JP7266384B2 - Composition for suppressing migration of vascular smooth muscle cells containing n-hexane extract of cheese as an active ingredient - Google Patents

Description

The present invention relates to a composition for inhibiting migration of vascular smooth muscle cells.

In the past, arteriosclerosis progression inhibitory compositions containing lycopene, reduced-cholesterol cheese, and Lactobacillus plantarum Inducia strain DSM21379 as active ingredients have been reported (for example, JP-A-2009-505952 and JP-A-2012-524533). See publications, Japanese translation of PCT publication No. 2017-515799, etc.).

Japanese Patent Publication No. 2009-505952 Japanese Patent Publication No. 2012-524533 Japanese translation of PCT publication No. 2017-515799

By the way, the inventors of the present application have found that, as a result of long-term research, ingredients other than the above-mentioned active ingredients have an arteriosclerosis progression inhibitory effect. An object of the present invention is to provide a new active ingredient discovered by the inventors of the present application, and to provide an arteriosclerosis progression-suppressing composition containing the same active ingredient.

A composition for suppressing progression of arteriosclerosis according to one aspect of the present invention contains an n-hexane (normal-hexane) extract of cheese as an active ingredient. The "arteriosclerosis" referred to here is atherosclerosis. In addition, the "composition" referred to here includes formulations such as pharmaceuticals, supplements and food additives, food and drink (excluding animals and plants themselves), and animal products such as food and drink compositions (including processed food and drink). It includes things that can be ingested by humans (including humans).

By the way, when mouse aortic endothelial cells were seeded in a medium containing the above n-hexane extract of cheese and a cell migration inhibition test was carried out, the n-hexane extract of cheese had an inhibitory effect on the migration of mouse aortic endothelial cells. was recognized. That is, this arteriosclerosis progression inhibitory composition can also be expressed as a "vascular smooth muscle cell migration inhibitory composition". By the way, it has been revealed in the past that the progression of arteriosclerosis is suppressed when the migration of vascular smooth muscle cells is suppressed. Therefore, the aforementioned n-hexane extract of cheese functions as an active ingredient that suppresses the progression of arteriosclerosis.

In the composition for inhibiting the progression of arteriosclerosis or the composition for inhibiting vascular smooth muscle cell migration, it is clear that the n-hexane extract of cheese inhibits the progression of arteriosclerosis or vascular smooth muscle cell migration in a dose-dependent manner. It has become. That is, the higher the dose of the n-hexane extract of cheese, the higher the inhibitory effect on progression of arteriosclerosis or the inhibitory effect on vascular smooth muscle cell migration.

Moreover, in the composition for inhibiting progression of arteriosclerosis or the composition for inhibiting migration of vascular smooth muscle cells, the cheese is preferably at least one kind of cheese selected from the group consisting of camembert cheese and gorgonzola cheese. Moreover, it is preferable that the camembert cheese is "Meiji Hokkaido Tokachi (registered trademark) camembert cheese" manufactured by Meiji Co., Ltd. Gorgonzola cheese is preferably "Gorgonzola Dolce DOP" manufactured by Biraghi (Piedmont, Italy). This is because the n-hexane extract of these cheeses exhibits a particularly effective inhibitory effect on progression of arteriosclerosis or vascular smooth muscle cell migration.

In addition, the above-mentioned invention includes "a method of using an n-hexane extract of cheese as an arteriosclerosis progression inhibitor or vascular smooth muscle cell migration inhibitor", "as an arteriosclerosis progression inhibitor or vascular smooth muscle cell migration inhibitor n-hexane extract of cheese for use in", "a method of orally administering an n-hexane extract of cheese to a human to suppress the progression of arteriosclerosis or the migration of vascular smooth muscle cells" can. From another point of view, it can also be expressed as "use of n-hexane extract of cheese for producing a composition for inhibiting progression of arteriosclerosis or migration of vascular smooth muscle cells".

Another aspect of the invention is an n-hexane extract of cheese.

As described above, the n-hexane extract of this cheese has been found to have an inhibitory effect on mouse aortic endothelial cell migration. That is, the n-hexane extract of this cheese exerts an inhibitory effect on the migration of vascular smooth muscle cells. By the way, it has been clarified that when the migration of vascular smooth muscle cells is suppressed, the progression of arteriosclerosis is suppressed. Therefore, the n-hexane extract of this cheese can function as an active ingredient that inhibits the progression of arteriosclerosis.

The cheese to be extracted with n-hexane is preferably at least one kind of cheese selected from the group consisting of camembert cheese and gorgonzola cheese. Moreover, it is preferable that the camembert cheese is "Meiji Hokkaido Tokachi (registered trademark) camembert cheese" manufactured by Meiji Co., Ltd. This is because the n-hexane extract of these cheeses exhibits a particularly effective arteriosclerosis progression inhibitory effect (action) or vascular smooth muscle cell migration inhibitory effect (action).

In addition, in the n-hexane extract of this cheese, the active ingredient for the arteriosclerosis progression inhibitory action or vascular smooth muscle cell migration inhibitory action is a compound represented by the following general formula (1).

In the above formula, R ₁ represents a linear saturated alkyl group or unsaturated alkyl group having 17 carbon atoms, and R ₂ represents a linear unsaturated alkyl group having 17 carbon atoms.

A composition for suppressing progression of arteriosclerosis according to another aspect of the present invention contains a compound represented by the following general formula (1) as an active ingredient. The "arteriosclerosis" referred to here is atherosclerosis. In addition, the "composition" referred to here includes formulations such as pharmaceuticals, supplements and food additives, food and drink (excluding animals and plants themselves), and animal products such as food and drink compositions (including processed food and drink). It includes things that can be ingested by humans (including humans).

In the above formula, R ₁ represents a linear saturated alkyl group or unsaturated alkyl group having 17 carbon atoms, and R ₂ represents a linear unsaturated alkyl group having 17 carbon atoms.

By the way, when mouse aortic endothelial cells were seeded in a medium containing the compound represented by the above-described general formula (1) and a cell migration inhibition test was performed, migration of mouse aortic endothelial cells to the compound represented by general formula (1) was inhibited. It was found to have an inhibitory effect. That is, this arteriosclerosis progression inhibitory composition can also be expressed as a "vascular smooth muscle cell migration inhibitory composition". By the way, it has been revealed in the past that the progression of arteriosclerosis is suppressed when the migration of vascular smooth muscle cells is suppressed. Therefore, the compound represented by the general formula (1) functions as an active ingredient that suppresses the progression of arteriosclerosis.

As a result of many years of research by the inventors of the present application, the above-mentioned n-hexane extract of cheese, the composition for inhibiting vascular smooth muscle cell migration, and the composition for inhibiting the progression of arteriosclerosis have been found to inhibit progression of arteriosclerosis or vascular smooth muscle cell migration. can be effectively suppressed. Therefore, if an animal such as a human ingests an n-hexane extract of cheese, a composition for inhibiting migration of vascular smooth muscle cells, or a composition for inhibiting the progression of arteriosclerosis from normal times, migration of vascular smooth muscle cells , and by extension, the progression of arteriosclerosis can be suppressed.

n-hexane extract of “Meiji Hokkaido Tokachi (registered trademark) camembert cheese” manufactured by Meiji Co., Ltd. according to Example 1 (plot of ■ (black square)), Biraghi (Piedmont, Italy) according to Example 2 n-hexane extract of "Gorgonzola Dolce DOP" (▲ (black triangle) plot), acetonitrile extract of "Meiji Hokkaido Tokachi (registered trademark) camembert cheese" manufactured by Meiji Co., Ltd. according to Comparative Example 1 ( ● (black circle) plot), and the acetonitrile extract of "Gorgonzola Dolce DOP" manufactured by Biraghi (Piedmont, Italy) according to Comparative Example 2 (◆ (black diamond) plot) medium concentration of mouse aorta FIG. 4 is a line graph diagram showing the effect on endothelial cell viability. It is a figure which shows the procedure of a cell migration test. The n-hexane extract of "Meiji Hokkaido Tokachi (registered trademark) Camembert cheese" manufactured by Meiji Co., Ltd. according to Example 3, and the "Gorgonzola Dolce DOP" manufactured by Biraghi (Piedmont, Italy) according to Example 4 ” is a bar graph showing the effect of the medium concentration of n-hexane extracts of A. on the migratory cell number of mouse aortic endothelial cells. In the figure, "positive control" indicates the results of Control Example 1, and "negative control" indicates the results of Control Example 2. Acetonitrile extract of "Meiji Hokkaido Tokachi (registered trademark) Camembert cheese" manufactured by Meiji Co., Ltd. according to Comparative Example 3, and Biraghi (Piedmont, Italy) according to Comparative Example 4 "Gorgonzola Dolce DOP" manufactured by FIG. 4 is a bar graph diagram showing the effect of medium concentration of acetonitrile extract on migratory cell number of mouse aortic endothelial cells. In the figure, "positive control" indicates the results of Control Example 1, and "negative control" indicates the results of Control Example 2.

Hereinafter, the present invention will be described in detail by showing embodiments of the present invention, but the present invention is not limited to the individual forms described below.

<n-hexane extract of cheese>
The cheese n-hexane (normal hexane) extract according to the embodiment of the present invention can suppress the migration of vascular smooth muscle cells, as shown in [Example] described below, is expected to suppress the progression of arteriosclerosis. That is, the n-hexane extract of cheese can be used as an active ingredient of a composition for inhibiting migration of vascular smooth muscle cells or a composition for inhibiting progression of arteriosclerosis. In addition, this inhibitory effect is dose-dependent, and the higher the dose, the higher the inhibitory effect.

Cheeses to be extracted with n-hexane include, for example, fresh type cheese, washed type cheese, hard type cheese, semi-hard type cheese, blue mold type cheese, white mold type cheese, chevre type cheese, processed cheese and the like.

Examples of fresh cheese include cottage cheese, cream cheese, feta cheese, Brillat-Savarin cheese, fromage blanche, mascarpone cheese, mozzarella cheese, ricotta cheese, and string cheese. Washed cheeses include, for example, Epoisse cheese, Chaume cheese, Taleggio cheese, Pont-L'Eveque cheese, Manstel cheese, Mont d'Or cheese, Rami du Chambertin cheese, Langres cheese, and Livarot cheese. Hard type cheeses include, for example, Abondance cheese, Edam cheese, Emmental cheese, Granavadano cheese, Gruyere cheese, Comte cheese, Splints cheese, Cheddar cheese, Parmigiano Reggiano cheese, Pecorino Romano cheese, Beaufort cheese, Mimolette Examples include cheese and raclette cheese. Examples of semi-hard cheeses include Cantal cheese, Gouda cheese, Samseau cheese, Saler cheese, Saint-Nectaire cheese, Fontina cheese, Marivo cheese, Morbier cheese, Raiole cheese, Reblochon de Savoie cheese, Manzel Bavette cheese, and the like. be done. Blue mold type cheeses include, for example, Cambozola cheese, Gorgonzola cheese, Stilton cheese, Bavaria blue cheese, Flume d'Ambert cheese, Bleu du Vercors Sasnage cheese, Bleu de course cheese, Bleu de - Examples include Gex cheese and Roquefort cheese. Mildew-type cheeses include, for example, camembert cheese, churomier cheese, chaurse cheese, neuchâtel cheese, barraca cheese, brie de meurin cheese, brie de mou cheese, saint-andre cheese and the like. Examples of chevre-type cheese include Valensee cheese, Crotin de Chavignol cheese, Sainte-Mole de Touraine cheese, Ser-sur-Cher cheese, Briny-Saint-Pierre cheese, Shabishou du Poiteau cheese, and the like. be done. Processed cheese is obtained by heating and processing one or more of the above natural cheeses.

Among the above cheeses, camembert cheese and gorgonzola cheese are particularly preferred. Among Camembert cheeses, “Meiji Hokkaido Tokachi (registered trademark) Camembert cheese” manufactured by Meiji Co., Ltd. is particularly preferable, and among Gorgonzola cheeses, “Gorgonzola Dolce DOP” manufactured by Biraghi (Piedmont, Italy) is particularly preferable.

ここで、上式（１）中、Ｒ_１は炭素数１７の直鎖状の飽和アルキル基または不飽和アルキル基を示し、Ｒ_２は炭素数１７の直鎖状の不飽和アルキル基を示す。 <Phosphatidylethanolamine>
The phosphatidylethanolamine according to the embodiment of the present invention can suppress the migration of vascular smooth muscle cells, which in turn suppresses the progression of arteriosclerosis, as shown in [Examples] described below. It is expected. That is, phosphatidylethanolamine can be used as an active ingredient of a vascular smooth muscle cell migration inhibitory composition or an arteriosclerosis progression inhibitory composition. In addition, this inhibitory effect is dose-dependent, and the higher the dose, the higher the inhibitory effect. By the way, phosphatidylethanolamine according to an embodiment of the present invention is a compound represented by the following general formula (1).

Here, in the above formula (1), R ₁ represents a linear saturated alkyl group or unsaturated alkyl group having 17 carbon atoms, and R ₂ represents a linear unsaturated alkyl group having 17 carbon atoms.

Phosphatidylethanolamines according to embodiments of the present invention more specifically have chemical structures represented by the following chemical structural formulas (2) to (6). That is, in the formula (2) below, the linolyl group corresponds to R ₁ in the above formula (1), and the α-linolenyl group corresponds to R ₂ in the above formula (1). In the formula (3) below, the linolyl group corresponds to R ₁ and R ₂ in the formula (1) above. In the formula (4) below, the oleyl group corresponds to R ₁ in the formula (1) above, and the linolyl group corresponds to R ₂ in the formula (1) above. In the formula (5) below, the oleyl group corresponds to R ₁ and R ₂ in the formula (1) above. In the formula (6) below, the stearyl group corresponds to R ₁ in the formula (1) above, and the oleyl group corresponds to R ₂ in the formula (1) above. By the way, the phosphatidylethanolamine according to the embodiment of the present invention preferably contains at least phosphatidylethanolamines of chemical structural formulas (4) to (6), and at least phosphatidylethanolamines of chemical structural formulas (3) to (6). It more preferably contains an amine, and more preferably contains at least phosphatidylethanolamines of chemical structures (2) to (6).

By the way, the phosphatidylethanolamine according to the embodiment of the present invention is contained in n-hexane (normal hexane) extract of cheese. This phosphatidylethanolamine can be detected by liquid chromatography. The inventors of the present application detected the above-described phosphatidylethanolamine from an n-hexane (normal hexane) extract of cheese using the following conditions.

・Column: Accucore-RPMS 2.1×100 mm 2.6 μm
・Column temperature: 60°C
- Mobile phase A: 0.1% acetic acid solution of 10 mM ammonium acetate / acetonitrile = 90/10 (volume ratio) - Mobile phase B: 0.02% acetic acid solution of 2 mM ammonium acetate / 2-propanol / acetonitrile = 2/88/10 (capacity ratio)
・Flow rate: 0.25 mL/min ・Injection volume: 2 μL
- Gradient conditions: as described in Table 1 (development time: 43 minutes)

<Ingestion subject of n-hexane extract of cheese, composition for inhibiting vascular smooth muscle cell migration or composition for inhibiting progression of arteriosclerosis, and method of ingestion>
The n-hexane extract of cheese, the composition for inhibiting migration of vascular smooth muscle cells, or the composition for inhibiting the progression of arteriosclerosis according to the embodiment of the present invention can be ingested by animals, particularly humans, who wish to inhibit the progression of arteriosclerosis. to demonstrate its function. It should be noted that the “human” referred to here may be people of a wide range of ages, from children to the elderly, regardless of age or sex. In addition, the n-hexane extract of this cheese, the composition for inhibiting migration of vascular smooth muscle cells, or the composition for inhibiting progression of arteriosclerosis can effectively exert its function by being ingested by animals, particularly humans, before the progression of arteriosclerosis. Demonstrate. The term "ingestion" as used herein means that there is no limitation on the route of intake as long as the substance enters the body of an animal such as a human, and can be achieved by any of the known methods of intake, such as oral intake, tube intake, and enteral intake. At this time, oral ingestion via the gastrointestinal tract and enteral ingestion are typically mentioned, but oral ingestion is preferable, and oral ingestion through eating and drinking is more preferable.

The concentration of the n-hexane extract of cheese in the composition for inhibiting vascular smooth muscle cell migration or the composition for inhibiting progression of arteriosclerosis according to the embodiment of the present invention is n - Cheese and its n-hexane extract are preferably included so that the hexane extract contains 1 μg/mL or more, and the cheese or its n-hexane extract contains 5 μg/mL or more - more preferably contains a hexane extract, more preferably contains cheese or its n-hexane extract such that the n-hexane extract of the cheese contains 10 μg/mL or more, the n-hexane of the cheese More preferably, the cheese and its n-hexane extract are contained so that the extract contains 20 μg / mL or more, and the cheese and its n- It is particularly preferred to include a hexane extract. As described above, the higher the content of the n-hexane extract of cheese in the composition for inhibiting vascular smooth muscle cell migration or the composition for inhibiting progression of arteriosclerosis, the higher the effect, but the upper limit is, for example, 1 mg. /mL. Moreover, at this time, the cheese may be pulverized or cut.

The mass per unit package of the n-hexane extract of cheese according to the embodiment of the present invention is not particularly limited, but from the viewpoint that the effect can be sufficiently obtained and it is easy to consume at one time, it is 1 g or more. It is preferably within the range of 500 g or less, more preferably within the range of 5 g or more and 200 g or less, further preferably within the range of 10 g or more and 150 g or less, and preferably within the range of 10 g or more and 100 g or less. Most preferred. Moreover, the above-mentioned unit packaging may be not only unit packaging per bag, box, or container, but may be unit packaging for one time contained therein, or may be unit packaging per day. In addition, it is also possible to package a quantity suitable for intake for a plurality of days, for example, one week, or to include a plurality of individual packages.

The mass per unit package of the composition for inhibiting vascular smooth muscle cell migration or the composition for inhibiting progression of arteriosclerosis according to the embodiment of the present invention is not particularly limited. From the viewpoint of being easy to cut, it is preferably within the range of 1 g or more and 500 g or less, more preferably within the range of 5 g or more and 200 g or less, further preferably within the range of 10 g or more and 150 g or less, and 10 g or more. Most preferably it is in the range of 100g or less. Moreover, the above-mentioned unit packaging may be not only unit packaging per bag, box, or container, but may be unit packaging for one time contained therein, or may be unit packaging per day. In addition, it is also possible to package a quantity suitable for intake for a plurality of days, for example, one week, or to include a plurality of individual packages.

The n-hexane extract of cheese, the composition for inhibiting vascular smooth muscle cell migration, or the composition for inhibiting progression of arteriosclerosis according to the embodiment of the present invention is maintained for 1 week or more, preferably 2 weeks or more, more preferably 4 weeks or more. , should be taken continuously. Since the n-hexane extract, the composition for inhibiting vascular smooth muscle cell migration, or the composition for inhibiting the progression of arteriosclerosis according to the embodiment of the present invention can be safely ingested, the period of ingestion is not particularly limited and can be used permanently. can continue. In addition, the n-hexane extract, the composition for inhibiting vascular smooth muscle cell migration, or the composition for inhibiting progression of arteriosclerosis may be taken continuously only for a part of the period, or may be taken intermittently for any period of time. may be ingested.

<Forms of cheese n-hexane extract, composition for inhibiting vascular smooth muscle cell migration, or composition for inhibiting progression of arteriosclerosis>
The n-hexane extract of cheese, the composition for inhibiting migration of vascular smooth muscle cells, or the composition for inhibiting progression of arteriosclerosis according to the embodiment of the present invention can be used as pharmaceuticals or foods and drinks. The drug or food or drink is useful in that it has an inhibitory effect on migration of vascular smooth muscle cells or an inhibitory effect on progression of arteriosclerosis. When the n-hexane extract of cheese, the composition for inhibiting vascular smooth muscle cell migration, or the composition for inhibiting progression of arteriosclerosis according to the embodiment of the present invention is used as a pharmaceutical product or food or drink, n- A hexane extract may be used, or a combination of n-hexane extracts of two or more cheeses may be used.

When the n-hexane extract of cheese, the composition for inhibiting vascular smooth muscle cell migration, or the composition for inhibiting the progression of arteriosclerosis according to the embodiment of the present invention is used as a pharmaceutical product or a food or drink, their conditions are not particularly limited. , paste, spray-dried, freeze-dried, vacuum-dried, drum-dried, liquid dispersed in a medium, diluted with a diluent, crushed by crushing the dried product with a mill, etc. can be

Furthermore, the n-hexane extract of cheese, the composition for inhibiting migration of vascular smooth muscle cells, or the composition for inhibiting progression of arteriosclerosis according to the embodiment of the present invention can be used as a food with health claims or a food for the sick (suppressing cancer cachexia). active foods, etc.). The food with health claims system was established to cover not only ordinary foods but also foods in the form of tablets, capsules, etc., based on domestic and international trends and consistency with the conventional food for specified health use system. . The system stipulates three types: food for specified health use (individual permission type), food with nutrient function claims (standard type), and food with function claims. The n-hexane extract of cheese, the composition for inhibiting vascular smooth muscle cell migration, or the composition for inhibiting the progression of arteriosclerosis according to the embodiment of the present invention is used as a food for special purposes such as a food for specified health use or a food with nutrient function claims. By administering to such animals, it becomes possible to suppress the migration of vascular smooth muscle cells or to suppress the progression of arteriosclerosis.

The n-hexane extract of cheese, the composition for inhibiting vascular smooth muscle cell migration, or the composition for inhibiting progression of arteriosclerosis according to the embodiment of the present invention, its use, efficacy, function, type of active ingredient, and functional ingredient It is preferable to display explanations such as the type and intake method. The “labeling” referred to here should be appropriate labeling for pharmaceuticals, quasi-drugs, foods with health claims, foods for specified health uses, foods with function claims, general foods, health supplements, health foods, and supplements. . In addition, the "display" here includes all displays for informing consumers of the above description. This display may be any display that allows one to recall or analogize the display content described above, and may include all displays regardless of the purpose of display, content of display, object or medium to be displayed, and the like. For example, displaying the above description on product packaging and containers, displaying or distributing the above description on advertisements, price lists, or transaction documents related to products, or disseminating information containing these contents electromagnetically (Internet, etc.) ) method.

When the product obtained by packaging the n-hexane extract of cheese, the composition for inhibiting vascular smooth muscle cell migration, or the composition for inhibiting the progression of arteriosclerosis according to the embodiment of the present invention is, for example, a food or drink, is preferably labeled with, for example, "inhibition of vascular smooth muscle cell migration" and "inhibition of progression of arteriosclerosis", and the above-mentioned indications such as "prevent hardening of blood vessels" and "maintain supple blood vessels". It is also preferable that an indication having a meaning equivalent to is attached.

It should be noted that the wordings used for the display as described above are not limited to the above examples, and wordings having the same meaning as those described above may be used. Such phrases include, for example, "suppressing the migration of vascular smooth muscle cells", "suppressing the progression of arteriosclerosis", "useful in suppressing the migration of vascular smooth muscle cells", or " Various phrases such as "useful for suppressing progression of arteriosclerosis" are acceptable.

When the n-hexane extract of cheese, the composition for inhibiting vascular smooth muscle cell migration, or the composition for inhibiting progression of arteriosclerosis according to the embodiment of the present invention is used as food or drink, the type of food or drink is not particularly limited. Food and drink include, for example, milk, processed milk, soft drinks, cheese, other dairy products, bread, biscuits, crackers, pizza crust, powdered milk, liquid diet, food for the sick, powdered milk for infants, pregnant women and nursing mothers It may be a food such as powdered milk for women, a nutritious food, or the like. In the production of such food and drink, the n-hexane extract of cheese, the composition for inhibiting vascular smooth muscle cell migration, or the composition for inhibiting progression of arteriosclerosis according to the embodiment of the present invention is used as it is, or other Usual production methods for food compositions can be used, such as mixing with food and drink or food ingredients. Also, the shape of the food and drink is not particularly limited, and any shape of food and drink that is commonly used may be used. For example, it may be in any form such as solid (including powder and granules), paste, liquid, and suspension, and is not limited to these. At this time, milk drinks, soft drinks, jelly drinks, tablets, and powdered foods are more preferable.

The composition for inhibiting migration of vascular smooth muscle cells or the composition for inhibiting progression of arteriosclerosis according to the embodiment of the present invention contains water, proteins, carbohydrates, lipids, vitamins, minerals, organic acids, organic bases, fruit juice, Ingredients contained in ordinary foods such as flavors, functional ingredients, and food additives can be added without problems. In the production of the above food and drink, as the protein source, for example, soy protein, milk protein, chicken egg protein, animal and plant proteins such as meat protein, proteins or protein-containing raw materials that are commonly used in food production, such as hydrolysates thereof, are used. can do. Examples of sugar sources include processed starch (texturin, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.), dietary fiber, and the like. Lipid sources include, for example, lard, fish oil, fractionated oils thereof, hydrogenated oils, transesterified oils, and other animal fats; palm oil, safflower oil, corn oil, rapeseed oil, coconut oil, fractionated oils thereof; , hydrogenated oils, and vegetable oils such as transesterified oils. Examples of vitamins include vitamin A, carotenes, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline. , folic acid and the like, and minerals include, for example, calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, selenium and the like. Organic acids include, for example, malic acid, citric acid, lactic acid, and tartaric acid. Examples of functional ingredients include oligosaccharides, glucosamine, collagen, ceramide, royal jelly, and polyphenols. Food additives include, for example, emulsifiers, stabilizers, thickeners, gelling agents, sweeteners, acidulants, preservatives, antioxidants, pH adjusters, coloring agents, flavoring agents, and the like. Various milk-derived components such as butter, milk minerals, cream, whey, non-protein nitrogen, sialic acid, phospholipids, and lactose are examples of components that can be suitably used in the production of the above food and drink. In addition, any component having an inhibitory effect on cancer cachexia may be added to the composition for inhibiting vascular smooth muscle cell migration or the composition for inhibiting progression of arteriosclerosis according to the embodiment of the present invention.

These components can be used in combination of two or more. The raw material may be any of natural products, processed natural products, synthetic products and/or foods containing these in large amounts.

The n-hexane extract of cheese, the composition for inhibiting vascular smooth muscle cell migration, or the composition for inhibiting the progression of arteriosclerosis according to the embodiment of the present invention can be used as a vascular smooth muscle cell migration-inhibiting drug or an arteriosclerosis progression-inhibiting drug. can also When producing such pharmaceuticals, the n-hexane extract of cheese, the composition for inhibiting migration of vascular smooth muscle cells, or the composition for inhibiting progression of arteriosclerosis can be used as crushed or unpulverized processed products. In addition, the n-hexane extract of cheese used at this time may be a single n-hexane extract of cheese, or a combination of n-hexane extracts of two or more types of cheese. good too.

The content of the n-hexane extract of cheese in the drug can be arbitrarily determined according to its purpose and application. An example content is 150 μg/mL, but the embodiment of the present invention is not limited to this.

The dosage of the drug containing the above-mentioned n-hexane extract of cheese as an active ingredient can be appropriately set in consideration of various factors such as the route of administration, the age, body weight and symptoms of the target animal including humans. can. An example of a suitable dose is 0.1 g to 1000 g/kg/day of the active ingredient, but the embodiment of the present invention is not limited to this. Since the n-hexane extract of cheese, which is the active ingredient of the present invention, has no safety problems, it may be used in an amount larger than the above range.

The dosage form of the pharmaceutical product is preferably a dosage form that allows oral administration in order to allow the n-hexane extract of cheese to reach the intestine. Examples of preferred dosage forms of pharmaceuticals according to embodiments of the present invention include tablets, pills, coated tablets, capsules, granules, powders, solutions, suspensions, emulsions, syrups, lozenges, and the like. be able to. These various formulations are prepared according to a conventional method by adding an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a coloring agent, a flavoring agent, a solubilizing agent, and a suspending agent to the n-hexane extract of cheese as the main ingredient. It can be formulated by mixing adjuvants that are commonly used in the pharmaceutical formulation technical field, such as turbidity agents, surfactants, coating agents, and the like.

In addition, when the n-hexane extract of cheese is used as a pharmaceutical, for example, in the case of oral administration, the n-hexane extract of cheese can be ingested as it is. Tablets, granules, powders, capsules and powders can be used.

The content of the n-hexane extract of cheese in the drug can be appropriately determined according to the dosage form, usage, patient age, sex, degree of progression of cancer, degree of weight loss and other conditions.

In the embodiment of the present invention, as long as the inhibitory effect on vascular smooth muscle cell migration or the inhibitory effect on the progression of arteriosclerosis is not impaired, the drug containing n-hexane extract of cheese as an active ingredient may be used in combination with other drugs. do not have.

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.

(Preparation Example 1)
First, the commercially available "Meiji Hokkaido Tokachi (registered trademark) camembert cheese" manufactured by Meiji Co., Ltd. was immersed in liquid nitrogen and frozen, and the frozen camembert cheese was pulverized by a grinder into powder (hereinafter, this powder is called "Camembert cheese flour"). Next, 2 g of Camembert cheese powder was put into each of three 50 mL transparent centrifuge tubes. 20 mL of acetonitrile and 13 mL of n-hexane were then added to each tube, each tube was capped and shaken well to mix its contents. Subsequently, the three centrifuge tubes were set in a centrifuge and centrifuged at 2400 rpm for 10 minutes. Thereafter, the centrifuge tubes were removed from the centrifuge, and the lower acetonitrile phase was sucked from each centrifuge tube with a dropper and put into a 200 mL brown eggplant flask. Then, after removing the residue left in each centrifuge tube with a spatula, 10 mL of acetonitrile was added to each centrifuge tube, and the contents of each centrifuge tube were mixed well. Then, the three centrifuge tubes were again set in the centrifuge and centrifuged at 2400 rpm for 10 minutes. Thereafter, the centrifuge tubes were removed from the centrifuge, and the n-hexane phase of the upper phase was sucked from each centrifuge tube with a dropper and put into a 200 mL brown eggplant flask. Then, this eggplant flask was attached to a rotary evaporator to remove n-hexane from the eggplant flask to obtain the desired n-hexane extract of camembert cheese. Further, this n-hexane extract of Camembert cheese was dissolved in 20 mL of n-hexane, and the resulting solution was dispensed into five 5-mL brown glass vials of 1 mL each. Thereafter, while maintaining each glass vial at 40° C., nitrogen was flowed inside the glass vial to completely evaporate the n-hexane. Finally, after the inside of each glass vial was replaced with nitrogen, the lid of each glass vial was closed and they were stored frozen at -20°C.

In addition, general camembert cheese is manufactured as follows.

Prepare raw milk and pasteurize if necessary. Warm the raw milk. Lactic acid bacteria and rennet (milk-clotting enzyme) and, if necessary, mildew are added to the warmed raw milk. Whey (whey) is removed from the resulting curd to obtain cheese curds. Put the cheese curd into a mold and shape it. The molded cheese curd is salted. Here, if necessary, white mold may be sprayed on the surface of the cheese curd. The cheese curd is aged in a temperature- and humidity-controlled aging chamber. Examples of white mold used for producing white mold cheese include Penicillium camemberti, Penicillium candidum, Penicillium caseicolum, Geotrichum candidum, and the like. One type of white mold or two or more types may be used in the production of white mold cheese. The aging period is usually 1-4 weeks (eg 1-3 weeks). Moreover, you may heat after aging. Camembert cheese can thus be produced.

(Preparation Example 2)
After the second centrifugation in Preparation Example 1, the upper n-hexane phase is sucked from each centrifuge tube with a dropper, and the remaining lower acetonitrile phase is sucked up with a dropper, followed by the first centrifugation in Preparation Example 1. After separation, the lower acetonitrile phase was collectively added to a 200 mL brown eggplant flask. Then, this eggplant flask was attached to a rotary evaporator and acetonitrile was removed from the eggplant flask until the liquid amount became 50 g or less to obtain an acetonitrile extract of camembert cheese. Further, this acetonitrile extract of camembert cheese was divided into 5 equal parts, and each fraction was dispensed into 5 brown glass vials with a capacity of 15 mL. Each glass vial was then concentrated to dryness using a centrifugal evaporator. Finally, after the inside of each glass vial was replaced with nitrogen, the lid of each glass vial was closed and they were stored frozen at -20°C.

(Preparation Example 3)
Gorgonzola cheese in the same manner as in Preparation Example 1, except that "Meiji Hokkaido Tokachi (registered trademark) Camembert cheese" manufactured by Meiji Co., Ltd. was replaced with "Gorgonzola Dolce DOP" manufactured by Biraghi (Piedmont, Italy). -Hexane extract was obtained, and the n-hexane extract of the same Gorgonzola cheese was cryopreserved in the same manner as in Preparation Example 1.

Gorgonzola is generally produced as follows.

Prepare raw milk and pasteurize if necessary. Warm the raw milk. Lactic acid bacteria and rennet (milk-clotting enzyme) are added to the warmed raw milk. Whey (whey) is removed from the resulting curd to obtain cheese curds. Add blue mold to cheese curd and shape. After that, holes are drilled to allow the blue mold to grow evenly. Blue mold used for producing blue mold cheese includes, for example, Penicillin roqueforti. Blue mold used for producing blue mold cheese may be one type or two or more types. It may also be combined with mildew or yeast. Aging period is usually 1 week to 6 months (eg, 1 week to 3 months). Gorgonzola can thus be produced.

(Preparation Example 4)
After the second centrifugation in Preparation Example 3, the upper n-hexane phase is sucked from each centrifuge tube with a dropper, and the remaining lower acetonitrile phase is sucked up with a dropper, followed by the first centrifugation in Preparation Example 3. After separation, the lower acetonitrile phase was collectively added to a 200 mL brown eggplant flask. Then, this eggplant flask was attached to a rotary evaporator and acetonitrile was removed from the eggplant flask until the liquid volume became 50 g or less to obtain an acetonitrile extract of gorgonzola cheese. Further, this acetonitrile extract of Gorgonzola cheese was divided into 5 equal parts, and each fraction was dispensed into 5 brown glass vials having a capacity of 15 mL. Each glass vial was then concentrated to dryness using a centrifugal evaporator. Finally, after the inside of each glass vial was replaced with nitrogen, the lid of each glass vial was closed and they were stored frozen at -20°C.

(Preparation Example 5)
Phosphatidylethanolamine was detected from the n-hexane extract of Camembert cheese of Preparation Example 1 by liquid chromatography under the following conditions.

・Column: Accucore-RPMS 2.1×100 mm 2.6 μm
・Column temperature: 60°C
- Mobile phase A: 0.1% acetic acid solution of 10 mM ammonium acetate / acetonitrile = 90/10 (volume ratio) - Mobile phase B: 0.02% acetic acid solution of 2 mM ammonium acetate / 2-propanol / acetonitrile = 2/88/10 (capacity ratio)
・Flow rate: 0.25 mL/min ・Injection volume: 2 μL
- Gradient conditions: as described in Table 2 (development time: 43 minutes)
・Detection time: 16.4 to 20.6 minutes

By the way, the component analysis of the phosphatidylethanolamine obtained as described above was performed using a mass spectrometer (LC-QExactive (Orbtrap) manufactured by Thermo Fisher Scientific Co., Ltd.) under the following conditions. Phosphatidylethanolamines having chemical structures represented by chemical structural formulas (2) to (6) were detected. The component analysis was performed by converting the raw data (raw file) into an mzML file with 'MSConverter' and then fragment-analyzing it with 'Mass++'. In addition, a neutral loss spectrum of m/z: 141 was confirmed for each of the above phosphatidylethanolamines.

(a) Measurement mode: ESI-Positive mode (b) Ionization conditions Spray voltage: 3.0 kV
・Corona discharge current: -
・Vaporizer temperature: 400°C
・Capillary temperature: 250°C
・Nitrogen gas flow rate: 50arb
・S-sense level: 50
(c) Detection conditions Full scan resolution: 70,000
・AGC Target (MS1): 1×10 ⁶
・Measurement method: SIM-MS/MS mode
・AGC Target (MS2): 5×10 ⁴
・Stepped Collision: 15, 30, 45

(Preparation Example 6)
Phosphatidylethanolamine was detected from the n-hexane extract of Gorgonzola cheese of Preparation Example 3 by liquid chromatography under the following conditions.

・Column: Accucore-RPMS 2.1×100 mm 2.6 μm
・Column temperature: 60°C
- Mobile phase A: 0.1% acetic acid solution of 10 mM ammonium acetate / acetonitrile = 90/10 (volume ratio) - Mobile phase B: 0.02% acetic acid solution of 2 mM ammonium acetate / 2-propanol / acetonitrile = 2/88/10 (capacity ratio)
・Flow rate: 0.25 mL/min ・Injection volume: 2 μL
- Gradient conditions: as described in Table 3 (development time: 43 minutes)
・Detection time: 16.4 to 20.6 minutes

By the way, when the component analysis of the phosphatidylethanolamine obtained as described above was performed under the same conditions as those shown in Preparation Example 5, the compounds represented by the chemical structural formulas (4) to (6) above were found. A phosphatidylethanolamine with a chemical structure was detected.

(Preparation example 1)
The n-hexane extract of Camembert cheese prepared in Preparation Example 1 was dissolved in DMEM liquid medium (Dulbecco's Modified Eagle Medium, Nacalai Tesque Co., Ltd. DMEM High Glucose, 08458-45), 31.25 μg / mL, 62 DMEM liquid media containing 5 μg/mL, 125 μg/mL, 250 μg/mL and 500 μg/mL camembert cheese n-hexane extracts were prepared.

(Preparation example 2)
The acetonitrile extract of Camembert cheese prepared in Preparation Example 2 was dissolved in DMEM liquid medium (DMEM High Glucose, 08458-45 from Nacalai Tesque Co., Ltd.) to give 31.25 μg / mL, 62.5 μg / mL, 125 μg / DMEM liquid media containing Camembert cheese acetonitrile extracts of mL, 250 μg/mL and 500 μg/mL were prepared.

(Preparation example 3)
The n-hexane extract of Gorgonzola cheese prepared in Preparation Example 3 was dissolved in a DMEM liquid medium (DMEM High Glucose, 08458-45 from Nacalai Tesque Co., Ltd.) to give 31.25 μg/mL, 62.5 μg/mL, DMEM liquid media containing 125 μg/mL, 250 μg/mL and 500 μg/mL of Gorgonzola cheese n-hexane extract were prepared.

(Preparation example 4)
The acetonitrile extract of Gorgonzola cheese prepared in Preparation Example 4 was dissolved in DMEM liquid medium (DMEM High Glucose, 08458-45 from Nacalai Tesque Co., Ltd.) to give 31.25 μg/mL, 62.5 μg/mL, and 125 μg/mL. DMEM liquid media containing gorgonzola cheese acetonitrile extracts of mL, 250 μg/mL and 500 μg/mL were prepared.

(Preparation example 5)
The n-hexane extract of Camembert cheese prepared in Preparation Example 1 was dissolved in DMEM liquid medium (DMEM High Glucose, 08458-45 from Nacalai Tesque Co., Ltd.) to give 1 μg / mL, 10 μg / mL, 100 μg / mL A DMEM liquid medium containing camembert cheese n-hexane extract was prepared. The above three concentrations are certified as non-cytotoxic concentrations based on the results shown in Examples 1 and 2 and Comparative Examples 1 and 2 below.

(Preparation example 6)
The acetonitrile extract of camembert cheese prepared in Preparation Example 2 was dissolved in DMEM liquid medium (DMEM High Glucose, 08458-45 from Nacalai Tesque Co., Ltd.) to give 1 μg/mL, 10 μg/mL, and 100 μg/mL camembert cheese. A DMEM liquid medium containing acetonitrile extract was prepared.

(Preparation example 7)
The n-hexane extract of Gorgonzola cheese prepared in Preparation Example 3 was dissolved in DMEM liquid medium (DMEM High Glucose, 08458-45 from Nacalai Tesque Co., Ltd.), and 1 μg / mL, 10 μg / mL, 100 μg / mL A DMEM liquid medium containing gorgonzola cheese n-hexane extract was prepared.

(Preparation example 8)
The acetonitrile extract of Gorgonzola cheese prepared in Preparation Example 4 was dissolved in DMEM liquid medium (DMEM High Glucose, 08458-45 from Nacalai Tesque Co., Ltd.) to give 1 μg/mL, 10 μg/mL and 100 μg/mL of Gorgonzola cheese. A DMEM liquid medium containing acetonitrile extract was prepared.

(Example 1)
100 μL of DMEM-10% FBS (fetal bovine serum) liquid medium containing 5×10 ⁴ cells/mL of mouse aortic endothelial cells was seeded in a 96-well plate, and after 24 hours, cells from each well were DMEM-10% FBS liquid medium was removed. Next, 100 μL of the DMEM liquid medium containing Camembert cheese n-hexane extract of each concentration prepared in Preparation Example 1 was added to each well (3 wells were used for each concentration), and then the temperature was maintained at 37°C. Mouse aortic endothelial cells were cultured in an ambient _CO2 incubator for 20 hours. After 20 hours, 10 μL of 5 mg/mL MTT (Thiazolyl Blue Tetrazolium Bromide) PBS (phosphate buffered saline) solution was added to each well, and then again under a temperature environment of 37° C. for 20 hours. Mouse aortic endothelial cells were cultured in a _CO2 incubator. After 20 hours, the camembert cheese n-hexane extract-containing DMEM liquid medium was removed from each well. 100 μL of methyl sulfoxide (DMSO) was then added to each well. Finally, the absorbance at 490 nm of each well was measured using a plate reader to determine the cell viability of each well (here, the cell viability is DMEM containing no camembert cheese n-hexane extract (See Control Example 1 below). DMEM liquid medium containing 31.25 μg/mL camembert cheese n-hexane extract. The cell viability of the wells added with was 0.61, and the cell viability of the wells added with 62.5 μg/mL Camembert cheese n-hexane extract-containing DMEM liquid medium was 0.65, and 125 μg/mL. The cell viability of the wells to which the DMEM liquid medium containing the camembert cheese n-hexane extract of 250 μg/mL was added was 0.62, and the cell viability of the wells to which the DMEM liquid medium containing the camembert cheese n-hexane extract of 250 μg/mL was added. was 0.66, and the cell viability in wells to which 500 μg/mL camembert cheese n-hexane extract-containing DMEM liquid medium was added was 0.64 (see FIG. 1).

(Comparative example 1)
Examples except that the DMEM liquid medium containing camembert cheese n-hexane extract of each concentration prepared in Preparation Example 1 was replaced with the DMEM liquid medium containing camembert cheese acetonitrile extract of each concentration prepared in Preparation Example 2. After culturing mouse aortic endothelial cells in the same manner as in Example 1, the cell viability of each well was determined in the same manner as in Example 1.

As a result, the cell viability of the wells to which the DMEM liquid medium containing 31.25 μg/mL camembert cheese acetonitrile extract was added was 1.31, and the DMEM liquid medium containing 62.5 μg/mL camembert cheese acetonitrile extract was added. The cell viability of the wells containing 125 µg/mL camembert cheese acetonitrile extract was 1.27, and the cell viability of the wells containing DMEM liquid medium containing 125 µg/mL camembert cheese acetonitrile extract was 1.45, and 250 µg/mL camembert cheese acetonitrile extract. The cell viability of the wells to which the DMEM liquid medium was added was 1.46, and the cell viability of the wells to which the DMEM liquid medium containing 500 μg/mL camembert cheese acetonitrile extract was added was 1.54 (Fig. 1 reference).

(Example 2)
Except for replacing the DMEM liquid medium containing Camembert cheese n-hexane extract of each concentration prepared in Preparation Example 1 with the DMEM liquid medium containing Gorgonzola cheese n-hexane extract of each concentration prepared in Preparation Example 3, After culturing mouse aortic endothelial cells in the same manner as in Example 1, the cell viability of each well was determined in the same manner as in Example 1.

As a result, the cell viability of the wells to which the DMEM liquid medium containing 31.25 μg/mL of Gorgonzola cheese n-hexane extract was added was 0.56, and the DMEM containing 62.5 μg/mL of Gorgonzola cheese n-hexane extract was 0.56. The cell viability in the wells to which the liquid medium was added was 0.57, and the cell viability in the wells to which the DMEM liquid medium containing 125 μg/mL of Gorgonzola cheese n-hexane extract was added was 0.60 and 250 μg/mL. The cell viability of the wells to which the DMEM liquid medium containing the gorgonzola cheese n-hexane extract was added was 0.59, and the cell viability of the wells to which the DMEM liquid medium containing the gorgonzola cheese n-hexane extract of 500 μg/mL was added. was 0.64 (see Figure 1).

(Comparative example 2)
Example except that the DMEM liquid medium containing Camembert cheese n-hexane extract of each concentration prepared in Preparation Example 1 was replaced with the DMEM liquid medium containing gorgonzola cheese acetonitrile extract of each concentration prepared in Preparation Example 4. After culturing mouse aortic endothelial cells in the same manner as in Example 1, the cell viability of each well was determined in the same manner as in Example 1.

As a result, the cell viability of the wells to which the DMEM liquid medium containing 31.25 μg/mL gorgonzola cheese acetonitrile extract was added was 1.07, and the DMEM liquid medium containing 62.5 μg/mL gorgonzola cheese acetonitrile extract was added. The cell viability of the wells added with 125 μg/mL gorgonzola cheese acetonitrile extract-containing DMEM liquid medium was 1.23, and the cell viability of the wells added with 250 μg/mL gorgonzola cheese acetonitrile extract was 1.23. The cell viability of the wells to which the DMEM liquid medium was added was 1.28, and the cell viability of the wells to which the DMEM liquid medium containing 500 µg/mL of gorgonzola cheese acetonitrile extract was added was 1.55 (Fig. 1 reference).

(Example 3)
50 μL of DMEM liquid medium containing 5×10 ⁵ cells/mL of mouse aortic endothelial cells was seeded in each insert (upper chamber) of a 24-well insert and allowed to stand for 24 hours. Next, 50 μL of DMEM liquid medium containing Camembert cheese n-hexane extract at each concentration prepared in Preparation Example 5 was added to each insert, and 500 μL of 10 ng/mL PDGF (Platelet- Derived Growth Factor, platelet-derived growth factor)-containing DMEM-10% FBS liquid medium was added. After that, mouse aortic endothelial cells were cultured in a _CO2 incubator under a temperature environment of 37°C for 20 hours. After 20 hours, inserts and wells were washed twice with PBS. Mouse aortic endothelial cells on the back (underside) of the porous polycarbonate membrane of the insert were then treated with a solution of Trypsin in ethylenediaminetetraacetic acid (EDTA) for 30 minutes. Then, after the mouse aortic endothelial cells were well suspended in a trypsin-ethylenediaminetetraacetic acid solution, 100 μL of the cell suspension was added to an equal volume of DMEM-10% FBS liquid medium. Here, the DMEM-10% FBS liquid medium was placed in a 96-well plate in advance. Then, 20 μL of 5 mg/mL MTT (Thiazolyl Blue Tetrazolium Bromide) PBS (phosphate buffered saline) solution was added to each well of the 96-well plate, and again, the temperature was maintained at 37° C. for 4 hours under CO ₂ incubator for culturing mouse aortic endothelial cells. After 4 hours, the medium was removed from each well, 100 μL of dimethyl sulfoxide was added, and the absorbance at 490 nm was measured to determine the cell viability of each well. Mouse aortic endothelial cells were counted.

As a result, the cell viability of the wells to which the DMEM liquid medium containing 1 μg/mL camembert cheese n-hexane extract was added was 0.50, and the DMEM liquid medium containing 10 μg/mL camembert cheese n-hexane extract was added. The cell viability of the wells to which 100 μg/mL of Camembert cheese n-hexane extract was added was 0.41, and the cell viability of the wells to which the DMEM liquid medium containing 100 μg/mL camembert cheese n-hexane extract was 0.12 (see FIG. 3). That is, the camembert cheese n-hexane extract inhibited the migration of mouse aortic endothelial cells in a dose-dependent manner.

By the way, the procedure of the cell migration test is briefly summarized in FIG. In FIG. 2, the symbol CL indicates “mouse aortic endothelial cells”, the symbol CMl indicates “platelet-derived growth factor (PDGF)-containing DMEM-10% FBS liquid medium”, and the symbol CMp indicates “trypsin in EDTA solution”. , the code CMu indicates "cheese extract-containing DMEM liquid medium", the code IS indicates "insert", the code SM indicates "porous polycarbonate membrane", the code WL1 indicates "well", the code WL2 indicates " Other wells” are indicated.

(Comparative Example 3)
Same as Example 3, except that the DMEM liquid medium containing camembert cheese n-hexane extract of each concentration prepared in Preparation Example 5 was replaced with the DMEM liquid medium containing camembert cheese acetonitrile extract prepared in Preparation Example 6. After conducting a mouse aortic endothelial cell migration test, the cell viability of each well was determined in the same manner as in Example 3, and three locations in 96 wells were photographed, and the mouse aortic endothelial cells in each well were counted. bottom.

As a result, the cell viability of the wells to which the DMEM liquid medium containing 1 μg/mL camembert cheese acetonitrile extract was added was 0.98, and the cells in the wells to which the DMEM liquid medium containing 10 μg/mL camembert cheese acetonitrile extract was added The viability was 1.02, and the cell viability in wells added with DMEM liquid medium containing 100 μg/mL camembert cheese acetonitrile extract was 0.93 (see FIG. 4). That is, it could not be said that the camembert cheese acetonitrile extract inhibited the migration of mouse aortic endothelial cells.

(Example 4)
Example 3, except that the Camembert cheese n-hexane extract-containing DMEM liquid medium of each concentration prepared in Preparation Example 5 was replaced with the Gorgonzola cheese n-hexane extract-containing DMEM liquid medium prepared in Preparation Example 7. After conducting a mouse aortic endothelial cell migration test in the same manner as in Example 3, the cell viability of each well was determined in the same manner as in Example 3, and three locations in 96 wells were photographed, and the mouse aortic endothelial cells in each well were determined. counted.

As a result, the cell viability of the wells to which the DMEM liquid medium containing 1 μg/mL of gorgonzola cheese n-hexane extract was added was 0.98, and the DMEM liquid medium containing 10 μg/mL of gorgonzola cheese n-hexane extract was added. The cell viability of the wells to which 100 μg/mL of gorgonzola cheese n-hexane extract was added was 0.75, and the cell viability of the wells to which the DMEM liquid medium containing 100 μg/mL of gorgonzola cheese n-hexane extract was added was 0.49 (see FIG. 3). That is, the gorgonzola cheese n-hexane extract inhibited the migration of mouse aortic endothelial cells in a dose-dependent manner.

(Comparative Example 4)
Same as Example 3, except that the DMEM liquid medium containing Camembert cheese n-hexane extract of each concentration prepared in Preparation Example 5 was replaced with the DMEM liquid medium containing Gorgonzola cheese acetonitrile extract prepared in Preparation Example 8. After conducting a mouse aortic endothelial cell migration test, the cell viability of each well was determined in the same manner as in Example 3, and three locations in 96 wells were photographed, and the mouse aortic endothelial cells in each well were counted. bottom.

As a result, the cell viability of the wells to which the DMEM liquid medium containing 1 μg/mL of gorgonzola cheese acetonitrile extract was added was 0.66, and the cell viability of the wells to which the DMEM liquid medium containing 10 μg/mL of gorgonzola cheese acetonitrile extract was added was The viability was 1.23, and the cell viability in wells added with DMEM liquid medium containing 100 μg/mL of gorgonzola cheese acetonitrile extract was 0.80 (see FIG. 4). That is, it could not be said that the gorgonzola cheese acetonitrile extract inhibited the migration of mouse aortic endothelial cells.

(Control Example 1)
Mouse aorta was prepared in the same manner as in Example 3, except that the DMEM liquid medium containing Camembert cheese n-hexane extract of each concentration prepared in Preparation Example 5 was replaced with a DMEM-10% FBS liquid medium containing 10 ng/mL PDGF. After the endothelial cell migration test was performed, the cell viability of each well was determined in the same manner as in Example 3, and 96 wells were photographed at 3 locations, and mouse aortic endothelial cells in each well were counted. The viable cell rate in each of the above examples and comparative examples is obtained as a relative value with this count number as 1. That is, the cell viability of the wells according to this control example is 1 (see FIGS. 3 and 4).

(Control Example 2)
A mouse aortic endothelial cell migration test was performed in the same manner as in Example 3, except that the DMEM liquid medium containing each concentration of camembert cheese n-hexane extract prepared in Preparation Example 5 was replaced with dimethyl sulfoxide (DMSO). Then, the cell viability of each well was determined in the same manner as in Example 3, and 3 locations in 96 wells were photographed to count mouse aorta endothelial cells in each well. As a result, the cell viability in the well was 0.01 (see FIGS. 3 and 4).

(summary)
From the above results, it was confirmed that the camembert cheese n-hexane extract and the gorgonzola cheese n-hexane extract can dose-dependently suppress the migration of mouse aortic endothelial cells. Therefore, camembert cheese n-hexane extract and gorgonzola cheese n-hexane extract are expected to suppress the progression of arteriosclerosis in a dose-dependent manner.

The arteriosclerosis progression inhibitory composition, vascular smooth muscle cell migration inhibitory composition, and cheese n-hexane extract according to the present invention can inhibit the progression of arteriosclerosis and contribute to human health maintenance. Be expected.

CL mouse aortic endothelial cells CMl DMEM-10% FBS liquid medium containing platelet-derived growth factor (PDGF) CMp EDTA solution of trypsin CMu DMEM liquid medium containing cheese extract IS insert SM porous polycarbonate membrane WL1 well WL2 other wells

Claims (3)

A composition for inhibiting migration of vascular smooth muscle cells containing, as an active ingredient, an n-hexane extract of at least one mold type cheese selected from the group consisting of mildew type cheese and blue mold type cheese . 2. The composition for inhibiting vascular smooth muscle cell migration according to claim 1, wherein the n-hexane extract of mold type cheese dose-dependently inhibits vascular smooth muscle cell migration. 3. The composition for inhibiting migration of vascular smooth muscle cells according to claim 1 , wherein the mildew-type cheese is Camembert cheese and the blue-mold cheese is Gorgonzola cheese.

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