JP2023038107A - Lipid synthesis inhibitor, and food and drink, processed food for inhibited lipid synthesis and expression inhibitor comprising the same - Google Patents

Abstract

To provide a lipid synthesis inhibitor, and food and drink, and processed food for inhibited lipid synthesis comprising the same.SOLUTION: The present invention provides a lipid synthesis inhibitor, and food and drink, and processed food for inhibited lipid synthesis comprising the same, in which resorcinol of the formula (I) is comprised as an active ingredient. The resorcinol preferably includes extract extracted from wheat bran (where R1 is a C15 to 25 alkyl group or C15 to 25 alkenyl group, and R 2 is a hydrogen atom or a methyl group).SELECTED DRAWING: None

Description

TECHNICAL FIELD The present invention relates to a lipid synthesis inhibitor, a food or drink containing the same, a processed food for lipid synthesis inhibition, and a gene expression inhibitor for a protein involved in fatty acid synthesis.

Various diseases caused by excessive food intake, lack of exercise, stress, etc. increase the risk of arteriosclerosis and even cardiovascular diseases such as myocardial infarction and stroke. Hyperlipidemia, in which blood lipids such as triglycerides increase, is known as a risk factor for arteriosclerosis and ischemic heart disease. The synthesis process of triglycerides in blood is as follows. In vivo, acetyl-CoA carboxylase (hereinafter also referred to as "ACC") and fatty acid synthase Fatty acid are produced from acetyl Coenzyme A (CoA), which is produced mainly in the liver from sugar or the like as a starting material. Synthase (hereinafter also referred to as “FAS”), Elongation of long chain fatty acids 6 (hereinafter also referred to as “ELOVL6”) and the like are involved in biosynthesis of fatty acids. ACC, FAS, and ELOVL6 are all positively regulated by Sterol Regulatory Element Binding Transcription Factor 1 (hereinafter also referred to as "SREBF1"). (Non-Patent Documents 1 and 2). Synthesized fatty acids become triglycerides, which are secreted into the blood from the liver as lipoproteins.

Therefore, it is desired to develop a novel lipid synthesis inhibitor that inhibits triglyceride biosynthesis and is safe and sufficiently effective even when taken for a long period of time.

Furthermore, in recent years, it has been known that FAS is overexpressed in epithelial cancers of various tissues, and that inhibition of FAS and ACC may induce selective death of tumor cells (Non-Patent Document 3). Therefore, from the anticancer point of view as well, inhibitors that effectively suppress the expression of fatty acid synthase genes such as FAS and ACC are desired.

The present applicant has previously disclosed an anti-obesity agent (see Patent Document 1) containing, as an active ingredient, a peak component in partition chromatography of an alcohol extract of gramineous plant seeds, and an anti-obesity agent that maximizes the action and effect of the active ingredient. We proposed a method of ingesting the food obtained (see Patent Document 2). Also, a specific resorcinol lipid decomposition accelerator was proposed (Patent Document 3).

JP 2013-40108 A JP 2016-132641 A JP 2021-016363 A

Basic Aging Research 37(3); 2013, p29-31 Seikagaku Vol.80 No.8, August 2008, p762-767 Mashima, T., Seimiya, H., and Tsuruo, T. (2009). De novo fatty-acid synthesis and related pathways as molecular targets for cancer therapy. Br. J. Cancer 100, 1369-1372.

However, in Patent Documents 1 to 3, there is no particular study on the effect of suppressing the biosynthesis of lipids such as triglycerides and the effect of suppressing the expression of proteins involved in fatty acid synthesis such as FAS and ACC.

Accordingly, an object of the present invention is to provide a lipid synthesis inhibitor, a food or drink containing the same, a processed food for lipid synthesis inhibition, and a gene expression inhibitor for proteins involved in fatty acid synthesis.

As a result of intensive studies from the viewpoint of suppressing lipid synthesis in hepatocytes and the like, the present inventors newly discovered that the intake of specific resorcinol can suppress triglyceride biosynthesis in hepatocytes and the like. The present invention has been completed based on these findings.

（式中、Ｒ₁は炭素原子数１５～２５のアルキル基又は炭素原子数１５～２５のアルケニル基を表し、Ｒ₂は水素原子又はメチル基を表す。） That is, the present invention provides a lipid synthesis inhibitor containing resorcinol represented by the following formula (I) as an active ingredient.

(In the formula, R ₁ represents an alkyl group having 15 to 25 carbon atoms or an alkenyl group having 15 to 25 carbon atoms, and R ₂ represents a hydrogen atom or a methyl group.)

The present invention also provides food and drink containing the lipid synthesis inhibitor.

（式中、Ｒ₁は炭素原子数１５～２５のアルキル基又は炭素原子数１５～２５のアルケニル基を表し、Ｒ₂は水素原子又はメチル基を表す。） Furthermore, the present invention provides a processed food for inhibiting lipid synthesis, containing resorcinol represented by the following formula (I) as an active ingredient.

(In the formula, R ₁ represents an alkyl group having 15 to 25 carbon atoms or an alkenyl group having 15 to 25 carbon atoms, and R ₂ represents a hydrogen atom or a methyl group.)

Furthermore, the present invention provides an ACC expression inhibitor containing resorcinol represented by the above formula (I), an FAS expression inhibitor containing resorcinol represented by the above formula (I), and a FAS expression inhibitor represented by the above formula (I). and a resorcinol-containing SREBF1 expression inhibitor represented by the above formula (I).

According to the present invention, by administering or ingesting a lipid synthesis inhibitor, a food or drink containing the same, a processed food for lipogenesis inhibition, or a gene expression inhibitor for a protein involved in fatty acid synthesis, triglyceride production in hepatocytes or the like is induced. Synthesis can be effectively suppressed. Moreover, according to the present invention, gene expression of proteins involved in fatty acid synthesis, such as FAS, ACC, ELOVL6, and SREBF1, can be effectively suppressed.

FIG. 1(a) is a bar graph showing lipid accumulation in HepG2 cells of Examples and Comparative Examples. FIG. 2(a) is a bar graph showing the SREBF1 gene expression levels in Examples and Comparative Examples, and FIG. 2(b) is a graph showing the ACC gene expression levels in Examples and Comparative Examples. FIG. 3(a) is a bar graph showing FAS gene expression levels in Examples and Comparative Examples, and FIG. 3(b) is a graph showing sample distributions of ELOVL6 gene expression levels in Examples and Comparative Examples.

The present invention will be described below based on its preferred embodiments. The lipid synthesis inhibitor of this embodiment contains resorcinol represented by the following formula (I) as an active ingredient. This resorcinol is acetyl CoA carboxylase (ACC), fatty acid synthase Fatty acid synthase (FAS), Elongation of long chain fatty acids 6 (ELOVL6), genes related to proteins such as Sterol Regulatory Element Binding Transcription Factor 1 (SREBF1) for fatty acid synthesis. It suppresses the expression and has the effect of suppressing the synthesis of lipids such as triglycerides in hepatocytes and the like. In the following description, "X to Y" (X and Y are arbitrary numbers) means "X or more and Y or less" unless otherwise specified.
All of the following explanations about lipid synthesis inhibitors also apply to the ACC expression inhibitor, FAS expression inhibitor, ELOVL6 expression inhibitor, and SREBF1 expression inhibitor of the present invention.

（式中、Ｒ₁は炭素原子数１５～２５のアルキル基又は炭素原子数１５～２５のアルケニル基を表し、Ｒ₂は水素原子又はメチル基を表す。）

(In the formula, R ₁ represents an alkyl group having 15 to 25 carbon atoms or an alkenyl group having 15 to 25 carbon atoms, and R ₂ represents a hydrogen atom or a methyl group.)

Examples of the alkyl or alkenyl group having 15 to 25 carbon atoms represented by R ₁ in the above formula (I) include linear, branched or cyclic ones, and substituted or unsubstituted ones. R ₁ is attached to R ₂ at the meta or para position. Resorcinol having such functional groups and substitution positions may be used alone, or may be used as a mixture of multiple types in combination. From the viewpoint of suppressing, it is preferably used as a mixture in which multiple types are combined.

Preferred examples of the resorcinol represented by the formula (I) include those containing at least one resorcinol represented by the following (a1) to (f1). In the following description, the resorcinols shown in (a1) to (f1) below are also collectively referred to as "ARs".

(a1) Resorcinol in which R ₁ is an alkyl group having 15 carbon atoms (hereinafter also referred to as "AR15").
(b1) Resorcinol in which R ₁ is an alkyl group having 17 carbon atoms (hereinafter also referred to as "AR17").
(c1) Resorcinol in which R ₁ is an alkyl group having 19 carbon atoms (hereinafter also referred to as "AR19").
(d1) Resorcinol in which R ₁ is an alkyl group having 21 carbon atoms (hereinafter also referred to as "AR21").
(e1) Resorcinol in which R ₁ is an alkyl group having 23 carbon atoms (hereinafter also referred to as "AR23").
(f1) Resorcinol in which R ₁ is an alkyl group having 25 carbon atoms (hereinafter also referred to as "AR25").

Among these, the alkyl group represented by R ₁ is preferably linear or unsubstituted, more preferably linear, and still more preferably linear and unsubstituted. Examples of linear and unsubstituted alkyl groups include n-pentadecyl, n-heptadecyl, n-nonadecyl, n-henicosyl, n-tricosyl, n-pentacosyl and the like.

R ₂ in formula (I) above is preferably a hydrogen atom, and R ₁ is preferably bonded to R ₂ at the para-position.

From the viewpoint of effectively exhibiting the lipid synthesis inhibitory action, the combination of R ₁ and R ₂ is such that R ₁ is a linear and unsubstituted alkyl group, R ₂ is a hydrogen atom, and R ₁ is R ₂ It is more preferable to be bound in the para position with respect to. Specific examples of such resorcinol include compounds shown in (a2) to (f2) below. These may be used singly or as a mixture of a plurality of species. A mixture is preferred. (a2) to (f2) correspond to (a1) to (f1) described above, respectively.

(a2) 1,3-dihydroxy-5-n-pentadecylbenzene (C15:0) as AR15
(b2) 1,3-dihydroxy-5-n-heptadecylbenzene (C17:0) as AR17
(c2) as AR19, 1,3-dihydroxy-5-n-nonadecylbenzene (C19:0)
(d2) 1,3-dihydroxy-5-n-henicosylbenzene (C21:0) as AR21
(e2) 1,3-dihydroxy-5-n-tricosylbenzene (C23:0) as AR23
(f2) 1,3-dihydroxy-5-n-pentacosylbenzene (C25:0) as AR25

From the viewpoint of more effectively suppressing the biosynthesis of triglycerides by suppressing the gene expression of SREBF1, FAS, ACC, and ELOVL6, when the resorcinol represented by formula (I) is used as a mixture, the content of each AR is It is preferably within the following range. The content of ARs can be measured, for example, by the HPLC method described below.

The content of AR15 is preferably 0.1% by mass to 10.0% by mass, more preferably 0.1% by mass to 5.0% by mass, in the total content of all resorcinols represented by formula (I). , particularly preferably 0.5% by mass to 1.5% by mass.
The content of AR17 is preferably 1.0% by mass to 20.0% by mass, more preferably 5.0% by mass to 15.0% by mass, in the total content of all resorcinols represented by formula (I). , and particularly preferably 8.0% by mass to 12.0% by mass.
The content of AR19 is preferably 25.0% by mass to 40.0% by mass, more preferably 27.5% by mass to 37.5% by mass, in the total content of all resorcinols represented by formula (I). , and particularly preferably 30.0% by mass to 35.0% by mass.
The content of AR21 is preferably 40.0% by mass to 55.0% by mass, more preferably 42.5% by mass to 52.5% by mass, in the total content of all resorcinols represented by formula (I). , and particularly preferably 45.0% by mass to 50.0% by mass.
The content of AR23 is preferably 1.0% by mass to 15.0% by mass, more preferably 2.5% by mass to 12.5% by mass, in the total content of all resorcinols represented by formula (I). , and particularly preferably 5.0% by mass to 10.0% by mass.
The content of AR25 is preferably 0% by mass to 5.0% by mass, more preferably 0% by mass to 2.0% by mass, particularly preferably It is 0% by mass to 1.5% by mass. That is, AR25 may be substantially free.

The mixture of resorcinols represented by formula (I) described above may contain one or more resorcinols other than ARs. Other resorcinols include, for example, resorcinol in which R ₁ in formula (I) is an alkyl group having 27 carbon atoms, and particularly preferably R ₁ is a straight-chain unsubstituted alkyl group, and R A compound in which ₂ is a hydrogen atom and R ₁ is bonded to R ₂ in the para position. Examples of such compounds include 1,3-dihydroxy-5-n-heptacosylbenzene (C27:0).

Resorcinol represented by formula (I) can be synthesized by a conventional method, and can also be obtained as a commercial product. An extract extracted from a plant by a conventional method can also be used. When extracts from plants are used, plants containing resorcinol represented by formula (I) include Anacardiaceae, Ginkgoaceae, Proteaceae, Myrtaceae, Primulaceae, Myrtleaceae, Iridaceae, Araceae, and chrysanthemums. Mugwort, Legumes, Poaceae, etc. of the family. Among these, plants of the family Poaceae or Anacardiaceae are preferred as the source from the viewpoints of being edible, having a high content of resorcinol represented by formula (I), and being excellent in extraction efficiency. That is, the lipid synthesis inhibitor preferably contains, as resorcinol represented by formula (I), a resorcinol-containing extract represented by formula (I) extracted from a plant of the family Poaceae or a plant of the Anacardiaceae family. The use of a plant extract is particularly advantageous in that it can be obtained more easily as a mixture containing the above-mentioned ARs.

Examples of gramineous plants that can be used as a source of resorcinol represented by formula (I) include wheat, durum wheat, rye, triticale, barley, oats, potato, corn, rice, barnyard millet, millet, Millet and the like can be mentioned, and these can be used alone or in combination of two or more. The gramineous plant used for extraction may be seeds, pulverized, powdered or dried products obtained by cutting, pulverizing, or drying the seeds, or may use only the hulls, or may be used in combination. There may be. Specific examples including the outer skin include bran, powder, rice husk, rice bran, and the like.

Plants of Anacardiaceae that can be used as a source of resorcinol represented by formula (I) include, but are not limited to, nuts such as cashews. The Anacardiaceae plant used for extraction may be in any form as described above. For example, the plant or seed may be used as it is, or it may be cut, pulverized, dried, or otherwise pulverized, powdered, or dried. may be used, or a combination of these may be used.

Among these, from the viewpoint of efficiently obtaining an extract having a high lipid synthesis inhibitory activity, an extract obtained by extracting from a plant belonging to the genus Triticum such as wheat and durum wheat, or a plant belonging to the genus Ryme such as rye as a gramineous plant. It is more preferable that it is an extract extracted from wheat. Moreover, from the same point of view, it is preferably an extract obtained by extracting cashew nuts as a plant belonging to the Anacardiaceae family.
In particular, an extract extracted from wheat bran is preferable because it suppresses the gene expression of SREBF1, FAS, ACC, and ELOVL6, thereby suppressing triglyceride biosynthesis.

When an extract containing resorcinol represented by formula (I) is obtained from a plant, the extraction method is not particularly limited, but it is preferably obtained by alcohol extraction from the viewpoint of improving the extraction rate and convenience. . The extraction method using alcohol includes, for example, a method of immersing, stirring, or refluxing the seeds of various forms of the Poaceae plant or Anacardiaceae plant in alcohol, as well as a supercritical fluid extraction method and the like. In the case of the former method, the extraction temperature is preferably 2° C. to 100° C., the extraction time is preferably 30 minutes to 72 hours, and the amount of alcohol used is 50 parts by weight per 100 parts by weight of the seeds of the gramineous or Anacardiaceous plant. ~2000 parts by mass is preferred.

Examples of alcohols used for extraction include alcohols that are liquid at room temperature (25° C.). Specifically, lower monohydric alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol, 1, Polyhydric alcohols such as 3-butylene glycol, propylene glycol and glycerin, preferably those having 1 to 4 carbon atoms. Among these alcohols, it is preferable to use ethanol from the viewpoint of operability and environmental friendliness. As the alcohol used for extraction, hydrous ethanol containing aqueous components other than alcohol (water, pure water, distilled water, tap water, acidic water, alkaline water, neutral water, etc.) can also be used. The alcohol content in the hydrous alcohol is usually 70% by volume or more, preferably 80% by volume or more, more preferably 90% by volume or more.

The extract containing resorcinol represented by formula (I) may be used as it is, or the extract may be further purified. From the viewpoint of effectively expressing the lipid synthesis inhibitory action of resorcinol represented by formula (I), it is preferable to use a purified product obtained by purifying the extract. The purification method is not particularly limited as long as it is a method that yields the active ingredient of the lipid synthesis inhibitor of the present invention (resorcinol represented by formula (I) or a mixture thereof), but partition chromatography may be used. is preferable from the viewpoint of simplicity. A specific example of the partition chromatography method is preferably a normal phase chromatography method using a non-aqueous solvent as a mobile phase, and a known method such as an open column method, a medium pressure column method, or high performance liquid chromatography is appropriately selected. be able to.

Mobile phases in partition chromatography include lower monohydric alcohols having 1 to 4 carbon atoms such as methanol, ethanol, n-propanol, isopropanol, n-butanol, and 1,3-butylene glycol, propylene glycol, glycerin and the like. alcohols that are liquid at room temperature (25° C.) such as polyhydric alcohols having 1 to 4 carbon atoms; ethers such as diethyl ether and propyl ether; esters such as butyl acetate and ethyl acetate; ketones such as acetone and ethyl methyl ketone; hexane; methylene chloride; acetonitrile; and chloroform. When a plurality of solvents are combined to form a mobile phase, an isocratic mode in which the mixing ratio of the plurality of solvents is kept constant during partition chromatography, or a gradient mode in which the mixing ratio is changed may be used.

Any carrier capable of supporting and releasing the desired active ingredient can be used as the carrier in partition chromatography, and generally silica gel, polyacrylamide gel, dextran gel and the like can be mentioned.

The detection wavelength in partition chromatography may be from 170 to 320 nm, preferably from 200 to 300 nm.

Examples of partition chromatography suitable for purifying the alcoholic extract of Poaceae seeds or Anacardiaceae seeds include the partition chromatography method described in the Examples below. By using this method, a purified product containing resorcinol represented by formula (I) having a lipid synthesis inhibitory action can be obtained efficiently. The resulting purified product can be resorcinol consisting of a single compound or a mixture of resorcinols.

The content of resorcinol represented by formula (I) with respect to the total mass of the lipid synthesis inhibitor is preferably 70% by mass or more, more preferably 75% by mass, as the total content of resorcinol represented by formula (I). As mentioned above, it is particularly preferably 100% by mass, and the lipid synthesis inhibitor may be composed only of resorcinol represented by formula (I).

The lipid synthesis inhibitor is an extract, mixture or purified product containing resorcinol represented by formula (I), and if necessary, various pharmaceutically acceptable carriers, excipients, and other additives. A composition containing one or more of the agent and other components. The lipid synthesis inhibitor can be formulated by a conventional method, and the dosage form is preferably an oral preparation such as tablets, powders, granules, capsules, liquid preparations, injections, or transdermal preparations. can be done. In addition, other ingredients include ingredients having medicinal effects, such as anti-inflammatory drugs, various vitamins, herbal medicines, and minerals.

The lipid synthesis inhibitor may be directly administered or ingested to humans or animals for non-medical purposes, or the lipid synthesis inhibitor may be incorporated in food, drink or animal feed for ingestion. Animals include dogs, cats, mice, rats, hamsters, rabbits, cows, horses, monkeys, and the like. That is, lipid synthesis inhibitors can be applied not only to humans but also to pets (pet animals), domestic animals, etc., and can be applied to mammals in general for non-medical purposes.

The content of resorcinol represented by formula (I) in the lipid synthesis inhibitor can be changed as appropriate depending on the dosage form of the lipid synthesis inhibitor or the symptoms, age, sex, etc. of the person administering or taking it. Conventionally, in the case of human subjects, the content of resorcinol is known to be 0.01 g to 10 g per day per adult (converted to 60 kg), and the same amount can be used in this application. . This amount can be the total resorcinol content of formula (I). The method of administration or ingestion can be one-time or multiple-time administration, bolus administration, or continuous administration.

The lipid synthesis inhibitor is advantageous in that the convenience of administration or intake is increased by orally administering or ingesting it, such as by including it in food or drink. In the case of food and drink containing a lipid synthesis inhibitor, for example, staple foods such as bread and noodles, jellied foods and various snacks, baked goods, cakes, chocolate, gum, candy, tablets and other confectionery, capsules, Foods such as sauces, soups, dairy products, frozen foods, instant foods, processed foods such as supplements, drinking water, tea beverages, coffee beverages, milk beverages, fruit juice beverages, carbonated beverages, alcoholic beverages, soft drinks, nutrition Examples include, but are not limited to, beverages such as drinks, or edible seasonings. The manner in which the lipid synthesis inhibitor is contained in the food or drink is not particularly limited. may be included. That is, the food, drink and processed food of the present invention contain resorcinol represented by the formula (I), and are preferably used for suppressing lipid synthesis. In this case, the content of resorcinol represented by formula (I) is preferably contained within the above range.

In addition, there are no particular restrictions on the manner in which the lipid synthesis inhibitor is added to the animal feed. It may be included or mixed or dispersed in the animal drinking water. That is, the animal feed contains resorcinol represented by the formula (I), and is preferably used for suppressing lipid synthesis.

When administering or ingesting resorcinol represented by formula (I) to any animal, administration or ingestion can be carried out once or in multiple doses, or by bolus administration. It can be done continuously.
The following doses are conventionally known for resorcinol, and similar doses can be employed for this application. As the total content of resorcinol represented by formula (I), it is preferable to administer or ingest the following amounts.
・For dogs (weight equivalent to 10 kg): 200 μg to 500 mg per day
・For cats (weight equivalent to 3 kg): 200 μg to 500 mg per day
・For mice (weight equivalent to 30 g): 120 μg to 200 mg per day
・For hamsters (weight equivalent to 30 g): 120 μg to 200 mg per day
・For rabbits (weight equivalent to 2 kg): 200 μg to 500 mg per day

The intake period of the above-mentioned lipid synthesis inhibitor, food and drink, processed food, or animal feed is not particularly limited, but it is preferable to take it continuously for a certain period of time. Continuous ingestion for a week or more, preferably 3 weeks or more, particularly 7 weeks or more is preferred.

The present invention includes a method of administering or ingesting resorcinol represented by formula (I) or a mixture thereof to suppress biosynthesis of lipids such as triglycerides in hepatocytes (liver) and the like. As shown in the examples below, resorcinol used in the present invention suppresses the expression of enzymes that catalyze the synthesis of fatty acids such as ACC, FAS, and ELOVL6 in hepatocytes and the like, and inhibits the synthesis of fatty acids such as ACC, FAS, and ELOVL6. It suppresses the expression of SREBF1, which enhances the expression of the enzyme that catalyzes it. Therefore, resorcinol used in the present invention can effectively suppress the biosynthesis of triglycerides in hepatocytes and the like, and is effective in treating hyperlipidemia such as hypertriglyceridemia, fatty liver, arteriosclerosis and acute pancreatitis associated with hyperlipidemia. etc. can be improved. The present invention has discovered the possibility of improving hyperlipidemia and the like through pathways and mechanisms of action other than suppression of fat accumulation in adipocytes. In the present specification, simply referring to SREBF1 includes both SREBP1a and SREBP1c, which are transcription mutants.

Furthermore, the agent for suppressing gene expression of FAS, ACC, etc. of the present invention may contribute to the prevention or improvement of liver cancer such as hepatic epithelial cancer and various other cancers through suppression of gene expression of FAS, ACC, etc. be.

The method of suppressing lipid synthesis may involve administering or ingesting the above-described dosage and administration to, for example, humans and animals. The above doses may or may not be taken within 4 hours after awakening from sleep. The term "sleep" as used in the present invention is a natural state in which the person is unconscious but easily awakened, accompanied by a decrease in reaction to surrounding stimuli. Among them, only the one in which the state continued for the longest duration corresponds to "sleep." That is, sleep occurs only once in a 24-hour day, and, for example, apart from the several hours of sleep (sleep) that people normally take at night, they take a so-called nap for a relatively short period of time during the day. is not sleep as used herein. In addition, in the method of promoting suppression of lipid synthesis by administering or ingesting resorcinol represented by formula (I) or a mixture thereof, sleep time is not particularly limited, but is preferably 3 hours or more, more preferably 4 to 10 hours. It's time.

The present invention also encompasses products such as the aforementioned lipid synthesis inhibitors, food and drink, processed foods, or animal feeds, and commercial packages containing instructions. The instruction contains a statement that resorcinol represented by formula (I) is contained, a statement that the synthesis of lipids such as fat is suppressed, or a statement that the gene expression of FAS, ACC, EVOL6 or SREBF1 is suppressed, or It states that it inhibits fatty acid synthesis and at least one kind of usage and dosage, or at least one of these indications is attached. The form of this commercial package is not particularly limited, for example, a form in which an instruction is attached to a packaging container containing the product, a form in which an instruction is enclosed together with the product in the packaging container, and a packaging container itself. (a form in which the packaging container is the instruction manual).

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

[Production Example 1]
A resorcinol-containing extract was obtained from wheat bran as the seed of a gramineous plant by performing the following <extraction and purification method 1>, and the extract was purified to obtain a lipid synthesis inhibitor. The resorcinol represented by formula (I) contained as an active ingredient in the lipid synthesis inhibitor was a mixture of ARs having the following composition.
· 1,3-dihydroxy-5-n-pentadecylbenzene (C15:0): 1.2% by mass.
· 1,3-dihydroxy-5-n-heptadecylbenzene (C17:0): 10.9% by mass.
· 1,3-dihydroxy-5-n-nonadecylbenzene (C19:0): 33.9% by mass.
· 1,3-dihydroxy-5-n-henicosylbenzene (C21:0): 46.4% by mass.
· 1,3-dihydroxy-5-n-tricosylbenzene (C23:0): 7.5% by mass.
· 1,3-dihydroxy-5-n-pentacosylbenzene (C25:0): 0.1% by mass.

<Extraction purification method 1>
Five times the mass of ethanol was added to the wheat bran, and the mixture was extracted with stirring at 600 rpm and room temperature for 16 hours. This extraction dispersion was filtered to remove unnecessary substances, and an ethanol extract was recovered. Ethanol was distilled off from the extract to obtain an ethanol extract of wheat bran. Next, 1.5 times the amount (in terms of volume) of hexane was added to this ethanol extract to disperse it, and this dispersion was purified by medium pressure chromatography. Medium pressure chromatography conditions are as follows. A peak component appearing 31 to 36 minutes after the start of elution was collected and the solvent was distilled off to obtain the desired resorcinol mixture.

(Conditions for medium pressure chromatography)
・Column: silica gel (inject column 3 L, high flash column 5 L, 60 Å, 40 μm, manufactured by Yamazen Co., Ltd.)
・Mobile phase: in gradient mode, hexane/ethyl acetate mixed solvent (volume ratio) = 9 minutes at 90/10, 15 minutes at 80/20, 16 minutes at 60/40 ・Detection wavelength: 280 nm

<Method for quantifying resorcinol>
In this method, the resorcinol mixture obtained in <Extraction purification method 1> described above is quantified by high performance liquid chromatography (HPLC). Details are given below.
Methanol was added to the resorcinol mixture obtained in <Extraction purification method 1> so as to give a concentration of 200 μg/mL to prepare a methanol-added solution. This methanol-added solution was passed through a filter with a pore size of 0.45 μm, and the portion that passed through was subjected to high performance liquid chromatography (HPLC) analysis under the following conditions.

(HPLC conditions)
・ Column: Silica gel (ODS-80A, 5 μm, 4.6 × 250 mm, manufactured by GL Sciences Inc.)
・Guard column: ODS-80A, 5 μm, 4.6×50 mm,
・Column temperature: 30°C
・Mobile phase: 100% methanol
・Detection wavelength: 280 nm

[Examples and Comparative Examples]
Human liver cancer-derived cell line HepG2 (1×10 ⁵ cells/mL) was seeded in a 12-well plate in 10% by mass FBS-DMEM medium and cultured at 37° C. for 24 hours in a steam-saturated 5 vol% CO ₂ atmosphere. Thereafter, the medium was changed to FBS-DMEM medium containing no resorcinol (Comparative Example 1) and resorcinol represented by the formula (I) at a final concentration of 10 μM (Example 1). After culturing at 37° C. for 24 hours in a steam-saturated 5 vol % CO ₂ atmosphere, the amount of intracellular lipid accumulation in Comparative Example 1 and Example 1 was measured. The amount of intracellular lipid accumulation was evaluated by the Oil red O staining method. Furthermore, RNA was extracted from cells treated in the same manner and subjected to gene expression analysis. Comparative Example 1 and Example 1 were both carried out with n=3, and the measurement results of lipid accumulation were shown in FIG. 1 as mean±standard error (Mean±SE, n=3). Student's t-test was used for the test of significance, and p<0.05 was regarded as significant.

<Oil red O staining method>
After removing the medium and washing with PBS, 500 μL/well of 10 mass % formaldehyde was added to fix the cells. After washing with PBS, 500 μL/well of Oil red O staining solution was added and allowed to stand for 30 minutes. After removing the staining solution and washing with PBS, 1 mL/well of isopropanol was added and allowed to stand for 10 minutes. The suspended isopropanol-added solution was collected and the absorbance at OD 510 nm was measured on an absorbance plate.

(gene expression analysis)
Gene expression analysis of Example 1 and Comparative Example 1 was performed by the following procedure.
<RNA extraction>
Using the collected cells, RNA was extracted using TRI REAGENT (manufactured by Cosmo Bio).

<Reverse transcription reaction>
Using the extracted RNA, cDNA was synthesized using Prime Script RT reagent kit (manufactured by Takara). cDNA was synthesized using a thermal cycler under the conditions shown in Table 1 below.

<Gene expression analysis>
Using the obtained cDNA, gene expression analysis was performed by the SYBR Green method using CFX96 ^™ Real-Time System (manufactured by BIO RAD). SsoAdvance SYBR Green Supermix (manufactured by BIO-RAD) was used as a reagent. Analysis conditions and sequences of primers used are shown in Tables 3 to 5 below. The target genes were sterol regulatory element-binding transcription factor 1 (SREBF1c), acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), and elongation of long chain fatty acids L6 (EL6). Also, β-actin (ACBT) was used as a housekeeping gene.

The analysis was performed by creating a calibration curve based on the SQ values of serially diluted samples. Based on the ACBT and the SQ value of each target gene, the gene expression level of each sample was calculated from the following formula (1), and the relative ratio of the expression level when the average value of the expression level in Comparative Example 1 was 1 was calculated. Calculated. The results are shown in FIGS. 2(a) and (b) and FIGS. 3(a) and (b) as mean±standard error (Mean±SE, n=4). Student's t-test was performed between Comparative Example 1 and Example 1, and p<0.05 was considered significant.
Formula (1): Gene expression level of each sample = ([SQ value of target gene]/[SQ value of ACBT])

(result)
As shown in FIG. 1, in Example 1 to which resorcinol represented by formula (I) was added, the amount of lipid accumulation was significantly lower than in Comparative Example 1 to which no resorcinol was added. From these results, it was confirmed that resorcinol represented by formula (I) reduces the amount of lipid accumulation.
In order to examine in detail the mechanism of reducing the intracellular lipid accumulation amount in Example 1, the gene expression level was evaluated, and the results are shown in FIGS. Thus, in Example 1 to which resorcinol represented by formula (I) of Production Example 1 was added, the expression levels of each gene of SREBF1 (SREBF1c), ACC, FAS, and ELOVL6 were higher than in Comparative Example 1 to which no addition was added. significantly lower. As described above, it has been reported that these four genes are involved in lipid synthesis. Therefore, the reduction in lipid accumulation due to resorcinol represented by formula (I) in Production Example 1 is associated with lipid synthesis. It was confirmed that this was due to decreased expression of the involved genes.
As described above, according to the lipid synthesis inhibitor of the present invention, the food and drink containing the same, the processed food for lipid synthesis inhibition, and the gene expression inhibitor for proteins involved in fatty acid synthesis, by ingesting these, hepatocytes, etc. can effectively suppress the biosynthesis of lipids such as triglycerides in

Claims (7)

A lipid synthesis inhibitor containing resorcinol represented by the following formula (I) as an active ingredient.

(In the formula, R ₁ represents an alkyl group having 15 to 25 carbon atoms or an alkenyl group having 15 to 25 carbon atoms, and R ₂ represents a hydrogen atom or a methyl group.) 2. The lipid synthesis inhibitor according to claim 1, wherein the resorcinol is an extract extracted from a plant of the family Poaceae or a plant of the Anacardiaceae family. 2. The lipid synthesis inhibitor according to claim 1, which contains an extract extracted from wheat bran as said resorcinol. In the total mass content of resorcinol represented by formula (I),
0.1% by mass to 10.0% by mass of 1,3-dihydroxy-5-n-pentadecylbenzene,
1.0% by mass to 20.0% by mass of 1,3-dihydroxy-5-n-heptadecylbenzene,
25.0% by mass to 40.0% by mass of 1,3-dihydroxy-5-n-nonadecylbenzene,
40.0% by mass to 55.0% by mass of 1,3-dihydroxy-5-n-henicosylbenzene,
1.0% by mass to 15.0% by mass of 1,3-dihydroxy-5-n-tricosylbenzene,
4. Any one of claims 1 to 3, wherein the 1,3-dihydroxy-5-n-pentacosylbenzene is 0% by mass to 5.0% by mass and is used to suppress triglyceride biosynthesis in the liver. The lipid synthesis inhibitor according to the item. A food or drink containing the lipid synthesis inhibitor according to any one of claims 1 to 4. A processed food for suppressing lipid synthesis, containing resorcinol represented by the following formula (I) as an active ingredient.

(In the formula, R ₁ represents an alkyl group having 15 to 25 carbon atoms or an alkenyl group having 15 to 25 carbon atoms, and R ₂ represents a hydrogen atom or a methyl group.) at least one expression-suppressing factor selected from fat acid synthase, acetyl-CoA carboxylase A, elongation of long chain fatty acids 6 and sterol regulatory element binding transcription factor 1 containing resorcinol represented by the following formula (I) as an active ingredient agent.

(In the formula, R ₁ represents an alkyl group having 15 to 25 carbon atoms or an alkenyl group having 15 to 25 carbon atoms, and R ₂ represents a hydrogen atom or a methyl group.)

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