms as constituent fatty acids may be used as they are, or after purification or concentration.

[0029] Also, as the ester of monounsaturated fatty acids having 20 or more carbon atoms, fractionated oils obtained by esterifying fats and oils derived from natural products and then fractionating them by high performance liquid chromatography, etc., can be used. Examples of fractionated oils include fractionated oils enriched in C20:1, fractionated oils enriched in C22:1, and fractionated oils enriched in C24:1.

[0030] The composition of the present invention may further comprise n-3 polyunsaturated fatty acids and n-6 ​​polyunsaturated fatty acids, in which the weight ratio of n-3 polyunsaturated fatty acids to n-6 polyunsaturated fatty acids is 4.0 or more, for example 5.0 or more, 6.0 or more, 7.0 or more, 8.0 or more, 9.0 or more, or 10.0 or more. In this specification, n-3 polyunsaturated fatty acid refers to an unsaturated fatty acid having 18 or more carbon atoms and two or more carbon-carbon double bonds, in which the first double bond is between the third and fourth carbon atoms counting from the carbon of the methyl group at the end of the fatty acid chain. Examples of such fatty acids include α-linolenic acid (C18:3 n-3), octadecatetraenoic acid (C18:4 n-3), eicosatetraenoic acid (C20:4 n-3), eicosapentaenoic acid (C20:5 n-3), docosapentaenoic acid (C22:5 n-3), and docosahexaenoic acid (C22:6 n-3). The n-6 polyunsaturated fatty acid refers to an unsaturated fatty acid having 18 or more carbon atoms and two or more carbon-carbon double bonds, in which the first double bond is between the 6th and 7th carbon atoms counting from the carbon of the methyl group at the end of the fatty acid chain, such as linoleic acid (C18:2n-6) and arachidonic acid (C20:4n-6). In this specification, the weight ratio of fatty acids refers to the weight ratio based on the fatty acid composition determined as described above. The gas chromatography analysis conditions are as described above.

[0031] The composition of the present invention can reduce the serum level of TMAO in a subject.Therefore, in one embodiment, a method for reducing the serum level of TMAO in a subject is provided, comprising administering the composition of the present invention to the subject. In healthy individuals, the plasma level of TMAO is less than 2.5 μM in the fasting state. Prospective follow-up of individuals with high plasma TMAO levels has revealed that they have a higher incidence of cardiovascular events, and TMAO levels higher than 6.2 μM significantly increase the risk of major adverse cardiovascular events (analysis of approximately 4,000 subjects divided into quartile groups (<2.43, 2.43-3.66, 3.67-6.18, and >6.18) based on plasma TMAO concentration; see Non-Patent Document 10). The composition of the present invention can prevent the serum level of TMAO from increasing to a level that increases the risk of developing cardiovascular disease (e.g., 2.5 μM or more, 3.7 μM or more, or 6.2 μM or more). The compositions of the present invention can also reduce serum levels of TMAO to less than 2.5 μM, less than 2.4 μM, less than 2.3 μM, less than 2.2 μM, less than 2.1 μM, less than 2.0 μM, less than 1.9 μM, less than 1.8 μM, less than 1.7 μM, less than 1.6 μM, or less than 1.5 μM. The compositions of the present invention can also reduce serum levels of TMAO after ingestion by 0.10 μM or more, 0.15 μM or more, 0.20 μM or more, 0.25 μM or more, 0.29 μM or more, 0.50 μM or more, 1.0 μM or more, 1.5 μM or more, 2.0 μM or more, 2.5 μM or more, 3.0 μM or more, 3.5 μM or more, or 4.0 μM or more compared to serum levels of TMAO before ingestion. Known methods for measuring the serum level of TMAO include, for example, stable isotope dilution mass spectrometry (see Patent Document 9 above) and ELISA. In a preferred embodiment of the present invention, the serum level of TMAO is measured by ELISA. Measurement by ELISA can be performed by a standard method using an antibody against TMAO. Alternatively, a commercially available kit such as HUMAN Trimethylamine-N-oxide (TMAO) ELISA KIT (Glory Science CO., LTD) may be used.

[0032] The test in the examples described below is conducted on healthy subjects, and the reduction of serum levels of TMAO does not mean the treatment of diseases such as cardiovascular disease, but it can reduce the risk of suffering from such diseases. Thus, in one aspect, the composition of the present invention is a composition for reducing the risk of suffering from diseases caused by an increase in serum levels of TMAO. In another aspect, a method for reducing the risk of suffering from diseases caused by an increase in serum levels of TMAO is provided, comprising administering the composition of the present invention to a subject. Here, examples of diseases caused by an increase in serum levels of TMAO include cardiovascular diseases such as atherosclerosis, heart failure, and thrombosis, and chronic kidney disease. The present invention also provides a novel use of LC-MUFA that has not been known before, based on the discovery that LC-MUFA reduces serum levels of TMAO.Therefore, in one embodiment, the composition of the present invention can be a composition for reducing the risk of contracting a disease caused by an increase in serum levels of TMAO, excluding arteriosclerosis.In another embodiment, a method for reducing the risk of contracting a disease caused by an increase in serum levels of TMAO, excluding arteriosclerosis, is provided, comprising administering the composition of the present invention to a subject.

[0033] In one embodiment, the above-mentioned method for reducing the serum level of TMAO in a subject and the method for reducing the risk of suffering from a disease caused by an increase in the serum level of TMAO in a subject can be a non-therapeutic method, where "non-therapeutic" does not include medical procedures, i.e., surgery, treatment, or diagnosis performed by a doctor (including a person under the instruction of a doctor) on a human being.

[0034] The subject of administration of the composition of the present invention is a mammal, preferably a human. The age of the subject is not particularly limited as long as the effect of the present invention can be obtained, for example, 20 years or older. In one embodiment, the subject of administration may be a healthy person. In another embodiment, the subject of administration may be a mammal that does not suffer from a disease caused by an increase in serum level of TMAO. In yet another embodiment, the subject of administration may be a mammal that does not suffer from a disease caused by an increase in serum level of TMAO, except for arteriosclerosis.

[0035] The composition of the present invention may be prepared in a form suitable for pharmaceutical compositions, food compositions (e.g., functional foods, health foods, supplements, etc.), for example, in the form of various solid preparations such as granules (including dry syrups), capsules (soft capsules, hard capsules), tablets (including chewable tablets, etc.), powders (powdered agents), pills, etc., or liquid preparations such as oral liquids (including liquids, suspensions, syrups, etc.). For example, the composition of the present invention can be formulated as a soft capsule in which purified fish oil is filled in a gelatin membrane.

[0036] Examples of additives for formulation include excipients, lubricants, binders, disintegrants, fluidizers, dispersants, wetting agents, preservatives, thickeners, pH adjusters, colorants, flavorings, surfactants, and dissolution aids. When the formulation is in the form of a liquid, a thickener such as pectin, xanthan gum, or guar gum can be added. In addition, the formulation can be made into a coated tablet using a coating agent, or into a paste-like glue. Furthermore, when preparing other forms, conventional methods can be followed.

[0037] The composition of the present invention can take the form of a food composition. In the present invention, the food composition means food in general, including beverages, and includes general foods, including health foods such as supplements, as well as specific health foods and nutritional functional foods as specified by the Consumer Affairs Agency's Health Food Function Food System. For example, functional foods labeled with the effect of lowering serum levels of TMAO are provided. For example, foods containing fish oil can be provided as they are. In addition, the food composition of the present invention also includes food materials that can be added to, mixed with, or applied to other foods to impart the effect of lowering serum levels of TMAO. In addition to foods, the food composition can also be provided as animal feed, etc.

[0038] When the composition of the present invention is in the form of food, the food is not particularly limited and may be beverages, confectioneries, breads, soups, etc., and includes, for example, general retort foods, frozen foods, instant foods (such as noodles), canned foods, sausages, cookies, biscuits, cereal bars, crackers, snacks (such as potato chips), pastries, cakes, pies, candies, chewing gums (including pellets and sticks), jellies, soups, ice cream, dressings, yogurt, etc., supplements in the form of tablets, capsules, emulsions, etc., soft drinks, etc. The manufacturing method of these foods is not particularly limited as long as it does not impair the effects of the present invention, and may follow the method used by those skilled in the art for each food.

[0039] The present invention also includes the implementation of the present invention in which the effects of the composition of the present invention are displayed on packaging containers (boxes, bags, cans, bottles, etc.), product instructions, pamphlets, etc., and in which the product of the present invention is sold. The present invention also includes the implementation of the present invention in which the effects of the present invention are displayed on television, Internet websites, pamphlets, newspapers, magazines, etc., and in which the product of the present invention is advertised and sold.

[0040] In the present invention, the amount of the component selected from monounsaturated fatty acids having 20 or more carbon atoms, their salts, and their esters ingested by a subject is not particularly limited, and may be ingested in an amount equal to or greater than the effective amount for obtaining the desired effect, where the effective amount for obtaining the desired effect refers to the amount required to reduce the serum level of TMAO. For example, in the case of adults, depending on the age, weight, health condition, and other conditions of the subject, the daily intake may be 2 mg / kg body weight / day or more, e.g., 3 mg / kg body weight / day or more, 4 mg / kg body weight / day or more, 5 mg / kg body weight / day or more, 6 mg / kg body weight / day or more, 7 mg / kg body weight / day or more, 8 mg / kg body weight / day or more, 9 mg / kg body weight / day or more, 10 mg / kg body weight / day or more, 11 mg / kg body weight / day or more, 12 mg / kg body weight / day or more, 13 mg / kg body weight / day or more, 14 mg / kg body weight / day or more, 15 mg / kg body weight / day or more, 16 mg / kg body weight / day or more, 17 mg / kg body weight / day or more, 18 mg / kg body weight / day or more, 19 mg / kg body weight / day or more, 20 mg / kg body weight / day or more, 21 mg / kg body weight / day or more, 22 mg / kg body weight / day or more, 23 mg / kg body weight / day or more, 24 mg / kg body weight / day or more, calculated as monounsaturated fatty acids having 20 or more carbons. The animal may be administered at least 25 mg / kg body weight / day, at least 30 mg / kg body weight / day, at least 40 mg / kg body weight / day, at least 50 mg / kg body weight / day, at least 100 mg / kg body weight / day, or at least 200 mg / kg body weight / day of the active ingredient for 4 weeks or more, for example, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 8 weeks, at least 9 weeks, at least 10 weeks, at least 11 weeks, or at least 12 weeks. Furthermore, the ingredient selected from monounsaturated fatty acids having 20 or more carbon atoms, their salts, and their esters does not have any severe side effects, so there is no limit to the daily intake amount. EXAMPLES

[0041] The present invention will now be described in further detail with reference to examples, but the present invention is not limited to these examples. In the examples, all percentages are by weight unless otherwise specified.

[0042] Example 1: Intervention study (1) Test method a) Test food: Sanma oil capsules contain 0.35g of fish oil derived from saury, which contains LC-MUFA at a ratio of approximately 29% to total fatty acids. Blended oil capsules contain 0.35g of tuna oil and refined olive oil mixed at a weight ratio of 1:1 (containing LC-MUFA at a ratio of approximately 1.8% to total fatty acids). The total of EPA+DPA+DHA in the fatty acid composition of the saury oil capsules and blended oil capsules was 19.4% and 19.7%, respectively, which was almost the same. The fatty acid composition in the saury oil capsules and blended oil capsules is shown in Table 1. The fatty acid composition was calculated by the following method. The oil to be measured was esterified using a lower alcohol and a catalyst to obtain fatty acid lower alkyl esters. The obtained fatty acid lower alkyl esters were then analyzed using a gas chromatograph equipped with a flame ionization detector (FID). Peaks corresponding to each fatty acid were identified in the obtained gas chromatography chart, and the peak area of ​​each fatty acid was calculated using the Agilent ChemStation integration algorithm (revision C.01.03

[37] , Agilent Technologies). The fatty acid composition was calculated as the percentage of each peak area relative to the sum of the peak areas of the fatty acids. The obtained area % was equivalent to the weight % of each fatty acid in the sample. Please refer to the Japan Oil Chemists' Society (JOCS) Standard Methods for Analysis of Fats, Oils and Related Materials, 2013 Edition, 2.4.2.1-2013 Fatty Acid Composition (FID Constant Temperature Gas Chromatography) and 2.4.2.2-2013 Fatty Acid Composition (FID Programmed Temperature Gas Chromatography). The analytical conditions for gas chromatography are as follows. Equipment: Agilent 6890N GC system (Agilent) Column: DB-WAX (Agilent Technologies, 30m x 0.25mm ID, 0.25μm film thickness) Carrier gas: Helium (1.0 mL / min, constant flow) Inlet temperature: 250℃ Injection volume: 1μL Injection method: Split Split ratio: 20:1 Column oven: 180℃ - 3℃ / min - 230℃ Detector: FID Detector temperature: 250℃

[0043] [Table 1]

[0044] b) Subjects: 52 healthy adults, including students from Tokushima University, aged 20 or over, were divided into two groups (a group that took saury oil capsules and a group that took blended oil capsules). The saury oil capsule group consisted of 26 subjects with an average age of 21 (average body weight: 54.2 kg (before ingestion) and 54.8 kg (after ingestion)), while the blended oil capsule group consisted of 26 subjects with an average age of 21 (average body weight: 52.8 kg (before ingestion) and 53.0 kg (after ingestion)) (Table 2).

[0045] c) Study content: Subjects took 12 capsules per day for 4 weeks. Each of the 12 saury oil capsules contained 4.2g of saury oil, meaning that subjects took in approximately 1.1g of LC-MUFA and approximately 0.7g of n-3 polyunsaturated fatty acids (EPA+DPA+DHA) per day. Each of the 12 blended oil capsules contained 2.1g of tuna oil and 2.1g of refined olive oil, meaning that subjects took in approximately 0.07g of LC-MUFA and approximately 0.7g of n-3 polyunsaturated fatty acids (EPA+DPA+DHA) per day. Therefore, although the intake of LC-MUFA differed in each capsule group, the intake of n-3 polyunsaturated fatty acids (EPA+DPA+DHA) was approximately the same. Blood was collected at the start of the study and after 4 weeks of intake. Subjects were fasted from the night before blood collection until blood collection. Blood collected using a vacuum blood collection tube (Terumo) was left at room temperature, centrifuged at 2000 rpm for 10 minutes, and stored at -80°C until measurement. Serum TMAO was measured using an ELISA kit (HUMAN Trimethylamine-N-oxide (TMAO) ELISA KIT (Glory Science CO.,LTD)) according to the instruction manual as follows. Anti-TMAO antibody was added to purified human TMAO immobilized on a microplate and reacted. After reaction with enzyme-labeled antibody, the plate was washed and colored with TMB (3, 3', 5, 5'-tetramethylbenzidine) to measure absorbance. TMAO concentration was calculated using a calibration curve obtained from a standard product. Statistical processing was performed using the statistical software Graphpad Prism (MDF Ltd.). For comparison between two groups, an unpaired t-test was used, and for comparison within a group, a paired t-test, which is a comparison of two corresponding groups, was used. d) Implementation period: October 17th to December 7th, 2017

[0046] (2) Test results As shown in Table 2 and Figure 1, serum TMAO levels were significantly reduced in both the saury oil capsule intake group and the blended oil capsule intake group, but the saury oil capsule intake group showed a lower p value (saury oil capsule group: p = 4.3 × 10 -9 Blended oil capsule group: p = 1.0 × 10 -4 The same amount of n-3 polyunsaturated fatty acids in each capsule suggested that LC-MUFA may have a more significant effect on reducing serum levels of TMAO. Non-Patent Document 10 describes that an increase in TMAO concentration is associated with an increased risk of new major adverse cardiovascular events. In addition, a paper published in 2017 (European Heart Journal 2017 38 2948-2956) shows the results of a systematic review and meta-analysis, and describes that there is a clear association between plasma TMAO levels and increased cardiovascular risk and mortality in all subjects analyzed. Therefore, it is believed that by reducing serum TMAO levels by ingesting LC-MUFA in healthy individuals, a risk reduction or preventive effect against cardiovascular disease can be obtained.

[0047] [Table 2] [Industrial Applicability]

[0048] The present invention provides food and pharmaceutical compositions that reduce the risk of contracting diseases caused by elevated serum levels of TMAO.

Claims

1. A composition for reducing serum trimethylamine N-oxide and reducing the risk of developing chronic kidney disease caused by elevated serum levels of trimethylamine N-oxide, comprising: The composition contains as an active ingredient a component selected from monounsaturated fatty acids having 20 or more carbon atoms, salts thereof, and esters thereof, wherein the active ingredient is derived from fish oil.

2. 2. The composition according to claim 1, wherein the active ingredient is a component selected from a monounsaturated fatty acid having 20 carbon atoms, a salt thereof and an ester thereof, a component selected from a monounsaturated fatty acid having 22 carbon atoms, a salt thereof and an ester thereof, or a combination thereof.

3. 3. The composition according to claim 1, wherein the active ingredient is a glyceride containing, as a constituent fatty acid, a monounsaturated fatty acid having 20 or more carbon atoms.

4. The composition of claim 3, wherein the glyceride is a triglyceride.

5. The composition according to any one of claims 1 to 4, further comprising an n-3 polyunsaturated fatty acid and an n-6 polyunsaturated fatty acid, and the weight ratio of the n-3 polyunsaturated fatty acid to the n-6 polyunsaturated fatty acid is 4.0 or more.

6. The composition according to any one of claims 1 to 5, wherein the fish oil is saury oil.

7. The composition according to any one of claims 1 to 6, wherein the composition is used so that 2 mg / kg body weight / day or more of the active ingredient calculated as a monounsaturated fatty acid having 20 or more carbon atoms is orally administered for 4 weeks or more.

8. The composition according to any one of claims 1 to 7, which is a food composition.

9. The composition according to any one of claims 1 to 7, which is a pharmaceutical composition.

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