JP2002265985A - Lipid composition for inhibiting secretion of apolipoprotein-b - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide the use of a now found lipid composition which more strongly inhibits the secretion of apolipoprotein B from hepatocytes than α- linolenic acid as its foods, pharmaceuticals or animal feeding stuff. SOLUTION: The lipid composition for inhibiting the secretion of an apolipoprotein comprises at least one kind to be selected from the group consisting of a conjugated triene acid type fatty acid, its salt, and its derivative, and more strongly inhibits the secretion of apolipoprotein B than α-linolenic acid.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to apolipoprotein B
The present invention relates to a lipid composition characterized by suppressing secretion of apo B (hereinafter abbreviated as apo B). More specifically, the present invention provides a lipid comprising at least one member selected from the group consisting of conjugated trienoic acid-based fatty acids, salts thereof, and derivatives thereof, which suppress apoB secretion more strongly than α-linolenic acid. The present invention relates to a composition and its use as a food, medicine, feed.

[0002]

2. Description of the Related Art Apolipoprotein is a special plasma protein constituting lipoprotein, and has a function of solubilizing lipids in blood and transporting them to various organs. Apolipoprotein plays a role in activating and suppressing enzymes involved in lipid metabolism, and is an important plasma component that regulates lipid metabolism.
Apo B, a main component of apolipoprotein, is synthesized in the liver and associates with apolipoprotein E, apolipoprotein C, triglyceride (hereinafter abbreviated as TG), phospholipid, cholesterol ester (hereinafter abbreviated as CE), and the like. It forms an ultra-low-density lipoprotein (hereinafter abbreviated as VLDL) and transports TG and CE synthesized in the liver to peripheral tissues. ApoB secretion is enhanced by VLD in the blood.
It causes an increase in L concentration, an increase in low-density lipoprotein (hereinafter abbreviated as LDL) concentration, an increase in blood TG, and an increase in blood CE concentration, causing so-called hypertriglyceridemia and hypercholesterolemia. It has become one of. Apo B is one of the ligands of the scavenger receptor on the macrophage cell membrane. It is said that macrophages recognize oxidized LDL via apo B and actively phagocytose, thereby being involved in the development of atherosclerosis in arteries. Increased blood TG, increased blood CE levels, and atherosclerosis further increase obesity, diabetes,
It is regarded as an important risk factor for inducing lifestyle-related diseases such as hypertension, coronary artery disease, and cerebral artery disease. Therefore, by suppressing apo B secretion, blood CE, T
It is expected to reduce G concentration and LDL concentration in blood to prevent and improve lifestyle-related diseases such as atherosclerosis, obesity, diabetes, hypertension, coronary artery disease and cerebral artery disease. Is done. In fact, regarding the clinical effects of lowering blood LDL levels, large-scale clinical trials conducted on patients with abnormal lipid metabolism and glucose metabolism could significantly reduce the incidence of coronary artery disease. (Taskinen MR.,
Smith V. , J. et al. Intern. Med. , 24
Ritsu, Kawamura, Modern Fisher, 20 (10), 1241, 2000). In addition, increased secretion of apo B is considered to be associated with the onset and exacerbation of chronic nephritis, nephrosis, cirrhosis, and obstructive jaundice.

[0003] Lipids are important nutrients as an energy source and as a constituent of living organisms. Saturated fatty acids or monoene fatty acids, which are the main constituents, have the property of enhancing apo B secretion. Atherosclerosis, obesity, hypercholesterolemia, hypertriglyceridemia, fatty liver, lifestyle-related diseases represented by diabetes or hypertension, coronary artery disease, cerebral artery disease, chronic nephritis, nephrosis, liver cirrhosis, obstructive jaundice It is suspected that it is involved in the onset and exacerbation of the disease, and it is recommended to refrain from taking it for health. However, lipids are not only important nutrients for mammals as described above, but also important components for imparting delicious taste such as taste, umami, and kokumi to foods. Nutrition,
It is not preferable in terms of physical properties and flavor. As a method for solving this problem, a method using a lipid which is a lipid and has a characteristic of suppressing apoB secretion can be considered.

[0004] Lipids having the property of inhibiting apo B secretion include α-linolenic acid and 10 trans, 12 ci.
s-Linoleic acid (T. Yanagita et.a.
l. , Food Res. Intern. , 31
(5), 403, 1999), but the degree of suppression of apoB secretion by these is not sufficient, and there is a need for a lipid having a strong effect of suppressing apoB secretion. Examples of substances other than lipids include taurine and γ-oryzanol (Yanagida et al., Abstracts of 98th Annual Meeting of the Japanese Society of Agricultural Chemistry, 3E
2a10), sulfur-containing aminosulfoxide derived from onion (Yanagida et al., Abstracts of the 2000 Annual Meeting of the Japanese Society for Agricultural Chemistry, 2G020α), NK-104 (3-Hydrox)
y-3-Methlyglutayl Coenzym
eA Reduced Inhibitor) (Y
anagita et. al. , Current Th
erapeutic Research, 60,
(8), 423, 1999), anhydrous 4'-trifluoromethylbiphenyl-2-carboxylic acid hydrochloride- [2- (2
-Acetylaminoethyl) -1,2,3,4-tetrahydroisoquinolin-6-yl] amide (JP-A-11-6
0557) 4′-trifluoromethyl-biphenyl-
2-carboxylic acid [2- (1H- [1,2,4-triazol-3-ylmethyl)]-(1,2,3,4-tetrahydro-isoquinolin-6-yl) -amide (Table 1)
2-510483), biphenyl-2-carboxylic acid-tetrahydro-isoquinolin-6-ylamide derivative (Japanese Patent Application Laid-Open No. 11-514964), etc., are more effective and safer, and can be used as foods, medicines or feeds. There is a demand for the development of a component that can be easily applied to water.

[0005]

An object of the present invention is to find a lipid composition which suppresses apo B secretion from hepatocytes more strongly than α-linolenic acid, and to provide its use as a food, medicine or feed. It is in.

[0006]

Means for Solving the Problems The present inventors have, conventionally,
I have been studying conjugated trienoic acid-based fats and oils. During a series of studies, apoB in hepatic cells of conjugated trienoic acid
When the secretion inducing amount was measured, it was found that α-linolenic acid, which is already known to suppress apo B secretion, was significantly suppressed. As a result of further studies, the present invention was completed. Was.

That is, a first aspect of the present invention is an apolipoprotein B secretion-inhibiting lipid composition comprising at least one member selected from the group consisting of conjugated trienoic acid-based fatty acids, salts thereof, and derivatives thereof. About. In a preferred embodiment, apolipoprotein B is stronger than α-linolenic acid.
The present invention relates to the above-described apolipoprotein B secretion-inhibiting lipid composition, which comprises at least one selected from the group consisting of conjugated trienoic acid-based fatty acids, salts thereof, and derivatives thereof, which inhibit secretion.

In a further preferred embodiment, the conjugated trienoic acid is selected from the group consisting of punicic acid, calendic acid, jarcaric acid, α-eleostearic acid, β-eleostearic acid, and catalpic acid. 1
And at least one apolipoprotein B secretion-suppressing lipid composition described above. As another more preferred embodiment, the apolipoprotein B secretion-inhibiting lipid composition according to any one of the above, wherein the derivative of the conjugated trienoic acid-based fatty acid is an ethyl ester derivative or a glycerol ester derivative of the conjugated trienoic acid-based fatty acid About. In another more preferred embodiment, the glycerol ester derivative of a conjugated trienoic acid-based fatty acid comprises pomegranate seed oil, calendula seed oil, catalpa seed oil, bitter melon seed oil, balsam apple seed oil, abragiri seed oil, and cherry seed oil. The present invention relates to the aforementioned apolipoprotein B secretion-suppressing lipid composition, which is at least one member selected from the group.

A second aspect of the present invention relates to a food comprising the lipid composition for inhibiting apolipoprotein B secretion described in any of the above, and in a preferred embodiment, atherosclerosis, obesity, hypercholesterolemia The food according to the above, wherein the food is for prevention or improvement of at least one selected from the group consisting of hypertriglyceridemia, fatty liver, diabetes and hypertension.

A third aspect of the present invention relates to a pharmaceutical comprising the apolipoprotein B secretion-inhibiting lipid composition described in any of the above, and in a preferred embodiment, atherosclerosis, obesity, hypercholesterolemia And at least one preventive or ameliorating agent selected from the group consisting of hypertriglyceridemia, fatty liver, diabetes, and hypertension.

A fourth aspect of the present invention relates to a feed comprising the apolipoprotein B secretion-inhibiting lipid composition described in any of the above.

A fifth aspect of the present invention is to provide any of the above-mentioned apolipoprotein B secretion-inhibiting lipid compositions and apolipoprotein B secretion-inhibiting lipid compositions in a mammal in need of inhibiting apolipoprotein B secretion. Foods, medicines,
The present invention relates to a method for suppressing apolipoprotein B secretion, comprising ingesting at least one member selected from the group consisting of feed.

A sixth aspect of the present invention is selected from the group consisting of the apolipoprotein B secretion-inhibiting lipid composition according to any of the above, and a food, pharmaceutical, or feed comprising the apolipoprotein B secretion-inhibiting lipid composition. Preventing at least one selected from the group consisting of atherosclerosis, obesity, hypercholesterolemia, hypertriglyceridemia, fatty liver, diabetes or hypertension, characterized by ingesting at least one selected from the group consisting of: How to improve.

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION Apolipoprotein B of the present invention will be described below.
The secretion inhibiting lipid composition will be described in detail.

The apolipoprotein B secretion-inhibiting lipid composition of the present invention (hereinafter sometimes abbreviated simply as “lipid composition”)
Contains at least one or more selected from the group consisting of fatty acids having a conjugated trienoic acid structure in the molecule (hereinafter abbreviated as conjugated trienoic acid-based fatty acids), salts thereof, and derivatives thereof, and a liver per molecule. It has the characteristic of inhibiting apoB secretion from cells more strongly than α-linolenic acid. The conjugated triene-based fatty acid contained in the lipid composition of the present invention has a structure in which three or more unsaturated bonds can be conjugated in a molecule, and is a food, nutritional, physiological, or pharmaceutical agent. It is not limited by the number of carbon atoms, the number and position of other unsaturated bonds, the presence or absence of a substituent, etc., provided that the number of carbon atoms is 10 to 22 and the degree of unsaturation is 3 to 6.
Is preferably a conjugated trienoic acid-based fatty acid having 18 carbon atoms and derived from vegetable seed oil in terms of availability and the like. Specific examples of such conjugated trienoic acid fatty acids include punicic acid (1)
8: 3, 9c, 11t, 13c), calendic acid (ca
lendic acid) (18: 3, 8t, 10t, 12c),
Jarcaric acid (18: 3,8
c, 10t, 12c), α-eleostearic acid (α-ele
ostealic acid) (18: 3, 9c, 11t, 13t),
β-eleostealic acid (1
8: 3, 9 t, 11 t, 13 t), catalpic acid (cata
lpic acid) (18: 3, 9t, 11t, 13c), kamlolenic acid (18OH, 9c,
11t, 13t), parinaric acid
(18: 4, 9c, 11t, 13t, 15c) and other conjugated octadecatrienoic acid, eicosatetraenoic acid and the like. Among them, punicic acid, calendic acid,
Jalcalic acid, α-eleostearic acid, β-eleostearic acid, and catalpic acid are preferred, and punicic acid and α-eleostearic acid are most preferred in terms of stability and economy.

The salt of the conjugated triene fatty acid in the present invention is not particularly limited as long as it is food-, nutrient- or pharmacologically-acceptable, and metal salts such as sodium salt and calcium salt, Examples thereof include ammonium salts, salts with organic bases such as methylamine, ethylamine, diethylamine, triethylamine, pyrrolidine, piperidine, morpholine, hexamethyleneimine, aniline and pyridine, and salts with amino acids such as arginine, glutamic acid and ornithine. Further, as the derivative of the conjugated trienoic acid-based fatty acid in the present invention, any food-, nutritional-, or pharmacologically-acceptable one can be selected, but an ester derivative that is easily produced is preferable. Further, a methyl ester, an ethyl ester, a butyl ester, a propyl ester, and a glycerol ester are preferred, and an ethyl ester derivative and a glycerol ester derivative are most preferred. The glycerol ester derivative may be in any form of monoglyceride, diglyceride, and triglyceride, but diglyceride and triglyceride are preferred in view of physical properties as a food or pharmaceutical composition, and triglyceride is most preferred.
In these diglycerides and triglycerides, the position where the conjugated trienoic acid-based fatty acid is esterified may be selected according to the purpose, and is not particularly limited. Specific ester derivatives of conjugated trienoic acid fatty acids include ethyl punicate, ethyl calendate, ethyl jarcaricate, ethyl α-eleostearate, ethyl β-eleostearate, ethyl catalpinate, ethyl parinarate Butyl punicate, butyl α-eleostearate, propyl punicate, propyl α-eleostearate, 1-puniseal-sn-glycerol (1-punicyl)
-sn-glycerol), 2-punicyl-sn-glycerol, 1,2-dipniseal-
sn-glycerol (1,2-dipunicyl-sn-glycero
l), 1,3-dipunisyl-sn-glycerol (1,3-dipunicyl-sn-glycerol), 1,2,3-tripnisyl-sn-glycerol (1,2,3-tripunil)
cyl-sn-glycerol), 1-α-eleostaryl-s
n-glycerol (1-α-ereostearyl-sn-glycero
l), 2-α-ereostearyl-sn-glycerol, 1,2-diα
-Eleostaryl-sn-glycerol (1,2-d
iα-ereostearyl-sn-glycerol), 1,3-diα-eleostearyl-sn-glycerol (1,3-diα-er
eostearyl-sn-glycerol) 1,2,3-triα-eleostaryl-sn-glycerol (1,2,3-triα
-ereostearyl-sn-glycerol). Conjugated triene-based fatty acid contained in the lipid composition of the present invention, a salt thereof,
Derivatives can be prepared by chemical reaction or enzymatic reaction, etc., extracted from natural animals and plants, or obtained from natural animals and plants by chemical reaction or enzymatic reaction. Although various forms such as those changed to can be used, considering the use as food, medicine, or feed, those prepared from natural plants, animals, and marine products are preferable, and furthermore, securing of quantitative Roots, stems, leaves, flowers, seeds and the like of the plant are more preferred from the aspect.
Examples of these plants include the pomegranate (Punicaceae), the terrestrial pomegranate (Compositae (Asteraceae)),
Uphordiaceae), roots, stems, leaves, flowers or seeds of plants belonging to the family Cucuritaceae, the family Bignoniaceae, or the family Rosaeceae. Roots, stems, leaves, flowers, or seeds of plants belonging to pomegranates, roses, and cormorants, which are easily available quantitatively, are more preferable.
Some of these plants can be crushed, extracted and processed for use.

The method of crushing, extracting and processing the plant is as follows:
Apo B of conjugated trienoic acid fatty acids, salts and derivatives thereof
Any method can be selected from general methods as long as it does not adversely affect the secretion-inhibiting effect and the physical properties and flavor of food, medicine, or feed. For example, as a method of pulverizing, there are a method using a mortar, a blender, a mill, a homogenizer, a roller, a pressing machine, and the like, and a method of applying an impact in a frozen state. Examples of the extraction method include a method of extracting ground seeds and the like using an organic solvent such as n-hexane and acetone, and a method of performing solid-liquid separation using a pressing machine or the like.

In the present invention, among the extracts obtained from plants, seed oil extracted from seeds is preferred. Specific examples of preferred plant seed oils include pomegranate seed oil, calendula seed oil, rosewood seed oil, catalpa seed oil, American cattle seed oil, bitter gourd seed oil,
Balsam Apple Seed Oil, Snake Guard Seed Oil, Chikarasuuri Seed Oil, Pumpkin Seed Oil, Aburagiri Seed Oil,
And cherries seed oil and the like. Of these, from the content of conjugated trienoic acid fatty acids, salts and derivatives thereof in seeds, pomegranate seed oil, calendula seed oil, catalpa seed oil, bitter gourd seed oil, balsam apple seed Oil, oilseed seed oil and cherry seed oil are preferred, and pomegranate seed oil and bitter gourd seed oil are most preferred because of their availability.

The conjugated triene-based fatty acids, salts and derivatives thereof contained in the lipid composition of the present invention have a purity of the active ingredient and a process for producing the same if they have an effect of suppressing the secretion of apoB from hepatocytes. There is no particular restriction by For example, the active ingredient may be purified from natural raw materials to high purity by adsorption chromatography, partition chromatography, ion exchange chromatography, molecular sieve chromatography, fractional extraction with an organic solvent, fractional crystallization, molecular distillation, etc. good. Further, fatty acids or ester derivatives thereof may be obtained from plant seed oil. When a fatty acid is obtained, the above animal and vegetable fats and oils can be purified by a common method such as fractional extraction, column chromatography or the like after hydrolysis with alkali or lipase. The ester derivative of the conjugated triene-based fatty acid in the present invention is not particularly limited by its manufacturing method. For example, a direct esterification method of synthesizing the fatty acid with an alcohol such as ethyl alcohol or glycerol by a dehydration reaction, an ester and an alcohol Or a transesterification method of synthesizing a new ester by the reaction of an ester and the fatty acid or an ester and the ester, a method of synthesizing the fatty acid with a chloride and an alcohol, a method of reacting the epoxy compound with the fatty acid,
It can be prepared by a general method such as a method of reacting an olefin with the fatty acid.

The lipid composition of the present invention may contain, as a composition other than conjugated trienoic acid-based fatty acids, salts and derivatives thereof, an antioxidant, a deoxidizer, and a substance exhibiting an antioxidant action in a living body. . Antioxidants include, for example, lecithin, phospholipids such as phosphatidylethanolamine, tocopherols, ascorbic acid and ascorbic acid derivatives such as ascorbyl palmitate, vitamin A precursors such as carotene, polyphenols such as catechin, rutin Flavones, isoflavones, rosemary extract, etc.
Examples include plant extracts such as Pycnogenol, plant pigments such as lycopene, citrate esters such as isopropyl citrate, dibutylhydroxytoluene (BHT), propyl gallate, erythorbic acid, and sodium erysorbate. Lecithin, tocopherols, ascorbic acid palmitate and the like can be mentioned depending on the price and the strength of the antioxidant activity. Examples of the oxygen scavenger include inorganic oxygen scavengers such as iron powder and organic oxygen scavengers such as vitamin C and catechol. Examples of substances that exhibit an antioxidant effect in vivo include enzymes such as glutathione peroxidase, glutathione-S-transferase, catalase, peroxidase, superoxide desmutase, transferrin, ferritin, iron chelating agents such as lactoferrin, ceruloplasmin and the like. Copper ion chelating agent, β-carotene, vitamin E, vitamin C, ubiquinol, 2-hydroxyestradiol, uric acid, bilirubin, selenium, zinc and the like.

The lipid composition of the present invention may contain lipids other than conjugated trienoic acid-based fatty acids, salts and derivatives thereof.
The lipid to be contained is not particularly limited as long as the lipid composition of the present invention does not adversely affect the apo B secretion inhibitory effect, flavor and physical properties, and depending on the purpose of use, for example, safflower oil, kaya oil , Oils such as walnut oil, poppy oil, sunflower oil, semi-dry oils such as cottonseed oil, rapeseed oil, soybean oil, mustard oil, kapok oil, rice bran oil, sesame oil, corn oil, peanut oil, olive oil, camellia oil, tea Non-drying oils such as oil, coconut oil, palm oil, palm kernel oil, cocoa butter,
It can be selected from vegetable solid fats such as shea butter, borneo butter, animal fats and oils such as tallow, lard, sheep butter and butter.

The concentration of the conjugated trienoic acid-based fatty acid, a salt thereof, or a derivative thereof in the lipid composition of the present invention is not particularly limited as long as it is at least a concentration sufficient to suppress apoB secretion from hepatocytes. The conjugated trienoic acid-based fatty acid in the lipid composition may be 0.01% by weight (hereinafter abbreviated as%) or more, preferably 0.1% or more. If it is less than 0.01%, it is difficult to recognize a substantial apo B secretion inhibitory effect.

The effect of the lipid composition of the present invention on inhibiting apoB secretion is evaluated by a method of administering or ingesting the lipid composition of the present invention to mammals, or by allowing the lipid composition of the present invention to act on cultured hepatocytes. Although possible, the method using cultured hepatocytes is excellent in sensitivity, reproducibility and simplicity, and this method is usually used. In the case of the method using cultured hepatocytes, for example, human hepatocytes (HepG2)
After making the cells semiconfluent in a 24-well plate containing DMEM medium containing 0% fetal calf serum, the cells were transferred to DMEM.
After washing twice with a MEM medium, the medium was replaced with a DMEM medium containing 1% bovine serum albumin, and after pre-incubation for 24 hours, the medium was replaced with a DMEM medium containing 1% bovine serum albumin in which the lipid composition of the present invention was appropriately dissolved. After culturing for 24 hours or 24 hours, the culture solution is collected, and apoB contained in the culture solution may be quantified by sandwich ELISA (enzyme linked immunosorbent assay) (T.Yanagita et. al., Current Therapeut
ic Reserch., 60 (8), 423, 1999).

According to the above-mentioned method, it can be seen that the conjugated triene-based fatty acids, salts and derivatives thereof of the present invention suppress apoB secretion more strongly than α-linolenic acid.

The food containing the lipid composition for inhibiting apo B secretion of the present invention is compared with a food containing a normal lipid composition containing a normal saturated fatty acid or a monoenoic fatty acid as a main component. And has the property of inhibiting apo B secretion.
Foods containing the lipid composition of the present invention that inhibits apo B secretion include edible oils and fats such as salad oil, stir fry oil and frying oil, processed oil and fat foods such as margarine and shortening, mayonnaise, dressing, soy sauce, miso, Seasonings such as sauces, bakery products such as bread, cooked rice, noodles, processed soy foods such as tofu, processed fish and meat meat foods such as sausage and ham, cakes, cookies, buns, crackers, ice cream, puddings, yokan, candy, Examples include confectionery such as chocolate, dairy products such as butter, yogurt, and cheese, beverages such as coffee, black tea, green tea, oolong tea, sports drinks, konjac, and pickles.

The food containing the lipid composition of the present invention is
Contains various carriers and additives in addition to the lipid composition of the present invention
I can do it. The carriers and additives used are the lipid groups of the present invention.
As long as it does not adversely affect the apoB secretion inhibitory effect of the product
It may be. Specific examples of such a carrier include:
Seed carrier, extender, diluent, bulking agent
Agents, dispersants, excipients such as lactose, hydroxypropylcell
Loose (HPC), polyvinylpyrrolidone (PVP), etc.
Binders, solvents such as water, ethanol, vegetable oil, etc.
Agents, buffers such as baking soda, dissolution promoters, sodium CMC,
Gelling agents such as HPMC, agar and gelatin, sodium
Suspending agents such as mulginate and the like can be mentioned. Also,
Specific examples of the additive include glutamine soda, inosinic acid
Seasonings, banni to improve edibility and taste, such as
La, mint, rosemary, linalool, natural flavors, etc.
Fragrance, vitamin A, vitamin B₁, Vitamin B_Two, Bitami
B ₆, Vitamin C, vitamin E, pantothenic acid, nico
Vitamins such as formic acid, citric acid, malic acid, fumaric
Organic acids such as acid, malonic acid, succinic acid, tartaric acid, lactic acid, etc.
Colorants, moisture inhibitors, fibers, electrolytes, minerals, Sakae
Nutrients, antioxidants, preservatives, fragrances, humectants, tea extracts,
Coffee extract, cocoa extract, orange, grape,
Apple, peach, pineapple, pear, plum, cherry
Mombo, papaya, tomato, melon, strawberry, raspberry
-Natural plant extracts such as fruit extracts such as
It is.

The food of the present invention obtained as described above has a significantly lower ApoB secretion than a food using α-linolenic acid, which is known as an ApoB secretion inhibitory fat, and has a higher ApoB secretion. Prevention and improvement of atherosclerosis, obesity, hypercholesterolemia, hypertriglyceridemia, fatty liver, diabetes, hypertension, and chronic nephritis, nephrosis, cirrhosis, and obstructive jaundice based on hypersecretion It may be used as a target food. Moreover, you may use as a health food as a capsule or tablet which is not the form of normal food. The amount of the lipid composition of the present invention to be added to these foods is appropriately selected depending on the intended purpose, and the amount of the lipophilic fatty acid is preferably 0.01% or more, preferably 0.1% or more, as the conjugated trienoic fatty acid. What is necessary is just to add a lipid composition. If it is less than 0.01%, it is difficult to recognize a substantial apo B secretion inhibitory effect.

The lipid composition for inhibiting apoB secretion of the present invention can be used as a medicament for preventing / treating diseases based on enhanced apoB secretion. In that case, it may be used by blending it in a pharmaceutically acceptable non-toxic and inert carrier. In the medicament of the present invention, as the inert carrier that can be blended with the conjugated trienoic acid-based fatty acid, a salt thereof, or a derivative thereof, a solid, semi-solid, or liquid carrier used during normal production in a pharmaceutical preparation is used. You can do it. For example, excipients such as glucose, lactose, starch,
Carboxymethylcellulose calcium (CMC-C
a) and other disintegrants such as hydroxypropylcellulose (HP
C), binders such as polyvinylpyrrolidone (PVP), lubricants such as talc and magnesium stearate, pH regulators such as sodium bicarbonate, stabilizers, diluents, pigments, and other prescription auxiliaries. Can be used. The medicament of the present invention can be used as an oral preparation such as tablets, capsules, granules, powders, powders, dragees, suspensions, solutions, syrups, drops, sublingual tablets, emulsifiers, injections, suppositories, etc. And the like. If long-term administration is required, oral administration may be used.

The drug of the present invention thus obtained is
By significantly suppressing ApoB secretion, atherosclerosis, obesity, hypercholesterolemia, hypertriglyceridemia, fatty liver, diabetes, hypertension, and chronic nephritis based on apoB secretion enhancement , Nephrotic, cirrhosis, and obstructive jaundice.

When the oil or fat composition of the present invention is used as a drug, the fat or oil composition is used as a conjugated trienoic acid-based fatty acid in the drug.
It may be contained at least 01%, preferably at least 0.1%.
If it is less than 0.01%, it is difficult to recognize a substantial apo B secretion inhibitory effect. In addition, the daily intake or dosage is preferably 0.1 mg / kg / day or more, preferably 1 mg / kg / day or more, more preferably 10 mg / kg / day or more as a conjugated trienoic acid-based fatty acid. 0.1
If the amount is less than mg / kg / day, the effect of suppressing apoB secretion may not be observed.

The lipid composition for suppressing apo B secretion of the present invention may be used as a feed. In that case, the feed can be used as it is or by appropriately mixing seasonings, flavors and the like for improving edibility and palatability. In addition, in order to maintain certain physical properties, the feed can be formulated with an emulsifier and a stabilizer. Further, they can be used as raw materials for various processed feeds and pet foods produced industrially. In this case, the mammal can be used as a feed containing 0.01% or more, preferably 0.1% or more as a conjugated triene-based fatty acid with respect to the weight of the added feed or pet food. If it is less than 0.01%, the effect of suppressing apoB secretion may not be observed. The apo B of the present invention
The feed containing the lipid composition that suppresses secretion can be used as a feed for maintaining and improving the health of mammals as it is or by adding it to a normal feed. As the method of use, the lipid composition of the present invention may be added to a food material and cooked for use. You can also sprinkle it on your diet.

Feeds containing the lipid composition of the present invention include pets and other pets, and atherosclerosis in animals such as livestock, obesity, hypercholesterolemia, hypertriglyceridemia, fatty liver, diabetes, and hypertension. Furthermore, the prevention / amelioration of chronic nephritis, nephrosis, cirrhosis, and obstructive jaundice may be used as a feed for prevention / improvement.

Apolipoprotein B secretion can be suppressed by ingesting the lipid composition of the present invention into a mammal in which suppression of apolipoprotein B secretion is required. Furthermore, the method for inhibiting ApoB secretion of the present invention
By having the property of suppressing apoB secretion more effectively than α-linolenic acid, which is known to suppress secretion, atherosclerosis, obesity, hypercholesterolemia,
It may be used as a method for preventing / ameliorating hypertriglyceridemia, fatty liver, diabetes, hypertension, as well as chronic nephritis, nephrosis, cirrhosis, and obstructive jaundice. At that time, mammals in which suppression of apo B secretion is required include a tendency toward high cholesterol, a tendency toward high triglyceride, a tendency toward arteriosclerosis, a tendency toward fatty liver, a tendency toward hyperglycemia, and a tendency toward obesity based on the promotion of apo B secretion. Examples include humans, livestock, and companion animals, and humans, livestock, and companion animals having a lifestyle that promotes apoB secretion such as excessive intake of lipids, excessive intake of energy, and lack of exercise.

As a method for suppressing the secretion of apo B of the present invention, the lipid composition for suppressing apo B secretion of the present invention may be added to a target mammal as it is or as a food, drug,
What is necessary is just to carry out by ingesting as a feed. To be ingested means to be taken into the body orally or parenterally, whether spontaneously or other sources.

In this case, the amount to be taken into the mammal is 0.1
mg / kg / day or more, preferably 1 mg / kg / day or more, more preferably 10 mg / kg / day or more. If the amount is less than 0.1 mg / kg / day, the effect of suppressing apoB secretion may not be observed.

[0036]

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.

Example 1 Preparation of Plant Seed Oil 5 kg each of dried pomegranate seeds, calendula seeds, bitter melon seeds, catalpa seeds, or balsam apple seeds was crushed by a small oil press (uniaxial screw expeller V-01 type:
The oils and fats obtained were centrifuged at 10,000 G for 30 minutes to obtain 500 g of clear pomegranate seed oil and calendula seed oil 4.
50 g, bitter melon seed oil 400 g, catalpa seed oil 35
0 g and 250 g of balsam apple seed oil were obtained.

Example 2 Preparation of Conjugated Polyunsaturated Fatty Acid Having Conjugated Triene Structure Each of the pomegranate seed oil, calendula seed oil, bitter melon seed oil, catalpa seed oil, balsam apple seed oil prepared in Example 1 above, and After dissolving 10 g of commercially available tung oil in 10 volumes of ethanol, 1/10 volume of 33% KOH was added and
Hydrolysis was carried out at 0 ° C. for 45 minutes. 1 volume of water for decomposition products,
After adding twice the volume of hexane to extract and remove unsaponifiable matter, the aqueous phase was separated and adjusted to pH 2.5, extracted with n-hexane, and the solvent was distilled off under reduced pressure to obtain free fatty acids. Was. The obtained free fatty acid was subjected to silica gel column chromatography (carrier: Wakogel C-200, mobile phase: n-hexane (washing), n-hexane containing 20% by volume of ethanol (elution)) to obtain a purified fatty acid fraction. The yield was 8.8 g from pomegranate seed oil, 9.1 g from calendula seed oil,
9.0 g from bitter melon seed oil and 9.9 from catalpa seed oil.
0 g, 8.0 g from balsam apple seed oil, and 9.2 g from tung oil.

To analyze the fatty acid composition of each purified fatty acid fraction, each of the obtained fatty acid fractions was treated with a sulfuric acid-methanol solution (2: 230, volume ratio) and fatty acid methyl ester in the presence of dimethyl sulfoxide. After that, the product was analyzed by gas chromatography under the following conditions.

System: HP 5890 Series II Column: SPELUCO SP-2380, 100 mm
× 0.25 mm ID Column temperature: 185 ° C. Injection temperature: 200 ° C. Detector temperature: 210 ° C. The results of the analysis are summarized in Table 1.

[0041]

[Table 1] (Example 3) Effect of conjugated trienoic acid on apo B secretion by hepatocytes 10% fetal bovine serum (GIBUCO), penicillin (Nippon Shinyaku Co., Ltd., 100 units / ml), streptomycin (Nippon Shinyaku Co., Ltd.) , 0.1 mg / ml) containing DMEM medium (manufactured by Nissui Co., Ltd.)
After making human hepatocytes (HepG2) semi-confluent (7 × 10 ⁵ / well / ml) in a 4-well plate, the cells were washed twice with DMEM medium and DMEM containing 1% bovine serum albumin (manufactured by Sigma). After pre-incubation with the medium for 24 hours, α-linolenic acid (manufactured by Sigma, α-linolenic acid content 95%), the purified fatty acid fraction derived from pomegranate seed oil prepared in Example 2 (punicic acid content 71.
6%) or the purified fatty acid fraction derived from bitter melon seed oil prepared in Example 2 (α-eleostearic acid content 51.1)
%) Is α-linolenic acid, punicic acid, or α
-The medium was replaced with a DMEM medium containing 1% bovine serum albumin dissolved as eleostearic acid to a final concentration of 10 µM, and the cells were cultured for further 6 hours or 24 hours.
As a control, a DMEM medium containing 1% bovine serum albumin was used for the same culture. Thereafter, a culture solution was collected from each well, and the cells were washed twice with cooled PBS phosphate buffered saline, and then 1 ml of PBS was added to each well.
, And cells were collected using a rubber policeman, homogenized using a sonicator, and the amount of protein was determined to be Lowl.
y method (Lowry OHet. al., J. Biol. Chem.,
193, 265, 1951). On the other hand, H
Apo B secreted into the medium from the epG2 cells was quantified by sandwich ELISA (enzyme linked immunosorbent assay). That is, 37 ° C.
2 hours coating buffer (15 mM Na ₂ C
O ₂ , 31 mM NaHCO ₃ , pH 9.6) and anti-human I
After coating with a gG rabbit antibody (17.3 mg / L), Block Ace (0.5% NaN ₃ ) 0.1 ml /
The wells were further treated for 2 hours, and Tween-20 (0.
A 96-well plate washed three times with PBS containing 5 ml / L) was prepared, and the medium was added to each well of the plate with PBS.
A sample diluted 20-fold with the above and an LDL-apoB standard solution (0.5 to 20 mg / ml) were added, and after incubation at 4 ° C. for 16 hours, the plate was washed three times with PBS and then bound with horseradish peroxide. Anti-human Ig
G rabbit antibody was added, incubated at 37 ° C. for 2 hours, washed three times with PBS, phenylenediamine substrate solution (0.1 ml) was added, and after reaction at 25 ° C. for 30 minutes, 4.5.
The reaction was stopped by adding 0.05 ml of MH ₂ SO ₄ to each well.
1 at 482 nm was measured using a microplate reader (B
IO-Rad 550). The results are shown in Table 2.

[0042]

[Table 2] Punicic acid and α-eleostearic acid were found to induce less apoB secretion than α-linolenic acid, which is known to secrete less apoB.

Similarly, the fatty acid mainly composed of calendic acid derived from calendula seed oil, the fatty acid mainly composed of catalpinic acid derived from catalpa seed oil, and the punicic acid prepared in Example 1 were neutralized with NaOH. As a result of evaluating the punicic acid Na salt obtained as described above, the same tendency as that of punicic acid and α-eleostearic acid was recognized. (Example 4) Lipid metabolism improving effect of pomegranate seed oil 10-week-old female C57BL / 6J mouse (average weight 22
g) (Nippon Charles River Co., Ltd.) with soybean oil 1
5%, lard 15%, cornstarch 15%, sucrose 20% added high-fat / high-sugar fractionated purified feed (casein 25%, cornstarch 14.9%, sucrose 20%, soybean oil 15%, lard 15% , Cellulose powder 5%, AIN-93 mineral mixture 3.5%,
AIN-93 1% vitamin mixture, choline bitartrate 0.2
5%, tert-butylhydroquinone 0.006%, L-cystine 0.375% (total energy 21338 KJ / k
g) After obesity by rearing for 4 weeks (manufactured by Oriental Yeast Co., Ltd.), the animals were divided into 8 animals / group, and fat-removed standard formula feed for growth period (AIN-93G: Oriental Yeast Co., Ltd.) 2% soybean oil as an essential fatty acid source and 7% pomegranate seed oil prepared in Example 1 (pomegranate seed oil group)
Or, in a test feed to which 7% of a control fat (soybean oil group) was added,
They were bred for an additional 3 weeks under free-feeding conditions. The feed was replaced every two or three days with fresh ones and the food intake was recorded. 3
After a week, after feeding the last meal, fasting for 16 hours,
Under ether anesthesia, whole blood was collected using a heparinized syringe,
Centrifugation was performed at 3000 rpm for 15 minutes under cooling to obtain plasma. The total cholesterol of the obtained plasma was measured by cholesterol C test Wako (manufactured by Wako Pure Chemical Industries, Ltd.), and triglyceride was measured by triglyceride G test Wako (manufactured by Wako Pure Chemical Industries, Ltd.). The results are shown in Table 3. still,
The food consumption of each group was 3 g / day on average per individual, and no significant difference was observed between the groups.

[0044]

[Table 3] In the group fed the pomegranate seed oil, compared to the soybean oil group,
A tendency to decrease the total cholesterol and triglyceride concentrations in plasma was observed, and it was confirmed that lipid metabolism was improved as compared with the soybean oil group. (Example 5) Preventive effect of pomegranate seed oil on fatty liver 10-week-old female C57BL / 6J mice (average weight 22
g) (Charles Japan Nippon Charles River Co., Ltd.) were divided into 5 animals / group. Control group 1 was a purified high-fat, high-sugar diet with the same composition as in Example 4, and control group 2 was fat-free. Pomegranate seed oil group made up 8% of lard, a high-fat, high-sugar fractionated refined feed, with 10% soybean oil added to the standard mixed feed for growing season (AIN-93G: manufactured by Oriental Yeast Co., Ltd.) The feed was replaced with pomegranate seed oil prepared in Example 1 and bred for an additional 4 weeks under free-feeding conditions. Feed is 2 days or 3 days
They were replaced with fresh ones every day, and the food intake was recorded. After fasting for about 16 hours from the last day, the animals were exsanguinated under ether anesthesia and the liver was removed. After fixing the liver to formalin, a paraffin section was prepared by a conventional method, stained with hematoxylin and eosin, and pathological tissue search was performed with an optical microscope. The degree of liver fat degeneration was classified into five grades, and the results were evaluated based on the number of individuals for each grade. Table 4 shows the results. still,
The food consumption of each group was 3 g / day on average per individual, and no significant difference was observed between the groups.

[0045]

[Table 4] In a high-fat, high-sugar diet (control group 1), an image of fat accumulation in the liver and a fatty liver was observed, and no such image was observed in a normal diet (control group 2). In the test group to which pomegranate seed oil was added, an image in which fat accumulation in the liver was improved and normalized was observed.

Separately, a similar study was carried out using bitter melon seed oil, catalpa seed oil, balsam apple seed oil, snake guard seed oil, and western cherry seed oil. As a result, a fatty liver prevention effect similar to pomegranate seed oil was recognized. . (Example 6) Abnormal carbohydrate metabolism improvement effect of pomegranate seed oil Five-week-old female diabetic-onset KK-Ay mouse (average body weight 27.5 g) (obtained from CLEA Japan, Inc.) was preliminarily reared for one week. One group (control group) was divided into 5 animals / group, and one group (control group) was AIN-93G feed (Oriental Yeast Co., Ltd.) from which fat was removed (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), one group (troglidazone group) was a modified AIN-93G diet supplemented with 0.2% of the insulin sensitizer troglidazone, and another group (pomegranate seed oil group) was Pomegranate seed oil 8 prepared in Example 1 by 8% of soybean oil of modified AIN-93G feed
The feeds were replaced with free-feeding conditions for 4 weeks. The feed was replaced every two or three days with fresh ones and the food intake was recorded. Blood is collected from the tail vein every week,
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 was 5.3 g / day on average per individual, and 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, one week after the start of the test, the blood sugar level exceeded 400 mg / dl and diabetes occurred. Thereafter, the hyperglycemic state of the control group continued until 4 weeks. On the other hand, in the troglidazone group and the pomegranate seed oil group, the blood sugar level was maintained at 200 mg / dl or less throughout the test period and remained almost at the normal value. From these results, it was confirmed that pomegranate seed oil, which is one of the lipid compositions of the present invention, has a diabetes preventing / ameliorating effect. (Example 7) Production of pomegranate seed oil-containing margarine and pie Hardened soybean oil (mp40 ° C) 60%, palm oil 20%,
80 parts of a fat / oil composition comprising 20% of corn oil, 0.3 part of lecithin, 0.3 part of monoglyceride, 16 parts of water, salt 2
Parts, 5 parts of pomegranate seed oil prepared as in Example 1,
Add 0.01 parts of vitamin E as an antioxidant,
Then, the mixture was emulsified at 50 V for 15 minutes using a TK homomixer while blowing nitrogen gas, and then rapidly cooled and kneaded to 15 ° C. to obtain a sheet-like margarine. 1250 g of margarine, 1500 g of soft flour, 1000 strong flour thus obtained.
g, powdered milk 60 g, salt 45 g, sugar 15
An aqueous solution obtained by dissolving 00 g in 1250 ml of water was added, and the mixture was gently mixed to form a dough. The dough was spread in a sheet shape, and then formed by repeating trifolding five times. (Example 8) Preparation of capsule for oral use Pomegranate seed oil 40 m obtained in the same manner as in Example 1
g, lactose 200 mg, starch 70 mg, polyvinylpyrrolidone 5 mg, crystalline cellulose 35 mg,
3 Fill into 100mg of gelatin capsule and make the surface 40m
An oral capsule was prepared by coating with g of gelatin. Similarly, oral capsules using bitter melon seed oil, catalpa seed oil, and balsam apple seed oil were prepared. (Example 9) Preparation of oral tablet Pomegranate seed oil, calendula seed oil, bitter melon seed oil, catalpa seed oil, and balsam apple seed oil obtained by the method of Example 1 were uniformly mixed with 70 g of lactose and 30 g of corn starch for 5 g each. And 25 ml of a 10% hydroxypropylcellulose solution is added thereto, followed by stirring and granulation. After drying, this is sized, 2 g of magnesium stearate and 2 g of talc are added and mixed, and a tablet can be produced with a rotary tableting machine. (Example 10) Preparation of pet food 80 parts chicken surimi, 10 parts minced lean beef, 1 soy protein
0 parts, pomegranate seed oil, bitter melon seed oil, calendula seed oil, balsam seed oil, fatty acid derived from pomegranate seed oil, paulownia oil-derived fatty acid or calendula seed oil derived in Example 2 prepared by the method of Example 1 3 fatty acids each
1 part of chemical seasoning, 1 part of tocopherol, 0.5 part of calcium, 0.01 parts of vitamins, 3 parts of starch and 1 part of sorbitol are kneaded with a food cutter and filled into sheep intestine. After heating and cooking at 30 ° C. for 30 minutes, air-drying was performed at 50 ° C. to obtain dry sausage type pet food.

[0047]

Industrial Applicability The lipid composition of the present invention which suppresses apoB secretion is different from a lipid containing a saturated fatty acid or a monoenoic fatty acid even though it is a lipid, because the secretion of apoB is strongly suppressed. Atherosclerosis based on enhanced apoB secretion in mammals, obesity, hypercholesterolemia, hypertriglyceridemia, fatty liver, lifestyle-related diseases represented by diabetes or hypertension, coronary artery disease, brain It is expected to prevent and improve arterial disease, chronic nephritis, nephrosis, cirrhosis, obstructive jaundice, etc., and can be expected to be applied to foods, pharmaceuticals, and feeds.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61K 31/232 A61K 31/232 35/78 35/78 CTSH A61P 3/04 A61P 3/04 3 / 06 3/06 3/10 3/10 9/10 9/10 43/00 105 43/00 105 F term (reference) 2B150 AA01 AA06 AB03 AB10 DA55 DA58 DD45 4B018 MD10 MD15 ME03 ME04 4C088 AB12 AB19 AB26 AB46 AB52 BA18 CA03 NA05 ZA45 ZA70 ZC33 ZC35 4C206 AA01 AA02 DA05 DB09 DB43 DB47 MA01 MA04 NA05 ZA45 ZA70 ZC33 ZC35 4H059 BA26 BA30 BA33 BA34 BB05 BC13

Claims (12)

[Claims] An apolipoprotein B secretion inhibiting lipid composition comprising at least one member selected from the group consisting of conjugated trienoic acid-based fatty acids, salts thereof, and derivatives thereof. 2. Apolipoprotein B stronger than α-linolenic acid
The apolipoprotein B according to claim 1, wherein the apolipoprotein B contains at least one member selected from the group consisting of a conjugated trienoic acid-based fatty acid, a salt thereof, and a derivative thereof, which suppresses secretion.
Secretion inhibiting lipid composition. 3. The conjugated trienoic acid-based fatty acid is at least one selected from the group consisting of punicic acid, calendic acid, jarcaric acid, α-eleostearic acid, β-eleostearic acid, and catalpic acid. The lipid composition for inhibiting apolipoprotein B secretion according to claim 1 or 2, characterized in that: 4. The lipid composition for inhibiting apolipoprotein B secretion according to claim 1, wherein the derivative of the conjugated trienoic acid-based fatty acid is an ethyl ester derivative or a glycerol ester derivative of the conjugated trienoic acid-based fatty acid. . 5. The glycerol ester derivative of a conjugated trienoic acid fatty acid is selected from the group consisting of pomegranate seed oil, calendula seed oil, catalpa seed oil, bitter melon seed oil, balsam apple seed oil, abragiri seed oil, and cherry seed oil. The apolipoprotein B secretion inhibiting lipid composition according to any one of claims 1 to 4, which is at least one kind. 6. A food comprising the lipid composition for inhibiting apolipoprotein B secretion according to any one of claims 1 to 5. 7. A method for preventing or improving at least one member selected from the group consisting of atherosclerosis, obesity, hypercholesterolemia, hypertriglyceridemia, fatty liver, diabetes and hypertension. The food according to claim 6, wherein 8. A pharmaceutical comprising the lipid composition for inhibiting apolipoprotein B secretion according to any one of claims 1 to 5. 9. A method for preventing or ameliorating at least one selected from the group consisting of atherosclerosis, obesity, hypercholesterolemia, hypertriglyceridemia, fatty liver, diabetes, and hypertension. The pharmaceutical product according to claim 8, wherein 10. A feed comprising the apolipoprotein B secretion-inhibiting lipid composition according to any one of claims 1 to 5. 11. The lipid composition for inhibiting apolipoprotein B secretion according to any one of claims 1 to 5, and the lipid composition for inhibiting apolipoprotein B secretion according to any one of claims 1 to 5, for a mammal in which inhibition of apolipoprotein B secretion is required. A method for suppressing apolipoprotein B secretion, comprising ingesting at least one member selected from the group consisting of foods, medicines, and feeds comprising: 12. A lipid composition selected from the group consisting of the apolipoprotein B secretion-inhibiting lipid composition according to any one of claims 1 to 5, and a food, drug, or feed comprising the apolipoprotein B secretion-inhibiting lipid composition. Prevention and / or improvement of at least one selected from the group consisting of atherosclerosis, obesity, hypercholesterolemia, hypertriglyceridemia, fatty liver, diabetes or hypertension, characterized by ingesting at least one type. how to.