AGEs reducing agents, methods for reducing AGEs

Abstract

To provide a novel AGEs reducing agent that has effective AGEs reduction capabilities. [Solution] The present invention is an AGEs reducing agent containing a sulfur-containing amino acid. It is preferable that the AGEs reducing agent suppresses the formation of AGEs or promotes their decomposition. It is preferable that the sulfur-containing amino acid has a sulfoxide group, and among these, it is preferable that it is a compound represented by the following formula (I). JPEG2026167640000007.jpg41170 (In the formula, R1 and R2 are bonded to each other to form a ring, or R1 is an alkyl group having 1 to 4 carbon atoms or an alkenyl group having 2 to 3 carbon atoms, and R2 is a hydrogen atom.)

Description

Technical Field

[0001] The present invention relates to an AGEs reducing agent and a method for reducing AGEs.

Background Art

[0002] The reaction in which an amino group of an amino acid, peptide or protein reacts with a ketone or aldehyde, particularly a reducing sugar such as glucose, to produce a brown pigment is referred to as the Maillard reaction. The substance produced as the end product of this Maillard reaction is called Advanced Glycation End Products, hereinafter sometimes referred to as "AGEs". The Maillard reaction consists of an "early reaction" in which an amino group and glucose or the like react non-enzymatically to form a Schiff base, followed by Amadori rearrangement, a "middle-stage reaction" in which active intermediates having a dicarbonyl group such as glyoxal, methylglyoxal, and 3-DG (3-deoxyglucosone) are generated from the Schiff base or Amadori compound, and a "late-stage reaction" in which the active intermediates further non-enzymatically react with amino groups, and undergo repeated dehydration and condensation reactions to form AGEs.

[0003] In animals such as humans, AGEs are absorbed from the intestine via ingestion of foods and beverages containing AGEs, that is, via diet, and are also formed in the body. In the latter case, for example, AGEs are produced by the binding of proteins to sugars or intermediate metabolites of sugars that can no longer be metabolized in the living body due to excessive obesity or aging. AGEs in the blood and AGEs accumulated outside tissue cells promote the enhancement of inflammation and oxidative stress by transmitting signals into cells via RAGE (Receptor of AGEs). As a result, it is considered that various organs are damaged, and diabetes, dementia, arteriosclerosis and the like occur, and suppressing the production of AGEs is considered to be extremely important for the prevention of these diseases (Non-Patent Documents 1 and 2).

[0004] In particular, pentosidine, a type of AGE, is known to bind to collagen and is a substance that causes a decrease in skin elasticity (Patent Document 1). In addition, methylglyoxal-hydroimidazolon, another type of AGE, has been reported to be associated with symptoms and diseases associated with diabetes and aging (Patent Document 2). Therefore, it is considered desirable to reduce the amount of these substances in the blood and tissues. Methods to reduce AGEs such as pentosidine and methylglyoxal-hydroimidazolon in the blood and tissues include inhibiting the formation of AGEs in the body or promoting the breakdown of AGEs in the body.

[0005] Conventionally, black garlic extract has been reported to have an inhibitory effect on AGEs formation by inhibiting the formation of 3-DG, an intermediate in the formation of AGEs (Patent Document 3). Similarly, onions, which are also plants of the Allium genus, have been reported to have an inhibitory effect on the glycation of aminophospholipids (Patent Document 4). However, while Patent Document 3 mentions S-allyl cysteine, cycloalliin, etc., in the section on prior art, it only lists other components such as Normeconin and 5-Hydroxymethyl furfural as active ingredients for anti-glycation. Furthermore, Patent Document 4 deals with a phenomenon different from the present invention, which is aminophospholipid glycation, and does not describe what in onion extract causes the glycation inhibitory effect.

[0006] Onions are known to contain sulfur-containing amino acids such as isoalliin, cycloalliin, and methiin (Patent Document 5), and Patent Document 6 describes a method for increasing the sulfur-containing amino acids in processed onion products. However, Patent Documents 5 and 6 do not describe anti-glycation effects. [Prior art documents] [Patent Documents]

[0007] [Patent Document 1] Japanese Patent Publication No. 2024-155038 [Patent Document 2] Japanese Patent Publication No. 2017-96837 [Patent Document 3] Japanese Patent Publication No. 2019-041760 [Patent Document 4] Japanese Patent Publication No. 2004-115491 [Patent Document 5] Japanese Patent Publication No. 2018-150283 [Patent Document 6] International Publication No. 2013 / 137284 [Non-patent literature]

[0008] [Non-Patent Document 1] CRITICAL REVIEWS IN TOXICOLOGY 2024, VOL. 54, No.8, p485-617 [Non-Patent Document 2] Comprehensive Medicine 2020, Vol.19, No.1,11-20 [Overview of the Initiative] [Problems that the invention aims to solve]

[0009] The object of the present invention is to provide an AGEs reducing agent and an AGEs reducing method that have effective AGEs reducing capabilities. [Means for solving the problem]

[0010] As a result of diligent research, the inventors discovered that a specific component exhibits excellent AGEs reduction effects, and thus completed the present invention. The present invention is based on the above findings and provides an AGEs reducing agent containing sulfur-containing amino acids.

[0011] Furthermore, the present invention provides a method for reducing AGEs by applying sulfur-containing amino acids to a reaction system that produces AGEs or to the produced AGEs.

[0012] Furthermore, the present invention provides a AGEs-reducing agent comprising an onion extract obtained in the step of subjecting a heated onion to treatment with a γ-glutamyl bond-cleaving enzyme, and subjecting the resulting enzyme-treated product to ion exchange chromatography. The present invention provides a AGEs-reducing agent comprising an onion extract obtained in the step of subjecting the resulting enzyme-treated product to ion exchange chromatography. Effects of the Invention

[0013] According to the present invention, it is possible to provide an AGEs-reducing agent and an AGEs reduction method that can be safely ingested orally and exhibit an effective AGEs formation inhibitory action or AGEs decomposition promoting action. Modes for Carrying Out the Invention

[0014] Hereinafter, preferred embodiments of the present invention will be described. The AGEs-reducing agent that is one embodiment of the present invention (hereinafter also referred to as "the agent of the present embodiment") contains a sulfur-containing amino acid. The agent of the present embodiment (the first embodiment) contains a sulfur-containing amino acid as an active ingredient. As sulfur-containing amino acids, amino acids having a sulfoxide group, a thiol group, or a sulfone group are known, and in the present embodiment, an amino acid containing a sulfoxide group is preferred.

[0015] The sulfoxide group is usually represented by R-S(=O)-R', and the S-O bond may be polarized to form S + -O - The structure may be such as described above. The agent of the present embodiment may use one type of sulfur-containing amino acid, particularly an amino acid containing a sulfoxide group, or may use two or more types in combination.

[0016] In the agent of the present embodiment, it is preferred that the sulfur-containing amino acid is not constituting a peptide, but is contained as a single amino acid (an amino acid that is not peptide-bonded to other amino acids) because the effects of the present invention are high. As the sulfur-containing amino acid to be the active ingredient in the present embodiment, for example, a compound represented by formula (I) is suitable.

[0017] Chemical Formula (In the formula, R1 and R2 are bonded to each other to form a ring, or R1 is an alkyl group having 1 to 4 carbon atoms or an alkenyl group having 2 to 3 carbon atoms, and R2 is a hydrogen atom.)

[0018] Examples of alkyl groups having 1 to 4 carbon atoms include methyl, ethyl, propyl, and butyl groups. The alkyl group may be linear or branched.

[0019] Examples of alkenyl groups having 2 to 3 carbon atoms include vinyl groups, allyl groups, and propenyl groups (1-propenyl groups).

[0020] The ring formed by the bonding of R1 and R2 to each other can be a saturated ring structure containing a nitrogen atom of the amino group (a nitrogen atom that is a ring constituent atom) and a sulfoxide atom that forms a sulfoxide group as ring constituent atoms. For example, a saturated six-membered ring can be formed in which the nitrogen atom and the sulfur atom are located at positions 1 and 4, respectively. In this saturated six-membered ring, the nitrogen atom and the sulfur atom are bonded cyclically through two pathways involving two carbon atoms each.

[0021] Particularly preferred sulfur-containing amino acids are compounds in formula (I) in which R2 is a hydrogen atom and R1 is one or more selected from a methyl group, an ethyl group, an allyl group, a propyl group, a propenyl group, and a butyl group, or in which R1 and R2 are bonded to each other to form a six-membered ring containing a sulfur atom and a nitrogen atom. More preferably, compounds in which R2 is a hydrogen atom and R1 is one or more selected from a methyl group, an ethyl group, a propyl group, a propenyl group, and a butyl group, or in which R1 and R2 are bonded to each other to form a six-membered ring containing a sulfur atom and a nitrogen atom are particularly preferred.

[0022] In particular, the sulfur-containing amino acid used in this embodiment is preferably at least one sulfoxide amino acid selected from isoalliin, methiin, and cycloalliin, from the viewpoint of excellent AGEs reduction effect, more preferably contains two or more selected from isoalliin, methiin, and cycloalliin, and most preferably contains all of isoalliin, methiin, and cycloalliin.

[0023] The agent of this embodiment preferably contains 0.1% by mass or more of sulfur-containing amino acids in its solid content, which is preferable for its high AGEs-reducing effect; more preferably 1% by mass or more; even more preferably 2% by mass or more; and particularly preferably 3% by mass or more. Furthermore, the AGEs-reducing agent of this embodiment preferably contains sulfur-containing amino acids in an amount of 50% by mass or less, more preferably 25% by mass or less, and particularly preferably 10% by mass or less, from the standpoint of enhancing functionality by including other components in its solid content and reducing the amount ingested. In particular, it is preferable that the amount of amino acids containing sulfoxide groups is above the upper limit and / or below the lower limit mentioned above. The solid content mentioned above refers to the total mass of all components excluding the solvent. The solvent referred to here may be one commonly used in pharmaceuticals, quasi-drugs, or food and beverages, and examples include organic solvents such as water and ethanol.

[0024] For example, if the agent of this embodiment contains isoalliin, it is preferable in that it is excellent in its decomposition effect on AGEs such as Glc-AGEs and MG-AGEs, and especially in its excellent effect in suppressing the formation of AGEs such as pentosidine and MG-H1. From this viewpoint, it is preferable that the agent of this embodiment contains 0.6% by mass or more of isoalliin in its solid content in terms of its high AGEs reduction effect, more preferably 1.0% by mass or more, and particularly preferably 1.5% by mass or more. Furthermore, it is preferable that the AGEs reducing agent of this embodiment contains isoalliin in an amount of 25% by mass or less in its solid content in terms of enhancing functionality by including other components and reducing the amount ingested, more preferably 15% by mass or less, and particularly preferably 10% by mass or less.

[0025] Furthermore, for example, if the agent of this embodiment contains cycloalliin, it is preferable in that it has excellent decomposition effects on AGEs such as Glc-AGEs, particularly pentosidine, and excellent inhibitory effects on the formation of AGEs. From this viewpoint, the AGEs reducing agent of this embodiment is preferable in that it contains 1.0% by mass or more of cycloalliin in its solid content in terms of high AGEs reducing effect, more preferably 1.5% by mass or more, and particularly preferably 2.0% by mass or more. In addition, the AGEs reducing agent of this embodiment is preferable in that it contains 25% by mass or less of cycloalliin in its solid content in terms of enhancing functionality by including other components and reducing intake, more preferably 15% by mass or less, and particularly preferably 10% by mass or less.

[0026] For example, when the AGEs-reducing agent of the present embodiment contains methiin, it is preferable in terms of excellent effect of decomposing AGEs such as Glc-AGEs, particularly pentosidine, and excellent effect of inhibiting AGEs production. From this perspective, the AGEs-reducing agent of the present embodiment preferably contains 0.2% by mass or more of methiin in its solid content in terms of high AGEs-reducing action, more preferably 0.4% by mass or more, and particularly preferably 0.6% by mass or more. In addition, the AGEs-reducing agent of the present embodiment preferably contains methiin in an amount of 10% by mass or less in its solid content, which is preferable in terms of achieving higher functionality by containing other components and reducing the intake amount, more preferably 7% by mass or less, and particularly preferably 5% by mass or less.

[0027] In the AGEs-reducing agent of the present embodiment, the total amount of isoalliin, cycloalliin and methiin in the sulfur-containing amino acids is preferably 60% by mass or more, more preferably 70% by mass or more, still more preferably 80% by mass or more, and particularly preferably 90% by mass or more.

[0028] One or more sulfur-containing amino acids selected from isoalliin, methiin and cycloalliin are known to have effects of improving male menopausal symptoms and mental health. Since the agent of the present invention contains the sulfur-containing amino acids, it can have the above-mentioned respective actions derived from the sulfur-containing amino acids in addition to the AGEs-reducing action, and thus has high industrial applicability. In addition, in the present specification, the solid content refers to components other than water.

[0029] As a method for measuring the content of the sulfur-containing amino acids, a high performance liquid chromatography method can be used. Examples of the HPLC conditions include the following. <HPLC Conditions> Detector: Ultraviolet absorption spectrophotometer (measurement wavelength 210 nm) Column: Shodex Asahipak NH2P-50 4E (4.6 mm ID × 250 mm) Column temperature: 40°C Mobile phase: 20mM NaH2PO4 + 100mM H3PO4 aq. / CH3 CN =19 / 81 Flow rate: 1.2ml / min Injection volume: 10μL

[0030] The sulfur-containing amino acids used in this embodiment may be chemically synthesized or naturally derived, but it is preferable that they be derived from plants of the genus Allium because they allow for the stable availability of a relatively large amount of sulfur-containing amino acids, are highly safe, and have a long history of safe consumption. Therefore, it is preferable that the agent of this embodiment contains processed products of Allium plants as sulfur-containing amino acids because they are readily available, highly safe, and have a long history of safe consumption. As for Allium plants, there are no particular limitations as long as they contain sulfur-containing amino acids, but examples include onions, leeks, green onions, chives, chives, garlic, wild garlic, shallots, and leeks. Of these, onions, green onions, and shallots are preferred because they are inexpensive and contain a large amount of sulfur-containing amino acids, and in particular, it is preferable to use sulfur-containing amino acids derived from onions because they have excellent AGEs reduction effects, are readily available, and have a long history of safe consumption. In this embodiment, the edible parts of these Allium plants are preferably used as a source of sulfur-containing amino acids. For example, the bulbs of onions, garlic, and shallots; the leaves of Welsh onions, chives, and leeks; and the leaves and pseudostems of leeks. Furthermore, since the outer skin of the Allium plants does not contain sulfur-containing amino acids, it is preferable to remove it beforehand and not use it as a source of sulfur-containing amino acids.

[0031] In this embodiment, it is also preferable to use an extract of Allium plants as a processed product of Allium plants as the sulfur-containing amino acid. Using an extract of Allium plants as the sulfur-containing amino acid is preferable because it allows for the reduction of AGEs at a low cost while ensuring safety and peace of mind with naturally derived ingredients, and because it is expected that the effects will be improved by combining multiple sulfur-containing amino acids.

[0032] As a method for producing extracts from Allium plants, known methods can be used. For example, one method involves heating the Allium plant to inactivate the degrading enzymes, then extracting with water or an aqueous ethanol solution, and subjecting the resulting solution to cation exchange chromatography. Alternatively, after heating the Allium plant, it can be crushed or otherwise liquefied and treated with a γ-glutamyl-binding cleaving enzyme, and the resulting enzyme-treated product can be subjected to ion exchange chromatography. Sulfur-containing amino acids adsorbed onto the ion exchange chromatography column are eluted, for example, using an alkaline eluent. The eluent can be either a strongly alkaline solution or a weakly alkaline solution, but is preferably a solution with a pH of 8 to 14. The obtained eluate may be used as is as an extract, but it is preferable to concentrate it using a known method or to further desalt it. Furthermore, if necessary, it may be subjected to drying, freeze-drying, solidification, liquefaction, granulation, or powdering.

[0033] The extract of Allium plants may be an Allium extract obtained by treating heated Allium plants with a γ-glutamyl-binding cleaving enzyme and subjecting the resulting enzyme-treated product to ion-exchange chromatography (hereinafter also referred to as "Step A"), and in particular, an extract obtained from onions via Step A may be used. The extract obtained in Step A is preferred because it has a particularly high AGEs reduction effect. The extract will be described in detail below.

[0034] Step A includes (1) heating the onions, (2) treating the heated onions with a γ-glutamyl binding cleavage enzyme, and (3) subjecting the resulting enzyme-treated product to ion-exchange chromatography.

[0035] In step A, the product obtained by using heated onions in the process of (2) is particularly excellent in reducing AGEs. From the viewpoint of even greater AGEs reduction effect, the heating conditions in the process of (1) are preferably such that the pressure is 1 atmosphere (0.10 M) Pa or higher. Similarly, the pressure is preferably 5 atmospheres (0.51 M) Pa or lower, and more preferably 2 atmospheres (0.21 MPa) or lower. Similarly, the temperature is preferably 40°C or higher, and more preferably 80°C or higher. Also, the temperature is preferably 150°C or lower, and more preferably 120°C or lower. The heating time is preferably 5 minutes or more, more preferably 15 minutes or more, preferably 120 minutes or less, and more preferably 40 minutes or less.

[0036] From the standpoint of reducing AGEs, the above heating is preferably performed on onions that have not been subdivided. The "unsubdivided" onions subjected to heating in this process may have been processed by cutting, dividing, crushing, puncturing, or damaging them. For example, in this embodiment, "unsubdivided" may mean a concept that allows for the entire onion to be divided into 2, 4, 8, or 16 parts, although this may vary depending on the part and size of the onion used.

[0037] In the present invention, in step (2) described above, the onions heated in step (1) are treated with a γ-glutamyl-binding cleaving enzyme. This results in a high AGEs reduction performance obtained in the process of this embodiment. The γ-glutamyl-binding cleaving enzyme used in this process can be any enzyme that has the activity to cleave the γ-glutamyl group from peptides or amino acids, such as γ-glutaminase, γ-glutamyltransferase, γ-glutamyltranspeptidase, and γ-glutamylpeptidase. These enzymes may be extracted from animals, plants, microorganisms, etc., or may be commercially available products. A commercially available product is glutaminase SD-C100S from Amano Enzyme Co., Ltd.

[0038] To ensure the above enzymatic reaction proceeds sufficiently, it is preferable to finely chop the onions heated in the above process (1) before the enzymatic treatment. The means of fine chopping are not particularly limited and include processes such as shredding, crushing, pulverizing, and grinding. These processes can be carried out by known means such as blenders, mixers, cutters, and mills. Furthermore, since the finely chopped onions are usually too viscous to be subjected to enzymatic treatment as is, it is preferable to dilute them with an aqueous liquid to about 1 to 100 times their mass. Examples of aqueous liquids include water, acidic water, and alkaline water. It is preferable that these aqueous liquids are solutions adjusted to the optimal pH or near the pH of the γ-glutamyl binding cleavage enzyme to be used later.

[0039] The conditions for treatment with the γ-glutamyl binding cleavage enzyme can be appropriately set depending on the optimal conditions for the enzyme, the type of onion used, the part used, the state of subdivision, etc. Generally, the amount of enzyme added is preferably 0.001 to 1% by mass, more preferably 0.01 to 0.1% by mass, relative to the total amount of onion.

[0040] The reaction conditions for the enzyme are preferably at the optimal pH for the enzyme, at a temperature of 15-65°C for 1-24 hours, more preferably at 35-60°C for 2-6 hours. After the completion of the enzyme treatment, it is preferable to inactivate the γ-glutamyl binding cleavage enzyme by heating or pH adjustment. If necessary, the reaction product obtained from the enzyme treatment may be filtered, centrifuged, or pressed to separate the resulting solution. The resulting solution may be further concentrated. The optimal pH for the enzyme is preferably 4.5-10.5.

[0041] In the above process (3), the reaction product obtained in the enzymatic treatment of process (2) is subjected to ion exchange chromatography. The ion exchange resin for this ion exchange chromatography can be any cation exchange resin, but a strongly acidic cation exchange resin is preferred. A strongly acidic cation exchange resin refers to an acidic cation exchange resin that is more acidic than a carboxylic acid type ion exchange resin, and examples include sulfonic acid type ion exchange resins and perfluoroalkanesulfonic acid type ion exchange resins. Among these, a sulfonic acid type strongly acidic cation exchange resin is more preferred as the ion exchange resin.

[0042] Commercially available ion exchange resins can be used, and suitable examples include Diaion® UBK-550, Diaion® SK1B (manufactured by Mitsubishi Chemical Corporation), Amberlite® IR120B, Amberlite® 200C, Dowex® MSC-1 (The Dow Chemical Company), Duolite C26 (Rohm and Haas), and Lewatit® SP-112 (LANXESS Distribution GmbH).

[0043] Ion exchange chromatography can be carried out according to the usual procedure. The enzyme-treated product obtained in the above process (2) is diluted with distilled water or buffer solution as needed to prepare the sample solution. Preferably, the sample solution is adjusted to pH 1-5 before being passed through the ion exchange resin. The sample solution is then passed through the ion exchange resin. After that, the ion exchange resin is washed with a washing solution such as distilled water, and then the amino acids adsorbed on the column are eluted using an alkaline eluent. Either a strongly alkaline solution or a weakly alkaline solution can be used as the eluent, but it is preferably a solution with a pH of 8-14. The obtained eluate may be used as is as onion extract, but it is desirable to concentrate it by a known method or to further desalt it. Further processing such as drying, freeze-drying, solidification, liquefaction, granulation, or powdering may be performed as needed.

[0044] For example, in the solid content of an extract of Allium plants (especially onion extract, hereinafter the same), the total amount of isoalliin, methiin, and cycloalliin is preferably 2% by mass or more, and more preferably 2% by mass or more and 50% by mass or less. The isoalliin content is preferably 0.6% by mass or more in the solid content of the Allium plant extract, the methiin content is preferably 0.2% by mass or more in the solid content of the Allium plant extract, and the cycloalliin content is preferably 1.0% by mass or more in the solid content of the Allium plant extract. Each example described later satisfies the above content ranges.

[0045] Furthermore, if the agent of this embodiment contains an extract of the Allium genus, the amount of the Allium genus extract is arbitrary as long as it can be an active ingredient, and the onion extract may account for 0.5% or more by mass, or 1% or more by mass, or 3% or more by mass of the solid content in the agent.

[0046] As shown in the examples described later, the sulfur-containing amino acids in this embodiment exhibit excellent AGEs-reducing effects. AGEs (Advanced Glycation End Products), as mentioned above, are a general term for products generated by the glycation reaction of proteins, and are also called advanced glycation end products or late-stage glycation products. AGEs accumulate in the body and are reported to cause or worsen various symptoms related to age-related chronic diseases such as diabetes, arteriosclerosis, dementia and other neurological disorders, decreased skin elasticity, osteoporosis, cataracts, and metabolic syndrome. The sulfur-containing amino acids used in this embodiment can prevent or suppress the progression of these symptoms through their excellent AGEs-inhibiting effect. In particular, the sulfur-containing amino acids in this embodiment not only have a preventative aspect of suppressing AGEs generation, but also have an AGEs-degrading effect, making them excellent anti-glycation agents. The AGEs reduction effect in this embodiment includes both an AGEs formation inhibitory effect and an AGEs degradation promoting effect. The AGEs reducing agent of this embodiment can be used as an AGEs formation inhibitor (AGEs production inhibitor) and / or an AGEs degradation promoting agent.

[0047] Examples of AGEs include glyoxal-derived AGEs.S GO-AGE S Glycol-AGEs, which are AGEs derived from glycolaldehyde, and glucose-derived AGEs. S Examples include Glc-AGEs, 3DG-AGEs derived from 3-deoxyglucosone, and MG-AGEs derived from methylglyoxal. GO-AGE S Examples include glyoxal lysine dimer (GOLD) and carboxymethylarginine. Examples of glyco-AGEs include carboxymethyllysine (CML). Examples of Glc-AGEs include pentosidine and crossrin. Pentosidine can also be produced from other sugars (pentoses), etc. Examples of 3DG-AGEs include pyraline, pyropyridine, imidazolone, and 3-deoxyglucosone lysine dimer (DOLD). Examples of MG-AGEs include methylglyoxal lysine dimer (MOLD), carboxyethyl lysine, algupyrimidine, and methylglyoxal-hydroimidazolon.

[0048] In particular, the agent of this embodiment is preferred to reduce Glc-AGEs and / or MG-AGEs, and especially preferred to reduce pentosidine and / or methylglyoxal-hydroimidazolon.

[0049] Among Glc-AGEs, pentosidine is a cross-linked AGE, formed when lysine and arginine residues are cross-linked by sugar. Because pentosidine is a cross-linked protein, it tends to harden protein structures and accumulates in proteins that are present in the body for relatively long periods. For these reasons, pentosidine is known as an indicator of chronic glycation and aging. As mentioned above, pentosidine is known to bind to collagen and cause a decrease in skin elasticity, and is also known to be associated with inflammation, malnutrition, chronic kidney disease, and poor clinical outcomes (e.g., PLOS ONE | DOI:10.1371 / journal.pone.0163826 October 4, 2016, p1-16).

[0050] Methylglyoxal (MG) is a glycation reaction intermediate with a dicarbonyl structure, and MG-AGEs are known to be associated with the development of diabetic microvascular complications, vascular disorders such as endothelial dysfunction, diabetic retinopathy, and diabetic nephropathy. Examples of MG-AGEs include methylglyoxal-hydroimidazoline (MG-H1, etc.), which is produced from arginine residues and methylglyoxal. MG-H1, one of the methylglyoxal-hydroimidazolines, is known to be rapidly and abundantly produced in the body among AGEs. High baseline serum MG-H1 levels are known to be associated with severe endothelial dysfunction, type 2 diabetes, and a history and onset of cardiovascular diseases (such as stroke and myocardial infarction) (Cardiovascular Diabetology 2023, Vol.147, No.22). Furthermore, MG-H1 levels are known to increase significantly with the progression of diabetic nephropathy (Int. J. Mol. Sci. 2023, Vol.152, No.24).

[0051] The AGEs reducing agent of this embodiment is expected to suppress the onset or progression of the diseases and phenomena mentioned above by reducing Glc-AGEs and MG-AGEs in the body, for example, in the blood and tissues.

[0052] In this embodiment, the AGEs reduction effect is preferably achieved by applying sulfur-containing amino acids to the reaction system that generates AGEs or to the generated AGEs, thereby reducing AGEs. Specifically, in this embodiment, the AGEs reduction effect is preferably achieved by using sulfur-containing amino acids to suppress the generation of AGEs or to promote their degradation. The AGEs reducing agent of this embodiment can be used as either an AGEs degradation accelerator or an AGEs generation inhibitor. The AGEs reduction effect in this embodiment may occur either internally or externally. For example, in the former case, when sulfur-containing amino acids are taken orally, most of them move from the small intestine into the bloodstream and then into the body from the bloodstream. Therefore, they can act on AGEs present in the blood and tissues. It can also be reasonably expected that they can act on reaction systems that produce AGEs in the blood and tissues (intracellular, skin, blood vessels, bones, eyes (lens), brain, hair, joints, kidneys, liver, etc.). As shown in the examples described later, sulfur-containing amino acids have AGEs degradation promoting and AGEs production inhibiting effects, and are therefore expected to contribute to AGEs reduction through these effects. In this embodiment, if the AGEs reduction effect is an AGEs degradation promotion effect, the degradation promotion effect includes the effect of reducing AGEs after contacting sulfur-containing amino acids with AGEs in a reaction system where a certain amount of AGEs have been formed (for example, in the blood or tissues in the body).

[0053] The AGEs reducing agent of this embodiment may contain arginine. Arginine is known to have anti-glycation effects. In this embodiment, the AGEs production inhibitory effect obtained is further improved by using arginine in combination with a sulfur-containing amino acid. If the agent of this embodiment contains arginine, the amount is preferably 2% by mass or more, more preferably 2% by mass or more and 50% by mass or less, and even more preferably 2% by mass or more and 10% by mass or less, in the AGEs reducing agent.

[0054] Nutritional supplements containing arginine as the sole active ingredient tend to lead to high arginine intake. Arginine produces nitric oxide (NO) and widens blood vessels, so there are problems with arginine alone. If people with low blood pressure take too much, their blood vessels may widen excessively, and if people with high blood pressure take it together with blood pressure-lowering medication, their blood pressure may be lowered too much. In contrast, effective AGEs reducing agents, AGEs production inhibitors, and AGEs decomposition accelerators that contain arginine, as well as sulfur-containing amino acids in addition to arginine, are advantageous from a health perspective because they can maintain the ability to suppress AGEs production while suppressing arginine intake.

[0055] Methods of administering the agent in this embodiment include oral administration, transdermal administration, transmucosal administration, and enteral administration, with oral administration being particularly preferred. The agent of this embodiment can exert an AGEs-reducing effect when taken orally in an effective amount. The specific effective amount can be appropriately determined by those skilled in the art depending on the subject, age, weight, administration method, etc.

[0056] The dosage of the agent in this embodiment is generally preferably 5 to 1000 mg per day for adults, and more preferably 7 to 100 mg, as sulfur-containing amino acids. The agent in this embodiment can be taken continuously. Continuous intake means, for example, taking it for at least one day a week for three weeks or more, and preferably taking it for at least one day a week for one month or more.

[0057] When the agent of this embodiment is an extract of Allium plants, the amount to be taken is preferably 0.1 to 5000 mg per day for adults, and more preferably 10 to 500 mg, as the solid content of the Allium plant extract.

[0058] The agent of this embodiment can be used as a pharmaceutical, quasi-drug, or food / beverage for animals, including humans, or for the manufacture of such products. The agent of this embodiment may be directly administered or ingested by animals, including humans, as a pharmaceutical, quasi-drug, or food / beverage, or it may be added to or blended with food / beverage or animal feed such as pet food and used as food / beverage or animal feed for reducing AGEs, etc. In the latter case, the method of adding or blending the agent of this embodiment into food / beverage or animal feed is not particularly limited. For example, the agent of this embodiment may be directly blended into raw materials before the manufacture of food / beverage or animal feed, added during the manufacturing process of food / beverage or animal feed, or added to the manufactured food / beverage or animal feed. The aforementioned "food and beverages" refers to anything that can be consumed by humans as food, and includes not only general food and beverages, including so-called health foods, but also health functional foods such as Foods for Specified Health Uses and Foods with Nutrient Function Claims as defined by the Ministry of Health, Labour and Welfare's Health Functional Foods System, as well as supplements, etc. The aforementioned "animal feed" refers to substances given as food to animals other than humans (animals raised by humans), such as livestock, poultry, and fish. Examples include livestock feed and pet food.

[0059] When the agent of this embodiment is used as a pharmaceutical or quasi-drug, the agent of this embodiment may contain sulfur-containing amino acids as the active ingredient alone, or it may further contain pharmaceutically acceptable carriers, or it may further contain other active ingredients or pharmacological components to the extent that the AGEs formation inhibitory effect and AGEs decomposition promoting effect of sulfur-containing amino acids are not impaired. Examples of such carriers include excipients, coating agents, binders, bulking agents, disintegrants, surfactants, lubricants, diluents, dispersants, buffers, osmotic pressure adjusters, pH adjusters, emulsifiers, preservatives, stabilizers, antioxidants, colorants, ultraviolet absorbers, humectants, thickeners, activity enhancers, bactericides, flavoring agents, odoring agents, etc.

[0060] When the agent of this embodiment is used as a pharmaceutical or quasi-drug, it can be administered in any dosage form. The dosage form may be oral or parenteral. For example, oral dosage forms include solid forms such as tablets, coated tablets, granules, powders, and capsules, as well as liquid forms such as elixirs, syrups, and suspensions. Parenteral dosage forms include injection, infusion, transdermal, transmucosal, nasal, enteral, inhalation, suppositories, boluses, and patches. Of these, oral dosage forms are preferred.

[0061] When the agent of this embodiment is used as a food or beverage, it may contain sulfur-containing amino acids as the active ingredient alone, or it may further contain various additives used in the manufacture of food and beverages, to the extent that the AGEs formation inhibitory effect and AGEs decomposition promoting effect of sulfur-containing amino acids are not impaired. Examples of such additives include various oils and fats, herbal medicines, amino acids, polyhydric alcohols, natural polymers, vitamins, dietary fiber, surfactants, purified water, excipients, stabilizers, pH adjusters, antioxidants, sweeteners, flavor components, acidulants such as organic acids, stabilizers, flavors, colorants, fragrances, etc.

[0062] Examples of food and beverages include oral medications (gum, candy, etc.), processed seafood products such as kamaboko and chikuwa, livestock products such as sausages and ham, bread, Western-style sweets, Japanese-style sweets, noodles such as fresh noodles, Chinese noodles, boiled noodles, and soba, seasonings such as sauces, soy sauce, dips, sugar, honey, powdered syrup, and starch syrup, spices such as curry powder, mustard powder, and pepper powder, jams, marmalades, chocolate spreads, pickles, side dishes, furikake, or processed vegetables and fruits such as canned and bottled vegetables and fruits, dairy products such as cheese, butter, and yogurt, beverages such as miso soup, soups, fruit juices, vegetable juices, whey drinks, soft drinks, and alcoholic beverages, and other general food and beverages such as health foods.

[0063] This embodiment includes a package comprising a packaging body and the aforementioned agent of this embodiment contained in the packaging body, or a pharmaceutical product, quasi-drug, food or beverage, or animal feed containing the same. The packaging can contain the agent, pharmaceutical, quasi-drug, food or beverage, or animal feed of this embodiment, and can be printed with ingredient labels, etc., but its form and material are not particularly limited. Examples of the form of the packaging include a box or a bag. Examples of the material of the packaging include paper, plastic, woven fabric, metal, etc. The packaging clearly displays various information, such as the content of sulfur-containing amino acids in the agent, food, beverage, or animal feed of this embodiment contained within the packaging. The method of presenting information in such packaging is not particularly limited. For example, 1) it may be printed on the outer or inner surface of the packaging; 2) information such as AGEs reduction uses may be printed on a printing medium, such as printing paper, enclosed inside the packaging along with the food, beverage, or animal feed; or 3) a QR code (registered trademark) containing an internet URL or information that allows access to a site introducing the agent, food, beverage, or animal feed of this embodiment may be included in the packaging or enclosed printing medium, and the information may be presented by accessing that URL.

[0064] Furthermore, this embodiment can provide a method for reducing AGEs by applying sulfur-containing amino acids to a reaction system that generates AGEs or to the generated AGEs. This method does not require medical treatment of humans and may, for example, be a cosmetic method.

[0065] Next, a second embodiment of the present invention will be described. In the second embodiment, instead of sulfur-containing amino acids, an onion extract obtained by treating heated onions with a γ-glutamyl bond cleaving enzyme and subjecting the resulting enzyme-treated product to ion-exchange chromatography is used as an essential component. In this case, the AGEs reducing agent of this second embodiment (hereinafter also collectively referred to as "the agent of this second embodiment") will be used. All descriptions of "the agent of this embodiment" given in the first embodiment can be applied to the description of the agent of the second embodiment. For example, the method for producing the onion extract in the second embodiment corresponds entirely to the description of step A described above. Furthermore, the description of the onion extract in the second embodiment corresponds entirely to the description of the onion extract in the first embodiment. Moreover, the effects of inhibiting AGEs production, promoting AGEs degradation, and reducing AGEs, as described in the first embodiment, are all exhibited by the onion extract in the second embodiment in the same way as in the first embodiment, and the descriptions of inhibiting AGEs production, promoting AGEs degradation, and reducing AGEs, as described in the first embodiment, all correspond to the second embodiment. Therefore, the onion extract in the second embodiment can be used not only as an AGEs reducing agent, but also as an AGEs production inhibitor and an AGEs degradation promoter. Additionally, the agent in the second embodiment can contain sulfur-containing amino acids. All descriptions of preferred types and content of sulfur-containing amino acids in the first embodiment can be applied to the description of the agent in the second embodiment.

[0066] Although the present invention has been described above based on its preferred embodiments, the present invention is not limited to what is described above. [Examples]

[0067] The present invention will be described in more detail below with reference to examples. However, the scope of the present invention is not limited to these examples.

[0068] (Manufacturing Example 1) 5000g of harvested onions were washed and depeeled, then whole and uncut were heat-treated in a water bath at 95°C for 20 minutes. The heat-treated onions were crushed using a mixer (Oster Co., Ltd.) and dispersed by adding water to a total volume of 50% by mass. To the resulting dispersion, a glutaminase preparation (Glutaminase SD-C100S; Amano Enzyme Co., Ltd.), one of the γ-glutamyl bond cleaving enzymes, was added as an enzyme at a concentration of 0.025% by mass relative to the total volume of onions in the solution. The mixture was reacted at 60°C for 120 minutes, and after the reaction was complete, the mixture was heated to inactivate the enzyme. The resulting reaction solution was centrifuged, filtered by suction, and then freeze-dried to obtain a crude onion extract. Distilled water was added to the crude onion extract described above to obtain a sample solution with a solid content of 5% by mass. A column packed with 500 mL of strong cation exchange resin (Diaion SK1B, manufactured by Mitsubishi Chemical Corporation, a sulfonic acid type strong acid cation exchange resin) regenerated with hydrochloric acid was prepared. After adjusting the pH of the sample solution to 1-5 using an acid such as hydrochloric acid, the sample solution was passed through the column of the strong cation exchange resin, and then the sample solution remaining in the column was washed out with 3000 mL of distilled water. Subsequently, 1000 mL of a 5% by mass aqueous solution of sodium hydroxide (pH 14) was passed through to eluate the components adsorbed on the ion exchange resin. Further elution of the liquid remaining in the column was performed with 2000 mL of distilled water. The eluate eluted with the 5% by mass aqueous solution of sodium hydroxide and the eluate eluted with distilled water were combined, neutralized to pH 7 with hydrochloric acid, concentrated using an evaporator, desalted, and dried by freeze-drying to obtain a powdered onion extract. The onion extract contained 3.3% by mass of isoalliin, 1.1% by mass of methiin, and 3.2% by mass of cycloalliin.

[0069] <Test 1> (Example 1) The AGEs reduction performance of isoalliin, methiin, cycloalliin, and the onion extract obtained in Production Example 1 was investigated using the following method.

[0070] 1. Inhibition of pentosidine production (Reagents used) Disodium hydrogen phosphate (anhydrous), special grade reagent (manufactured by Junsei Chemical Co., Ltd.) Sodium dihydrogen phosphate (anhydrous), special grade reagent (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) Human serum albumin (HAS, manufactured by Sigma-Aldrich) D(+)-glucose (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) Sodium borohydride (manufactured by Tokyo Chemical Industry Co., Ltd.) 100 w / v% trichloroacetic acid solution (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) Isoalliin (manufactured by Nagara Science Co., Ltd.) Methiine (manufactured by Funakoshi Co., Ltd.) Cycloalliin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.)

[0071] (Experimental process) 0.2M phosphate buffer (pH 7.4) was used. The sample solution was prepared by mixing and dissolving onion extract, isoalliin, methiin, cycloalliin, or aminoguanidine hydrochloride with purified water in the reaction solution (Solution (A)) after a 10-fold dilution, to the concentrations shown in Table 1. The amount of onion extract used was adjusted so that the amount of isoalliin in the onion extract reached the concentration shown in Table 1. Human serum albumin solution was prepared by arranging an aqueous solution of human serum-derived albumin at a concentration of 40 mg / mL. A glucose solution was prepared by 1 M aqueous solution of D(+)-glucose. For the glycation reaction, a solution (A) was prepared by mixing the sample solution, human serum albumin solution, glucose solution, and phosphate buffer in a ratio of 1:2:2:5, and the mixture was reacted at 60°C for 4 days. Solution (B) was prepared in the same manner as the preparation of solution (A) (a solution containing all of the sample solution, protein solution, glucose solution, and sample solution), except that purified water was added instead of the sample solution. Solution (B) was then reacted at 60°C for 4 days in the same manner as solution (A). After the saccharification reaction, sodium borohydride was added to the saccharification reaction solution to a concentration of 200 mM to protect the amino groups. 1 mL of 20% by mass trichloroacetic acid was added, and after standing, the supernatant was removed by centrifugation. Then, 1 mL of 5% by mass trichloroacetic acid was added to the precipitate, and after standing, the supernatant was removed by centrifugation. The precipitate was suspended in 100 μL of purified water, and then 100 μL of 6N hydrochloric acid was added. Hydrolysis was performed at 105°C for 18 hours. Pretreatment was performed using a Monospin AG column (GL Sciences), and the amount of pentosidine was measured by HPLC. The rate of pentosidine production inhibition (%) was calculated using the following formula. Aminoguanidine hydrochloride (AG) was used as a positive control for the inhibitory effect on glycation reactions. The results are shown in Table 1. The pentosidine concentration in the control group after the test was calculated to be 11 nM. Generation suppression rate (%)={(BA) / B}×100 A: Amount of pentosidine produced in (A) B: Amount of pentosidine produced in (B)

[0072] (HPLC conditions) Column:InertSustain AG (GL Sciences, 3 μm, 4.6 mm ID × 100 mm) Eluent: A) 0.1% HCOOH in H2O B) CH3CN A / B = 100 / 0 (v / v, same applies below) - (0-8 min) - 50 / 50 - (8-10 min) - 100 / 0 - (10-18 min) Step gradient Flow Rate: 1.0 mL / min Col. Temp.: 20℃ Detection: FL EX 325 nm, Em 385 nm Sample Size: 20 μL

[0073] (Pentosidine production inhibition rate) [Table 1]

[0074] As shown in Table 1, when aminoguanidine hydrochloride was included at a concentration of 5 mM in the positive control, the inhibition rate of pentosidine production was negative. However, when each sulfur-containing amino acid or onion extract containing them was used, the inhibition rate of pentosidine production improved. From the results in Table 1, it can be seen that the present invention provides a reduction effect by inhibiting the production of AGEs.

[0075] <Exam 2> (Example 2) 2. Pentosidine degradation (Reagents used) Disodium hydrogen phosphate (anhydrous), special grade reagent (manufactured by Junsei Chemical Co., Ltd.) Sodium dihydrogen phosphate (anhydrous), special grade reagent (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) Human serum-derived albumin (manufactured by Sigma-Aldrich) D(+)-glucose (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) Sodium borohydride (manufactured by Tokyo Chemical Industry Co., Ltd.) 100 w / v% trichloroacetic acid solution (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) Isoalliin (manufactured by Nagara Science Co., Ltd.) Methiine (manufactured by Funakoshi Co., Ltd.) Cycloalliin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.)

[0076] (Experimental process) 0.2M phosphate buffer (pH 7.4) was used. The sample solution was prepared by mixing and dissolving onion extract, isoalliin, methiin, cycloalliin, or aminoguanidine hydrochloride with purified water in the reaction solution (Solution (A)) after a 10-fold dilution, to the concentrations shown in Table 2. The amount of onion extract used was adjusted so that the amount of isoalliin in the onion extract reached the concentration shown in Table 2. Human serum albumin solution was prepared by arranging an aqueous solution of human serum-derived albumin at a concentration of 40 mg / mL. As a glucose solution, a 1M aqueous solution of D(+)-glucose was prepared. For the glycation reaction, human serum albumin solution, glucose solution, and phosphate buffer were mixed in a ratio of 2:2:5 and reacted at 60°C for 4 days. Subsequently, solution (A) was prepared by adding the sample solution to a volume of 1 / 10 of the total volume (total volume after adding the sample solution), and reacted at 37°C for 24 hours. Solution (B) was prepared in the same manner as the preparation of solution (A) (solution obtained by adding the sample solution to the saccharification reaction sample), except that purified water was added instead of the sample solution. Solution (B) was then reacted at 37°C for 24 hours, in the same manner as solution (A). Subsequently, sodium borohydride was added to the saccharification reaction solution to a concentration of 200 mM to protect the amino groups, and 1 mL of 20% by mass trichloroacetic acid was added. After standing, the mixture was centrifuged and the supernatant was removed. Then, 1 mL of 5% by mass trichloroacetic acid was added to the precipitate, and after standing, the mixture was centrifuged and the supernatant was removed. After removing the supernatant, the precipitate was suspended in 100 μL of purified water, and then 100 μL of 6N hydrochloric acid was added. Hydrolysis was performed at 105°C for 18 hours, and pretreatment was performed using a Monospin AG column (GL Sciences). The amount of pentosidine was measured by HPLC under the same conditions as in Example 1. The pentosidine degradation rate (%) was calculated using the following formula. The results are shown in Table 2. The pentosidine concentration in the control group after the test was calculated to be 11 nM. Decomposition rate (%)={(BA) / B}×100 A: Amount of pentosidine produced in (A) B: Amount of pentosidine produced in (B)

[0077] (Pentosidine degradation rate) [Table 2]

[0078] As described above, in this experiment, the amount of pentosidine was reduced in solution (A), which was held at 37°C for 24 hours after adding the test substance (sample solution), compared to solution (B), which was prepared by reacting sugar and protein at 60°C for 4 days and then adding purified water instead of the test substance (sample solution) and holding it at 37°C for 24 hours. It appears that the formation of pentosidine proceeds sufficiently during the reaction of sugar and protein at 60°C for 4 days. Therefore, it is understood that in this reaction, the degradation of the generated AGEs is promoted by the action of the test substance, sulfur-containing amino acid, or onion extract containing it.

[0079] <Exam 3> (Example 3) 3. Methylglyoxal-hydroimidazolon (MG-H1) decomposition (Reagents and equipment used) Disodium hydrogen phosphate (anhydrous), special grade reagent (manufactured by Junsei Chemical Co., Ltd.) Sodium dihydrogen phosphate (anhydrous), special grade reagent (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) Methylglyoxal (MG) solution (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) Arginine (manufactured by Tokyo Chemical Industry Co., Ltd.) Bovine serum albumin (BSA, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) Isoalliin (manufactured by Nagara Science Co., Ltd.) (Experimental process) 0.2M phosphate buffer (pH 7.4) was used. The sample solution was prepared by mixing isoalliin with water in solution (A) to achieve the concentration shown in Table 3 when diluted tenfold, and then dissolving it. A bovine serum albumin (BSA) solution was prepared using purified water as the solvent, with a concentration of 50 mg / mL. The arginine solution was prepared using purified water as the solvent to a concentration of 40 mg / mL. The methylglyoxal solution was adjusted to a concentration of 20 mM using purified water as the solvent. For the saccharification reaction, BSA solution or arginine solution, methylglyoxal solution, phosphate buffer, and purified water were mixed in a ratio of 1:1:22:12 and reacted at 37°C for 16 hours. Subsequently, when reacted with BSA solution, the saccharification reaction solution was centrifuged using an Amicon Ultra 30kDa filter (Millipore), and the sample on the filter was collected. On the other hand, when reacted with arginine solution, the saccharification reaction solution was used as is. The sample solution was then added to the solution to a volume of 1 / 10 of the total volume (total volume after adding the sample solution), and the reaction was carried out at 37°C for 24 hours. Solution (B) was prepared in the same manner as the preparation of solution (A) (solution obtained by adding the sample solution to the saccharification reaction sample), except that purified water was added instead of the sample solution. Solution (B) was then reacted at 37°C for 24 hours, in the same manner as solution (A). For the above solutions (A) and (B), the absorption wavelength at 450 nm was measured using a microplate reader with the OxiSelect® methylglyoxal (MG) competitive ELISA kit (Cell Biolabs), and the MG-H1 concentration was determined, and the amount of MG-H1 generated was calculated. Specifically, the following procedure was followed. (1) 50 μL of MG-BSA solution diluted 10-fold with PBS was added to a 96-well plate with MG-Conjugate immobilized on it, and the plate was infused at room temperature for 10 minutes. (2) 50 μL of Anti-MG Antibody was added and allowed to permeate at room temperature for 1 hour. (3) Washing was performed using 250 μm of 1×WASH Buffer. This process was repeated three times. (4) 100 μL of Secondary Antibody-HRP Conjugate was added and allowed to permeate at room temperature for 1 hour. (5) Washing was performed using 250 μL of 1×WASH Buffer. This process was repeated 5 times. (6) Add 100 μL of Substrate Solution and allow the color reaction to develop for about 10 minutes. (7) 100 μL of Stop Solution was added to stop the color development reaction. (8) The absorption at a wavelength of 450 nm was measured using a macro plate reader, and the amount of AGEs produced by the MG-BSA / MG-arginine solution was examined in comparison to the color development of the attached MG-BSA Standard, and the MG-H1 degradation rate was calculated. The results are shown in Table 3. The MG-H1 decomposition rate was calculated using the following formula. Decomposition rate (%)={(BA) / B}×100 A: Amount of AGEs produced in (A) B: Amount of AGEs produced in (B)

[0080] (MG-H1 decomposition rate) [Table 3]

[0081] As described above, in this experiment, the amount of MG-H1 was reduced in solution (A), which was held at 37°C for 24 hours after adding the test substance, compared to solution (B), which was prepared by reacting methylglyoxal with arginine or protein at 37°C for 16 hours, and then adding purified water instead of the test substance (sample solution) and holding it at 37°C for 24 hours. Reacting methylglyoxal with arginine or protein at 37°C for 16 hours converts methylglyoxal to MG-H1. Therefore, it is understood that the reaction during the subsequent 24-hour holding at 37°C accelerates the decomposition of the generated AGEs due to the action of the sulfur-containing amino acid, which is the test substance, or the onion extract containing it. In particular, in the case of the BSA solution, since the sugar and protein are separated by centrifugation of the saccharification reaction solution using an Amicon Ultra 30kDa filter (Millipore), it is thought that further saccharification does not proceed, making the acceleration of decomposition even clearer.

[0082] <Exam 4> (Example 4) The onion extract obtained above was investigated for its ability to suppress AGEs production using the following method.

[0083] (Reagents used) Sodium hydrogen phosphate dodecahydrate, special grade reagent (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) Sodium dihydrogen phosphate dihydrate, special grade reagent (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) Methylglyoxal solution (manufactured by Sigma-Aldrich) Bovine serum albumin (Fraction V), endotoxin-free (Merck Millipore)

[0084] (Experimental process) Using the onion extract prepared in the same manner as described above (reagents used) and in Example 1, a reaction solution with the following composition was prepared and incubated at 37°C for 16 hours. (MG-BSA solution) (composition) 50 mM phosphate buffer (pH 7.4) 2.5 mg / mL Bovine serum albumin 1 mM methylglyoxal 4.5 mg / mL Onion extract

[0085] (Quantitative process) The OxiSelect® Methylglyoxal (MG) Competitive ELISA Kit (manufactured by CELL BIO LABS) was used. Specifically, the procedure was as follows: (1) 50 μL of MG-BSA solution diluted 10-fold with PBS was added to a 96-well plate with MG-Conjugate immobilized on it, and the plate was infused at room temperature for 10 minutes. (2) 50 μL of Anti-MG Antibody was added and allowed to permeate at room temperature for 1 hour. (3) Washing was performed using 250 μm of 1×WASH Buffer. This process was repeated three times. (4) 100 μL of Secondary Antibody-HRP Conjugate was added and allowed to permeate at room temperature for 1 hour. (5) Washing was performed using 250 μL of 1×WASH Buffer. This process was repeated 5 times. (6) Add 100 μL of Substrate Solution and allow the color reaction to develop for about 10 minutes. (7) 100 μL of Stop Solution was added to stop the color development reaction. (8) The absorption at a wavelength of 450 nm was measured using a macro plate reader, and the amount of AGEs produced in the MG-BSA solution was determined by comparing it to the color development of the attached MG-BSA Standard. The production rate and suppression rate of MG-H1 were then calculated. The results are shown in Table 4. The production rate and inhibition rate of MG-H1 were calculated using the following formula. Production rate (%) = A / B × 100 (%) Generation suppression rate (%)={(BA) / B}×100 A: Amount of AGEs produced in (Comparative Examples 1, 2, or Example 4) B: Amount of AGEs produced in (Comparative Example 1) The amount of AGEs produced in Example 4 was 8.0 (μg / mL). (The production rate of MG-H1 was the average of two consecutive samples.)

[0086] (Comparative Example 1) Except for the absence of onion extract, the amount of AGEs produced was investigated and the MG-H1 production inhibition rate was calculated in the same manner as in Example 4. The amount of AGEs produced was 28.8 μg / mL.

[0087] (Comparative Example 2) Instead of using onion extract, we used the same amount of arginine as that contained in onion extract (2.55% by mass) to similarly investigate AGEs production and calculate the MG-H1 production inhibition rate.

[0088] Table 4 shows the MG-H1 production inhibition rate for Example 4. A higher value indicates a higher ability to inhibit AGEs production.

[0089] (MG-H1 production suppression rate) [Table 4]

[0090] As shown in Table 4, the onion extract used in this invention exhibits excellent inhibitory effects on AGEs production. Furthermore, as can be seen from the comparison between Comparative Example 2 and Example 4, the onion extract had a higher MG-H1 production inhibitory effect compared to using the same amount of arginine as that contained in the onion extract.

Claims

1. An AGEs reducing agent containing sulfur-containing amino acids.

2. An AGEs reducing agent containing sulfur-containing amino acids as its active ingredient.

3. The AGEs reducing agent according to claim 1 or 2, wherein the sulfur-containing amino acid has a sulfoxide group.

4. An AGEs reducing agent according to claim 1 or 2, which suppresses the formation of AGEs or promotes their degradation.

5. The AGEs reducing agent according to claim 1 or 2, wherein the sulfur-containing amino acid is a compound represented by the following formula (I). 【Chemistry 1】 (In the formula, R １ and R ２ They bond to each other to form a ring, or R １ R is an alkyl group having 1 to 4 carbon atoms or an alkenyl group having 2 to 3 carbon atoms. ２ (This is a hydrogen atom.)

6. R ２ is a hydrogen atom and R １ is one or more selected from methyl group, ethyl group, allyl group, propyl group, propenyl group, and butyl group, or R １ and R ２ The AGEs reducing agent according to claim 5, wherein the two atoms bond to each other to form a six-membered ring containing a sulfur atom and a nitrogen atom.

7. The AGEs reducing agent according to claim 1 or 2, wherein the sulfur-containing amino acid is one or more selected from isoalliin and cycloalliin.

8. An AGEs reducing agent according to claim 1 or 2, comprising a plant of the genus Allium as a source of sulfur-containing amino acids.

9. The AGEs reducing agent according to claim 8, wherein the plant of the Allium genus includes onion extract.

10. The AGEs reducing agent according to claim 9, wherein the onion extract is obtained by a step of treating heated onions with a γ-glutamyl-binding cleaving enzyme and subjecting the resulting enzyme-treated product to ion-exchange chromatography.

11. The AGEs reducing agent according to claim 1 or 2, wherein the AGEs are one or more selected from pentosidine or methylglyoxal-hydroimidazolon.

12. A method for reducing AGEs by applying sulfur-containing amino acids to a reaction system that generates AGEs or to the generated AGEs.

13. A method for reducing AGEs according to claim 12, wherein the sulfur-containing amino acid is used to suppress the formation of AGEs or to promote their degradation.

14. The heated onions were treated with a gamma-glutamyl binding cleavage enzyme, An AGEs reducing agent comprising an onion extract obtained in a step of subjecting the resulting enzyme-treated product to ion-exchange chromatography.

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