JP2021132646A - Production method of food and pharmaceutical composition containing unsaturated fatty acid metabolite - Google Patents

Abstract

To provide a production method of food and a pharmaceutical composition containing a new unsaturated fatty acid metabolite.SOLUTION: A production method of food and a pharmaceutical composition containing a new unsaturated fatty acid metabolite includes: mixing a first composition containing unsaturated fatty acid having two or more double bonds or its derivative with a heat-treated second composition (the second composition contains lipoxygenase but hydroperoxide lyase is inactivated), and reacting the lipoxygenase with a substance contained in the first composition; or mixing the first composition containing unsaturated fatty acid having two or more double bonds or its derivative with the second composition containing lipoxygenase, and performing heat-treatment to react the lipoxygenase with the substance contained in the first composition.SELECTED DRAWING: None

Description

The present invention relates to a method for producing a food and pharmaceutical composition containing an unsaturated fatty acid metabolite.

Protein, which accounts for about 30% of soybean (dried), contains essential amino acids in a well-balanced manner and is said to have physiological functions such as a blood cholesterol lowering effect and an obesity improving effect. In addition, soybeans contain various nutrients such as lipids, carbohydrates, dietary fiber, potassium, calcium, magnesium, iron, zinc, copper, vitamin E, vitamin B1 and folic acid. Furthermore, soy lecithin that lowers cholesterol, oligosaccharides that proliferate bifidus bacteria, soy saponin that is expected to lower blood lipids such as cholesterol, and prevent osteoporosis and improve menopausal disorders. It contains many functional substances such as isoflavones.
In fact, processed soybean products are ranked first in the "20 best health-related products that became a hot topic in 2018" selected by the Health Life Business (December 15th).
Against this background, soymilk, tofu, natto, etc., which are produced from soybeans, have recently been attracting attention as a measure against lifestyle-related diseases. Also, as is well known, many foods combined with fish rich in omega 3 fatty acids are found today. For example, a hamburger steak and tofu, a hamburger steak containing tuna, etc. (Patent Document 1 and Non-Patent Documents 1 and 2).

On the other hand, in addition to the importance of fish oil or its main components, EPA, DHA, and DPA, in recent years, remarkable anti-inflammatory effects of metabolites that bind hydroxyl groups, carbonyl groups, and epoxy groups to unsaturated fatty acids have attracted attention. Therefore, there are great expectations for their application to pharmaceuticals. It has long been known that a precursor that is a key compound for the synthesis of such metabolites, for example, 17S-hydroxydocosahexaenoic acid (17S-HDHA), is obtained by oxidation of fatty acids by soybean lipoxygenase. Interestingly, these precursors also exhibit various excellent functions such as anti-inflammatory action, anti-obesity action, cardiovascular improving action, and cranial nerve improving action.
As a recent example of this functionality, the effect of 17R-HDHA on human heat pain and arthritic pain has been reported. It was also reported that this effect is not present in Resolvins D-series and E-series, which are SPMs (Specialized proresolving lipid mediators). For 250 women with knee arthritis pain (mean age 67.4 years) who were included in this cohort study, 17S-HDHA was 162.3 pmol / mL (= 0.059 mg / L) in the serum. ), It has been reported that DHA is present at 0.14 mmol / L (= 50.9 mg / L).
SPM exhibits an extremely wide range of anti-inflammatory effects, including, for example, enteritis, peritonitis, asthma, periodontal disease, retinopathy, cerebral infarction, Alzheimer's disease, diabetes, and cancer. Therefore, research on inflammatory-converging lipid mediators is advancing day by day. During its metabolic process at the site of inflammation, one or more hydroxyl groups are enzymatically introduced into the molecule of omega-3 unsaturated fatty acids. For example, in the case of DHA, the 17th carbon is first hydroperoxylated, and the epoxy produced from this is enzymatically hydrolyzed, and at the same time, the 10th carbon is hydroxylated to Neuroprotectin D1, an anti-inflammatory lipid mediator. Is produced. In addition, resolvins D1, D2, D3, D4 and D5 are produced from 17S-HDHA, and all show an inflammation-converging effect (Non-Patent Document 3).
In addition, many papers have been published that 15-hydroxyeicosapentaenoic acid (15-HEPE) is associated with many diseases as well as 17S-HDHA. Diseases to which 15-HEPE can be applied include, for example, sputum, blood vessels, stray, influenza, kidney disease, premature birth, multiple sclerosis, sepsis, non-alcoholic fatty liver, blood-related diseases, psoriasis, diabetic skin inflammation, peritonitis. , Asthma, allergy, atopic dermatitis, depression, house dust mite asthma, psoriatic arthritis, autoimmune disease, palmoplantar keratosis, human cancer cytotoxicity, fibrosis, multiple sclerosis, And ulcerative psoriasis, but not limited to these.

From the above viewpoints, for example, foods that exhibit various physiological functions that are useful by themselves and that contain unsaturated fatty acid metabolites such as 17S-HDHA, which are precursors of resolvins and protectins, and methods for producing them are required. ing.

In addition, since such metabolites are strongly expected to be used as pharmaceutical materials, it is extremely important to develop a simple synthetic method that is low in cost and has a low environmental load (uses as few synthetic reagents as possible). Is a challenge.

Japanese Unexamined Patent Publication No. 2008-201785

Kazuhiro Uenishi et al., "Study of anti-obesity effect by ingestion of soymilk containing DHA in young Japanese adult women", Journal of Japanese Society of Nutrition and Food Science, Vol. 55, No. 6, 2002, pp. 339-345 Yukio Yamori et al., "Soy and fish as features of the Japanese diet and cardiovascular disease risks", PLOS ONE, online 2017, April 21 C. N. Serhan et al. , Resolvins: a family of bioactive products of omega-3 fatty acid transitions ititated by aspirin training. Exp. Med. , 2002, 196, 1025-1037

An object of the present invention is to provide a method for producing a food and pharmaceutical composition containing a novel unsaturated fatty acid metabolite.

For example, it is an object of the present invention to provide a method for producing a food containing unsaturated fatty acid metabolites in which a hydroxyl group and / or a hydroperoxy group is bonded to an unsaturated fatty acid.

Another object of the present invention is to provide a method for producing a pharmaceutical composition containing unsaturated fatty acid metabolites in which a hydroxyl group and / or a hydroperoxy group is bonded to an unsaturated fatty acid.

When the enzyme lipoxygenase is allowed to act on a polyunsaturated fatty acid (eg, DHA) or a derivative thereof containing two or more double bonds, useful oxides (eg, ω3 fatty acid oxide and / or ω6 fatty acid oxide) are produced. Will be done. Beans such as soybeans also contain the hydroxylase lipoxygenase. When lipoxygenase acts on unsaturated fatty acids, it results in fatty acids peroxide, which, when degraded by hydroperoxide lyases, produce aldehydes that cause a green odor. Therefore, in soybean breeding, soybeans completely lacking lipoxygenase are bred so as not to produce fatty acid peroxide. Some people use peroxidase as an enzyme that reduces a hydroperoxy group to a hydroxyl group (Osamu Furuta and Makita Haka, "Growing Lipoxygenase Completely Deleted Soybeans and Their Processing and Utilization", Jyokyo (1997), Vol. 92. , No. 8, pp. 573-578).
As described above, the problem of soybean lipoxygenase is recognized in the prior art for producing food materials (for example, wheat flour), and as a countermeasure against this problem, a gene encoding an enzyme is deleted or an enzyme protein is used. Complete inactivation was widely practiced.

On the other hand, contrary to such conventional wisdom, the present inventors have made soymilk or raw soybean milk that has been appropriately heat-treated under conditions that do not inactivate the enzyme lipoxygenase contained in soymilk. By adding a polyunsaturated fatty acid such as DHA, it was possible to prepare a food containing an anti-inflammatory metabolite (17S-hydroxydocosahexaenoic acid) produced from DHA by the enzyme lipoxygenase contained in the soymilk.

The purpose of heating is to selectively inactivate another enzyme, hydroperoxide lyase, which decomposes the hydroperoxide produced by lipoxygenase and gives an unpleasant odor. According to the present invention, highly unsaturated fatty acids (eg, EPA and DHA), which are substances that serve as substrates for the enzyme, are added to an enzyme that synthesizes a useful metabolite, for example, a living organism or food material or composition containing lipoxygenase. Then, by performing normal food processing, it has become possible to produce foods containing useful metabolites.

Generally, when lipoxygenase acts on unsaturated fatty acids, the product is an unsaturated fatty acid with an OOH group. This OOH group is reduced at the site of inflammation to form a hydroxyl group OH, which changes to a hydroxylated unsaturated fatty acid. Therefore, since the available molecular form is usually a hydroxylated unsaturated fatty acid rather than a peroxidized unsaturated fatty acid, a reaction has been carried out to reduce the OOH group to OH. Glutathione, sodium borohydride, etc. are the reducing agents.

However, surprisingly, in the unsaturated fatty acid metabolites isolated from raw gou and soymilk prepared by adding DHA from heat-treated soybean, the OOH group was already reduced to OH. This is a fact that was completely unexpected at first. The mechanism is unknown, but it is novel and provides an advantageous condition that it is not necessary to carry out a reduction reaction of OOH groups when preparing foods and pharmaceutical compositions.

The method of the present invention is characterized in that it does not require any material or method that cannot be used for food. Furthermore, it can be manufactured in the same or very close process as the normal food manufacturing process. The substance serving as a substrate may be added at the beginning of the food manufacturing process, or may be appropriately added at an intermediate point in the process. Since the useful metabolite produced by the production method of the present invention can also be used as an active ingredient of a pharmaceutical composition, the production method of the present invention is also useful as a method for producing an active ingredient of a pharmaceutical composition. Can be.

For example, the present invention provides:
(Item 1)
A method for producing an edible composition containing a metabolite of a substance consisting of an unsaturated fatty acid containing two or more double bonds or a derivative of an unsaturated fatty acid containing two or more double bonds.
(A) A step of providing a first composition containing a substance selected from the group consisting of unsaturated fatty acids containing two or more double bonds and derivatives of unsaturated fatty acids containing two or more double bonds;
(B) A step of providing a heat-treated second composition, wherein the second composition contains lipoxygenase; and (c) the first composition and the first. A step of mixing the two compositions and reacting the lipoxygenase with the substance contained in the first composition.
Including, methods.
(Item 2)
The method according to item 1, wherein the heat treatment in the step (b) is performed at 60 ° C. to 90 ° C.
(Item 3)
The method according to item 1, wherein the second composition is a bean-derived lipoxygenase-containing composition.
(Item 4)
The method according to item 1, wherein the second composition comprises a substance selected from the group consisting of soybeans, soymilk, soybean flour, soybean flour, whey, and soybean soaked liquid.
(Item 5)
The method according to item 1, wherein the second composition is a composition containing a substance derived from soybean.
(Item 6)
The method of item 1, wherein the unsaturated fatty acid is selected from the group consisting of omega-3 fatty acids and omega-6 fatty acids.
(Item 7)
The unsaturated fatty acid is selected from the group consisting of DHA, EPA, DPA (n-3), DPA (n-6), linoleic acid, α-linolenic acid, γ-linolenic acid, stearidonic acid, and arachidonic acid. , The method according to item 1.
(Item 8)
The method according to item 6, wherein the omega 3 fatty acids are EPA and DHA.
(Item 9)
The method according to item 6, wherein the omega-6 fatty acid is linoleic acid.
(Item 10)
The method of item 1, wherein the second composition comprises soybean whey.
(Item 11)
The method according to item 1, wherein the second composition comprises soybean raw gou (soybean).
(Item 12)
The method according to item 1, wherein the metabolite of the unsaturated fatty acid or the metabolite of the derivative of the unsaturated fatty acid is 17S-HDHA, 17S-HDHA ethyl ester, 17S-HDHA-bound triglyceride, 17S-HDHA-bound phospholipid. , 15-HEPE, 15-HEPE ethyl ester, 15-HEPE-linked triglyceride, 15-HEPE-linked phospholipid, 17-HDPA, 17-HDPA ethyl ester, 17-HDPA-linked triglyceride, 17-HDPA-linked phospholipid, 13-hydroxy Linolic acid, 13-hydroxylinolic acid ethyl ester, 13-hydroxylinolic acid bound triglyceride, 13-hydroxylinolic acid bound phospholipid, 13-hydroxy-α-linolenic acid, 13-hydroxy-α-linolenic acid ethyl ester, 13- Hydroxy-α-linolenic acid bond triglyceride, 13-hydroxy-α-linolenic acid bond phospholipid, 13-hydroxy-γ-linolenic acid, 13-hydroxy-γ-linolenic acid ethyl ester, 13-hydroxy-γ-linolenic acid bond Triglyceride, 13-hydroxy-γ-linolenic acid-linked phospholipid, 13-hydroxystearidonic acid, 13-hydroxystearidonic acid ethyl ester, 13-hydroxystearidonic acid-linked triglyceride, 13-hydroxystearidonic acid-linked phospholipid, 15 -Hydroxyarachidonic acid, 15-hydroxyarachidonic acid ethyl ester, 15-hydroxyarachidonic acid-bound triglyceride, 15-hydroxyarachidonic acid-bound phospholipid, 13-hydroxyoctadecadienoic acid (13-HODA), 13-hydroxyoctadecadienoic acid A method selected from the group consisting of (13-HODA) ethyl ester, 13-hydroxyoctadecadienoic acid (13-HODA) -linked triglyceride, and 13-hydroxyoctadecadienoic acid (13-HODA) -linked phospholipid.
(Item 13)
A method for producing an edible composition containing a metabolite of a substance consisting of an unsaturated fatty acid containing two or more double bonds or a derivative of an unsaturated fatty acid containing two or more double bonds.
(A) A step of providing a composition containing a lipoxygenase and a substance selected from the group consisting of unsaturated fatty acids containing two or more double bonds and derivatives of unsaturated fatty acids containing two or more double bonds. And (b) the step of heat-treating the composition,
Including, methods.
(Item 14)
The method according to item 13, wherein the heat treatment in the step (b) is performed at 60 ° C. to 90 ° C.
(Item 15)
The method according to item 13, wherein the lipoxygenase is a bean-derived lipoxygenase.
(Item 16)
13. The method of item 13, wherein the composition comprises a substance selected from the group consisting of soybeans, soymilk, soybean flour, soybean flour, whey, and soybean soaking liquor.
(Item 17)
The method according to item 13, wherein the composition is a composition containing a substance derived from soybean.
(Item 18)
13. The method of item 13, wherein the unsaturated fatty acid is selected from the group consisting of omega-3 fatty acids and omega-6 fatty acids.
(Item 19)
The unsaturated fatty acid is selected from the group consisting of DHA, EPA, DPA (n-3), DPA (n-6), linoleic acid, α-linolenic acid, γ-linolenic acid, stearidonic acid, and arachidonic acid. , Item 13.
(Item 20)
The method according to item 18, wherein the omega 3 fatty acids are EPA and DHA.
(Item 21)
The method of item 18, wherein the omega-6 fatty acid is linoleic acid.
(Item 22)
13. The method of item 13, wherein the composition comprises soybean whey.
(Item 23)
The method according to item 13, wherein the composition comprises soybean raw gou (soybean).
(Item 24)
The method according to item 13, wherein the metabolite of the unsaturated fatty acid or the metabolite of the derivative of the unsaturated fatty acid is 17S-HDHA, 17S-HDHA ethyl ester, 17S-HDHA-bound triglyceride, 17S-HDHA-bound phospholipid. , 15-HEPE, 15-HEPE ethyl ester, 15-HEPE-linked triglyceride, 15-HEPE-linked phospholipid, 17-HDPA, 17-HDPA ethyl ester, 17-HDPA-linked triglyceride, 17-HDPA-linked phospholipid, 13-hydroxy Linolic acid, 13-hydroxylinolic acid ethyl ester, 13-hydroxylinolic acid bound triglyceride, 13-hydroxylinolic acid bound phospholipid, 13-hydroxy-α-linolenic acid, 13-hydroxy-α-linolenic acid ethyl ester, 13- Hydroxy-α-linolenic acid bond triglyceride, 13-hydroxy-α-linolenic acid bond phospholipid, 13-hydroxy-γ-linolenic acid, 13-hydroxy-γ-linolenic acid ethyl ester, 13-hydroxy-γ-linolenic acid bond Triglyceride, 13-hydroxy-γ-linolenic acid-linked phospholipid, 13-hydroxystearidonic acid, 13-hydroxystearidonic acid ethyl ester, 13-hydroxystearidonic acid-linked triglyceride, 13-hydroxystearidonic acid-linked phospholipid, 15 -Hydroxyarachidonic acid, 15-hydroxyarachidonic acid ethyl ester, 15-hydroxyarachidonic acid-bound triglyceride, 15-hydroxyarachidonic acid-bound phospholipid, 13-hydroxyoctadecadienoic acid (13-HODA), 13-hydroxyoctadecadienoic acid A method selected from the group consisting of (13-HODA) ethyl ester, 13-hydroxyoctadecadienoic acid (13-HODA) -linked triglyceride, and 13-hydroxyoctadecadienoic acid (13-HODA) -linked phospholipid.
(Item 25)
An edible composition comprising a metabolite of the unsaturated fatty acid or a metabolite of a derivative of the unsaturated fatty acid produced by the method according to item 1 or 13.
(Item 26)
A method for producing a pharmaceutical composition containing a metabolite of a substance consisting of an unsaturated fatty acid containing two or more double bonds or a derivative of an unsaturated fatty acid containing two or more double bonds.
(A) A step of providing a first composition containing a substance selected from the group consisting of unsaturated fatty acids containing two or more double bonds and derivatives of unsaturated fatty acids containing two or more double bonds;
(B) A step of providing a heat-treated second composition, wherein the second composition contains lipoxygenase; and (c) the first composition and the first. A step of mixing the two compositions and reacting the lipoxygenase with the substance contained in the first composition.
Including, methods.
(Item 27)
A method for producing a pharmaceutical composition containing a metabolite of a substance consisting of an unsaturated fatty acid containing two or more double bonds or a derivative of an unsaturated fatty acid containing two or more double bonds.
(A) A step of providing a composition containing a lipoxygenase and a substance selected from the group consisting of unsaturated fatty acids containing two or more double bonds and derivatives of unsaturated fatty acids containing two or more double bonds. And (b) the step of heat-treating the composition,
Including, methods.
(Item 28)
A pharmaceutical composition comprising a metabolite of the unsaturated fatty acid or a metabolite of a derivative of the unsaturated fatty acid produced by the method according to item 26 or 27.

The present invention provides a method for producing a food containing an unsaturated fatty acid or a metabolite of a derivative thereof (eg, an oxide, more specifically, for example, an ω3 fatty acid oxide and / or an ω6 fatty acid oxide). Examples of unsaturated fatty acid metabolites include, but are not limited to, oxidative metabolites of unsaturated fatty acids.

For example, the present invention provides a method for producing a food and pharmaceutical composition containing a substance selected from the group consisting of an oxide of EPA, an oxide of DPA and an oxide of DHA. Examples of DPA include DPA (n-3) and DPA (n-6).

For example, this oxide is 17-HDHA, PDX, 17-HDPA, and 13-H.
Selected from the group consisting of SDA.
By using similar techniques (eg, by using soybeans as well), it is possible to convert unsaturated fatty acids to unsaturated fatty acid metabolites. Typical examples of unsaturated fatty acids include unsaturated fatty acids contained in foods.
An example of this is, for example
-Produce 13-hydroxylinoleic acid from linoleic acid,
Produces 13-hydroxy-α-linolenic acid from α-linolenic acid,
-Produces 13-hydroxy-γ-linolenic acid from γ-linolenic acid,
Produces 13-hydroxystearidonic acid from stearidonic acid,
Produces 15-hydroxyarachidonic acid from arachidonic acid,
-Generate 17-HDPA from DPA,
• Producing 15-HEPE from EPAs, but is not limited to these. The unsaturated fatty acid metabolite may be a derivative such as an ethyl ester, a conjugate with a triglyceride, or a conjugate with a phospholipid.

In the present invention, not only unsaturated fatty acids but also unsaturated fatty acid derivatives such as polyunsaturated fatty acid derivatives can be used as substrates.

Derivatives of highly unsaturated fatty acids include, for example, ethyl ester of highly unsaturated fatty acid, triglyceride containing highly unsaturated fatty acid, diglyceride containing highly unsaturated fatty acid, monoglyceride containing highly unsaturated fatty acid, and phosphorus containing highly unsaturated fatty acid. Lipids, lysoline lipids containing highly unsaturated fatty acids, sphingolipids containing highly unsaturated fatty acids, and glycolipids containing highly unsaturated fatty acids, and amide derivatives of highly unsaturated fatty acids (eg, ethanol of highly unsaturated fatty acids). Fatty acid amides such as amides), but are not limited to these.

Derivatives of polyunsaturated fatty acids also include esters of polyunsaturated fatty acids. Unsaturated fatty acid esters (eg, unsaturated fatty acid ethyl esters), in particular highly unsaturated fatty acid esters (eg, highly unsaturated fatty acid ethyl esters), are also available as substrates for enzymatic reactions.

In the production of foods and / or pharmaceutical compositions containing unsaturated fatty acid metabolites, derivatives of highly unsaturated fatty acids (eg, EPA, DPA or DHA) are free fatty acid-type, alcohol-type, ester-type and other physiologically active substances. Complex with, various glyceride types (eg, monoglyceride, diglyceride, triglyceride, alkyl glyceryl ether lipid, alkenyl glyceryl ether lipid), various phospholipid types (eg, glycerophospholipid, sphingolin lipid), various amine salts (eg, trimethylamine) Aliphatic amine salts such as salts, triethylamine salts, dicyclohexylamine salts, ethanolamine salts, diethanolamine salts, triethanolamine salts, procaine salts, meglumine salts, diethanolamine salts or ethylenediamine salts; N, N-dibenzylethylenediamine salts, venetamine salts. Aralkylamine salts such as pyridine salts, picolin salts, quinoline salts, isoquinolin salts and other heterocycle aromatic amine salts), various ammonium salts (eg, tetramethylammonium salt, tetraethylammonium salt, benzyltrimethylammonium salt, benzyltriethylammonium salt) Salts, quaternary ammonium salts such as benzyltributylammonium salt, methyltrioctylammonium salt, tetrabutylammonium salt) and various metal salts (eg, lithium, sodium, potassium, cesium, rubidium, calcium, magnesium, aluminum, tin , Zinc, lead, cobalt, nickel salts). In addition, the alcohol moiety in the ester form is of methanol, ethanol, propanol, isopropanol, butanol and its isomers, pentanol and its isomers, hexanol and its isomers, heptanol and its isomers, octanol and its isomers, and more molecular weight. It is selected from the group consisting of large alcohols (eg, oleyl alcohols) and their isomers, as well as alcohols containing unsaturated bonds and their isomers.

Compositions comprising highly unsaturated fatty acids or oxides of derivatives thereof (eg, ω3 fatty acid oxides and / or ω6 fatty acid oxides) containing two or more double bonds of the present invention also improve insulin resistance (eg, ω3 fatty acid oxides and / or ω6 fatty acid oxides). Treatment and prevention), insulin sensitivity enhancement, diabetes prevention and treatment, glucose metabolism improvement, low density lipoprotein reduction, cholesterol reduction, obesity reduction, hypertriglyceridemia prevention and treatment, atheroscleridemia prevention and treatment Shows excellent effects in such cases.

The present invention provides inflammatory converging metabolites (eg, 17-hydroxydocosahexaenoic acid) of polyunsaturated fatty acids or derivatives thereof containing two or more double bonds, which are novel and have superior advantages over prior art. Provide a method of producing a food containing. Since the useful metabolite produced by the production method of the present invention (for example, an inflammatory converging metabolite) can also be used as an active ingredient of a pharmaceutical composition, the production method of the present invention is a pharmaceutical composition. It can also be useful as a method for producing the active ingredient of.

The present invention is not a general method of synthesizing such a metabolite from an omega-3 fatty acid such as docosahexaenoic acid and adding the metabolite to a food, but an anti-inflammatory omega 3 fatty acid metabolite during food manufacturing and processing. It is produced and accumulated, and as a result, it becomes possible to produce foods containing such useful substances. Since the useful metabolites produced by the production method of the present invention (for example, anti-inflammatory ω3 fatty acid metabolites and / or ω6 fatty acid oxides) can also be used as active ingredients of pharmaceutical compositions, the present invention. The method for producing the above can also be useful as a method for producing an active ingredient of a pharmaceutical composition.

Hereinafter, the present invention will be described. Throughout the specification, it should be understood that the singular representation also includes its plural concept, unless otherwise stated. It should also be understood that the terms used herein are used in the meaning commonly used in the art unless otherwise noted. Thus, unless otherwise defined, all terminology and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, this specification (including definitions) takes precedence. Also, in the present specification, "wt%" is used interchangeably with "mass fraction concentration". “%” Means “wt%” or “w / w%” or “mass percent concentration” unless otherwise specified.
The following is a list of definitions of terms specifically used herein.

As used herein, the term "unsaturated fatty acid containing two or more double bonds" refers to any unsaturated fatty acid containing two or more double bonds. It is preferably a fatty acid containing a double bond at the ω3 position and other positions, and more preferably a fatty acid containing a double bond at the ω3 position and the ω6 position. The oxide of the unsaturated fatty acid of the present invention is preferably a fatty acid in which one or more hydroxyl groups are bonded at a specific position. Typical unsaturated fatty acids containing two or more double bonds include hexadecatolic acid, hexadecatetraenoic acid, linolenic acid, α-linolenic acid, γ-linolenic acid, stearidonic acid, and dihomo-γ. -Linolenic acid, eikosatetraenoic acid, arachidonic acid, eicosapentaenoic acid (EPA), docosapentaenoic acid, docosapentaenoic acid, docosapentaenoic acid, docosapentaenoic acid (DPA) (DPA is DPA (n-3) and DPA (n-6)), docosapentaenoic acid (DHA), tetracosapentaenoic acid, and tetracosapentaenoic acid, respectively, refer to all fatty acids including n-3 and n-6 series. Is not limited to these. Examples of the unsaturated fatty acid oxide (modified product) containing two or more double bonds include 15-hydroxyeicosatetraenoic acid, 19-hydroxytetracosapentaenoic acid, and 19-hydroxytetracosahexahexa. Enic acid, 11-hydroxyhexadecatrienoic acid, 11-hydroxyhexadecatetraenoic acid, 13-hydroxylinoleic acid, 13-hydroxy-α-linolenic acid, 13-hydroxy-γ-linolenic acid, 13-hydroxystearidonic acid , 15-Hydroxydihomo-γ-linolenic acid, 15-hydroxyeicosatetraenoic acid, 15-hydroxyEPA, 17-hydroxydocosadienoic acid, 17-hydroxydocosatrienoic acid, 17-hydroxydocosatetraenoic acid, 17-hydroxy DPA, 17-Hydroxy DHA, 19-Hydroxytetracosapentaenoic acid, 19-Hydroxytetracosahexaenoic acid, Hexadecadienoic acid (C16: 2), Hexadecatrienoic acid (C16: 3), Hexadecatetraenoic acid (C16: 4), Eikosazienoic acid (C20: 2), Docosazienoic acid (C22: 2), Pinolenic acid (C18: 3), Eleostearic acid (C18: 3), Meadic acid (C20: 3), Adren ( C22: 4), boseopentaenoic acid (C18: 5), osbondic acid (C22: 5), sardine acid (C22: 5), tetracosapentaenoic acid (C24: 5), and heric acid (C24: 6) These include, but are not limited to.

As used herein, the term "derivat of unsaturated fatty acid containing two or more double bonds" refers to a derivative of any unsaturated fatty acid containing two or more double bonds. Unsaturated fatty acids are preferably fatty acids that contain a double bond at the ω3 and other positions, and more preferably a double bond at the ω3 and ω6 positions. Derivatives of highly unsaturated fatty acids include free fatty acid type, alcohol type, ester type, complex with other physiologically active substances, and various glyceride types (for example, monoglyceride, diglyceride, triglyceride, alkylglyceryl ether lipid, alkenylglyceryl ether lipid). , Various phospholipid types (eg, glycerophospholipid, sphingolinlipid), various amine salts (eg, trimethylamine salt, triethylamine salt, dicyclohexylamine salt, ethanolamine salt, diethanolamine salt, triethanolamine salt, procaine salt, meglumin salt, An aliphatic amine salt such as a diethanolamine salt or an ethylenediamine salt; an aralkylamine salt such as an N, N-dibenzylethylenediamine salt or a venetamine salt; a heterocycle aromatic amine salt such as a pyridine salt, a picolin salt, a quinoline salt or an isoquinolin salt), Various ammonium salts (for example, quaternary ammonium salts such as tetramethylammonium salt, tetraethylammonium salt, benzyltrimethylammonium salt, benzyltriethylammonium salt, benzyltributylammonium salt, methyltrioctylammonium salt, tetrabutylammonium salt) and It may be selected from the group consisting of various metal salts (eg, salts of lithium, sodium, potassium, cesium, rubidium, calcium, magnesium, aluminum, tin, zinc, lead, cobalt, nickel). In addition, the alcohol moiety in the ester form is of methanol, ethanol, propanol, isopropanol, butanol and its isomers, pentanol and its isomers, hexanol and its isomers, heptanol and its isomers, octanol and its isomers, and more molecular weight. It is selected from the group consisting of large alcohols (eg, oleyl alcohols) and their isomers, as well as alcohols containing unsaturated bonds and their isomers.

As used herein, the term "a metabolite of a substance consisting of an unsaturated fatty acid containing two or more double bonds or a derivative of an unsaturated fatty acid containing two or more double bonds" is defined as a double bond. An unsaturated fatty acid containing two or more or a derivative of an unsaturated fatty acid containing two or more double bonds is treated with an enzyme such as lipoxygenase, and an enzymatic reaction product such as an oxide obtained thereby is treated. To say.

The term "ω3 fatty acid" used in the present specification refers to an unsaturated fatty acid having a double bond from the third carbon (ω3 position) from the methyl terminal, and typically α-linolenic acid and stearidone. Acids, eicosatetraenoic acid, eicosapentaenoic acid (EPA), docosapentaenoic acid (DPAn-3), docosapentaenoic acid (DHA), tetracosapentaenoic acid, and tetracosapentaenoic acid. Not limited.
As used herein, the term "ω6 fatty acid" refers to an unsaturated fatty acid that has a double bond from the 6th carbon (ω6 position) from the methyl terminal, and is typically docosapentaenoic acid (DPAn). -6), arachidonic acid, γ-linolenic acid, and linoleic acid, but are not limited thereto.

As used herein, the term "ω3 fatty acid metabolite" is a substance obtained by reacting ω3 fatty acid with a metabolic enzyme, and includes an oxide of ω3 fatty acid (for example, an oxidative metabolite). Typically, 13-hydroxy-α-linolenic acid, 13-hydroxystearidonic acid, 15-hydroxyeicosatetraenoic acid, 15-hydroxyEPA (15-HEPE), 17-hydroxyDPan-3, 17-hydroxy Examples include, but are not limited to, DHA, 19-hydroxytetracosapentaenoic acid, and 19-hydroxytetracosahexaenoic acid, and their ethyl esters, triglyceride conjugates, and phospholipid conjugates.
As used herein, the term "ω6 fatty acid metabolite" is a substance obtained by reacting ω6 fatty acid with a metabolic enzyme, and includes an oxide of ω6 fatty acid (for example, an oxidative metabolite). Typically, 15-hydroxyeicosatetraenoic acid (n-6), 17-hydroxy DPan-6, 13-hydroxy-γ-linolenic acid, and 13-hydroxyoctadecadienoic acid (13-HODA), In addition, these ethyl esters, triglyceride conjugates, and phospholipid conjugates are included, but are not limited to these.

As used herein, the term "glyceride" includes components selected from the group consisting of triglycerides, diglycerides, and monoglycerides of fatty acids. In the present invention, "glyceride" does not include phospholipids or glycolipids unless otherwise specified.

The term "docosahexaenoic acid" as used herein is used interchangeably with "DHA" and is ester-bonded to the form of free fatty acids and lower alcohols such as ethanol, propanol, glycerin and phospholipids. Includes any form.

As used herein, the term "17-HDHA" or "17S-HDHA" is an oxide of DHA, which means DHA in which the 17th position is oxidized and "-OH" is bound. The term "17-HDHA" is used interchangeably with the term "17-hydroxy DHA" and the term "17-hydroxydocosahexaenoic acid".

The term "eicosapentaenoic acid" as used herein is used interchangeably with "EPA" and in the form of free fatty acids and lower alcohols such as ethanol, propanol, lower alcohols such as propanol, etc. It includes both glycerin and phospholipid ester-bonded forms.

The term "docosapentaenoic acid" as used herein is used interchangeably with "DPA" and is in the form of a free fatty acid and in the form ester-bonded to lower alcohols such as ethanol, propanol and glycerin. Both are included. Also, "DPA" is "n-3 DPA" (ie, all-cis-7,10,13,16,19-docosapentaenoic acid) or "n-6 DPA" (ie, all-cis). It means -4,7,10,13,16-docosapentaenoic acid). "N-3 DPA" is used interchangeably with "DPA (n-3)" and / or "ω-3 DPA". "N-6 DPA" is used interchangeably with "DPA (n-6)" and / or "ω-6 DPA". DPA includes all forms of free fatty acids, lower alcohols such as ethanol and propanol, lower alcohols such as propanol, and forms ester-bonded to glycerin.

As used herein, the term "17-HDPA" refers to an oxide of DPA, a DPA in which the 17th position is oxidized and "-OH" is bound.

As used herein, the term "stearidonic acid" is used interchangeably with "SDA" and can be in the form of a free fatty acid, lower alcohols such as ethanol, propanol, or ester-bonded to glycerin. Also includes.

The term "α-linolenic acid" as used herein is used interchangeably with "ALA" and is in the form of a free fatty acid and in the form of ester-bonded to lower alcohols such as ethanol, propanol, glycerin. Including any of.

The term "eicosatetraenoic acid" as used herein is used interchangeably with "ETA" and is ester-bonded to the form of free fatty acids and lower alcohols such as ethanol, propanol, glycerin. Includes any form.
As used herein, the term "linoleic acid" is used interchangeably with "LA" and can be in the form of a free fatty acid, lower alcohols such as ethanol, propanol, or ester-bonded to glycerin. Also includes.
The term "arachidonic acid" as used herein is used interchangeably with "AA" or "ARA" and is ester-bonded to the form of free fatty acids and lower alcohols such as ethanol, propanol, glycerin. Includes any of the above forms.

As used herein, the term "lipoxygenase" refers to an oxidoreductase that introduces a hydroperoxy group by adding molecular oxygen to an unsaturated fatty acid. The term "lipoxygenase" is used interchangeably with "lipoxygenase". Lipoxygenase is widely found in plants. Sources of lipoxygenase include, for example, beans (eg, beans selected from the group consisting of soybeans, green beans, tiger beans, quail beans, and green peas), potatoes, tomatoes, cabbage, and white bean. Not limited to. In the present invention, a bean-derived lipoxygenase-containing composition can be used to carry out an oxidation reaction with lipoxygenase.

As used herein, the term "lipoxygenase-containing composition" is a composition (liquid or solid) prepared from a plant material (whole plant or part of a plant) containing lipoxygenase, and is a plant as a raw material. A composition containing a component other than lipoxygenase contained in the material and at least one lipoxygenase contained in the plant material as a raw material.
Sources of lipoxygenase and lipoxygenase-containing compositions include, for example, legumes such as soybeans, as well as, for example, lipoxygenase-containing marine organisms (fish, shrimp and crab crustaceans, algae, etc.) and plants (eg barley, barley, etc.). Tomatoes, potatoes, etc.), but are not limited to these.

The "bean-derived lipoxygenase-containing composition" is a composition (liquid or solid) containing a substance prepared from beans, and contains lipoxygenase contained in the raw material beans and the raw material beans. A composition containing a component other than at least one lipoxygenase. The content of the component other than at least one lipoxygenase contained in the plant material (for example, beans) as a raw material in the composition is, for example, 0.5% (w / w) or more and 1% (w / w /). w) or more, 2% (w / w) or more, 3% (w / w) or more, 4% (w / w) or more, 5% (w / w) or more, 6% (w / w) or more, 7 % (W / w) or more, 8% (w / w) or more, 9% (w / w) or more, 10% (w / w) or more, 12% (w / w) or more, 14% (w / w) ) Or more, 16% (w / w) or more, 18% (w / w) or more, or 20% (w / w) or more, but is not limited thereto. Compositions prepared from plant materials (eg, beans) may be liquid, solid, or both liquid and solid. Compositions prepared from beans include, but are not limited to, for example, bean flour and bean soaking liquor, and any composition containing them.

As used herein, the term hydroperoxide lyase refers to an enzyme that oxidatively cleaves the peroxide structure and breaks it down into two hydroxyl groups.

As used herein, the term "bean flour" refers to flour obtained by crushing beans. Examples of beans include, but are not limited to, beans selected from the group consisting of soybeans, kidney beans, tiger beans, quail beans, and green peas. The conditions for crushing are not particularly limited, and the particle size of the crushed beans is also not particularly limited. For example, from crushed beans having a particle size of 2000 μm or more, crushed products of 600 μm to 2000 μm, 150 μm to 600 μm, or 150 μm or less can be used. In the present invention, the soybean flour is preferably soybean flour.

As used herein, the term "soybean flour" refers to crushed soybean flour.

As used herein, the term "bean soaking solution" refers to a solution obtained by immersing beans in a liquid (eg, water). Examples of beans include, but are not limited to, beans selected from the group consisting of soybeans, kidney beans, tiger beans, quail beans, and green peas. The liquid may be aqueous or oily, preferably aqueous. As the aqueous liquid, any liquid such as water and a buffer solution can be used. The immersion conditions are not particularly limited, and any conditions can be used as long as a bean immersion solution exhibiting lipogycigenase activity can be obtained. In the present invention, the bean soaking solution is preferably a soybean soaking solution.

The term "raw soybean" used in the present specification refers to soybeans being immersed in an aqueous solution such as water (the immersion water may be removed or replaced as necessary) and appropriately heated. A suspension of a pulverized product prepared by pulverizing after processing. In addition, soymilk can be produced by further heat-treating raw gourd, if necessary, and filtering it with a cloth or the like. In addition, raw gou may be produced by using other beans instead of soybean as a source of lipogycigenase.
As used herein, the term "soaked soybean" refers to a substance in which soybeans are immersed in an aqueous solution such as water without heat treatment. For this soaking, it is common to put the soybeans in water and leave them overnight. The "water" obtained by removing soybeans from the "soaked soybeans" is the "soybean soaking liquid" as the "bean soaking liquid". When producing "immersed soybeans", the immersion water may be removed or replaced as needed. Further, if necessary, the soybeans soaked may be ground, or a suspension of such ground products may be used. In addition, "raw soybean" is obtained by crushing "soaked soybean" with a mixer or the like, and "soy milk" is a filtrate obtained by adding water to "raw soybean", heating it, and filtering it with a cloth. You can also add bittern to soymilk to make tofu.

(Enzyme reaction using lipoxygenase-containing composition)
The lipoxygenases that can be used in the present invention are widely contained in plants. Sources of lipoxygenase include, for example, beans (eg, beans selected from the group consisting of soybeans, green beans, tiger beans, quail beans, and green peas), tomatoes, potatoes, cabbage, and white bean. Not limited to.

For example, in the present invention, a lipoxygenase-containing composition (for example, a bean-derived lipoxygenase-containing composition) can be used to carry out a lipoxygenase enzyme reaction. Examples of the bean-derived lipoxygenase-containing composition include, but are not limited to, soaked soybean, soybean flour, bean soaking solution, raw gou (namago), whey (for example, soybean whey), and soymilk.

Examples of beans used as raw materials include, but are not limited to, beans selected from the group consisting of soybeans, green beans, tiger beans, quail beans, and green peas. Typical examples of soybeans used for preparing soybean flour or soybean soaking solution used in the present invention include, but are not limited to, soybeans and seeds produced by legumes. Examples of soybeans to be used include, but are not limited to, colored soybeans such as blue soybeans and black soybeans in addition to general yellow soybeans.

The crushing conditions for preparing bean flour from beans are not particularly limited. The particle size of the crushed bean flour is also not particularly limited. For example, from crushed beans having a particle size of 2000 μm or more, crushed products of 600 μm to 2000 μm, 150 μm to 600 μm, or 150 μm or less can be used.

The grinding conditions for preparing soybean flour from soybean are also not particularly limited. The particle size of the crushed soybean powder is also not particularly limited. For example, from crushed soybean having a particle size of 2000 μm or more, crushed product of 600 μm to 2000 μm, 150 μm to 600 μm, or 150 μm or less can be used.

The crushing conditions for preparing raw soybeans from soybeans are also not particularly limited. The particle size of the crushed particles is also not limited.

Enzyme reaction conditions using a lipoxygenase-containing composition (eg, soybean-derived lipoxygenase-containing composition such as soaked soybean, soybean flour, bean soaking solution, raw gou (namego), whey (for example, soybean whey), or soymilk) Can be used under any reaction conditions under which the lipoxygenase enzyme contained in the lipoxygenase-containing composition exhibits activity. Examples of such enzyme reaction conditions include, but are not limited to, the following. For example, the bean-derived lipoxygenase-containing composition includes, but is not limited to, soaked soybean, soybean flour, bean soaking solution, raw gou (namago), whey (for example, soybean whey), and soymilk. Alternatively, soaked beans, soybean flour, bean soaking solution, raw gou (namago), whey, and soymilk prepared from beans other than soybean can be used. The non-limiting reaction conditions are as follows:
(1) Buffer solution to be used: Boric acid buffer pH 9
(2) Temperature to be used: Room temperature (3) Concentration of lipoxygenase-containing composition to be used (for example, bean-derived lipoxygenase-containing composition): Typically 5% (weight / weight) to 20% (weight / weight) (4) The ratio of the substrate to the lipoxygenase-containing composition (for example, the bean-derived lipoxygenase-containing composition): the ratio of the substrate (for example, DHA free fatty acid) to the lipoxygenase-containing composition is 2% or less is desirable, but not limited to this. For example, the reaction is possible with 0.1% to 1% or 1% to 5%, 5% or more. (5) Reaction time: 0.5 to 5 hours. Reaction is possible with more or less

For example, when a crushed bean product (for example, crushed soybean product) is added, the crushed bean product can be easily removed from the reaction solution after the reaction is completed by adding the crushed bean product using a mesh net bag or a basket-shaped container. Alternatively, when a bean soaking solution (for example, soybean soaking solution) is added, since the crushed product has been removed from the bean soaking solution in advance, the step of removing the crushed bean product from the reaction solution may be omitted after the reaction is completed. can.

(Manufacturing of food and drink composition)
(Heat treatment)
The hydroperoxide molecules produced during soybean immersion grinding are immediately degraded by the hydroperoxide degrading enzyme hydroxylyase coexisting in soybeans, producing the well-known green odor. Therefore, not only the green odor but also the hydroperoxide production yield is greatly reduced. However, hydroxylyase is more sensitive to heat than lipoxygenase and is easily decomposed by heating at around 70 ° C. (Non-Patent Document 5). Based on this property, soybeans soaked in water are heated at 70 ° C. for about 7 minutes, then crushed into a suspension, and an appropriate amount of free fatty acid of DHA is added to the suspension, stirred, filtered, and then made into soymilk. A soybean food containing 17S-HDHA can be obtained by further adding bittern to make tofu. Further, the soybeans before soaking may be heated in the same manner. (Non-Patent Documents 4 and 5) In general soymilk preparation, the suspension is filtered through cotton and then heated to about 100 ° C. The purpose of this is to decompose toxic substances contained in raw soybeans. To do. Heating at about 70 ° C in this patent is not for this purpose (Kazuo Shibasaki, Shunichi Fukano, Kazuyoshi Okubo, "Food Chemistry Research on Soy Protein (14th Report) Heat Deactivation of Lipoxygenase (Part 1)", Journal of Food Science, Vol. 20, No. 9, 415-420, (1973), and B. Gini and RB Koch, Study of a lipohydroperoxide breakdown factor in soyextracts, Journal of food Science, 26, 359-364, ( 1961)).

As used herein, the term "heat treatment" refers to a condition that more inactivates hydroperoxide lyase (an enzyme that catalyzes the degradation and aldehyde formation of peroxides produced by lipoxygenase) than lipoxygenase, ie, lipoxygenase. The condition is that the level of inactivation of hydroperoxide lyase is higher than the level of inactivation. Preferably, the treatment does not inactivate lipoxygenase, but satisfies the condition of inactivating the hydroperoxide lyase that catalyzes the decomposition and aldehyde production of the peroxide produced by lipoxygenase. When the heat treatment of the present invention is performed on a lipoxygenase-containing composition, 50% or more, 60% or more, 70% or more, 80% or more, 85% or more, 90% as compared with the enzyme activity before the heat treatment. As mentioned above, 95% or more, or 98% or more of lipoxygenase activity remains. When the heat treatment of the present invention is performed on a lipoxygenase-containing composition, 50% or less, 40% or less, 30% or less, 25% or less, 20% or less, 15% as compared with the enzyme activity before the heat treatment. Below, the hydroperoxide lyase activity is reduced to 10% or less, or 5% or less.

Typical heat treatment is not limited, but is, for example, 40 ° C. to 90 ° C., preferably 60 ° C. to 90 ° C., more preferably 65 ° C. to 75 ° C., and most preferably 70 ° C.
The heat treatment time is not limited, but is, for example, 10 seconds to 30 minutes, preferably 5 minutes to 20 minutes, more preferably 6 minutes to 17 minutes, and even more preferably 6 minutes to 10 minutes. Is 7 minutes.

Another notable and important fact that was totally unexpected is that the product from DHA added during heat treatment and grinding is 17-hydroperoxy-DHA, as is the case with normal lipoxygenase reactions. Contrary to the expectation, as described in Examples, as a result of isolating and confirming the structure of the product, it was found that it was a hydroxy group instead of a hydroperoxy group. Since it is a well-known fact that the oxidation product of lipoxygenase is hydroperoxide, the hydroperoxide (17S-HpDHA) once produced in the heat-treated and crushed soybean sample is hydroxylated (17S-HDHA) for an unknown reason. ) Has been converted.

From a food application perspective, this fact is a very useful result. This indicates that there is no need to convert the primary product of lipoxygenase, hydroperoxide, to inflammation-convergent 17S-HDHA. I didn't expect this at all. Generally, one of the amino acids, cysteine, and sodium borohydride and triphenylphosphine, which cannot be used in foods, must be used for this conversion.

In addition to these facts, the above heat treatment has made it possible for the first time to prepare foods containing inflammatory-convergent unsaturated fatty acid metabolites by a novel method of selectively heat-inactivating unwanted enzymes. Soymilk containing the inflammatory converging metabolite 17S-HDHA is the first example, and not only the preparation method is novel, but soymilk containing the inflammatory converging metabolite can be said to be the first novel soybean food.

(Food and drink composition)
A preferred embodiment of the present invention is a food and drink composition. That is, the pharmaceutical composition or edible composition containing the ω3 fatty acid metabolite and / or ω6 fatty acid oxide of the present invention as an active ingredient is a liquid, gel-like or solid food as it is, for example, a juice or a refreshing beverage. , Coffee, tea, Japanese tea, oolong tea, vegetable juice, natural fruit juice, dairy drink, milk, soy milk, sports drink, near water drink, nutritional drink, coffee drink, cocoa, soup, dressing, mousse, jelly, yogurt, Pudding, sprinkle, powdered milk for childcare, processed milk, sports drinks, nutritional drinks, cake mix, bread, pizza, pie, crackers, biscuits, cakes, cookies, spaghetti, macaroni, pasta, udon, buckwheat, ramen, candy, soft candy , Gum, chocolate, okaki, potato chips, snacks, ice cream, sherbet, cream, cheese, powdered milk, condensed milk, dairy products such as milk drinks, buns, buns, corn, rice cakes, rice cakes, soy sauce, sauce, noodles Soup, sauce, dashi-no-moto, stew-no-moto, soup-no-moto, compound seasoning, curry-no-moto, mayonnaise, ketchup, retort curry, retort stew, retort soup, retort bowl, canned food, ham, hamburger, meat ball, croquette , Dumplings, pilafs, rice balls, frozen and refrigerated foods, chikuwa, gamo, bento rice, sushi, baby milk, baby foods, baby foods, sports foods, nutritional supplements, supplements, health foods, etc. Depending on the situation, it can be processed into pellets, tablets, granules, etc. together with moldants such as dextrin, lactose, starch, fragrances, pigments, etc., or it can be coated with gelatin etc. and molded into capsules for health foods, nutritional supplements, etc. Can be used as.

The blending amount of the ω3 fatty acid oxide and / or the ω6 fatty acid oxide in these foods or food and drink compositions is not limited, but is, for example, 1% (w / w) to 60% (w / w). ), 3% (w / w) to 50% (w / w), 5% (w / w) to 40% (w / w), 10% (w / w) to 30% (w / w), Alternatively, it is 15% (w / w) to 25% (w / w). The blending amount of DHA in these foods or food and drink compositions is not limited, but is, for example, 1% (w / w) to 80% (w / w) to 10% (w / w) to. It is 70% (w / w), 20% (w / w) to 60% (w / w), or 30% (w / w) to 55% (w / w). The amount of the unsaturated fatty acid oxide compounded when used in supplements and the like is typically 0.1% to 10% or more.

When the composition of the present invention is used as a supplement, the active ingredient may be used as it is, or it may be formulated and administered in a soft capsule, a tablet, a powder, a granule or the like. When the composition of the present invention is used in foods and drinks, it may be added to the raw materials of foods and drinks and administered.

As mentioned above, the activity of lipoxygenase is effectively utilized by heating hydroperoxide lyase, which gives an unfavorable result because it decomposes hydroperoxide, to around 70 ° C immediately after soybean immersion, and as an unexpected result, hydro. It was clarified that soymilk containing 17S-HDHA can be prepared more efficiently without the need to reduce the peroxide to a hydroxyl group. This result is also effective in the conventional case of converting unsaturated fatty acids such as DHA into hydroperoxide using soymilk or soybean flour for purposes other than food, and more efficiently produces 17S-HDHA. You should be able to prepare.
(Heat treatment)
The heat treatment of the present invention is more inactivating hydroperoxide lyase (an enzyme that catalyzes the decomposition and aldehyde production of peroxide produced by lipoxygenase) than lipoxygenase, for example, the level of inactivation of lipoxygenase. A condition in which the level of inactivation of hydroperoxide lyase is high, that is, a condition in which the inactivation rate of hydroperoxide lyase is higher than the inactivation rate of lipoxygenase. Preferably, the treatment does not inactivate lipoxygenase, but satisfies the condition of inactivating the hydroperoxide lyase that catalyzes the decomposition and aldehyde production of the peroxide produced by lipoxygenase. When the heat treatment of the present invention is performed on a lipoxygenase-containing composition, 50% or more, 60% or more, 70% or more, 80% or more, 85% or more, 90% as compared with the enzyme activity before the heat treatment. As mentioned above, 95% or more, or 98% or more of lipoxygenase activity remains. When the heat treatment of the present invention is performed on a lipoxygenase-containing composition, 50% or less, 40% or less, 30% or less, 25% or less, 20% or less, 15% as compared with the enzyme activity before the heat treatment. Below, the hydroperoxide lyase activity is reduced to 10% or less, or 5% or less.
The heating conditions used in the present invention are, for example, conditions for heating an enzyme mixture containing lipoxygenase and hydroperoxide lyase, and when the heated enzyme mixture is used, 17S- in an enzymatic reaction using DHA as a substrate. This is a heating condition that maximizes HDHA production. Alternatively, the heating conditions used in the present invention are, for example, conditions for heating an enzyme mixture containing lipoxygenase and hydroperoxide lyase, in which the heated enzyme mixture is used as a substrate with eicosapentaenoic acid (EPA). 13-Hydroxyoctadecadienoic acid (13-) in the heating conditions that maximize the production of 15-hydroxyeicosapentaenoic acid (15-HEPE) in the enzymatic reaction, or in the enzymatic reaction using linoleic acid (LA) as a substrate. HODA) Heating conditions that maximize production. As a method for measuring the production amount of 17S-HDHA (or 15-HEPE or 13-HODA, etc.), for example, a lipid component containing DHA (or EPA or LA) (for example, a lipid component contained in soy milk) is used as a substrate. Quantitative methods include enzymatic reaction using, analysis by thin layer chromatography (TLC), and cochromatography with 17S-HDHA (or 15-HEPE, 13-HODA, etc.) preparations. In this method, a sample obtained by reacting a sample of an enzyme mixture to be measured (for example, soybean flour, soybean soaking solution, soybean flour, and soymilk, but not limited to these) is prepared at a known concentration. Developed by TLC with an ethanol solution (standard) of 17S-HDHA (or 15-HEPE or 13-HODA, etc.) and based on the relative spot intensity, 17S-HDHA (or 15-HEPE or 13-) contained in the sample. Quantify HODA, etc.). Alternatively, a spectrophotometer is used instead of TLC, and a quantification method using the absorption intensity at 235 nm common to the metabolite structure, or quantification by HPLC is also possible.
In another aspect of the invention, the "heat treatment" of the invention (1) reduces peroxidase (hydroperoxidase) activity to 10-30% or less, preferably 1-10% or less before heating, and Conditions that do not reduce the lipoxygenase activity to 20% or less, preferably 20 to 50% or less before heating, or (2) the ratio of lipoxygenase activity (unit) to peroxidase (hydroperoxidase) activity (unit) after heating. The condition is 5 to 10 or more, more preferably 10 to 20 or more, and most preferably 20 or more.
The production method of the present invention is a method for producing a composition containing a metabolite of a substance consisting of an unsaturated fatty acid containing two or more double bonds or a derivative of an unsaturated fatty acid containing two or more double bonds. a) Provided is a first composition comprising a substance selected from the group consisting of unsaturated fatty acids containing two or more double bonds and derivatives of unsaturated fatty acids containing two or more double bonds (b). ) A heat-treated second composition containing lipoxygenase can be provided and carried out by a method comprising (c) mixing these first and second compositions to carry out a lipoxygenase enzymatic reaction. can.
Alternatively, the production method of the present invention is selected from the group consisting of (a) lipoxygenase and an unsaturated fatty acid containing two or more double bonds and a derivative of an unsaturated fatty acid containing two or more double bonds. It can also be carried out by providing a composition comprising a substance and then (b) heat treating the composition. That is, when the mixture containing the enzyme and the substrate is heat-treated, the lipoxygenase enzyme reaction can be carried out at the same time. Further, in this embodiment, after the heat treatment, the (c) lipoxygenase enzyme reaction may be further carried out under a temperature condition different from that of the heat treatment of (b), for example. In this case, as the temperature at which the lipoxygenase enzyme reaction of (c) is carried out, an appropriate temperature for the target lipoxygenase is appropriately selected.
(Manufacturing of pharmaceutical compositions)

Further, a production method using a composition produced by the above-mentioned method for producing a food and drink composition (for example, an enzyme (enzyme-containing extract) treated by heat treatment), and a heat treatment of a composition containing an enzyme and a substrate. The production method including) is also effective in producing a pharmaceutical composition.

The present invention is a composition comprising a metabolite of an unsaturated fatty acid containing two or more double bonds (eg, ω3 fatty acid oxide and / or ω6 fatty acid oxide) for the prevention, treatment and insulin sensitivity of diabetes. Provided are excellent compositions for enhancing, improving glucose metabolism, low density lipoproteins, reducing cholesterol, and preventing and treating obesity, hypertriglyceridemia, atherosclerosis, and related diseases.

The pharmaceutical compositions of the present invention include, for example, treatment and prevention of insulin resistance, enhancement of insulin sensitivity, prevention and treatment of diabetes, improvement of glucose metabolism, reduction of low density lipoprotein, reduction of cholesterol, reduction of obesity, hypertriglyceridemia. A pharmaceutical composition for use selected from the group consisting of prevention and treatment of disease, prevention and treatment of atherosclerotic arterial disease.
(Preparation method for unsaturated fatty acids or unsaturated fatty acid derivatives)

Derivatives of unsaturated fatty acids containing two or more double bonds (eg, EPA, DPA and DHA) or unsaturated fatty acids containing two or more double bonds, which are the raw materials of the method of the present invention, are, for example, crude fish oil. It is possible to produce a raw material prepared from an ester by treating it with the enzyme of the present invention.

Ethyl esterification methods for fractions containing fatty acids are well known. For example, the fatty acid in the fractionated glyceride fraction is ethyl esterified by an acid catalyst or an alkali catalyst or an enzyme (lipase) in the presence of ethyl alcohol. Preferably, the fatty acids in the glycerin fatty acid ester contained in the fractionated glyceride fraction are ethyl esterified by an alkali catalytic method or an enzymatic method. The fatty acids in the fractionated free fatty acid fraction are ethyl esterified by an acid catalyst or an enzyme (lipase) in the presence of ethyl alcohol. When an enzyme is used, the amount of ethyl alcohol to be added is preferably an amount that does not inactivate the lipoxygenase enzyme, preferably 4 molar equivalents or less, more preferably 2 molar equivalents or less with respect to the glyceride or free fatty acid. However, it is not limited to these.

Highly unsaturated fatty acids such as EPA and DHA in fats and oils containing ethyl esterified ω3 fatty acids can be used by the urea addition method (Japanese Patent Laid-Open No. 2007-89522, JP-A-2009-504588) and the silver nitrate complex method (Japanese Patent Laid-Open No. 2007-504588). Open 2010-64974, JP-A-7-242895), Vacuum precision distillation method including vacuum thin film distillation method (Japanese Patent Laid-Open No. 11-209786), liquid chromatography (hereinafter referred to as HPLC) and pseudo-mobile phase chromatography. Purification is performed with relatively high purity by one or more combination methods such as a chromatographic method such as a method (Japanese Patent Laid-Open No. 11-209786). You may also use SMB chromatography.

The present invention also provides methods of treating and / or preventing a disease or disorder that can be treated and / or prevented by administration of an effective amount of a therapeutic agent to a subject. Therapeutic agent means the composition of the invention in combination with a pharmaceutically acceptable carrier type (eg, sterile carrier).

Hereinafter, the present invention will be described in detail with reference to Examples and the like, but the present invention is not limited thereto.
(Example 1)
Commercially available dried soybeans (50 g) were immersed in water (150 mL) at 7 ° C. for 10 hours to remove the immersion water. In order to inactivate peroxidase in soybeans that decompose hydroperoxide, hot water (400 mL) preheated to 70 ° C. was added thereto, and the mixture was allowed to stand at 70 ° C. for 5 minutes. Subsequently, docosahexaenoic acid (0.5 g) was added thereto, and the mixture was pulverized with a mixer for 2 minutes. Those at this stage are called Namago. This is reheated to 70 ° C. for 10 minutes, and the filtrate filtered with a cloth is soymilk. In order to make the oxidation reaction by soybean lipoxygenase in the raw gourd efficient, the raw gourd was transferred to a 500 mL beaker and vigorously stirred with a magnetic stirrer for 3.5 hours. The optimum values for the amount of soybean, the amount of soaking water, the soaking temperature, the soaking time, the peroxidase deactivation temperature, the deactivation time, the amount of docosahexaenoic acid, the amount of eicosapentaenoic acid or linoleic acid, and the crushing time are described below. Values other than are also possible.

(Example 2)
It was proved by the following experiment that the metabolite produced in the raw gou or soymilk produced by the present invention was 17S-HDHA. The metabolite components contained in the soymilk and tofu produced in the present application are extracted at the stage of raw wool, separated by thin layer chromatography, isolated and purified by medium pressure chromatographic normal phase, and the structure and purity are confirmed by HPLC. , ESI MASS and ¹ H NMR. As a result, it was confirmed that the metabolite was 17S-HDHA.

Prior to the preparation of soymilk, the following experiment was first carried out to confirm the production of 17S-HDHA in raw Kure, which is the pre-stage of soymilk. Soybeans (71 g) were immersed in water (213 mL) overnight (7 ° C.), water (160 mL) was further added, and the mixture was pulverized with a mixer (4 min). This was transferred to a 1 L-nas flask with 200 mL of water and heated to around 65 ° C. with vigorous stirring to inactivate hydroperoxide lyase. DHA fatty acid (97%, 1 mL) was added thereto, and the mixture was vigorously stirred at 63 to 65 ° C. for 2.5 hours.

Of these, 100 mL was separated and an attempt was made to extract with ethyl acetate, but the mixture was not separated. Another 100 mL was made slightly acidic with 1N-HCl at around pH 4-5 and separated into two layers of solid liquid by centrifugation. The extracted solid residue was extracted with ethyl acetate. Ethyl acetate extract was obtained by centrifugation. TLC analysis of this extract (hexane / ether / acetic acid 70: 30: 1) focused on spots near Rf = 0.35, and this fraction was fractionated and purified by medium-pressure chromatographic normal phase. This fraction gave a single spot at Rf = 0.16 (Hexane / Ether / Acetic Acid, 70:30: 1) by normal phase thin layer chromatography. (HPLC: Inertsil ODS-3; 4.6x250 mm, methanol / water = 9: 1,10 mL / min.Rt = 8.13 min). Was carried out for this fraction Mass spectrometry (ESI MASS), signals appear in the m / z 343.23, this value is the calculated value m / z 343.24 for the _{17S-HDHA (C 22 H 31} O 3) It matched. Furthermore, the fragment ion-cleaved on the carboxyl-terminal side of the carbon to which the hydroxyl group is bonded is m / z 245.2, which matches the calculated value m / z 245, so that the hydroxyl group is bonded to the 17-position of DHA. The thing was proved.

(Example 3)
According to Example 1, it was confirmed that 17S-HDHA was produced from DHA fatty acid (1.0 g) by lipoxygenase, which is relatively heat-stable by heat-inactivating peroxidase during the preparation of raw wool. To confirm, the extraction method of 17S-HDHA was improved.

After soybeans (50 g) were immersed in 150 mL of water, the soaked water was discarded, soybeans were added to water heated to 70 ° C. (300 mL), heated to 70 ° C., and left for 5 minutes. DHA fatty acid (70% purity, 1.0 g) was added thereto, and the mixture was pulverized with a mixer for 2 minutes. This was heated at 70 ° C. for 10 minutes. The mixture was filtered through a cloth to obtain a filtrate (soy milk, 351 g). In order to measure the amount of 17S-HDHA contained in the soymilk obtained here, 70 g of 351 g was separated and lyophilized into powder. This powder (5.0 g) is suspended and stirred in ethanol (100 mL), the filtrate obtained by filtration is concentrated, and then placed in a 10 mL volumetric flask to make 10 mL. Analyzed in. Approximately from the comparison of spot intensities, it was shown that 10.8 mg of 17S-HDHA was contained in 351 g of soymilk.
(Experimental Example 4)
After soybeans (50 g) were immersed in 150 mL of water, the soaked water was discarded, soybeans were added to water heated to 70 ° C. (300 mL), heated to 70 ° C., and left for 5 minutes. After removing the hot water, DHA fatty acid (50% purity, 1.09 g) was added to soybean (50 g), and the mixture was pulverized with a mixer for 2 minutes. This was heated at 70 ° C. for 10 minutes. The obtained filtrate (soy milk, 351 g) was obtained by filtering with a cloth. In order to measure the amount of 17S-HDHA contained in the soymilk obtained here, 70 g of 351 g was separated and lyophilized into powder. The amount of 17S-HDHA contained in this powder was quantified by HPLC-HS MASS. The amount of 17S-HDHA in 100 g of soymilk was 1.7 mg.
(Example 5)
The same experiment as in Example 3 was performed using DHA fatty acid (97% purity, 1.9 g). As a result, the amount of 17S-HDHA in 100 g of soymilk was 11.9 mg.
(Example 6)
An experiment similar to Example 4 was performed using triglyceride (2.0 mL) containing 70% DHA. As a result, 17S-HDHA was not detected by HPLC MS in the powdered soymilk sample obtained from this sample. Therefore, this TG sample was alkaline hydrolyzed to make it slightly acidic, and then the extracted ether solution contained 17S-HDHA, which was 0.42 mg / 100 g soymilk in terms of soymilk. It was clarified that the soymilk obtained by heat-treating the soybeans in this way also acts on triglycerides that bind unsaturated fatty acids and gives metabolites.

Examples 1-6 relate to the production of soymilk containing the unsaturated fatty acid metabolite 17S-HDHA, but the following examples show that the method of the present application is also effective in the production of pharmaceutical compositions. .. In order to investigate the possibility, we tried to use soymilk as an enzyme reagent instead of lipoxygenase as follows.

(Example 7)
Boric acid buffer (pH 9,200 mL) was added to the soymilk (100 g, without DHA) obtained by the method of Example 3, DHA fatty acid (97% purity, 1 g) was added thereto, and oxygen gas was added while stirring. Was blown in for 6 hours. The reaction solution is adjusted to pH 6 to 6.5 with dilute hydrochloric acid (0.1N), extracted with ethyl acetate (3 times) in a liquid separation funnel, washed with water (3 times), dried with anhydrous magnesium sulfate, and filtered. As a result of concentration and purification in the reverse phase of medium pressure chromatography, 17S-HDHA could be obtained in a yield of 20%.

(Example 8)
In Example 3, during the preparation of raw wool, peroxidase was heat-inactivated, while lipoxygenase, which is relatively heat-stable, was suppressed in heat-inactivation, and the resulting enzyme mixture was used to obtain DHA fatty acid (1.0 g). ), And the amount thereof was determined. Using this quantification method, it is possible to determine preferable conditions for heat-treating a composition containing lipoxygenase.
As described in Example 3, soybeans (50 g) are immersed in 150 mL of water, the immersion water is discarded, and soybeans are added to water (300 mL) heated to various temperatures (for example, 70 ° C.) to heat them. .. The heating time can be changed as appropriate. Then, it is quickly cooled to a predetermined temperature in the range of room temperature to 37 ° C. DHA fatty acid (70% purity, 0.5 g) was added to the enzyme mixture thus obtained, and the mixture was ground with a mixer for 2 minutes. This was heated at 70-75 ° C. for 10 minutes. The mixture was filtered through a cloth, and the filtrate (soy milk, 351 g) was vigorously stirred for 4 hours. In order to measure the amount of 17S-HDHA contained in the soymilk obtained here, 70 g of 351 g was separated and lyophilized into powder. This powder (5.0 g) is suspended and stirred in ethanol (100 mL), the filtrate obtained by filtration is concentrated, and then placed in a 10 mL volumetric flask to make 10 mL. Analyzed in. The amount of 17S-HDHA is estimated from the comparison of spot intensities. These methods include enzyme mixtures (eg, soybean flour, bean soak, raw sardines), and soymilk that have been subjected to a variety of different heating conditions (eg, different temperatures and different heating times). ) Is evaluated. It is possible to determine the optimum heating conditions by this method.
(Example 9)
Up to Example 8, the synthesis of 17S-HDHA using DHA fatty acid as a substrate has been mainly described, but in Example 9, the result of using EPA fatty acid will be described. In the same manner as in Example 3, soybeans (produced in Hokkaido, 100 g) were immersed in water (300 mL) for 17.5 hours (7 ° C.) to remove most of the immersion water. The soybeans (222.5 g) and hot water (70 ° C., 400 mL) were placed in a beaker and heated for 5 minutes. After draining the hot water, the soybeans were divided into two (A: 111 g, B: 111 g).
(Example 10)
Synthesis of 15-hydroxyeicosapentaenoic acid (15-Hydroxy-eicosapentaenoic acid, 15-HEPE), an EPA metabolite by soymilk.
A (111 g) and hot water (70 ° C., about 200 mL) were added to a mixer, EPA fatty acid (1.0 mL) was added thereto, and the mixture was pulverized for 2 minutes. When the temperature dropped to some extent, it was filtered with a cloth to obtain soymilk (154 g). This was stirred at room temperature for 2.5 hours. This sample was lyophilized to give a powder (20.4 g). 5 g of this powder was extracted with ethanol (100 mL) for 29 hours. After refrigerating, the solvent was removed with an evaporator (40 ° C.) to obtain 1.08 g of a sample. This was dissolved in 1 ml of ethyl acetate, and it was confirmed that 15-HEPE was produced by TLC (hexane: ether: acetic acid = 70: 30: 1). The Rf value was 0.21. It was shown that 154 g of soymilk contained 4 mg of 15-HEPE in terms of spot intensity.
(Example 11)
Synthesis of 15-HEPE with soybean whey.
B (111 g) and hot water (70 ° C., about 200 mL) were added to the mixer, and the mixture was pulverized for 2 minutes. When the temperature dropped to some extent, it was filtered with a cloth to obtain soymilk (200 g). This was heated to 65 ° C., and 10 mL of bittern liquid (10 mL, a mixture of commercially available bittern liquid (12.5 mL) and hot water (50 mL)) was added. The tofu-like solid was filtered through a cloth, EPA fatty acid (0.3 mL) was added to the filtrate (soybean whey, 48 g), and the mixture was stirred at room temperature for 16 hours. The powder (2.6 g) was obtained by lyophilization. Of this, 5 g was extracted with ethanol (100 mL) for 27 hours. The solvent was removed with an evaporator (40 ° C.) to obtain 0.77 g of a sample. This was dissolved in ethyl acetate, and it was confirmed that 15-HEPE was produced by TLC (hexane: ether: acetic acid = 70: 30: 1). The Rf value at this time was 0.21. It was shown that 48 g of soymilk whey contained 4 mg of 15-HEPE in terms of spot intensity.
(Example 12)
Synthesis of 13-hydroxyoctadecadienic acid (13-HODA), a metabolite of linoleic acid by soymilk.
Hot water (70 ° C., about 400 mL) was added to the soybeans (111 g) obtained in the same manner as in Example 10, linoleic acid (1.0 mL) was added thereto, and the mixture was pulverized for 2 minutes. When the temperature dropped to some extent, it was filtered with a cloth to obtain soymilk (334.8 g). This was stirred at room temperature for 3 hours. 30 g was collected from this sample, 100 ml of ethanol was added, and the mixture was stirred at room temperature for 2 hours. After filtering to remove the solids, the solvent was removed with an evaporator to obtain an 800 mg sample. Further, 20 ml of ethyl acetate was added, and the ethyl acetate layer was recovered. The solid content was removed by filtration again, and the solvent was removed by an evaporator to obtain a 350 mg sample. This was dissolved in methanol and purified by medium pressure chromatography. As a result, 110 mg was recovered. This was dissolved in 10 ml of methanol and analyzed by TLC (hexane: ether: acetic acid = 50: 50: 1), and it was confirmed that a linoleic acid metabolite was produced. The Rf value was 0.32. It was shown that 1.23 mg of linoleic acid metabolite 13-HODA was contained in 334.8 g of soymilk in terms of spot intensity.
(Example 13)
Synthesis of 13-hydroxyoctadecadienoic acid (13-HODA), a metabolite of linoleic acid by soybean whey.
Soybeans (111 g) obtained in the same manner as in Example 10 and hot water (70 ° C., about 400 mL) were added and pulverized for 2 minutes. When the temperature dropped to some extent, it was filtered with a cloth to obtain soymilk. This was heated at 70 ° C. for an additional 10 minutes. Subsequently, a bittern solution (10 mL, a mixture of a commercially available bittern solution (12.5 mL) and hot water (50 mL)) was added, and the mixture was allowed to stand for 20 minutes. The tofu-like solid was filtered through a cloth, linoleic acid (0.5 mL) was added to the filtrate (soybean whey, 72.4 g), and the mixture was stirred at room temperature for 16 hours. The linoleic acid metabolite was extracted twice with ethyl acetate (70 mL) from this reaction solution. It was washed with water (twice) and dried over sodium sulfate. After removing sodium sulfate by filtration, the filtrate was concentrated to obtain 0.5 g of a residue. Methanol (5.0 mL) was added thereto, and it was confirmed that a linoleic acid metabolite was produced by TLC (hexane: ether: acetic acid = 50: 50: 1). The Rf value at this time was 0.27. It was shown that 0.22 mg of linoleic acid metabolite 13-HODA was contained in 72.4 g of soymilk whey in terms of spot intensity.

As described above, the present invention has been illustrated using the preferred embodiment of the present invention, but the present invention should not be construed as being limited to this embodiment. It is understood that the invention should be construed only by the claims. It will be understood by those skilled in the art that from the description of specific preferred embodiments of the present invention, an equivalent range can be implemented based on the description of the present invention and common general technical knowledge. The patents, patent applications and documents cited herein are to be incorporated by reference in their content as they are specifically described herein. Understood.

The present invention is novel and has excellent advantages over the prior art, metabolism of a substance consisting of an unsaturated fatty acid containing two or more double bonds or a derivative of an unsaturated fatty acid containing two or more double bonds. Provided are a method for producing an edible composition and a pharmaceutical composition containing a substance.

Claims (28)

A method for producing an edible composition containing a metabolite of a substance consisting of an unsaturated fatty acid containing two or more double bonds or a derivative of an unsaturated fatty acid containing two or more double bonds.
(A) A step of providing a first composition containing a substance selected from the group consisting of unsaturated fatty acids containing two or more double bonds and derivatives of unsaturated fatty acids containing two or more double bonds;
(B) A step of providing a heat-treated second composition, wherein the second composition contains lipoxygenase; and (c) the first composition and the first. A step of mixing the two compositions and reacting the lipoxygenase with the substance contained in the first composition.
Including, methods. The method according to claim 1, wherein the heat treatment in the step (b) is performed at 60 ° C. to 90 ° C. The method according to claim 1, wherein the second composition is a bean-derived lipoxygenase-containing composition. The method of claim 1, wherein the second composition comprises a substance selected from the group consisting of soybeans, soymilk, soybean flour, soybean flour, whey, and soybean soaked liquid. The method according to claim 1, wherein the second composition is a composition containing a substance derived from soybean. The method of claim 1, wherein the unsaturated fatty acid is selected from the group consisting of omega-3 fatty acids and omega-6 fatty acids. The unsaturated fatty acid is selected from the group consisting of DHA, EPA, DPA (n-3), DPA (n-6), linoleic acid, α-linolenic acid, γ-linolenic acid, stearidonic acid, and arachidonic acid. , The method according to claim 1. The method according to claim 6, wherein the omega 3 fatty acid is EPA or DHA. The method according to claim 6, wherein the omega-6 fatty acid is linoleic acid. The method of claim 1, wherein the second composition comprises soybean whey. The method according to claim 1, wherein the second composition comprises soybean raw gou (soybean). The method according to claim 1, wherein the metabolite of the unsaturated fatty acid or the metabolite of the derivative of the unsaturated fatty acid is 17S-HDHA, 17S-HDHA ethyl ester, 17S-HDHA-bound triglyceride, or 17S-HDHA-bound phosphorus. Lipids, 15-HEPE, 15-HEPE ethyl ester, 15-HEPE-linked triglyceride, 15-HEPE-linked phospholipid, 17-HDPA, 17-HDPA ethyl ester, 17-HDPA-linked triglyceride, 17-HDPA-linked phospholipid, 13- Hydroxylinolic acid, 13-hydroxylinolic acid ethyl ester, 13-hydroxylinolic acid bound triglyceride, 13-hydroxylinolic acid bound phospholipid, 13-hydroxy-α-linolenic acid, 13-hydroxy-α-linolenic acid ethyl ester, 13 -Hydroxy-α-linolenic acid bound triglyceride, 13-hydroxy-α-linolenic acid bound phospholipid, 13-hydroxy-γ-linolenic acid, 13-hydroxy-γ-linolenic acid ethyl ester, 13-hydroxy-γ-linolenic acid Bound triglyceride, 13-hydroxy-γ-linolenic acid-bound phospholipid, 13-hydroxystearidonic acid, 13-hydroxystearidonic acid ethyl ester, 13-hydroxystearidonic acid-bound triglyceride, 13-hydroxystearidonic acid-bound phospholipid, 15-Hydroxyarachidonic acid, 15-hydroxyarachidonic acid ethyl ester, 15-hydroxyarachidonic acid-bound triglyceride, 15-hydroxyarachidonic acid-bound phospholipid, 13-hydroxyoctadecadienoic acid (13-HODA), 13-hydroxyoctadecadiene A method selected from the group consisting of acid (13-HODA) ethyl ester, 13-hydroxyoctadecadienoic acid (13-HODA) -linked triglyceride, and 13-hydroxyoctadecadienoic acid (13-HODA) -linked phospholipids. .. A method for producing an edible composition containing a metabolite of a substance consisting of an unsaturated fatty acid containing two or more double bonds or a derivative of an unsaturated fatty acid containing two or more double bonds.
(A) A step of providing a composition containing a lipoxygenase and a substance selected from the group consisting of unsaturated fatty acids containing two or more double bonds and derivatives of unsaturated fatty acids containing two or more double bonds. And (b) the step of heat-treating the composition,
Including, methods. The method according to claim 13, wherein the heat treatment in the step (b) is performed at 60 ° C. to 90 ° C. 13. The method of claim 13, wherein the lipoxygenase is a bean-derived lipoxygenase. 13. The method of claim 13, wherein the composition comprises a substance selected from the group consisting of soybeans, soymilk, soybean flour, soybean flour, whey, and soybean soaking liquor. The method according to claim 13, wherein the composition is a composition containing a substance derived from soybean. 13. The method of claim 13, wherein the unsaturated fatty acid is selected from the group consisting of omega-3 fatty acids and omega-6 fatty acids. The unsaturated fatty acid is selected from the group consisting of DHA, EPA, DPA (n-3), DPA (n-6), linoleic acid, α-linolenic acid, γ-linolenic acid, stearidonic acid, and arachidonic acid. , The method according to claim 13. The method according to claim 18, wherein the omega 3 fatty acid is EPA or DHA. The method of claim 18, wherein the omega-6 fatty acid is linoleic acid. 13. The method of claim 13, wherein the composition comprises soybean whey. 13. The method of claim 13, wherein the composition comprises soybean raw gou. The method according to claim 13, wherein the metabolite of the unsaturated fatty acid or the metabolite of the derivative of the unsaturated fatty acid is 17S-HDHA, 17S-HDHA ethyl ester, 17S-HDHA-bound triglyceride, or 17S-HDHA-bound phosphorus. Lipids, 15-HEPE, 15-HEPE ethyl ester, 15-HEPE-linked triglyceride, 15-HEPE-linked phospholipid, 17-HDPA, 17-HDPA ethyl ester, 17-HDPA-linked triglyceride, 17-HDPA-linked phospholipid, 13- Hydroxylinolic acid, 13-hydroxylinolic acid ethyl ester, 13-hydroxylinolic acid bound triglyceride, 13-hydroxylinolic acid bound phospholipid, 13-hydroxy-α-linolenic acid, 13-hydroxy-α-linolenic acid ethyl ester, 13 -Hydroxy-α-linolenic acid bound triglyceride, 13-hydroxy-α-linolenic acid bound phospholipid, 13-hydroxy-γ-linolenic acid, 13-hydroxy-γ-linolenic acid ethyl ester, 13-hydroxy-γ-linolenic acid Bound triglyceride, 13-hydroxy-γ-linolenic acid-bound phospholipid, 13-hydroxystearidonic acid, 13-hydroxystearidonic acid ethyl ester, 13-hydroxystearidonic acid-bound triglyceride, 13-hydroxystearidonic acid-bound phospholipid, 15-Hydroxyarachidonic acid, 15-hydroxyarachidonic acid ethyl ester, 15-hydroxyarachidonic acid-bound triglyceride, 15-hydroxyarachidonic acid-bound phospholipid, 13-hydroxyoctadecadienoic acid (13-HODA), 13-hydroxyoctadecadiene A method selected from the group consisting of acid (13-HODA) ethyl ester, 13-hydroxyoctadecadienoic acid (13-HODA) -linked triglyceride, and 13-hydroxyoctadecadienoic acid (13-HODA) -linked phospholipids. .. An edible composition comprising a metabolite of the unsaturated fatty acid or a metabolite of a derivative of the unsaturated fatty acid produced by the method according to claim 1 or 13. A method for producing a pharmaceutical composition containing a metabolite of a substance consisting of an unsaturated fatty acid containing two or more double bonds or a derivative of an unsaturated fatty acid containing two or more double bonds.
(A) A step of providing a first composition containing a substance selected from the group consisting of unsaturated fatty acids containing two or more double bonds and derivatives of unsaturated fatty acids containing two or more double bonds;
(B) A step of providing a heat-treated second composition, wherein the second composition contains lipoxygenase; and (c) the first composition and the first. A step of mixing the two compositions and reacting the lipoxygenase with the substance contained in the first composition.
Including, methods. A method for producing a pharmaceutical composition containing a metabolite of a substance consisting of an unsaturated fatty acid containing two or more double bonds or a derivative of an unsaturated fatty acid containing two or more double bonds.
(A) A step of providing a composition containing a lipoxygenase and a substance selected from the group consisting of unsaturated fatty acids containing two or more double bonds and derivatives of unsaturated fatty acids containing two or more double bonds. And (b) the step of heat-treating the composition,
Including, methods. A pharmaceutical composition comprising a metabolite of the unsaturated fatty acid or a metabolite of a derivative of the unsaturated fatty acid produced by the method according to claim 26 or 27.