JP2015160842A - Saccharification reaction inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a saccharification reaction inhibitor that has excellent saccharification reaction inhibitory action, and particularly inhibits the reaction among an intermediate, and an amino acid, a peptide, a protein or salt thereof in the saccharification reaction.SOLUTION: A saccharification reaction inhibitor comprises one or more plant selected from the group consisting of Japanese butterbur, chestnut, peanut, Persimmon, rose hip, blueberry, pomegranate, rosemary, grape, apricot, perilla, sage, peppermint, savory, Yerba mate, coffee tree, olive, and Camellia sinensis or a processed product thereof.

Description

  The present invention relates to a saccharification reaction inhibitor.

  Proteins are responsible for various physiological functions in vivo. However, in vivo, there is a reaction called Maillard reaction or saccharification reaction in which proteins and amino acids in the tissue are saccharified, and in some cases, it may be accumulated in the body as an abnormal protein. For example, if a metabolic function decline due to aging or a hyperglycemic state due to the onset of diabetes continues, the saccharification reaction gradually proceeds in vivo, and the normal function of the protein in each tissue is impaired. When a saccharification reaction occurs, an irreversible compound called an advanced glycation end-products (AGE) is finally formed, and is deposited in each tissue or is localized in vascular endothelial cells. It is known to bind to the body and cause various diseases.

  Diseases that are thought to contribute to protein glycation include diabetic complications, nephropathy, retinopathy, neuropathy, Alzheimer's disease, arteriosclerosis, malignant tumor, bone disease, neurodegenerative disease, etc. Yes. Skin aging is also thought to be due to protein glycation.

  Furthermore, in food and drink containing protein and sugar, for example, there is a problem that browning or precipitation of the food or drink is caused by a saccharification reaction during storage. Therefore, inhibiting glycation of proteins is thought to be effective in preventing and treating these diseases and symptoms and preventing degeneration of food and drink during storage. Various studies have been conducted so far. Various active ingredients are known as saccharification reaction inhibitors (see Patent Documents 1 to 3).

JP 2013-213021 A JP 2013-23487 A JP 2009-84214 A

  As the reaction pathway of the saccharification reaction, methylglyoxal (MGO), 3-deoxyglucosone (3-deoxyglucosone (3-GO) produced by the reaction of a reducing sugar such as glucose with a protein or amino acid, and dehydration or hydrolysis of the produced Amadori compound. There is known a pathway in which an intermediate such as DG) reacts with a protein or an amino acid to produce AGE through a dehydration / polymerization reaction. Also known is a pathway in which a carbonyl compound such as glyoxal (GO) produced by an automatic oxide or decomposition product of glucose reacts with a protein or an amino acid to produce AGE through a dehydration / polymerization reaction. As AGEs, glucose-derived AGE, glyceraldehyde-derived AGE, glycolaldehyde-derived AGE, MGO-derived AGE, GO-derived AGE, and 3-DG-derived AGE are known.

  In many studies conducted so far to search for substances that inhibit the saccharification reaction described in Patent Documents 1 to 3, the saccharification end product (AGE) is focused on, and the saccharification end product (AGE) is generated. The inhibitory effect of the saccharification reaction is determined by the amount. However, little research has been conducted to search for substances that inhibit the saccharification reaction by focusing on the reaction pathway of the intermediate of the saccharification reaction.

  An object of the present invention is to provide a saccharification reaction inhibitor that inhibits a saccharification reaction by inhibiting a reaction between an intermediate in the saccharification reaction and a protein or amino acid.

  As a result of intensive studies by the present inventors to solve the above-mentioned problems, a specific plant extract is reacted with amino acid, peptide, protein or a salt thereof in methylation reaction and methylglyoxal (MGO). It discovered that reaction which produces | generates a saccharification reaction product was inhibited, and came to complete this invention.

  The present invention is selected from the group consisting of Japanese cypress, chestnut, peanut, oyster, rosehip, blueberry, pomegranate, rosemary, grape, apricot, perilla, sage, peppermint, savory, yerba mate, coffee, olive, chanoki Provided is a saccharification reaction inhibitor containing at least one plant or processed product thereof.

  The said saccharification reaction inhibitor has the effect | action which inhibits reaction with the intermediate in a saccharification reaction, and an amino acid, a peptide, protein, or these salts. Examples of the intermediate include methylglyoxal.

  ADVANTAGE OF THE INVENTION According to this invention, the saccharification reaction inhibitor which has the outstanding saccharification reaction inhibitory effect can be provided.

[Saccharification reaction inhibitor]
In the present invention, saccharification reaction inhibition means that an Amadori compound produced by a reaction of a saccharide with an amino acid, a peptide or protein containing this as a constituent component and a salt thereof (hereinafter also referred to as “amino acids”) is oxidized, dehydrated, It means to inhibit the reaction to produce AGE through a reaction such as condensation. In the present invention, attention is focused on saccharification reaction inhibition that inhibits the production of AGE by inhibiting the reaction between an intermediate in the saccharification reaction and amino acids. The following reaction is mentioned as reaction which produces | generates AGE by reaction with the intermediate body and amino acids in a saccharification reaction. i) Intermediates such as methylglyoxal (MGO) and 3-deoxyglucosone (3-DG) produced by dehydration and hydrolysis of the Amadori compound produced by the reaction of saccharides with amino acids are amino acids Ii) a reaction that produces AGE through dehydration and polymerization reactions, and ii) a carbonyl compound such as glyoxal (GO) produced by an auto-oxide or decomposition product of glucose, which is an amino acid Reaction that produces AGE through dehydration and polymerization reaction. Here, the saccharide means a reducing sugar, and examples thereof include monosaccharides such as glucose, fructose, xylose and arabinose, and disaccharides such as maltose and lactose.

  In particular, the present invention provides a saccharification reaction inhibitor having an excellent effect of inhibiting the reaction between methylglyoxal (MGO), which is an intermediate, and amino acids.

(Plant or processed product)
The saccharification reaction inhibitor of the present invention consists of cypress, chestnut, peanut, oyster, rosehip, blueberry, pomegranate, rosemary, grape, apricot, perilla, sage, peppermint, savory, yerba mate, coffee, olive, chanoki It contains at least one plant selected from the group or a processed product thereof as an active ingredient.

  As used in the present invention, “Fuki” is a plant belonging to the genus Petasites, and wild species such as Petasites japonicus, cultivars such as mizubuki, aichibuki, akabuki can be used. In the present invention, the site of the balloon used as a raw material is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor. In addition, the above-ground part of a balloon may be a flower stem (Fukinotou), a fruit, etc. As the strain type, either a male strain or a female strain may be used. In addition, the flowers appear in the early spring prior to the development of the leaves, and a flower head (flower stem: Fukinoto) is attached to the top of a short upright stem in a bunched form. In the present invention, Fukinoto peel is preferably used.

  The “chestnut” used in the present invention is a plant belonging to the family Beechaceae (Fagaceae) or Chestnut genus (Castanea), such as chestnut (also known as Japanese chestnut) (Castanea crenata Sieb. Et Zucc.), American chestnut (Castanea dentata ( Marsh.) Borkh = (C. Americana Raf.)), Castanea mollissima Bl.nea, etc. can be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but a seed coat (endothelium or outer coat) or the like is preferably used.

  The “peanuts” used in the present invention are seeds of arachis hypogaea belonging to the order of the order of roses, legumes, and peanuts, and any known peanuts can be used. As the types of peanuts, for example, Tachi 1, improved Wadaoka, Tachi camel 1, Chiba Hanate, Chiba 55, Chiba 43, Chiba 74, Java 13, white oil 7-3, full grain, Valencia, 334A, Azuma handkerchief, Tecona, Waseda ryu, Benihandachi, Sachihomare, Tachimasari, etc. are known. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but seed coat (thin skin) or the like is preferably used.

  The “oyster” used in the present invention is a plant belonging to the family Ebenaceae and Diospyros, and other species of the same genus (Diospyros kaki L.) and other species of the same genus (D. discolor Willd.) (Diospyros eriantha Champ. Ex Benth.), Kuroki (D. ferrea (Willd.) Bakh. (= Maba buxifolia Pers.)), Shinoaki (D. lotus L.), Diospyros morrisiana Hance, D. atropurpurea Guerke, D.D. conzattii Standl. , D. ebenum Koenig, D.D. virginiana L. Etc. can also be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but seeds, straw, etc. are preferably used.

  The “rose hip” used in the present invention is a plant of the Rosaceae family or the genus Rosa. Rose hip (also known as: rose, dog rose, dog rose, dog briar, canina rose) (Rosa canina L.). ), Egrantin rose (aka Sweetbrier, Egrantine) (R. eglanteria (= R. Rubiginosa)), Rosamicranta (R. micrantha Borrer ex Sm.), Hamanashi (aka: Japanese rose, Turkey turn) Rose (R. rugosa Thunb.), Teri Hanoi rose (R. wichuraiana Crep.), Neubara (aka: Ages) (R. multiflora Thunb.), Yamaha Manas (aka: Calaftobara) (R. davurica Pallas (= R. Willdenowii) Sprengel)), Naniwaibara (R. laevigata Michx.), R. bella Rehd. Et Wils. , R. sweginzowii Koehne, Kogane Hide (R. cymosa Tratt.), Kim Saeko, R. odorata Sweet var. gigantea Rehd. Et Wils. macrophylla Lindl. R. maikai (= R. rugosa var. Plena Regel), Rika ria (R. banksiae Ait. Var. Normalis Reg.), Kakayanbara (R. bracteata Wendl.), Koushinbara (R. chinensis Jacq. ), Small infarction rose (R. graciliflora Rehd. Et Wils.), Purple rose (R. multiflora Thunb. Var. Platyphylla Thory), R. omeiensis Rolfe, Izayoi rose (also known as stamen) (R. f. normalis Rehd. et Wils), R. sericea Lindl., R. sertata Rolfe, R. hybrida Hort., R. centifolia L. ), Damascena Miller, R. damascena Miller, R. moschata Herr., R. banksiae Aiton, R. foetida Herr. (= R. Lutea Mill.) )), Gigantea rose (R. gigantea) Collett et Hemsl.), R. odorata Sweet, R. acicularis Lindl, etc. can be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but whole plants and the like are preferably used.

  The “blueberry” used in the present invention is a plant belonging to the family Aceraceae (Ericaceae), Vaccinium, Lowbush blueberry (Vaccinium angustifolium Ait.), And other species of the same species, Rabbit eye blueberry (Vaccinium ashei Reade.), High bush blueberry (Vaccinium australe Small.), Bilberry (Vaccinum myrtillus L.), blackberry (Vaccinum uliginosum L.) and the like can be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but leaves and the like are preferably used.

  The `` pomegranate '' used in the present invention is a plant of the family Pomegranate (Punicaceae), Pomegranate (Punica), pomegranate (Punica granatum L.), and other species belonging to the same species, P. granatum L. var. Pers.) Etc. can also be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but pericarp or the like is preferably used.

  The “rosemary” used in the present invention is a plant belonging to the family Labiatae and genus Rosmarinus, and rosemary (Rosmarinus officinalis L.) or the like can be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but a branch or the like is preferably used.

  The `` grape '' that can be used in the present invention is a plant of the genus Vitaceae Vitis, grape (Vitis vinifera L.), American grape (Vitis labrusca L.), Amazuru (V. saccharifera Makino) ), Shrimp (V.ficifolia Bunge var.lobata (Regel) Nakai), sanctuary (V.flexuosa Thunb.), Wild grape (V.coiguetiae Pulliat), European and American hybrid grapes (V. it can. Grape varieties are not limited, and for example, Delaware, Campbell, Pione, Berry A, Kyoho etc. can be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but branches, fruits, fruit peels, seeds and the like are preferably used.

  `` Apricot '' used in the present invention is a plant of the Rosaceae family (Rosaceae), the genus Sakura (Prunus), apricot (Prunus armeniaca Linne var. Anzu Maximowicz), and other species of the same genus Honzu (P. armeniaca Linne) , American red plum (P. americana Marsh.), Atlantic cherry (P. avium L.), Sumi cherry (P. cerasus L.), Plum (P. domestica Lindl.), Plum (P. salicina Lindley), Santou (P.persica Batsch var.davidiana Maximowicz), common peach (P.persica Batsch var.persica), nectarine (P.persica Sieb.et Zucc.var.nucipersica Schneid.), Etc. can be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but leaves, seed coats and the like are preferably used.

  “Perilla” used in the present invention is a plant belonging to the family Labiatae, Perilla, Perilla frutescens Britton var. Acuta Kudo, and its modified species, P. frutescens Britton var. crispa Decne.), and related plants such as Aojiso (P. frutescens viridis) and Katamejiso (P. frutescens f. discolor). The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but leaves and the like are preferably used. In addition, Aojiso includes what is called Ooba in Japan.

  The “sage” used in the present invention is a plant of Labiatae, and can be used for Salvia sclarea L., sage (Salvia officinalis) and the like. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but whole plants and the like are preferably used.

  “Peppermint (also known as peppermint pepper, pepper pepper)” used in the present invention is a plant belonging to the family Lamiaceae or Mentha, and peppermint (Mentha piperita L.) or the like can be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but whole plants and the like are preferably used.

  The “savory” used in the present invention is a plant belonging to the genus Satureja, and savory (Satureja hortensis L.) and the like can be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but whole plants and the like are preferably used.

  The “Yerba mate” used in the present invention is a plant belonging to the genus Ilex paraquariensis, and Yelba mate (Ilex paraquariensis) and the like can be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor. Leaves and stems are preferred because they are easily available as mate tea.

  The “coffee tree” used in the present invention is a plant belonging to the family Rubiaceae and Coffee genus (Coffea). liberica) can be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor. The coffee beans that are the seeds of the coffee tree are preferably roasted or used as they are.

  The “olive” used in the present invention is a plant belonging to the family Oleaceae, such as olive (Olea europaea L.), wild-type olive (O. europaea var. Sylvestris (Mill.) Brotero), and the genus Olea chrysophylla Lam. Etc. can be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but fruits, leaves and the like are preferably used.

  As used in the present invention, "Chinoki" is a plant of the family Camellia (Theaceae), the genus Chanoki (Thea), and is the same assamcha (Thea sinensis L. (= Camellia sinensis L.)) and other species of the same genus ( T. sinensis var. Assamica (= Camellia sinensis var. Assamica), toucha (T. macrophylla (= Camellia sinensis var. Sinensis f. Macrophylla)), etc. can be used. The site to be used is not limited as long as it is useful as an active ingredient of a saccharification reaction inhibitor, but leaves and the like are preferably used. In addition, Yame tea is preferably used as the tea to be used.

  The said plant or its processed material can be used as an active ingredient of the saccharification reaction inhibitor of this invention by 1 type (s) or 2 or more types. In addition, these plants may use waste at the time of production by thinning or fruit picking, or other waste generated at the time of processing.

  In the present invention, the plant or processed product thereof refers to each part of the above-mentioned raw materials (whole plant, flower, flower stem, seed, seed coat, fruit, pericarp, persimmon, stem, bud, leaf, branch, branch leaf, trunk, bark, sap, Root bark, rhizomes, roots, or a part of these parts) are usually dried and then pulverized, extracted, purified, concentrated, and dried (depending on the form and intended dosage form). It means a product prepared as a processed product after being subjected to various processing treatments such as spray-drying treatment and freeze-drying treatment.

  Examples of the processed product in the present invention include, for example, a pulverized processed product (which may be either coarse powder or fine powder), an extract extracted with various solvents, a dried product (dried extract), and a powder. And dry powder extract.

  When the processed product in the present invention is a pulverized processed product, the manufacturing method thereof is not limited and can be manufactured by a conventionally known method or the like. For example, each part of the plant can be dried by constant temperature drying (drying using a constant temperature device, etc.), hot air drying, freeze drying, etc., and the resulting dried product can be subjected to a pulverizer or the like to obtain a pulverized processed product. .

  When the processed product in the present invention is an extract, its production method is not limited and can be produced by a conventionally known method or the like. For example, an extract can be obtained by extracting or solvent extraction after each part of the plant as it is or after cutting, pulverizing, or the like. As a method for solvent extraction, a method known in the art may be adopted, and for example, a conventionally known extraction method such as water extraction, hot water extraction, hot water extraction, alcohol extraction, supercritical extraction or the like may be used. it can.

  When performing solvent extraction, the solvent is, for example, water; lower alcohols such as methanol, anhydrous ethanol, ethanol, and alcohols such as polyhydric alcohols such as propylene glycol and 1,3-butylene glycol (whether anhydrous or hydrous) Ketones such as acetone, esters such as diethyl ether, dioxane, acetonitrile, and acetic acid ethyl ester, xylene, benzene, chloroform, and the like are preferable, and water, ethanol, and the like are preferable. These solvents may be used alone or in combination of two or more.

  The obtained extract may be used as it is or may be dried and powdered. Moreover, you may refine | purify, a concentration process, etc. to the extract obtained as needed. As the purification treatment, treatment such as filtration or adsorption or decolorization using an ion exchange resin or activated carbon column can be performed. As the concentration treatment, a conventional method such as an evaporator can be used.

  Alternatively, the obtained extract (or purified product or concentrate) is subjected to lyophilization and powdered, or excipients such as dextrin, corn starch, gum arabic and the like are added and powdered by spray drying. It may be pulverized according to a conventionally known method such as a method to obtain a processed product used in the present invention. Moreover, you may use this processed material by melt | dissolving in a pure water, ethanol, etc. as needed.

  As a plant-derived processed product used in the present invention, a commercially available product may be used simply.

  The blending amount of the processed product in the saccharification reaction inhibitor of the present invention is not particularly limited as long as the effect of the present invention is exhibited. For example, 0.0001 to 100% by weight in terms of the dry weight of the processed product, preferably 0.00. It is 001-50 weight%, More preferably, it is 0.01-20 weight%.

(Other ingredients)
In the saccharification reaction inhibitor of the present invention, the above-mentioned active ingredient can be used alone, but besides the above-mentioned ingredients, conventionally known fats and oils, waxes, hydrocarbons, fatty acids, alcohols, esters, interfaces Components such as an activator, a metal soap, a pH adjuster, an antiseptic, a fragrance, a powder, an ultraviolet absorber, a thickener, a dye, an antioxidant, a saccharification reaction inhibitor, and a chelating agent can be blended.

(Use)
The saccharification reaction inhibitor of the present invention can be used, for example, in a composition for oral administration or ingestion or a composition for external use on the skin. The compounding amount of the saccharification reaction inhibitor according to the present invention depends on the type of product to be formulated, the age, sex, body weight, condition, administration time, dosage form, method of administration or ingestion, timing of administration or ingestion, etc. It can be set appropriately. The composition for oral administration or ingestion, and the content of a plant or processed product thereof as an active ingredient of a saccharification reaction inhibitor in a composition for external use on skin is 0.0001 to 100% by weight in terms of dry weight of the processed product, Preferably it is 0.001 to 50 weight%, More preferably, it is 0.01 to 20 weight%.

  Compositions for oral administration or ingestion are shaped into capsules such as hard capsules and soft capsules, tablets and pills, or powders, granules, and bowls according to consumer preferences Can be done. It can also be prepared in liquid dosage forms or forms such as solutions, suspensions, or emulsions. The composition for oral administration or ingestion can be used as a pharmaceutical, a quasi-drug, a food for specified health use, a dietary supplement, other food or drink, or can be used in combination with these. The composition for oral administration or ingestion may be eaten as it is or may be taken by dissolving in water, hot water, milk, soy milk, tea, juice or the like according to the dosage form or shape or preference.

  By taking a composition containing the saccharification reaction inhibitor of the present invention on a daily basis, prevention or improvement of diseases or symptoms caused by glycation of amino acids in vivo can be expected.

  When the composition containing the saccharification reaction inhibitor of the present invention contains an amino acid, peptide, protein or salt thereof, the saccharification reaction of the amino acids is suppressed by including the saccharification reaction inhibitor, and the food or drink , Browning, precipitation, etc. can be suppressed, and quality stability can be improved.

  The external composition for skin can be processed into various forms such as lotions, emulsions, gels, creams, ointments and the like. Skin external preparation compositions are used as cosmetics, pharmaceuticals, quasi drugs, and the like. Specifically, the saccharification reaction inhibitor used in the present invention includes lotion, cosmetic cream, milky lotion, cream, pack, hair tonic, hair cream, shampoo, hair rinse, treatment, body shampoo, facial cleanser, soap, foundation, It can be used by being incorporated into a hair restorer, aqueous ointment, or spray.

  Since the saccharification reaction inhibitor of the present invention has an excellent saccharification reaction inhibitory effect, the composition for external use containing the skin inhibits the saccharification of amino acids in the skin and causes skin wrinkles, spots, sagging, etc. Aging can be prevented. In addition, by preventing skin aging, the effect of improving skin gloss and firmness can be expected.

  Hereinafter, although a test example is shown and this invention is demonstrated in detail, this invention is not limited to these.

[Preparation of plant-derived processed products]
A plant extract was prepared by water extraction, hot water extraction, or ethanol extraction using a plant-derived processed product shown in Table 1 below after freeze-drying the plant material and using the pulverized plant material as an extraction plant material. Of the plants listed in Table 1, Aojiso, Rosemary, Pomegranate, Apricot, Grapes (Kyoho, Delaware, Pione, Berry A, Campbell), Oyster, Blueberry, Chestnut, Chanoki, Olive (leaves) The thing of the prefecture was used.

  In water extraction, 1 g of plant raw material for extraction is added to 20 mL of ultrapure water, subjected to ultrasonic waves for 5 minutes, and then stirred overnight at a temperature of 4 ° C., followed by centrifugation (rotation speed: 3000 rpm, 20 minutes) ). Thereafter, the supernatant was collected. Moreover, about the deposit, this was added to 10 mL of ultrapure water, it centrifuged again (rotation number 3000rpm, 20 minutes), and the supernatant was added to the supernatant collect | recovered by the previous centrifugation. The supernatant collected in this manner was freeze-dried and then frozen and stored as a plant extract.

  In the hot water extraction, the step of stirring in the water extraction at a temperature of 4 ° C. overnight and then centrifuging is performed after stirring for 1 hour in a state of maintaining the temperature at 95 ° C. Except for the point of changing to the step of 3000 rpm for 20 minutes, the same procedure as in the water extraction was performed, and the collected supernatant was freeze-dried and then frozen and stored as a plant extract.

  In ethanol extraction, 1 g of plant material for extraction was added to 20 mL of ethanol, subjected to ultrasonic waves for 5 minutes, stirred at room temperature for 1 hour, and then centrifuged (rotation speed: 3000 rpm, 5 minutes). Thereafter, the supernatant was collected. Moreover, about the deposit, this was added to 10 mL of ultrapure water, it centrifuged again (rotation speed 3000rpm, 5 minutes), and the supernatant was added to the supernatant collect | recovered by the previous centrifugation. A plant extract was obtained by freezing and storing the collected supernatant.

[Calculation method for inhibition rate of saccharification reaction]
As one route of the saccharification reaction, there is a reaction in which methylglyoxal (MGO), which is an intermediate, reacts with a protein to form an MGO-protein conjugate. Thereafter, such an MGO-protein conjugate undergoes oxidation, dehydration, condensation and the like to form a saccharification end product (AGE). Some MGO-protein reaction products emit fluorescence. Therefore, by measuring the change in the fluorescence intensity derived from the MGO-protein reaction product due to the addition of the plant extract, the inhibition rate of the production of the MGO-protein reaction product can be calculated, and the glycation reaction inhibition rate derived from MGO Can be calculated. Plant extracts include those that emit fluorescence and those that absorb light and those that adsorb to proteins. Therefore, the influence of the component that does not adsorb to the MGO-protein reaction product is removed by precipitating the MGO-protein reaction product with trichloroacetic acid, and for the adsorbed component, the quenching rate derived from the plant extract is measured, and the apparent inhibition rate is determined. The true inhibition rate can be calculated by reducing the extinction rate.

[Saccharification reaction inhibition test]
(1) Apparent inhibition rate calculation Plant extract of water extraction or hot water extraction shown in Table 1 below is dissolved in 0.1M phosphate buffer (pH 7.4) so as to be 40 mg / mL. An extract solution was obtained. The plant extract of ethanol extraction was used as it was. 50 μL of 60 mM methylglyoxal (MGO), 50 μL of 20 mg / mL bovine serum albumin (BSA), a plant extract solution or a plant extract, and the plant extract having a final concentration (1.7 mg / mL, 3 mg 50 μL was added so as to be 3 mg / mL, 6.7 mg / mL, 13.3 mg / mL, or 3.3 v / v%), and the reaction was allowed to stand at a temperature of 37 ° C. for 1 week. After completion of the reaction, 17 μL of 100% (w / v) trichloroacetic acid solution was added and stirred, followed by centrifugation (rotation speed 1800 rpm, 10 minutes). Thereafter, the supernatant was removed by aspiration, washed with 0.1 M phosphate buffer (pH 7.4), and the phosphate buffer was removed by aspiration. Thereafter, 200 μL of alkaline phosphate buffer (pH 10.0) was added to the precipitate and dissolved, and this was used as a measurement sample.

  Moreover, what added 50 microliters of 0.1M phosphate buffer (pH7.4) instead of the sample plant extract solution or the plant extract was made into the control | contrast thing.

Using a fluorescence plate reader (device name: ARVO-MX, manufactured by PerkinElmer), the excitation filter is 355/40 nm, the fluorescence filter is 460/25 nm, the fluorescence intensity (Is) of each measurement sample, and the fluorescence intensity of the blank ( Ib), and the fluorescence intensity (Ic) of the control are measured and the following formula 1:
Apparent inhibition rate (%) = 100 − {(Is−Ib) / (Ic−Ib)} × 100
Based on this, the apparent inhibition rate was calculated.

(2) Extinction rate calculation 50 μL of 60 mM methylglyoxal (MGO) prepared with this buffer solution and 50 μL of 20 mg / mL bovine serum albumin (BSA) were mixed with 50 μL of 0.1 M phosphate buffer (pH 7.4). The reaction was allowed to stand at a temperature of 37 ° C. for 1 week. Thereafter, the plant extract solution or plant extract was subjected to the final concentration described in Table 1 (1.7 mg / mL, 3.3 mg / mL, 6.7 mg / mL, 13.3 mg / mL, or 3.3 v / v). %) And left standing for 1 day, 17 μL of a trichloroacetic acid solution was added and stirred, followed by centrifugation (rotation speed 1800 rpm, 10 minutes). Thereafter, the supernatant was removed by aspiration, washed with 0.1 M phosphate buffer (pH 7.4), and the phosphate buffer was removed by aspiration. Thereafter, 200 μL of alkaline phosphate buffer (pH 10.0) was added to the precipitate and dissolved, and this was used as a measurement sample.

  Moreover, what added 50 microliters of 0.1M phosphate buffer (pH7.4) instead of the sample plant extract solution or the plant extract was made into the control | contrast thing.

The fluorescence intensity (I ′s) of each measurement sample and the fluorescence intensity (I′c) of the control object are measured by the same method as in (1) above, and the following formula 2:
Extinction rate (%) = {(I'c-I's) / (I'c)} x 100
Thus, the extinction rate was calculated.

(3) Calculation of True Inhibition Rate For the true inhibition rate based on the fluorescence intensity of the MGO-BSA reaction product, the following equation that excludes the influence of fluorescence generation or fluorescence quenching derived from components other than the MGO-BSA reaction product: 3:
True inhibition rate (%) = Apparent inhibition rate (%)-Quenching rate (%)
Calculated by

[result]
The true inhibition rate of each plant extract measured / calculated above is shown in Table 1 below as the saccharification reaction inhibition rate.

  As shown in Table 1, each plant extract shown in Table 1 exhibited an MGO-BSA reaction product formation inhibitory action, that is, a saccharification reaction inhibitory action. In particular, Aojiso (leaf), rosemary (branch), savory (whole plant), Yerba mate (leaf), rose hip (whole plant), peanut (seed coat), grape (kyoho branch, fruit, pericarp, seed) , Grape (Delaware, branch), grape (Pione, branch), grape (Berry A, branch), grape (Campbell, seed), coffee tree (seed (roasted, raw)), apricot (seed), oyster tree (seed) , Olives (fruits, leaves), chanoki (leaves) are equivalent to or higher than the inhibition rate (about 30%) of 3 mM aminoguanidine, which is known to have inhibition of AGE generation and carbonyl compound scavenging. The saccharification reaction inhibitory action was excellent.

  In addition, the parts other than those shown in Table 1 for cypress, chestnut, peanut, oyster, rosehip, blueberry, pomegranate, rosemary, grape, apricot, perilla, sage, peppermint, savory, yerba mate, coffee, olives, tea Using the site, an experiment was performed to determine the saccharification reaction inhibition rate in the same manner as described above. In particular, the site shown in Table 1 showed an excellent saccharification reaction inhibitory action.

  The saccharification reaction inhibitor of this invention has the outstanding saccharification reaction inhibitory effect by containing a specific plant or its processed material. The saccharification reaction inhibitor of the present invention can be suitably used as it is or after adding various components and contained in a composition for oral administration or ingestion or a composition for external use on the skin.

Claims (3)

  One or more selected from the group consisting of Japanese cypress, chestnut, peanut, oyster, rosehip, blueberry, pomegranate, rosemary, grape, apricot, perilla, sage, peppermint, savory, yerba mate, coffee tree, olive, chanoki A saccharification reaction inhibitor containing a plant or a processed product thereof.   The saccharification reaction inhibitor according to claim 1, which inhibits a reaction between an intermediate in the saccharification reaction and an amino acid, peptide, protein or salt thereof.   The saccharification reaction inhibitor according to claim 1 or 2, wherein the intermediate is methylglyoxal.