JP5577019B2 - Orally administered composition - Google Patents

Description

  The present invention relates to a composition comprising a carotenoid and a sphingolipid that is effective for reducing body fat, reducing neutral fat, and the like and can be taken as food, pharmaceuticals, and feed.

  From April 2008, medical insurers (National Health Insurance and Employee Insurance) will be obliged to conduct medical examinations and health guidance projects focusing on metabolic syndrome for insured persons and dependents over 40 years of age. After five years, the outcome is judged and a financial penalty is imposed on health insurers with poor results. The Ministry of Health, Labor and Welfare expects that one of two middle-aged men has metabolic syndrome, and about 20 million people fall into the metabolic syndrome and reserve group.

  In recent years, research on the effects of fat on the body has progressed, and it has been suggested that visceral fat accumulation is not only obese, but also highly correlated with lifestyle-related diseases, hyperlipidemia, impaired glucose tolerance, hypertension, etc. It is believed that not accumulating fat can prevent many diseases. The pathological condition based on these ideas is metabolic syndrome. To reduce visceral fat, it is necessary to change the living environment such as improving meals and performing daily exercise. However, due to westernization of meals and changes in lifestyle, meals become high in calories and it is difficult to secure time for exercise.

  At present, in Japan, attention has been focused on preventive medicine in order to reduce the cost of medical care that will increase as the population ages. When the above diseases occur, a large amount of treatment costs and treatment time are required. Therefore, various health foods and supplements are on the market to reduce visceral fat from the viewpoint of preventive medicine to reduce the risk of developing the disease. .

  However, it is difficult for these health foods to be expected to have a sufficient effect when taken in a normal manner, and there is a drawback that side effects are increased although a certain level of effect can be expected when taken in large amounts. As described above, there is a strong demand for the development of a food having an action of reducing visceral fat effectively and safely with a small amount of intake.

Catechin and astaxanthin are examples of existing foods that have the effect of reducing such body fat, particularly visceral fat. Catechin is contained in green tea, black tea, chocolate and the like, and is said to be effective in burning body fat (Patent Document 1). However, increasing the content increases astringency, so the addition to the product is greatly restricted. Astaxanthin also exhibits a body fat burning effect (Patent Document 2), but astaxanthin itself is difficult to be taken into the body, and since the effect alone is not high, it is difficult to take an effective amount for fat reduction. .
JP 2002-326932 A JP 2007-153845 A

  In view of the current situation as described above, provision of foods, pharmaceuticals, and feeds that have an effect of reducing body fat such as visceral fat and subcutaneous fat with a normal intake amount has been awaited. An object of the present invention is to provide foods, pharmaceuticals, and feeds that are highly safe and have a body fat reducing action and a neutral fat reducing action.

  As a result of intensive investigations to solve such problems, the present inventors, when carotenoids and sphingolipids are used in combination, are capable of exerting effects beyond the functionality expected when used alone due to their synergistic action. The present invention has been completed. It was also found that these combinations can be easily extracted and concentrated when citrus plants are used as raw materials.

  That is, the first of the present invention is an orally administered composition comprising a carotenoid and a sphingolipid and having a body fat and / or neutral fat reducing action, preferably a composition in which the carotenoid is cryptoxanthin. The carotenoid and / or sphingolipid is derived from citrus, and preferably the citrus is Wenzhou mandarin.

  The second aspect of the present invention is characterized in that a citrus plant is squeezed and a composition containing carotenoid and / or sphingolipid is obtained from the residue, or an enzyme is added to the citrus plant and the carotenoid is treated with the enzyme. And / or obtaining a composition containing a sphingolipid, or adding an organic solvent to a citrus plant and extracting the composition containing a carotenoid and / or a sphingolipid in the organic solvent. The gist of the method for producing any one of the compositions described above.

  The third aspect of the present invention is the gist of foods, pharmaceuticals and feeds comprising the aforementioned oral administration composition.

  According to the present invention, a high body fat reducing action / neutral fat reducing action is exhibited, the safety is high, and it can be added to various foods, pharmaceuticals, and feeds.

  The present invention is described in detail below.

  The carotenoid in the present invention is not particularly limited. Of fatty acid esters. Examples of fatty acid esters include the above-mentioned carotenoids such as palmitoyl ester, myristoyl ester, and lauryl ester. Among these, cryptoxanthin and fatty acid esters thereof are desirable because of their high body fat reducing action and neutral fat reducing action.

  The cryptoxanthin in this invention is not specifically limited, For example, (alpha) -cryptoxanthin, (beta) -cryptoxanthin, and these fatty acid ester bodies and structurally acceptable derivatives are also contained. The fatty acid length of the fatty acid ester is not particularly limited, and examples thereof include fatty acid esters such as lauric acid (C12), myristic acid (C14), palmitic acid (C16), and stearic acid (C18). Moreover, as a supply source of these cryptoxanthins, it is preferable that it is citrus, and it is especially preferable that it is derived from Unshu mandarin orange which is a large amount of production and is an ancient Japanese fruit.

  The sphingolipid in the present invention is a group of compounds in which an amino alcohol having about 14 to 24 carbon atoms called sphingosine or a long-chain amino alcohol having a similar structure to this and a fatty acid such as a long-chain fatty acid are amide-bonded. In other words, compounds in which saccharides are bound to them, and compounds in which phosphate groups are bound are also included. The fatty acid may be either a saturated fatty acid or an unsaturated fatty acid, may be linear or branched, and one or more hydroxyl groups may be substituted. In addition, sugar, phosphoric acid, sialic acid, choline, ethanolamine and the like may be bonded to the hydroxyl group of amino alcohol. As the saccharide, it is preferable that glucose, mannose, galactose, lactose and the like are bonded, and it is particularly preferable that glucose is bonded. Those having a phosphate group bonded thereto include inositol via a phosphate group, and also sphingolipids having a sugar bonded thereto. The present invention is not intended to have a specific chain length or structure, but includes all those called sphingolipids. Moreover, the origin may be a conventionally known sphingolipid or a derivative thereof derived from a natural product or obtained by chemical synthesis. When these sphingolipids are used for foods, they can be obtained from various mushrooms such as rice, wheat, milk, corn, beet, konjac, apple, tamogitake and maitake, and acetic acid bacteria. Although it can be used as it is, a sphingolipid derived from citrus is preferred, and a sphingolipid derived from Wenzhou mandarin orange is particularly preferred. Citrus-derived sphingolipids contain a large amount of free-type sphingolipids that do not contain sugars, and therefore, it is preferable to use those derived from them because of their strong effects on reducing body fat and neutral fat.

  Citrus in the present invention includes plants belonging to the citrus family and the like. More specifically, Iyokan, Wenzhou mandarin orange, Orange, Kabos, Kawabata, Kishumikan, Kiyomi, Kumquat, Grapefruit, Gecki, Sanpokan, Shikuwasa, Jabara, Sweety, Sudachi, Daidai, Tachibana, Dekopon, Natsudaidai, Hassaku, Valencia Orange Illustrative examples include evening birch, hyuganatsu, buntan, ponkan, mandarin orange, yatsushiro, yuzu, lime, lemon, karatatachi and the like.

  In the present invention, the citrus plant used as a raw material for extraction and enzyme treatment is not limited to the part of the citrus plant, but among them, it is preferable to use the fruit as a part having a high content. The fruits of citrus plants may be used as they are, or those obtained by subjecting them to physical treatments such as grinding, crushing, crushing, heating, dehydration, and drying. These treatments are preferable because enzyme treatment efficiency and extraction efficiency are increased. Furthermore, in the present invention, it is preferable to use a fruit juice residue. The fruit juice residue is obtained by squeezing the fruit with an inline squeezer, chopper helper squeeze machine, brown squeeze machine, etc., then filtered or sieved with a paddle type or screw type finisher, etc. It is prepared by collecting the juice residue prepared. Moreover, the squeezed residue of the citrus plant used by this invention can also use the squeezed residue which is a fine pulp content obtained by centrifuging the fruit juice after squeezing with a squeezing machine. Such a fine pulp content is a preferable raw material because of its higher content of cryptoxanthin and sphingolipid.

  Examples of the organic solvent used in the extraction step in the present invention include alcohols such as methanol, ethanol, propanol and isopropanol, ketones such as acetone and methyl ethyl ketone, esters such as methyl acetate and ethyl acetate, and halogenated carbonization such as acetonitrile and chloroform. Examples include hydrogens, aliphatic hydrocarbons such as pentane and hexane, aromatic hydrocarbons such as benzene and toluene, ethers, pyridines, polyethers, and supercritical carbon dioxide. Among these, in the present invention, since it is used for food applications, it is particularly preferable to use acetone, ethanol, hexane, or supercritical carbon dioxide from the viewpoint of safety. These organic solvents may be used alone or as a mixture. In addition, additives such as water and a surfactant can be added to the organic solvent.

  The amount of the organic solvent used for extraction is not particularly limited as long as it is sufficient to extract cryptoxanthin and sphingolipid from the extract. Specifically, it is 0.5 to 100 times, preferably 1 to 50 times the solid content weight of the extract. Although the extraction temperature depends on the boiling point of the solvent used, for example, when ethanol is used, it is preferably 0 ° C. to 80 ° C. If the extraction temperature is lower than this range, the extraction efficiency is lowered. If the extraction temperature is higher than this range, the solvent is volatilized and only the amount of energy used is increased. The extraction time is not particularly limited, but preferably 10 minutes to 24 hours from the viewpoint of extraction efficiency and workability.

  The extraction operation is not limited to a single batch operation. A new solvent can be added again to the residue after extraction to perform an extraction operation, or the extraction solvent can be brought into contact with the extract multiple times. That is, as the extraction operation, any of batch operation, semi-continuous operation, and countercurrent other-stage contact operation can be used. Moreover, you may use well-known extraction methods, such as Soxhlet extraction and reflux extraction. Separation and removal of the residue after extraction may be performed by a known method. Specifically, suction filtration, filter press, cylinder press, decanter, centrifuge, filtration centrifuge, or the like may be used. Further, impurities may be subsequently removed. Examples of the impurity removal method include water washing, organic solvent washing, silica gel column, resin column, reverse phase column and the like, activated carbon treatment, partitioning with solvents of different polarity, recrystallization method, vacuum distillation method and the like. .

  The extract thus obtained is then subjected to treatments such as alkali treatment, concentration and purification with a column, drying, pulverization, mixing with a carrier, emulsifier, etc., to all foods, pharmaceuticals and feeds. Can be used.

  In the present invention, the enzyme used for producing an enzyme-treated product of citrus plants is not particularly limited as long as it can decompose organic substances contained in citrus, particularly biopolymers constituting cell walls. Amylase, glucoamylase, cellulase, hemicellulase, pectinase, mannanase, xylanase, protease, peptidase, lipase, maceration enzyme (cell wall degrading enzyme) and the like are used. Among these, carbohydrate hydrolase cellulase, hemicellulase, pectinase, mannanase, xylanase, and malation enzyme are preferable enzymes because of their high efficiency of concentrating sphingolipids. As the enzyme agent to be added, these purified enzymes may be used, or microbial cells or cultures exhibiting these activities, or crudely purified products thereof may be used. Commercially available enzymes can also be used. For example, for pectinase, Sumiteam PX (manufactured by Shinnippon Chemical Co., Ltd.), Sumiteam SPC (manufactured by Shinnippon Chemical Co., Ltd.), pectinex SRL (manufactured by Novozymes Japan Ltd.) ), Sumiteam PMAC (manufactured by Shinnippon Kagaku Kogyo Co., Ltd.), etc., and cellulosin GM5 (manufactured by HIBI), cellulosin HC (manufactured by HIBI), etc. can be used as the hemicellulase. Cellulase Y-NC (manufactured by Yakult Pharmaceutical Co., Ltd.), Entilon MCH (Pinto Kasei Kogyo Co., Ltd.), Cellulase R10 (manufactured by Yakult Pharmaceutical Co., Ltd.), Sumiteam CAP (manufactured by Shin Nippon Chemical Industry Co., Ltd.), Cellulase TP-3 (Kyowa Kasei Co., Ltd.) can be used, As the protease, protease M (manufactured by Amano Enzyme Co., Ltd.), orientase 20A (manufactured by HBI Co., Ltd.) and the like can be used.

  These enzymes may be used alone, or two or more kinds of enzymes may be mixed and used. The amount of the enzyme to be added is not particularly limited, and may be added according to the reactivity of the enzyme. For example, if pectinase is used, it is preferably 1 to 100,000 units, more preferably 10 to 10,000 units, with respect to 100 g of juice residue.

  After the enzyme is added, the enzyme and the extract are uniformly mixed by stirring or the like to allow the enzyme reaction to proceed. As the reaction temperature at this time, it is desirable to carry out the reaction under conditions where the enzyme is not inactivated and is not easily spoiled, or where the sphingolipid is not lost. Specifically, the temperature is 0 ° C to 90 ° C, preferably 0 ° C to 80 ° C, and more preferably 0 ° C to 70 ° C. Needless to say, the pH of the reaction is preferably carried out under the optimum conditions of the enzyme, and it is preferably pH 2 to 12, preferably pH 2.5 to 8. The reaction time depends on the squeezed residue used and the amount of the enzyme, but it is usually preferable to set the reaction time to 1 to 48 hours. In the reaction, the reaction may be performed while stirring the reaction product, or may be a stationary reaction.

  After completion of the enzyme treatment, the enzyme-treated reaction product may be used as it is, or a product that has undergone some processing may be used. Specifically, a residue obtained by solid-liquid separation of the reaction product, a product obtained by drying the residue obtained by solid-liquid separation, or a product obtained by drying the reaction product as it is without solid-liquid separation may be used. In addition, a water washing method that can easily remove reaction products such as sugars produced by enzyme treatment by solid-liquid separation of the enzyme-treated reaction product, addition of water, and solid-liquid separation again. When used, it is preferable because impurities can be easily removed. Further, impurities may be subsequently removed. Examples of the impurity removal method include chemical treatment such as water washing, organic solvent washing, and alkali treatment.

  The composition of the present invention obtained as described above exhibits various functionalities, and when taken orally, it is represented by body fat such as visceral fat and subcutaneous fat, and triglyceride among them. It has a reducing effect on neutral fat and the like.

  As a method of ingesting the composition of the present invention, the composition may be ingested alone, or powder, tablet, granule, capsule, soft capsule, gel, paste, syrup, suspension, emulsion It may be processed into a drink and taken. Ingestion guidelines should be selected in a timely manner according to the subject's age, gender, weight, etc. Usually, for adults, 0.01 mg to 1 g of carotenoid and 0.01 mg of sphingolipid per day 10 to 10 g, preferably 0.1 mg to 100 mg of carotenoid and 0.1 mg to 1 g of sphingolipid may be taken.

  Another invention of the present invention is a food and drink and a medicine containing the composition of the present invention described above. As pharmaceuticals, injections, infusions, powders, tablets, granules, capsules, pills, intestinal solvents, suspensions, syrups, oral liquids, lozenges, emulsions, liquids for external use, poultices, nasal drops, It can be taken in the form of ear drops, eye drops, inhalants, ointments, lotions, suppositories, enteral nutrients and the like. These can be used alone or in combination of two or more depending on the symptoms. The content of the composition of the present invention may be appropriately determined in the range of 0.1% to 90%, but it is preferable to design the formulation so that the above-mentioned guideline for daily intake can be ingested.

  The food and drink of the present invention includes all foods and / or beverages such as foods for specified health use, health foods, functional foods and quasi drugs in addition to general foods. The food and / or beverage is not particularly limited. For example, in addition to the above-mentioned pharmaceutical form, bread, udon, buckwheat, rice, and other staple foods, cheese, wiener, sausage, ham, seafood Processed foods, ice cream, cookies, cakes, jelly, pudding, candy, chewing gum, yogurt, gummy, chocolate, biscuits and other sweets, soft drinks, seasonings, alcoholic beverages, energy drinks, coffee, tea And beverages such as milk, fruit juices, and soft drinks. The content of the composition of the present invention may be determined in a timely manner in the range of 0.1% to 70%, but it is preferable to determine the blending amount so that the above-mentioned guideline for daily intake can be ingested. .

  As feed, for example, grains such as corn, wheat, barley and rye, bran such as bran and rice bran, potatoes such as corn gluten meal and corn jam meal, skimmed milk powder, whey and fish meal It can be produced by blending animal feeds such as bone meal, yeasts such as beer yeast, calciums such as calcium phosphate and calcium carbonate, vitamins, fats and oils, amino acids, saccharides and the like. As a form of feed, it can be used for pet food, livestock feed, feed for cultured fish, and the like. The content of the composition may be determined in a timely range of 0.01 to 20%, so that a daily intake of 0.001 mg to 100 mg of carotenoid and 0.01 mg to 1 g of sphingolipid per kg of body weight is possible. It is preferable to add to feed raw materials.

  Examples of this experiment are described below. In addition, this invention does not limit the range by this Example. The measurement of cryptoxanthin used high performance liquid chromatography (HPLC). As an HPLC apparatus, LC-10A (manufactured by Shimadzu Corporation) was used, a Resolve C18 (φ3.9 × 150 mm, manufactured by Waters) column was connected, and a sample to which an equal amount of methanol was added was introduced. The mobile phase was analyzed with methanol: ethyl acetate = 7: 3, a column temperature of 30 ° C., a flow rate of 1.0 ml / min, and a detection wavelength of 450 nm.

  For the measurement of sphingolipid, an HPLC apparatus LC-10A connected with an Inertsil SIL100A column (manufactured by GL Sciences Inc.) was used and detected with a light scattering detector (500 ELSD, manufactured by ALLTECH). As a mobile phase, a linear gradient with two liquids of chloroform and chloroform / methanol (1/1) was used, and measurement was performed at a flow rate of 1.0 ml / min and 37 ° C.

Example 1
Juice of Satsuma mandarin was squeezed with an inline squeezer and filtered with a finisher (0.5 mm screen), and light centrifugation (2,500 × g · min) was performed with a centrifuge (Alpha Laval BRPX417) ). The supernatant portion of the light centrifugation was subjected to heavy centrifugation (4,000 × g · min) with the same centrifuge, and the precipitate was recovered. Extraction was carried out by adding 4.5 L of ethanol to 1.2 kg of powder that was freeze-dried and pulverized with a pulverizer, 8 kg of 8 kg of the juice residue (mandarin orange juice fine pulp, water content 85%). After filtration, the ethanol fraction was collected and ethanol was distilled off using an evaporator. Then, 300 ml of water was added and stirred, followed by filtration to collect a water-insoluble component, and the same operation was repeated twice. As a result, 10 g of Wenzhou mandarin orange extract was obtained. This extract contained 4% by weight of cryptoxanthin and 20% by weight of sphingolipid.

Test example 1
An extract containing the cryptoxanthin prepared in Example 1 and the sphingolipid was mixed with an emulsifier to 26 moderately obese men (average age 40.7 years old) having a BMI of 25 to 30 to prepare an emulsified preparation. This emulsified preparation was processed into juice, and two juices (12.5 mg / day as an extract (β-cryptoxanthin 0.5 mg / day, sphingolipid 2.5 mg / day)) were ingested for 12 weeks. The test was carried out in a double-blind manner with 13 persons in each group as a juice (Comparative Example 1) containing no extract as a comparison. CT scan imaging was performed mainly at the L4 / L5 intervertebral disc cross section, and the visceral fat measurement software Fat Scan Ver3.0 (N2 System) was used to measure the abdominal fat area, subcutaneous fat area, and total fat area. In addition, body weight, waist, and neutral fat were also measured. As a result, significant differences were observed with respect to visceral fat area, subcutaneous fat area, total fat area, body weight, waist, and neutral fat compared to Comparative Example 1 or before ingestion. Each result is shown in FIG. 1, FIG. 2, FIG. 3, FIG. 4, FIG. Obviously, a reducing action was observed for body fat such as visceral fat and subcutaneous fat and neutral fat. In addition, as a result of the reduction in body fat, a reduction in body weight and waist was also observed.

Test example 2
KK-Ay / TacJc1 female mice established as type 2 diabetes model animals (6 weeks old: Nippon Claire Co., Ltd.) are divided into 6 groups per group, and β-cryptoxanthin (manufactured by Shikoku Hachishu Pharmaceutical Co., Ltd., (Purity 95%) and glucosylceramide (unitika konjac ceramide, purity 3%) were mixed with high-fat feed at a rate of 0.2% and 4%, respectively, and allowed to freely ingest with water. As a control group, a high-fat feed (Comparative Example 2) to which nothing is added, a high-fat feed to which 0.2% of β-cryptoxanthin alone is added (Comparative Example 3), and a high-fat diet to which only 4% of glucosylceramide is added ( Comparative example 4) was given to mice and the test was carried out.

  Administration was carried out for 4 weeks, and body weight and triglyceride amount were measured before administration and on 7, 14, 21, and 28 days after administration. Further, the mice were sacrificed on the 29th day from the start of administration, and visceral fat, that is, white adipose tissue of the abdominal cavity was collected, and the weights were compared. The respective results are shown in FIG. 7, FIG. 8, and FIG.

  The body weight in the Test Example 2 group tended to decrease from the 7th day compared to Comparative Example 2 and Comparative Example 4, and decreased significantly on the 14th, 21st and 28th days (FIG. 7). It was confirmed that the amount of triglyceride also decreased significantly in Test Example 2 after the 21st day (FIG. 8). In addition, the abdominal fat tissue weight was significantly decreased in Test Example 2 and Comparative Example 3 as compared with Comparative Example 2 and Comparative Example 4. Furthermore, it was confirmed that the adipose tissue weight of Test Example 2 was significantly smaller than that of Comparative Example 3 (FIG. 9).

  As described above, the composition of the present invention has a significantly stronger body fat and neutral fat reducing effect than when it is combined with carotenoids and sphingolipids at the same time. It was confirmed that the composition was high in oral administration.

It is a figure which shows transition of the abdominal fat area of a L4 / L5 crossing part. It is a figure which shows transition of the subcutaneous fat area of L4 / L5 crossing part. It is a figure which shows transition of the total fat area of a L4 / L5 crossing part. It is a figure which shows transition of a body weight. It is a figure which shows transition of a waist. It is a figure which shows transition of neutral fat (TG). It is a figure which shows transition of a body weight. It is a figure which shows transition of the amount of serum triglycerides. It is a figure which shows transition of the visceral fat weight.

Claims (3)

For body fat reduction, comprising an extract containing cryptoxanthin and sphingolipid, obtained by extracting the pulp obtained by squeezing juice after squeezing mandarin orange with an organic solvent Oral medicine. The oral medicine for reducing body fat according to claim 1, which is for reducing visceral fat or subcutaneous fat. The oral medicine for body fat reduction according to claim 1 or 2, wherein the organic solvent is ethanol.