Nutritional composition for preventing bronchitis

Abstract

To provide a novel nutritional composition by finding ingredients for preventing the onset of bronchitis from breast milk ingredients.SOLUTION: The present invention relates to a nutritional composition for preventing the onset of bronchitis, the nutritional composition containing, as active ingredients, N-acetylneuraminic acid, 6'-sialyllactose, an α2,6 disialo N-linked sugar chain and a disialo N-linked sugar chain of a specific structure. This nutritional composition provides effective ingredients not only for infants but also for aged persons and those having a pre-existing disease at high risk of bronchitis and, therefore, is also efficacious as a nutritional composition that is used in foods and nutritional compositions for adults and aged persons for preventing the onset of bronchitis.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a nutritional composition for preventing bronchitis, and more particularly to a nutritional composition for preventing bronchitis that contains N-acetylneuraminic acid as an active ingredient. [Background technology]

[0002] Bronchitis is a disease caused by a viral or bacterial infection of the airways (bronchi), where the trachea branches off. Bronchitis is most often caused by viral infections such as respiratory syncytial virus, rhinovirus, adenovirus, coronavirus, and influenza virus, but bacterial pathogens such as mycoplasma and Bordetella pertussis can also be a cause. After an initial infection with one of these pathogens, secondary bacterial infections such as pneumococcus and Haemophilus influenzae can cause serious conditions.

[0003] Infants are at high risk of contracting bronchitis because their immune systems are not yet fully developed. In particular, infants up to about three months of age have narrow bronchi that make them less able to expel phlegm, putting them at high risk of developing serious inflammation. Additionally, older people with weakened immune systems are at higher risk of developing bronchitis, which can progress to pneumonia, a serious and life-threatening condition.

[0004] Therefore, preventing bronchitis is important for everyone, from infants to the elderly, in order to lead a healthy social life, and various treatments and preventive methods have been investigated. Regarding treatments for bronchitis, various methods have been developed, including the administration of anti-influenza virus drugs, treatment with various antibiotics, and treatment with antitussives and expectorants. Patent Document 1 discloses an antiulcer agent or a bifidobacterium growth promoter containing a specific sialic acid derivative as an active ingredient. It also demonstrates therapeutic efficacy against infectious diseases caused by viruses such as influenza virus, rotavirus, and herpes virus, but does not demonstrate preventive efficacy against bronchitis. While this patent document demonstrates prevention of diarrhea caused by rotavirus, it does not demonstrate prevention of bronchitis, since rotavirus is not a causative virus of bronchitis. Thus, there are no effective methods for preventing bronchitis, and the only option is to practice regular hand washing and gargling.

[0005] Incidentally, breast milk is known to contain various components that reduce the risk of infectious diseases (Patent Document 2). In particular, carbohydrates, mainly oligosaccharides, contained in breast milk have been reported to inhibit the stage at which certain viruses attach to host cells (Patent Document 3). In searching for active ingredients that prevent the onset of bronchitis, the present inventors focused on the components contained in breast milk. However, it was unclear whether the onset of bronchitis itself could be specifically prevented, and it was not possible to infer what components in breast milk could prevent the onset of bronchitis. [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 believed that if a component that prevents the onset of bronchitis could be found among the components of breast milk, then by incorporating that component into a nutritional composition for infants, it would be possible to provide a nutritional composition for infants that prevents the onset of bronchitis. Furthermore, because the component would be effective not only for infants but also for elderly people and people with underlying diseases who are at high risk of bronchitis, they believed that it would be possible to incorporate the component into foods and nutritional compositions for adults and elderly people and provide them as nutritional compositions that prevent the onset of bronchitis. The object of the present invention is to find components in breast milk that can prevent the onset of bronchitis, and to provide a new nutritional composition. [Means for solving the problem]

[0008] In order to search for components in breast milk that prevent the onset of bronchitis, the inventors statistically analyzed the relationship between the amount of carbohydrate components in breast milk and the infant's history of bronchitis, and conducted extensive research to identify new components that prevent the onset of bronchitis. As a result, they discovered that N-acetylneuraminic acid, 6'-sialyllactose, α2,6 disialo N-linked glycans, and disialo N-linked glycans with specific structures in breast milk are effective in preventing bronchitis, leading to the completion of the present invention. Specifically, the present invention relates to a nutritional composition for preventing the onset of bronchitis, which comprises as active ingredients N-acetylneuraminic acid, 6'-sialyllactose, an α2,6 disialo N-linked glycan, and a disialo N-linked glycan with a specific structure. Specifically, the present invention has the following configuration.

[0009] <1> A nutritional composition for preventing bronchitis, comprising N-acetylneuraminic acid as an active ingredient. <2> The N-acetylneuraminic acid is bound to one or more of a protein, a peptide, an oligosaccharide, or a lipid. <1> The nutritional composition described in <3> The binding mode of N-acetylneuraminic acid is α2,6 bond <1> or <2> The nutritional composition described in <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> The nutritional composition described in <6> The sugar chain containing N-acetylneuraminic acid is an α2,6 disialo N-linked sugar chain <5> The nutritional composition described in <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(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6)Manβ1-4GlcNAcβ1-4GlcNAc (2)Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6)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> A food or drink for preventing bronchitis, comprising the nutritional composition for preventing bronchitis described in any one of the above. <10> <1> from <8> A pharmaceutical product for preventing bronchitis, comprising the nutritional composition for preventing bronchitis according to any one of the above. [Effects of the Invention]

[0010] According to the present invention, a nutritional composition containing N-acetylneuraminic acid as an active ingredient can prevent the onset of bronchitis. Furthermore, since this ingredient is effective not only for infants but also for elderly people and those with underlying diseases who are at high risk of bronchitis, it is possible to prevent the onset of bronchitis in adults and the elderly as well. [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 may be in a free state or bound to other sugars as an oligosaccharide. Furthermore, N-acetylneuraminic acid may be bound to peptides and / or proteins or lipids. Furthermore, N-acetylneuraminic acid may be derived from milk, eggs, bird's nests, or fermentation methods using genetically modified bacteria. It may also be a synthetic, semi-synthetic, or natural product. Natural materials containing natural products may be used as a supply source and incorporated directly into the nutritional composition of the present invention. Here, "natural products" refers to products extracted from natural materials containing N-acetylneuraminic acid by known methods, crudely purified products, or products further purified from such natural materials. Examples include milk-derived glycomacropeptides and sialylglycopeptides, which are obtained by concentrating the sugar chain portion of glycomacropeptides. In the present invention, the bond of N-acetylneuraminic acid is preferably an α2,6 bond. α2,6-linked N-acetylneuraminic acid refers to N-acetylneuraminic acid bound via an α2,6 bond to a galactose residue or an N-acetylgalactosamine residue in an oligosaccharide or glycoprotein sugar chain, and refers to a compound having a Neu5Acα2-6Gal structure or a Neu5Acα2-6GalNAc structure.

[0013] (6'-sialyllactose) In the present invention, the sugar chain containing N-acetylneuraminic acid is preferably present as 6'-sialyllactose (Neu5Acα2-6Galβ1-4Glc). 6'-sialyllactose (hereinafter sometimes simply referred to as 6'-SL) may be derived from milk or from fermentation using genetically modified bacteria, and may be a synthetic, semi-synthetic, or natural product. Furthermore, natural materials containing natural products may be used as a supply source and incorporated directly into the nutritional composition of the present invention. Here, "natural products" refers to those extracted from natural materials containing 6'-sialyllactose by known methods, crudely purified products, or products further purified from these.

[0014] (N-linked glycans) In the present invention, the sugar chain containing N-acetylneuraminic acid is preferably present as an N-linked sugar chain. The N-linked sugar chain containing N-acetylneuraminic acid of the present invention may be in a free state or bound to a peptide and / or protein. It may also be derived from milk, eggs, meat, microorganisms, or fermentation of genetically modified bacteria, and may be a synthetic, semi-synthetic, or natural product. Natural materials containing natural products may also be used as a supply source and incorporated directly into the nutritional composition of the present invention. Here, the term "natural product" refers to a product extracted from a natural material containing an N-linked sugar chain by a known method, a crudely purified product, or a product further purified from such a natural material.

[0015] (α2,6 disialo N-linked glycan) The α2,6 disialo N-linked glycan of the present invention refers to an N-linked glycan that reduces the incidence of bronchitis. The α2,6 disialo N-linked glycan of the present invention may be in a free state or bound to a peptide and / or protein. It may also be derived from milk, eggs, meat, microorganisms, or fermentation methods using genetically modified bacteria, and may be a synthetic, semi-synthetic, or natural product. Natural materials containing natural products may also be used as a supply source and incorporated directly into the nutritional composition of the present invention. Here, the term "natural product" refers to a product extracted from a natural material containing an α2,6 disialo N-linked glycan by a known method, a crudely purified product, or a product further purified from such a natural material.

[0016] Specifically, the α2,6 disialo N-linked glycan in the present invention includes Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6)Manβ1-4GlcNAcβ1-4GlcNAc (hereinafter, sometimes simply referred to as disialo N-linked glycan A) and Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6)Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc (hereinafter, sometimes simply referred to as disialo N-linked glycan B), as well as, for example, Neu5Acα2-6Galβ1-4(Fucα1-3)GlcNAcβ1-2Ma nα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6)Manβ1-4GlcNAcβ1-4GlcNAc, Neu5Acα2-6Galβ1 -4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4(Fucα1-3)GlcNAcβ1-2Manα1-6)Manβ1-4GlcNAcβ1-4 It refers to an N-linked glycan that has two Neu5Acα2,6 bonds at the non-reducing end, such as GlcNAc, Neu5Acα2-6Galβ1-4GlcNAcβ1-2(Neu5Acα2-6Galβ1-4GlcNAcβ1-4)Manα1-3(Galβ1-4GlcNAcβ1-2Manα1-6)Manβ1-4GlcNAcβ1-4GlcNAc.

[0017] The nutritional composition of the present invention must contain an effective amount of N-acetylneuraminic acid to prevent bronchitis, and the preferred range of the amount required per 100 g of solid matter of the nutritional composition is shown below. The N-acetylneuraminic acid content is preferably 170 mg / 100 g solid or more, more preferably 189 mg / 100 g solid or more. If the nutritional composition is powdered milk, the N-acetylneuraminic acid content in the prepared milk is preferably 221 mg / L or more, more preferably 246 mg / L or more. Furthermore, the nutritional composition of the present invention preferably contains 6'-sialyllactose at least 46.9 mg / 100 g solid, more preferably at least 104 mg / 100 g solid, and the 6'-SL content in the prepared milk is preferably at least 61.0 mg / L, more preferably at least 135 mg / L. Furthermore, the nutritional composition of the present invention preferably contains 0.198 μmol / 100 g solid or more of α2,6 disialo N-linked glycans, more preferably 0.338 μmol / 100 g solid or more. Furthermore, the milk after preparation preferably contains 0.257 μmol / L or more of α2,6 disialo N-linked glycans, more preferably 0.440 μmol / L or more. Furthermore, the nutritional composition of the present invention preferably contains 0.0728 μmol / 100 g solid or more, more preferably 0.241 μmol / 100 g solid or more of disialo N-linked glycan A. Furthermore, the content of disialo N-linked glycan A in the prepared milk is preferably 0.0947 μmol / L or more, more preferably 0.313 μmol / L or more. Furthermore, the nutritional composition of the present invention preferably contains 0.0935 μmol / 100 g solid or more, more preferably 0.188 μmol / 100 g solid or more, of disialo N-linked glycan B. Furthermore, the prepared milk preferably contains 0.122 μmol / L or more, more preferably 0.245 μmol / L or more, of disialo N-linked glycan B.

[0018] (nutritional compositions, food and beverages, pharmaceuticals) The nutritional composition for preventing bronchitis of the present invention, which contains N-acetylneuraminic acid or the like as an active ingredient, may be any composition containing an effective amount of N-acetylneuraminic acid or the like, and may contain proteins, fats, carbohydrates, vitamins, minerals, etc. in addition to the active ingredient, and may be used as a nutritional composition for infants and young children, as well as for adults and the elderly. Nutritional compositions for infants and young children include infant formulas and liquid formulas, infant formulas (follow-up milk and growing-up milk), formulas for low birth weight infants, allergen-free foods for those with cow's milk allergies, and lactose-free foods for those with lactose intolerance. Nutritional compositions for adults and the elderly include powdered milk and liquid milk formulas for adults, allergen-free foods for those with milk allergies, lactose-free foods for those with lactose intolerance, and supplements. Furthermore, the food and drink products of the present invention containing N-acetylneuraminic acid etc. for preventing bronchitis may be blended into any food and drink products as long as they contain an effective amount of N-acetylneuraminic acid etc., or may be added to the ingredients during the manufacturing process of the food and drink products. Examples of foods and drinks include, but are not limited to, dairy products such as cheese, fermented milk, lactic acid bacteria drinks, butter, margarine, etc., beverages such as milk drinks, fruit juice drinks, and soft drinks, egg products such as jelly, candy, pudding, and mayonnaise, and sweets and breads such as butter cake. The nutritional composition for preventing bronchitis of the present invention can be used as a raw material for pharmaceuticals as it is, and may be manufactured into tablets, capsules, powders, syrups, etc. by conventional methods. Therefore, when formulating a pharmaceutical containing the composition for preventing bronchitis of the present invention, which contains N-acetylneuraminic acid or the like as an active ingredient, as an active ingredient, it can be formulated by appropriately mixing it with excipients, stabilizers, flavoring agents, etc. that are approved for pharmaceutical use. In addition, the composition containing N-acetylneuraminic acid or the like can be dried as is and used as a powder or dust. Furthermore, it can also be formulated by mixing it with excipients, binders, disintegrants, lubricants, flavoring agents, suspending agents, coating agents, and other optional drugs within a range that does not impair the bronchitis prevention effect. Possible dosage forms include tablets, capsules, granules, powders, dusts, syrups, etc. [Example]

[0019] [Test Example 1] Various carbohydrate components were measured in 153 breast milk samples, and the relationship between the incidence of bronchitis in the infant's first year of life and the carbohydrate components 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 This study used breast milk samples and maternal and infant health questionnaires collected from 1,210 mothers in a prospective cohort study (UMIN ID 000015494) conducted by Snow Brand Beanstalk Co., Ltd. between 2014 and 2019 to investigate the relationship between breast milk components and maternal and infant health status in Japanese women. Furthermore, 200 samples were randomly selected from the 1,210 samples. Breast milk samples collected between one and two months after birth were used for analysis. Parents completed questionnaires investigating their infants' health status six times, every two months, over the course of one year after birth, and the results were collected. From these questionnaires, we tabulated whether or not their infants had been diagnosed with bronchitis by a physician within one year after birth. Cases with unclear diagnosis status were excluded from the study. As a result, 153 samples correctly reported whether or not they had been diagnosed with bronchitis across the six questionnaires over the course of one year, and these were included in the analysis. Of the 153 samples, 18 were from infants diagnosed with bronchitis within one year of birth.

[0021] (2) Measurement of various carbohydrate components in breast milk (2-1) Measurement of total N-acetylneuraminic acid 100 μL of breast milk from 153 samples was mixed with 800 μL of ultrapure water and 100 μL of 1N sulfuric acid and heated to 80°C for 45 minutes. After cooling to room temperature, 1 mL of 0.1N sodium hydroxide solution was added and the mixture was subjected to ultrafiltration with a molecular weight cutoff of 10,000. The permeate was analyzed by HPLC. A DIONEX ICS-5000DP system (Thermo Fisher Scientific Inc.) equipped with an electrochemical detector was used for HPLC analysis, using a CarboPac PA1 column (Thermo Fisher Scientific Inc.). The mobile phase consisted of ultrapure water (Solution A), 200 mM sodium hydroxide solution (Solution B), and 100 mM sodium hydroxide solution containing 600 mM sodium acetate (Solution C) at a flow rate of 1 mL / min. The first 7 minutes were spent at 45% solution B and 10% solution C, after which solution B was linearly decreased to 37.5% by 10 minutes and solution C was linearly increased to 25%. From 10 to 20 minutes, solution B was 37.5% and solution C was 25%. From 20.1 to 25 minutes, solution B was 45% and solution C was 10%. The total amount of N-acetylneuraminic acid in each breast milk sample was calculated from a calibration curve prepared using a standard N-acetylneuraminic acid of known concentration.

[0022] (2-2) Measurement of oligosaccharides 20 μL of breast milk from 153 samples was added to 580 μL of ultrapure water and then filtered through a 10,000 molecular weight cutoff membrane. The permeate was analyzed by HPLC. A DIONEX ICS-5000DP system (Thermo Fisher Scientific Inc.) equipped with an electrochemical detector and a CarboPac PA1 column (Thermo Fisher Scientific Inc.) was used for HPLC analysis. The mobile phase consisted of ultrapure water (Solution A), 200 mM sodium hydroxide solution (Solution B), and 100 mM sodium hydroxide solution containing 600 mM sodium acetate (Solution C) at a flow rate of 1 mL / min. Solution B was run at 50% for the first 10 min, then linearly decreased to 47.5% by 18 min, and solution C was linearly increased to 5%. From 18 to 28 min, the solution B was run at 47.5% and solution C at 5%. From 28 to 32 minutes, the concentration of solution B was linearly decreased to 42%, and then solution C was linearly increased to 16%. From 32 to 39 minutes, 42% solution B and 16% solution C were passed through the tube, and then 30% solution B and 40% solution C were passed through the tube from 39 to 43 minutes. The content of each oligosaccharide in each breast milk sample was calculated from a calibration curve prepared using known concentrations of 2'-fucosyllactose, 3-fucosyllactose, 3'-sialyllactose (hereinafter simply referred to as 3'-SL), 6'-sialyllactose (hereinafter simply referred to as 6'-SL), lactodifucotetraose, lacto-N-tetraose, lacto-N-neotetraose, lacto-N-fucopentaose I, lacto-N-fucopentaose II, lacto-N-difucohexaose I, and lacto-N-difucohexaose II. The sum of these 11 types of oligosaccharides was defined as "total oligosaccharides."

[0023] (2-3) Measurement of N-linked glycans To 20 μL of breast milk from 153 samples analyzed, 10 μL of 3M sodium borohydride solution was added and allowed to stand at room temperature for 30 minutes. 7.5 μL of acetic acid was then added and allowed to stand at room temperature for 30 minutes. 15 μL of 1M ammonium bicarbonate solution and 7.5 μL of ultrapure water were then added. 5 μL of 400 mM dithiothreitol solution containing 5% sodium lauryl sulfate was then added and heated at 100°C for 10 minutes to denature the proteins. After cooling to room temperature, 10 μL of 123 mM iodoacetamide solution was added and allowed to stand at room temperature for 1 hour. 7 μL of 500 mM sodium acetate solution (pH 6.0) and 9 μL of 10% NP-40 solution were added, followed by 10 μL of 500 units / μL Peptide-N-glycanase F solution and allowed to stand at 37°C for 24 hours to release N-linked glycans. After adding 109 μL of ultrapure water, the entire volume was loaded onto a Vivaspin 500 (Sartorius) column with a molecular weight cutoff of 10,000 and centrifuged at 15,000 × g for 10 minutes at 25°C. The permeate was collected. 50 μL of the permeate was mixed with 50 μL of acetonitrile and subjected to LC / MS analysis. LC / MS analysis was performed as follows: HPLC was performed on an UltiMate 3000 (Thermo Fisher Scientific) and MS was performed on a Q-Exactive (Thermo Fisher Scientific). The column temperature was set at 40°C. The mobile phase consisted of acetonitrile (Solution A) and 50 mM ammonium formate solution (Solution B) adjusted to pH 4.4, and the flow rate was 0.4 mL / min. After sample introduction, the proportion of solution B was linearly increased from 10% to 35% by 55 minutes, and then 35% solution B was allowed to flow from 55 minutes to 65 minutes. The ESI probe spray voltage was 3.5 kV, the capillary temperature was 275°C, and nitrogen gas was used for the sheath gas, auxiliary gas, and collision gas. The S-Lens RF Level was set to 90. MS spectra were acquired in the positive mode using full scan measurements in the m / z range of 400-2000.The acquired MS spectra were analyzed using Compound Discover 2.1 (Thermo Fisher Scientific). Glycans were detected by comparing them with a glycan database constructed from previously reported N-linked glycans of milk proteins. In addition, Compound Discover was used to create an extracted chromatogram corresponding to the molecular ion peak of each glycan, and the signal area value of each glycan detected in each breast milk sample was calculated. The calculated signal area value of each glycan was used as the relative quantitative value of each glycan. Sixteen types of N-linked glycans were detected, and the sum of their area values ​​was used as the relative value of "total N-linked glycans."

[0024] (2-4) Measurement of O-linked glycans The 153 breast milk samples analyzed were treated with the glycan release reagent included in the EZGlyco Prep kit (Sumitomo Bakelite Co., Ltd.) O-linked glycan analysis kit and heated at 50°C for 20 minutes. After dilution with ultrapure water, the mixture was loaded onto a Vivaspin 500 column with a molecular weight cutoff of 10,000 and centrifuged at 15,000 × g for 15 minutes at 25°C. The permeate was collected. 50 μL of the permeate was mixed with 50 μL of acetonitrile and subjected to LC / MS analysis. An UltiMate 3000 HPLC column and a Q-Exactive MS column were used. The column temperature was set at 40°C. The mobile phase consisted of acetonitrile (solution A) and 50 mM ammonium formate solution (solution B) adjusted to pH 4.4, and the flow rate was 0.4 mL / min. After sample introduction, the proportion of solution B was linearly increased from 10% to 35% by 55 minutes, and then 35% solution B was passed from 55 minutes to 65 minutes. The ESI probe spray voltage was 3.5 kV, the capillary temperature was 275°C, and nitrogen gas was used for the sheath gas, auxiliary gas, and collision gas. The S-Lens RF level was set to 90. MS spectra were acquired in positive mode using full scan measurements in the m / z range of 300–2000. The acquired MS spectra were analyzed using Compound Discover 2.1 (Thermo Fisher Scientific). Glycans were detected by matching with a database of O-linked glycans. Compound Discover also created extraction chromatograms corresponding to the molecular ion peaks of each glycan, and the signal area values ​​of each glycan detected in each breast milk sample were calculated. The calculated signal area values ​​of each glycan were used as the relative quantification value for each glycan. Twelve types of O-linked glycans were detected, and the sum of their area values ​​was used as a 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 One of the 153 samples prepared in the "Measurement of N-linked Glycans" section above was subjected to LC / MS / MS analysis to analyze the structures of disialo N-linked glycan A and disialo N-linked glycan B. LC / MS / MS analysis was performed as follows: HPLC was performed on an UltiMate 3000 and MS on a Q-Exactive. The column used was a GlycanPac AXH-1, and the column temperature was set at 40°C. The mobile phase consisted of acetonitrile (solution A) and a 50 mM ammonium formate solution (solution B) adjusted to pH 4.4, and the flow rate was 0.4 mL / min. After sample introduction, the proportion of solution B was linearly increased from 10% to 35% by 55 min, and then 35% solution B was used from 55 to 65 min. The ESI probe spray voltage was 3.5 kV, the capillary temperature was 275°C, and nitrogen gas was used for the sheath gas, auxiliary gas, and collision gas. The S-Lens RF Level was set to 90. MS / MS spectra were acquired in positive mode in a data-dependent manner using Top10 mode. The acquired MS / MS spectra were analyzed using SimGlycan Software (PREMIER Biosoft International), and the structures of disialo N-linked glycan A and disialo N-linked glycan B were determined.

[0026] (3-2) Determination of binding mode To determine the N-acetylneuraminic acid linkages in disialo N-glycan A and disialo N-glycan B, we treated them with either a sialidase specifically degrading α2,3-linkages or a sialidase specifically degrading both α2,3- and α2,6-linkages, followed by LC / MS analysis. For the same sample as in (3-1) above, 5 μL of N-glycan solution was mixed with 37 μL of ultrapure water and 5 μL of the buffer solution provided with the enzyme. 3 μL of α2,3-Sialidase or Neuraminidase diluted 10-fold with the buffer solution provided with the enzyme was added and incubated at 37°C for 15 minutes. After heating in boiling water for 5 minutes to inactivate the enzyme, 50 μL of acetonitrile was added and the mixture was subjected to LC / MS analysis. LC / MS analysis was performed as follows: HPLC was performed on an UltiMate 3000 and MS was performed on a Q-Exactive. The column used was a GlycanPac AXH-1, and the column temperature was set to 40°C. The mobile phase consisted of acetonitrile (solution A) and 50 mM ammonium formate aqueous solution (solution B) adjusted to pH 4.4, with a flow rate of 0.4 mL / min. After sample introduction, the proportion of solution B was linearly increased from 10% to 35% by 55 min, and then 35% solution B was used from 55 to 65 min. The ESI probe spray voltage was 3.5 kV, the capillary temperature was 275°C, and nitrogen gas was used as the sheath gas, auxiliary gas, and collision gas. The S-Lens RF Level was set to 90. The acquired MS spectra were analyzed using Xcalibur software (Thermo Fisher Scientific).

[0027] (4) Absolute quantification of disialo N-linked glycans A and B The absolute amounts of disialo N-linked glycan A and disialo N-linked glycan B in breast milk, which have been shown to be associated with bronchitis, were measured. The content of disialo N-linked glycan A in breast milk was measured by the standard addition method using a disialo N-linked glycopeptide standard substance. The absolute amount of disialo N-linked glycan B in breast milk was calculated by multiplying the absolute amount of disialo N-linked glycan A by the relative ratio of the area value of disialo N-linked glycan B to the area value of disialo N-linked glycan A obtained from the LC / MS results.

[0028] The standard addition method for measuring disialo N-linked glycan A is described below. 20 μL of breast milk was randomly selected from 153 breast milk samples and 10 μL of 3 M sodium borohydride solution was added to the sample and allowed to stand at room temperature for 30 minutes. Next, 7.5 μL of acetic acid was added and the sample was allowed to stand at room temperature for 30 minutes. 15 μL of 1 M ammonium bicarbonate solution was added, followed by 7.5, 5.5, 3.5, or 1.5 μL of ultrapure water. Furthermore, 0, 2, 4, or 6 μL of a 20 μg / mL solution of sialylglycopeptide (Tokyo Chemical Industry Co., Ltd.), used as a standard for glycopeptides bearing disialo N-linked glycan A, was added. 5 μL of 120 mM dithiothreitol solution was added and heated at 60°C for 30 minutes. After that, 10 μL of 123 mM iodoacetamide solution was added and the sample was allowed to stand at room temperature for 1 hour. Twenty microliters of a 40-unit / μL trypsin solution was added and the mixture was incubated at 37°C for 1 hour. After inactivating the trypsin by heating at 100°C for 5 minutes, a 500-unit / μL Peptide-N-glycanase F solution was added and the mixture was left at 37°C for 18 hours to release the N-linked glycans. The released N-linked glycans were 2-aminobenzamide-labeled and purified using the BlotGlyco N-linked glycan preparation kit (Sumitomo Bakelite) before being subjected to LC / MS analysis. An UltiMate 3000 HPLC column and a Q-Exactive MS were used. The column temperature was set at 40°C. The mobile phase consisted of acetonitrile (Solution A) and 50 mM ammonium formate solution (Solution B) adjusted to pH 4.4, and the flow rate was 0.4 mL / min. After sample introduction, the proportion of solution B was linearly increased from 10% to 35% by 55 minutes, and then 35% solution B was passed through from 55 minutes to 65 minutes. The ESI probe spray voltage was 3.5 kV, the capillary temperature was 275°C, and nitrogen gas was used for the sheath gas, auxiliary gas, and collision gas. The S-Lens RF Level was set to 90. MS measurements were performed in positive mode using selected ion monitoring of m / z 1172.43. The acquired MS chromatograms were analyzed using Xcalibur software.The amount of disialyl N-linked glycan A in breast milk was absolutely quantified by the standard addition method by plotting the area value of the MS chromatogram against the amount of added sialylglycopeptide. This absolute quantitative value and the relative quantitative values ​​for the 153 breast milk samples obtained by the "Measurement of N-linked glycans" above were used to obtain the absolute quantitative value of disialo N-linked glycan A in each of the 153 breast milk samples. Furthermore, for each breast milk sample, the absolute quantitative value of disialo N-linked glycan B was obtained from the ratio of the area value of the MS chromatogram of disialo N-linked glycan B to that of disialo N-linked glycan A.

[0029] (5) Analysis of the relationship between the content of various carbohydrate components in breast milk and bronchitis Logistic regression was used to analyze the relationship between the concentrations of N-acetylneuraminic acid, oligosaccharides, N-linked glycans, O-linked glycans, disialo N-linked glycan A, and disialo N-linked glycan B in 153 breast milk samples and the incidence of infant bronchitis. Adjustment factors included the infant's sex, presence or absence of siblings, method of delivery (natural birth, cesarean section), and birth weight.

[0030] 2.Results (1) The relationship between the content of various carbohydrate components in breast milk and the incidence of bronchitis in infants Logistic regression analysis was performed on 153 breast milk samples to examine the relationship between the content of various carbohydrate components (total N-acetylneuraminic acid, total oligosaccharides, total N-linked glycans, and total O-linked glycans) and the incidence of infant bronchitis. A statistically significant relationship was observed between total N-acetylneuraminic acid concentration and the incidence of bronchitis (Table 1). However, no such relationship was observed for total oligosaccharides, total N-linked glycans, or total O-linked glycans. The odds ratio and regression coefficient for the relationship between total N-acetylneuraminic acid concentration and the incidence of bronchitis were 0.098 and -2.32, respectively, indicating that infants who drank breast milk with higher total N-acetylneuraminic acid concentrations had a reduced risk of bronchitis.

[0031] [Table 1]

[0032] Breast milk samples were divided into two groups based on whether the infants had bronchitis, and the total N-acetylneuraminic acid concentrations in breast milk were compared. The breast milk from infants without bronchitis had significantly higher levels of total N-acetylneuraminic acid than the breast milk from infants with bronchitis (Figure 1). This suggests that infant nutritional compositions containing N-acetylneuraminic acid at levels equivalent to those in breast milk from infants without bronchitis are effective in reducing the risk of bronchitis. The total N-acetylneuraminic acid concentrations in breast milk from infants without bronchitis ranged from 181 to 977 mg / L, with a median of 395 mg / L (Table 2). The distribution of total N-acetylneuraminic acid concentrations was different from a normal distribution, exhibiting a broader tail toward higher concentrations. This distribution is also observed in the height and weight distributions of infants. When constructing growth curves for infants, the 3rd to 97th percentile of the overall distribution is used as the standard (2000 Infant Physical Development Survey Report). Based on this, we used the 3rd to 97th percentile as the standard range for the total N-acetylneuraminic acid (NNA) concentration in breast milk. The lowest NNA concentration in breast milk consumed by infants without bronchitis was 221 mg / L, corresponding to the 3rd percentile of the distribution. Furthermore, breast milk samples were divided into two groups: those with a NNA content of 246 mg / L or higher and those with a NNA content below 246 mg / L. Fisher's exact test was performed to examine the relationship between the presence and absence of bronchitis, with p<0.05, indicating that a NNA content of 246 mg / L or higher in breast milk is associated with a lower risk of bronchitis. Therefore, for an infant nutritional composition to prevent bronchitis, the formula must contain at least 221 mg / L of N-acetylneuraminic acid, preferably at least 246 mg / L. If the formula concentration is 13%, the infant nutritional composition must contain at least 170 mg of N-acetylneuraminic acid per 100 g of solids, preferably at least 189 mg. Furthermore, because the mechanism of bronchitis is the same in infants and adults, the formulation of this infant nutritional composition is likely to be effective for adults and the elderly who are at high risk of bronchitis. Therefore, it is also effective as a nutritional composition for adults containing the same N-acetylneuraminic acid content as above.

[0033] [Table 2]

[0034] (2) Discovery of N-acetylneuraminic acid compounds that prevent bronchitis in infants These results demonstrate that infants who drink breast milk with a high total N-acetylneuraminic acid content have a reduced risk of developing bronchitis. Because most N-acetylneuraminic acid in breast milk exists as a sugar chain on oligosaccharides and glycoproteins, we explored the molecular form of N-acetylneuraminic acid that is related to the prevention of bronchitis.

[0035] We analyzed the association between the concentrations of two types of oligosaccharides bound to N-acetylneuraminic acid, six types of N-linked glycans bound to N-acetylneuraminic acid, and five types of O-linked glycans bound to N-acetylneuraminic acid detected in breast milk and the incidence of infant bronchitis using logistic regression. Some of the results are shown in Table 3. The analysis revealed a statistically significant association between 6'-SL (p=0.035), disialo N-linked glycan A (p=0.038), and disialo N-linked glycan B (p=0.046) and the incidence of bronchitis, whereas no such association was observed with 3'-SL content. The regression coefficients between 6'-SL, disialo N-linked glycan A, and disialo N-linked glycan B and the incidence of bronchitis were negative, and the odds ratios were less than 1. This indicates that infants who drank breast milk with a high content of 6'-SL, disialo N-linked glycan A, and disialo N-linked glycan B 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. The breast milk from infants without bronchitis had significantly higher levels of 6'-SL than the breast milk from infants with bronchitis (Figure 2). This suggests that infant nutritional compositions containing 6'-SL at levels equivalent to those in breast milk from infants without bronchitis are effective in reducing the risk of bronchitis. The 6'-SL concentrations in the breast milk from infants without bronchitis ranged from 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 was not normal, but rather broadened toward the higher end. Therefore, the 3rd to 97th percentiles were used as the range for 6'-SL concentrations in breast milk. The 6'-SL concentrations in breast milk consumed by infants without bronchitis are considered to be 61.0 mg / L or higher, corresponding to the 3rd percentile or higher. Furthermore, breast milk samples were divided into two groups: those with 6'-SL concentrations of 135 mg / L or higher and those with lower concentrations. The correlation between these two groups and the presence or absence of bronchitis was evaluated using Fisher's exact test. Results showed a p<0.05, indicating that breast milk 6'-SL concentrations of 135 mg / L or higher were associated with a lower risk of bronchitis.

[0038] [Table 4]

[0039] Therefore, for infant nutritional compositions that prevent bronchitis, the formula must contain at least 61.0 mg / L of 6'-SL, preferably at least 135 mg / L. Assuming a 13% formula concentration, the infant nutritional composition must contain at least 46.9 mg of 6'-SL per 100 g of solids, preferably at least 104 mg. Because the mechanism of bronchitis is the same in infants and adults, this infant nutritional composition is likely to be effective for adults and the elderly who are at high risk of bronchitis. Therefore, a nutritional composition containing the same 6'-SL content as above is also effective for adults.

[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 concentrations of disialo N-linked glycan A in the breast milk were compared. The results showed that breast milk from infants without bronchitis had significantly higher levels of disialo N-linked glycan A than breast milk from infants with bronchitis (Figure 3). This indicates that infant nutritional compositions containing disialo N-linked glycan A at concentrations equivalent to those in breast milk from infants without bronchitis are effective in reducing the risk of bronchitis. The range of disialo N-linked glycan A concentrations in breast milk from infants without bronchitis was 0.0740–2.88 μmol / L, with a median of 0.302 μmol / L (Table 4). Similar to the distribution of N-acetylneuraminic acid, the distribution of disialo N-glycan A concentrations in breast milk was not normal, but rather broadened toward higher concentrations. Therefore, the 3rd to 97th percentiles were defined as the range of disialo N-glycan A concentrations in breast milk. The disialo N-glycan A concentrations in breast milk from infants without bronchitis are considered to be 0.0947 μmol / L or higher, which corresponds to the 3rd percentile or higher. Furthermore, breast milk samples were divided into two groups: those with disialo N-glycan A concentrations 0.313 μmol / L or higher and those with lower concentrations. The relationship between breast milk disialo N-glycan A concentrations and the presence or absence of bronchitis was evaluated using Fisher's exact test. Results showed a p<0.05, indicating that breast milk disialo N-glycan A concentrations 0.313 μmol / L or higher were associated with a lower risk of bronchitis.

[0041] Therefore, for infant nutritional compositions that prevent bronchitis, it is necessary for the formula to contain 0.0947 μmol / L or more of disialo N-linked glycan A, preferably 0.313 μmol / L or more. When the formula concentration is 13%, it is necessary to incorporate at least 0.0728 μmol or more of disialo N-linked glycan A per 100 g of solids, preferably 0.241 μmol or more of disialo N-linked glycan A. Because the mechanism of bronchitis is the same in infants and adults, this infant nutritional composition is also considered to be effective for adults and elderly people who are at high risk of bronchitis. Therefore, a nutritional composition for adults containing the same disialo N-linked glycan A content as above is also effective.

[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 concentrations of disialo N-linked glycan B in the breast milk were compared. The results showed that breast milk from infants without bronchitis had significantly higher levels of disialo N-linked glycan B than breast milk from infants with bronchitis (Figure 4). This indicates that infant nutritional compositions containing disialo N-linked glycan B at concentrations equivalent to those in breast milk from infants without bronchitis are effective in reducing the risk of bronchitis. The range of disialo N-linked glycan B concentrations in breast milk from infants without bronchitis was 0.0853–5.06 μmol / L, with a median of 0.599 μmol / L (Table 4). Similar to the distribution of N-acetylneuraminic acid, the distribution of disialo N-glycan B concentrations in breast milk was not normal, but rather broadened toward the higher concentrations. Therefore, the 3rd to 97th percentile was defined as the range of disialo N-glycan B concentrations in breast milk. The disialo N-glycan B concentrations in breast milk consumed by infants who did not suffer from bronchitis are considered to be 0.122 μmol / L or higher, which corresponds to the 3rd percentile or higher. Furthermore, breast milk samples were divided into two groups: those with disialo N-glycan B concentrations 0.245 μmol / L or higher and those with concentrations below 0.245 μmol / L. The correlation between the presence or absence of bronchitis was evaluated using Fisher's exact test. The results were p<0.05, indicating that breast milk disialo N-glycan B concentrations 0.245 μmol / L or higher were associated with a lower risk of bronchitis.

[0043] Therefore, for infant nutritional compositions that prevent bronchitis, it is necessary for the formula to contain 0.122 μmol / L or more of disialo N-linked glycan B, preferably 0.245 μmol / L or more. When the formula concentration is 13%, it is necessary to incorporate at least 0.0935 μmol or more of disialo N-linked glycan B per 100 g of solids, preferably 0.188 μmol or more of disialo N-linked glycan B. Because the mechanism of bronchitis is the same in infants and adults, this infant nutritional composition is also considered to be effective for adults and elderly people who are at high risk of bronchitis. Therefore, a nutritional composition for adults containing the same disialo N-linked glycan B content as above is also effective.

[0044] (6) Determination of the structures of disialo N-linked glycan A and disialo N-linked glycan B To investigate the structures of disialo N-linked glycans A and B in breast milk, which were found to be associated with infant bronchitis, we compared the retention times of LC / MS chromatograms with those of α2,3- or α2,6-linked disialo N-linked glycans. The retention times of both disialo N-linked glycans A and B were consistent with those of α2,6-disialo N-linked glycans. Furthermore, when disialo N-linked glycan A and disialo N-linked glycan B were treated with a nonspecific sialidase, degradation of the glycans was observed, whereas when they were treated with a sialidase that specifically cleaves α2,3-linked N-acetylneuraminic acid, degradation of the glycans was not observed. This indicates that disialo N-linked glycan A and B are α2,6-disialo N-linked glycans represented by the following structure:

[0045] <Disialo N-linked sugar chain A> Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6)Manβ1-4GlcNAcβ1-4GlcNAc <Disialo N-linked sugar chain B> Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6)Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc

[0046] (7) Confirmation of the preventive effect of α2,6 disialo N-linked glycans on bronchitis in infants The only difference between disialo N-linked glycan A and disialo N-linked glycan B is the presence or absence of a single fucose molecule. Both glycans share the common feature of two N-acetylneuraminic acid molecules linked to galactose via α2,6-linkage at the non-reducing end. Therefore, it was suggested that N-linked glycans with this α2,6-disialo linkage might be associated with the incidence of infant bronchitis. Logistic regression was then used to analyze the association between the amount (molar concentration) of α2,6-disialo N-linked glycans in breast milk, which is the sum of the amounts (molar concentrations) of disialo N-linked glycans A and B in breast milk, and bronchitis. The results showed a statistically significant correlation (p = 0.034) between the amount (molar concentration) of α2,6-disialo N-linked glycans in breast milk and the incidence of bronchitis (Table 3). These correlations had negative regression coefficients and odds ratios of less than 1, indicating that infants who drank breast milk with a high molar concentration of α2,6 disialo N-linked glycans had a reduced risk of developing bronchitis.

[0047] Breast milk samples were divided into two groups based on whether the infants had bronchitis, and the molar concentrations of α2,6 disialo N-linked glycans in the breast milk were compared. The results showed that breast milk from infants without bronchitis had significantly higher levels of α2,6 disialo N-linked glycans than breast milk from infants with bronchitis (Figure 5). This suggests that infant nutritional compositions containing α2,6 disialo N-linked glycans at levels equivalent to those found in breast milk from infants without bronchitis are effective in reducing the risk of bronchitis. The range of α2,6 disialo N-linked glycans in breast milk from infants without bronchitis was 0.179–6.76 μmol / L, with a median of 0.933 μmol / L (Table 4). Similar to the distribution of N-acetylneuraminic acid, the distribution of α2,6 disialo N-glycans in breast milk was not normally distributed, but rather broadened toward higher concentrations. Therefore, the 3rd to 97th percentile was used as the concentration range for α2,6 disialo N-glycans in breast milk. The α2,6 disialo N-glycans in breast milk from infants without bronchitis are considered to be ≥0.257 μmol / L, which corresponds to the 3rd percentile. Furthermore, breast milk samples were divided into two groups: those with α2,6 disialo N-glycans ≥0.440 μmol / L and those with ≥0.440 μmol / L. The correlation between these two groups and the presence or absence of bronchitis was evaluated using Fisher's exact test. The results were p<0.05, indicating that breast milk α2,6 disialo N-glycans ≥0.440 μmol / L were associated with a lower risk of bronchitis.

[0048] Therefore, infant nutritional compositions that prevent bronchitis must contain 0.257 μmol / L or more of α2,6 disialo N-linked glycans in the formula after preparation, preferably 0.440 μmol / L or more. Assuming a 13% formula concentration, the infant nutritional composition must contain at least 0.198 μmol or more of α2,6 disialo N-linked glycans per 100 g of solids, preferably 0.338 μmol or more. Because the mechanism of bronchitis is the same in infants and adults, this infant nutritional composition is likely to be effective for adults and elderly people at high risk of bronchitis. Therefore, this infant nutritional composition containing the same α2,6 disialo N-linked glycans as above is also effective as a nutritional composition for adults.

[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 were dissolved in 500 kg of water, and this solution was mixed with 23.9 kg of vegetable oil and homogenized. The resulting solution was sterilized, concentrated, and dried using standard methods to obtain 99 kg of milk powder. The milk powder was then powder-mixed with 1 kg of vitamin and mineral ingredients to obtain 100 kg of milk powder. The amount of N-acetylneuraminic acid in the resulting milk powder was 221 mg / 100 g solids.

[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 were dissolved in 500 kg of water, and this solution was mixed with 23.9 kg of vegetable oil and homogenized. The resulting solution was sterilized, concentrated, and dried using standard methods to obtain 99 kg of milk powder. The milk powder was powder-mixed with 1 kg of vitamin and mineral ingredients to obtain 100 kg of milk powder. The amount of 6'-sialyllactose in the resulting milk powder 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 were dissolved in 500 kg of water, and this solution was mixed with 23.9 kg of vegetable oil and homogenized. The resulting solution was sterilized, concentrated, and dried using standard methods to obtain 99 kg of milk powder. The milk powder was mixed with 1 kg of vitamin and mineral ingredients to obtain 100 kg of final formula. The α2,6 disialo N-linked glycans in the resulting milk powder were 0.198 μmol / 100 g solids.

[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 were dissolved in 500 kg of water, and this solution was mixed with 23.9 kg of vegetable oil and homogenized. The resulting solution was sterilized, concentrated, and dried using standard methods to obtain 99 kg of milk powder. The milk powder was mixed with 1 kg of vitamin and mineral ingredients to finally obtain 100 kg of infant formula. The α2,6 disialo N-linked glycans in the resulting milk powder were 0.198 μmol / 100 g solids.

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

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

[0055] Example 7: Production of beverages The ingredients were mixed according to the composition shown in Table 5, filled into a container, and then heat sterilized to produce a beverage for preventing bronchitis of the present invention.

[0056] [Table 5]

[0057] Example 8: Production of beverages The ingredients were mixed according to the formulation shown in Table 6, filled into a container, and then heat sterilized to produce a beverage for preventing bronchitis of the present invention.

[0058] [Table 6] [Industrial Applicability]

[0059] According to the present invention, a nutritional composition containing N-acetylneuraminic acid as an active ingredient can prevent the onset of bronchitis. Furthermore, since this ingredient is effective not only for infants but also for elderly people and those with underlying diseases who are at high risk of bronchitis, it is possible to prevent the onset of bronchitis in adults and the elderly as well.

Claims

1. A nutritional composition for preventing bronchitis (excluding breast milk), which contains as an active ingredient a sugar chain having the structure shown in (1) or (2) below. (1) Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6)Manβ1-4GlcNAcβ1-4GlcNAc (2) Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6)Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc

2. The nutritional composition for preventing bronchitis according to claim 1, which is for infants.

3. A food or drink for preventing bronchitis, comprising the nutritional composition for preventing bronchitis according to claim 1 or 2.

4. A pharmaceutical for preventing bronchitis, comprising the nutritional composition for preventing bronchitis according to claim 1 or 2.

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