Nutritional composition for preventing bronchitis

Abstract

To find out from breast milk components a component for preventing bronchitis and to provide a new nutritional composition.SOLUTION: The disclosure relates to a nutritional composition for preventing bronchitis, the nutritional composition including N-acetylneuraminic acid, 6'-sialyllactose, α2,6-disialyl N-linked glycans, and a disialyl N-linked glycan with a specific structure as active ingredients. Since the nutritional composition is effective not only for infants but also for elderly individuals at high risk of bronchitis and patients with underlying diseases, it is also effective as a nutritional composition for preventing bronchitis when incorporated into foods and nutritional compositions for adults, including the elderly.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a nutritional composition for preventing bronchitis. The present invention relates to a nutritional composition for preventing bronchitis, which contains lanuraminic acid as an active ingredient. [Background technology]

[0002] Bronchitis is an inflammation caused by a virus or bacteria infecting the airways (bronchi) where the trachea branches. Bronchitis is caused by respiratory syncytial virus (RSV), rhinovirus, and adenovirus. It is a viral infection such as a virus, coronavirus, or influenza virus, but Bacterial pathogens such as coplasma and Bordetella pertussis can also be causes. After infection, bacteria such as pneumococcus and influenza bacteria can cause secondary infections, leading to serious illness. It can also become

[0003] Infants and young children are at high risk of contracting bronchitis because their immune systems are not yet fully developed. Infants up to about 3 months old have narrow bronchi, so they have a weak ability to expel phlegm, which can lead to the spread of inflammation. There is a high risk of developing serious illness. Furthermore, elderly people whose immune systems are declining are also at high risk of bronchitis. If the infection worsens, it can progress to pneumonia, a serious and life-threatening condition.

[0004] Therefore, preventing bronchitis is a vital health issue for everyone, from infants to the elderly. It is important for living a healthy social life, and various studies are being conducted on its treatment and prevention methods. Regarding treatment after contracting bronchitis, anti-influenza virus Treatment includes the administration of medicines, various antibiotics, cough suppressants, and expectorants. Patent Document 1 describes an antiulcer agent or It discloses a bifidobacteria growth promoter, and also has antiviral properties against influenza viruses and rotaviruses. It has also been shown to be effective in treating infections caused by viruses such as flu and herpes. However, the patent document does not show any efficacy in preventing bronchitis. Although it has been shown to prevent viral diarrhea, rotavirus is a causative virus of bronchitis. Therefore, it has not been shown to prevent bronchitis. There are no effective legal measures, and the only thing that can be done is to encourage people to wash their hands and gargle. .

[0005] By the way, breast milk contains various components that reduce the risk of infection. It is known (Patent Document 2). In particular, carbohydrates, mainly oligosaccharides, contained in breast milk It has been reported that this compound inhibits the stage at which the virus attaches to host cells (Patent Document 3). In searching for an active ingredient that prevents the onset of bronchitis, the present inventors have investigated the However, whether it is possible to specifically prevent the onset of bronchitis itself is unclear. It is unclear what components in breast milk may be able to prevent bronchitis. It was not possible to do so. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Patent No. 3930559 [Patent Document 2] Patent No. 2514375 [Patent Document 3] Patent No. 3789146 Summary of the Invention [Problem to be solved by the invention]

[0007] The present inventors have discovered a component in breast milk that prevents the onset of bronchitis. If possible, the ingredient can be incorporated into infant nutritional compositions to prevent infants from contracting bronchitis. We thought that we could provide a nutritional composition for infants. It is also an effective ingredient for the elderly and those with underlying diseases who are at high risk of bronchitis. It is formulated into foods and nutritional compositions for adults and the elderly to provide nutritional support to prevent the onset of bronchitis. It was thought that it might be possible to supply it as a composition. The object of the present invention is to find a component from breast milk that can prevent the onset of bronchitis, and The objective of the present invention is to provide a nutritional composition. [Means for solving the problem]

[0008] The present inventors have investigated the carbohydrates in breast milk in order to search for components in breast milk that prevent the onset of bronchitis. Statistical analysis of the relationship between the amount of ingredients and the history of bronchitis in infants to prevent bronchitis As a result of extensive research into new ingredients, we discovered that breast milk contains N-acetylneuraminic acid, 6'-acetylcysteine, and Allyl lactose, α2,6 disialo N-linked glycans, and disialo N-linked glycans with specific structures The inventors have found that the compound is effective in preventing bronchitis, and have completed the present invention. That is, the present invention relates to a method for producing a sialic acid derivative comprising N-acetylneuraminic acid, 6'-sialyllactose, α2, 6. Contains disialo N-linked glycans and disialo N-linked glycans with specific structures as active ingredients The present invention relates to a nutritional composition for preventing the onset of bronchitis. Specifically, the present invention has the following configuration.

[0009] <1> A nutritional formula containing N-acetylneuraminic acid as an active ingredient for preventing bronchitis Finished product. <2> The N-acetylneuraminic acid is a protein, a peptide, an oligosaccharide, or a lipid. Binds to one or more <1> The nutritional composition described in <3> The binding mode of N-acetylneuraminic acid is α2,6 bond <1> or <2> written in The nutritional composition described above. <4> N-acetylneuraminic acid-containing sugar chains exist as 6'-sialyllactose <3> The nutritional composition described in <5> N-acetylneuraminic acid-containing glycans are N-linked glycans <3> Nutritional composition described in Finished product. <6> The nutritional composition according to <5>, wherein the sugar chain containing N-acetylneuraminic acid is an α2,6 disialo N-linked sugar chain. <7> The α2,6 disialo N-linked glycan has the following structure (1) or (2): <6> The nutritional composition described in (1)Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα 1-6)Manβ1-4GlcNAcβ1-4GlcNAc (2)Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα 1-6)Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc <8> It is for infants and young children, <1> from <7> The nutritional composition according to any one of the preceding claims. <9> <1> from <8> The nutritional composition for preventing bronchitis according to any one of , Food and drink for preventing bronchitis. <10> <1> from <8> The nutritional composition for preventing bronchitis according to any one of A medicine for preventing bronchitis. [Effects of the Invention]

[0010] According to the present invention, a nutritional composition containing N-acetylneuraminic acid as an active ingredient can improve the tracheal This ingredient can prevent the onset of bronchitis. It is also effective for elderly people with high blood pressure and those with underlying diseases, so it is suitable for adults and elderly people with high blood pressure. It is also possible to prevent the onset of bronchitis. [Brief explanation of the drawings]

[0011] [Figure 1] This shows the effect of N-acetylneuraminic acid concentration in breast milk on the incidence of bronchitis in infants. [Figure 2] This shows the effect of 6'-SL concentration in breast milk on the incidence of bronchitis in infants. [Figure 3] This shows the effect of the concentration of disialo N-linked glycan A in breast milk on the incidence of infant bronchitis. [Figure 4] This shows the effect of the concentration of disialo N-linked glycan B in breast milk on the incidence of bronchitis in infants. [Figure 5] This shows the effect of the concentration of α2,6 disialo N-linked glycans in breast milk on the incidence of infant bronchitis. DETAILED DESCRIPTION OF THE INVENTION

[0012] (N-acetylneuraminic acid) The N-acetylneuraminic acid of the present invention, even in a free state, is an oligosaccharide that can be easily synthesized from other N-acetylneuraminic acid may be bound to a sugar. Alternatively, it may be bound to a protein or lipid. Acids can be derived from milk, eggs, bird's nests, genetically modified bacteria fermentation, or synthetic. It may be any of synthetic, semi-synthetic or natural products. Also, natural materials containing natural products may be used as a supply source as it is and incorporated into the nutritional composition of the present invention. Extracted by known methods from natural materials containing N-acetylneuraminic acid For example, milk-derived The original glycomacropeptide and sialylglycopeptide, which is a condensed glycan portion of glycomacropeptide, were Chido, etc. In the present invention, the binding mode of N-acetylneuraminic acid is preferably α2,6 binding. α2,6-linked N-acetylneuraminic acid is an oligosaccharide or glycoprotein sugar. linked to galactose or N-acetylgalactosamine residues in the α2,6 bond N-acetylneuraminic acid, Neu5Acα2-6Gal structure, or Neu5Ac It refers to a compound that has an α2-6GalNAc structure.

[0013] (6'-sialyllactose) In the present invention, the sugar chain containing N-acetylneuraminic acid is 6'-sialyllactose ( It is desirable that 6'-sialyllactose ( Hereafter, it may be simply referred to as 6'-SL) is derived from milk and genetically modified bacteria fermentation method. It may be a synthetic product, a semi-synthetic product, or a natural product. Natural materials such as these may be used as a source of nutrients and incorporated into the nutritional composition of the present invention. The natural product is a product obtained by a known method from a natural material containing 6'-sialyllactose. It means an extracted product, a crudely purified product, or a product that has been further purified from these.

[0014] (N-linked glycans) In the present invention, the sugar chain containing N-acetylneuraminic acid exists as an N-linked sugar chain. It is desirable that the N-linked sugar chain containing N-acetylneuraminic acid of the present invention is It may be in the form of a peptide or a protein. It can be derived from eggs, meat, microorganisms, genetically modified bacteria, or synthetic products. It may be a natural product, a semi-synthetic product, or a natural product. It may be incorporated into the nutritional composition of the present invention as a supply source as it is. Those extracted by known methods from natural materials containing bound glycans, and those roughly purified The term "purified" refers to a product obtained by further refining the product.

[0015] (α2,6 disialo N-linked glycan) The α2,6 disialo N-linked glycan in the present invention is an N-linked glycan that reduces the risk of bronchitis. The α2,6 disialo N-linked sugar chain of the present invention is, even in a free state, It may be bound to a peptide and / or protein. They may be derived from meat, microorganisms, or genetically modified bacteria fermentation processes, and may be synthetic, semi-synthetic, or Natural products may be used as a source of natural materials. The natural product may be an α2,6 disialo N Those extracted by known methods from natural materials containing bound glycans, and those roughly purified The term "purified" refers to a product obtained by further refining the product.

[0016] The α2,6 disialo N-linked sugar chain in the present invention is specifically Neu5Acα2-6Galβ1- 4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6)Manβ1-4GlcNAcβ1-4G lcNAc (hereinafter simply referred to as disialo N-linked glycan A) and Neu5Acα2-6Galβ1-4Gl cNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6)Manβ1-4GlcNAcβ1-4(Fuc In addition to α1-6)GlcNAc (hereinafter simply referred to as disialo N-linked glycan B), for example, Neu5Acα2-6Galβ1-4(Fucα1-3)GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2M anα1-6)Manβ1-4GlcNAcβ1-4GlcNAc and Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5 Acα2-6Galβ1-4(Fucα1-3)GlcNAcβ1-2Manα1-6)Manβ1-4GlcNAcβ1-4GlcNAc, Neu5Acα 2-6Galβ1-4GlcNAcβ1-2(Neu5Acα2-6Galβ1-4GlcNAcβ1-4)Manα1-3(Galβ1-4GlcNAcβ1 -2Manα1-6)Manβ1-4GlcNAcβ1-4GlcNAc, etc., which have two molecules of Neu5Acα2,6 at the non-reducing end. It refers to N-linked glycans with a glycan bond.

[0017] The nutritional composition of the present invention contains an effective amount of N-acetylneuraminic acid to prevent bronchitis. The preferred range of the amount required per 100g of solids of the nutritional composition is as follows: show. It is preferable that the N-acetylneuraminic acid content is 170 mg / 100 g solid or more. More preferably, the amount is 189 mg / 100 g solid or more. In this case, the prepared milk preferably contains 221 mg / L or more of N-acetylneuraminic acid. It is preferably 246 mg / L or more, and more preferably 246 mg / L or more. In addition, the nutritional composition of the present invention contains 46.9 mg of 6'-sialyllactose per 100 g of solids. It is preferable that the solid content is 104 mg / 100 g or more, and more preferably 104 mg / 100 g or more. It is preferable that the prepared milk contains 61.0 mg / L or more of 6'-SL. Preferably, it is 135 mg / L or more. Furthermore, the nutritional composition of the present invention contains α2,6 disialo N-linked glycans at a concentration of 0.198 μmol / It is preferable that the solid content is 100 g or more, and more preferably 0.338 μmol / 100 g solids or more. In addition, the amount of α2,6 disialoic acid in the prepared milk is 0.257 μmol / L or more. It is preferable that N-linked sugar chains are contained, and more preferably 0.440 μmol / L or more is. Furthermore, the nutritional composition of the present invention contained 0.0728 μmol / 10 disialo N-linked glycan A. It is preferable that the solid content is 0.0 ... In addition, the amount of disialo N-linked sugars in the prepared milk is 0.0947 μmol / L or more. It is preferable that chain A is contained, and more preferably, it is 0.313 μmol / L or more. In addition, the nutritional composition of the present invention contains disialo N-linked sugar chain B at 0.0935 μmol / 10 It is preferable that the solid content is 0.0 ... In addition, the amount of disialo N-linked glycans in the prepared milk is 0.122 μmol / L or more. It is preferable that B is contained, and more preferably, it is 0.245 μmol / L or more.

[0018] (nutritional compositions, food and beverages, pharmaceuticals) The present invention relates to a method for preventing bronchitis, which comprises administering to a patient an effective amount of N-acetylneuraminic acid or the like. The nutritional composition may be any composition containing an effective amount of N-acetylneuraminic acid or the like, and may contain the active ingredient In addition, it can contain proteins, fats, carbohydrates, vitamins and minerals, making it suitable for infants and young children. It is used as a nutritional composition for children as well as for adults and the elderly. Nutritional compositions for infants include infant formula and liquid formula, infant formula (full formula), Follow-up milk and growing-up milk), low birth weight infant formula, cow's milk These include allergen-free foods for those with allergies and lactose-free foods for those with lactose intolerance. . Nutritional compositions for adults and the elderly include adult formula, liquid formula, and milk. Allergen-free foods for allergies, lactose-free foods for lactose intolerance, supplements, etc. Examples include: In addition, the present invention provides a food and drink containing N-acetylneuraminic acid or the like for preventing bronchitis. As for the food and drink, it can be added to any food and drink that contains an effective amount of N-acetylneuraminic acid, etc. It may be mixed with other ingredients or added to the ingredients during the manufacturing process of food and beverages. Dairy products such as fermented milk, lactic acid bacteria drinks, butter, margarine, milk drinks, fruit juice drinks, soft drinks Beverages such as egg, jellies, candies, puddings, egg products such as mayonnaise, butter cakes Examples of suitable confectioneries and breads include, but are not limited to, breads and other sweets. The nutritional composition for preventing bronchitis of the present invention can be used as a raw material for pharmaceuticals. These can be produced in the form of tablets, capsules, powders, syrups, etc., by conventional methods. Therefore, the present invention provides a method for preventing bronchitis, which comprises administering N-acetylneuraminic acid or the like as an active ingredient. When formulating a pharmaceutical product containing the composition for use as an active ingredient, the excipients permitted for formulation shall be used. In addition to mixing ingredients such as steroids, stabilizers, and flavoring agents appropriately, The composition containing the compound can be dried as it is and used as a powder or dust. To the extent that the preventive effect is not impaired, excipients, binders, disintegrants, lubricants, flavorings, suspending agents, It is also possible to mix the drug with other drugs and other optional drugs to make a formulation. The formulations may be tablets, capsules, granules, powders, dusts, syrups, etc. [Example]

[0019] [Test Example 1] Various carbohydrate components were measured in 153 breast milk samples, and the risk of bronchitis in infants during the first year of life was assessed. The relationship between the incidence and non-presence of the disease was analyzed using logistic regression. The method and results are shown below.

[0020] 1. Method (1) Investigation of breast milk samples and infant infection history Snow Brand Beanstalk Co., Ltd. conducted a survey of Japanese breast milk maturity between 2014 and 2019. A prospective cohort study to investigate the relationship between pregnancy and maternal and child health status (UMIN ID 000015494 ) examined breast milk samples collected from 1,210 mothers and the health status of mothers and their children. The questionnaire results were used in this study. In addition, 200 samples were randomly selected from the 1,210 samples. The breast milk samples to be analyzed were collected between one and two months after birth. The questionnaire surveyed the health status of the infants, and the results were collected at two months and one year after birth. The parents filled out the questionnaire six times every other day, and the results were collected. The results were compiled based on whether or not a doctor diagnosed the mother with bronchitis within one year of giving birth. If the answer to the question "No" was unclear, the person was excluded from the survey. In the survey, 153 samples were properly answered regarding whether or not they were diagnosed with bronchitis. Of the 153 samples, 153 infants were diagnosed with bronchitis within one year of birth. The number of specimens that were examined was 18.

[0021] (2) Measurement of various carbohydrate components in breast milk (2-1) Measurement of total N-acetylneuraminic acid For each of the 153 breast milk samples analyzed, 100 μL was added to 800 μL of ultrapure water and 10 After adding 100 μL of 1N sulfuric acid, the mixture was heated at 80°C for 45 minutes. After adding 0.1N sodium hydroxide solution, the sample was filtered through an ultrafiltration membrane with a molecular weight cutoff of 10,000. The permeate was analyzed by HPLC. NEX ICS-5000DP system (Thermo Fisher Scientific Co., Ltd.) The column was a CarboPac PA1 column (Thermo Fisher Scientific) The mobile phase consisted of ultrapure water (liquid A) and 200 mM sodium hydroxide. Solution B, and 100 mM sodium hydroxide solution containing 600 mM sodium acetate The three solutions, solution B (solution C), and solution B, were passed through at a flow rate of 1 mL / min. Solution B was passed through at 10% and then solution B was linearly reduced to 37.5% within 10 minutes, and solution C was passed through at 25%. %. From 10 to 20 minutes, the concentration of solution B was 37.5% and that of solution C was 25%. From 20.1 to 25 minutes, 45% of solution B and 10% of solution C were passed through. From the calibration curve prepared using a standard N-acetylneuraminic acid, the total N-acetylneuraminic acid in each breast milk sample was The amount of cetylneuraminic acid was calculated.

[0022] (2-2) Measurement of oligosaccharides After adding 580 μL of ultrapure water to 20 μL of breast milk from 153 samples analyzed, The solution was treated with an ultrafiltration membrane with a molecular weight cutoff of 10,000, and the permeate was analyzed by HPLC. LC was performed using a DIONEX ICS-5000DP system (serving as a service) coupled with an electrochemical detector. The column was a CarboPac PA1 column. The mobile phase was ultrapure sorbent (Thermo Fisher Scientific). Water (Solution A), 200 mM sodium hydroxide solution (Solution B), and 600 mM sodium acetate The three solutions, 100 mM sodium hydroxide solution (solution C) containing sodium hydroxide, were passed through the tube at a flow rate of 1 mL / min. The solution B was passed through at 50% for the first 10 minutes, and then increased to 47.5% by 18 minutes. The concentration of solution B was increased linearly from 18 to 28 minutes. The solution B was passed through at 47.5% and 5% of solution C. From 28 to 32 minutes, the solution B increased linearly to 42%. Solution C was increased linearly to 16%. From 32 to 39 minutes, solution B was increased to 42%. After passing 16% of solution C, 30% of solution B and 40% of solution C were passed from 39 to 43 minutes. The concentrations of 2'-fucosyllactose, 3-fucosyllactose, and 3'-sialyllactose were lactose (hereinafter simply referred to as 3'-SL), 6'-sialyllactose (hereinafter , sometimes simply referred to as 6'-SL), lactodifucotetraose, lacto-N-tetraose ose, lacto-N-neotetraose, lacto-N-fucopentaose, lacto-N -Fucopentaose II, lacto-N-difucohexaose I, and lacto-N-difucohexaose The calibration curve prepared using the standard of cohexaose II indicated the content of each oligosaccharide in each breast milk sample. The total amount of these 11 types of oligosaccharides was calculated as "total oligosaccharides."

[0023] (2-3) Measurement of N-linked glycans For each of the 153 breast milk samples analyzed, 20 μL was added to 10 μL of 3M sodium borohydride. An aqueous sodium solution was added, and the mixture was left to stand at room temperature for 30 minutes. Subsequently, 7.5 μL of acetic acid was added, and the mixture was cooled to room temperature. The mixture was left to stand at room temperature for 30 minutes. 15 μL of 1 M ammonium bicarbonate solution and 7.5 μL of 100 mL of ultrapure water was added. Subsequently, 400 mL of 5% sodium lauryl sulfate was added. Add M dithiothreitol solution and heat at 100°C for 10 minutes to denature the protein. After cooling to room temperature, 10 μL of 123 mM iodoacetamide solution was added and the mixture was left to stand at room temperature. 7 μL of 500 mM sodium acetate aqueous solution (pH 6.0), and After adding 9 μL of 10% NP-40 solution, 10 μL of 500 units / μL Pep Add tide-N-glycanase F solution, and let stand at 37°C for 24 hours to form N-crystals. After adding 109 μL of ultrapure water, the total volume was diluted with 10,000 molecular weight cutoff solution. The system was introduced into Vivaspin 500 (Sartorius), and the pressure was 15,000 × g and 25 The mixture was centrifuged at 50°C for 10 minutes, and the permeate was collected. Nitrile was added and subjected to LC / MS analysis. LC / MS analysis was performed as follows. HP The LC was an UltiMate 3000 (Thermo Fisher Scientific Inc.). ), and MS was Q-Exactive (Thermo Fisher Scientific Co., Ltd.). The column used was GlycanPac AXH-1 (Thermo Fisher Scientific). The column temperature was set at 40°C. The mobile phase was acetonitrile. (Solution A) and a 50 mM ammonium formate solution adjusted to pH 4.4 (Solution B) were used. The flow rate was 0.4 mL / min. After the sample was introduced, the ratio of solution B was increased to 10% by 55 minutes. The concentration of solution B was increased linearly from 100 to 35%, and then 35% of solution B was passed through from 55 to 65 minutes. The spray voltage of the SI probe was 3.5 kV, the capillary temperature was 275°C, and the sheath temperature was 100°C. Nitrogen in h gas, auxiliary gas, and collision gas Gas was used. S-Lens RF Level was set to 90. MS spectrum was Full scan measurements were performed in the m / z range of 400-2000 in positive mode. The MS spectra were acquired using Compound Discover 2.1 (Thermophor The N content of milk proteins was analyzed using a previously reported Glycans were detected by comparing them with a glycan database constructed from linked glycans. Compound Discover identifies the extraction chromatograms corresponding to the molecular ion peaks of each glycan. A matogram was created, and the signal area value of each sugar chain detected in each breast milk sample was calculated. The signal area value of the detected glycan was used as the relative quantitative value of each glycan. 16 types of N-linked glycans were detected. The sum of these area values was taken as the relative value for "total N-linked glycans."

[0024] (2-4) Measurement of O-linked glycans The O-linked glycan analysis kit EZGlyco was used for the 153 breast milk samples analyzed. Add the sugar chain release reagent included in the Prep kit (Sumitomo Bakelite Co., Ltd.) and heat at 50°C. After dilution with ultrapure water, the mixture was diluted with Vivaspin (molecular weight cutoff: 10,000). 500, centrifuged at 15,000 x g and 25°C for 15 minutes, and the permeate was collected. 50 μL of acetonitrile was added to 50 μL of the permeate, and the mixture was subjected to LC / MS analysis. The HPLC column was UltiMate 3000 and the MS column was Q-Exactive. The column temperature was set at 40°C. Acetonitrile (Solution A) and 50 mM ammonium formate solution adjusted to pH 4.4 The solution (solution B) was used and passed at a flow rate of 0.4 mL / min. After the sample was introduced, the solution B was The ratio of solution B was increased linearly from 10% to 35%, and then from 55 to 65 minutes, the ratio was increased to 35%. The spray voltage of the ESI probe was 3.5 kV, and the capillary temperature was 275°C. , Sheath gas, auxiliary gas, and collision gas Nitrogen gas was used for as. S-Lens RF Level was set to 90. MS Spectra were measured in the positive mode over a full scan range of m / z 300–2000. The MS spectra were acquired using Compound Discover 2. The O-linked sugars were analyzed using a 1 (Thermo Fisher Scientific) analyzer. The sugar chains were detected by comparing them with the sugar chain database. Extraction chromatograms corresponding to the molecular ion peaks of each sugar chain were created by overlay. The signal area value of each sugar chain detected in the milk sample was calculated. was used as the relative quantitative value of each glycan. Twelve types of O-linked glycans were detected, and their area values were The sum of these values was taken as the relative value for "total O-linked glycans."

[0025] (3) Structural determination of disialo N-linked glycan A and disialo N-linked glycan B (3-1) Structural analysis Regarding one of the 153 samples prepared in the above "Measurement of N-linked glycans," LC / MS / MS measurement was performed to identify disialo N-linked glycan A and disialo N-linked glycan B. The structure of B was analyzed by LC / MS / MS analysis as follows. The iMate 3000 and Q-Exactive MS were used. The column was GlycanP. The column temperature was set at 40°C. The mobile phase was acetonitrile ( Solution A), and 50 mM ammonium formate solution (solution B) adjusted to pH 4.4 The flow rate was 0.4 mL / min. After the sample was introduced, the ratio of solution B was increased from 10% to 55 minutes. The concentration of solution B was increased linearly from 100 to 35%, and then the concentration of solution B was increased to 35% from 55 to 65 minutes. The spray voltage of the I probe was 3.5 kV, the capillary temperature was 275°C, and the sheath temperature was 100°C. Nitrogen gas for gas, auxiliary gas, and collision gas The S-Lens RF Level was set to 90. The MS / MS spectrum was , in positive mode, and data-dependently acquired in Top10 mode. S spectra were analyzed using SimGlycan Software (PREMIER BioSolutions) ft International) and analyzed the disialo N-linked glycan A and The structure of disialo N-linked glycan B was analyzed.

[0026] (3-2) Determination of binding mode In addition, N-acetylneuraminic acid of disialo N-linked glycan A and disialo N-linked glycan B was To determine the bond type of phosphate, sialidase, which specifically degrades α2,3 bonds, or LC after treatment with either sialidase, which cleaves both α2,3 and α2,6 linkages, or For one sample identical to that described in (3-1) above, 5 μL of N-linked glycan solution was analyzed by / MS. 37 μL of ultrapure water and 5 μL of the buffer solution provided with the enzyme were added to the mixture. 3 μL of 1:2 diluted α2,3-Sialidase or Neuraminidase The mixture was added to the PBS and incubated at 37°C for 15 minutes. The enzyme was inactivated by heating in boiling water for 5 minutes. After that, 50 μL of acetonitrile was added and the mixture was subjected to LC / MS analysis. The analysis was carried out as follows: HPLC was performed on an UltiMate 3000 and MS was performed on a Q-Exact The column used was GlycanPac AXH-1, and the column temperature was 40 The mobile phase consisted of acetonitrile (liquid A) and 50 ml of ethanol adjusted to pH 4.4. An aqueous solution of ammonium formate (liquid B) was used, and the flow rate was 0.4 mL / min. After the injection, the proportion of solution B was linearly increased from 10% to 35% by 55 minutes. The ESI probe spray voltage was 3.5 kV, and the capacitance was 1.25 kV. The column temperature was 275°C, and the sheath gas, auxiliary gas, and Nitrogen gas was used as the collision gas. S-Lens RF Level The acquired MS spectra were recorded using Xcalibur software (ThermoF Analysis was performed using a Fischer Scientific Co., Ltd.

[0027] (4) Absolute quantification of disialo N-linked glycans A and B Disialo N-linked glycan A and disialo N-linked glycan B were associated with the incidence of bronchitis. The absolute amount of sugar chain B in breast milk was measured. Content of disialo N-linked sugar chain A in breast milk was measured by the standard addition method using a standard substance of disialo N-linked glycopeptide. The absolute amount of allo-N-linked glycan B in breast milk is calculated by multiplying the absolute amount of disialo-N-linked glycan A by LC / M The area of disialo N-linked glycan B relative to the area of disialo N-linked glycan A obtained from the results of S The relative ratio of the area values was multiplied.

[0028] The method for measuring disialo N-linked glycan A by standard addition method is described below. 20 μL of breast milk was randomly selected from three breast milk samples and 10 μL of 3M hydrogen was added to the sample. An aqueous solution of sodium borohydride was added, and the mixture was left to stand at room temperature for 30 minutes. Acid was added and the mixture was left to stand at room temperature for 30 minutes. 15 μL of 1 M ammonium bicarbonate solution was added. After the addition, 7.5, 5.5, 3.5, or 1.5 μL of ultrapure water was added. (2) Sialylglycopeptide used as a standard substance for glycopeptides with N-linked glycan A Add 0, 2, 4, or 6 μL of an aqueous solution containing 20 μg / mL of EDTA (Tokyo Chemical Industry Co., Ltd.). Subsequently, 5 μL of a 120 mM dithiothreitol aqueous solution was added, and the mixture was incubated at 60°C for 30 minutes. After heating for 1 min, 10 μL of 123 mM iodoacetamide solution was added and the mixture was left at room temperature for 1 min. 20 μL of 40 units / μL trypsin solution was added and incubated at 37°C. The mixture was incubated for 1 hour. Trypsin was inactivated by heating at 100°C for 5 minutes, and then the mixture was Add 370 units / μL of Peptide-N-glycanase F solution. The mixture was left standing at 20°C for 18 hours to release the N-linked glycans. Using the BlotGlyco (Sumitomo Bakelite) chain preparation kit, 2-aminobenzaldehyde was After mide labeling and purification, the product was subjected to LC / MS analysis. The HPLC column was GlycanPac AX. The column temperature was set at 40°C. The mobile phase was acetonitrile (liquid A), A 50 mM ammonium formate solution (solution B) adjusted to pH 4.4 was used, and the flow rate was The solution was passed through at 0.4 mL / min. After the sample was introduced, the ratio of solution B was increased from 10% to 35% within 55 minutes. After linearly increasing the concentration of solution B, 35% solution B was passed from 55 to 65 minutes. The spray voltage was 3.5 kV, the capillary temperature was 275 °C, and the sheath gas was a Nitrogen gas was used as the auxiliary gas and collision gas. The S-Lens RF Level was set to 90. MS measurements were performed in positive mode. Selected ion monitoring measurement was performed at / z 1172.43. The data were analyzed using Xcalibur software. Area values of MS chromatograms By plotting the amount of sialylglycopeptide added against the amount of added sialylglycopeptide, the amount of disialylglycopeptide in breast milk was The amount of N-linked glycan A was absolutely quantified by the standard addition method. Using the relative quantitative values of 153 breast milk samples obtained by "glycan measurement," The absolute quantitative values of disialo N-linked glycan A in each breast milk sample were obtained. The MS chromatogram of disialo N-linked glycan B compared to disialo N-linked glycan A is The absolute quantitative value of disialo N-linked glycan B was obtained from the ratio of the area values.

[0029] (5) Analysis of the relationship between the content of various carbohydrate components in breast milk and bronchitis N-acetylneuraminic acid, oligosaccharides, N-linked glycans, and O The concentrations of disialo N-linked glycans, disialo N-linked glycans A, and disialo N-linked glycans B and the infants' The relationship between the incidence of bronchitis and the risk of bronchitis was analyzed using logistic regression. , whether or not there were older siblings, method of delivery (natural birth, cesarean section), and birth weight were used.

[0030] 2.Results (1) The relationship between the content of various carbohydrate components in breast milk and the incidence of bronchitis in infants The 153 breast milk samples were analyzed for various carbohydrate components (total N-acetylneuraminic acid, total oligosaccharides) The relationship between the content of glycans (total N-linked glycans, total O-linked glycans) and the incidence of infant bronchitis was investigated using a logistic regression analysis. The results showed that the relationship between total N-acetylneuraminic acid concentration and the incidence of bronchitis was A statistically significant relationship was observed between the incidence of serotonin and total oligosaccharides and total N-linked saccharides (Table 1). No such relationship was observed for all N-acetylneuraminic acid chains or all O-linked glycans. The odds ratio between nicotinic acid concentration and the incidence of bronchitis was 0.098, and the regression coefficient was -2.32. Therefore, it was found that infants who drank breast milk with high concentrations of total N-acetylneuraminic acid had a It has been shown that the risk of developing bronchitis is reduced.

[0031] [Table 1]

[0032] Breast milk samples were divided into two groups based on whether the infants had bronchitis, and total N-acetylneuraminic acid in breast milk was analyzed. When comparing laminic acid concentrations, breast milk given by infants who had not suffered from bronchitis had a higher concentration of laminic acid than infants who had suffered from bronchitis. The total N-acetylneuraminic acid concentration was significantly higher in the breast milk than in the milk of the infants who had received the This suggests that infant nutritional formulas may be effective in reducing the risk of bronchitis. , and levels of N-acetylneuraminic acid equivalent to those found in breast milk from infants without bronchitis. This indicates that the infant nutritional composition containing The range of total N-acetylneuraminic acid concentrations in breast milk consumed by infants was 181-977 mg / The total N-acetylneuraminic acid concentration was 395 mg / L (Table 2). The distribution of the concentration of the ions differed from the normal distribution in that it had a wider tail towards higher concentrations. This distribution is also observed in the distribution of height and weight in infants, and is used to create growth curves for infants. In this case, the standard is between the 3rd and 97th percentiles of the overall distribution (2000 Infant and Child Based on this, the total N-acetylneuraminic acid concentration in breast milk was The range of severity was set to the 3rd to 97th percentile. The total N-acetylneuraminic acid concentration in the breast milk consumed by the infants was in the 3rd percentile of the distribution. The lowest value was 221 mg / L. The samples were divided into two groups: those with a content of 246 mg / L or more and those with a content of less than 246 mg / L. The presence or absence of bronchitis was then compared. The correlation was subjected to Fisher's exact test, with p<0.05. N-acetylneuraminic acid levels of 246 mg / L or higher are associated with a low risk of bronchitis. It has been shown that this will happen. Therefore, as a nutritional composition for infants that prevents the onset of bronchitis, it is recommended to add 22 It is necessary to contain 1 mg / L or more of N-acetylneuraminic acid, and preferably 2 mg / L. If the formula concentration is 13%, the solid 10% of the infant nutritional composition should be 46 mg / L or more. It is necessary to compound at least 170 mg of N-acetylneuraminic acid per 100 g of food. It is desirable to incorporate 189 mg or more of N-acetylneuraminic acid. In addition, since the mechanism of bronchitis is the same in infants and adults, this infant nutritional formula The combination of ingredients is thought to be effective for adults and elderly people who are at high risk of bronchitis. Therefore, as a nutritional composition for adults containing the same N-acetylneuraminic acid content as above, It is also effective.

[0033] [Table 2]

[0034] (2) Discovery of N-acetylneuraminic acid compounds that prevent bronchitis in infants The above results suggest that infants who drank breast milk with a high total N-acetylneuraminic acid content had It has been revealed that the risk of developing bronchitis is reduced. Most of them exist as oligosaccharides or sugar chains of glycoproteins, so it is unclear what molecular form they take. We investigated whether N-acetylneuraminic acid in the blood is involved in the prevention of bronchitis.

[0035] Two types of oligosaccharides containing N-acetylneuraminic acid detected in breast milk N-linked glycans (6 types) with cetylneuraminic acid attached, N-acetylneuraminic acid attached The concentrations of the five O-linked glycans synthesized in breast milk and the incidence of infant bronchitis were compared. The association with morbidity was analyzed using logistic regression. Some of the analysis results are shown in Table 3. The analysis revealed that 6'-SL (p=0.035), disialo N-linked glycan A (p=0.03 8), and disialo N-linked glycan B (p=0.046) were associated with the incidence of bronchitis. On the other hand, no such correlation was observed between the content of 3'-SL and the Gender was not recognized. 6'-SL, disialo N-linked glycan A, and disialo N-linked glycan B, and bronchitis Between the incidence and the disease, the regression coefficient was negative and the odds ratio was less than 1. Breast milk with high contents of '-SL, disialo N-linked glycan A, and disialo N-linked glycan B Infants who drank it were shown to have a reduced risk of developing bronchitis.

[0036] [Table 3]

[0037] (3) Confirmation of the preventive effect of 6'-SL on bronchitis in infants Breast milk samples were divided into two groups based on whether the infants had bronchitis, and the 6'-SL concentrations in the breast milk were compared. As a result of the comparison, the breast milk consumed by infants without bronchitis was found to be more effective than the breast milk consumed by infants with bronchitis. The amount of 6'-SL was significantly higher in infant milk than in breast milk (Fig. 2). In order to reduce the risk of bronchitis with a composition, infants who have not had bronchitis should drink it. The results showed that infant nutritional compositions containing 6'-SL at levels equivalent to those in breast milk are effective. The distribution of 6'-SL concentrations in breast milk from infants without bronchitis The range was 25.7 to 701 mg / L, with a median of 203 mg / L (Table 4). Similar to the distribution of N-acetylneuraminic acid, the distribution of 6'-SL concentrations in breast milk is not normally distributed. The distribution was broadened toward the higher concentration side, so the 3rd to 97th percentiles were used as the 6th percentile of breast milk. The 6'-SL concentration range was set as the range of 6'-SL. The level is considered to be 61.0 mg / L or higher, which corresponds to the 3rd percentile or higher. The samples were divided into two groups: those with a 6'-SL concentration of 135 mg / L or more and those with a concentration of less than 135 mg / L. The relationship between the presence and absence of disease was evaluated using Fisher's exact test, with p<0.05. , and a 6'-SL concentration of 135 mg / L or higher in breast milk is associated with a lower risk of bronchitis. It was shown that...

[0038] [Table 4]

[0039] Therefore, as a nutritional composition for infants that prevents the onset of bronchitis, it is recommended to add 61 It is necessary to contain 6'-SL at a concentration of 0.0 mg / L or more, preferably 135 mg / L. If the formula concentration is 13%, the maximum amount per 100g of solids in the infant nutritional composition is It is necessary to incorporate at least 46.9 mg of 6'-SL, preferably 104 mg. It is desirable to combine the above 6'-SL. The mechanism of bronchitis is the same in infants and adults. Therefore, the formulation of this infant nutritional formula is suitable for adults and elderly people who are at high risk of bronchitis. Therefore, it is thought that the same 6'-SL content as above will be effective for adults. It is also effective as a nutritional composition for

[0040] (4) Confirmation of the preventive effect of disialo N-linked glycan A on bronchitis in infants Breast milk samples were divided into two groups based on whether the infants had bronchitis, and the disialo N-linked sugars in breast milk were analyzed. When the concentrations of chain A were compared, breast milk from infants who had not suffered from bronchitis had higher concentrations of chain A than breast milk from infants who had suffered from bronchitis. The amount of disialo N-linked glycan A was significantly higher in the breast milk than in the milk of the infants who had received the serotonin-containing glycan (Fig. 3 This suggests that in order to reduce the risk of bronchitis in infant nutritional formulas, The concentration of disialo N-linked glycan A was equivalent to that of breast milk consumed by infants who were not affected by HIV infection. This indicates that the nutritional composition for infants containing the compound is effective. The range of disialo N-linked glycan A concentrations in breast milk consumed by women was 0.0740–2.8 The median was 0.302 μmol / L (Table 4). Similar to the distribution of phosphoneuraminic acid, the concentration distribution of disialo N-linked glycan A in breast milk was also normally distributed. The distribution was not consistent, and the tail was wider toward higher concentrations. Therefore, the 3rd to 97th percentiles of breast milk were used. The concentration range of disialo N-linked glycan A was set. Breast milk consumed by infants who were not affected by bronchitis. The disialo N-linked glycan A concentration was 0.0947 μmol, which corresponds to the 3rd percentile or higher. The concentration of disialo N-linked glycan A in breast milk is thought to be 0.313 μm The samples were divided into two groups: those with a blood glucose level of 0.5g or higher and those with a blood glucose level of 0.5g or lower. The relationship between the presence or absence of bronchitis was examined using Fis The results of her exact probability test showed p<0.05, and the disialo N-linked When the concentration of sugar chain A is 0.313 μmol / L or higher, the risk of developing bronchitis is reduced. was shown.

[0041] Therefore, as an infant nutritional composition that prevents bronchitis, adding 0. It is necessary that the disialo N-linked glycan A is contained in an amount of 0.947 μmol / L or more, and it is preferable that the disialo N-linked glycan A is contained in an amount of 0.947 μmol / L or more. If the formula concentration is 13%, the infant nutrition A minimum of 0.0728 μmol of disialo N-linked glycan A per 100 g of solids in the composition It is necessary to compound, and preferably 0.241 μmol or more of disialo N-linked sugar It is desirable to combine chain A. The mechanism of bronchitis is the same in infants and adults. Therefore, this infant nutritional formula may also be effective for adults and elderly people who are at high risk of bronchitis. Therefore, it is considered that the adult It is also effective as a nutritional composition for

[0042] (5) Confirmation of the preventive effect of disialo N-linked glycan B on bronchitis in infants Breast milk samples were divided into two groups based on whether the infants had bronchitis, and the disialo N-linked sugars in breast milk were analyzed. When the concentrations of chain B were compared, the breast milk of infants who had not suffered from bronchitis had higher concentrations of chain B than the breast milk of infants who had suffered from bronchitis. The amount of disialo N-linked glycan B was significantly higher in the breast milk than in the milk of the infants who had received the serotonin-containing glycan (Figure 4 This suggests that in order to reduce the risk of bronchitis in infant nutritional formulas, The concentration of disialo N-linked glycan B was equivalent to that of breast milk consumed by infants who were not affected by HIV infection. This indicates that the nutritional composition for infants containing the compound is effective. The range of disialo N-linked glycan B concentrations in breast milk consumed by women was 0.0853-5.0 The median was 0.599 μmol / L (Table 4). Similar to the distribution of phosphoneuraminic acid, the distribution of disialo N-linked glycan B concentrations in breast milk was also normally distributed. The distribution was not consistent, and the tail was wider toward higher concentrations. Therefore, the 3rd to 97th percentiles of breast milk were used. The concentration range was set to disialo N-linked glycan B. Breast milk consumed by infants who were not affected by bronchitis. The disialo N-linked glycan B concentration was 0.122 μmol, which corresponds to the 3rd percentile or higher. / L or more. In addition, the concentration of disialo N-linked glycan B in breast milk is 0.245 μm The samples were divided into two groups: those with a blood glucose level of 0.5g or higher and those with a blood glucose level of 0.5g or lower. The relationship between the presence or absence of bronchitis was examined using Fis Her exact probability test showed that p<0.05 and disialo-N bonds in breast milk were not significant. A concentration of glycoconjugate B of 0.245 μmol / L or higher reduces the risk of developing bronchitis. It was shown that...

[0043] Therefore, as an infant nutritional composition that prevents bronchitis, adding 0. It is necessary that the disialo N-linked glycan B is contained in an amount of 122 μmol / L or more, and preferably If the formula concentration is 13%, the infant nutritional formula Contains at least 0.0935 μmol of disialo N-linked glycan B per 100 g of solid product It is necessary to compound, preferably 0.188 μmol or more of disialo N-linked sugar chains It is desirable to combine B. The mechanism of bronchitis is the same in infants and adults. Therefore, this infant nutritional formula is also effective for adults and elderly people who are at high risk of bronchitis. Therefore, it is thought that the adult-use dietary supplement containing the same amount of disialo N-linked glycan B as above will It is also effective as a nutritional composition for

[0044] (6) Determination of the structures of disialo N-linked glycan A and disialo N-linked glycan B Disialo N-linked glycan A and glycan B in breast milk were found to be associated with the incidence of infant bronchitis. To investigate the structure of disialo N-linked glycan B, α2,3- or α2,6-linked disialo N-linked glycans were When the retention time of the LC / MS chromatogram was compared with that of the standard glycosylated glycan, the disialo N bond was The retention times of disialo N-linked glycan A and disialo N-linked glycan B were both α2,6 disialo N-linked. The results were consistent with those of the standard glycosylation product. In addition, disialo N-linked glycan A and disialo N-linked glycan B were subjected to nonspecific sialidase analysis. When treated with α-acetylglucosamine, degradation of the glycans was observed, whereas α2,3-linked N-acetylglucosamine was observed. When treated with sialidase, which specifically cleaves sialamino acid, the sugar chains were not degraded. Therefore, disialo N-linked glycans A and B are α2,6 disialo N-linked glycans represented by the following structure: It was shown to be a conjugated glycan.

[0045] <Disialo N-linked sugar chain A> Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6)Ma nβ1-4GlcNAcβ1-4GlcNAc <Disialo N-linked sugar chain B> Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6)Ma nβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc

[0046] (7) Confirmation of the preventive effect of α2,6 disialo N-linked glycans on bronchitis in infants The sugar chain structures of disialo N-linked sugar chain A and disialo N-linked sugar chain B are composed of one fucose molecule. The only difference is whether or not they bind to each other. Both have two molecules of N-acetylneuraminic acid at the non-reducing end. The common feature is that laminic acid is linked to galactose by α2,6 bond. N-linked glycans with 2,6 disialo linkages may be associated with the susceptibility to infant bronchitis. Therefore, the distribution of disialo N-linked glycan A and disialo N-linked glycan B in breast milk was investigated. The amount of α2,6 disialo N-linked glycans in breast milk ( The association between bronchitis and pulmonary tuberculosis (molar concentration) was analyzed using logistic regression. The results showed a p=0. 034, and the amount of α2,6 disialo N-linked glycans in breast milk (molar concentration) and bronchial A statistically significant association was found between the incidence of bronchitis and the risk of bronchitis (Table 3). The correlation is due to the negative regression coefficient and the odds ratio being less than 1, which indicates that α2 in breast milk Infants who drank breast milk containing high molar concentrations of 6-disialo N-linked glycans were more likely to develop bronchitis. It has been shown that the risk of developing HIV is reduced.

[0047] Breast milk samples were divided into two groups based on whether the infants had bronchitis, and the α2,6 disialo N The amount of bound sugar chains (molar concentration) was compared, and the infants who did not suffer from bronchitis drank The amount of α2,6 disialo N-linked glycans in breast milk from affected infants was higher than in breast milk from affected infants. (molar concentration) was at a significantly higher level (Figure 5). To reduce the risk of bronchitis, breast milk consumed by infants who did not have bronchitis should be compared with that consumed by infants who did not have bronchitis. It has been shown that infant nutritional compositions containing equivalent levels of α2,6 disialo N-linked glycans are effective. The results show that the α2,6 disialolipids in breast milk of infants who were not affected by bronchitis The range of distribution of N-linked glycan substance amount was 0.179 to 6.76 μmol / L, with a median The distribution of N-acetylneuraminic acid was 0.933 μmol / L (Table 4). The distribution of α2,6 disialo N-linked glycans in breast milk is not a normal distribution, and there is a tail at the higher concentration end. Because of the wide distribution, the 3rd to 97th percentiles were determined as α2,6 disialo N-linked glycans in breast milk. The concentration range of α2,6 disialo N chains was set as the range of the concentration of α2,6 disialo N chains in breast milk ingested by infants who were not affected by bronchitis. Bound glycans are considered to be 0.257 μmol / L or higher, which corresponds to the 3rd percentile or higher. In addition, samples containing α2,6 disialo N-linked glycans in breast milk of 0.440 μmol / L or more The relationship between the presence or absence of bronchitis was examined using Fisher's exact probability test. The results of the evaluation using a constant value of 0.05 indicated that the α2,6 disialo N-linked glycans in breast milk were 0. It was shown that a blood pressure of 440 μmol / L or higher reduces the risk of developing bronchitis.

[0048] Therefore, as an infant nutritional composition that prevents bronchitis, adding 0. It must contain 257 μmol / L or more of α2,6 disialo N-linked glycans. The preferred concentration is 0.440 μmol / L or more. If the formula concentration is 13%, A minimum of 0.198 μmol of α2,6 disialo-N-oligosaccharides per 100 g of solids in the nutritional composition It is necessary to incorporate a glycosylated sugar chain, preferably 0.338 μmol or more of α2,6 disaccharide. It is desirable to incorporate sialo-N-linked glycans. The mechanism of bronchitis differs between infants and adults. The formula for this infant nutritional formula is therefore suitable for adults and children at high risk of bronchitis. Therefore, it is thought that the same α2,6 disialo N-linked sugar as above is effective for elderly people. It is also effective as a nutritional composition for adults containing the chain.

[0049] Example 1: Preparation of N-acetylneuraminic acid-containing infant formula 52.7 kg of whey powder, 239 kg of skim milk, and 28 g of N-acetylneuraminic acid The product was dissolved in 500 kg of water, and 23.9 kg of vegetable oil was mixed with the resulting solution and homogenized. The solution was sterilized, concentrated, and dried using standard methods to obtain 99 kg of powdered milk. 1 kg of the milk component was mixed with the powder to obtain 100 kg of milk powder. The amount of neuraminic acid was 221 mg / 100 g solid.

[0050] [Example 2] Preparation of 6'-sialyllactose-containing infant formula 52.7 kg of whey powder, 239 kg of skim milk, and 50 g of 6'-sialyllactose The product was dissolved in 500 kg of water, and 23.9 kg of vegetable oil was mixed with the resulting solution and homogenized. The solution was sterilized, concentrated, and dried using standard methods to obtain 99 kg of powdered milk. 1 kg of milk component was mixed with powder to obtain 100 kg of milk powder. The amount of lucactose was 50 mg / 100 g solids.

[0051] [Example 3] Preparation of glycopeptide-containing milk powder 52.7 kg of whey powder, 239 kg of skim milk, and 567 mg of sialylglycopeptide The compound was dissolved in 500 kg of water, and 23.9 kg of vegetable oil was mixed with the solution and homogenized. The resulting solution was sterilized, concentrated and dried in the usual way to obtain 99 kg of powdered milk. 1 kg of the alpha-2,6-hydroxybenzoates was added to the milk powder to obtain 100 kg of powdered milk. The disialo N-linked glycan content was 0.198 μmol / 100 g solid.

[0052] [Example 4] Preparation of glycopeptide-containing milk powder 52.7 kg of whey powder, 239 kg of skim milk, and 421 g of egg yolk powder in 500 kg of water The resulting solution was mixed with 23.9 kg of vegetable oil and homogenized. The resulting solution was sterilized and By the usual method, 99 kg of powdered milk was obtained by concentrating and drying. The resulting powdered milk was mixed to obtain 100 kg of formula. The sugar chain was 0.198 μmol / 100 g solid.

[0053] [Example 5] Production of supplements 1g of N-acetylneuraminic acid powder was mixed with 40g of an equal mixture of vitamin C and citric acid. 100g of granulated sugar and 60g of an equal mixture of cornstarch and lactose were added and mixed. The ingredients were packed into stick-shaped bags to produce the supplement for preventing bronchitis of the present invention.

[0054] [Example 6] Production of supplements 10 mg of α2,6 disialo N-linked glycan and 40 g of an equal mixture of vitamin C and citric acid 100 g of granulated sugar and 60 g of a mixture of equal parts of cornstarch and lactose were added and mixed. The mixture was packed into a stick bag to produce a supplement for preventing bronchitis of the present invention.

[0055] Example 7: Production of beverages The ingredients are mixed according to the composition shown in Table 5, filled into a container, and then heat sterilized to produce the gas-containing food product of the present invention. A drink for preventing bronchitis was produced.

[0056] [Table 5]

[0057] Example 8: Production of beverages The ingredients are mixed according to the composition shown in Table 6, filled into a container, and then heat sterilized to produce the gas-containing food product of the present invention. A drink for preventing bronchitis was produced.

[0058] [Table 6] [Industrial Applicability]

[0059] According to the present invention, a nutritional composition containing N-acetylneuraminic acid as an active ingredient can improve the tracheal This ingredient can prevent the onset of bronchitis. It is also effective for elderly people with high blood pressure and those with underlying diseases, so it is suitable for adults and elderly people with high blood pressure. It is also possible to prevent the onset of bronchitis.

Claims

1. A nutritional composition for preventing bronchitis, comprising N-acetylneuraminic acid as an active ingredient.

2. The N-acetylneuraminic acid is any one of a protein, a peptide, an oligosaccharide, and a lipid.

10. The nutritional composition of claim 1, wherein the composition is bound to one or more of the following:

3. The nutrient according to claim 1 or 2, wherein the bond of N-acetylneuraminic acid is an α2,6 bond. composition.

4. A method according to claim 1, wherein the sugar chain containing N-acetylneuraminic acid is present as 6'-sialyllactose.

3. The nutritional composition according to claim 3.

5. The nutrient according to claim 3, wherein the sugar chain containing N-acetylneuraminic acid is present as an N-linked sugar chain. Nutrient composition.

6. 6. The sugar chain containing N-acetylneuraminic acid is an α2,6 disialo N-linked sugar chain. The nutritional composition described in

7. 7. The method according to claim 6, wherein the α2,6 disialo N-linked sugar chain has the following structure (1) or (2): The nutritional composition described above. (1) Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα 1-6)Manβ1-4GlcNAcβ1-4GlcNAc (2) Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα 1-6)Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc

8. 8. The nutritional composition according to any one of claims 1 to 7, which is for infants.

9. A nutritional composition for preventing bronchitis according to any one of claims 1 to 8. Hmm, foods and drinks to prevent bronchitis.

10. A nutritional composition for preventing bronchitis according to any one of claims 1 to 8. A medicine for preventing bronchitis.

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