JP5013588B2 - Blood lipid improver - Google Patents

Description

  The present invention relates to a blood lipid improving agent containing cyclofructan, which is a cyclic inulooligosaccharide composed of fructose molecules cyclically linked by β-2,1 bonds, as an active ingredient.

In recent years, there are heart diseases and cerebrovascular disorders caused by abnormal lipid metabolism in the diseases that are listed as the top causes of death in Japan. These diseases are considered to be caused by so-called arteriosclerosis. Arteriosclerosis is thought to be caused by macrophages that are immune cells phagocytosing neutral fat, oxidized low-density lipoprotein (oxidized LDL), and the like, becoming foam cells and collecting in the blood vessels. In addition, arteriosclerosis is caused by hyperlipidemia, a condition in which neutral fat and cholesterol accumulate excessively. In other words, a state in which neutral fat, cholesterol, and the like in the blood increase beyond the normal range, so-called hyperlipidemia, is the first trigger that causes diseases such as heart disease and cerebrovascular disorder. However, since hyperlipidemia is a disease that takes a long time to cure and requires continuous treatment, it is highly safe and does not cause side effects even when taken on a daily basis. There is a need for a material that has no problem even if it is added to, and has an effect of reducing neutral fat and the like.
On the other hand, only a small amount of knowledge has been obtained about the bioregulatory function of cyclofructan, which is a cyclic inulooligosaccharide composed of a fructose molecule linked in a cyclic manner with β-2,1 bonds. These include improving the intestinal function by increasing short-chain fatty acids in the intestine (see Patent Document 1 below), and stimulating mucosal immunity to induce secretory IgA production (see Patent Document 2 below) ), And a dry skin recovery effect based on a decrease in stratum corneum moisture (see Patent Document 3 below). In addition, an effect of improving drug permeability in mucosal tissues (see Patent Document 4 below) is also disclosed.
However, it has never been known that cyclofructan has a blood lipid improving action.
JP 2005-154388 A JP 2005-179195 A JP-A-7-53347 JP 2005-523299 A

  Accordingly, an object of the present invention is to provide a new blood lipid improving agent.

The present inventors have studied in view of the above points. As a result, it was found that cyclic inulooligosaccharides (cyclofructans) composed of fructose molecules cyclically linked by β-2,1 bonds have an excellent blood lipid improving action. Furthermore, it has been found that it has an effect of adjusting the value of neutral fat among blood lipids to a normal value or a range closer to a normal value.
The fact that cyclofructan functions as a blood lipid improving agent has not been reported so far, and is a completely new finding that cannot be recalled from the prior art.

That is, the blood lipid improving agent of the present invention comprises cyclofructan as an active ingredient as described in claim 1.
The blood lipid improving agent according to claim 2 has the function of improving neutral fat, in particular, in the blood lipid improving agent according to claim 1.
The blood lipid improving agent according to claim 3 is the blood lipid improving agent according to claim 1 or 2, wherein the cyclofructan is at least one selected from cyclic inulohexaose and cyclic inuloheptaose. It is.

  The present invention is a new blood lipid improving agent comprising cyclofructan as an active ingredient, and has a high versatility that can be widely applied to foods and drinks and pharmaceuticals. Ingestion of the blood lipid improving agent in the present invention can improve blood lipids, particularly neutral fat, and can lead to prevention or improvement of various diseases related to hyperlipidemia.

The blood lipid improving agent comprising the cyclofructan of the present invention (hereinafter abbreviated as “CF”) as an active ingredient effectively suppresses an increase in blood lipid that causes arteriosclerosis or causes excess blood in the blood. It has a function of improving lipids in blood, such as reducing lipids or increasing or maintaining lipids that are preventive factors for arteriosclerosis. Generally, blood lipids include cholesterol, neutral fat, phospholipid, free fatty acid, and the like, but the blood lipid improving agent of the present invention works particularly effectively on neutral fat.
In the present invention, CF functioning as an active ingredient of a blood lipid improving agent means a cyclic inulooligosaccharide composed of a fructose molecule linked in a cyclic manner with β-2,1 bonds, and consists of 6 fructose molecules. Cyclic inulohexaose (hereinafter abbreviated as “CF6”), cyclic inuloheptaose composed of 7 fructose molecules (hereinafter abbreviated as “CF7”), cyclic inulooctaose composed of 8 fructose molecules ( (Hereinafter abbreviated as “CF8”). These are all white powders, have extremely high solubility in water, and are slightly soluble in water-containing organic solvents. CF, for example, is a cyclic inuro-oligosaccharide-forming enzyme to inulin, which is the main component of carbohydrates obtained from the roots and rhizomes of asteraceae, liliaceae, iridaceae, orchidaceae such as Jerusalem artichoke, chicory, burdock, and dahlia. It can be obtained by the action of a certain cycloinulooligosaccharide fractanotransferase (hereinafter abbreviated as “CFTase”).

  Hereinafter, a method for obtaining and purifying CF will be described in detail. CTFase necessary for obtaining CF is CFC1 (KIM HWA-Young and YONG-JIN CHOI, J. Microbiol. Biotechnol, vl. 8, no. 3, p. 251-257, 1998) belonging to Bacillus macerans. ), OKUMZ 31B belonging to Bacillus circulans (Mishio Kawamura and Takao Uchiyama, Carbohydr Res, vl. 260, p.297-304, 1994), MCI-2554 belonging to Bacillus circulans (Sachiko Kushibe and Kaori Mitsui) , Biosci. Biotech. Biochem., Vl. 59, no. 1, p31-34, 1995, JP 7-41500 A), MG-CF6 belonging to Paenibacillus polymyxa (FERM P-19158, JP It can be produced using a CFTase producing microorganism such as 2004-242623. CF can be obtained from the culture supernatant after culturing such a CPTase-producing microorganism in a medium containing inulin, sterilizing it using centrifugation or the like. CF can also be obtained by separately treating crude CFTase prepared from a culture supernatant of a CFTase-producing microorganism or purified CFTase with inulin. CF obtained by such a method is a mixture composed of CF6, CF7, CF8, etc., and these are activated carbon column chromatography, ion exchange chromatography, gel filtration (Mishio Kawamura, Ikuo Uchiyama, Starch Science, Vol.39, p.109-116,1992, KIM HWA-Young et al., J. Microbiol. Biotechnol, vol.6, no.6, p.397-401, 1996) etc. Can do.

  As the blood lipid improving agent of the present invention, the CF mixture obtained by the above method may be used as an active ingredient as it is, or a single separated and purified CF compound may be used. The above-mentioned single CF compound may be mixed and used at a desired mixing ratio, and the blood lipid improving action of the present invention can be exerted regardless of which method is used.

Examples of foods and drinks to which the blood lipid improving agent in the present invention is added include, for example, instant foods (instant noodles, cup noodles, retort / cooked foods, canned foods, microwave foods, instant miso soup / soup, canned soups, freezes, etc. Dry foods), carbonated drinks, fruit juice drinks such as citrus fruits (grapefruit, orange, lemon, etc.), fruit drinks, fruit drinks with fruits, tomatoes, peppers, celery, cucumbers, carrots, potatoes, asparagus and other vegetables Vegetable drinks, soy milk and soy milk drinks, coffee drinks, tea drinks (green tea drinks, oolong tea drinks, tea drinks, etc.), sports drinks, nutrition drinks, taste drinks such as tobacco, sweets (caramel candy, chewing gum, Chocolate, cookie biscuits, cake pie, snack crackers, Japanese confectionery -Rice confectionery / bean confectionery, dessert confectionery, etc.) Basic seasonings (soy sauce, miso, sauces, tomato processed seasonings, mirins, vinegars, sweeteners, etc.), complex seasonings / foods (flavor seasonings, Cooking mixes, curry ingredients, sauces, dressings, noodle soups, spices, etc., milk and dairy products (butter, margarines, mayonnaise, vegetable oils, milk and processed milk, milk drinks, yogurt , Lactic acid bacteria beverages, cheese, ice cream, prepared milk powder, cream, etc.), frozen food (material frozen food, semi-cooked frozen food, cooked frozen food, etc.), seafood canned and pasted, fruit canned and pasted, Processed marine products (fish ham / sausage, marine products, marine delicacies, marine dried products, boiled fish, etc.), livestock canning / pastes, canned meat, jam / marmalade , Pickles, boiled beans such, agricultural dry goods, serial (grain processed products) processed agricultural products such as, baby food, such as commercially available foods such as sprinkled-Ochazuke glue and the like. In the present invention, a concentrated beverage obtained by concentrating a liquid beverage or a powdered beverage obtained by pulverizing a liquid beverage or a concentrated beverage by spray drying or spray drying can be included in the beverage. Here, the powdered beverage refers to a food or drink provided in the form of a powder at the time of sale and dissolved in water or the like to an appropriate concentration at the time of drinking.
The blood lipid improving agent of the present invention can be used to prevent and ameliorate various diseases particularly related to hyperlipidemia, and, as necessary, beauty food, food for the sick, or food for specified health use It is preferable to apply to functional foods and beverages.

  In addition, when used as a medicine, it is not particularly limited as long as it can be contained in the mouth with a medicine stored in the Japanese Pharmacopoeia. Examples of the preparation form include oral solids such as tablets and granules. Examples include preparations and oral liquid preparations such as oral liquids and syrups.

  The method for blending the blood lipid improving agent of the present invention into foods and drinks and pharmaceuticals is not particularly limited. For example, when the compounded object is in a liquid form or semi-solid form, the blood lipid improving agent of the present invention is added as it is or in the form of an aqueous solution dissolved in water at the preparation stage, and homogenized. To do so. It is also possible to add the blood lipid improving agent of the present invention as a dispersion liquid dispersed in a water-containing organic solvent such as alcohol water or an organic solvent such as ethanol, and disperse it with sufficient stirring. In addition, it is good also as a powder form by drying the preparation obtained by doing in this way using a spray dryer, a freeze dryer, etc. Further, when the composition is in a solid form, the blood lipid improving agent of the present invention is added as it is or in the form of an aqueous solution dissolved in water at the preparation stage, and the mixture is homogenized. Just do it. In addition, when the blood lipid improving agent of the present invention is blended with a poorly water-soluble blending object, a water-containing organic solvent such as alcohol water or an organic solvent such as ethanol is added to the food or drink as necessary to disperse it. Alternatively, the blood lipid improving agent of the present invention may be added thereto, and the blood lipid improving agent may be sufficiently stirred to disperse or dissolve the blood lipid improving agent.

  Although the compounding quantity of the blood lipid improving agent of this invention with respect to food-drinks, a pharmaceutical, etc. is not restrict | limited in particular, It is preferable to set a compounding quantity suitably with target food-drinks. Generally, it is preferably 0.1 to 100% by weight in the final product, more preferably 0.5 to 95% by weight, and still more preferably 5 to 90% by weight.

The blood lipid improving agent of the present invention may be ingested by being mixed with food or drink as described above, but can also be ingested as it is. The intake amount varies depending on the intake form, age, weight, etc., but is not particularly limited.
The dosage form in the case where the blood lipid improving agent of the present invention is formulated and ingested alone may be any form as long as it contains CF as an active ingredient. It may be liquid.
The blood lipid improving agent of the present invention is represented by other blood lipid improving agents such as polyphenols such as catechins, medium chain fatty acids, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and the like. No problems arise when used in combination with unsaturated fatty acids. When used in combination with other blood lipid improving agents, a better blood lipid improving action can be expected.
In addition, when formulating the blood lipid improving agent of the present invention in foods and drinks or when formulating alone, as necessary, extenders, antioxidants, colorants, flavors, flavoring agents, surfactants, You may use it combining suitably various well-known various additives, such as a solubilizing agent, a preservative, a sweetener, saccharides other than CF, a sour agent, and vitamins.

  The blood lipid improving agent in the present invention can exert the effects of the present invention by ingestion in a food or drink as described above or by ingestion alone.

  The blood lipid improving agent containing CF of the present invention as an active ingredient is useful for preventing various diseases caused by an increase in blood lipid. In particular, it is possible to improve blood lipids and prevent hyperlipidemia without dietary restrictions or medication.

  Hereinafter, the present invention will be described in more detail with reference to production examples, test examples and formulation examples. However, the present invention is not limited to this.

Production Example 1: Production method of CF mixture Inulin 4%, yeast extract 0.2%, sodium nitrate 0.5%, magnesium sulfate 0.05%, potassium chloride 0.05%, monopotassium phosphate 0.05% 200 mL of a medium containing ferric chloride 0.001% was adjusted to pH 7.5 and steam sterilized at 120 ° C. for 15 minutes. One platinum loop of Paenibacillus polymixer MG-CF6 (FERM P-19158) was inoculated into this sterilized medium, and cultured with shaking at 200 rpm and 35 ° C. for 48 hours. After completion of the culture, the cells were removed by centrifugation to obtain a culture supernatant. 4000 mL of the culture supernatant obtained by repeating the above operation was concentrated under reduced pressure to obtain crude CF-containing liquid sugar (about 500 mL, 72 g as a solid content).
After adjusting the pH of this crude CF-containing sugar solution to 4.5 with 1 M citric acid, 0.5 mL (about 1000 units) of exo-type and endo-type inulinase mixed enzyme (trade name Fructozyme L, manufactured by Novo Nordisk) was added, It was allowed to act for 1 hour at 60 ° C. This was heated at 100 ° C. for 10 minutes to deactivate the enzyme, and the denatured protein was removed by centrifugation. When the obtained reaction solution (CF-containing enzyme treatment liquid) was analyzed by high performance liquid chromatography, only CF, fructose, and glucose were found as sugars.
This CF-containing enzyme treatment solution was directly applied to an activated carbon column (40 × 900 mm) equilibrated with water. The column was washed with 6000 mL of water and then eluted with the same amount of 30% (v / v) aqueous ethanol. The ethanol aqueous fraction was concentrated under reduced pressure and lyophilized to obtain about 30 g of a CF mixture (composition ratio: CF6: 61.7 wt%, CF7: 30.7 wt%, others: 7.6 wt%).

Production Example 2: CF Purified Product 30 g of the CF mixture obtained in Production Example 1 was dissolved in a small amount of a 70% aqueous ethanol solution. This solution was applied to a QAE-Toyopearl column (40 × 900 mm) equilibrated with an aqueous ethanol solution having the same concentration to separate CF6 and CF7. These were freeze-dried to obtain pure powders of 15 g CF6 and 7 g CF7.

Test Example 1: Blood lipid improving action test by CF The blood lipid improving action by CF was examined according to the following test method.
(Test method)
Four-week-old SD strain (Sprague-Dawley) male rats were bred on a solid diet (Lab Diet 5L37, Japan SLC) for 3 days, then divided into 2 groups of 8 mice so that their body weights were equal, and the control (C) Group and cyclofractane added (CF) group. The C group was reared with a high fat diet supplemented with 1 wt% cholesterol and 15 wt% lard, and the CF group with 5 wt% cyclofructan added to the high fat diet. Each group was allowed free intake of each feed (Table 1) and water for 14 days. After the breeding, whole blood was collected after overnight fasting and blood neutral fat was measured. The breeding room temperature was 23 ° C., with a light period of 8:00 to 20:00 and a dark period of 20:00 to 8:00.
Table 1 shows the feed preparation ratios used in the examples.

(Test results)
As a result of measuring the blood triglycerides in each of the C group and the CF group, as shown in FIG. 1, the blood triglyceride in the CF group is significantly lower than that in the C group. It was found that the neutral neutral fat value was 149.8 mg / dL, whereas the CF group was markedly suppressed to 87.1 mg / dL.
The same effects as above were confirmed for each of CF6 and CF7 obtained in Production Example 2.

  Below, the formulation example which mix | blended the blood lipid improving agent in this invention with food-drinks etc. is shown. The food / beverage products containing the blood lipid improving agent of the present invention can be ingested deliciously without impairing the original taste / fragrance of the food / beverage products.

Formulation Example 1: Green tea beverage containing blood lipid improving agent 900 mL of water was heated to 60 ° C., and 30 g of green tea leaves were added thereto and extracted for 6 minutes. After removing the tea husk with a 30-mesh strainer and cooling to 30 ° C. or lower, it is clarified by filter paper filtration (industrial filter paper No. 26: ADVANTEC, collected particle size = 3 μm), and 760 mL of extract liquid Got. This green tea extract is diluted with ion-exchanged water to a drinking concentration (catechin concentration 60 mg%), L-ascorbic acid is added to 0.03% by weight, and then pH 6.1 to 6. It adjusted to the range of 3 and was set as the preparation liquid. 5% by weight of the CF mixture obtained in Production Example 1 was added to this prepared solution and stirred well. These were hot-packed under a temperature condition of 80 ° C. or higher, and then sterilized by retort sterilization at 121 ° C. for 10 minutes (F ₀ = 10 or higher). After cooling these to room temperature, a green tea beverage containing CF was obtained.

Formulation Example 2: Oral solution containing blood lipid improving agent
CF7 10g obtained in Production Example 2
Carrot dry extract 214mg
Dried licorice extract 50mg
Ursodeoxycholic acid 25mg
D-sorbitol 50g
25g white sugar
Polyoxyethylene hydrogenated castor oil 3g
Propylene glycol 5mL
Sodium benzoate 600mg
Butyl paraoxybenzoate 10mg
Citrate buffer appropriate amount Ion-exchanged water was added to the above components to make the total amount 300 mL, and an internal solution containing CF7, which was a purified CF product obtained in Production Example 2, was prepared.

  As described above, the cyclofructan (CF) of the present invention can be added to various foods and drinks.

It is a figure which shows the amount of blood triglycerides of C group and CF group in an Example.

Claims (2)

An additive for improving blood neutral fat, comprising cyclofructan as an active ingredient. The additive for improving blood neutral fat according to claim 1 or 2, wherein the cyclofructan is at least one selected from cyclic inulohexaose and cyclic inuloheptaose.

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