JP2002188096A - New glyceride, method for producing the same and its use - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new glyceride having an inhibitory function on fat storage, promotion function on reduction of deposit fats and amelioration function on carbohydrate metabolism abnormality, capable of being expected to have preventing and ameliorating effect on life-style related diseases caused by obesity. SOLUTION: This glyceride contains one or two conjugated highly unsaturated fatty acids having a conjugated triene structure or a conjugated highly unsaturated fatty acid having a conjugated triene structure and at least one kind selected from the group consisting of a 2-5C short-chain fatty acid, a 6-12 medium-chain fatty acid, a 20-22C long-chain saturated fatty acid, an 18C fatty acid having a conjugated diene structure, an 18-22C highly unsaturated fatty acid having 3-6 degree of unsaturation and a group represented by -PO3H-B [B is choline, serine, ethanolamine or inositol group].

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a novel glyceride containing a conjugated polyunsaturated fatty acid having a conjugated triene structure, a method for producing the same, and a use thereof.

[0002]

2. Description of the Related Art Glycerides comprising conjugated polyunsaturated fatty acids having a conjugated triene structure are hardly found in general animal / vegetable oils and fats, and are slightly contained in pomegranate, bitter melon,
It is only found in the seeds of a limited number of plants, such as calendula, catalpa, balsam apple, snake guard, cherries and abragiri. As conjugated polyunsaturated fatty acids contained in glycerides obtained from these plant seeds, punicic acid (18: 3,9c,
11t, 13c), calendic acid
(18: 3, 8t, 10t, 12c), jarcalic acid (18: 3, 8c, 10t, 12c)
c), α-eleostealic acid
(18: 3,9c, 11t, 13t), β-eleostealic acid (18: 3,9t,
11t, 13t), catalpic acid
(18: 3, 9t, 11t, 13c), camulolenic acid (18OH, 9c, 11t, 13c)
t) and the like are recognized. For example, in the case of glycerides derived from pomegranate seeds, punicic acid is contained in about 70% (weight%, the same applies hereinafter), palmitic acid, stearic acid, oleic acid,
It is said to contain about 30% of linolenic acid (MIEl-Shaaraway and A. Nahapetian, FETTE. SE
IFEN. ANSTRICHMITTEL, 85, 123, 1993).

[0003] On the other hand, it is known that short-chain or medium-chain fatty acids have a lower energy value than long-chain fatty acids, and that long-chain saturated fatty acids having 20 to 22 carbon atoms have poor digestive and absorptive properties. Known fatty acids having a conjugated diene structure and polyunsaturated fatty acids represented by docosahexaenoic acid are known to have a function of suppressing fat accumulation and a function of accelerating the reduction of accumulated fat, but all have problems such as strength of function and flavor. Is left. In addition, it has been reported that diglyceride exerts a function of suppressing fat accumulation, but a large amount of ingestion is required to exert the function, which is not satisfactory.

[0004]

DISCLOSURE OF THE INVENTION An object of the present invention is to provide a novel glyceride which has a function of suppressing fat accumulation and a function of accelerating reduction of accumulated fat and which can be expected to have an effect of preventing and improving lifestyle-related diseases based on obesity, and a method for producing the same. And to provide its use.

[0005]

Means for Solving the Problems Heretofore, little research has been conducted on the physiological function of glycerides composed of conjugated polyunsaturated fatty acids having a conjugated triene structure.
The present inventors have proposed that a conjugated polyunsaturated fatty acid having a conjugated triene structure can be used as a peroxisome proliferator-activated receptor (PP
AR: peroxisome proliferator-activated receptor)
We have found that it is an agonist and that its glyceride exerts a remarkable function of suppressing fat accumulation and accelerating the function of accumulating fat reduction, and applied for a patent (JP-A-2000-355538).
Thereafter, a glyceride containing one or two conjugated polyunsaturated fatty acids having a conjugated triene structure in the molecule, in order to obtain a novel glyceride having a higher fat accumulation suppressing function, a stored fat reduction promoting function or a new function, Or, together with a conjugated polyunsaturated fatty acid having a conjugated triene structure in the molecule, a short-chain fatty acid, a medium-chain fatty acid, a long-chain fatty acid, a fatty acid having a conjugated diene structure, a non-conjugated polyunsaturated fatty acid or -PO ₃ H-B is used. A novel glyceride containing a group [B represents choline, serine, ethanolamine or inositol group] was produced and its properties were examined. As a result, the novel glyceride was found to have an excellent function of suppressing fat accumulation and reducing accumulated fat. It has an accelerating function and a function of improving carbohydrate metabolism abnormalities, and has also been found to have excellent properties in terms of physical properties. A result of repeated 討, which resulted in the completion of the present invention.

That is, a first aspect of the present invention is to contain one or two conjugated polyunsaturated fatty acids having a conjugated triene structure or a conjugated polyunsaturated fatty acid having a conjugated triene structure and a conjugated polyunsaturated fatty acid having 2 to 5 carbon atoms. Short-chain fatty acids of 6 to 12 carbon atoms
Medium-chain fatty acids, long-chain saturated fatty acids having 20 to 22 carbon atoms, fatty acids having a conjugated diene structure having 18 carbon atoms, 18 carbon atoms
To 22 in degree of unsaturation (carbon - carbon unsaturated bond) 3-6 polyunsaturated fatty acids and -PO ₃ groups represented by H-B [B
Represents any one of a choline, serine, ethanolamine and inositol group], wherein the glyceride contains at least one selected from the group consisting of:

[0007] As a preferred embodiment, (1) TA
T, TTA, ATA, THT, TAP, TTP or T
HP [T is a conjugated polyunsaturated fatty acid residue having a conjugated triene structure, A is a short-chain fatty acid residue having 2 to 5 carbon atoms, a medium-chain fatty acid residue having 6 to 12 carbon atoms, and a long chain having 20 to 22 carbon atoms. Any one selected from a chain saturated fatty acid residue, a fatty acid residue having a conjugated diene structure having 18 carbon atoms, and a highly unsaturated fatty acid residue having 18 to 22 carbon atoms and an unsaturation degree of 3 to 6, H is a hydrogen atom, P
It is (choline of B, serine, either indicating the ethanolamine or inositol residue) -PO ₃ H-B glycerides described above represented by, a conjugated polyunsaturated fatty acids with (2) a conjugated triene structure Glyceride as described above, which is at least one selected from the group consisting of punicic acid, α-eleostearic acid, β-eleostearic acid, jalcalic acid, calendic acid and catalpinic acid; (3) carbon number 18 To 22 and a polyunsaturated fatty acid having an unsaturation degree of 3 to 6 is eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, γ-linolenic acid and α-
The glyceride according to the above, which is at least one selected from the group consisting of linolenic acid, (4) a function of suppressing fat accumulation,
The glyceride described above, which has at least one of a function of accelerating the reduction of accumulated fat and a function of improving abnormal carbohydrate metabolism.

A second aspect of the present invention is to provide a conjugated polyunsaturated fatty acid having a conjugated triene structure or an ester derivative thereof,
Short-chain fatty acids having 2 to 5 carbon atoms, medium-chain fatty acids having 6 to 12 carbon atoms, long-chain saturated fatty acids having 20 to 22 carbon atoms, fatty acids having a conjugated diene structure having 18 carbon atoms, unsaturated having 18 to 22 carbon atoms The present invention relates to the above-mentioned method for producing glyceride, comprising subjecting at least one selected from the group consisting of polyunsaturated fatty acids having a degree of 6 to 6 and their ester derivatives and phospholipids to a transesterification reaction. In a preferred embodiment, the ester derivative of the conjugated polyunsaturated fatty acid is a pomegranate seed oil, a bitter melon seed oil, a calendula seed oil, a ginseng seed oil, a catalpa seed oil, an American cattle seed oil, a balsam apple seed oil, a snake guard seed. The method according to the above, wherein the glyceride is obtained from at least one vegetable oil selected from the group consisting of oil, oilseed seed oil, and cherry seed oil.

A third aspect of the present invention comprises at least one of the glycerides described above and has at least one of a function of suppressing fat accumulation, a function of accelerating reduction of accumulated fat, and a function of improving abnormal carbohydrate metabolism. The present invention relates to a fat composition. A fourth aspect of the present invention relates to a food comprising the above-described fat or oil composition. A fifth aspect of the present invention relates to a pharmaceutical product containing the above-described fat or oil composition. A sixth aspect of the present invention relates to a feed containing the above-described fat or oil composition.

Heretofore, glycerides containing one or two conjugated polyunsaturated fatty acids having a conjugated triene structure, and conjugated polyunsaturated fatty acids having a conjugated triene structure, as well as short-chain fatty acids having 2 to 5 carbon atoms, Carbon number 6 ~
12 medium-chain fatty acids, long-chain saturated fatty acids having 20 to 22 carbon atoms, fatty acids having a conjugated diene structure having 18 carbon atoms, highly unsaturated fatty acids having 18 to 22 carbon atoms and an unsaturated degree of 3 to 6, or -PO ₃ H A group represented by -B [B is choline, serine,
Which shows at least one selected from the group consisting of ethanolamine and inositol groups].

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail.

The glyceride of the present invention contains one or two conjugated polyunsaturated fatty acids having a conjugated triene structure (hereinafter referred to as “conjugated triene polyunsaturated fatty acids”), or conjugated triene polyunsaturated fatty acids. And a short-chain fatty acid having 2 to 5 carbon atoms, a medium-chain fatty acid having 6 to 12 carbon atoms, a long-chain saturated fatty acid having 20 to 22 carbon atoms, a fatty acid having a conjugated diene structure having 18 carbon atoms, and having 18 to 22 carbon atoms. contains at least one [the B shown choline, serine, ethanolamine or inositol radical group represented by polyunsaturated fatty acids degree of unsaturation 3-6 or -PO ₃ H-B is selected from the group consisting of Glyceride. The form of glyceride may be any of triglyceride, diglyceride and monoglyceride, and it is sufficient that the glyceride has at least one conjugated triene highly unsaturated fatty acid residue.

Further, in the glyceride of the present invention, a conjugated triene highly unsaturated fatty acid, a short chain fatty acid having 2 to 5 carbon atoms, a medium chain fatty acid having 6 to 12 carbon atoms, a long chain saturated fatty acid having 20 to 22 carbon atoms, fatty acids having a conjugated diene structure having 18 carbon atoms, groups [B represented by polyunsaturated fatty acids or -PO ₃ H-B degrees of unsaturation 3-6 at 18-22 carbon atoms choline, serine, ethanolamine or Which represents an inositol group] is not particularly limited and may be appropriately selected depending on the intended purpose.
ATA, THT, TAP, TTP or THP [T: conjugated triene highly unsaturated fatty acid residue, A: short-chain fatty acid residue having 2 to 5 carbon atoms, medium-chain fatty acid residue having 6 to 12 carbon atoms, 20 carbon atoms Any one selected from a long-chain saturated fatty acid residue having a conjugated diene structure having 18 carbon atoms, a highly unsaturated fatty acid residue having 18 to 22 carbon atoms and an unsaturated degree of 3 to 6, H: A glyceride represented by a hydrogen atom and a group represented by P: —PO ₃ H—B (B represents a choline, serine, ethanolamine or inositol residue)] is preferable, and among these glycerides, fat accumulation is suppressed. Excellent function, function to promote accumulated fat reduction, function to improve abnormal carbohydrate metabolism,
TAT, ATA, THT, and TAP are more preferred.

The conjugated triene highly unsaturated fatty acid in the glyceride of the present invention is not particularly limited as long as it is a fatty acid having a conjugated triene structure.
α-eleostearic acid, β-eleostearic acid, jalcalic acid, calendic acid, catalpic acid, camlorenic acid and the like. Among them, punicic acid, α-eleostearic acid, β-eleostearic acid, jalcalic acid, calendic acid, and catalpic acid, which are easily available from the raw materials, are preferable, and they have stability, economy, a function of suppressing fat accumulation, and accumulation. Punicic acid and α-eleostearic acid are most preferred from the viewpoint of the function of promoting fat reduction. Conjugated triene polyunsaturated fatty acids and ester derivatives thereof may be obtained by chemically conjugate isomerization of polyunsaturated fatty acids, but plants containing conjugated triene polyunsaturated fatty acid glycerides contained in natural plant seeds It is preferred to use seed oil. Preferred plant seed oils include pomegranate seed oil, bitter melon seed oil, calendula seed oil, rosewood seed oil, catalpa seed oil, American catalpa seed oil,
Examples include balsam apple seed oil, snake guard seed oil, abragiri seed oil or cherry seed oil. Of these, easily available pomegranate seed oil and bitter melon seed oil are more preferred. These fats and oils may be used directly as a raw material for producing the glyceride of the present invention, or may be used after being hydrolyzed with an alkali or the like, and then used as a fatty acid or further derived into a methyl ester or the like by fractionated solvent extraction or column chromatography. Good, but it is convenient to use the oils and fats extracted from plant seeds as they are.

The short-chain fatty acid in the glyceride of the present invention is not particularly limited as long as it has 2 to 5 carbon atoms, and examples thereof include acetic acid, propionic acid, butyric acid, and valeric acid.

The medium-chain fatty acid in the glyceride of the present invention is not particularly limited as long as it has 6 to 12 carbon atoms.
For example, caproic acid, heptyl acid, caprylic acid, capric acid, lauric acid may be mentioned. For these, commercially available reagents may be used, or fats and oils containing these fatty acids at a high content may be used. For example, palm kernel oil and coconut oil containing about 50% lauric acid may be mentioned.

The long-chain saturated fatty acid in the glyceride of the present invention is not particularly limited as long as it has 20 to 22 carbon atoms, and examples thereof include arachidic acid and behenic acid. These may use commercially available reagents.

The fatty acid having 18 carbon atoms and a conjugated diene structure in the glyceride of the present invention is not particularly limited as long as it is octadecadienoic acid, and the position and configuration of the conjugated diene are (9c, 11t), (9c , 11c), (9t, 1
1c), (9t, 11t), (10c, 12c), (10
c, 12t), (10t, 12c) and (10t, 12
Although it is appropriately selected from t), (9c, 11t) or (9t, 11c) having a strong function of suppressing fat accumulation is preferable.

The glyceride of the present invention has 18 to 18 carbon atoms.
Highly unsaturated fatty acids having a degree of unsaturation of 3 to 6 at 22 are not limited by the position of the unsaturated bond and its configuration,
It refers to those other than the conjugated triene highly unsaturated fatty acids.
Specific examples include α-linolenic acid, γ-linolenic acid, dihomo-α-linolenic acid, dihomo-γ-linolenic acid, arachidonic acid, eicosatetraenoic acid, eicosapentaenoic acid, docosatetraenoic acid, and docosa. Although pentaenoic acid and docosahexaenoic acid can be mentioned, eicosapentaenoic acid and docosahexaenoic acid are preferred from the viewpoint of the function of suppressing fat accumulation. In addition, these fatty acids may be hydroxylated, epoxidized or hydroxyepoxidized. These fatty acids and their ester derivatives can be purchased and used as reagents, or oils and fats containing these fatty acids at a high level may be used. For example, oils and oils containing high levels of α-linolenic acid include perilla oil and linseed oil, and oils and oils containing high levels of γ-linolenic acid include evening primrose seed oil, eicosapentaenoic acid or docosahexaenoic acid. Examples of fats and oils containing a high level of include fish oils such as refined tuna orbital oil.

Examples of glycerides having a group represented by -PO ₃ H-B [B represents a choline, serine, ethanolamine or inositol group] include phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine and phosphatidylinositol. A reagent may be obtained and used, or soybean lecithin or egg yolk lecithin used as food may be used.

The glyceride of the present invention comprises a glyceride comprising a conjugated triene highly unsaturated fatty acid, a short chain fatty acid, a medium chain fatty acid, a long chain fatty acid, a fatty acid having a conjugated diene structure,
It can be produced by transesterification with non-conjugated highly unsaturated fatty acids and their ester derivatives or phospholipids.

In the production of the glyceride of the present invention, an ester derivative of each fatty acid can be used as a raw material. The form of the ester is not particularly limited, and includes, for example, methyl ester, ethyl ester, benzyl ester and the like, and an easy-to-use form may be used according to the purpose. Furthermore, short chain fatty acids having 2 to 5 carbon atoms, 6 to 5 carbon atoms
12 medium-chain fatty acids, long-chain saturated fatty acids having 20 to 22 carbon atoms, fatty acids having a conjugated diene structure having 18 carbon atoms, highly unsaturated fatty acids having 18 to 22 carbon atoms and a degree of unsaturation of 3 to 6, or ester derivatives thereof or a group represented by -PO ₃ H-B [B choline, serine, showing the ethanolamine or inositol group] may also be used glycerides having.

The glyceride of the present invention comprises the above conjugated triene highly unsaturated fatty acid or its ester derivative, a short-chain fatty acid having 2 to 5 carbon atoms, a medium-chain fatty acid having 6 to 12 carbon atoms,
The raw material is a long-chain saturated fatty acid having 20 to 22 carbon atoms, a fatty acid having a conjugated diene structure having 18 carbon atoms, a highly unsaturated fatty acid having 18 to 22 carbon atoms and a degree of unsaturation of 3 to 6 or an ester derivative or phospholipid thereof. It can be prepared by transesterification, hydrolysis or a combination of transesterification and hydrolysis.

The transesterification method used in the present invention is not particularly limited, and may be a chemical method or a method using an enzyme. In the case of a chemical method, for example, about 0.1 to 1% of a catalyst such as sodium methoxide is added to a mixture of glyceride composed of conjugated triene polyunsaturated fatty acid and palm kernel oil, and the mixture is heated at 80 ° C for 30 minutes under vacuum. After stirring to a certain extent, water can be added to deactivate the catalyst, followed by dehydration, decolorization, and deodorization. The transesterification reaction may be performed by a method using an enzyme. In this case, the enzyme may be in the form of powder, liquid, or immobilized, and the reaction may be performed by a batch method or a column method, but the enzyme can be used repeatedly and can be easily separated after the reaction. Enzymes are preferred.

The enzyme that can be used for transesterification or hydrolysis is not particularly limited as long as it is a lipase that hydrolyzes fats and oils, and it is preferable to use a lipase having regiospecificity as required or required. The lipase is not particularly limited to its origin, manufacturer, or preparation method, and may be a purified lipase, a crude enzyme or a culture of a microorganism producing the enzyme, or the like.
It is convenient to use a commercially available enzyme. Examples of commercially available enzymes include mold-derived lipase A “Amano” and lipase M
Examples include "Amano", lipase F "Amano", lipase AY "Amano", and Nulase F (all manufactured by Amano Pharmaceutical Co., Ltd.). When an immobilized enzyme is used, there is no restriction on the carrier and the immobilization method, and it is sufficient to use a commercially available immobilized enzyme. Examples of commercially available immobilized lipase include Lipozyme IM (manufactured by Novo Nordisk).

When the glyceride of the present invention is produced from a glyceride having a group represented by -PO ₃ H-B (B represents a choline, serine, ethanolamine or inositol group), the enzyme described above is used as an enzyme. In addition to lipases, it is preferable to use phospholipases. Phospholipase used is not subject to particular limitation so long as it is used in the transesterification of phospholipids, usually include phospholipase A ₁ or phospholipase A _2. These phospholipases are not particularly limited by their origin, manufacturer, and preparation method, and even purified lipases
The enzyme may be a crude enzyme or a culture of a microorganism producing the enzyme.

[0027] Among the glycerides of the present invention, the above-mentioned TAT,
The production of ATA is carried out, for example, by mixing and dissolving pomegranate seed oil and palm kernel oil in n-hexane, and then mixing the lipozyme IM
(Manufactured by Novo Nordisk) and added at 40 ° C for 24 hours.
After stirring for a time, the lipozyme IM can be filtered off and the solvent can be distilled off.

For example, TTA is produced by hydrolyzing pomegranate seed oil with 1,3-specific lipase.
May be prepared by subjecting a mixture of punicic acid and docosahexaenoic acid to a transesterification reaction.

The THT may be obtained by hydrolyzing a glyceride comprising a conjugated triene polyunsaturated fatty acid with a 2-specific lipase and then removing the liberated fatty acid. The glyceride may be prepared by hydrolyzing a glyceride with lipase to obtain a monoglyceride, and then transesterifying it with a conjugated triene highly unsaturated fatty acid.

TAP is prepared from the above phospholipids such as phosph
Apatidylcholine is converted to phospholipase A ₁Enzyme with activity
After hydrolyzing to obtain a 2-acyl-phospholipid,
Esthetic treatment using pomegranate seed oil and 1,3-specific lipase
Can be obtained by exchanging them.

THP can be obtained by treating the TAP obtained above with 2-specific phospholipase, and purifying the lysophospholipid from the product using a silica gel column or the like.

The ATP may, for example, phosphatidylcholine, hydrolyzed with phospholipase A _2, 1-acyl - after obtaining a phospholipid, obtained by transesterification using pomegranate seed oil or punicic acid with phospholipase A ₂ I can do it.

The glyceride of the present invention obtained as described above has a function of suppressing fat accumulation, a function of accelerating the reduction of accumulated fat, and a function of improving abnormal carbohydrate metabolism. As a result, obesity-based diabetes, hypertension, lipid metabolism The effect of preventing and improving lifestyle-related diseases such as abnormalities can be obtained. Since these lifestyle-related diseases are basic diseases of arteriosclerosis, heart disease and central disease, the glyceride of the present invention is expected to prevent and improve arteriosclerosis, heart disease and central disease.

The glyceride of the present invention can be used as an oil / fat composition by fractionation or mixing with other edible oil / fat as appropriate, or mixing with other edible oil / fat followed by transesterification. Other edible oils and fats are not particularly limited as long as they are oils ordinarily used for food, for example, butter, butter oil, milk fat such as cream, soybean oil, rapeseed oil, coconut oil, cottonseed oil, palm oil, Vegetable oils and fats such as corn oil, safflower oil and rice oil, and animal oils and fats such as lard, beef tallow, whale oil and sardine oil can be used alone or in combination.

The oil / fat composition of the present invention has no problem even if it contains additives usually added to the edible oil / fat composition in addition to the glyceride and the edible oil / fat. For example, water, milk, an emulsifier, an emulsion stabilizer, an antioxidant, a thickener, a flavoring agent, a flavoring agent and the like can be mentioned.

Uses of the glyceride-containing oil / fat composition of the present invention include foods, medicines and feeds. The food using the glyceride-containing oil / fat composition of the present invention is not particularly limited as long as it contains an oil / fat as a component. When it is expected that the glyceride of the present invention is contained in the food in an amount of 1% or more. If the amount is less than 1%, the functions of suppressing fat accumulation, accelerating the reduction of accumulated fat, and improving the abnormal function of carbohydrate metabolism may be insufficient. The form of the food is not particularly limited, and specific examples include canned salad oil, frying oil, margarine, shortening, cream, bread, cakes, cookies, frozen desserts, noodles, oil sardines and tuna-flavored flakes. , Pickle liquid, sauce, dressing, mayonnaise,
Examples include noodles, roux, cheese food, cheese-like food, and the like, and also include those obtained by secondary processing. For example, fried food, hamburger, curry, stew, croquette, fried rice, potato salad and the like can be mentioned. When producing these foods, the oil and fat composition of the present invention may be used in each food production process in the same manner as ordinary edible oils and fats.

The food using the oil or fat composition of the present invention contains glycerides having a function of suppressing fat accumulation, a function of accelerating the reduction of accumulated fat, and a function of improving abnormal carbohydrate metabolism. , As a food for the prevention and improvement of lifestyle-related diseases, as well as a health food for maintaining and promoting health,
It can be used as a dietary supplement.

Pharmaceuticals using the glyceride-containing oil / fat composition of the present invention have a function of suppressing fat accumulation, a function of accelerating the reduction of accumulated fat, and a function of abnormal carbohydrate metabolism, and prevent and improve lifestyle-related diseases based on obesity. Has an effect. When the oil / fat composition of the present invention is used as an active ingredient of a drug, it is used as it is or in a pharmaceutically acceptable non-toxic and inert carrier. In that case, the content of the glyceride of the present invention is preferably 1% or more for exhibiting the above function. As a carrier, solid, semi-solid, and liquid pharmaceutical preparations can be used in a usual pharmaceutical preparation, and are not particularly limited. However, excipients such as glucose and lactose, starch, carboxymethylcellulose calcium ( Disintegrators such as CMC-Ca), hydroxypropylcellulose (HPC), polyvinylpyrrolidone (PV
A binder such as P), a lubricant such as talc and magnesium stearate, a pH adjuster such as sodium bicarbonate, a stabilizer, a diluent, a dye, and one or more auxiliary agents for other formulations are used. The medicament of the present invention includes tablets, capsules, granules, powders, powders, dragees, suspensions, solutions, syrups, drops,
It can be used as an oral preparation such as sublingual tablets and emulsifiers, or as a parenteral preparation such as injections and suppositories. If long-term administration is required, oral administration is preferred. The medicament of the present invention can be used for humans and animals.

A feed using the glyceride-containing oil / fat composition of the present invention is a feed having a function of suppressing fat accumulation, a function of accelerating the reduction of accumulated fat, and a function of improving abnormal carbohydrate metabolism. It can be used as feed for food. The glyceride content of the present invention is preferably 1% or more. In the feed of the present invention, in addition to the oil composition,
It can contain compositions that are usually added to feed.

[0040]

The present invention will be described below in detail with reference to examples. It goes without saying that the present invention is not limited by the present embodiment. In the following examples, “parts” means parts by weight unless otherwise specified.

Example 1 Production of Glyceride Containing Conjugated Triene Highly Unsaturated Fatty Acid and Short-Chain Fatty Acid (1) 100 parts of dehydrated pomegranate seed oil and 50 parts of valeric acid methyl ester were placed in a reaction vessel, and sodium methoxide was added. One part was added and the mixture was stirred under vacuum at 80 ° C. for 30 minutes. After the reaction,
After adding 50 parts of water to deactivate the catalyst, extraction with an organic solvent was performed.
Silica gel column chromatography was performed to obtain a triglyceride fraction. After the triglyceride fraction was decolorized with activated carbon, the solvent was distilled off, and the fatty acid composition was analyzed by a conventional method. Methyl esterification is the standard test method for fats and oils (edited by the Japan Society of Fats and Fats)
It was carried out according to. The separation conditions for gas chromatography (GC) are as follows. 1) Column: capillary column, TC-70 (inner diameter 0.25 mm x length 30 m) (GL Science) 2) Flow rate: 0.8 ml / min, column head pressure: 100 k
Pa 3) Carrier gas: nitrogen gas 4) Column temperature: heating mode, 170-220 ° C (4 ° C)
/ Min) 5) Detection: = FID As a result, the obtained triglyceride contained 50 punicic acid residues.
%, Valeric acid residues 30%, and the rest were composed of fatty acid residues derived from pomegranate seed oil.

(Example 2) Production of glyceride containing conjugated triene highly unsaturated fatty acid and short chain fatty acid (2) Triglyceride was obtained in the same manner as in Example 1 except that bitter melon seed oil was used instead of pomegranate seed oil. Was. The resulting triglyceride contains 32% of α-eleostearic acid residue.
And 28% of valeric acid residues, and the remainder consisted of fatty acid residues derived from bitter melon seed oil.

Example 3 Production of Glyceride Containing Conjugated Triene Highly Unsaturated Fatty Acid and Short-Chain Fatty Acid (3) Triglyceride was obtained in the same manner as in Example 1 except that calendula seeds were used instead of pomegranate seed oil. . The obtained triglyceride was composed of 23% of calendic acid residues and 23% of valeric acid residues, and the remainder was composed of fatty acid residues derived from calendula seed oil.

Example 4 Production of Glyceride Containing Conjugated Triene Polyunsaturated Fatty Acid and Short-Chain Fatty Acid (4) Triglyceride was prepared in the same manner as in Example 1 except that american cattle seed oil was used instead of pomegranate seed oil. Got. The resulting triglyceride is a catalpic acid residue 22
%, Valeric acid residues 22%, and the remainder consisted of fatty acid residues derived from American cattle seed oil.

Example 5 Production of Glyceride Containing Conjugated Triene Highly Unsaturated Fatty Acid and Medium Chain Fatty Acid (1) 100 parts of pomegranate seed oil and 200 parts of n-hexane, 100 parts of palm kernel oil (lauric acid content 48%) Part was stirred and dissolved, and a 1,3-specific lipase (trade name: Lipozyme IM)
10 parts (manufactured by Novo Nordisk) were added, and the mixture was gently stirred at 50 ° C. for 24 hours under a nitrogen stream. After the reaction,
The lipozyme IM was separated by filtration, and the obtained reaction product was subjected to dehydration treatment, fractionated crystallization at 10 ° C. and then at −3 ° C., and the fatty acid composition of the obtained triglyceride was analyzed in the same manner as in Example 1. As a result, triglyceride obtained at 10 ° C. was 42% lauric acid, 20% punicic acid, 8.0% myristic acid,
The glyceride obtained at 8.2% oleic acid and -3 ° C. is 45% punicic acid, 21% lauric acid, 4.2 myristic acid.
%, Oleic acid 3.9%. Since the melting point of palm kernel oil is 27.3 ° C, the melting point of pomegranate seed oil is -5.5 ° C, and the enzyme used is 1,3-specific lipase, the obtained glyceride is 10 ° C. The main component of the triglyceride obtained in (1) is LPL (L indicates a lauric acid residue and P indicates a punicic acid residue), and the main component of the triglyceride obtained at −3 ° C. is PLP (L is a lauric acid residue). , P represents a punicic acid residue).

Example 6 Production of Glyceride Containing Conjugated Triene Highly Unsaturated Fatty Acid and Medium-Chain Fatty Acid (2) In the same manner as in Example 5, by using bitter melon seed oil instead of pomegranate seed oil, α -A triglyceride containing 31% eleostearic acid residues and 19% lauric acid residues was obtained.

Example 7 Production of Diglyceride Containing Conjugated Triene Polyunsaturated Fatty Acid 100 parts of pomegranate seed oil was dissolved in 100 parts of n-hexane, 1 part of 5 mM phosphate buffer, and 1 part of Lipozyme IM 0.1.
After adding 1 part and reacting at 40 ° C. for 6 hours, added lipozyme I
After removing M and dehydrating, eluting with hexane / diethyl ether using a silica gel column to separate a punicic acid fraction and a monoglyceride fraction, distilling off the solvent of each fraction, and adding 40 parts of punicic acid And 35 parts of monoglyceride were obtained. After dissolving 20 parts of the obtained monoglyceride and 20 parts of punicic acid in n-hexane, 0.1 part of lipozyme IM was added, and the mixture was reacted at 40 ° C. for 12 hours under a nitrogen stream, and the lipozyme IM was filtered off. Was purified using a silica gel column, and a diglyceride fraction was collected. Punicic acid residue is 75
%Met.

Example 8 Production of Glyceride Containing Conjugated Triene Polyunsaturated Fatty Acid and Conjugated Linoleic Acid 100 parts of active linol containing 70% of conjugated linoleic acid (manufactured by Linol Fats, Inc.) and 100 parts of pomegranate seed oil To n
-After dissolving in 200 parts of hexane, Lipozyme IM1
Then, the mixture was stirred gently at 40 ° C. for 24 hours to carry out a reaction. After completion of the reaction, lipozyme IM was removed by a conventional method, the reaction mixture was washed with an alkali to remove fatty acids, and 48 parts of triglyceride was recovered by silica gel chromatography. The fatty acid composition of the recovered triglyceride was analyzed. As a result, it was found that it was composed of punicic acid 48% and conjugated linoleic acid 22%.
As a result, PCP (here, PP
Represents punicic acid and C represents conjugated linoleic acid).

Example 9 Production of Glycerides Containing Conjugated Triene Highly Unsaturated Fatty Acid and Long Chain Saturated Fatty Acid 100 parts of behenic acid (manufactured by Wako Pure Chemical Industries, Ltd.) and 100 parts of pomegranate seed oil were treated with 200 parts of n-hexane. After dissolving in 1 part, 1 part of lipozyme IM was added, and the mixture was gently stirred at 60 ° C. for 12 hours to carry out a reaction. After completion of the reaction,
Lipozyme IM was removed, the reaction mixture was washed with alkali to remove fatty acids, and 40 parts of triglyceride were recovered by silica gel chromatography. The fatty acid composition of the recovered triglyceride was analyzed. As a result, it was 45% of punicic acid and 22% of behenic acid. As a result, PBP (where P represents punicic acid and B represents behenic acid) was obtained as the main component by the above reaction.

Example 10 Production of Glyceride Containing Conjugated Triene Polyunsaturated Fatty Acid and Docosahexaenoic Acid (1) 100 parts of docosahexaenoic acid ethyl ester (95% DHA ethyl ester, manufactured by Harima Chemicals, Inc.), pomegranate seeds After dissolving 100 parts of oil in 200 parts of n-hexane, 1 part of lipozyme IM was added, and 40 parts of the oil was added under a nitrogen stream.
The reaction was carried out by gentle stirring at 24 ° C. for 24 hours. After completion of the reaction, lipozyme IM was removed by a conventional method, the reaction mixture was washed with an alkali to remove fatty acids, and silica gel chromatography was used to recover 51 parts of triglyceride. The fatty acid composition of the recovered triglyceride was analyzed. As a result, punicic acid 53%, docosahexaenoic acid 2
3%. As a result, PDP (here, D represents docosahexaenoic acid and P represents punicic acid) was obtained by the above reaction.

Example 11 Production of Glyceride Containing Conjugated Triene Polyunsaturated Fatty Acid and Docosahexaenoic Acid (2) Instead of Lipozyme IM, Pseudomonas sp.
1 g of powdery lipase (PSL) derived from 56 strains (manufactured by Kurita Kogyo KK: hydrolysis activity = 56.5 U / mg) was added, and the reaction was carried out in the same manner as in Example 10. Glyceride was obtained from the obtained reaction product, and the fatty acid composition was analyzed. As a result, punicic acid was 49% and docosahexaenoic acid was 19%.

Example 12 Production of Glyceride Containing Conjugated Triene Highly Unsaturated Fatty Acid and Docosahexaenoic Acid (3) Instead of ethyl docosahexaenoate, punicic acid prepared in Example 7 was used instead of pomegranate seed oil Docosahexaenoic acid-containing triglyceride (DHA-70)
M) (Harima Chemical Industry Co., Ltd.), and the reaction was carried out in the same manner as in Example 10. As a result, a triglyceride containing 22% of punicic acid and 18% of docosahexaenoic acid was obtained.

Example 13 Production of Phospholipid Containing Conjugated Triene Polyunsaturated Fatty Acid 1 g of phosphatidylcholine (purified by egg yolk) having a purity of 95% (manufactured by Wako Pure Chemical Industries) was dissolved in 5 ml of n-hexane, and phospholipase A ₂ derived from actinomycete was used. Sigma) 10 mg,
After reacting at 40 ° C. for 12 hours, the reaction product was dehydrated, applied to a silica gel column, and developed with chloroform: methanol: acetone (83: 16: 1) to obtain a lysophosphatidylcholine fraction eluted later than phosphatidylcholine. Then, the solvent was distilled off to obtain 200 mg of lysophosphatidylcholine. After dissolving 200 mg of the obtained lysophosphatidylcholine and 200 mg of punicic acid prepared in Example 7 in n-hexane, ₂ m of the above phospholipase A2 was dissolved.
After reacting under vacuum at 40 ° C. for 12 hours, the product was formed on a silica gel column, the phosphatidylcholine fraction was collected, the solvent was distilled off, and phosphatidylcholine 100
mg was obtained. The obtained phosphatidylcholine was methylesterified by a boron trifluoride methanol method, and the fatty acid composition was measured by a conventional method. As a result, punicic acid was 31%,
Palmitic acid, stearic acid and oleic acid accounted for about 10% each.

(Example 14) Inhibition of fat accumulation Six-week-old female ICR CD-1 mice (Charles River Japan Co., Ltd.) were divided into eight mice / group, one group for mice and rats from which fat was removed. Standard formula feed (AIN for growing season)
-93G, Oriental Yeast Co., Ltd.) with 10% soybean oil (control group 1), and another group with 8% pomegranate seed oil and 2% soybean oil (test group). 1) Still another group was bred for 4 weeks with a feed containing 8% of glyceride PLP containing punicic acid and lauric acid produced in Example 5 and 2% of soybean oil (test group 2) for 4 weeks. Four weeks later, the animals were dissected under ether anesthesia, and the peri-renal adipose tissue and the peri-ovarian adipose tissue were collected and weighed. The obtained sum of both adipose tissue weights was divided by the body weight to obtain a fat tissue weight ratio to body weight. The results are shown in Table 1 as relative ratios when the weight ratio of adipose tissue to body weight of control group 1 was set to 100. As a result, Test Group 1, which took pomegranate seed oil and the glyceride containing punicic acid and lauric acid of the present invention,
Visceral fat accumulation was suppressed in 2 compared with the control group 1, and the effect was more remarkable in the glyceride group containing punicic acid and lauric acid (test group 2).

[0055]

[Table 1]

Using the glycerides obtained in Examples 1 to 4 and 6 to 12 and the phospholipid obtained in Example 13, the effect of suppressing fat accumulation was examined in the same manner as described above. The effect was recognized.

(Example 15) Visceral fat reducing effect A 10-week-old female C57BL / 6J mouse (manufactured by Nippon Charles River Co., Ltd.) was purified from a high-fat, high-sugar fractionated purified feed having the composition shown in Table 2 (Oriental Yeast Kogyo Co., Ltd.) After 4 weeks of breeding, the animals were divided into 8 animals / group, one group was dissected, and the peri-visceral adipose tissue and the peri-ovarian adipose tissue were collected and weighed (control group 2). The other group is
Fat-free standard formula feed for growing season (AIN-93)
G: a feed (test group 3) containing 2% of soybean oil and 8% of diglyceride containing punicic acid prepared in Example 7 as essential fatty acid sources (manufactured by Oriental Yeast Industry Co., Ltd.), or 2% of soybean oil and pomegranate They were bred for an additional 4 weeks on a feed supplemented with 8% of seed oil (test group 4). Four weeks later, the animals were dissected under ether anesthesia, and the peri-renal adipose tissue and the peri-ovarian adipose tissue were collected and weighed. The obtained sum of both adipose tissue weights was divided by the body weight to obtain a fat tissue weight ratio to body weight. The results are shown in Table 3 as a relative ratio when the weight ratio of adipose tissue to body weight of control group 2 was set to 100. As a result, the feed group (test group 3) to which 8% of diglyceride containing punicic acid was added showed a more remarkable visceral fat reduction promoting effect than the pomegranate seed oil group (test group 4).

[0058]

[Table 2]

[0059]

[Table 3]

Further, the visceral fat reducing effect was examined in the same manner as described above using each of the glycerides obtained in Examples 1 to 6 and 8 to 12 and the phospholipid obtained in Example 13, and The effect was recognized.

(Example 16) Effect of glyceride containing punicic acid and lauric acid on improvement of abnormal carbohydrate metabolism Five-week-old female diabetic KK-Ay mouse (average weight 27.5 g) (from CLEA Japan, Inc.) Was preliminarily reared for 1 week, divided into 5 animals / group, and one group (control group) was AIN-93G feed from which fat was removed (manufactured by Oriental Yeast Co., Ltd.) (casein 20.0%, corn starch 4).
9.948%, sucrose 10.0%, cellulose powder 5.0%, AIN-93 mineral mixture 3.5
%, AIN-93 vitamin mixture 1.0%, choline bitartrate 0.25%, tert-butyl hydroquinone 0.002%,
Modified AIN-93G feed with 10% soybean oil added to 0.30% L-cystine (energy ratio; 22% fat, 58.5% carbohydrates, 19.5% protein, 1 total energy)
7154 kJ / kg), and one group (troglidazone group) was a modified AIN-93G diet supplemented with 0.2% of an insulin sensitizer troglidazone, and another group (punicic acid and lauric acid-containing glyceride). Group)
The modified AIN-93G feed was bred for an additional 4 weeks under free-feeding conditions with a feed obtained by replacing 8% of the soybean oil with the glyceride PLP containing punicic acid and lauric acid prepared in Example 5.
The feed was replaced every two or three days with fresh ones and the food intake was recorded. In addition, blood was collected from the tail vein every week, and the blood glucose level during satiation was measured using a simple blood glucose meter (Novo Assist Plus: manufactured by Novo Nordex Farm Co., Ltd.).
The food consumption of each group during the test period averaged 5.
At 3 g / day, no significant difference was observed between the groups. No significant difference was observed between the groups in body weight. As a result, in the control group, the blood glucose level was 400 weeks after the start of the test.
The patient developed diabetes in excess of mg / dl and the condition persisted for up to 4 weeks. On the other hand, in the troglidazone group and the glyceride group containing punicic acid and lauric acid, the blood glucose level was maintained at 200 mg / dl or less throughout the test period, and remained almost at a normal value.

Using the glycerides obtained in Examples 1 to 4 and 6 to 12 and the phospholipid obtained in Example 13, the effect of improving the abnormal carbohydrate metabolism was examined in the same manner as described above. Similar effects were observed.

Example 17 Production of Margarine 80 parts of a fat and oil composition composed of 60% hydrogenated soybean oil (mp40 ° C.), 20% palm oil and 20% corn oil, 0.3 part lecithin, 0 glycerin fatty acid ester .3 parts, water 16
Parts, 2 parts of sodium chloride, 5 parts of glyceride containing punicic acid and docosahexaenoic acid prepared in Example 11, a small amount of vitamin E as an antioxidant, and TK homomixer (special equipment department) at 60 ° C. while blowing nitrogen gas. The mixture was emulsified at 50 V for 15 minutes using an industry (manufactured by Kogyo Co., Ltd.), and then rapidly cooled and kneaded at 15 ° C. to obtain a sheet-like margarine having no problem in flavor.

Example 18 Preparation of Oral Capsule After mixing 40 mg of triglyceride containing punicic acid and conjugated linoleic acid, 200 mg of lactose, 70 mg of starch, 50 mg of polyvinylpyrrolidone and 35 mg of crystalline cellulose obtained in Example 8, , # 3 gelatin capsules, and the surface was coated with gelatin to prepare oral capsules.

Example 19 Preparation of Oral Tablet 5 g of the triglyceride containing punicic acid and conjugated linoleic acid obtained in Example 8, 70 g of lactose and 30 g of corn starch were uniformly mixed, and 10% of 25 ml of a propylcellulose solution was added, followed by stirring and granulation.
After drying, this is sized and 2 g of magnesium stearate,
2 g of talc was added and mixed, and tablets could be produced with a rotary tableting machine.

Example 20 Preparation of Pet Food 80 parts chicken surimi, 10 parts minced lean beef, 1 soy protein
0 parts, 3 parts of triglyceride LPL containing punicic acid and lauric acid produced in Example 5, 1 part of chemical seasoning, 1 part of tocopherol, a little calcium, a few vitamins, a little starch, a little sorbitol with a food cutter It was kneaded, filled into sheep intestine, cooked at 90-95 ° C., and then air-dried at 50 ° C. to obtain a dry sausage type pet food.

Example 21 Stability Test of Glyceride The triglyceride containing punicic acid and valeric acid obtained in Example 1 and the LPL obtained in Example 5 and pomegranate seed oil containing punicic acid triglyceride as a main component Stability
Using an automatic oil / fat stability tester Rancimat 679 (manufactured by Metrohm Shibata Co., Ltd.) at a temperature of 80 ° C. and a ventilation rate of 1
The induction time was compared under the condition of 0 L / hour.
As a result, with respect to 5.5 hours of pomegranate seed oil, triglyceride containing punicic acid and valeric acid was 7 hours, LPL was 7.5 hours, and punicic acid and valeric acid were contained as compared with pomegranate seed oil. The improved stability was observed for triglycerides and LPL. In addition, as a result of a sensory test of the flavor after leaving these fats and oils at room temperature for one month, an improvement in the flavor was observed in triglyceride and LPL containing punicic acid and valeric acid as compared with pomegranate seeds.

[0068]

The glyceride of the present invention has a superior function of suppressing fat accumulation, a function of accelerating reduction of accumulated fat, and a function of ameliorating abnormal carbohydrate metabolism, as compared with glycerides comprising conjugated polyunsaturated fatty acids having a conjugated triene structure derived from nature. It is useful as a health food, medicine, and feed for preventing and improving lifestyle-related diseases such as diabetes, hypertension, and abnormal lipid metabolism based on obesity. .

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme court ゛ (Reference) A61K 31/232 A61K 31/232 A61P 3/04 A61P 3/04 3/06 3/06 3/10 3 / 10 9/00 9/00 9/10 101 9/10 101 9/12 9/12 25/28 25/28 C11B 11/00 C11B 11/00 C11C 3/08 C11C 3/08 3/10 3/10 F Term (Reference) 2B150 AA06 AB03 BC01 CJ08 DA37 DA55 DA58 DC08 DH14 4B018 LB01 LE01 MD15 ME01 ME03 MF10 4B026 DC05 DG01 DH01 DP03 4C206 AA01 AA02 AA04 DB09 DB47 DB48 MA01 MA04 NA14 ZA02 ZA36 ZA42 ZA33 BC33B33B33C CA35 CA37 EA17

Claims (11)

[Claims] 1. A conjugated polyunsaturated fatty acid having one or two conjugated polyunsaturated fatty acids having a conjugated triene structure, or a conjugated polyunsaturated fatty acid having a conjugated triene structure,
5, short-chain fatty acids having 5 to 12 carbon atoms, medium-chain fatty acids having 6 to 12 carbon atoms, long-chain saturated fatty acids having 20 to 22 carbon atoms, fatty acids having a conjugated diene structure having 18 carbon atoms, and having a degree of unsaturation of 3 to 18 having 22 to 22 carbon atoms.
6 highly unsaturated fatty acids and -PO ₃ groups represented by H-B of [B choline, serine, or shows the ethanolamine or inositol radical glycerides containing at least one selected from the group consisting of . 2. TAT, TTA, ATA, THT, TA
P, TTP or THP [T is a conjugated polyunsaturated fatty acid residue having a conjugated triene structure, A is a short chain fatty acid residue having 2 to 5 carbon atoms, a medium chain fatty acid residue having 6 to 12 carbon atoms, and 20 carbon atoms. Any one selected from a long-chain saturated fatty acid residue having a conjugated diene structure having 18 carbon atoms, a highly unsaturated fatty acid residue having 18 to 22 carbon atoms and a degree of unsaturation of 3 to 6; hydrogen atom, P is -PO ₃ H-B (B is choline,
Glyceride according to claim 1, which represents any of serine, ethanolamine and inositol residues)]. 3. The conjugated polyunsaturated fatty acid having a conjugated triene structure comprises punicic acid, α-eleostearic acid, β-
The glyceride according to claim 1 or 2, wherein the glyceride is at least one selected from the group consisting of eleostearic acid, jalcalic acid, calendic acid and catalpic acid. 4. The polyunsaturated fatty acid having 18 to 22 carbon atoms and a degree of unsaturation of 3 to 6 is selected from the group consisting of eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, γ-linolenic acid and α-linolenic acid. At least one
A glyceride according to claim 1 or 2 which is a species. 5. The glyceride according to claim 1, which has at least one of a function of suppressing fat accumulation, a function of accelerating reduction of accumulated fat, and a function of improving abnormal carbohydrate metabolism. 6. A conjugated polyunsaturated fatty acid having a conjugated triene structure or an ester derivative thereof, a short-chain fatty acid having 2 to 5 carbon atoms, a medium-chain fatty acid having 6 to 12 carbon atoms, and 20 carbon atoms.
-22 long-chain saturated fatty acids, fatty acids having a conjugated diene structure having 18 carbon atoms, polyunsaturated fatty acids having 18-22 carbon atoms and a degree of unsaturation of 3-6, ester derivatives thereof, and phospholipids. 6. A transesterification reaction with at least one of the above compounds.
The method for producing glyceride according to any one of the above. 7. An ester derivative of a conjugated polyunsaturated fatty acid, comprising: pomegranate seed oil, bitter melon seed oil, calendula seed oil, rosewood seed oil, catalpa seed oil, American cattle seed oil, balsam apple seed oil, snake guard seed oil, The method for producing glyceride according to claim 6, which is a glyceride obtained from at least one seed oil selected from the group consisting of oilseed seed oil and cherry oil. 8. A composition comprising at least one of the glycerides according to claim 1 and at least one of a function of suppressing fat accumulation, a function of accelerating reduction of accumulated fat, and a function of improving abnormal carbohydrate metabolism. An oil / fat composition having a function. 9. A food comprising the oil / fat composition according to claim 8. 10. A pharmaceutical comprising the oil / fat composition according to claim 8. 11. A feed comprising the oil / fat composition according to claim 8.