JP2010254590A - Pancreatic lipase inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a pancreatic lipase inhibitor and a lipid absorption inhibitor which have a pancreatic lipase inhibiting function, can make a contribution to the prevention and improvement of obesity, have a high preference, hardly have the problem of side reactions even when continuously taken, are effective, and have high safety. <P>SOLUTION: The pancreatic lipase inhibitor and the lipid absorption inhibitor include an extract of a plant belonging to the genus Rubus (for example, boysenberry) as an active ingredient. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

Background of the Invention

FIELD OF THE INVENTION The present invention relates to a pancreatic lipase inhibitor that can effectively inhibit pancreatic lipase, which is a lipolytic enzyme, using an extract of a plant belonging to the genus Rubus L as an active ingredient. The present invention also relates to a lipid absorption inhibitor containing such an extract. Furthermore, this invention relates to the food-drinks or pharmaceutical composition containing a pancreatic lipase inhibitor or a lipid absorption inhibitor.

Background Art Metabolic syndrome, that is, visceral fat type obesity and a state in which multiple hyperglycemia, hypertension and hyperlipidemia are duplicated, is known to cause various diseases, maintaining the health and quality of life (QOL) of the people It is important to avoid it. In particular, prevention and improvement of obesity by visceral fat accumulation is important, and exercise therapy and diet therapy have been proposed and implemented.

  Among these, in diet therapy, weight loss is considered to be most effective, but it is often difficult to continue in daily life due to excessive stress due to dietary restrictions. For this reason, research and development of foods having a digestive absorption inhibitory effect are underway.

  Pancreatic lipase secreted from the pancreas is a digestive enzyme that breaks down fat (neutral fat) taken by meals into fatty acids and glycerin. These degradation products of pancreatic lipase are absorbed from the small intestine, re-synthesized, then form chylomicrons with phospholipids, cholesterol and proteins, secreted into the lymphatic vessels, and pass through the thoracic duct into the aorta . If the fat is excessive, re-synthesized fats will accumulate in the body. Therefore, by inhibiting the action of pancreatic lipase, it is expected to inhibit or inhibit fat digestion and absorption from the intestinal tract, thereby reducing blood lipids.

For this reason, a component having an inhibitory activity on pancreatic lipase has attracted attention and has been studied.
For example, “Orlistat” is known as an ethical drug that is approved and sold overseas mainly in Europe and the United States. Orlistat is said to exhibit an anti-obesity effect and an effect of improving hyperlipidemia by suppressing absorption of fat ingested by meals due to its pancreatic lipase inhibitory activity. However, it has been reported that there are side effects such as rapid constipation, oil / fatty stool, oily diarrhea, and constipation (Obesity Research, Vol. 7, No. 3, pp. 112-114, 2001).

  For this reason, there is a need for safe and effective pancreatic lipase inhibitors and lipid absorption inhibitors with few side effects. In view of this, plant materials having pancreatic lipase inhibitory activity have been studied.

  For example, extracts of fruits and their seeds (Meiji University Faculty of Agriculture report, 69, 15, 1985 (Non-patent document 1)), hesperidin of Wenzhou oranges (Biosci. Biotech. Biochem., 61, 102, 1997 (Non-patent document 2) )), Apple polyphenol-derived procyanidins (J. Agric. Food Chem., 55, 4604, 2007 (Non-Patent Document 3)), Kawaroketsume-derived tannins (JP-A-8-259557 (Patent Document 1)), tamarind seed coat The pancreatic lipase inhibitory activity of the derived proanthocyanidins (JP-A-9-291039 (Patent Document 2)) and spices and extracts thereof (JP-A 2001-120237 (Patent Document 3)) have been reported.

As described above, various plant materials having pancreatic lipase inhibitory activity are already known. However, raspberry acids in these prior art, particularly with respect to fruit and its extracts Boysenberry (Rubus loganobaccus), not described or suggested in any way.

  Boysenberry, unlike raspberry, blackberry and blueberry, is not well known in Japan, but is well known in Europe and America because of its unique flavor. Regarding the functionality of Boysenberry-derived components, for example, in the literature (Journal of Japanese Society for Dietary Life, Vol.16, No.1, pp.44-49, 2005 (non-patent document 4)), polyphenols such as anthocyanins and ellagic acid are used. It describes that it contains a lot of components that exhibit the antioxidative action. Here, it is expected that anthocyanins contained in Boysenberry fruits and leaves are expected to have an anti-aging effect, an action that protects cardiovascular function, an anti-diabetic action, and a liver damage-protecting action. Japanese Patent Laid-Open No. 2008-156306 (Patent Document 4) discloses that a Boysenberry fruit-derived component can exhibit a carcinogenesis-inhibiting action.

  However, these documents neither disclose nor suggest that Boysenberry has pancreatic lipase inhibitory activity or lipid absorption inhibitory effect.

  Japanese Patent No. 3967103 (Patent Document 5) discloses a lipase activity inhibitor useful for the treatment of acne, which includes a kobotan extract. Here, Boysenberry fruit is mentioned as one of many examples of plants for plant-derived additive components added to external preparations and cosmetics. However, here it is only suggested that the plant-derived additive component can be used for the purpose of skin protection, moisturizing effect, prevention of rough skin, fragrance, etc., and these additive components themselves have lipase inhibitory activity. There is no disclosure or suggestion about whether or not.

  Polyphenol is a general term for physiologically active substances including coumarin, isoflavone, flavonol, anthocyanin, and ellagic acid (Non-Patent Document 4), and is known to have an antioxidant effect. Moreover, it is known that polyphenol is contained in red wine, oolong tea, etc., polyphenol derived from oolong tea (Japanese Patent Laid-Open No. 2006-001909 (Patent Document 6)), apple polyphenol (Non-Patent Document 3), etc. In addition, it has been reported to have a lipase inhibitory activity.

  However, since the definition of a substance called polyphenol itself is an ambiguous term that includes many phenols, the activity of polyphenols may vary greatly depending on the plant species from which they are derived. For this reason, it cannot necessarily be said that the polyphenol derived from another plant has the activity which the polyphenol derived from a certain plant has. In addition, since such polyphenols originate from natural products, in order to obtain the desired effect, it may be necessary to consume a large amount of plant-derived components or extremely concentrated components. is there. In such cases, the bitterness and astringency may become too strong, the intake of caffeine etc. may be maximized, or the taste may be unacceptable for taste, which may be a problem for daily consumption. there were.

  As described above, the antiphenolic activity of polyphenols derived from plants belonging to the genus Rubus, particularly Boysenberry, has been reported, but there has been no disclosure or suggestion of pancreatic lipase inhibitory activity.

JP-A-8-259557 JP-A-9-291039 JP 2001-120237 A JP 2008-156306 A Japanese Patent No. 3967103 JP 2006-001909 A

Report of Faculty of Agriculture, Meiji University, 69, 15, 1985 Biosci. Biotech. Biochem., 61, 102, 1997 J. Agric. Food Chem., 55, 4604, 2007 Journal of Japanese Society of Food Life, Vol.16, No.1, pp.44-49, 2005

  The present inventors have now unexpectedly found that the fruit of Boysenberry, which is a plant belonging to the genus Rubus, and its extract have a strong inhibitory activity against pancreatic lipase. Furthermore, when the effect of fat on the body absorption was actually investigated using rats, it was found that lipid absorption was suppressed by Boysenberry fruit and its extract. The present invention is based on these findings.

  Therefore, the present invention has an inhibitory function of pancreatic lipase, can contribute to the prevention and improvement of obesity, has high palatability, has almost no fear of side effects even when used continuously, and is effective and highly safe, inhibiting pancreatic lipase It is an object of the present invention to provide an agent, a lipid absorption inhibitor, and a food and drink and a pharmaceutical composition containing them.

The pancreatic lipase inhibitor according to the present invention contains an extract of a plant belonging to the genus Rubus L as an active ingredient.

The lipid absorption inhibitor according to the present invention contains the above extract of the present invention, that is, an extract of a plant belonging to the genus Rubus L as an active ingredient.

According to one preferable aspect of the present invention, in the above-mentioned agent, the plant belonging to the genus Rubus is Boysenberry ( Rubus loganobaccus ).

  According to one more preferable aspect of the present invention, in the above-mentioned agent, the extract is obtained by extraction from plant fruit, fruit juice, or plant skin and seed.

  According to another preferred embodiment of the present invention, in the agent described above, the extract is a water extract, an organic solvent extract or a water-containing organic solvent extract.

  According to another more preferable aspect of the present invention, in the aforementioned agent, the extract is a Boysenberry hydrous organic solvent extract.

  According to another aspect of the present invention, there is provided a food or drink comprising the pancreatic lipase inhibitor according to the present invention. According to still another aspect of the present invention, there is provided a food or drink comprising the lipid absorption inhibitor according to the present invention.

  According to still another aspect of the present invention, there is provided a method for producing a food or drink, comprising adding the pancreatic lipase inhibitor according to the present invention or the lipid absorption inhibitor according to the present invention to a material component of the food or drink. Is done.

  Furthermore, the pharmaceutical composition according to the present invention is for inhibiting pancreatic lipase activity in the body, and comprises the above-described extract of the present invention as an active ingredient.

  According to one preferred embodiment of the present invention, the pharmaceutical composition is used for the treatment, prevention or amelioration of a disease or condition that can be treated, prevented or ameliorated by inhibiting pancreatic lipase. More preferably, the disease or condition is obesity or hyperlipidemia.

  According to one preferred embodiment of the present invention, the pharmaceutical composition is used for lowering lipids in the body.

  According to the pancreatic lipase inhibitor and lipid absorption inhibitor of the present invention, pancreatic lipase activity can be effectively and safely inhibited, fat digestion and absorption in the body can be suppressed, and lipids can be further inhibited. Since the absorption into the body can be effectively suppressed, blood lipids can be reduced. Therefore, the pancreatic lipase inhibitor and lipid absorption inhibitor according to the present invention are extremely useful for the prevention and improvement of obesity. Moreover, since the pancreatic lipase inhibitor and lipid absorption inhibitor according to the present invention uses an extract of a plant belonging to the genus Rubus that is also used as a food (for example, Boysenberry), it has high palatability and can be used continuously. There is almost no risk of side effects and the safety is extremely high. In addition, according to the present invention, since foods and drinks and pharmaceutical compositions containing a pancreatic lipase inhibitor and a lipid absorption inhibitor can be provided, ingesting these inhibits pancreatic lipase activity in the body, digestion of fat, Absorption can be suppressed, absorption of lipids into the body can be effectively suppressed, and blood lipids can be reduced. Furthermore, the pancreatic lipase inhibitor, lipid absorption inhibitor, food and drink, and pharmaceutical composition according to the present invention are effective for the prevention / improvement of metabolic syndrome caused by pancreatic lipase inhibitory effect in addition to the treatment, prevention, and improvement of obesity.

It is a graph which shows the result of Test example 2 of an example. It is a graph which shows the result of Test example 2 of an example.

Detailed description of the invention

Active ingredient The pancreatic lipase inhibitor or lipid absorption inhibitor according to the present invention comprises, as described above, an extract of a plant belonging to the genus Rubus L as an active ingredient.

Here, the plant belonging to the genus Rubus is a plant belonging to the family Rosaceae, and is characterized in that it produces fruit that gathers small fruits. There are many wild species and cultivars of plants belonging to the genus Rubus, and well-known ones include, for example, blackberry ( Rubus ursinus ), raspberry ( Rubus ideaus ), boysenberry ( Rubus loganobaccus ), etc. It is done. In many cases, these plants belonging to the genus Rubus are eaten with their fruits intact or processed into fruit juice or jam. According to a preferred embodiment of the present invention, the plant belonging to the genus Rubus is one selected from the group consisting of blackberry, raspberry, and boysenberry, more preferably boysenberry.

  Boysenberry is considered to be a variety made by crossing blackberry, raspberry, and loganberry, and its fruit is characterized by a brilliant purple color, rich aroma, and a good sweetness and acidity. Boysenberry is rich in calcium, iron, zinc, vitamin A, folic acid, niacin, vitamin C, and contains nutrients in a well-balanced manner.

  In the present invention, the “extract” is obtained by extracting from the fruits, fruit juices, pericarps and seeds of plants belonging to the genus Rubus, and the extract, the diluted solution, the concentrated solution, the extract or the dried product or the dried powder thereof is used. means. The operation for performing extraction here includes squeezing extraction, solvent extraction using water or an organic solvent, and the like, and further treatments such as adsorption, filtration, and centrifugation may be performed as necessary. In the present invention, the extraction operation is preferably a solvent extraction operation using water or an organic solvent.

  Here, the organic solvent may be either a water-soluble or water-insoluble organic solvent as long as an extract having a desired effect can be extracted. When the organic solvent is water-soluble (water-miscible), it may be used by mixing with water, and such a water-containing organic solvent can also be used. Therefore, for example, when the organic solvent is a water-soluble alcohol solvent, a hydrous alcohol solvent extract obtained by extraction using an alcohol solvent containing water at a certain ratio is also included in the extract. The

  Specific examples of the organic solvent include methanol, ethanol, isopropanol, acetonitrile, butanol, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, and methyl ethyl ketone. Two or more of these may be mixed and used for extraction, and if possible, mixed with water.

  From the viewpoint of adapting to foods and drinks or pharmaceutical compositions, it is preferable to use water alone or a mixture of water and ethanol as the extraction solvent from the viewpoint of safety in view of the remaining solvent.

  Thus, according to one preferred embodiment of the present invention, the extract is a water extract, an organic solvent extract or a hydrous organic solvent extract. More preferred is a hydrous organic solvent extract, and still more preferred is a hydrous ethanol extract.

  According to a preferred embodiment of the present invention, the fruit, fruit juice, pericarp and seed of a plant belonging to the genus Rubus are treated with water, an organic solvent or a water-containing organic solvent, and this is treated with an adsorbent substance (preferably polyphenol). Substance (for example, a synthetic adsorbent, activated carbon, ion exchange resin, polyvinyl pyrrolidone, diatomaceous earth, or a combination thereof) and adsorbed, and then eluted using a solvent to form an effluent. Accordingly, the effluent can be concentrated or dried and subjected to a purification treatment such as filtration to obtain an extract as an active ingredient.

  The method for producing an active ingredient in the present invention will be further described with specific examples. The juice of a plant belonging to the genus Rubus is diluted with water and applied to a synthetic adsorbent, for example, eluted with 80% ethanol. The effluent was then concentrated under reduced pressure to distill off the solvent, followed by centrifugation and filtration to obtain a clarified extract. Ethanol extract) can be obtained.

  The obtained extract is usually easily water-soluble and can maintain stable properties as an aqueous solution. For this reason, it is excellent in miscibility with the component which can be normally added to food-drinks and a pharmaceutical composition. Moreover, since it is a component contained in food, it is nontoxic and excellent in safety. Therefore, pancreatic lipase inhibitors or lipid absorption inhibitors containing these as active ingredients are excellent in safety, have almost no side effects, and can be easily taken in by appropriately incorporating them into daily eating habits.

  The pancreatic lipase inhibitor or lipid absorption inhibitor according to the present invention can be used alone, but can be added as an additive to various compositions such as foods and drinks, pharmaceuticals, etc. A composition having an absorption suppressing effect) can be obtained. The obtained composition can be effectively used for the prevention and improvement of metabolic syndrome caused by the pancreatic lipase inhibitory effect in addition to the treatment and prevention of obesity.

  According to another preferred embodiment of the present invention, a fruit juice of a plant belonging to the genus Rubus can be used as an active ingredient. More preferably, at this time, the extract can be further added to the fruit juice.

The active ingredient in the present invention is an extract as described above, and it is predicted that the main component is a polyphenol derived from a plant belonging to the genus Rubus or a polyphenol derived from Boysenberry. It is not limited. On the other hand, depending on the type and part of the plant used, the extraction conditions, and the like, it is considered that the amount of active ingredient (amount of extract) that can be expected to have a desired effect may change.
Therefore, in determining the amount of active ingredient to be used, the known method for measuring the total amount of polyphenols is used to identify the effective amount of the extract obtained as a polyphenol amount and extract it as the active ingredient to be used. It can be determined as physical quantity. Such a known method for measuring the total amount of polyphenols is the Folin-Denis method (O. FOLIN and W. DENIS: J. Biol. Chem., 12, 239, 1912, or “Food Function Research Method” (Kotsumitsu). (Published, 2000), 318-319), Folin-Ciocalteu method (VL SINGLETON and JA ROSSI: Amer. J. Enol. And Vitic., 16, 144, 1965).

  For example, in the case of Boysenberry extract (for example, one produced in Test Example 1), the total polyphenol amount in 1 g of the extract was 647 mg / g by the Folin-Ciocalteu method (in terms of gallic acid). Therefore, it is possible to specify the amount of extract as an active ingredient to be used by calculating the desired intake amount in terms of polyphenol amount.

Use The plant extract, which is an active ingredient according to the present invention, has pancreatic lipase inhibitory activity (Test Example 1 in Examples described later), and has an effect of suppressing lipid absorption into the body (in Examples described later). Test Example 2).

  Here, the inhibitory activity of pancreatic lipase means an action capable of inhibiting the lipolytic activity of pancreatic lipase contained in the pancreatic juice of mammals including humans. In general, lipases are widely distributed from animals and plants to microorganisms, are localized in tissues and organs in the living body, and play a role as a catalyst in the decomposition reaction of fats and oils. However, as can be seen from the extremely diverse physiological functions of the tissues and organs constituting each organism, the molecular structures and the existence states of lipids in these tissues and organs are also diverse. For this reason, even if they belong to the same category of lipases, their effects on substrates and enzymatic properties are not uniform. Therefore, pancreatic lipase can also be said to be a specific embodiment among many lipase groups. In this invention, it is based on that an active ingredient shows effective inhibitory activity with respect to pancreatic lipase.

  Here, the lipid absorption inhibitory effect refers to an action that can suppress the absorption of fat ingested by the body into the body, thereby treating obesity, prevention, improvement, treatment of metabolic syndrome, Prevention and improvement are carried out.

  As described above, when pancreatic lipase activity is inhibited, digestion of ingested fat and absorption from the intestinal tract are inhibited or suppressed, resulting in a decrease in blood fat. That is, by inhibiting pancreatic lipase, the absorption of lipids in the body is suppressed, and the action of lowering lipids in the body is exhibited. Such a relationship is well known, and for example, as textbook literature, (i) “New Clinical Nutrition” (Nanedo, 3rd edition, 1999), pages 73-77 and 95- 101; (ii) "Color illustration: Normal structure and function of the human body II Gastrointestinal tract" (published by Nippon Medical Newspaper, 2000), page 58; and "Studies on functional substances in food and obesity", Japan The Journal of Nutrition / Food Science, Vol. 54, No. 1, pages 35-40, 2001), and the above relationship is clearly shown. Therefore, since the active ingredient according to the present invention has pancreatic lipase inhibitory activity and has also been confirmed to actually suppress lipid absorption into the body, the active ingredient according to the present invention can treat, prevent and improve obesity. In addition, it is clear that it is effective in preventing and improving metabolic syndrome caused by pancreatic lipase inhibitory activity.

  Therefore, the active ingredient according to the present invention can be used for treatment, prevention or amelioration of a disease or condition that can be treated, prevented or ameliorated by inhibiting pancreatic lipase. Here, diseases or conditions that can be treated, prevented, or improved by inhibiting pancreatic lipase include obesity, insulin resistance, postprandial hyperglycemia, hypertension, hyperlipidemia, fatty liver, diabetes, cerebrovascular disorder, ischemic heart Disease etc. are mentioned. Preferably, the disease or condition is obesity or hyperlipidemia. According to a further preferred embodiment of the present invention, the active ingredient of the present invention is used for lowering lipids in the body, particularly for suppressing lipid absorption.

  In the present specification, “treatment, prevention or amelioration” of a disease or condition is used in a sense including regulation, delay of progression, alleviation, prevention of onset, prevention of recurrence, suppression of the disease or condition.

  According to another aspect of the present invention, there is provided a method for inhibiting pancreatic lipase comprising administering to a mammal an effective amount of an extract which is an active ingredient of the present invention. Here, the “effective amount” is an amount sufficient to exert pancreatic lipase inhibitory activity in a desired region in the body by administration.

  According to another aspect of the present invention, treatment, prevention or improvement is achieved by inhibiting pancreatic lipase, comprising administering to a mammal a therapeutically effective amount of an active ingredient which is an active ingredient of the present invention. Methods of treating, preventing or ameliorating possible diseases or conditions are provided. As used herein, “therapeutically effective amount” is an amount sufficient to ameliorate one or more symptoms usually associated with the disease or condition for which treatment is desired. When referring to prophylactic use, the term means an amount sufficient to prevent or delay the onset of the disease or condition.

Food / beverage products and pharmaceutical composition As described above, according to the present invention, a food / beverage product comprising the pancreatic lipase inhibitor according to the present invention is provided. Moreover, according to this invention, the food / beverage products which comprise the lipid absorption inhibitor by this invention are provided. That is, these foods and drinks contain an extract of a plant belonging to the genus Rubus as an active ingredient.

  Such a food or drink is produced, for example, by a production method comprising adding the pancreatic lipase inhibitor according to the present invention, the lipid absorption inhibitor according to the present invention, or the extract to the material component of the food or drink. be able to.

  Moreover, according to this invention, as mentioned above, it is for inhibiting pancreatic lipase activity in a body, Comprising: The pharmaceutical composition which contains the extract in above-mentioned this invention as an active ingredient is provided.

  As used herein, “comprising as an active ingredient” may contain a physiologically acceptable carrier according to the desired product form, and may contain other auxiliary ingredients that can be used in combination. It means to include.

  In the present invention, the food and drink is other than the pharmaceutical composition and is not particularly limited as long as it is a form that can be taken orally, such as a solution, suspension, emulsion, powder, or solid molded product. Specifically, for example, instant noodles, retort food, canned food, microwave food, instant soup / miso soup, freeze-dried food, etc .; soft drink, fruit juice drink, vegetable drink, soy milk drink, coffee drink, tea Beverages such as beverages, powdered beverages, concentrated beverages, alcoholic beverages; bread products such as bread, pasta, noodles, cake mixes, fried flour, bread crumbs; rice cakes, caramel, chewing gum, chocolate, cookies, biscuits, cakes, pies, Confectionery such as snacks, crackers, Japanese confectionery, dessert confectionery; sauces, tomato processing seasonings, flavor seasonings, cooking mixes, sauces, dressings, soups, curry and stew ingredients; processed fats and oils, Fats and oils such as butter, margarine, mayonnaise; milk drinks, yogurts, lactic acid bacteria drinks, ice creams, Dairy products such as reams; processed fish products such as egg processed products, fish ham and sausages, fish paste products; processed livestock products such as livestock ham and sausages; agricultural products such as canned agricultural products, jams and marmalades, pickles, boiled beans and cereals Processed products; frozen foods and the like.

  Foods and drinks are classified as health foods, foods for specified health use, functional foods for the sick, foods for the sick, infant formula, powdered milk for pregnant or lactating women, or foods and drinks with a label for reducing disease risk. Are also included.

  According to another aspect of the present invention, there is provided a food / beverage product comprising an effective amount of the extract, which displays a function of treating, preventing or improving a disease or condition that can be treated, prevented or ameliorated by inhibiting pancreatic lipase. Food and drink may be provided. According to still another aspect of the present invention, there can be provided a food or drink comprising an effective amount of the extract, which is displayed with pancreatic lipase inhibitory ability or lipid absorption inhibitory ability.

  Therefore, the food and drink according to the present invention can be provided as, for example, a food suitable for consumers who expect improvement or alleviation of obesity, a food suitable for suppressing the rise in lipids, that is, a so-called food for specified health use. it can. The food for specified health use here refers to a disease or condition that can be treated, prevented or ameliorated by inhibiting pancreatic lipase, obesity prevention, improvement, condition alleviation, metabolic syndrome treatment, prevention or improvement, etc. When manufacturing or selling foods and drinks, it can refer to foods and drinks that may be subject to some legal restrictions in each country from the viewpoint of health. Such a food or drink may be a food or drink that indicates that the food or drink may reduce a disease risk, that is, a food or drink with a disease risk reduction display. Here, the disease risk reduction label is a label for foods and drinks that may reduce the disease risk, and is based on a standard established by the FAO / WHO Joint Food Standards Committee (Codex Committee) or its With reference to the standard, it can be a defined display or an approved display.

  In the present invention, the pharmaceutical composition is prepared as an oral preparation or a parenteral preparation according to a conventional method using additives that are acceptable for formulation. For oral preparations, take the form of solid preparations such as tablets, powders, fine granules, granules, capsules, pills, sustained-release preparations, and liquid preparations such as solutions, suspensions, and emulsions. Can do. In the case of a parenteral preparation, it can take the form of an injection or a suppository. From the viewpoint of simplicity, an oral preparation is preferable. Additives that are acceptable for formulation include, for example, excipients, stabilizers, preservatives, wetting agents, emulsifiers, lubricants, sweeteners, colorants, fragrances, buffers, antioxidants, pH Examples thereof include regulators.

  The content of the active ingredient of pancreatic lipase inhibitor in a composition such as a food or drink or a pharmaceutical composition is difficult to define uniformly depending on the type and form of the composition, the purpose of prevention / improvement, etc. Adults (body weight 60 kg) per day as Boysenberry extract (for example, those produced in Test Example 1) are about 50 to 1000 mg, preferably about 200 to 400 mg. What is necessary is just to set the content in a composition suitably in consideration of the intake of the composition per day so that the required intake of the active ingredient per day can be taken.

  The present invention will be described in detail by the following examples, but the present invention is not limited thereto.

Test example 1
a) Preparation of sample solution 200 g of Boysenberry concentrated fruit juice (8-fold concentrated fruit juice) was applied to 700 ml of a synthetic adsorption resin (trade name: XAD-7HP, Organo Corporation). Next, the synthetic adsorption resin was washed with 1000 ml of deionized water, and then the adsorbate in the resin was eluted and collected using 800 ml of 80% ethanol to obtain an effluent. This was concentrated to 80 ml at about 38 ° C. in an evaporator, and then freeze-dried to obtain Boysenberry extract (powder product) (Boysenberry polyphenol powder product). A sample solution (sample solution of Boysenberry extract) was prepared using 100% ethanol so that the final concentration of the extract was 8 mg / ml.
In addition, the Boysenberry concentrated fruit juice freeze-dried as it was was used as Boysenberry juice powder, and this was similarly adjusted with ethanol so that the final concentration of the extract was 8 mg / ml, and the sample solution (Boysen Berry juice powder sample).
As a positive control, a known pharmaceutical orlistat (Orlistat (manufactured by SIGMA)) known to have pancreatic lipase inhibitory activity was used.

b) Preparation of pancreatic lipase solution and substrate solution Pancreatic lipase activity was calculated by measuring the amount of oleic acid released from triolein. Porcine pancreatic lipase (manufactured by SIGMA) was prepared as a pancreatic lipase enzyme, and this was dissolved in a phosphate buffer and adjusted to a concentration of 50 μg / ml, and this was used as a pancreatic lipase solution.
The phosphate buffer used here was prepared by adjusting a 0.2 M sodium dihydrogen phosphate (dihydrate) aqueous solution and a 0.2 M disodium hydrogen phosphate aqueous solution, respectively, and collecting 39 ml and 61 ml, respectively. Deionized water was added to obtain 200 ml.

  Next, 80 mg of triolein, 10 mg of lecithin, and 5 mg of sodium taurocholate are weighed into a test tube, 9 ml of the buffer solution is added thereto, and subjected to ultrasonic treatment for 10 minutes to obtain a lipid emulsion solution, which is used as a substrate solution. used.

c) Pancreatic lipase inhibitory activity test sample solution 0.1 ml of pancreatic lipase solution (final concentration 10 μg / ml) was added and mixed, incubated for 5 minutes at room temperature, and then 0.1 ml of substrate solution was added thereto. The mixture was mixed and reacted for 30 minutes at a temperature of 37 ° C.
After the reaction, 1 ml of chloroform was added and mixed to stop the reaction. This was centrifuged at 3000 rpm for 10 minutes, and a dried product was obtained from the upper chloroform layer. To this, 40 μl of propanol was added and dissolved, and then diluted 10 times to obtain a reaction solution.
The concentration of free fatty acid in the reaction solution was measured for absorbance at 550 nm with a microplate reader using an enzyme method measurement kit NEFA C-Test (manufactured by Wako Pure Chemical Industries, Ltd.). From the obtained value, the free fatty acid concentration of each sample solution was calculated based on a standard curve of oleic acid prepared in advance.

d) Calculation of pancreatic lipase inhibition rate As a control (control), the free fatty acid concentration using ethanol was set to 100%, and the inhibition rate of pancreatic lipase was calculated from the free fatty acid concentration of the sample solution according to the following formula.
Moreover, the inhibitory concentration of the sample which halves the activity of the control was determined, and this was expressed as IC ₅₀ (mg / ml).

[Inhibition rate of pancreatic lipase (%)]
= (1-Free fatty acid concentration of sample solution ÷ Free fatty acid concentration of control) × 100

The results were as shown in Table 1 and Table 2 below.
Table 1 shows the pancreatic lipase inhibitory activity of Boysenberry extract according to the present invention, Table 2, expressed as IC _50.

From the results, it was found that the Boysenberry extract showed high pancreatic lipase inhibitory activity of 78.3% at a final concentration of 8 mg / ml. For this reason, it was confirmed that an inhibitory effect equivalent to or higher than that of existing orlistat can be obtained by adjusting the concentration.
IC ₅₀ was 3.6 mg / ml for orlistat and 5.0 mg / ml for Boysenberry extract. Although not as much as orlistat, it was found that Boysenberry extract can also exhibit excellent inhibitory activity. Anti-obesity drugs containing orlistat as an active ingredient are known to have side effects, so using Boysenberry extract can be expected to prevent and suppress obesity safely and effectively without side effects. all right.

Test example 2
a) Preparation of sample and oil / fat emulsion In the same manner as in item a) of Test Example 1, Boysenberry extract (powder product) (Boysenberry polyphenol powder product) was obtained from Boysenberry concentrated fruit juice.
Further, as in the case of item a) of Test Example 1, a Boysenberry juice concentrate was prepared by freeze-drying the Boysenberry juice concentrate as a Boysenberry juice powder.
100 mg of cholic acid was dissolved in 5 ml of corn oil, 5 ml of distilled water was further added, and sonication was performed for 10 minutes to obtain an oil and fat emulsion.

b) For the administration test to rats, 9-week-old Wistar male rats obtained from Japan SLC Co., Ltd. were used. One week before the start of the test, the rats were fed a normal diet and preliminarily reared. Each group was divided into 3 groups so that the average body weight was constant.
After fasting for 20 hours, an oil and fat emulsion (5 ml / kg-body weight) and a sample solution (5 ml / kg-body weight) were collected in a syringe and mixed, and then administered orally. At this time, the concentration in the sample solution was adjusted so as to satisfy the following dosage conditions for each of the three groups to which the rats were distributed.
(i) Control group (control group): a group that did not contain Boysenberry extract and was administered only with an oil emulsion;
(ii) BJ group: a group in which Boysenberry juice powder was used, and this was administered to 100 mg / kg body weight in terms of Boysenberry polyphenol, and
(iii) BP500 group: A group in which a Boysenberry extract powder product was used, and this was administered so as to give 500 mg / kg body weight in terms of Boysenberry polyphenol.

  Blood was collected from the tail vein of rats before administration of the mixture containing the oil emulsion and 60, 120, 180, and 240 minutes after administration. The collected blood was stored on ice and then centrifuged at 13,000 rpm for 5 minutes to obtain serum.

In addition, in the above Boysenberry polyphenol amount conversion, Boysenberry extract (powder product) and Boysenberry fruit juice powder, in advance, by measuring the total polyphenol amount by the Folin-Ciocalteu method (gallic acid conversion), Conversion was performed based on this. Specifically, the total amount of Boysenberry polyphenol per gram of each was as follows:
Boysenberry extract (powder product) 647 mg / g,
Boysenberry juice powder 25 mg / g.

c) Measurement of blood triglyceride concentration The serum triglyceride concentration obtained was measured for absorbance at 600 nm with a microplate reader using an enzyme assay kit triglyceride E-test (manufactured by Wako Pure Chemical Industries, Ltd.). Was measured. From the obtained value, the blood neutral fat concentration of each individual was calculated based on a standard curve of a mixed liquid of glycerin and triolein prepared in advance.

  The results were as shown in FIG. 1 and FIG. The figure shows the amount of blood neutral fat increased after fat and oil administration.

  From the results, in the BP500 group to which the sample containing Boysenberry extract was administered, a strong suppression was observed against the increase in blood neutral fat concentration.

Production Examples The prescriptions are shown below as examples of producing foods and drinks and pharmaceutical compositions according to the present invention.

Claims (13)

Pancreatic lipase inhibitor comprising an extract of a plant belonging to the genus Rubus L as an active ingredient. The pancreatic lipase inhibitor according to claim 1, wherein the plant belonging to the genus Rubus is Boysenberry ( Rubus loganobaccus ).   The pancreatic lipase inhibitor according to claim 1 or 2, wherein the extract is obtained by extraction from plant fruit, fruit juice, or plant skin and seed.   The pancreatic lipase inhibitor according to any one of claims 1 to 3, wherein the extract is a water extract, an organic solvent extract or a water-containing organic solvent extract.   The pancreatic lipase inhibitor according to any one of claims 1 to 4, wherein the extract is a water-containing organic solvent extract of Boysenberry.   The lipid absorption inhibitor which uses the extract as described in any one of Claims 1-5 as an active ingredient.   A food or drink comprising the pancreatic lipase inhibitor according to any one of claims 1 to 5.   A food or drink comprising the lipid absorption inhibitor according to claim 6.   The manufacturing method of food-drinks which comprises adding the pancreatic lipase inhibitor as described in any one of Claims 1-5, or the lipid absorption inhibitor of Claim 6 to the material component of food-drinks. .   A pharmaceutical composition for inhibiting pancreatic lipase activity in the body, comprising the extract according to any one of claims 1 to 5 as an active ingredient.   The pharmaceutical composition according to claim 10, which is used for treatment, prevention or amelioration of a disease or condition which can be treated, prevented or ameliorated by inhibiting pancreatic lipase.   The pharmaceutical composition according to claim 11, wherein the disease or condition is obesity or hyperlipidemia.   The pharmaceutical composition according to any one of claims 10 to 12, which is used for lowering lipids in the body.

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