JP7007813B2 - Oxidizing proteolytic enzyme activity enhancer and glycation stress inhibitor - Google Patents

Description

The present disclosure relates to an oxidative proteolytic enzyme activity enhancer, a saccharification stress inhibitor, and a beverage composition, a food composition, a functional food, a cosmetic, a supplement, a quasi-drug or a pharmaceutical product containing them.

Oxidation and glycation reactions in vivo are known to have adverse effects on cells and tissues. The production and accumulation of oxidized and glycated proteins cause diabetic complications, Alzheimer's disease, cataracts, arteriosclerosis and other diseases, and aging and functional deterioration of tissues such as skin. Normally, these proteins are degraded and removed by proteolytic enzymes such as Oxidized Protein hydride (OPH) and proteasomes, but it is known that the activity of these enzymes decreases with aging.
Extracts of plants such as Roman chamomile, Houttuynia cordata, hawthorn, grape and Hishi have been proposed as substances for enhancing OPH activity (Patent Documents 1 and 2).

"Glycation stress" is a concept that comprehensively captures the stress on the living body due to the loading of reducing sugars and aldehydes, and the subsequent reaction. Abnormal proteins produced by glycation stress are attracting attention as being associated with diseases such as lifestyle-related diseases and aging.
The abnormal protein is generally a protein that has undergone oxidation, saccharification, aldehyde modification, or the like due to aging or the like. Accumulation of these in the body causes functional deterioration and inflammation of functional proteins, resulting in diabetic complications, Alzheimer's disease, cataracts, arteriosclerosis and other diseases, as well as aging and function of tissues such as skin. It is believed to cause a decline.
The Maillard reaction in the living body is considered to be one of the causes of the production of abnormal proteins. For example, by a glycation reaction (Maillard reaction) in the living body, proteins such as collagen are saccharified and amino acids in the proteins become AGEs (advanced glycation end products), or a crosslinked structure is formed between proteins by AGEs. , Becomes an abnormal protein.

N-phenacylthiazolium bromide (PTB) and N-phenacyl-4,5-dimethylthiazolium bromide (ALT-711) are known as agents for degrading the cross-linked structure between proteins by AGEs (Non-Patent Documents 1 and 2). However, they have not been utilized in terms of stability and side effects. In addition, lotus leaf extract, red pepper extract and the like have been studied (Patent Document 3).

WO2011 / 004733 WO2014 / 126199 JP-A-2007-119373

Sara Vasan et al. Nature, 382, p275-278 (1996) Sara Vasan et al. Archives of Biochemistry and Biophysics, 419, p89-96 (2003)

In AGEs and ALEs (advanced glycation end products of lipids), reactive groups such as reducing sugars such as glucose and aldehyde groups and ketone groups due to β-oxidation and peroxidation of lipids react with proteins or lipids in the living body (Maillard). It is produced by reacting). It has become clear that the dysfunction of functional proteins and the activation of AGEs receptors occur when these are produced and accumulated in vivo. These are considered as one of the factors that cause functional deterioration, inflammation, lifestyle-related diseases, aging, etc., and the concept that comprehensively captures these series of phenomena and symptoms is called saccharification stress or carbonyl stress. There is.

In addition, when the functional protein is saccharified by the Maillard reaction and causes dysfunction, it is thought that active oxygen is excessively generated and the scavenging action of active oxygen in the living body does not function sufficiently, resulting in oxidative stress. Has been done. Like glycation stress, oxidative stress is considered to be one of the causes of various diseases including lifestyle-related diseases and aging.

From the viewpoint of anti-aging and prevention of lifestyle-related diseases, there is a demand for new substances and methods capable of suppressing, decomposing or removing the above-mentioned abnormal proteins and suppressing or reducing glycation stress and oxidative stress.

In addition, there are high expectations for the possibility of treating and preventing the disease, aging of the tissue, and functional deterioration of the tissue by enhancing the activity of enzymes such as OPH.

Therefore, the present disclosure provides, in one embodiment, a new substance capable of enhancing OPH activity and a new substance capable of suppressing or reducing glycation stress.

The present disclosure relates to, in one embodiment, an oxidative proteolytic enzyme activity enhancer comprising an extract of fenugreek as an active ingredient.

The present disclosure relates to, in other embodiments, an oxidative proteolytic enzyme activity enhancer comprising an extract of one or more plants selected from the group consisting of fenugreek, fennel and hibiscus as an active ingredient.

The present disclosure relates to, as another aspect, a glycation stress inhibitor containing fenugreek and fenugreek extracts as active ingredients.

The present disclosure, in other aspects, is a beverage composition, food composition, functional food, cosmetics, supplement, quasi-drug or pharmaceutical product containing fenugreek and fenugreek extract as an active ingredient for suppressing saccharification stress. Regarding.

In other embodiments, the present disclosure comprises a beverage composition, a food composition, a functional food, a cosmetic product, which comprises at least one of the oxidative proteolytic enzyme activity enhancer of the present disclosure and the saccharification stress inhibitor of the present disclosure as an active ingredient. Regarding supplements, quasi-drugs or pharmaceutical products.

According to the present disclosure, in one embodiment, it is possible to provide a new OPH activity enhancer and a beverage composition, a food composition, a functional food, a cosmetic, a supplement, a quasi drug or a pharmaceutical product capable of enhancing the OPH activity.

According to the present disclosure, in one embodiment, it is possible to provide a new saccharification stress inhibitor and a beverage composition, a food composition, a functional food, a cosmetic, a supplement, a quasi-drug or a pharmaceutical product capable of suppressing the saccharification stress.

[OPH activity enhancer]
In the present disclosure, water extracts of fenugreek (scientific name: Trigonella foenum-glaecum), fenugreek (scientific name: Foenicularum vulgare) and hibiscus (scientific name: Hibiscus sabdarifa), particularly hot water extracts, are known as OPH. Based on the new finding that the enzyme activity of (AARE) can be enhanced.

It is known that OPH is widely distributed in the living body and preferentially decomposes substances caused by glycation stress and oxidative stress in the living body such as oxidative protein and glycated protein. Therefore, by enhancing the enzymatic activity of OPH, it is expected that the decomposition efficiency of the oxidized protein and the glycated protein can be improved, thereby suppressing or reducing the glycated stress and the oxidative stress. According to the OPH activity enhancer of the present disclosure, since the enzyme activity of OPH can be enhanced, the oxidized protein can be decomposed and removed or the accumulation of the oxidized protein can be suppressed in one or more embodiments. According to the OPH activity enhancer of the present disclosure, glycation stress and / or oxidative stress can be suppressed or reduced in one or more embodiments. According to the OPH activity enhancer of the present disclosure, in one or more embodiments, it is possible to prevent, improve, or treat diseases, tissue aging, and tissue dysfunction caused by glycation stress or oxidative stress. .. According to the OPH activity enhancer of the present disclosure, metabolism can be improved in one or more embodiments.

The present disclosure relates to, in one aspect, an OPH activity enhancer comprising an extract of fenugreek as an active ingredient. The OPH activity enhancer of the present disclosure may further comprise an extract of Fennell in one or more embodiments. The OPH activity enhancer of the present disclosure may further comprise an extract of Shoeblackplant in one or more embodiments. The OPH activity enhancer of the present disclosure comprises, in one or more embodiments, an extract of fenugreek and at least one extract of fenugreek and hibiscus as an active ingredient. The present disclosure also relates to OPH activity enhancers, in other embodiments, comprising an extract of one or more plants selected from the group consisting of fenugreek, fenugreek and hibiscus as an active ingredient.

The OPH activity enhancer of the present disclosure can be used to suppress glycation stress in one or more embodiments. The OPH activity enhancer of the present disclosure can be used in one or more embodiments for the prevention, amelioration, or treatment of diseases, tissue aging, and tissue dysfunction caused by glycation stress or oxidative stress. The OPH activity enhancers of the present disclosure can be used in one or more embodiments for anti-aging or improving metabolism.

When the OPH activity enhancers of the present disclosure contain extracts of plants other than fenugreek, they may be blended in the same ratio (1 part by weight to 1 part by weight of the extract of fenugreek) or in different ratios. It may be blended with.

The OPH activity enhancers of the present disclosure include, in one or more embodiments, an extract of fenugreek and an extract of fenugreek. In this embodiment, the blending ratio (dry weight ratio) of the fenugreek extract and the fenugreek extract is 0.1 to 10 parts by weight of the fenugreek extract per 1 part by weight of the fenugreek extract, and the OPH activity. From the point of view of enhancement, the extract of fenugreek is 0.2-5 parts by weight.

The OPH activity enhancer of the present disclosure comprises, in one or more embodiments, an extract of fenugreek and an extract of hibiscus. In this embodiment, the blending ratio (dry weight ratio) of the fenugreek extract and the hibiscus extract is 0.1 to 10 parts by weight of the hibiscus extract per 1 part by weight of the fenugreek extract, and the OPH activity. From the point of view of enhancement, the hibiscus extract is 0.2-5 parts by weight.

The OPH activity enhancers of the present disclosure include, in one or more embodiments, an extract of fenugreek, an extract of fenugreek and an extract of hibiscus. In this embodiment, the blending ratio (dry weight ratio) of the phenuglique extract, the phenell extract and the hibiscus extract is 0.05 to 20 parts by weight per part by weight of the phenuglique extract. , The hibiscus extract is 0.05 to 20 parts by weight, and from the viewpoint of enhancing OPH activity, the phenel extract is 0.1 to 10 parts by weight, and the hibiscus extract is 0.1 to 10 parts by weight. be.

The extract may be extracted from any part. Examples of the site of use include whole plants, flowers, leaves, seeds, fruits, branches, stems, roots, calyxes, bracts and the like in one or more embodiments. The site to be used may be one type, or two or more types of sites may be used in combination. Fenugreek is preferably seeds in one or more embodiments. Phenels are preferably seeds in one or more embodiments. The hibiscus is preferably a shoeblackplant and a bract in one or more embodiments.

The extraction method is not particularly limited, and examples thereof include solvent extraction and the like in one or more embodiments. Examples of the extraction solvent include water, alcohols such as methanol and ethanol, ether and the like in one or more embodiments. Examples of the extract include a water extract, an organic solvent extract, and the like in one or more embodiments. Examples of the water extract include hot water extracts and the like in one or more embodiments. The temperature of the solvent extraction may be 60 to 100 ° C., 70 to 90 ° C., 80 ° C. or the like in one or more embodiments. In one or more embodiments, the solvent extraction time may be 30 minutes to 5 hours, 1 to 5 hours, 2 to 5 hours, 3 to 5 hours, 1 hour, or the like. The temperature of hot water extraction may be 60 to 100 ° C., 70 to 90 ° C., 80 ° C. or the like in one or more embodiments. In one or more embodiments, the hot water extraction time may be 30 minutes to 5 hours, 1 to 5 hours, 2 to 5 hours, 3 to 5 hours, 1 hour, or the like. The form of the extract may be an aqueous solution, a concentrated solution, a dried product or the like in one or more embodiments. Drying can be performed in one or more embodiments by heat-treating the extract obtained by solvent extraction. The heating temperature is 70-150 ° C or 110 ° C in one or more embodiments. The heating time is 1-10 hours or 4 hours in one or more embodiments.
Suitable methods for the production of the Maillard reaction product degrading agent of the present disclosure are as follows in one or more embodiments: An appropriate amount of phenuglique powder (eg, 2 g) and fennel powder (eg, 2 g) are extracted with an appropriate amount of distilled water (eg, 40 mL) at 80 ° C. for 1 hour, then cooled to room temperature, and then the obtained slurry is obtained. After filtration, the resulting filtrate is used as a plant extract. When using dried plant extracts, the resulting plant extracts are placed on aluminum trays and dried, for example, in an incubator set at 110 ° C. for 4 hours.

The form of the OPH activity enhancer of the present disclosure is not particularly limited, and examples thereof include solids, granules, powders, pastes, and liquids in one or more embodiments.

In one or more embodiments, the OPH activity enhancer of the present disclosure may contain extracts of other foods and other ingredients as long as the active ingredient does not interfere with the OPH activity enhancer function.

The OPH activity enhancer of the present disclosure can be used as a raw material for beverage compositions, food compositions, functional foods, cosmetics, supplements, feeds, quasi-drugs, pharmaceuticals and the like in one or more embodiments. The form as a raw material is not particularly limited, and examples thereof include solids, granules, powders, pastes, and liquids in one or more embodiments.
The present disclosure relates, as one or more embodiments, to beverage compositions, food compositions, functional foods, cosmetics, supplements, feeds, quasi-drugs or pharmaceuticals comprising the OPH activity enhancers of the present disclosure.

The daily intake of the OPH activity enhancer of the present disclosure is not particularly limited, and in one or more embodiments, it is 5 mg or more, or 2000 mg or less.

[Manufacturing method of OPH activity enhancer]
The present disclosure relates to, in other embodiments, a method for producing an OPH activity enhancer for enhancing OPH activity (method for producing an OPH activity enhancer according to the present disclosure). The method for producing an OPH activity enhancer of the present disclosure comprises, in one or more embodiments, solvent extraction of fenugreek. The method for producing an OPH activity enhancer of the present disclosure may further comprise, in one or more embodiments, solvent extraction of one or more plants selected from the group consisting of phenel and hibiscus. ..
In addition, the method for producing an OPH activity enhancer according to the present disclosure comprises, in one or more embodiments, solvent extraction of one or more plants selected from the group consisting of fenugreek, phenel and hibiscus. The present invention relates to a method for producing an OPH activity enhancer for enhancing OPH activity. According to the method for producing an OPH activity enhancer of the present disclosure, the OPH activity enhancer of the present disclosure can be produced.

Solvent extraction may include, in one or more embodiments, solvent extraction of a dry plant.

The method for producing an OPH activity enhancer of the present disclosure may include, in one or more embodiments, filtering or centrifuging the extract obtained by solvent extraction.

The method for producing an OPH activity enhancer of the present disclosure may include drying the obtained extract in one or more embodiments.

The production method of the present disclosure may include, in one or more embodiments, mixing an extract of fenugreek with an extract of a plant other than fenugreek to obtain a mixture of the extracts. The mixing ratio may be the same ratio (1 part by weight with respect to 1 part by weight of the fenugreek extract) or different ratios in one or more embodiments.

The production method of the present disclosure comprises mixing these extracts in one or more embodiments so that the extract of fenugreek is 0.1-10 parts by weight per part by weight of the extract of fenugreek. However, from the viewpoint of producing an OPH activity enhancer capable of further enhancing OPH activity, it may be included to mix these extracts so that the extract of fenugreek is 0.2 to 5 parts by weight. good.

The production method of the present disclosure comprises mixing these extracts in one or more embodiments so that the hibiscus extract is 0.1-10 parts by weight per 1 part by weight of the fenugreek extract. However, from the viewpoint of producing an OPH activity enhancer capable of further enhancing OPH activity, it may be included to mix these extracts so that the hibiscus extract is 0.2 to 5 parts by weight. good.

In one or more embodiments, the production method of the present disclosure comprises 0.05 to 20 parts by weight of the phenus extract and 0.05 to 20 parts by weight of the hibiscus extract per 1 part by weight of the phenuglique extract. It may be included to mix these extracts so as to be, and from the viewpoint of producing an OPH activity enhancer capable of further enhancing OPH activity, the phenel extract is 0.1 to 10 parts by weight, and the hibiscus is extracted. It may include mixing these extracts so that the product is 0.1 to 10 parts by weight.

The production method of the present disclosure may include, in one or more embodiments, mixing fenugreek with a plant other than fenugreek and solvent-extracting the resulting mixture to obtain a mixed extract. The mixing ratio may be the same ratio (1 part by weight with respect to 1 part by weight of fenugreek) or different ratios in one or more embodiments. Further, the mixture may be mixed so that the ratio of the extract of each plant in the obtained mixed extract is the same ratio (1 part by weight with respect to 1 part by weight of the extract of fenugreek), or the ratio is different. May be mixed with.

[Method for enhancing OPH activity]
The present disclosure relates to a method for enhancing OPH activity, which comprises administering an extract of fenugreek to a living body in one embodiment. The present disclosure relates to a method of enhancing OPH activity comprising administering to a living body an extract of one or more plants selected from the group consisting of fenugreek, fenugreek and hibiscus, in one embodiment. The present disclosure, in one embodiment, is a method of enhancing OPH activity comprising feeding a human or non-human organism an extract of one or more plants selected from the group consisting of fenugreek, fenugreek and hibiscus. Regarding. The OPH activity enhancing method of the present disclosure, in one or more embodiments, replaces the above extract with the OPH activity enhancing agent, beverage composition, food composition, functional food, cosmetics, supplement, quasi-drug of the present disclosure. Foreign products or medicines may be used.

[Manufacturing method of beverage composition, food composition, functional food, cosmetics, supplement, quasi-drug or pharmaceutical product for enhancing OPH activity]
The present disclosure relates to, in other aspects, a method for producing a beverage composition, a food composition, a functional food, a cosmetic, a supplement, a quasi-drug, or a pharmaceutical product (a method for producing the beverage composition, etc. of the present disclosure). The method for producing a beverage composition or the like of the present disclosure is an extract of fenugreek in a beverage composition, a food composition, a functional food, a cosmetic, a supplement, a quasi-drug, or a pharmaceutical product in order to impart an OPH activity enhancing effect. Includes the addition of. The method for producing a beverage composition or the like of the present disclosure includes fenugreek, phenel and pharmaceutical products in a beverage composition, a food composition, a functional food, a cosmetic, a supplement, a quasi-drug or a pharmaceutical product in order to impart an OPH activity enhancing effect. It involves adding one or more plant extracts selected from the group consisting of hibiscus.

Examples of the extract include a water extract and an organic solvent extract of the plant in one or more embodiments. Examples of the water extract include hot water extracts and the like in one or more embodiments. The form of the extract may be an aqueous solution, a concentrated solution, a dried product or the like in one or more embodiments.

[Extract composition for enhancing OPH activity]
The present disclosure relates to, in other embodiments, an extraction composition of one or more plants selected from the group consisting of fenugreek, phenel and hibiscus for enhancing oxidative proteolytic enzyme activity. In the present disclosure, the "extracted composition" refers to a liquid or solid obtained by extracting a plant. Examples of the extraction composition include, in one or more embodiments, a supernatant and a filtrate obtained by solvent-extracting a plant. Examples of the extraction solvent include water, alcohols such as methanol and ethanol, ether and the like in one or more embodiments. In one or more embodiments, the form of the extract composition includes an aqueous solution, a concentrated solution, a dried product, and the like. The extract compositions of the present disclosure can be used as food ingredients in one or more embodiments. In the present disclosure, the "food raw material" means a material or raw material used for manufacturing foods such as processed foods and health foods and supplements. Examples of the form of the food raw material include an aqueous solution, a concentrated solution, a dried product, and the like in one or a plurality of embodiments.

The present disclosure may relate to one or more of the following embodiments:
[A1] An oxidative proteolytic enzyme activity enhancer containing an extract of fenugreek as an active ingredient.
[A2] The oxidative proteolytic enzyme activity enhancer according to [A1], which further comprises an extract of at least one of phenel and hibiscus.
[A3] An oxidizing proteolytic enzyme activity enhancer containing an extract of one or more kinds of plants selected from the group consisting of fenugreek, fenugreek and hibiscus as an active ingredient.
[A4] The oxidative proteolytic enzyme activity enhancer according to any one of [A1] to [A3], which comprises an extract of fenugreek, fenugreek and hibiscus as an active ingredient.
[A5] The oxidative proteolytic enzyme activity enhancer according to any one of [A1] to [A4], wherein the extract is at least one of a water extract and an organic solvent extract.
[A6] The oxidative proteolytic enzyme activity enhancer according to any one of [A1] to [A5], which comprises a dried product of the extract as an active ingredient.
[A7] Beverage compositions, food compositions, functional foods, cosmetics, supplements, quasi-drugs or products containing the oxidative proteolytic enzyme activity enhancer according to any one of [A1] to [A6] as an active ingredient. Pharmaceuticals.
[A8] A beverage composition, a food composition, for enhancing the activity of oxidative proteolytic enzyme, which comprises an extract of one or more kinds of plants selected from the group consisting of fenugreek, fenugreek and hibiscus as an active ingredient. Functional foods, cosmetics, supplements, quasi-drugs or pharmaceuticals.
[A9] A method for producing an oxidative proteolytic enzyme activity enhancer, which comprises extracting fenugreek with a solvent.
[A10] In order to impart the effect of enhancing the activity of oxidative proteolytic enzyme, a beverage composition, a food composition, a functional food, a cosmetic, a supplement, a quasi-drug or a pharmaceutical product is selected from the group consisting of phenuglique, phenel and hibiscus. A method for producing a beverage composition, a food composition, a functional food, a cosmetic, a supplement, a quasi-drug, or a pharmaceutical, which comprises adding one or more kinds of plant extracts.

[Glycation stress inhibitor]
In the present disclosure, a mixture of water extracts of Fenuglique (scientific name: Trigonella foenum-glaecum) and Fennell (scientific name: Foenicularum vulgare), particularly hot water extracts, is caused by oxidative stress and glycation stress in vivo such as oxidative protein and glycated protein. It has a new action of activating an oxidative protein-degrading enzyme (OPH), which is known to preferentially decompose an AGEs-degrading agent, and has a higher AGEs resolution than PTB known as an AGEs-degrading agent. Based on knowledge.
The present disclosure is based on the new finding that fenugreek water extracts, especially hot water extracts, have amylase and α-glucosidase inhibitory activity.
The present disclosure is also based on the new finding that the use of fenugreek extract and fenugreek extract in combination can reduce aberrant proteins in one or more embodiments.
The present disclosure is based on the new finding that glycation stress can be multifacetedly suppressed in one or more embodiments by using a fenugreek extract and a fenugreek extract in combination.

Glycation stress is considered to be a concept that comprehensively captures the stress on the living body due to the loading of reducing sugars and aldehydes and the subsequent reaction. In the present disclosure, "suppressing glycation stress" includes suppressing the production of abnormal protein, degrading and / or removing the abnormal protein, and suppressing hyperglycemia. In addition, suppressing glycation stress may include, in one or more embodiments, suppressing the production of an oxidizing substance that causes abnormal protein production, or decomposing or eliminating the oxidizing substance.

"Abnormal protein" means a protein that has undergone oxidation, saccharification, aldehyde modification, or the like. The abnormal protein may be a product of a Maillard reaction (Maillard reaction product) in one or more embodiments. Maillard reaction products include, in one or more embodiments, intermediates produced by the Maillard reaction and end products produced by the Maillard reaction. Examples of the Maillard reaction product include polypeptides and proteins containing AGEs, polypeptides and proteins having a crosslinked structure by AGEs, and glycated proteins produced by the Maillard reaction in one or more embodiments. Abnormal proteins include, in one or more embodiments, polypeptides and proteins with Schiff bases / Schiff bases formed, Amadori compounds (ketoamines, glycated proteins), polypeptides and proteins with α-dicarbonyl compounds, and Examples thereof include polypeptides and proteins having AGEs.

Suppressing the production of an abnormal protein includes, in one or more embodiments, suppressing the production of a substance that is directly or indirectly involved in the production of the abnormal protein. Suppressing the production of the substance includes, in one or more embodiments, amylase inhibition, α-glucosidase inhibition and the like. Substances directly or indirectly involved in the production of abnormal proteins include, in one or more embodiments, α-dicarbonyl compounds such as reducing sugars, glyoxal, methylglyoxal and glyceraldehyde, compounds having a shift base, and amadori compounds. , Aldehydes, AGEs and the like. Examples of AGEs include amino acids and dipeptides saccharified by the Maillard reaction in one or more embodiments, and examples include, but are not limited to, pentosidine, crosline, pyropyridine, pyrarinine, carboxymethyllysine (CML), and carboxyethyllysine. , Carboxymethylarginine (CMA), argpyrimidine, imidazolone compounds, glyoxal-lysine dimer (GOLD), methyl glyoxal-lysine dimer (MOLD) and the like.

Degradation and / or removal of anomalous proteins includes, in one or more embodiments, degradation of the Maillard reaction product. Degrading the Maillard reaction product comprises, in one or more embodiments, cleaving the crosslinked structure between the polypeptide and / or protein by AGEs formed with the Maillard reaction. Degrading the Maillard reaction product includes, in one or more embodiments, other polypeptides and proteins containing AGEs, polypeptides and proteins having a crosslinked structure with AGEs, or glycated proteins produced by the Maillard reaction. May include disassembling. Further, in one or a plurality of embodiments without particular limitation, CC of a diketone structure contained in a polypeptide and protein containing AGEs, a polypeptide and protein having a crosslinked structure by AGEs, or a glycated protein produced by the Maillard reaction. It may include breaking the bond.

Suppressing the rapid rise in blood glucose level includes, in one or more embodiments, inhibiting amylase, inhibiting α-glucosidase, inhibiting the production of reducing sugars, and the like. Can be mentioned. Examples of the reducing sugar include monosaccharides such as glucose and fructose, maltose-type disaccharides and oligosaccharides such as lactose and maltose in one or more embodiments.

Suppressing hyperglycemia may include, in one or more embodiments, suppressing a rapid rise in blood glucose levels.

Examples of suppressing oxidative stress include the use of a substance having a component having high antioxidant power.

The present disclosure relates to a glycation stress inhibitor (the glycation stress inhibitor of the present disclosure) containing fenugreek and fenugreek extracts as active ingredients in one embodiment.

The glycation stress inhibitor of the present disclosure refers to an agent having at least Maillard reaction product decomposing activity (also referred to as AGEs degrading activity) and OPH activity enhancing ability. The saccharification stress inhibitor of the present disclosure may have one or more functions such as amylase inhibitory action, α-glucosidase inhibitory action, and antioxidant activity as additional functions in one or more embodiments. In addition, the saccharification stress inhibitor of the present disclosure suppresses, decomposes or removes, and decomposes, removes or suppresses the production of a substance that causes abnormal protein production in one or more embodiments. It can be carried out.

The saccharification stress inhibitor of the present disclosure contains fenugreek and an extract of fenugreek as active ingredients. Therefore, according to the saccharification stress inhibitor of the present disclosure, in one or more embodiments, the decomposition efficiency of the oxidized protein and the saccharified protein can be improved by enhancing the enzymatic activity of OPH, performing cross-linking cleavage, and the like. sell. According to the glycation stress inhibitor of the present disclosure, in one or more embodiments, the accumulation of AGEs is promoted by promoting the decomposition of the cross-linked structure between the polypeptide and / or the protein caused by the AGEs produced in vivo. Can be suppressed. According to the glycation stress inhibitor of the present disclosure, since it has an amylase or α-glucosidase inhibitory action in one or more embodiments, it suppresses a rapid increase in blood glucose level and is non-enzymatic of reducing sugar and protein. It is possible to suppress the reaction failure of the TCA circuit due to the presence of excessive glucose and the like, and as a result, the production of abnormal protein can be suppressed. Further, since the glycation stress inhibitor of the present disclosure has an antioxidant effect, it is possible to suppress the production and accumulation of various abnormal proteins related to glycation stress and oxidative stress from various aspects. Therefore, according to the saccharification stress inhibitor of the present disclosure, saccharification stress and / or oxidative stress can be suppressed or reduced in a multifaceted manner in one or more embodiments. Further, according to the glycation stress inhibitor of the present disclosure, anti-aging, anti-aging effect or preventive effect of lifestyle-related diseases can be expected in one or more embodiments. Therefore, the glycation stress inhibitor of the present disclosure can also be referred to as an antioxidant stress agent, an anti-aging agent, an antiaging agent or a lifestyle-related disease preventive agent in one or more embodiments.

Since the saccharification stress inhibitor of the present disclosure uses plant-derived extracts such as fenugreek and fennel as active ingredients, it can have the effects of being excellent in safety and productivity.

In the glycation stress inhibitor of the present disclosure, the fenugreek extract and the fenugreek extract may be blended in the same ratio (1 part by weight of the fenugreek extract to 1 part by weight of the fenugreek extract). , May be blended in different proportions.

The blending ratio (dry weight ratio) of the fenugreek extract and the fenugreek extract in the saccharification stress inhibitor of the present disclosure is, in one or more embodiments, the fenugreek extract per part by weight of the fenugreek extract. The amount is 0.1 to 10 parts by weight, and the extract of fenugreek is 0.2 to 5 parts by weight from the viewpoint of further improving the saccharification stress suppressing effect.

The saccharification stress inhibitor of the present disclosure may contain an extract of hibiscus (scientific name: Hibiscus sabdariffa) as a further active ingredient from the viewpoint of further improving the antioxidant power and the amylase inhibitory action in one or more embodiments. .. The saccharification stress inhibitor of the present disclosure comprises, in one or more embodiments, an extract of fenugreek, phenel and hibiscus as an active ingredient.

When the saccharification stress inhibitor of the present disclosure further comprises an extract of hibiscus, the extract of hibiscus is in the same proportion as the extract of fenugreek and / or the extract of fenugreek (1 weight of extract of fenugreek or extract of fenugreek). It may be blended in 1 part by weight with respect to the portion), or may be blended in different ratios.

The saccharification stress suppressants of the present disclosure include, in one or more embodiments, an extract of fenugreek, an extract of fenugreek and an extract of hibiscus. In this embodiment, the blending ratio (dry weight ratio) of the phenuglique extract, the phenell extract and the hibiscus extract is 0.05 to 20 parts by weight of the phenus extract per 1 part by weight of the phenuglique extract. , The hibiscus extract is 0.05 to 20 parts by weight, and the phenel extract is 0.1 to 10 parts by weight, and the hibiscus extract is 0 from the viewpoint of further improving the antioxidant power and the amylase inhibitory action. .1 to 10 parts by weight.

The glycation stress inhibitor of the present disclosure contains at least one of lemon balm (scientific name: Melissa officeis) and rosemary (scientific name: Rosmarinus officeis) as further active ingredients from the viewpoint of further improving the resolution of AGEs in one or more embodiments. It may be included.

The extract may be extracted from any part. Examples of the site of use include whole plants, flowers, leaves, seeds, fruits, branches, stems, roots, calyxes, bracts and the like in one or more embodiments. The site to be used may be one type, or two or more types of sites may be used in combination. Phenels are preferably seeds in one or more embodiments. Fenugreek is preferably seeds in one or more embodiments. The hibiscus is preferably a shoeblackplant and a bract in one or more embodiments. Rosemary is preferably leaves in one or more embodiments. Lemon balm is preferably leaves in one or more embodiments.

The extraction method is not particularly limited, and examples thereof include solvent extraction and the like in one or more embodiments. Examples of the extraction solvent include water, alcohols such as methanol and ethanol, ether and the like in one or more embodiments. Examples of the extract include a water extract, an organic solvent extract, and the like in one or more embodiments. Examples of the water extract include hot water extracts and the like in one or more embodiments. The temperature of the solvent extraction may be 60 to 100 ° C., 70 to 90 ° C., 80 ° C. or the like in one or more embodiments. In one or more embodiments, the solvent extraction time may be 30 minutes to 5 hours, 1 to 5 hours, 2 to 5 hours, 3 to 5 hours, 1 hour, or the like. The temperature of hot water extraction may be 60 to 100 ° C., 70 to 90 ° C., 80 ° C. or the like in one or more embodiments. In one or more embodiments, the hot water extraction time may be 30 minutes to 5 hours, 1 to 5 hours, 2 to 5 hours, 3 to 5 hours, 1 hour, or the like. The form of the extract may be an aqueous solution, a concentrated solution, a dried product or the like in one or more embodiments. Drying can be performed in one or more embodiments by heat-treating the extract obtained by solvent extraction. The heating temperature is 70-150 ° C or 110 ° C in one or more embodiments. The heating time is 1-10 hours or 4 hours in one or more embodiments.
Suitable methods for producing the saccharification stress inhibitor of the present disclosure are as follows in one or more embodiments. An appropriate amount of phenuglique powder (eg, 2 g) and fennel powder (eg, 2 g) are extracted with an appropriate amount of distilled water (eg, 40 mL) at 80 ° C. for 1 hour, then cooled to room temperature, and then the obtained slurry is obtained. After filtration, the resulting filtrate is used as a plant extract. When using dried plant extracts, the resulting plant extracts are placed on aluminum trays and dried, for example, in an incubator set at 110 ° C. for 4 hours.

The form of the saccharification stress inhibitor of the present disclosure is not particularly limited, and examples thereof include solids, granules, powders, pastes, and liquids in one or more embodiments.

The saccharification stress inhibitor of the present disclosure may contain other extracts and other components in one or more embodiments as long as the active ingredient does not inhibit the saccharification stress suppression function.

The saccharification stress inhibitor of the present disclosure can be used as a raw material for beverage compositions, food compositions, functional foods, cosmetics, supplements, feeds, quasi-drugs, pharmaceuticals and the like in one or more embodiments. The form as a raw material is not particularly limited, and examples thereof include solids, granules, powders, pastes, and liquids in one or more embodiments.
The present disclosure relates, as one or more embodiments, to beverage compositions, food compositions, functional foods, cosmetics, supplements, feeds, quasi-drugs or pharmaceuticals comprising the saccharification stress suppressors of the present disclosure.

The daily intake of the glycation stress inhibitor of the present disclosure is not particularly limited, and is 5 mg or more or 2000 mg or less in one or more embodiments.

[Manufacturing method of glycation stress inhibitor]
The present disclosure relates to, in other embodiments, a method for producing a glycation stress inhibitor, which comprises solvent extraction of fenugreek and fenugreek. According to the production method of the present disclosure, the saccharification stress inhibitor of the present disclosure can be produced.

The production method of the present disclosure may include, in one or more embodiments, mixing an extract of fenugreek with an extract of fenugreek to obtain a mixture of the extracts. The mixing ratio may be the same ratio (1 part by weight of fenugreek extract to 1 part by weight of fenugreek extract) or different ratios in one or more embodiments.

The production method of the present disclosure comprises mixing these extracts in one or more embodiments so that the extract of fenugreek is 0.1-10 parts by weight per part by weight of the extract of fenugreek. However, from the viewpoint of producing a saccharification stress inhibitor capable of further improving the saccharification stress suppressing effect, it may be included to mix these extracts so that the extract of fenugreek is 0.2 to 5 parts by weight. good.

The production method of the present disclosure may include, in one or more embodiments, mixing fenugreek and fenugreek and solvent-extracting the resulting mixture to obtain a mixed extract. The mixing ratio may be the same ratio (1 part by weight of fenugreek to 1 part by weight of fenugreek) or different ratios in one or more embodiments. Further, the mixture may be mixed so that the ratio of the extract of each plant in the obtained mixed extract is the same ratio (1 part by weight of the extract of fenugreek to 1 part by weight of the extract of fenugreek), or different. It may be mixed so as to be a ratio.

The method for producing a glycation stress inhibitor of the present disclosure further comprises, in one or more embodiments, solvent extraction of at least one plant selected from the group consisting of hibiscus, lemon balm, and rosemary. good.

The production method of the present disclosure may include, in one or more embodiments, solvent extraction of a dry plant.

The production method of the present disclosure may include, in one or more embodiments, filtering or centrifuging the extract obtained by solvent extraction.

The production method of the present disclosure may comprise, in one or more embodiments, drying the resulting extract.

The production method of the present disclosure comprises, in one or more embodiments, mixing an extract of fenugreek, an extract of fenugreek, and an extract of a plant other than these to obtain a mixture of extracts. May be good. The mixing ratio is, in one or more embodiments, the same ratio as the fenugreek extract and / or the fenugreek extract (1 part by weight with respect to 1 part by weight of the fenugreek extract or fenugreek extract). It may be a different ratio.

In one or more embodiments, the production method of the present disclosure comprises 0.05 to 20 parts by weight of the extract of amylase and 0.05 to 20 parts by weight of the extract of hibiscus per 1 part by weight of the extract of phenuglique. From the viewpoint of producing a saccharification stress inhibitor capable of further improving the antioxidant power and the amylase inhibitory effect, the Fennell extract may be 0.1 to 10 weight by weight. Parts may include mixing these extracts so that the hibiscus extract is 0.1 to 10 parts by weight.

The production method of the present disclosure may include, in one or more embodiments, mixing fenugreek, fennel, and plants other than these, and solvent-extracting the resulting mixture to obtain a mixed extract. good. The mixing ratio may be the same ratio of fenugreek and / or fenugreek (1 part by weight to 1 part by weight of fenugreek or fenugreek) or different ratios in one or more embodiments. Further, the mixture may be mixed so that the ratio of the extract of each plant in the obtained mixed extract is the same (1 part by weight with respect to 1 part by weight of the extract of fenugreek or the extract of fenugreek). They may be mixed in different proportions.

[Method of suppressing glycation stress]
The present disclosure relates to, in one embodiment, a method for suppressing glycation stress, which comprises administering an extract of fenugreek and fenugreek to a living body. The saccharification stress suppression method of the present disclosure comprises, in one or more embodiments, further administering to the living body an extract of at least one plant selected from the group consisting of hibiscus, lemon balm, and rosemary. May be good. The saccharification stress suppression method of the present disclosure, in one or more embodiments, replaces the above extract with the saccharification stress inhibitor, beverage composition, food composition, functional food, cosmetics, supplement, quasi-drug of the present disclosure. Foreign products or medicines may be used.

[Beverage composition, food composition, functional food, cosmetics, supplement, quasi-drug or pharmaceutical manufacturing method for imparting glycation stress suppressing function]
The present disclosure relates to, in other aspects, a method for producing a beverage composition, a food composition, a functional food, a cosmetic, a supplement, a quasi-drug, or a pharmaceutical product (a method for producing a second beverage composition, etc. of the present disclosure). .. The second method for producing a beverage composition, etc. of the present disclosure is to add fenugreek to a beverage composition, a food composition, a functional food, a cosmetic, a supplement, a quasi-drug, or a pharmaceutical product in order to impart a saccharification stress suppressing effect. And adding an extract of fenugreek.

The method for producing the second beverage composition and the like of the present disclosure further comprises, in one or more embodiments, an extract of at least one plant selected from the group consisting of hibiscus, lemon balm, and rosemary. It may include additions to products, food compositions, functional foods, cosmetics, supplements, quasi-drugs or pharmaceuticals.

Examples of the extract include a water extract and an organic solvent extract of the plant in one or more embodiments. Examples of the water extract include hot water extracts and the like in one or more embodiments. The form of the extract may be an aqueous solution, a concentrated solution, a dried product or the like in one or more embodiments.

[Extract composition for imparting glycation stress suppressing function]
The present disclosure relates to other embodiments relating to fenugreek and fenugreek extract compositions for imparting a glycation stress-suppressing function. Examples of the extraction composition include, in one or more embodiments, a supernatant and a filtrate obtained by solvent-extracting a plant. Examples of the extraction solvent include water, alcohols such as methanol and ethanol, ether and the like in one or more embodiments. In one or more embodiments, the form of the extract composition includes an aqueous solution, a concentrated solution, a dried product, and the like. The extract compositions of the present disclosure can be used as food ingredients in one or more embodiments. Examples of the form of the food raw material include an aqueous solution, a concentrated solution, a dried product, and the like in one or a plurality of embodiments.

The extract composition of the present disclosure may comprise, in one or more embodiments, an extract of at least one plant selected from the group consisting of hibiscus, lemon balm, and rosemary.

The present disclosure may relate to one or more of the following embodiments:
[B1] A glycation stress inhibitor containing fenugreek and fenugreek extracts as active ingredients.
[B2] The saccharification stress inhibitor according to [B1], which further comprises an extract of hibiscus as an active ingredient.
[B3] The saccharification stress inhibitor according to [B1] or [B2], wherein the extract is at least one of a water extract and an organic solvent extract.
[B4] The saccharification stress inhibitor according to any one of [B1] to [B3], which comprises a dried product of the extract as an active ingredient.
[B5] A beverage composition, a food composition, a functional food, a cosmetic, a supplement, a quasi-drug, or a pharmaceutical product containing the saccharification stress inhibitor according to any one of [B1] to [B4] as an active ingredient.
[B6] Beverage compositions, food compositions, functional foods, cosmetics, supplements, quasi-drugs or pharmaceuticals for suppressing saccharification stress containing fenugreek and fenugreek extracts as active ingredients.
[B7] A method for producing a glycation stress inhibitor, which comprises extracting fenugreek and fenugreek with a solvent.
[B8] The present invention comprises adding an extract of phenuglique and phenell to a beverage composition, a food composition, a functional food, a cosmetic, a supplement, a non-pharmaceutical product or a pharmaceutical product in order to impart a saccharification stress suppressing function. Beverage composition, food composition, functional food, cosmetics, supplements, non-pharmaceutical products or methods for manufacturing pharmaceutical products.

Hereinafter, the present disclosure will be further described with reference to examples. However, this disclosure is not construed as being limited to the following examples.

[Preparation of plant extract]
2 g of the dry powder of the plant shown in Table 1 below was extracted with 40 ml of distilled water at 80 ° C. for 1 hour, cooled to room temperature, filtered, and the obtained filtrate was used as a plant extract.
The solid content concentration of each sample was calculated by placing 5 mL of the obtained filtrate in an aluminum tray and drying it in an incubator set at 110 ° C. for 4 hours, and then measuring the residual amount. The results are shown in Table 1 below.

[OPH activation confirmation test (1)]
As OPH, Acylamino-acid releasing enzyme (AARE) (manufactured by Takara Bio Inc.) was used, dissolved in the attached sodium phosphate buffer to adjust to 0.5 U / ml, and then 0.2 mol / L Tris-. It was used after being diluted 500-fold with HCl (pH 7.4). As a reaction substrate for OPH, a 0.05 mol / L N-actyl-L-alanine p-nitroaniline (AAPA) 50% ethanol solution was used.
The plant extract, OPH solution, AAPA solution and Tris-HCl of Table 1 below were added to a 96-well microplate (PerkinElmer) so as to have the blending amounts shown in Table 2 below. The microplate was reacted in a microplate reader (SpectraMax Paradigm Multi-Mode Microplate Reader) (manufactured by Molecular Device) whose temperature was adjusted to 37 ° C. for 48 hours (2880 minutes), and the absorbance at a wavelength of 405 nm was measured over time.

OPH activity was assessed by the results of the 24-hour reaction. OPH activation rate of each plant extract from the following formula using the absorbance of each reaction solution A, B, C and D as a relative value with the decomposition activity of AAPA by OPH (without addition of plant extract) as 100% ( %) Was calculated. The results are shown in Table 1 below as relative values with the activation rate of lemon balm as 100.
OPH activation rate (%) = (AB) / (CD) × 100

As shown in Table 1, fenugreek, hibiscus and fenugreek showed a 5-fold or more OPH activity-increasing effect as compared with lemon balm.

[Preparation of plant extract powder]
Each plant in Table 3 below is extracted with 15 times the amount of water and at a temperature of 100 ° C for 30 minutes, solid-liquid separated and concentrated, and then sterilized within 100 ° C or more and within 30 minutes is spray-dried to obtain a plant-extracted powder. did.

[Preparation of sample solution]
Each of the above plant extract powders was diluted with distilled water to a concentration of 5 mg / ml to prepare a sample solution.

[OPH activation confirmation test (2)]
Acylamino-acid releasing enzyme (AARE) (manufactured by Takara Bio Inc.) is dissolved in the attached sodium phosphate buffer solution, and 0.2 mol / L Tris-HCl (pH 7.4) is used at 0.01 U / ml, 0.005 U. / Ml or 0.001 U / ml OPH solution was prepared. As a reaction substrate for OPH, a 0.05 mol / L N-actyl-L-alanine p-nitroaniline (AAPA) 50% ethanol solution was used.
OPH solution, AAPA solution, and sample solution (5 mg / ml) were mixed and added to each well of the 96-well microplate, and the mixture was reacted in an incubator set at 37 ° C. for 24 hours. The absorbance of the reaction solution at 405 nm was measured with a microplate reader.

The enzyme activity of OPH was determined as the amount of change in absorbance (reaction rate) per hour (OPH reaction rate of the sample). At the same time, the same measurement was performed without adding the sample solution as a reference (Ref), and the reaction rate was determined (OPH reaction rate of Ref). The OPH activation rate (%) was calculated from the following formula using the OPH reaction rate of the sample and the OPH reaction rate of Ref (n = 3). The results are shown in Table 3 below.
OPH activation rate (%) = [(OPH reaction rate of sample) / (OPH reaction rate of Ref)] × 100

Epigallocatechin gallate (EGCg) (5 mg / ml) was used as a negative control and similar measurements were performed. The results are shown in Table 3 below.

As shown in Table 3, it was confirmed that the OPH activity increasing effect exceeded 170% in each case. In addition, even when fenugreek, hibiscus and fenugreek were used in combination, it was confirmed that the OPH activity increasing effect was equal to or higher than that of fenugreek alone.

[AGEs Crosslink Cutting Action Confirmation Test (Part 1)]
The procedure was performed according to the description of Sara Vasan et al. Nature, 382, pp275-278 (1996).
As a substrate, 1-Phenyl-1,2-propanedione (PPD), which is a reaction substrate of an AGEs cross-linking model having an α-diketone structure, was used, and as a positive control, N-phenacylthiazolium bromide (PTB) was used.
The plant extract, 10 mmol / L PPD, and 0.2 mol / L phosphate buffer (pH 7.4) were mixed at a ratio of 5: 1: 4 and reacted at 37 ° C. for 8 hours (n = 3). After completion of the reaction, hydrochloric acid was added to terminate the reaction to obtain a reaction solution.
The reaction mixture was centrifuged at 3,000 × g at 20 ° C. for 10 minutes, and the amount of benzoic acid in the supernatant was analyzed by reverse phase HPLC. The amount of benzoic acid in the reaction solution was determined by subtracting the amount of benzoic acid in the separately measured plant extract. Since 1 mol of PPD produces 1 mol of benzoic acid, the cross-linking cleavage rate was calculated from the following formula. The results are shown in Table 4 below.
Crosslink cutting rate (%) = {(AB) / C} × 100
A: Amount of benzoic acid in the reaction solution B: Amount of benzoic acid in the plant extract C: Amount of PPD subjected to the reaction (base mass)

As a control, 10 mmol / L PTB was used instead of the plant extract.

As shown in Table 4, all of Fennell, Lemon Balm and Rosemary showed higher cross-linking cleavage rates than PTB reported as a compound that degrades cross-linking structures involving AGEs. In particular, Fennell showed a cross-linking cleavage rate nearly twice that of PTB.

[AGEs Crosslink Cutting Action Confirmation Test (Part 2)]
The procedure was performed according to the description of Sara Vasan et al. Nature, 382, pp275-278 (1996).
SuperBlock T20 (PBS) Blocking Buffer was added to a commercially available 96-well microplate coated with collagen at 300 μL / well and reacted at 37 ° C. for 1 hour. After the reaction, the cells were washed with 0.05% Polyethylene (20) sorbitan monolaurate (Tween 20) / PBS (-), and then pre-AGEs bovine serum albumin (AGE-BSA) was added to each well of the microplate at 37 ° C. The cross-linking reaction was carried out for 4 hours. After the reaction, the cells were washed with Tween 20 / PBS (-), and then a plant extract was added to each well, and the cleavage reaction was carried out at 37 ° C. for 18 hours. Then, it was washed with Tween 20 / PBS (-), a primary antibody (anti-albumin bovine serum rabbit-polyclonal) was added, and the mixture was reacted at room temperature for 30 minutes. Furthermore, after washing with Tween 20 / PBS (-), a secondary antibody (goat anti-rabbit IgG horseradish peroxidase (HRP) conjugate) was added and reacted at room temperature for 30 minutes. After that, it was washed with Tween 20 / PBS (-), and TMB One Component HRP Microwell Substrate was added. Further, a PEG6000 solution was added and the reaction was carried out for 20 minutes. 1N HCl was added as a reaction stop solution, and the absorbance at 450 nm (main wavelength) / 630 nm (secondary wavelength) was measured with a microplate reader. The residual amount of AGE-BSA was determined from the calibration curve in which the amount of AGE-BSA was changed. The AGE-BSA decomposition rate (collagen crosslink decomposition rate) was determined using the following formula. The results are shown in Table 5 below.
AGE-BSAs decomposition rate = {(added AGE-BSA amount-residual AGE-BSA amount) / added AGE-BSA amount} x 100

As a control, a similar test was conducted using 5 mmol / L punicalagin, which is known to have a Maillard reaction activity inhibitory effect, instead of the plant extract.

As shown in Table 5, it was confirmed that fenugreek, lemon balm and fenugreek had collagen cross-linking degrading activity. In particular, it was confirmed that lemon balm has collagen cross-linking decomposition activity equivalent to that of punicalagin.

[AGEs Crosslink Cutting Action Confirmation Test (Part 3)]
The same measurement as in the AGEs cross-linking cleavage action confirmation test (No. 1) was performed except that the mixed solution of the plant extracts shown in Table 6 below was used. The results are shown in Table 6 below.

As shown in Table 6, the mixed solution of fenugreek, hibiscus and fenugreek showed an AGEs cross-linking cleavage rate nearly twice that of PTB.

[Amylase inhibitory effect confirmation test]
The Enzy Chrom α-Amylase Assay Kit (BioAssay System) and α-Amylase from Porcine pancreas (Sigma) were used.
10 μL of the enzyme, Assay Buffer, and Glucose Standard included in the kit was dispensed into each well of the microplate. Then, 40 μL of the reaction solution containing the plant extract shown in Table 7 below was dispensed into each well, and the mixture was reacted for 15 minutes after stirring. Then, 150 μL of the color-developing reagent was dispensed into each well, reacted at room temperature for 20 minutes, and then the absorbance at 585 nm was measured to determine the inhibition rate and IC50 (n = 3). The obtained IC50s are shown in Table 7 below.

As shown in Table 7, the IC50s of fenugreek and hibiscus were both less than 0.1 mg / mL, and it was confirmed that fenugreek and hibiscus had high inhibitory activity on amylase. This suggests that fenugreek and hibiscus can suppress the rapid rise in blood glucose level, and as a result, can suppress the production of dicarbonyl compounds and AGEs that can cause the production of abnormal proteins.

[Α-Glucosidase inhibitory effect confirmation test]
QuantiChrom α-Glucosidase Assay Kit DAGD-100 (Bio Assay Sytem) and rat small intestinal acetone powder (Sigma, α-Glucosidase) were used.
After setting the temperature of the measuring part of the microplate reader to 30 ° C, 200 μL of the sample solution (including 8 uL of the substrate solution) containing the plant extract shown in Table 8 below and 20 μL of the α-glucosidase solution were added to each well of the microplate. The reaction was started by dispensing and stirring, and the change in absorbance at 405 nm was measured for 30 minutes. As a positive control of the inhibitory action, the α-glucosidase inhibitor acarbose was used, and the α-glucosidase activity inhibition rate (%) was calculated according to the following formula (n = 3), and IC50 was determined.
α-Glucosidase activity inhibition rate (%) = {1- (A / B)} × 100
A: Value of each concentration of sample and positive control (ΔAbs 30 min)
B: TOTAL (total color development) value (ΔAbs 30 min)

As shown in Table 8, it was confirmed that fenugreek and fenugreek have α-glucosidase inhibitory activity. This suggests that fenugreek and fenugreek can suppress a rapid increase in blood glucose level, and as a result, can suppress the production of dicarbonyl compounds and AGEs that can cause the production of abnormal proteins.

[Antioxidant confirmation test]
Arkray's Spotchem ™ i-Pack Oxystress Test ™ was used.
90 μL of the spotchem reagent r1 containing thiocyanate and 36 μL of the spotchem reagent r2 containing iron (iron) chloride III were mixed. To 126 μL of this mixed solution, 18 μL of the plant extract shown in Table 9 below was added, and the absorbance at 465 nm was measured using a spot chem to determine the antioxidant power (μmol / L).
On the other hand, the standard reagent ascorbic acid was used instead of the plant extract, and the absorbance was measured in the same manner to prepare a calibration curve showing the correlation between the ascorbic acid concentration and the absorbance. Then, the antioxidant power was converted into ascorbic acid equivalent (mg / L) from the absorbance of the plant extract and the calibration curve. The results are shown in Table 9 below.

As shown in Table 9, it was confirmed that hibiscus has high antioxidant power. This suggests that hibiscus can eliminate the oxidizing substances that cause abnormal protein production.

Claims (13)

An oxidative proteolytic enzyme activity enhancer containing a dried product of a water extract of fenugreek seeds and a dried product of a water extract of fenugreek seeds as active ingredients. The oxidative proteolytic enzyme activity enhancer according to claim 1, further comprising one or two selected from the group consisting of a dried product of a water extract of hibiscus buds and a dried product of a water extract of hibiscus bracts. .. The oxidative proteolytic enzyme activity enhancer according to claim 1 or 2, wherein the water extract is a hot water extract. A saccharification stress inhibitor containing a dried product of a water extract of fenugreek seeds and a dried product of a water extract of fenugreek seeds as active ingredients. The saccharification stress inhibitor according to claim 4, further comprising one or two selected from the group consisting of a dried water extract of hibiscus bracts and a dried water extract of hibiscus bracts. The saccharification stress inhibitor according to claim 4 or 5, wherein the water extract is a hot water extract. Enhancement of oxidative proteolytic enzyme activity and saccharification stress using the oxidative proteolytic enzyme activity enhancer according to any one of claims 1 to 3 or the saccharification stress inhibitor according to any one of claims 4 to 6 as an active ingredient. Beverage compositions, food compositions, functional foods, cosmetics, supplements, or quasi-drugs for at least one of the suppressions . A beverage containing, as an active ingredient, a dried product of a water extract of fenugreek seeds and a dried product of a water extract of fenugreek seeds for enhancing at least one of oxidative proteolytic enzyme activity and suppressing saccharification stress. Compositions, food compositions, functional foods, cosmetics, supplements, or quasi-drugs. The beverage composition according to claim 8, further comprising one or two dried products selected from the group consisting of a dried product of a water extract of hibiscus buds and a dried product of a water extract of hibiscus buds. Food composition, functional food, cosmetics, supplements, or quasi-drugs. A method for producing at least one of an oxidative proteolytic enzyme activity enhancer and a saccharification stress inhibitor, which comprises mixing a dried product of a water extract of fenugreek seeds and a dried product of a water extract of fenugreek seeds. 17. Production method. Water extraction of phenuglique seeds in beverage compositions, food compositions, functional foods, cosmetics, supplements, or non-pharmaceutical products to impart at least one of the oxidative proteolytic enzyme activity enhancing effect and the saccharification stress suppressing function. A method for producing a beverage composition, a food composition, a functional food, a cosmetic, a supplement, or a non-pharmaceutical product, which comprises adding a dried product and a dried product of a water extract of Fennell seeds. 22. Production method.