Advanced glycation end products inhibitor

Abstract

An advanced glycation endproduct production inhibitor according to the present invention contains a natto bacteria culture extract as an active ingredient. Since containing a natto bacteria culture extract derived from a food material as the main ingredient, the present invention can be ingested safely without any concern of side effects. In addition, the present invention can be mass-produced and is easily distributed on the market.

Description

[Technical Field]

[0001] The present invention relates to an agent for inhibiting the production of advanced glycation end products (AGEs). [Background technology]

[0002] The incidence of type 2 diabetes has been steadily increasing in recent years, and its complications (diseases that induce vascular disorders such as nephropathy, retinopathy, and peripheral neuropathy) are a major factor in shortening healthy life expectancy. If left untreated, these complications can become fatal, for example, nephropathy leading to dialysis, retinopathy leading to blindness, and neuropathy leading to vascular disorders such as painless myocardial infarction and gangrene.

[0003] Furthermore, it has been reported that "AGEs" (Advanced Glycation Endproducts) are involved in kidney damage in type 2 diabetes (e.g., Non-Patent Documents 1 to 5), and much research is being conducted into their relationship to the onset and progression of the disease. AGEs are also called "advanced glycation end products" or "advanced glycation end products," and refer to modified proteins formed by oxidative / nonoxidative reactions initiated by a non-enzymatic reaction between protein and sugar (glycation reaction). The series of reactions involved in the production of AGEs is also known as the Maillard reaction.

[0004] The increase in AGEs in type 2 diabetes may be related to increased oxidative stress caused by metabolic disorders such as hypertension, hyperlipidemia, obesity, and hyperinsulinemia, as well as hyperglycemia.

[0005] For this reason, there is a need to establish a technology that can suppress the production of AGEs in the body and inhibit the onset and progression of the above-mentioned type 2 diabetes and other conditions. [Prior art documents] [Patent documents]

[0006] [Non-Patent Document 1] Kang J et al., Acta Diabetol. 2005 Jun;42(2):110-6 [Non-patent document 2] Miyata T et al., Kidney Int. 1998 Feb;53(2):416-22 [Non-patent document 3] Friedman EA, Nephrol Dial Transplant. 1999;14 Suppl 3:1-9 [Non-patent document 4] Christensen EI et al., Nat Rev Mol Cell Biol. 2002, Apr;3(4):256-66 [Non-Patent Document 5] Gugliucci A et al., Diabetologia. 1996 Feb;39(2):149-60 Summary of the Invention [Problem to be solved by the invention]

[0007] The present invention is intended to solve the above problems, and its object is to provide an advanced glycation end product formation inhibitor that can effectively inhibit the formation of AGEs in the body and is highly safe. [Means for solving the problem]

[0008] The present invention is an agent for inhibiting the formation of advanced glycation end products, which contains a Bacillus subtilis natto culture extract as an active ingredient.

[0009] In one embodiment, the Bacillus subtilis natto culture extract contains nattokinase.

[0010] In one embodiment, the Bacillus subtilis natto culture extract is in the form of a dry powder.

[0011] In one embodiment, the vitamin K2 content in the Bacillus natto culture extract is 1 μg or less per 1 g of the dry mass of the Bacillus natto culture extract.

[0012] In one embodiment, the Bacillus subtilis natto culture extract has a nattokinase activity of 100 FU / g to 80,000 FU / g.

[0013] In one embodiment, the advanced glycation endproducts formation inhibitor of the present invention is an oral formulation.

[0014] The present invention also provides a method for inhibiting the production of advanced glycation end products in a living organism, the method comprising administering a Bacillus subtilis natto culture extract as an active ingredient to the living organism.

[0015] In one embodiment, the administration to the living body is performed orally.

[0016] The present invention also relates to use of a composition containing a Bacillus subtilis natto culture extract as an active ingredient for inhibiting the production of advanced glycation end products. [Effects of the Invention]

[0017] According to the present invention, by containing a Bacillus subtilis natto culture extract derived from a food material as a main ingredient, it can be safely ingested without the risk of side effects. In addition, it can be mass-produced and easily distributed to the market. [Brief explanation of the drawings]

[0018] [Figure 1] FIG. 1 shows the experimental design carried out in Example 3. [Figure 2] 1 is a photograph showing the surface of a thin section of a tissue specimen in the pathological tissue evaluation of a renal nephron performed in Example 3. [Figure 3] 1 is a graph showing the glycation state of renal tubules scored based on vacuolation of epithelial cells and positive PAS staining in the pathological tissue evaluation of renal nephrons performed in Example 3. [Figure 4] 1 is a graph showing the results of measuring plasma advanced glycation end products (AGEs) concentrations performed in Example 3. DETAILED DESCRIPTION OF THE INVENTION

[0019] The advanced glycation endproducts formation inhibitor of the present invention contains a Bacillus subtilis natto culture extract as an active ingredient.

[0020] The term "advanced glycation end products (hereinafter sometimes referred to as AGEs)" as used herein refers collectively to modified proteins formed by oxidative / nonoxidative reactions initiated by a nonenzymatic reaction (glycation reaction) between proteins and sugars. Furthermore, the term "inhibition of the formation of advanced glycation end products" as used herein encompasses both inhibiting the increase in the amount of AGEs produced by, for example, the Maillard reaction in vivo or in vitro and keeping it roughly constant, as well as inhibiting the production of the AGEs themselves and reducing the amount produced.

[0021] The natto bacteria culture extract is an extract obtained from a culture medium of natto bacteria (typically Bacillus subtilis natto), a type of Bacillus subtilis. The natto bacteria may be any microorganism capable of producing nattokinase, which will be described later. The natto bacteria may be natto bacteria isolated from commercially available natto.

[0022] The natto Bacillus subtilis culture extract may be in the form of, for example, a dry powder, a liquid, or a paste, but is preferably in the form of a dry powder because it is easy to handle and commercially available.

[0023] Furthermore, a natto Bacillus culture extract in which the strong natto odor, which is characteristic of natto, has been reduced or removed is preferably used, because this makes it easier for more people to ingest. For example, in order to reduce or remove the natto odor, it is preferable that the natto Bacillus culture extract is one in which certain impurities have been removed from the extract in advance. Specifically, the natto Bacillus culture extract is one in which impurities having a molecular weight of preferably 1000 or less, more preferably 20,000 or less, have been removed.

[0024] Bacillus subtilis natto culture extract contains nattokinase as the main component.

[0025] Nattokinase (NK) is a substance with fibrinolytic activity found in the viscous substance of natto. It is a subtilisin-family serine protease consisting of 275 amino acids. Nattokinase has been shown to exhibit high thrombolytic activity in vivo in experimental systems, improve peripheral blood flow, inhibit platelet aggregation, lower blood pressure in hypertension, and enhance the immune system. Furthermore, in in vitro experimental systems, it has been shown to have higher fibrinolytic activity than plasmin, degrade and inactivate PAI-1, and increase tissue plasminogen activator (tPA).

[0026] The Bacillus subtilis natto culture extract has a certain nattokinase activity, which is expressed in units of FU (fibrinolytic units) per gram and can be detected by determining whether or not lytic plaques form on a fibrin plate, according to the method described in Experientia, Vol. 43, p. 1110 (1987), for example.

[0027] The Bacillus natto culture extract of the present invention preferably has a nattokinase activity of 100 FU / g to 80,000 FU / g, more preferably 4000 FU / g to 80,000 FU / g. If the nattokinase activity of the Bacillus natto culture extract is less than 100 FU / g, a contradiction may arise in that a larger amount of the Bacillus natto culture extract is required to achieve the desired effect of efficiently inhibiting the production of advanced glycation end products.

[0028] Note that the nattokinase activity in commercially available natto is usually only 20 FU / g to 40 FU / g. In contrast, the nattokinase activity per given amount of Bacillus natto culture extract is much higher than that of commercially available natto. Therefore, the advanced glycation end products formation inhibitor of the present invention containing the Bacillus natto culture extract can have a high nattokinase activity that cannot be obtained by using commercially available natto.

[0029] Usually, when Bacillus subtilis natto is cultured, the culture solution contains both nattokinase, an active factor in the thrombolytic system, and vitamin K, an active factor in the thrombus coagulation system. Vitamin K is known to counteract the effects of nattokinase, and is a component that should preferably be removed in the present invention. For this reason, in the present invention, the content of vitamin K2 contained in the Bacillus subtilis natto culture extract is adjusted to preferably 1 μg or less, more preferably 0.1 μg or less, per 1 g of dry mass of the Bacillus subtilis natto culture extract.

[0030] Vitamin K2 contained in the natto bacteria culture extract can be removed by, for example, combining a natto bacteria culture solution containing vitamin K2 with an aqueous chitosan solution to adsorb vitamin K2 to the chitosan, and then filtering off the chitosan with the adsorbed vitamin K2, as described in JP 2006-325597 A.

[0031] The advanced glycation end products formation inhibitor of the present invention may contain an oily substance to improve the dispersibility of the Bacillus subtilis natto culture extract. The oily substance is preferably one that does not solidify at temperatures of, for example, 50°C or lower, preferably 40°C or lower. Examples of oily substances include soybean oil, glycerin fatty acid esters, beeswax, rapeseed oil, jojoba oil, palm oil, coconut oil, and combinations thereof.

[0032] The content of the oily substance in the advanced glycation end products formation inhibitor of the present invention is not particularly limited. For example, an appropriate amount can be selected by those skilled in the art based on the nattokinase activity of the Bacillus subtilis natto culture extract used in combination.

[0033] The advanced glycation end products formation inhibitor of the present invention may also contain other additives in addition to the Bacillus subtilis natto culture extract. Examples of such other additives include additives that are commonly used in the food and / or pharmaceutical fields, such as excipients, stabilizers, lubricants, binders, flow agents, disintegrants, sweeteners, flavors, and colorants, as well as combinations thereof.

[0034] Examples of excipients include erythritol, sorbitol, xylitol, maltitol, lactose, sucrose, trehalose hydrate, reduced maltose syrup, crystalline cellulose, dextrin, corn starch, potato starch, wheat starch, rice starch, partially pregelatinized starch, sodium bicarbonate, calcium phosphate anhydrous, calcium hydrogen phosphate hydrate, calcium phosphate tribasic, calcium carbonate, precipitated calcium carbonate, calcium silicate, and calcium lactate, as well as combinations thereof.

[0035] Examples of stabilizers include dextrin, α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, sucrose fatty acid esters, hydroxypropyl-β-cyclodextrin, hydroxypropyl cyclodextrin, and mannitol, and combinations thereof.

[0036] Lubricants include, for example, stearic acid, magnesium stearate, calcium stearate, sodium stearyl fumarate, sucrose fatty acid esters, hydrogenated oils, glycerin, glycerin fatty acid esters, carnauba wax, and talc, and combinations thereof.

[0037] Examples of binders include pullulan, pectin, sodium alginate, gum arabic, guar gum, agar, starch syrup, hydroxypropyl cellulose, pregelatinized starch, polyvinylpyrrolidone, carboxyvinyl polymer, polyvinyl alcohol, ammonioalkyl methacrylate copolymer, ethyl cellulose, carboxyvinyl polymer, carboxymethyl ethyl cellulose, hypromellose, polyvinyl alcohol-polyethylene glycol graft copolymer, polyvinyl alcohol-acrylic acid-methyl methacrylate copolymer, methyl cellulose, beeswax, macrogol, and methacrylic acid copolymer, and combinations thereof.

[0038] Flow agents include, for example, silicon dioxide, light anhydrous silicic acid, magnesium aluminometasilicate, and fumed silica, and combinations thereof.

[0039] Disintegrants include, for example, hydroxypropyl cellulose, carboxymethyl cellulose, carmellose calcium, corn starch, partially pregelatinized starch, croscarmellose sodium, carboxymethyl starch sodium, and crospovidone, and combinations thereof.

[0040] Examples of sweeteners include aspartame, saccharin, stevia, amacha powder, sucralose, acesulfame potassium, sucrose, sorbitol, reduced maltose syrup, saccharin, licorice, and thaumatin, and combinations thereof.

[0041] Flavoring agents include, for example, orange, vanilla, strawberry, yogurt, menthol, fennel oil, cinnamon oil, spruce oil, peppermint oil, and green tea powder, and combinations thereof.

[0042] Coloring agents include, for example, iron sesquioxide, yellow iron sesquioxide, black iron oxide, titanium oxide, talc, Food Yellow No. 4, Food Yellow No. 4 Aluminum Lake, Food Yellow No. 5, Food Red No. 2, Food Red No. 3, Food Red No. 102, Food Blue No. 1, methylene blue, carmine, and riboflavin, and combinations thereof.

[0043] The content of the additive in the advanced glycation end products formation inhibitor of the present invention is not particularly limited. For example, an appropriate amount can be selected by those skilled in the art based on the nattokinase activity of the Bacillus subtilis natto culture extract used in combination.

[0044] The dosage of the advanced glycation endproducts formation inhibitor is preferably 1000 to 4000 FU (fibrinolytic units), more preferably 2000 to 4000 FU per day for an adult (body weight 60 kg to 65 kg), and may be administered in divided doses per day.

[0045] The inhibitor of advanced glycation endproducts formation of the present invention is, for example, an oral preparation that can be ingested by animals such as humans, pets, livestock, poultry, and farmed fish, and is used, for example, as a food or drink, feed (feeding agent), or pharmaceutical product itself or as one of their constituent materials.

[0046] The inhibitor of the formation of advanced glycation end products of the present invention may have any dosage form, and examples of such dosage forms include powder, granules, pills, tablets, and liquids. When such an inhibitor of the formation of advanced glycation end products is used as a food or drink, examples of the food or drink include general foods; supplements; health functional foods such as foods for specified health uses, foods with nutrient functions, and foods with functional claims; soft drinks; tea drinks; coffee drinks; processed milk; dairy drinks; soy milks; and alcoholic beverages.

[0047] The advanced glycation end product formation inhibitor of the present invention contains, as an active ingredient, a Bacillus subtilis natto culture extract that can be obtained from natto, which is a common food product as described above. Therefore, safety for ingestion by living organisms (e.g., humans) is fully guaranteed. Furthermore, it can be easily ingested by people of a wide range of ages, from infants to adults and the elderly.

[0048] The inhibitor of the advanced glycation end products formation of the present invention can suppress the onset and progression of, for example, type 2 diabetes by, for example, suppressing the formation of AGEs in the body. Alternatively, the inhibitor of the advanced glycation end products formation of the present invention can also be used as a blood AGE concentration-lowering agent for lowering the blood AGE concentration when ingested by humans, for example. [Example]

[0049] Next, the present invention will be further explained by way of examples, but the present invention is not limited to these examples.

[0050] Example 1: Preparation of low-dose nattokinase-containing feed (NK-L) We received a nattokinase (NK)-containing natto bacteria culture extract (NSK-SD) manufactured by Nippon Biological Science Institute Co., Ltd. This product was obtained by filtering the culture extract obtained from fermentation of natto bacteria to remove impurities with a molecular weight of 20,000 or less, and then spray-drying it to produce a powder (containing dextrin as a stabilizer). The powder contained 3.4% by mass of nattokinase. The nattokinase activity was 20,000 FU / g.

[0051] Next, we commissioned CLEA Japan Inc. to prepare a diet containing 0.6% of the above-mentioned Bacillus subtilis natto culture extract (NSK-SD) per 1 g of normal solid feed (CE-2 manufactured by CLEA Japan Inc.). This resulted in a low-dose nattokinase-containing diet (NK-L) containing 0.2 mg of nattokinase (NK) per 1 g of feed. )of Got it.

[0052] Example 2: Preparation of high-dose nattokinase-containing feed (NK-H) A high-dose nattokinase-containing feed (NK-H) containing 0.6 mg of nattokinase (NK) per 1 g of feed was prepared in the same manner as in Example 1, except that the preparation of the feed containing 1.8% of the above-mentioned Bacillus subtilis natto culture extract (NSK-SD) per 1 g of normal solid feed (CE-2 manufactured by CLEA Japan, Inc.) was outsourced to CLEA Japan, Inc. )of Got it.

[0053] Example 3: Creation of an STZ-induced type II diabetes model animal and administration of each nattokinase-containing feed The following animals and reagents were used to evaluate the low-dose nattokinase-containing feed (NK-L) and the high-dose nattokinase-containing feed (NK-H) obtained in Examples 1 and 2.

[0054] (1)Animals All experimental procedures were carried out in accordance with laws and regulations related to experiments, such as the Act on Welfare and Management of Animals, and the Hiroshima International University Experimental Animal Guidelines, and animal experiments were conducted in an ethical manner.

[0055] Six-week-old male Spragur-Dawly (SD) rats were purchased from Japan SLC Co., Ltd. and housed at a density of three rats per cage in a breeding room (Hiroshima International University Kure Campus Animal Breeding Facility) under conditions of 23±1°C, 55±5% humidity, and a 12-hour light-dark cycle (light 8:00–20:00, dark 20:00–8:00). After 10 days of acclimation, animals were used in this experiment when they reached a body weight of 240–280 g. During acclimation, animals were fed normal solid diet (CE-2, CLEA Japan Inc.) and provided with water ad libitum using an automatic water valve.

[0056] (2) Reagents Streptozotocin (STZ), diethyl ether (special grade), sodium citrate buffer solution (pH 4.5), cholesterol E-Test Wako, laboratory assay TM Triglycerides were purchased from Fujifilm Wako Pure Chemical Industries, Ltd. Novo-Heparin Injection 10,000 units / 10 mL was purchased from Mochida Pharmaceutical Co., Ltd. and used. A self-testing glucose kit, Diasensor, purchased from Arkray, Inc. was used. Oxi Select™ Advanced Glycation End Product (AGE) Competitive ELISA Kit was purchased from Cosmo Bio Co., Ltd. and used. 1 mL of Sureeads Protein G Magnetic Beads was purchased from Bio-Lad and used. Glucose was purchased from Merck.

[0057] (3) Experimental method The experiment was carried out based on the plan shown in Figure 1. Specifically, the experiment was carried out as follows.

[0058] Male SD rats (8 weeks old) were acclimated for 10 days and divided into three groups (n=6) using a stratified random sampling method based on two parameters, fasting blood glucose and body weight, to ensure uniform mean and variance values ​​for each parameter. Streptozocin (STZ), adjusted to pH 4.5 with sodium citrate, was administered intraperitoneally once at a dose of 55 mg / 3 mL / kg body weight, and the rats were then raised for 14 days.

[0059] Immediately after STZ administration, the control group was given normal solid feed (CE-2, manufactured by CLEA Japan, Inc.), the low-dose NK group was given the low-dose nattokinase-containing feed (NK-L) (0.2 mg NK / g CE-2) obtained in Example 1, and the high-dose NK group was given the high-dose nattokinase-containing feed (NK-H) (0.6 mg NK / g CE-2) obtained in Example 2, and they were allowed to freely consume these for 14 days.

[0060] Only animals that showed a fasting blood glucose level of 200 mg / dL or higher on the seventh day after STZ administration and became diabetic were allowed to continue the experiment. This was based on the empirical rule that in animals whose blood glucose levels did not reach abnormal levels on the seventh day after STZ administration, no increase in blood glucose levels could be observed until the end of the experiment. The animals were housed in stainless steel rabbit cages at a density of three per cage. Food intake was measured for each cage three times a week during the experimental period.

[0061] Blood glucose levels were measured and blood samples were collected immediately before STZ administration, and on days 7 and 14 after the start of test substance administration. Immediately before STZ administration, and 7 and 14 days after the start of test substance administration (the final day of the study), rats were fasted for 15 hours (7:00 PM to 10:00 AM) and 500 μL of heparinized blood was collected from the tail vein. The blood that flowed out before hemostasis (30 μL) was used as a sample for blood glucose measurement using a self-test glucose meter (GT-1670, Arkray, Inc.). The heparinized blood was centrifuged (1500 × g, 10 minutes) in a high-speed refrigerated microcentrifuge (MX-100, Tommy Seiko Co., Ltd.). The obtained plasma was stored at -80°C until use as a sample for measuring blood biochemical parameters.

[0062] Immediately after blood collection, the rats were euthanized by exsanguination via abdominal aorta resection, and the left kidney was removed and fixed in 10% formalin buffer (pH 6.8). Male SD rats (8 weeks old) were kept under normal conditions for 14 days, and the left kidney was removed in the same manner as above and used as a normal specimen.

[0063] (4) Measurement of blood biochemical markers The plasma obtained in the above experiment (4) was used as a sample to measure AGEs.

[0064] AGEs were measured using the Oxi Select™ Advanced Glycation End Product (AGE) Competitive ELISA Kit.

[0065] (5) Histopathological evaluation of the kidney On day 14 after STZ administration, the left kidneys of each group were excised, dehydrated using an automatic processor, and embedded in paraffin. Histological specimens (4 μm) were stained with hematoxylin-eosin (HE), periodic acid-methenamine silver (PAM), and Peritic Acid Schiff (PAS) to observe the renal corpuscles and tubules.

[0066] (6) Results (Body weight and food intake) Table 1 shows the weight gain and food intake of the control group, the low-dose NK-fed group fed the low-dose nattokinase-containing feed (NK-L) obtained in Example 1 (hereinafter also referred to as the "NK-L-fed group of Example 1"), and the high-dose NK-fed group fed the high-dose nattokinase-containing feed (NK-H) obtained in Example 2 (hereinafter also referred to as the "NK-H-fed group of Example 2").

[0067] [Table 1]

[0068] As shown in Table 1, the average weight gain over the 14 days from the start of test substance administration was approximately 2 g / day in all groups, with no difference between the groups. There was also no difference in food intake.

[0069] (Blood biochemistry data) The blood glucose levels of the normal group, the control group, the NK-L-fed group of Example 1, and the NK-H-fed group of Example 2 are shown in Table 2.

[0070] [Table 2]

[0071] As shown in Table 2, blood glucose levels in all groups were above 200 mg / dL on day 7 after STZ administration (day 7 after the start of test substance administration), and on day 14 rose to 326.75±17.0 mg / dL in the control group, 419.8±23.3 mg / dL in the NK-L-fed group of Example 1, and 401±31.0 mg / dL in the NK-H-fed group of Example 2, which were significantly higher than the normal value (67.11±1.3 mg / dL). However, there was no significant difference in the values ​​between the groups.

[0072] (Pathological evaluation of renal nephron tissue) The left kidneys were excised from the rats of each group and the surrounding nephrons were subjected to histopathological evaluation.

[0073] Specifically, the left kidney of each rat was removed from each group, dehydrated using an automatic processor, and embedded in paraffin as described in (4) Experimental Methods above. For the normal group (N), SD male rats (8 weeks old) were fed a normal chow diet (CE-2, CLEA Japan, Inc.) for 14 days and processed as described above. The tissue specimens were thinly sectioned (4 μm) and stained with hematoxylin-eosin (HE), periodic acid-methenamine silver (PAM), and Peritic Acid Schiff (PAS) to observe the renal corpuscle and renal tubules (Figure 2).

[0074] Furthermore, the glycation status of the renal tubules was scored based on the vacuolation of epithelial cells and positive PAS staining. Evaluation was based on a 5-point scale (-: no change, +: slight change, ++: moderate change, +++: severe change, ++++: marked change). Testing was performed using the Mann-Whitney U test, and P values ​​were calculated (*: P<0.05). The results are shown in Figure 3.

[0075] As shown in Figure 2, in the renal corpuscles (HE staining; left lane), no lesions were observed around the renal corpuscles in any of the control group, the NK-L-fed group of Example 1, and the NK-H-fed group of Example 2, and no abnormalities were observed compared to the normal group. On the other hand, in the renal tubules (HE staining; center lane), vacuolation of epithelial cells was observed in all groups compared to the normal group, as indicated by the arrows. In the renal tubules (PAS staining; right lane), glycation of epithelial cells (positive PAS staining: glycogen deposition) was observed in all groups compared to the normal group, as indicated by the arrows. Furthermore, as shown in Figure 3, glycation rated as ++ was observed in all individuals in the control group and the NK-L-fed group of Example 1, while mild cases rated as + were observed in the NK-H-fed group of Example 2. Mild cases were observed in three of six cases in the NK-H-fed group of Example 2, and the mean score was significantly lower than that of the control group.

[0076] (Plasma advanced glycation end products (AGEs) concentration) The plasma advanced glycation end products (AGEs) concentration was measured for each of the normal value group, the control group, the NK-L feeding group of Example 1, and the NK-H feeding group of Example 2.

[0077] Specifically, measurements were performed using stored plasma samples from rats in each group as described in (4) Experimental Methods above, and the measured values ​​were expressed as mean ± standard error. Statistical analysis was performed using Dunnett's method, and the difference with the control group was compared (*: P<0.05). The results are shown in Figure 4.

[0078] The plasma AGE concentration obtained was 4.12±0.48 μg / dL in the control group, which was significantly increased compared to the normal value group (2.06±0.19 μg / dL). On the other hand, the concentration was 2.81±0.75 μg / dL in the NK-L fed group of Example 1 and 1.30±0.36 μg / dL in the NK-H fed group of Example 2. Compared to the control group, the value decreased depending on the NK dose, and was significantly lower in the NK-H fed group of Example 2. [Industrial Applicability]

[0079] The present invention is useful in technical fields such as the food field and the pharmaceutical field.

Claims

1. It consists of natto bacteria culture extract, The Bacillus subtilis natto culture extract has a nattokinase activity of 20,000 FU / g to 80,000 FU / g, and is an advanced glycation end products formation inhibitor.

2. 2. The advanced glycation end products formation inhibitor according to claim 1, wherein the Bacillus subtilis natto culture extract contains nattokinase.

3. The advanced glycation endproducts formation inhibitor according to claim 1 , wherein the Bacillus subtilis natto culture extract is in the form of a dry powder.

4. 2. The advanced glycation endproducts formation inhibitor according to claim 1, wherein the vitamin K2 content contained in the Bacillus subtilis natto culture extract is 1 μg or less per 1 g of the dry mass of the Bacillus subtilis natto culture extract.

5. The advanced glycation endproducts formation inhibitor according to claim 1 , which is an oral formulation.

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