TW202000220A - Glycation inhibitor - Google Patents

Abstract

To provide a novel glycation inhibitor. A glycation inhibitor which comprises as an active ingredient an extract of one or more plants selected from the plant group consisting of horseradish (Armoracia lapathifolia), Lonicera caerulea var. emphyllocalyx, Monarda fistulosa L., Ligularia japonica, Potentilla erecta, Penthorum chinense, Hibiscus syriacus and Bauhinia forficate L.

Description

Saccharification inhibitor

The invention relates to an inhibitor of saccharification reaction.

The saccharification reaction is a series of reactions in which the amine groups of proteins or amino acids and the carbonyl groups of reducing sugars such as glucose are non-enzymatically bonded as a starting point, including the pre-reaction of the formation of an amadori compound via a Schiff base; and After dehydration, oxidation, condensation, rearrangement and other reactions to produce saccharification end products (AGEs) later reaction. It is reported that the generated AGEs are one of the receptors of AGEs, and are expressed in the cell membranes of monocytes/macrophages, T cells, endothelial cells, fibroblasts, smooth muscle cells, nerve cells, glial cells, chondrocytes, etc. RAGE (Receptor for AGEs) binds to activate the RAGE message and activates NADPH (nicotinamide adenine dinucleotide phosphate, nicotinic adenine dinucleotide phosphate) oxidase, thereby generating reactive oxygen species , Causing an inflammatory response via the transcription factor NF-κB (for example, refer to Non-Patent Documents 1 and 2). In addition, it is believed that RAGE is localized in the vascular endothelium, so the activation of RAGE information is involved in the pathogenesis or development of diseases related to arteriosclerosis. In addition, it is also reported to participate in diabetic complications (such as diabetic nephropathy, diabetic retinopathy) , Diabetic neuropathy), hypertension, dementia, cancer, nonalcoholic steatohepatitis, infertility, obesity, male erectile dysfunction, periodontal disease, Alzheimer's disease, cataracts, osteoporosis, etc. Development (for example, refer to patent documents 1, 2 and non-patent documents and non-patent documents 3).

Generation of AGEs derived from saccharification intermediates such as glyoxal (GO), glycolaldehyde (GA), methylglyoxal (MGO), 3-deoxyglucosone, etc., or sugar metabolites such as glyceraldehyde (GLA) Also included in the saccharification reaction. It is known that these AGEs are produced much earlier than AGEs derived from glucose. Reports include GLAP (glyceraldehyde-derived pyridinium) derived from GLA, or MG-H1 (methylglyoxal-derived hydroimidazolone 1, derived from MGO), source derived from MGO. GA-pyridine from GA, GO, or CML (Nε-(carboxymethyl) lysine, Nε-(carboxymethyl) lysine) generated by oxidation pressure is combined with RAGE (for example, refer to non-patent literature 4～6). In addition, AGEs derived from GLA are called toxic AGEs (Toxic AGEs) (TAGE), suggesting that they are related to various diseases via RAGE (for example, refer to Non-Patent Document 7). The control of the glycation reaction in the living body can be a goal aimed at preventing, improving or treating the onset and development of lifestyle-related diseases or aging-related diseases. [Prior Technical Literature] [Patent Literature]

[Patent Document 1] Japanese Patent No. 3837494 [Patent Document 2] Japanese Patent No. 4143716 [Non-patent literature]

[Non-Patent Document 1] Folia Pharmacol. Jpn., 2014, Vol. 143, p. 10-13 [Non-Patent Document 2] Journal of Leukocyte Biology, 2013, Vol. 94, p. 55-68 [Non-Patent Document 3] MINERVA ENDOCRINOL, 2014, Vol. 39, p.167-74 [Non-Patent Document 4] Biosci Biotechnol Biochem., 2018, Vol. 82, p.312-319 [Non-Patent Literature 5] Biochemistry, 2014, Vol. 53, p. 3327-3335 [Non-Patent Document 6] J Biol Chem, 1999, Vol. 274, p. 31740-31749 [Non-Patent Document 7] Folia Pharmacol. Jpn, 2012, Vol. 139. p.193-197

[Problems to be solved by the invention]

The main purpose of the present invention is to provide a novel saccharification reaction inhibitor. [Technical means to solve the problem]

The inventors of the present invention have made intensive research in order to solve the above problems, and as a result, they have found that a specific plant extract has an excellent saccharification reaction inhibitory effect, thereby completing the present invention.

As the present invention, for example, the following can be mentioned. (1) An inhibitor of saccharification reaction (hereinafter, referred to as "inhibitor of the present invention"), which contains a member selected from the group consisting of Western wasabi, Lonicera japonicus, American mint, Ligularia grandis, Erectus paniculata, root vegetable, hibiscus and The extract of one or more plants in the flora composed of Bauhinia brasiliensis as the active ingredient. (2) A type of hypertension, dementia, cancer, nonalcoholic steatohepatitis, infertility, obesity, male erectile dysfunction, periodontal disease, Alzheimer's disease, cataract, osteoporosis prevention, improvement or The therapeutic composition contains the plant extract as described in (1) above. (3) A composition for the prevention, improvement or treatment of vascular diseases, which contains the plant extract as described in (1) above. (4) The composition for prevention, improvement or treatment of vascular diseases as described in (3) above, wherein the vascular diseases are inflammation of blood vessels, arteriosclerosis, diabetic vascular complications or vascular endothelial dysfunction. [Effect of invention]

The saccharification reaction inhibitor of the present invention has an excellent saccharification reaction inhibitory effect. In addition, the saccharification reaction inhibitor of the present invention contains a plant extract with edible experience and few side effects as an effective ingredient, and has high safety.

Plant extract The plant extract of the present invention is one species selected from the group consisting of Western wasabi, Lonicera japonicus, Lonicera sibiricum, Lithocarpus, Erectus paniculata, Rhizoma arvense, hibiscus, and Bauhinia Extracts of more than 2 types of plants. Western Horseradish is a plant of the horseradish genus Cruciferae, and its scientific name is Armoracia rusticana P. Gaertn., B. Mey. et Schreb. The alias is also known as horseradish, raifort, horseradish, horse wasabi, horse radish, or ezo wasabi. Lonicera caerulea L. is a plant of the Lonicera family of Loniceraceae. Its scientific name is Lonicera caerulea L. The alias is also called black blind fruit, blue fruit or blue indigo fruit. It is a plant of the genus American mint in the Labiatae family, and its scientific name is Monarda fistulosa L. The alias is also called horse mint.

Ligularia grandiflora is a plant belonging to the genus Ligularia in the family Asteraceae, and its scientific name is Ligularia japonica Less. The plant of the genus Potentilla erecta, the scientific name for Potentilla erecta (L.) Raeusch. The alias is also known as cephalopanthes or pheasant feast root. Root-root cuisine The root-root vegetable is a plant of the genus Rhizome, and its scientific name is Penthorum chinense Pursh. The alias is also called catch yellow grass. Hibiscus is a plant in the genus Hibiscus of the Malvaceae family, and its scientific name is Hibiscus syriacus L. Brazilian Bauhinia is a plant belonging to the genus Bauhinia in the legume family. Its scientific name is Bauhinia forficata Link. The alias is also known as pata de vaca.

The parts of the plant used in the production of the plant extract of the present invention are not particularly limited, and various parts of the above plants can be used. For example, flowers, spikes, peels, fruits, stems, leaves, branches, branches and leaves, stems, bark, rhizomes, root barks, roots, seeds, galls, heartwood, aerial parts, aerial parts, or whole grasses can be used. The production method of the plant extract of the present invention is not particularly limited, and can be produced by a commonly used method. For example, it can be manufactured by directly or cutting each part of the plant to an appropriate size for juice extraction or extraction with a solvent. As the extraction solvent, for example, water, various organic solvents, or a mixed solvent of these can be used. Examples of organic solvents used for extraction include lower alcohols (methanol, ethanol, etc.), chloroform, ethyl acetate, and n-hexane. Among the extraction solvents, water, methanol and ethanol are particularly preferred. In addition, these solvents may be used alone or in combination of two or more. The use amount of the extraction solvent varies according to the plant raw material or extraction solvent used, and is preferably within a range of 1:2 to 1:30 (plant raw material: extraction solvent) in terms of weight ratio, preferably 1:3 to Within the range of 1:20, it is more preferably within the range of 1:5 to 1:10. The extraction time is suitably in the range of 1 hour to 15 days. The extraction temperature is suitably in the range of 5 to 100°C. The extraction method is not particularly limited, and any method such as batch extraction and continuous extraction using a column can be applied.

The obtained plant extract can also be used in its original state, and can be further refined within a range that does not affect its activity if necessary. Such refining treatment may be carried out by a usual method, for example, it can be carried out by filtering plant extracts by a conventional method. Thereafter, the obtained filtrate can be concentrated under reduced pressure and freeze-dried to prepare the plant extract of the present invention. Composition of the invention

The glycation reaction inhibitor of the present invention, or hypertension, dementia, cancer, non-alcoholic steatohepatitis, infertility, obesity, male erectile dysfunction, periodontal disease, Alzheimer's disease, cataract, osteoporosis Or a composition for the prevention, improvement or treatment of vascular disease (hereinafter, collectively referred to as "the composition of the present invention") is one containing one or two or more plant extracts obtained as described above. The composition of the present invention can be produced, for example, by directly using the plant extract obtained as described above or mixing it with a material that can be used as a carrier, and then processing it into various forms such as powder, block, and liquid by conventional methods. . In the composition of the present invention, additives acceptable in medicine, cosmetics or food can be arbitrarily formulated. Examples of such additives include binders, disintegrants, lubricants, dispersants, suspending agents, emulsifiers, buffers, antioxidants, excipients, surfactants, ultraviolet inhibitors, and metal ion blockers. One, two or more of these may be used in appropriate amounts for agents, tackifiers, preservatives, antibacterial agents, moisturizers, and pigments.

The formulation amount of the plant extract in the composition of the present invention is not particularly limited, and it is preferably 0.0001% by weight (hereinafter, abbreviated as "%") in terms of solids in the total weight of the composition of the present invention, preferably It is 0.01% by weight or more, and more preferably 0.05 to 50%. The composition of the present invention can be suitably prepared into tablets, powders, granules, capsules, liquids, syrups, drops, conditioning liquids, lotions, ointments, creams and other dosage forms by conventional methods. The intake amount of the composition of the present invention varies according to the type, dosage form, administration method, age, body weight, symptoms, etc. of the plant extracts contained in it. Usually, adults take the weight of plant extracts per day, and it is suitable to set It is in the range of 0.1 to 100 g, preferably in the range of 1 to 60 g, but it is not necessarily limited to this range. This daily intake can be taken once, or it can be divided into multiple intakes. The use form of the composition of the present invention is not particularly limited, and as a pharmaceutical composition, food composition, cosmetic composition, etc., for example, it can be used in the form of oral preparations, food and drink, and external preparations.

In the case of using the composition of the present invention in the form of oral preparations, the plant extract of the present invention can be used directly or as required by appropriate preparation of the pharmaceutical or food additives used in the preparation of oral preparations and then used It is obtained by conventional formulation. Examples of such additives include excipients, fillers, extenders, binders, disintegrants, surfactants, seasonings, fragrances, and lubricants. In the preparation of oral preparations, other physiologically functional materials can also be formulated. The oral dosage form is not particularly limited, and examples include tablets, powders, granules, capsules, liquids, syrups, and drops. The content of the plant extract in the oral preparation may be appropriately prepared according to the type and dosage form of the plant extract, for example, as long as it is in the total amount of the preparation, calculated as solid content, it is preferably 0.5 to 60%. The plant extract can be formulated within a range of 3-50%, particularly preferably 5-10%.

When the composition of the present invention is used in the form of an external preparation, the plant extract of the present invention can be directly or as required to prepare appropriate additives that are pharmaceutically or cosmetically acceptable when used in the manufacture of external preparations. Obtained by the method of preparation. Examples of such additives include base materials for ointments, alcohols, polyols, water-soluble polymers, antioxidants, pH adjusters, ultraviolet inhibitors, metal ion blocking agents, tackifiers, surfactants, and purifications. Water, preservatives, antibacterial agents, oils, higher fatty acids, fatty acid esters, humectants, pigments, vitamins, amino acids, etc. Furthermore, the composition of the present invention can be used in the form of external preparations in known quasi-drugs and cosmetics. In the external preparation, in addition to the composition of the present invention, components for enhancing the inhibitory effect of the saccharification reaction may be further prepared, or other physiologically functional materials may be prepared. Examples of such components include ultraviolet inhibitors, antioxidants, humectants, cell activators, and blood flow promoters. One or more of these may be used in appropriate amounts. The specific use of the external preparation is not particularly limited, and examples thereof include lotions, emulsions, creams, ointments, lotions, oils, and compresses. The content of the plant extract in the external preparation may be appropriately prepared according to the type and dosage form of the plant extract of the present invention contained, for example, as long as it is in the total amount of the preparation, calculated in terms of solid content, from 0.0001 to 30 %, preferably 0.001 to 25%, and particularly preferably 0.5 to 20%.

When the composition of the present invention is used in the form of food and drink, it can be obtained by mixing the plant extract of the present invention with a well-known food material or food and drink and processing it into food by a conventional method. The form of such food and drink is not particularly limited, and examples thereof include powder, block, liquid, syrup, and jelly. In food and beverage, other ingredients allowed in food can be blended. Examples of such components include nutrients, excipients, bulking agents, sweeteners, flavors, colorants, preservatives, emulsifiers, cosolvents, polyols, organic acids, inorganic acids, and high water solubility. molecular. One or more of these may be used in appropriate amounts. Examples of the types of food and drink include beverages, powdered beverages, home-cooked dishes, noodles, toasted bread, bread, snacks, and seasonings. The amount of the plant extract added to the food and drink varies according to the presence or absence of additives or the type of food. Based on the conversion of the solid content of the plant extract contained, it is preferably set within the range of 0.01 to 50%, preferably It is set within the range of 0.1 to 30%, but it is not necessarily limited to this range.

The "glycation reaction" in this specification is a series of reactions in which amine groups of proteins or amino acids react with carbonyl groups of reducing sugars to form AGEs, and is also known as Mena reaction. AGEs are produced not only from glucose or fructose, but also from saccharification intermediates or sugar metabolites.

The "saccharification reaction inhibitor" of the present invention is a composition having an effect of inhibiting the saccharification reaction and inhibiting the generation of AGEs. The saccharification reaction inhibitor of the present invention can suppress the generation of AGEs generated by the saccharification reaction of saccharification reaction intermediates or sugar metabolites and proteins, and prevent, improve or treat various phenomena generated by the saccharification reaction, so it can be used for example Knowing that the elasticity of cartilage involved in the glycation reaction is reduced, the aging of the skin (the formation of cross-linking of collagen that causes the reduction of the elasticity of the skin, wrinkles or sagging, the pigmentation that causes the dullness of the skin, etc.), arteriosclerosis, Diabetic complications (e.g. myocardial infarction, diabetic nephropathy, diabetic retinopathy, diabetic neuropathy), inflammation, cancer metastasis, hypertension, dementia, cancer, nonalcoholic steatohepatitis, infertility, obesity, male erection Prevention, improvement or treatment of dysfunction, periodontal disease, Alzheimer's disease, cataract, osteoporosis or vascular disease (eg inflammation of blood vessels, arteriosclerosis, diabetic vascular complications, vascular endothelial dysfunction). [Example]

Hereinafter, Examples, Test Examples, and Production Examples will be disclosed to further detail the present invention. However, the present invention is not limited to the scope shown in the following examples.

Example 1 Manufacture of plant extracts The plants Nos. 1 and 3 to 8 described in Table 1 were air-dried and then dried at 60°C overnight. After pulverized by a blender, 100 g of the obtained pulverized product was immersed in 500 mL of methanol, and allowed to stand at room temperature for one week. The filtrate obtained by filtering the obtained extract was distilled to remove methanol by a rotary evaporator to obtain extracts of plants of Nos. 1 and 3 to 8. Each of the obtained plant extracts (numbers 1 and 3 to 8 in Table 1) was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 50 mg/mL to prepare a specimen. The plant of No. 2 described in Table 1 thawed the frozen fruit and separated it into a solid part and a liquid part. 100 g of the solid part was immersed in 1 L of 70% aqueous ethanol and allowed to stand at room temperature for 1 hour. In the filtrate and the residue obtained by filtering the obtained extract, the residue was further immersed in a 10-fold amount of 70% aqueous ethanol and allowed to stand at 85°C for 1 hour. The filtrate obtained by filtering the obtained extract, the liquid portion after thawing, and the filtrate obtained by initial filtration are purified by a styrene-divinylbenzene series column, and the obtained is concentrated 1. Spray drying to obtain extract. The obtained extract (No. 2 in Table 1) was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 10 mg/mL to prepare a specimen. [Table 1]

Test Example 1 Measurement of inhibitory activity of AGEs derived from glyceraldehyde (GLA) (1) Test method After diluting each sample obtained in Example 1 with DMSO at a final concentration of 10, 100, and 500 μg/mL, each sample was 25 μL and 50 mg/mL bovine serum albumin (Sigma) Manufacturing) 50 μL, 100 mM DL-glyceraldehyde (manufactured by Wako Pure Chemical Industries, Ltd.) (hereinafter, referred to as "GLA solution") 25 μL, 1 M phosphate buffer (pH 7.4) 50 μL, ultra pure water 100 μL The mixture was mixed and reacted at 96°C for 24 hours at 96°C (manufactured by IWAKI) to obtain a reaction solution. As a positive control, aminoguanidine was used instead of the specimen. The obtained reaction liquid was measured using a microdisk reader (manufactured by Corona Electric Co., SH-9000), and the fluorescence intensity at an excitation wavelength of 370 nm and a fluorescence wavelength of 440 nm was measured. In addition, in the composition of the above reaction solution, DMSO was added instead of the sample, and ultrapure water was added instead of the GLA solution to prepare a reaction solution without adding the GLA solution (blank sample 1), and a reaction solution without only adding the sample (sample- ). The reaction solution (blank sample 2) with neither sample nor GLA solution added, the fluorescence intensity is measured in the same manner as above. Using the obtained fluorescence intensity value, the inhibitory activity rate (%) of the saccharification reaction was calculated by the following calculation formula.

[Number 1] The inhibitory activity rate of saccharification reaction (%) = [1－{(sample (+)－blank sample 1)/(sample (-)－blank sample 2)}]×100 The inhibitory activity rate was set as the vertical axis, and the sample concentration (μg/mL) was set as the formula of the logarithmic approximate curve plotted on the horizontal axis, and the 50% inhibitory concentration (IC ₅₀ ) (ppm) was calculated. The results are shown in Table 2.

(2) Results According to Table 2, it is shown that the plant extract of the present invention exhibits inhibition of AGEs derived from GLA (hereinafter, also referred to as “GLA- The activity of AGEs") has an excellent saccharification inhibition effect.

Test Example 2. Inhibition measurement of generation of AGEs derived from methylglyoxal (MGO) (1) Test method Each sample obtained in Example 1 was diluted with DMSO so that the final concentration became 10, 100, and 500 μg/mL. Each sample 25 μL and 50 mg/mL bovine serum albumin (manufactured by Sigma) 50 μL, 100 mM methylglyoxal solution (manufactured by Sigma) (hereinafter referred to as “MGO solution”) 25 μL, 1 50 μL of M phosphate buffer solution (pH 7.4) and 100 μL of ultrapure water were mixed, and reacted at 37° C. for 72 hours in a 96-well plate (manufactured by IWAKI) to obtain a reaction solution. As a positive control, aminoguanidine was used instead of the specimen. The obtained reaction liquid was measured using a microdisk reader (manufactured by Corona Electric Co., SH-9000), and the fluorescence intensity at an excitation wavelength of 370 nm and a fluorescence wavelength of 440 nm was measured. In addition, to the reaction solution of the above composition, add DMSO instead of the sample, add ultrapure water instead of the MGO solution to prepare a reaction solution without adding the MGO solution (blank sample 1), and a reaction solution without only adding the sample (sample- ). The reaction solution (blank sample 2) with neither specimen nor MGO solution is added, and the fluorescence intensity is measured in the same manner as above. Using the obtained fluorescence intensity value, the inhibitory activity rate (%) of the saccharification reaction was calculated by the following calculation formula.

[Number 2] Saccharification reaction inhibitory activity rate (%) = [1-{(sample (+)-blank sample 1)/(sample (-)-blank sample 2)}] × 100 According to the inhibition to be obtained The activity rate was set as the vertical axis, and the sample concentration (μg/mL) was set as the formula of the logarithmic approximate curve plotted on the horizontal axis, and the 50% inhibitory concentration (IC ₅₀ ) (ppm) was calculated. The results are shown in Table 2.

(2) Results It can be seen from Table 2 that the plant extract of the present invention exhibits an activity of inhibiting the generation of MGO-derived AGEs (hereinafter, also referred to as "MGO-AGEs"), and has an excellent saccharification reaction inhibitory effect.

實施例2 植物萃取物之製造 將表1中所記載之編號5及6之各植物進行風乾後，於60℃下乾燥一晚。藉由摻合機進行粉碎後，將所獲得之粉碎物100 g浸漬於水或乙醇500 mL中，於80℃下靜置1小時。對所獲得之萃取液進行過濾所獲得之濾液係藉由旋轉蒸發器蒸餾去除溶劑，獲得編號5及6之各植物之萃取物。 所獲得之各植物萃取物(表1之編號5及6)分別以50 mg/mL之濃度溶解於二甲基亞碸(DMSO)中，製成檢體。 試驗例3 源自GLA、MGO及乙醇醛(GA)之AGEs生成抑制活性測定 (1)試驗方法 將實施例2中所獲得之各檢體以最終濃度成為500 μg/mL之方式藉由DMSO進行稀釋。GLA-AGEs、MGO-AGEs之生成抑制活性係藉由與試驗例1及2之記載方法相同之方法進行測定。 源自GA之AGEs(以下，亦稱為「GA-AGEs」)之生成抑制活性係藉由以下之方法進行測定。將各檢體25 μL與100 mg/mL牛血清白蛋白(Sigma公司製造)50 μL、30 mM乙醇醛二聚物(Sigma公司製造)(以下，稱為「GA溶液」)25 μL、1 M磷酸緩衝液(pH值7.4)25 μL、超純水125 μL進行混合，藉由96孔板(IWAKI公司製造)於37℃下反應24小時，獲得反應液。作為陽性對照，使用胺基胍代替檢體。 所獲得之反應液係使用微盤讀取器(Corona Electric公司製造，SH-9000)，測定激發波長370 nm、螢光波長440 nm之螢光強度。又，於上述反應液之組成中，分別添加DMSO代替檢體、添加超純水代替GA溶液製備不添加GA溶液之反應液(空白樣品1)、僅不添加檢體之反應液(檢體-)、既不添加檢體亦不添加GA溶液之反應液(空白樣品2)，以與上述相同之方式測定螢光強度。 使用所獲得之螢光強度之值，藉由以下之計算式算出糖化反應之抑制活性率(%)。[Table 2]

Example 2 Manufacture of plant extracts Each plant No. 5 and No. 6 described in Table 1 was air-dried, and dried overnight at 60°C. After pulverization by a blender, 100 g of the obtained pulverized product was immersed in 500 mL of water or ethanol, and allowed to stand at 80°C for 1 hour. The filtrate obtained by filtering the obtained extract was distilled off the solvent by a rotary evaporator to obtain extracts of each plant No. 5 and 6. Each of the obtained plant extracts (numbers 5 and 6 in Table 1) was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 50 mg/mL to prepare a specimen. Test Example 3 Measurement of AGEs production inhibitory activity derived from GLA, MGO and glycolaldehyde (GA) (1) Test method Each sample obtained in Example 2 was carried out by DMSO in such a way that the final concentration became 500 μg/mL dilution. The production inhibitory activity of GLA-AGEs and MGO-AGEs was measured by the same method as described in Test Examples 1 and 2. The generation inhibitory activity of GA-derived AGEs (hereinafter, also referred to as "GA-AGEs") was measured by the following method. Each sample 25 μL and 100 mg/mL bovine serum albumin (manufactured by Sigma) 50 μL, 30 mM glycolaldehyde dimer (manufactured by Sigma) (hereinafter, referred to as “GA solution”) 25 μL, 1 M 25 μL of phosphate buffer (pH 7.4) and 125 μL of ultrapure water were mixed and reacted at 37° C. for 24 hours in a 96-well plate (manufactured by IWAKI) to obtain a reaction solution. As a positive control, aminoguanidine was used instead of the specimen. The obtained reaction solution was measured using a microdisk reader (manufactured by Corona Electric, SH-9000), and the fluorescence intensity at an excitation wavelength of 370 nm and a fluorescence wavelength of 440 nm was measured. In addition, in the composition of the above reaction solution, DMSO was added instead of the sample, and ultrapure water was added instead of the GA solution to prepare a reaction solution without adding the GA solution (blank sample 1), and a reaction solution without only adding the sample (sample- ). The reaction solution (blank sample 2) with neither sample nor GA solution added, the fluorescence intensity was measured in the same manner as above. Using the obtained fluorescence intensity value, the inhibitory activity rate (%) of the saccharification reaction was calculated by the following calculation formula.

[Number 3] Inhibition activity rate of saccharification reaction (%) = [1-{(sample (+)-blank sample 1) / (sample (-)-blank sample 2)}] × 100 The results are shown in the table 3. (2) The results can be seen from Table 3, showing that the water and ethanol extracts of Eupatorium arvense, Root Vegetables show the activity of inhibiting the formation of GLA-AGEs, MGO-AGEs and GA-AGEs, and have excellent saccharification reaction inhibitory effect . Test Example 4 Measurement of carboxymethyl lysine acid (CML) production inhibitory activity (1) Test method Each sample obtained in Example 2 was diluted with DMSO so that the final concentration became 1 μg/mL, and then Each sample 5 μL and 4 mg/mL bovine serum albumin (manufactured by Sigma) 50 μL, 2 M glucose (manufactured by Nacalai Tesque) 25 μL, 100 mM phosphate buffer (pH 7.4) 125 μL, ultrapure water 45 μL was mixed, and a 96-well plate (manufactured by IWAKI) was reacted at 60° C. for 24 hours to obtain a reaction solution. As a positive control, aminoguanidine was used instead of the specimen. Next, add 0.4 μg/mL of CML-BSA/Nε-(carboxymethyl) lysine-BSA (manufactured by CycLex) to 100 μL/well in an ELISA (enzyme linked immunosorbent assay). After incubating at 37°C for 1 hour for solid phase, wash with 0.05% Tween-20 phosphate buffer. Thereafter, 1% Block Ace (manufactured by Megmilk Snow Brand) was added at 200 μL/well, blocked at 37° C. for 1 hour, and washed with 0.05% Tween-20-containing phosphate buffer. Furthermore, 0.05 μg/mL anti-Nε-(carboxymethyl)lysine (Anti Nε-(carboxymethyl)lysine) (manufactured by Cosmo Bio) 50 μL/well and CML-BSA (for the calibration curve) or the above reaction solution are added The 50 μL/well mixed solution was allowed to stand at room temperature for 1 hour. After washing with 0.05% phosphate buffer containing Tween-20, add 5000-fold diluted anti-mouse IgG (γ-chain specific)-goat peroxidase antibody (Anti-Mouse IgG( γ-chain specific)-peroxidase antibody produced in goat) (manufactured by Sigma), and allowed to stand at room temperature for 1 hour. After washing with 0.05% Tween-20 phosphate buffer, add TMB solution (manufactured by Nacalai Tesque) at 100 μL/well, add 1 N sulfuric acid at 100 μL/well 10 minutes later, and use a microdisk reader ( Corona Electric Corporation, SH-9000) measured the absorbance at 450 nm. The amount of CML in each reaction solution was calculated from the calibration curve, and the amount of CML in the reaction solution to which no sample was added was set to 100%, and the CML production inhibition rate (%) caused by the added sample was calculated. (2) The results can be seen from Table 3, showing that the water and ethanol extracts of Rhizopus chinensis showed a higher degree of activity in inhibiting the formation of CML than the aminoguanidine known as a powerful saccharification inhibitor. Excellent inhibition of saccharification reaction. [table 3]

Next, examples of production of oral preparations, external preparations, foods and beverages containing the inhibitor of the present invention are shown. Production Example 1 Preparation of oral preparations (lozenges) The ingredients are mixed according to the following mixing ratio, and tablets are manufactured according to a conventional method.

製造例2 經口劑(顆粒劑)之製備 按照以下之調配比率混合各成分，按照常規方法製造顆粒劑。[table 3]

Production Example 2 Preparation of oral preparations (granules) The ingredients were mixed according to the following formulation ratio, and granules were produced according to a conventional method.

製造例3 經口劑(軟膠囊劑)之製備 按照以下之調配比率混合各成分，按照常規方法製造軟膠囊劑。[Table 4]

Production Example 3 Preparation of oral preparations (soft capsules) The ingredients were mixed according to the following formulation ratio, and soft capsules were manufactured according to a conventional method.

製造例4 外用劑(乳霜)之製備 按照以下之調配比率混合各成分，按照常規方法製造乳霜。[table 5]

Production Example 4 Preparation of external preparation (cream) The ingredients were mixed according to the following formulation ratio, and a cream was produced according to a conventional method.

製造例5 外用劑(化妝水)之製備 按照以下之調配比率混合各成分，按照常規方法製造化妝水。[Table 6]

Production Example 5 Preparation of external preparation (lotion) The ingredients were mixed according to the following formulation ratio, and a lotion was manufactured according to a conventional method.

製造例6 飲食品(飲料)之製備 按照以下之調配比率混合各成分，按照常規方法製造飲料。[Table 7]

Production Example 6 Preparation of Food and Beverage (Beverage) The ingredients were mixed according to the following mixing ratio, and the beverage was produced according to a conventional method.

製造例7 飲食品(糖果)之製備 按照以下之調配比率混合各成分，按照常規方法製造糖果。[Table 8]

Production Example 7 Preparation of Food and Beverage (Candy) The ingredients were mixed according to the following mixing ratio, and the candy was manufactured according to a conventional method.

[Table 9]

Claims (4)

An inhibitor of saccharification reaction, which contains one or two species selected from the group consisting of Western wasabi, Lonicera japonicus, Lonicera sibiricum, Ligularia grandiflora, Erectus paniculata, Rhizoma chinensis, Hibiscus, and Bauhinia The above plant extracts are used as active ingredients. A composition for the prevention, improvement or treatment of dementia, cancer, non-alcoholic steatohepatitis, infertility, obesity, male erectile dysfunction, periodontal disease, Alzheimer's disease, cataract or osteoporosis, which contains Plant extract as in claim 1. A composition for the prevention, improvement or treatment of vascular diseases, which contains the plant extract according to claim 1. The composition for prevention, improvement or treatment of vascular disease according to claim 3, wherein the vascular disease is inflammation of blood vessels, arteriosclerosis, diabetic vascular complications or vascular endothelial dysfunction.