JP2010248212A - Glycation inhibitor and application thereof - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a glycation inhibitor generating no side reaction, which is usable as an additive for food, a prophylactic and therapeutic agent for tissue disorders, food and drink, and an external preparation. <P>SOLUTION: The glycation inhibitor contains at least one extract obtained by extracting a plant using an organic solvent or a combined liquid of an organic solvent and water as the extracting solvent, the plant selected from Agrimonia eupatoria, Hamamelis virginiana, Rubus suavissimus, Alchemilla vulgaris, Camellia sinensis, Salacia oblonga, Paulinia cupana and the like. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  TECHNICAL FIELD The present invention relates to a highly safe glycation inhibitor having a plant extract or a component thereof as an active ingredient and having effects of preventing, treating, and improving various tissue disorders caused by glycation, and use thereof.

  In recent years, factors that promote aging are increasing due to deterioration of environmental conditions, changes in lifestyle, and increased stress accompanying complications in social life. On the other hand, however, the average life expectancy of Japanese people has increased, the proportion of elderly people has increased, and the importance of living in a youthful and healthy manner has been reviewed. Therefore, the suppression of the progress of aging is also a place where everyone is waiting.

  Various factors are involved in the progress of aging, and one of the factors is a non-enzymatic protein saccharification reaction. This is a reaction that brings about an obstacle to the original function of a protein by binding of a reducing sugar and an amino acid or protein, and is called glycation (browning reaction). Proteins are responsible for various physiological functions in the living body, and glycated proteins cannot exert their original functions and are thought to cause various functional disorders. The initial reactant of this glycation is a protein-sugar conjugate called an Amadori compound, and this reaction is characterized by fluorescence, browning, intramolecular and intermolecular cross-linking through dehydration and polymerization. It becomes advanced glycation end products (AGE).

  These AGEs accumulate in the living body with aging, and various tissue disorders recognized by aging, for example, hardening of connective tissues such as skin, blood vessel walls, joints, and lungs (for example, see Non-Patent Documents 1 to 4). It has been suggested that it may cause skin thickening and wrinkle formation, arteriosclerosis, arthritis, cardiopulmonary hypofunction and the like. Moreover, it is thought that it is related also to the cataract (for example, refer nonpatent literature 5). In addition, it is estimated that glycation not only causes tissue damage due to aging but also participates in diseases such as sunlight elastic fibrosis, Alzheimer's disease, and diabetic complications. For example, carboxymethyllysine (CML), a type of AGE, has been detected in the skin tissue of sunlight elastic fibrosis, suggesting that it is involved in lesions such as deep wrinkle accumulation of CML and hardening of skin tissue. (For example, see Non-Patent Document 6). It has also been suggested that AGE is involved in the accumulation of β-amyloid peptide in the brain in Alzheimer's disease (see, for example, Non-Patent Document 7).

  Furthermore, it has also been suggested that accumulation of AGE is related to nephropathy and retinopathy in diabetes (see, for example, Non-Patent Document 8). Aminoguanidine is known as a drug that inhibits the glycation that is the source of such AGE accumulation. This compound has been confirmed in model animals to be effective for diabetic complications such as retinopathy, renal disorders, and neuropathy (see, for example, Non-Patent Document 9), and is clinically tested in the United States as a therapeutic agent for diabetic complications. Is underway. However, side effects such as headache and nausea have been reported by taking aminoguanidine, and it is necessary to take it under the guidance of a doctor.

Bai P. et al., Conn Tissue Res 1992; 28: 1-12 Sakata N. et al., Atherosclerosis 1995; 116: 63-75. Verzijl N. et al., Biochem J 2000; 350 Pt 2: 381-7. Bellmunt MJ. Et al., Lung 1995; 173: 177-85. Prabhakaram M. et al., Mech Ageing Dev 1996; 91 (1): 65-78 Mizutari K. et al., J Invest Dermatol 1997; 108 (5): 797-802 Vitek MP. Et al., Proc Natl Acad Sci USA 1994; 91 (11): 4766-7 0 McCance DR. Et al., J Clin Invest 1993; 91: 2470-8 Brownlee M., Diabetes Care 1992; 15: 1835-43.

  Accordingly, an object of the present invention is to provide a highly safe glycation inhibitor having a high glycation inhibitory effect and causing no side effects.

  In view of such circumstances, the present inventors have conducted intensive research to solve the above problems, and as a result, found that an extract of a specific plant has a high glycation-inhibiting effect and completed the present invention.

  That is, the present invention includes agrimony, ashwagandha, anise, amacha candy, sweet candy, ginko, fennel, witch hazel, turmeric, echinacea, elephant, elder, elba mate, plantain, olive, oregano, orange, cocoon, oyster , Guarana, ginseng, chrysanthemum, gymnema, cat's claw, catnip, killaway, goldfish, guava, cucumber, crispness, cumixtin, black soybeans, wrinkles, cinnamon, ketsumeishi, bitter gourd, cola nuts, gokahi, gotsukora, coriander, comfrey , Saximas Sou, Saflower, Salacia Oblonga, Hawthorn, Sancychin carrot, Shiso, Juyaku, Juniper berry, Ginger, Cedar, Sugina, Stevia, Spearmint, Sage, Savory, Sosa , Sen, ▲ investigation ▼, thyme, tahibo, dandelion, chervil, chanoki, chabotokeiso, clove, daisy, dill, eucommia, nutmeg, nanban, nettle, hibiscus, white moth, white snake tongue, hagoromoso, basil, Lotus, Half-branch, Heath, Peach, Hyssop, Biwa, Fever-fu, Fukugi, Wisteria, Plantago sylum, Blueberry, Blue mallow, Penny royal, Safflower, Peppermint, Hop, Poligi, Pound apple, Marshmallow, Marjoram, Marigold , Malein, Mallow Small, Mallow Blue, Mifuragi, Milk Thistle, Megrinoki, Medu Sweet, Mokka, Yaeyama Aoki, Yakulagi, Yarrow Flower, Eucalyptus, Yakuinin, Mugwort, Raspberry, Lavender, Linden, Le Contains one or more plant extracts selected from Boss, Lemongrass, Lemon Thyme, Lemon Verbena, Lemon Balm, Rosehip, Rose Pink Butts, Rosemary, Rose Red, Laurel, Rafu, Wild Strawberry The present invention provides a glycation inhibitor characterized by the above.

  In addition, the present invention provides a tissue disorder preventive / therapeutic agent containing the glycation inhibitor, a food / beverage product having a tissue disorder preventive / therapeutic effect, and an external preparation.

  The glycation inhibitor of the present invention has a high glycation inhibitory effect.

  Therefore, the glycation inhibitor of the present invention is capable of obtaining prevention, treatment and improvement effects of various tissue disorders caused by glycation or accumulation of terminal glycation substances (AGE) caused by glycation.

FIG. 1 is a drawing showing the body weight of each group of rats per week. FIG. 2 is a drawing showing the daily food intake of rats in each group. FIG. 3 is a drawing showing fluorescence intensity of blood vessels of rats in each group.

  The active ingredients of the glycation inhibitor of the present invention (hereinafter simply referred to as “inhibitor”) are plant extracts shown in Tables 1 and 2 below.

  Among these plant extracts, agrimony, ginkgo, witch hazel, oregano, savory, guava, cumixtin, salacia oblonga, shiso, chanoki, juniper berry, cedar, sage, thyme, clove, sweet leaf sardine, hagoromoso, heather Preferred is an extract of a plant selected from, loquat, elba mate, megsurinoki, eucalyptus, mugwort, raspberry, rabu hemp, rooibos, lemon thyme, lemon balm, rose hip, rosemary, rose red, wild strawberry, guarana, fukugi, In particular, an extract of a plant selected from agrimony, witch hazel, sweet candy coconut, sorghum, sorghum, chanoki, Salacia oblonga, and guarana is preferred.

  These extracts can be used alone or in combination as an active ingredient of the inhibitor of the present invention.

  The method for producing the plant extract is not particularly limited, and can be produced by a commonly used method. Further, the extraction conditions are not particularly limited, and for example, various parts of the plant (whole grass, flowers, buds, seeds, fruits, leaves, branches, bark, root bark, rhizomes, roots, etc.) may be left as they are or cut, ground or After finely pulverized, it is produced by extraction or extraction with a solvent.

  Among these, extraction using a solvent may be performed for about 10 minutes to 1 week under a temperature condition of normal temperature to under pressure and normal temperature to the boiling point of the solvent according to the solvent used. As a solvent used for extraction, a solvent that is usually used in accordance with plant species and treatment steps may be appropriately selected and used. For example, water and alcohols (for example, lower alcohols such as methanol, absolute ethanol, ethanol, etc., or And polyhydric alcohols such as propylene glycol and 1,3-butylene glycol), ketones such as acetone, esters such as diethyl ether, dioxane, acetonitrile, and ethyl acetate, or organic solvents such as xylene, benzene, and chloroform. In addition, when the organic solvent is not preferable to remain, such as when used as a food or cosmetic material, it is particularly preferable to select water or ethanol. These solvents can be used alone, but two or more kinds can be used in any combination.

  Specifically, for example, the following method can be used. That is, after pulverizing the raw material of the plant or the dried product and adding 5 to 20 times the amount of the extraction solvent, it is allowed to stand at normal pressure at room temperature for about one week, or extracted for about 10 to 30 minutes near the boiling point of the extraction solvent. The filtrate obtained by filtration is dried under reduced pressure or freeze-dried to obtain a plant extract.

  The plant extract obtained as described above can be used as it is, but if necessary, purification treatment such as deodorization and decolorization may be further added within a range not affecting the efficacy. Such a purification treatment may be carried out by arbitrarily selecting ordinary means. For example, filtration, ion exchange resin, activated carbon column or the like may be used for adsorption, decolorization, purification and the like. Furthermore, a purified product in the form of a solution, paste, gel, or powder can be obtained by freeze-drying or concentration treatment.

  The inhibitor of the present invention is prepared by formulating the plant extract obtained as described above as it is or in combination with a known pharmaceutical carrier. This inhibitor can be used as a pharmaceutical product such as a tissue disorder preventive / therapeutic agent or a quasi-drug, but it can be used as an external preparation having a tissue disorder preventive / therapeutic effect by blending it into an external preparation such as cosmetics or a food or drink. It can also be a food or drink.

  In order to prepare pharmaceuticals and quasi-drugs, such as tissue damage prevention / treatment agents, which specifically contain the inhibitor of the present invention, the dispersion of the inhibitor of the present invention and ordinary pharmaceuticals and quasi-drugs are used. Mixing with carriers and additives such as adjuvants, excipients, powders, solutions, capsules, suspensions, emulsions, syrups, elixirs, granules, pills, tablets, troches, limonades, etc. It may be in the form of external preparations such as internal preparations, gels, emulsions, creams, ointments, dressings, bathing agents.

  Examples of the carrier include water-soluble monosaccharides such as mannitol, lactose and dextran, oligosaccharides or polysaccharides; gel-forming or water-soluble celluloses such as hydroxypropylcellulose, hydroxypropylmethylcellulose and methylcellulose; crystalline cellulose, α -Water-absorbing and poorly water-soluble celluloses such as cellulose, sodium carboxymethylcellulose and their derivatives; water-absorbing such as hydroxypropyl starch, carboxymethyl starch, cross-linked starch, amylose, amylopectin, pectin and their derivatives And poorly water-soluble polysaccharides; water-absorbing and poorly water-soluble gums such as gum arabic, tragacanth gum, glycomannan and their derivatives; polyvinylpyrrolidone, crosslinked polyacrylic acid and salts thereof, crosslinked polyvinyl alcohol , Polyhydroxyethyl methacrylate and crosslinked vinyl polymers such as derivatives thereof; may be mentioned phospholipids, the lipids sugar to form liposomes, such as molecular aggregate such as cholesterol.

  Additives include microbial culture metabolites, UV absorption / blocking agents, whitening agents, melanin reduction / decomposition agents, turnover promoting effects / cell activators, astringents, active oxygen scavengers, antioxidants, excess Oxidized lipid production inhibitor, anti-inflammatory agent, disinfectant / disinfectant, moisturizer, hair agent, oxidant, dye agent, nutrient enhancer, dairy product, moisturizer, hormones, pH adjuster, chelating agent, antiseptic / Antibiotic agent, refreshing agent, stabilizer, emulsifier, animal / plant protein or its degradation product, animal / plant polysaccharide or its degradation product, animal / plant glycoprotein or its degradation product, blood flow promoter, Antiinflammatory agents / antiallergic agents, cell activators, keratolytic agents, wound treatment agents, foaming agents, thickeners, oral preparations, deodorants / deodorants, bittering agents, seasonings, enzymes, etc. .

  The inhibitor of the present invention has a glycation inhibitory action, and the tissue disorder preventive / therapeutic agent containing the glycation inhibits arteriosclerosis, cataract, Alzheimer's disease caused by glycation, renal failure, dialysis amyloidosis, binding It has the effect of preventing, treating and improving various tissue disorders such as various diseases associated with tissue sclerosis, sunlight elastic fibrosis, and thickening of the skin.

  The dose of the above-mentioned tissue disorder preventive / therapeutic agent may be appropriately adjusted according to the dosage form, disease and the like. 5-60 mass% (hereinafter, simply referred to as “%”), preferably 5-60%, particularly preferably 5-10%. On the other hand, in the case of an external preparation, it may be blended in the range of 0.0001 to 30%, preferably 0.001 to 20%, particularly preferably 0.5 to 20% in terms of solid content in the total amount of the preparation. Furthermore, when using it as a bath agent, the prescription may be taken into consideration so that the concentration is about the same as that of the external preparation for 200 to 300 L of bath water.

  On the other hand, external preparations such as cosmetics containing the inhibitor of the present invention are also produced in the same manner as ordinary external preparations for cosmetics and the like, except that an effective amount of the inhibitor of the present invention is added to cosmetic raw materials. The As the cosmetic raw material to be used, among the above-mentioned agents for preventing or treating tissue damage, those exemplified as carriers and additives that can be used for external preparations such as cosmetics can be used.

  Furthermore, the food / beverage products containing the inhibitor of this invention are manufactured similarly to manufacturing normal food / beverage products except adding the effective amount of the inhibitor of this invention to food-drinks raw materials.

  Examples of these foods and drinks include foods mainly composed of starch such as bread, biscuits, hot cakes, noodles, tablet confectionery, dairy products such as cheese, butter, yogurt, confectionery such as gum, candy, and Japanese confectionery, ham, Examples include meat products such as sausages, fish foods such as chikuwa and kamaboko, seafood products, seasonings such as dressing, soy sauce, jam and sprinkles, beverages such as tea, juice, soft drinks, lactic acid bacteria beverages, alcoholic beverages, etc. The amount of the inhibitor of the present invention in the food or drink is 0.01 to 60%, preferably 0.01 to 20%, particularly preferably about 0.1 to 15% in terms of solid content of the plant extract. It ’s fine.

  EXAMPLES Next, although an Example is given and this invention is demonstrated in detail, this invention is not limited to these at all.

Manufacturing example 1
Production of plant hot water extract:
25 g of each plant described in Table 3 was extracted in 500 mL boiling water under normal pressure for 30 minutes. The obtained extract was filtered with gauze, cooled to room temperature, and lyophilized to obtain a plant hot water extract (Examples 1-111).

Manufacturing example 2
Production of plant ethanol extract:
2.5 g of the dried body of each plant described in Table 4 was extracted in 50 mL of ethanol under normal pressure at room temperature for 1 week. The obtained extract was filtered with gauze, and then the filtrate was dried under reduced pressure to obtain a plant ethanol extract (practical products 112 to 214).

Manufacturing example 3
Production of plant water-containing ethanol extract:
25 g of each plant of witch hazel (stem), rosemary, juniper berry, mugwort, loquat (leaves), and guava (leaves) was extracted in 500 mL of 50% aqueous ethanol at normal pressure for 1 week. The obtained extract was filtered with gauze, and then the filtrate was dried under reduced pressure to obtain a plant-containing ethanol extract.

Test example 1
Measurement of glycation inhibitory effect of plant hot water extract:
The early stage reactant of glycation is a protein-sugar conjugate called an Amadori compound. Furthermore, the initial stage reaction substance forms a terminal saccharification substance (AGE) having fluorescence through dehydration and polymerization. Therefore, the glycation inhibitory action can be measured by measuring the amount of AGE.

  As a specimen, the plant hot water extract (Examples 1-111) obtained in Production Example 1 was used, and the AGE amount was measured by the method of Nakazawa et al. (Nakazawa-H et al., J. Agric. Food Chem. , 48, 180-185 (2000)). In addition, aminoguanidine which is a known glycation inhibitor was used as a positive control in this test.

  Adjust each sample to a final concentration of 10, 2, 0.4, 0.08, 0.016, 0.0032 mg / mL with distilled water, dispense 500 μL each into a test tube with a screw cap, and sterilize by autoclave. And used as a sample. As a positive control, aminoguanidine was prepared to 200, 100, 50, 25, 12.5, 6.25 mM with distilled water, and 500 μL was dispensed into a sterilized test tube while sterilizing the filter. Prepared. Distilled water was used as a control.

  40 mg / mL bovine serum albumin (fraction V; manufactured by Sigma, hereinafter referred to as “BSA”), 1M D-fructose (filtered sterilized in each of the above-mentioned specimen, positive control and control tubes 500 μL of 400 mM potassium phosphate buffer (pH 7.4, final concentration 200 mM) containing 0.5M final concentration was added and reacted at 37 ° C. for 7 days under light shielding. As a blank, a test tube not containing D-fructose was prepared.

  After completion of the reaction, 1 mL of 10% trichloroacetic acid solution was added to each test tube, stirred, cooled and centrifuged (4 ° C., 3000 rpm, 5 minutes), the supernatant was removed by decantation, and a precipitate was further removed. Washed twice with 2 mL of% trichloroacetic acid solution. After washing, the precipitate was dissolved in 2 mL of 200 mM potassium phosphate buffer (pH 7.4). 200 μL of the dissolved precipitate was transferred to a 96-well plate for fluorescence measurement, and fluorescence was measured at an excitation wavelength of 350 nm and a fluorescence wavelength of 425 nm using a fluorescence plate reader SPECTRAmax GEMINI XS (Molecular Device). The rate was calculated.

Furthermore, a logarithmic graph with the horizontal axis representing the sample concentration and the vertical axis representing the glycation inhibition rate was prepared, and the concentration (IC ₅₀ ) showing 50% inhibitory activity was calculated. The obtained results are shown in Table 3.

(Result)
As shown in Table 3, each plant extract inhibited the formation of AGE and an effect of inhibiting glycation was observed. In particular, witch hazel (stem), juniper berry, sorghum, rooibos, rose red, witch hazel (leaves), canola (stems and leaves), savory, guava (leaves), heath, candy leaf coconut, eucalyptus, clove, agrimony, Oregano, raspberry (leaves), wild strawberry, lemon balm, rosemary, thyme, mugwort, salacia oblonga, sage, ginkgo (leaves), cedar (leaves), lemon thyme (leaves), megsurinoki, cumixtine, loquat (leaves), The IC ₅₀ of Elba Mate, Shiso, Mannenrou, Rosehip and Rafu is equivalent to or higher than the IC _{50 of} aminoguanidine, 0.28 mg / mL, and has an excellent glycation-inhibiting action. confirmed.

Test example 2
Measurement of glycation inhibitory action of plant ethanol extract (1):
As the specimen, the plant ethanol extract (practical products 112 to 214) obtained in Production Example 2 was used. The measurement was performed by partially modifying the method of Test Example 1. That is, each specimen was adjusted to 0.4 mg / mL with ethanol, and 500 μL was dispensed into a sterilized test tube while sterilizing the filter, thereby preparing a sample. Further, as a positive control, aminoguanidine was adjusted to 0.4 mg / mL with distilled water, and 500 μL was dispensed into a sterilized test tube while sterilizing with a filter.

  On the other hand, BSA was dissolved in 0.4 M potassium phosphate buffer (pH 7.4) so as to be 40 mg / mL and D-fructose to be 1 M, and further 4 volumes of distilled water were added to 5 volumes of this solution. Then, it sterilized with the filter and was set as the BSA-fructose reaction liquid.

  In the test, 900 μL of the BSA-fructose reaction solution was aseptically dispensed into an autoclave-sterilized screw-mouth test tube, and 100 μL of a sample or a positive control was further added. These test tubes were reacted at 37 ° C. for 7 days under light shielding. In addition, the thing which added only 100 microliters of ethanol was used for the control | contrast, and the BSA solution which does not contain D-fructose was prepared for the blank.

  After completion of the reaction, 1 mL of 10% trichloroacetic acid solution was added to each test tube, stirred, cooled and centrifuged (4 ° C., 3000 rpm, 5 minutes), the supernatant was removed by decantation, and a precipitate was further removed. Washed twice with 2 mL of% trichloroacetic acid solution. After washing, the precipitate was dissolved in 2 mL of 200 mM potassium phosphate buffer (pH 7.4). 200 μL of this solution was transferred to a 96-well plate for fluorescence measurement, and fluorescence was measured at an excitation wavelength of 350 nm and a fluorescence wavelength of 425 nm using a fluorescence plate reader SPECTRAmax GEMINI XS (Molecular Device). (Maillard reaction inhibition rate) was calculated. Table 4 shows the obtained results.

(Result)
As shown in Table 4, even when ethanol was used as the extraction solvent, each plant extract inhibited the formation of AGE and an effect of inhibiting glycation was observed. In particular, witch hazel (stem), witch hazel (leaves), coral spruce, fukugi, guarana, sweet leaf scallop, tea tree (stems and leaves), agrimony, salacia oblonga, and megsurinoki are equivalent to or higher than aminoguanidine. It was confirmed that it has an excellent glycation inhibitory action.

Test example 3
Measurement of glycation inhibitory action of plant ethanol extract (2):
Among the samples obtained in Production Example 2, the witch hazel (stem), witch hazel (leaves), caterpillar, fukugi, guarana, sweet candy coconut, chanoki (stems and leaves) that were highly effective in Test Example 2 were obtained. ), Agrimony and Salacia Oblonga. These specimens were adjusted to 0.2 to 0.002 mg / mL with ethanol and subjected to the test. According to the method of Test Example 2, the glycation inhibition rate and IC ₅₀ were calculated. The results obtained are shown in Table 5.

(Result)
As shown in Table 5, any plant extract inhibited the formation of AGE and showed an excellent glycation-inhibiting effect. In particular, witch hazel (stem), witch hazel (leaves), coral spruce, fukugi, guarana, sweet potato coconut, chanoki (stems and leaves), agrimony is equivalent to or higher than aminoguanidine and has a low concentration. Was confirmed to have an excellent glycation-inhibiting action.

Manufacturing example 4
Preparation of witch hazel (stem) hot water extract:
50 g of witch hazel (chopped dry bark) was weighed and placed in 1 L of boiling water and boiled for 30 minutes. The mixture was allowed to cool at room temperature, and then poured with three layers of gauze, and the extract was freeze-dried to obtain 6.8 g of extract powder (yield 13.6%).

Manufacturing example 5
Preparation of witch hazel (leaf) hot water extract:
50 g of witch hazel (dried dried leaves) was weighed and placed in 1 L of boiling water and boiled for 30 minutes. The mixture was allowed to cool at room temperature, and then poured with three layers of gauze, and the extract was freeze-dried to obtain 7.2 g (yield 14.4%) of an extract powder.

Manufacturing Example 6
Preparation of hot water extract:
Weighed 50 g of sorghum (chopped and dried whole plant), placed in 1 L of boiling water and boiled for 30 minutes. The mixture was allowed to cool to room temperature, and then poured with three layers of gauze, and the extract was lyophilized to obtain 10.6 g of extracted powder (yield 21.2%).

Manufacturing example 7
Preparation of witch hazel (stem) ethanol extract:
2.5 g of witch hazel (chopped dry bark) was weighed, immersed in 50 mL of ethanol, and allowed to stand at room temperature for 1 week under light shielding. After rinsing with filter paper, the filtrate was placed in a 100 mL eggplant flask and dried under reduced pressure with a rotary evaporator to obtain 230 mg of ethanol extract (9.2% yield).

Manufacturing Example 8
Preparation of witch hazel (leaf) ethanol extract:
2.5 g of witch hazel (dried dried leaves) was weighed, immersed in 50 mL of ethanol, and allowed to stand at room temperature for 1 week under light shielding. After rinsing with filter paper, the filtrate was put into a 100 mL eggplant flask and dried under reduced pressure with a rotary evaporator to obtain 360 mg of ethanol extract (yield 14.4%).

Manufacturing example 9
Preparation of sorghum ethanol extract:
We measured 2.5 g of sorghum (cut chopped whole plant), dipped in 50 mL of ethanol, and allowed to stand at room temperature for 1 week in the dark. After dripping with filter paper, the filtrate was put into a 100 mL eggplant flask and dried under reduced pressure with a rotary evaporator to obtain 160 mg (yield 6.4%) of an ethanol extract.

Manufacturing example 10
Preparation of raspberry (fruit) hot water extract (1):
To 60 g of raspberry (raw fruit), 200 mL of ion-exchanged water was added and boiled under normal pressure for 30 minutes to obtain an extract. The extract was then cooled to room temperature, filtered through filter paper, and lyophilized to obtain 3.14 g of hot water extract (yield 5.2%).

The hot water extract obtained above was measured for glycation inhibitory action in the same manner as in Test Example 1. As a result, the IC ₅₀ of the raspberry (fruit) hot water extract was 0.112 mg / ml.

Manufacturing Example 11
Preparation of raspberry (fruit) hot water extract (2):
200 mL of ion-exchanged water was added to 57.8 g of raspberries (frozen fruits), and the mixture was boiled for 30 minutes under normal pressure to obtain an extract. The extract was then cooled to room temperature, filtered through filter paper, and lyophilized to obtain 4.8 g of hot water extract (yield 8.3%).

The hot water extract obtained above was measured for glycation inhibitory action in the same manner as in Test Example 1. As a result, the IC ₅₀ of the raspberry (fruit) hot water extract was 0.108 mg / ml.

Test example 4
Measurement of glycation inhibitory effect of plant hot water extract in animal body:
As specimens, the guava (leaf) hot water extract (practical product 9) and the wild strawberry (leaf) hot water extract (practical product 17) obtained in Production Example 1 were used. As animals, 18 6-week-old male stroke-prone hypertensive rats (SHR-SP / Izm) were used. The rats were divided into 3 groups (6 animals each) administered with control, guava (leaves) hot water extract (GvEx), and wild strawberry (leaves) hot water extract (WSEx) after 1 week of preliminary breeding. The feed having the composition shown in Table 6 was fed for 4 weeks and reared. The weekly body weight and daily food intake of each group of rats were measured. The results are shown in FIG. 1 and FIG.

  After anesthetizing each group of rats after 4 weeks of breeding, they were exsanguinated and the thoracic aorta was removed. The extracted aorta was degreased, freeze-dried, and finely crushed. Next, an aliquot of this was taken and treated with pronase for 24 hours, and then the supernatant was recovered and used as a sample. The fluorescence intensity of this sample was measured at an excitation wavelength of 370 nm and an emission wavelength of 430 nm, and the amount of AGE was calculated. The results are shown in FIG. The relationship between the fluorescence intensity and the AGE amount is described, for example, in the literature (J. Hypartension, 17 (4), 1999., p483, left line 8-24).

(Result)
There was no significant difference between the weekly body weight and the daily food intake of each group of rats. Moreover, the amount of AGE in the blood vessels was decreased in both the WSEX group and the GvEx group as compared with the control group.

  Hereinafter, formulation examples of the glycation inhibitor according to the present invention will be described as typical examples.

Example 1
Latex:
An emulsion was prepared using the hydrated ethanol extract of plants of witch hazel (stem), rosemary, juniper berry and mugwort obtained in Production Example 3.

(Component) (mass%)
1. Squalane 5.0
2. Olive oil 5.0
3. Jojoba oil 5.0
4). Cetyl alcohol 1.5
5). Glycerol monostearate 2.0
6). Polyoxyethylene (20) cetyl ether 3.0
7). Polyoxyethylene (20) soorbitan monooleate 2.0
8.1,3-Butylene glycol 1.0
9. Glycerin 2.0
10. Plant water-containing ethanol extract 5.0
11. Perfume appropriate amount 12. Preservative appropriate amount13. Purified water balance

  The obtained emulsion was extremely useful with excellent usability.

Example 2
Granule bath preparation:
A granule bath preparation was prepared using plant ethanol extracts of witch hazel (stem), rosemary and mugwort plants obtained in Production Example 2.

(Component) (mass%)
1. Sodium bicarbonate 60.0
2. Anhydrous sodium sulfate 30.0
3. Borax 5.0
4). Plant ethanol extract 5.0

  The obtained bath agent was extremely useful with excellent usability.

Example 3
Drug:
The following drugs were mixed and the mixture was tableted. In addition, the implementation product which confirmed the glycation inhibitory effect in Test Example 3 was used as a plant extract.

(Components) (Mass per tablet (mg))
1. Plant extract 10
2. Lactose 190
3. Potato starch 39
4). Microcrystalline cellulose 30
5). Synthetic aluminum silicate 30
6). Calcium stearate 1

  The obtained tablet was stable and easy to take.

Example 4
Soft drinks:
100 mg of hot water extract of Hagoromoso obtained in Production Example 6 and 5 g of sugar were mixed with 100 mL of sodium carbonate water to produce a soft drink.

  The obtained soft drink was a light yellow drink and had a refreshing taste.

Example 5
chocolate:
100 mg of milk extract obtained in Production Example 5 was added to 100 g of milk chocolate melted in a hot water bath, stirred well, poured into a mold, cooled and hardened.

  The obtained chocolate had a good texture and good taste, and was inferior to ordinary chocolate.

  The glycation inhibitor of the present invention has a high glycation-inhibiting effect, and has the effect of preventing, treating, and improving various tissue disorders caused by glycation or accumulation of terminal glycation substances (AGE) caused by glycation. It is something that can be done. Moreover, since the active ingredient of the glycation inhibitor of the present invention is derived from a plant, it is highly safe.

Therefore, the glycation inhibitor of the present invention can be used as a pharmaceutical product or a quasi-drug, and can be blended in cosmetics, foods and drinks, and the like.
more than

Claims (7)

  Agrimony, Ashwagandha, Anise, Achacharu, Gancho Sugo, Ginkgo, Fennel, Witch Hazel, Turmeric, Echinacea, Ezocoggi, Elder, Elba Mate, Olive, Oregano, Orange, Moth, Oyster, Chamomile, Galana , Gymnema, Cat's Claw, Catnip, Killaway, Kingfisher, Guava, Cuttlefish, Beardness, Cumixtin, Black Soybeans, Hoes, Cinnamon, Ketchmeishi, Goya, Cola Nuts, Gokahi, Gotsukora, Coriander, Comfrey, Saximasow, Saflower, Salacia・ Oblonga, hawthorn, sanitary carrot, perilla, juyaku, juniper berry, ginger, cedar, horsetail, stevia, spearmint, sage, savory, hawthorn, sensation, thyme, tahibo Dandelion, Chervil, Chanoki, Chabotodia, Clove, Daisy, Dill, Eucommia, Nikuzuku, Nanto, Nettle, Egret, White Flower Serpentine Grass, Lagoon, Basil, Lotus, Half Branch, Heath, Peach, Hyssop, Loquat, Feverfew, Fukugi, Wisteria, Plantago Thyrium, Blueberry, Blue Mallow, Penny Royal, Safflower, Peppermint, Hop, Polyge, Pound Apple, Marshmallow, Marjoram, Marigold, Malene, Mallow Small, Mallow Blue, Miffuragi, Milk Thistle , Medu Sweet, Mokka, Yaeyama Aoki, Yakurakugi, Yarrow Flower, Eucalyptus, Yokuinin, Artemisia, Raspberry, Lavender, Linden, Rooibos, Lemongrass, Lemon Thyme, Lemon Verbena, Remo An extract extracted from a plant selected from balm, rosehip, rose pink butts, rosemary, rose red, laurel, rabu hemp, and wild strawberry, using an organic solvent or a combination of organic solvent and water. A glycation inhibitor containing one or more kinds.   The glycation inhibitor according to claim 1, wherein the plant is selected from agrimony, witch hazel, candy leaf coconut, sorghum, chanoki, Salacia oblonga, and guarana.   The glycation inhibitor according to claim 1, wherein the organic solvent is ethanol.   The glycation inhibitor according to any one of claims 1 to 3, which is an additive for food and drink.   A preventive or therapeutic agent for tissue damage comprising the glycation inhibitor according to claim 1.   A food / beverage product having a tissue disorder preventing / treating effect comprising the glycation inhibitor according to claim 1.   An external preparation containing a glycation inhibitor according to claim 1 and having an effect of preventing and treating tissue damage.

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