Composition comprising human milk oligosaccharides for use in infants or young children to prevent or treat allergies

Abstract

The present invention relates to a composition comprising human milk oligosaccharides for use in infants or young children for the prevention or treatment of allergies, said composition comprising at least one fucosylated oligosaccharide and at least one N-acetylated oligosaccharide, the invention relates to a composition for preventing and / or treating allergy symptoms in an infant or a young child by increasing propionate production in the body of the infant or the young child.

Description

[0001] This application is a divisional application of the international application filed on January 26, 2017, with application number PCT / EP2017 / 051580 and the invention title "Composition containing human milk oligosaccharides for use in infants or young children to prevent or treat allergic reactions". The international application entered the Chinese national phase on July 5, 2018, with application number 201780005821.6. Technical Field

[0002] This invention relates to nutritional compositions comprising specific oligosaccharides for preventing and / or treating allergic reactions in infants or young children by increasing the production of propionate (especially colonic propionate) in their bodies. The nutritional compositions of this invention are intended to prevent and treat allergic reactions. In a first scenario, the infant or young child is healthy and has a normal risk of developing an allergic reaction or a higher risk due to a first-degree family member having or having previously had an allergic reaction. In a second scenario, the infant or young child has an allergic reaction or is at risk and is therefore ill. Background Technology

[0003] Allergic reactions are among the most common health problems, affecting the lives of people of all ages. Allergic diseases are recognized as an epidemic by the World Health Organization. Studies have shown that the prevalence of allergic reactions has increased over the past few decades. Modern lifestyles, especially urban lifestyles, are associated with a higher prevalence and severity of allergic reactions.

[0004] It has been confirmed that the development of an "allergic phenotype" or "atopic sensitivity" can lead to subsequent sensitization to other allergens and the development of allergic reactions. Therefore, allergic sensitization during childhood, especially early childhood (particularly food allergen sensitization), is dangerous and has attracted considerable attention. Thus, childhood allergic reactions may be the first step in an allergic cascade that can later trigger multiple allergic reactions (a process often referred to as the "atopic March"). For example, it has been shown that children with persistent food hypersensitivity in early childhood have a significantly increased risk of developing allergic rhinitis (hay fever) or asthma later in childhood (Ostblöm et al., 2008). Children with mild food hypersensitivity also have an increased risk of respiratory allergies, but to a lesser extent than children with persistent food hypersensitivity. Therefore, alleviating the severity of food hypersensitivity may be crucial to slowing the "atopic March."

[0005] In this context, managing the onset of allergic reactions and preventing allergic reactions during childhood and infancy is extremely important.

[0006] In the first few years after birth, an infant's immune system is actively developing. Intervening in, preventing, avoiding, managing, mitigating, or modulating allergic reactions in young patients can affect not only their short-term allergic condition but also their long-term allergic condition.

[0007] Evidence suggests that infancy may be a critical period for developing allergies. For various reasons, breastfeeding is recommended for all infants. In particular, breastfeeding has been reported to reduce the risk of allergies in offspring (Lodge, CJ, Breastfeeding and asthma and allergies: a systematic review and meta-analysis, Acta Paediatrica, 2015).

[0008] However, in some cases, due to medical reasons, breastfeeding is insufficient or unsuccessful, or the mother does not choose to breastfeed. Infant formula has been developed for these situations. Fortifiers, which enrich mother's breast milk or infant formula with special ingredients, have also been developed.

[0009] Short-chain fatty acids (SCFAs) are produced, in particular, by the fermentation of dietary fiber in the colon by microorganisms. Propionate, an SCFA, has been shown to protect against allergic inflammation in the lungs and reduce allergic sensitization (the presence of total IgE) (Trompette et al., " Gut Microbiota metabolism of dietary fiber influences allergic airway disease and hematopoiesis (Nature Medicine, 2013). Allergic sensitization (the presence of total IgE) is a marker of increased risk of developing allergic symptoms; therefore, a reduction shown in total IgE should be understood as an indicator of the efficacy of propionate in the prevention and / or treatment of allergic reactions other than allergic inflammation in the lungs, i.e., in general allergy.

[0010] Therefore, increasing propionate levels is a meaningful target for protecting against allergic reactions and allergic symptoms. However, oral SCFAs may be unpalatable.

[0011] Therefore, alternative solutions that are more suitable for infants and young children should be developed.

[0012] All human milk oligosaccharides (HMOs) are the third largest solid component of human milk after lactose and fat. HMOs typically contain lactose at the reducing end and a carbohydrate core at the non-reducing end, which usually contains fucose or sialic acid. More than one hundred HMOs have been isolated and characterized in human milk.

[0013] For various health purposes (primarily immune purposes), several compositions using HMO components (such as fucoidylated oligosaccharides, lactose-N-tetrasaccharides, lactose-N-neotetrasaccharides, and / or sialylated oligosaccharides) have been described.

[0014] However, the use of HMOs to prevent allergic reactions and allergic symptoms has not been adequately studied.

[0015] It is clear that there is a need to develop appropriate methods to prevent and / or treat allergic reactions in infants and young children.

[0016] Moreover, because infants or young children are particularly delicate and not suitable for traditional drug interventions, the way to deliver these health benefits should be a non-drug intervention method that is particularly suitable for young individuals (infants and young children).

[0017] Such health benefits need to be delivered to infants or young children in a way that does not cause side effects and / or is not only easy to deliver but also widely accepted by parents or healthcare professionals.

[0018] Furthermore, the price of delivering this benefit should be fair and reasonable for most people, and affordable for most. Summary of the Invention

[0019] The inventors have discovered that compositions comprising at least one fucosylated oligosaccharide and at least one N-acetylated oligosaccharide can increase the production of colonic propionate in animal models.

[0020] Because propionates are known to be particularly effective against allergic reactions, such compositions can therefore be advantageously used to prevent and / or treat allergic symptoms in infants or young children.

[0021] In a particularly advantageous embodiment, the nutritional composition according to the invention comprises 2'-fucosylated lactose (2-FL) and lactose-N-neotetrasaccharide (LNnT), and particularly the 2FL:LNnT weight ratio is 1:2 to 2:1. Attached Figure Description

[0022] Figure 1 The figure represents the propionate production in the cecum of mice fed a low-fiber diet and a low-fiber diet rich in 5% of different test fibers.

[0023] Abbreviations: Pos ctr = positive control; HMO = human milk oligosaccharide, the weight ratio of 2FL+LNnT tested was 1:1; PDX = polydextrose.

[0024] Figure 2 This represents the ratio of the median SCFA of a fiber-rich diet to the median of a positive control diet.

[0025] Abbreviations: Ctrl pos = positive control; HMO = human milk oligosaccharide, the weight ratio of 2FL+LNnT tested was 1:1; PDX = polydextrose.

[0026] Figure 3 This indicates the IgE levels measured in mice from different groups of mice from the experiments recorded in Example 3.

[0027] Figure 4 This indicates the cecal propionate level measured in mice from different groups of mice from the experiments recorded in Example 3.

[0028] Figure 5 This indicates the propionate level in the feces of mice from different groups of mice from the experiments recorded in Example 3. Detailed Implementation

[0029] As used herein, the following terms have the following meanings.

[0030] The term "infant" refers to a child under 12 months of age.

[0031] The term "preschooler" refers to children aged between one and three years old, also known as toddlers.

[0032] "Cesarean section baby or toddler" refers to a baby or toddler delivered via cesarean section. This means that the baby or toddler was not delivered vaginally.

[0033] "Vaginal delivery infants or toddlers" refers to infants or toddlers delivered vaginally rather than via cesarean section.

[0034] Premature infants are babies or toddlers born before full term. This typically refers to infants or toddlers born before 36 weeks of gestation.

[0035] The term "nutritional composition" refers to a composition that supplies nutrients to an individual. This nutritional composition is typically ingested orally or intravenously. It may include lipid or fat sources, carbohydrate sources, and / or protein sources. In one specific embodiment, the nutritional composition is a ready-to-drink composition, such as a ready-to-drink formula food.

[0036] In one specific embodiment, the composition of the present invention is a hypoallergenic nutritional composition. The term "hypoallergenic nutritional composition" refers to a nutritional composition that is unlikely to cause an allergic reaction.

[0037] In one specific embodiment, the nutritional composition of the present invention is a "synthetic nutritional composition". The term "synthetic nutritional composition" means a mixture obtained by chemical and / or biological means, or a mixture containing components obtained by chemical and / or biological means (including, for example, purification and separation means), which may be chemically identical to a mixture naturally present in mammalian milk, or may contain components identical to those naturally present in mammalian milk (i.e., the synthetic nutritional composition is not breast milk).

[0038] As used herein, the term "infant formula" refers to food intended specifically for the nutrition of infants in the first few months of life, and which meets the diverse nutritional needs of this population (in accordance with Article 2(c) of European Commission Directive 91 / 321 / EEC 2006 / 141 / EC of 22 December 2006 concerning infant formula and Stage 2 infant formula). It also refers to nutritional compositions intended for use in infants, as defined in the Codex Alimentarius Commission (STAN 72-1981) and for infant-specific products (including foods for specific medical purposes). The term "infant formula" encompasses both Stage 1 infant formula and follow-up formula or Stage 2 infant formula.

[0039] "Follow-up formula" or "Stage 2 formula" is offered starting from the 6th month. Infant formula constitutes the main liquid element in the gradually diversifying diet of this group.

[0040] The term "infant food" refers to food designed specifically to provide nutrition for infants or toddlers under one year of age.

[0041] The term "infant cereal composition" refers to food designed specifically for supplying nutrition to infants or young children under one year of age.

[0042] The term "fortifier" refers to a liquid or solid nutritional composition suitable for mixing with breast milk or infant formula.

[0043] The term "weaning period" refers to the period during which breast milk or infant formula is gradually replaced with other foods in the diet of infants or young children.

[0044] The expressions “age in days / weeks / months / age”, “number of days / weeks / months / years after birth”, and “number of days / weeks / months / years after birth” can be used interchangeably.

[0045] The terms “prevention and / or treatment of allergic reactions / allergic responses / allergic symptoms / allergic diseases” refer to the prevention and / or reduction of the frequency and / or incidence and / or severity and / or duration of “allergic reactions” or “allergic responses” or “allergic symptoms” or “allergic diseases.” Incidence is related to the number of “allergic reactions” or “allergic responses” or “allergic symptoms” or “allergic diseases.” Frequency is related to the number of identical “allergic reactions” or “allergic responses” or “allergic symptoms” or “allergic diseases.” This prevention covers reducing the future frequency and / or severity of said “allergic reactions” or “allergic responses” or “allergic symptoms” or “allergic diseases.” The term “future” covers the effects after the intervention ends.

[0046] The terms "later" and "after" are used interchangeably. They refer to effects measured in individuals (infants or toddlers) several weeks, months, or years after birth, such as after 6 months, 8 months, 10 months, 1 year, 2 years, preferably 4 years, more preferably 5 years, even more preferably 7 years, or even older, comparing said effects to the average observations of individuals of the same age. Preferably, it refers to effects observed at least 1 year after birth or at least 2, 5, 7, 10, or 15 years after birth. Thus, the term "later" could refer to observations during infancy, childhood, adolescence, or adulthood. Preferably, it refers to observations during childhood, adolescence, or adulthood. The term "later" also encompasses effects after the intervention has ended.

[0047] The term "health disorder" encompasses any health condition and / or disease and / or functional impairment that affects an individual organism.

[0048] The terms "allergic reaction," "allergic response," "allergic symptoms," or "allergic disease" are used interchangeably. These terms include, but are not limited to, allergic sensitization, food allergy, atopic dermatitis and eczema, asthma, wheezing, allergic rhinitis, rhinoconjunctivitis, eosinophilic esophagitis, hypersensitivity, allergy, and urticaria. Allergic reactions can develop from various allergens, all of which are included within the scope of this invention; non-limiting examples include proteins derived from foods such as milk, eggs, grains, nuts, pollen, animal dander, or house dust mites.

[0049] The term "SCFA" refers to short-chain fatty acids.

[0050] The phrase "increased propionate production" means that, compared to individuals fed a standard composition (i.e., a nutritional composition not containing at least one fucosylated oligosaccharide and at least one N-acetylated oligosaccharide) and / or individuals fed a standard composition supplemented with common fiber (such as polydextrose or pectin), the levels of systemic propionate and / or colonic propionate are higher in individuals fed the nutritional composition according to the invention (i.e., containing at least one fucosylated oligosaccharide and at least one N-acetylated oligosaccharide). Propionate production can be measured using techniques known to those skilled in the art, such as gas-liquid chromatography.

[0051] The statement "increased colonic propionate production" means that, compared to individuals fed a standard composition (i.e., a nutritional composition not containing at least one fucosylated oligosaccharide and at least one N-acetylated oligosaccharide) and / or individuals fed a standard composition supplemented with common fiber (such as polydextrose or pectin), the amount of propionate in the colon (or large intestine) or a portion thereof, such as the cecum, is higher in individuals fed the nutritional composition according to the invention (i.e., containing at least one fucosylated oligosaccharide and at least one N-acetylated oligosaccharide). Propionate production can be measured using techniques known to those skilled in the art, such as gas-liquid chromatography.

[0052] "Breast milk" should be understood as the mother's milk or colostrum.

[0053] The term "HMO" refers to one or more human milk oligosaccharides. These carbohydrates are resistant to enzymatic hydrolysis by digestive enzymes such as the pancreas and / or brush border, indicating that they can exhibit functions not directly related to their calorific value. It has been specifically noted in the art that these carbohydrates play a crucial role in the early development of infants and young children, such as immune system maturation. Many different types of HMOs have been found in human milk. Each individual oligosaccharide is based on a variety of combinations of glucose, galactose, sialic acid (N-acetylneuraminic acid), fucose, and / or N-acetylglucosamine with these molecules, resulting in a large number of diverse oligosaccharides in human milk; more than 130 such structures have been identified to date. Almost all oligosaccharides have a lactose moiety at the reducing end, and the non-reducing end is occupied by sialic acid and / or fucose (if present). HMOs can be acidic (e.g., oligosaccharides containing charged sialic acid) or neutral (e.g., fucoidylated oligosaccharides).

[0054] "Fucosylated oligosaccharides" are oligosaccharides containing fucose residues. These oligosaccharides are neutral. Some examples are 2'-FL (2'-fucosyllactose or 2-fucosyllactose or 2FL or 2-FL), 3-FL (3-fucosyllactose), difucosyllactose, lactose-N-fucopentose (e.g., lactose-N-fucopentose I, lactose-N-fucopentose II, lactose-N-fucopentose III, lactose-N-fucopentose V), lactose-N-fucohexose, lactose-N-difucohexose I, fucosyllactose-N-hexose, fucosyllactose-N-neohexose, difucosyllactose-N-hexose I, difucosyllactose-N-neohexose II, and any combination of these substances.

[0055] The terms “fucosylated oligosaccharides containing 2'-fucosylation epitopes” and “2-fucosylated oligosaccharides” encompass fucosylated oligosaccharides with certain homologous forms. These homologous fucosylated oligosaccharides all contain 2'-fucosylation epitopes, thus suggesting that they have certain homologous functions.

[0056] The term "N-acetylated oligosaccharide" encompasses both "N-acetylaminolactoside" and "oligosaccharide containing N-acetylaminolactoside." This oligosaccharide is a neutral oligosaccharide containing N-acetylaminolactoside residues. Suitable examples are: LNT (lactose-N-tetrasaccharide), para-lactose-N-neohexose (para-LNnH), LNnT (lactose-N-neohexose), or any combination thereof. Other examples are: lactose-N-hexose, lactose-N-neohexose, para-lactose-N-hexose, para-lactose-N-neohexose, lactose-N-octasaccharide, lactose-N-neoctasaccharide, isol-lactose-N-octasaccharide, para-lactose-N-octasaccharide, and lactose-N-decansaccharide.

[0057] The expressions “at least one fucoidylated oligosaccharide” and “at least one N-acetylated oligosaccharide” refer to “at least one type of fucoidylated oligosaccharide” and “at least one type of N-acetylated oligosaccharide”.

[0058] "HMO precursors" are key compounds used to prepare HMOs, such as sialic acid and / or fucose.

[0059] "Sialinated oligosaccharides" are oligosaccharides containing charged sialic acid, that is, oligosaccharides with sialic acid residues. These oligosaccharides are acidic. Some examples are 3-SL (3'-sialyl-lactose) and 6-SL (6'-sialyl-lactose).

[0060] The terms "galacto-oligosaccharide," "galacto-oligosaccharide," and "GOS" are used interchangeably. They refer to oligosaccharides containing two or more galactose molecules that are uncharged and do not contain N-acetyl residues (i.e., they are neutral oligosaccharides). In one specific embodiment, the two or more galactose molecules are linked by β-1,2, β-1,3, β-1,4, or β-1,6 bonds. In another embodiment, "galacto-oligosaccharide" and "GOS" also include oligosaccharides containing one galactose molecule and one glucose molecule linked by β-1,2, β-1,3, or β-1,6 bonds (i.e., a disaccharide).

[0061] The nutritional compositions of the present invention may be in solid form (e.g., powder) or liquid form. The content of various ingredients (e.g., oligosaccharides) may be expressed as g / 100g composition on dry weight when the composition is in solid form (e.g., powder); or as a concentration g / L composition when the composition refers to liquid form (the latter also covers liquid compositions that can be obtained by reconstituted powder with liquids such as milk, water, etc., such as reconstituted infant formula or stage 2 infant formula / follow-up formula or infant cereal products or any other formulations specifically designed for infant nutrition).

[0062] The terms "prebiotic," "fiber(s)," and "fiber(s)" are used interchangeably. They refer to non-digestible carbohydrates that exert a beneficial effect on the host by selectively stimulating the growth and / or activity of healthy bacteria, such as Bifidobacteria in the human colon. Gibson GR, Roberfroid MB. Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. J Nutr. 1995; 125:1401-12 ).

[0063] The term "probiotics" refers to microbial cell preparations or microbial cell components that have beneficial effects on the health or well-being of the host. (Salminen S, Ouwehand A, Benno Y. et al.) Probiotics: how should they be defined (Trends Food Sci. Technol. 1999:10 107-10). Microbial cells are generally bacteria or yeast.

[0064] The term "cfu" should be understood as colony-forming unit.

[0065] Unless otherwise specified, all percentages are by weight.

[0066] Furthermore, in the context of this invention, the terms "comprising" or "including" do not exclude other possible elements. The compositions of this invention (including the various embodiments described herein) may comprise, consist of, or be substantially composed of the following elements: the essential elements and necessary limitations of the invention as described herein, and any other or optional ingredients, components, or limitations as described herein or as required.

[0067] Any references to prior art documents in this specification should not be construed as an admission that such prior art is well-known or constitutes part of common general knowledge in the art.

[0068] The invention will now be described in more detail. It should be noted that the various aspects, features, embodiments, and implementations described herein are compatible and / or can be combined together.

[0069] The present invention therefore relates to a nutritional composition comprising at least one fucoidylated oligosaccharide and at least one N-acetylated oligosaccharide for preventing and / or treating allergic symptoms in infants or young children by increasing propionate production in the body (especially the colon).

[0070] In one embodiment, the invention therefore relates to a nutritional composition comprising at least one fucoidylated oligosaccharide and at least one N-acetylated oligosaccharide for preventing and / or treating allergic symptoms in infants or young children by increasing the production of colonic propionate (especially propionate) in the body of said infants or young children.

[0071] Unbound by theory, the inventors of this invention believe that (one or more) fucosylated oligosaccharides and (one or more) N-acetylated oligosaccharides work synergistically to unexpectedly provide the aforementioned health benefits. This particular combination of oligosaccharides will significantly increase propionate production in an individual and can therefore be used to prevent and / or treat allergic symptoms in infants or young children by increasing propionate (especially colonic propionate) production in the infant or young child.

[0072] The nutritional composition of the present invention comprises at least one fucoidan. One or more types of fucoidan may be present. The fucoidan (one or more) may be selected from a list including: 2'-fucosylvose, 3'-fucosylvose, difucosylvose, lactose-N-fucopentose (such as lactose-N-fucopentose I, lactose-N-fucopentose II, lactose-N-fucopentose III, lactose-N-fucopentose V), lactose-N-fucohexose, lactose-N-difucohexose I, fucoidan-N-fucosylvose V, etc. Hexoses, fucoidyl-lactose-N-neohexoses (such as fucoidyl-lactose-N-neohexose I, fucoidyl-lactose-N-neohexose II), difucosyl-lactose-N-hexose I, difucosyl-lactose-N-neohexose, difucosyl-lactose-N-neohexose I, difucosyl-lactose-N-neohexose II, fucoidyl-para-lactose-N-hexose, trifucosyl-para-lactose-N-hexose I, and any combination thereof.

[0073] In some specific embodiments, the fucoidylated oligosaccharide comprises a 2'-fucosylation epitope. This fucoidylated oligosaccharide may be selected, for example, from a list including: 2'-fucosylation lactose, difucosylation lactose, lactose-N-fucopentose, lactose-N-fucohexose, lactose-N-difucohexose, fucoidyllactose-N-hexose, fucoidyllactose-N-neohexose, difucosylation lactose-N-hexose, difucosylation lactose-N-neohexose, difucosylation lactose-N-neohexose, difucosylation lactose-N-neohexose, fucoidyl-para-lactose-N-hexose, and any combination thereof.

[0074] In a preferred embodiment, the nutritional composition according to the invention comprises 2'-fucosylated lactose (or 2FL, or 2'FL, or 2-FL or 2'-FL). In a specific embodiment, no other type of fucosylated oligosaccharide is present besides 2'-fucosylated lactose; that is, the nutritional composition of the invention comprises only 2'-fucosylated lactose as a fucosylated oligosaccharide.

[0075] Fucosylated oligosaccharides (one or more) can be isolated from natural sources such as animal milk using chromatography or filtration techniques. Alternatively, fucosylated oligosaccharides can be prepared using specialized fucosyltransferases and / or fucosidases via biotechnological means, employing enzyme-based fermentation techniques (recombinant or natural enzymes) or microbial fermentation techniques. In the latter case, the microorganisms can express their natural enzymes and substrates, or can be engineered to produce the corresponding substrates and enzymes. Single microbial cultures and / or mixed cultures can be used. Fucosylated oligosaccharides can be formed starting with acceptor substrates initially having any degree of polymerization (DP), beginning with DP = 1. Alternatively, fucosylated oligosaccharides can be prepared by chemical synthesis from lactose and free fucose. Fucosylated oligosaccharides are also available from, for example, Kyowa Hakko Kogyo Co., Ltd.

[0076] The compositions of the present invention further comprise at least one of N-acetylated oligosaccharides. One or more types of N-acetylated oligosaccharides may be present. One or more N-acetylated oligosaccharides may be, for example, lactose-N-tetrasaccharide (LNT), lactose-N-neotetrasaccharide (LNnT), or any combination thereof. In some embodiments, the N-acetylated oligosaccharide is lactose-N-neotetrasaccharide (LNnT), para-lactose-N-neohexose (para-LNnH), or any combination thereof. In some embodiments, the N-acetylated oligosaccharide is LNnT. In some embodiments, the N-acetylated oligosaccharide is LNT. In some other embodiments, the N-acetylated oligosaccharide is a mixture of LNT and LNnT. In some embodiments, the composition comprises both LNT and LNnT, wherein the LNT:LNnT ratio is 5:1 to 1:2, or 2:1 to 1:1, or 2:1.2 to 2:1.6.

[0077] In a preferred embodiment, the nutritional composition according to the invention comprises lactose-N-neotetrasaccharide (LNnT). In a specific embodiment, no other types of N-acetylated oligosaccharides are contained besides lactose-N-neotetrasaccharide (LNnT), i.e., the nutritional composition of the invention contains only lactose-N-neotetrasaccharide (LNnT) as an N-acetylated oligosaccharide.

[0078] One or more N-acetylated oligosaccharides can be chemically synthesized by enzymatic transfer, i.e., by using glycosyltransferases to transfer sugar units from the donor moiety to the acceptor moiety, as described, for example, in U.S. Patent 5,288,637 and WO 96 / 10086. Alternatively, LNT and LNnT can be prepared by chemically converting free or oligosaccharide-bound ketohexoses (e.g., fructose) into N-acetylated hexosamine or oligosaccharides containing N-acetylated hexosamine, as described in Wrodnigg, TM; Stutz, AE (1999) Angew. Chem. Int. Ed. 38:827-828. The N-acetylated lactoside obtained in this manner can then be transferred to lactose as the acceptor moiety. N-acetylated oligosaccharides can also be produced by biotechnological means based on microbial fermentation technology.

[0079] In a particularly advantageous embodiment of the invention, the nutritional composition comprises 2'-fucosylated lactose (2FL) and lactose-N-neotetrasaccharide (LNnT).

[0080] In another specific embodiment, the nutritional composition of the present invention comprises a mixture of oligosaccharides consisting of 2'-fucosylated lactose (2-FL) and lactose-N-neotetrasaccharide (LNnT). In other words, the nutritional composition of the present invention contains only 2'-fucosylated lactose (2-FL) as a fucosylated oligosaccharide and only lactose-N-neotetrasaccharide (LNnT) as an N-acetylated oligosaccharide.

[0081] In some embodiments, the weight ratio of fucoidylated oligosaccharide to N-acetylated oligosaccharide (e.g., 2FL:LNnT) in the nutritional composition of the present invention is 1:10 to 12:1, such as 1:7 to 10:1 or 1:5 to 5:1, or 2:1 to 5:1 or 1:3 to 3:1, or 1:2 to 2:1, or 1:1 to 3:1, or 1:5 to 1:0.5.

[0082] Before reconstitution with water, the total amount of (one or more) fucoidylated oligosaccharides and (one or more) N-acetylated oligosaccharides present in the nutritional composition of the present invention may be from 0.1% to 10% by weight of the nutritional composition, such as from 0.5% to 7% by weight or from 1% to 5% by weight. For reconstituted ready-to-drink formulations, the target is 0.01% to 1%, more preferably 0.05% to 0.7% or 0.1% to 0.5%.

[0083] The nutritional composition of the present invention may, for example, comprise: - Fucosylated oligosaccharides, in total amounts of 0.2 g / L to 5 g / L, such as 0.5 g / L to 4.5 g / L or 1 g / L to 4 g / L, or in total amounts on dry weight of 0.13 g / 100 g to 3.48 g / 100 g, such as 0.34 g / 100 g to 3.13 g / 100 g or 0.69 g / 100 g to 2.78 g / 100 g; and / or - N-acetylated oligosaccharides, in total amounts of 0.05 g / L composition to 5 g / L composition, for example 0.1 g / L composition to 2 g / L composition or 0.1 g / L composition to 1 g / L composition, or in total amounts on dry weight of 0.03 g / 100 g composition to 3.48 g / 100 g composition, for example 0.07 g / 100 g composition to 1.4 g / 100 g composition or 0.07 g / 100 g composition to 0.7 g / 100 g composition.

[0084] The nutritional composition according to the invention may further comprise at least one additional oligosaccharide (i.e., in addition to (one or more) fucoidylated oligosaccharides and (one or more) N-acetylated oligosaccharides that are essential to be present in the composition) and / or at least one or more fiber and / or at least one or more human milk oligosaccharide precursors. Another oligosaccharide and / or fiber and / or precursor may be selected from a list including: galactooligosaccharides (GOS), fructooligosaccharides (FOS), inulin, xylooligosaccharides (XOS), polydextrose, sialylated oligosaccharides, sialic acid, fucose, and any combination thereof. Their amounts may be from 0% to 10% by weight of the composition.

[0085] In addition to the oligosaccharides contained in the oligosaccharide mixture, suitable commercial products for preparing the nutritional compositions according to the invention include combinations of FOS and inulin, such as those sold by BENEO under the trademark Orafti, or by Tate & Lyle under the trademark STA-LITE. ® Polydextrose for sale.

[0086] In one specific embodiment, the composition according to the invention may comprise sialylated oligosaccharides. One or more sialylated oligosaccharides may be present. The sialylated oligosaccharide(s) may be selected from the group consisting of 3'-sialyl lactose (3-SL), 6'-sialyl lactose (6-SL), and any combination thereof. In some embodiments of the invention, the composition comprises 3-SL and 6-SL. In some specific embodiments, the ratio between 3'-sialyl lactose (3-SL) and 6'-sialyl lactose (6-SL) may be in the range of 5:1 to 1:10, or 3:1 to 1:1, or 1:1 to 1:10. In some specific embodiments, the sialylated oligosaccharide in the composition is 6'-sialyl lactose (6-SL).

[0087] Sialized oligosaccharides (one or more) can be isolated from natural sources such as animal milk using chromatography or filtration techniques. Alternatively, sialylated oligosaccharides can be prepared using specialized sialyltransferases or sialic acid sialidases via biotechnology, through enzyme-based fermentation (recombinant or natural enzymes), chemical synthesis, or microbial fermentation. In the latter case, the microorganisms can express their natural enzymes and substrates, or can be engineered to produce the corresponding substrates and enzymes. Single or mixed microbial cultures can be used. Sialized oligosaccharides can be formed starting with acceptor substrates initially having any degree of polymerization (DP), beginning with DP = 1. Alternatively, sialylated lactose can be prepared via chemical synthesis from lactose and free N'-acetylneuraminic acid (sialic acid). Sialyllated lactose is also commercially available, for example, from KyowaHakko Kogyo in Japan.

[0088] In specific examples, the composition may contain fucoidylated oligosaccharides in total amounts from 0.05 g / L to 5 g / L, such as from 0.1 g / L to 4 g / L or from 0.3 g / L to 2 g / L, or in total amounts on a dry weight basis from 0.03 g / 100 g to 3.5 g / 100 g, such as from 0.1 g / 100 g to 2 g / 100 g or from 0.2 g / 100 g to 1 g / 100 g.

[0089] In one embodiment, the nutritional composition may also comprise at least one BMO (milk oligosaccharide). In another embodiment, the nutritional composition may additionally comprise an oligosaccharide mixture (“BMOS”) comprising 0.1 wt% to 4.0 wt% of one or more N-acetylated oligosaccharides, 92.0 wt% to 99.5 wt% of one or more galactooligosaccharides, and 0.2 wt% to 4.0 wt% of one or more sialylated oligosaccharides. WO2006087391 and WO2012160080 provide some examples of preparing BMO mixtures.

[0090] In some specific embodiments of the invention, the nutritional composition does not contain any (one or more) sialylated oligosaccharides, any GOS and / or any milk oligosaccharides.

[0091] The compositions according to the invention may optionally also contain at least one precursor of human milk oligosaccharides. One or more precursors may be present. For example, human milk oligosaccharide precursors are sialic acid, fucose, or mixtures thereof. In some specific embodiments, the composition contains sialic acid.

[0092] In a specific example, the composition comprises 0 g / L to 3 g / L of human milk oligosaccharide precursor, or 0 g / L to 2 g / L, or 0 g / L to 1 g / L, or 0 g / L to 0.7 g / L, or 0 g / L to 0.5 g / L, or 0 g / L to 0.3 g / L, or 0 g / L to 0.2 g / L of human milk oligosaccharide precursor.

[0093] The compositions according to the invention may comprise 0 g to 2.1 g of one or more human milk oligosaccharide precursors per 100 g composition on a dry weight basis, for example, 0 g to 1.5 g, 0 g to 0.8 g, or 0 g to 0.15 g of one or more human milk oligosaccharide precursors per 100 g composition on a dry weight basis.

[0094] The nutritional composition of the present invention may also contain at least one probiotic (or probiotic strain), such as a probiotic strain.

[0095] The most commonly used probiotics are mainly bacteria and yeasts belonging to the following genera: Lactobacillus species ( Lactobacillus spp. Streptococcus species ( Streptococcus spp. ), Enterococcus spp., Bifidobacterium spp. Bifidobacterium spp. ) and yeast species ( Saccharomyces spp. ).

[0096] In some specific implementations, the probiotics are probiotic bacterial strains. In some specific implementations, they are specifically Bifidobacterium (Bifidobacterium). Bifidobacteria) and / or lactobacillus ( Lactobacilli ).

[0097] Suitable probiotic strains include Lactobacillus rhamnosus, trademarked LGG, from Valio Oy, Finland. Lactobacillus rhamnosus ATCC 53103, Lactobacillus rhamnosus CGMCC 1.3724, Lactobacillus casei ( Lactobacillus paracasei CNCM I-2116, Lactobacillus johnsonii ( Lactobacillus johnsonii CNCM I-1225, BLIS Technologies Limited, New Zealand, sells Streptococcus salivarius under the trade name KI2. Streptococcus salivarius DSM 13084, Bifidobacterium lactis specially sold by Christian Hansen company, Denmark under the trademark Bb12; CNCM 1-3446, Bifidobacterium longum sold by Morinaga Milk Industry Co. Ltd., Japan under the trademark BB536. Bifidobacterium longum ATCC BAA-999, and Bifidobacterium breve sold by Danisco under the trademark Bb-03. Bifidobacterium breve Bifidobacterium breve, sold by Morinaga under the trademark M-16V, and Bifidobacterium infantis, sold by Procter & Gamble Co. under the trademark Bifantis. Bifidobacterium infantis ), and Bifidobacterium breve sold by the Rosell Institute of Biology in Canada under the trademark R0070.

[0098] The nutritional composition according to the invention may contain 10e3 to 10e12 cfu of probiotic strains / g composition on a dry weight basis, more preferably 10e7 to 10e12 cfu of probiotic strains / g composition, such as 10e8 to 10e10 cfu of probiotic strains / g composition.

[0099] In one embodiment, the probiotics are live. In another embodiment, the probiotics are non-replicating or inactivated. In some other embodiments, both live and inactivated probiotics may be present simultaneously.

[0100] The nutritional compositions of the present invention may further comprise at least one bacteriophage (bacterial bacteriophage) or a mixture of bacteriophages, which are preferably targeted at pathogenic streptococci, Haemophilus influenzae (Hib), etc. HaemophilusMoraxella ( ) Moraxella ) and Staphylococcus ( Staphylococci ).

[0101] The nutritional compositions according to the present invention may be, for example, infant formula, stage 1 infant formula, stage 2 infant formula, or follow-up formula, baby food, infant cereal composition, fortifier (such as human milk fortifier), or supplement. In some specific embodiments, the compositions of the present invention are infant formula, fortifiers, or supplements intended for use in infants aged 4 months or 6 months. In a preferred embodiment, the nutritional compositions of the present invention are infant formula.

[0102] In some other embodiments, the nutritional composition of the present invention is a fortifier. The fortifier may be a breast milk fortifier (e.g., human milk fortifier) ​​or a formula food fortifier (such as an infant formula fortifier or a stage 2 / follow-up formula fortifier).

[0103] When a nutritional composition is a supplement, it can be provided in unit dose form.

[0104] The nutritional compositions of the present invention may be in solid (e.g., powder), liquid or gel form.

[0105] The nutritional compositions according to the invention typically contain a protein source. The amount of protein can be from 1.5 g / 100 kcal to 3 g / 100 kcal. In some embodiments, particularly when the composition is intended for preterm infants, the amount of protein can be from 2.4 g / 100 kcal to 4 g / 100 kcal or higher than 3.6 g / 100 kcal. In some other embodiments, the amount of protein can be less than 2.0 g / 100 kcal, for example from 1.8 g / 100 kcal to 2 g / 100 kcal, or less than 1.8 g / 100 kcal.

[0106] The type of protein is considered irrelevant to this invention, provided that the minimum requirements for essential amino acid content are met and satisfactory growth is ensured. Therefore, protein sources based on whey, casein, and mixtures thereof, as well as soy-based protein sources, can be used. Regarding the whey protein of interest, the protein source can be based on acidic whey, sweet whey, or mixtures thereof, and can contain any desired proportions of α-lactalbumin and β-lactoglobulin.

[0107] In some advantageous implementations, the protein source is whey-based (i.e., more than 50% of the protein comes from whey protein, such as 60% or 70%).

[0108] The protein can be a whole protein, a hydrolyzed protein, or a mixture of both. The term "whole" means that the major components of the protein are intact, i.e., the molecular structure is unchanged, for example, at least 80% of the protein is unchanged, such as at least 85% of the protein is unchanged, preferably at least 90% of the protein is unchanged, and even more preferably, at least 95% of the protein is unchanged, such as at least 98% of the protein is unchanged. In one specific embodiment, 100% of the protein is unchanged.

[0109] The term "hydrolyzed" means, in the context of this invention, that the protein has been hydrolyzed or broken down into its constituent amino acids.

[0110] The protein can be completely or partially hydrolyzed. For example, for infants or young children considered at risk of developing bovine milk allergies, providing partially hydrolyzed protein (2% to 20% hydrolysis) may be preferable. If a hydrolyzed protein is required, the hydrolysis process can be carried out as needed and as is known in the art. For example, whey protein hydrolysates can be prepared by enzymatic hydrolysis of whey fractions in one or more steps. If the whey fraction used as a raw material is substantially lactose-free, it has been found that the protein undergoes much less lysine blocking during hydrolysis. This allows the degree of lysine blocking to be reduced from about 15% by weight of total lysine to less than about 10% by weight of lysine; for example, about 7% by weight of lysine, which greatly improves the nutritional quality of the protein source.

[0111] In one embodiment of the invention, at least 70% of the protein is hydrolyzed, preferably at least 80%, such as at least 85%, and even more preferably at least 90%, such as at least 95%, particularly at least 98%. In one specific embodiment, 100% of the protein is hydrolyzed.

[0112] In one specific embodiment, the protein in the nutritional composition is hydrolyzed, fully hydrolyzed, or partially hydrolyzed. The degree of hydrolysis (DH) of the protein can be 8 to 40, or 20 to 60, or 20 to 80, or greater than 10, 20, 40, 60, 80, or 90.

[0113] In one embodiment, the nutritional composition according to the invention is a hypoallergenic composition. In another embodiment, the composition according to the invention is a hypoallergenic nutritional composition.

[0114] The nutritional compositions according to the invention typically contain a carbohydrate source. This is particularly preferred when the nutritional compositions of the invention are for infant formula. In this case, any carbohydrate source commonly found in infant formula can be used, such as lactose, sucrose, saccharin, maltodextrin, starch, and mixtures thereof, but one of the preferred carbohydrate sources is lactose.

[0115] The nutritional compositions according to the invention typically contain a lipid source. This is particularly relevant when the nutritional compositions of the invention are for infant formula. In this case, the lipid source can be any lipid or fat suitable for use in infant formula. Some suitable fat sources include palm oil, high-oleic sunflower oil, and high-oleic safflower oil. The essential fatty acids linoleic acid and α-linolenic acid may also be added, as well as small amounts of oils containing large amounts of pre-formed arachidonic acid and docosahexaenoic acid, such as fish oil or microbial oil. The ratio of n-6 fatty acids to n-3 fatty acids in the fat source can be from about 5:1 to about 15:1, for example from about 8:1 to about 10:1.

[0116] The nutritional compositions of the present invention may also contain all vitamins and minerals considered essential for a daily diet, present in significant amounts in the composition. Minimum requirements for certain vitamins and minerals have been determined. Examples of minerals, vitamins, and other nutrients optionally present in the compositions of the present invention include vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin E, vitamin K, vitamin C, vitamin D, folic acid, inositol, niacin, biotin, pantothenic acid, choline, calcium, phosphorus, iodine, iron, magnesium, copper, zinc, manganese, chlorine, potassium, sodium, selenium, chromium, molybdenum, taurine, and L-carnitine. Minerals are typically added in the form of salts. The presence and amounts of specific minerals and other vitamins will vary depending on the target population.

[0117] If necessary, the nutritional compositions of the present invention may contain emulsifiers and stabilizers, such as soybean, lecithin, monoglyceride citrate, and diglyceride citrate.

[0118] The nutritional compositions of the present invention may also contain other substances that may have beneficial effects, such as lactoferrin, nucleotides, nucleosides, etc.

[0119] The nutritional compositions of the present invention may also contain one or more carotenoids. In some specific embodiments of the present invention, the nutritional compositions of the present invention do not contain any carotenoids.

[0120] The nutritional compositions according to the invention can be prepared by any suitable means. The compositions will now be described by way of example.

[0121] For example, formula foods such as infant formula can be prepared by blending protein sources, carbohydrate sources, and fat sources together in appropriate proportions. If an emulsifier is used, it can be added at this stage. Vitamins and minerals can be added at this stage, but they are usually added later to avoid thermal degradation. Before blending, any lipophilic vitamins, emulsifiers, etc., can be dissolved in the fat source. Water (preferably water that has undergone reverse osmosis) can then be added to form a liquid mixture. A suitable water temperature is in the range of about 50°C to about 80°C to aid in the dispersion of the components. Commercially available liquefying agents can be used to form the liquid mixture.

[0122] Especially if the final product is in liquid form, one or more fucoidylated oligosaccharides and one or more N-acetylated oligosaccharides can be added at this stage. If the final product is a powder, these components can also be added at this stage as needed.

[0123] Then, the liquid mixture is homogenized in, for example, in two stages.

[0124] The liquid mixture can then be heat-treated to reduce the bacterial load, for example by rapidly heating the liquid mixture to a temperature in the range of about 80°C to about 150°C and holding it for a duration of about 5 seconds to about 5 minutes. This can be done by steam injection, autoclaving, or a heat exchanger (e.g., a plate heat exchanger).

[0125] The liquid mixture is then cooled, for example, to about 60°C to about 85°C by rapid cooling. It is then homogenized again, for example, in two stages, where the pressure in the first stage is about 10 MPa to about 30 MPa, and the pressure in the second stage is about 2 MPa to about 10 MPa. The homogenized mixture can then be further cooled to add any heat-sensitive components, such as vitamins and minerals. The pH and solids content of the homogenized mixture can then be conveniently adjusted.

[0126] If the final product is to be a powder, the homogenized mixture is transferred to a suitable drying apparatus, such as a spray dryer or freeze dryer, and then converted into a powder. The moisture content of the powder should be less than about 5% by weight. Alternatively, one or more fucoidylated oligosaccharides and one or more N-acetylated oligosaccharides may be added at this stage by dry mixing them with one or more probiotic strains (if used), or by blending them with one or more probiotic strains in the form of crystalline syrup, followed by spray drying or freeze drying of the mixture.

[0127] If a liquid composition is preferred, the homogenized mixture can be sterilized and then packaged into a suitable container under aseptic conditions, or it can be packaged into a container first and then sterilized.

[0128] In another embodiment, the composition of the present invention may be a supplement.

[0129] Supplements may be in the form of tablets, capsules, lozenges, or liquids. Supplements may also contain protective hydrocolloids (such as gums, proteins, modified starches), binders, film-forming agents, encapsulation agents / materials, wall / shell materials, matrix compounds, coatings, emulsifiers, surfactants, solubilizers (oils, fats, waxes, lecithin, etc.), adsorbents, carriers, fillers, co-compounds, dispersants, wetting agents, processing aids (solvents), flow agents, flavor masking agents, weighting agents, gelling agents, and gelling agents. Supplements may also contain conventional pharmaceutical additives and adjuvants, excipients, and diluents, including but not limited to: water, gelatin of any origin, plant gums, lignin sulfonates, talc, sugars, starches, gum arabic, vegetable oils, polyalkylene glycols, flavoring agents, preservatives, stabilizers, emulsifiers, buffers, lubricants, coloring agents, wetting agents, fillers, etc.

[0130] In addition, supplements may contain organic or inorganic carrier materials suitable for oral or parenteral administration, as well as vitamins, minerals, trace elements, and other micronutrients recommended by government agencies such as the USRDA.

[0131] The nutritional composition according to the present invention is for use with infants or young children. The infants or young children may be full-term or premature. In one specific embodiment, the nutritional composition of the present invention is for premature infants or young children.

[0132] The nutritional composition of the present invention can also be used for infants or young children delivered by cesarean section or vaginal delivery.

[0133] In some embodiments, the nutritional compositions according to the invention can be used before and / or during the weaning period.

[0134] In some embodiments, the nutritional compositions according to the invention are used for infants or young children at risk of developing allergies. In some embodiments, the nutritional compositions according to the invention are used for infants or young children born to mothers who have allergies. Indeed, scientific evidence continues to suggest that infants born to mothers who have allergies have a greater risk of developing allergies later in life than infants born to mothers who do not have allergies.

[0135] The age and duration of administration (providing or feeding) of the nutritional composition can be determined based on availability and need.

[0136] Because the nutritional composition is also used for preventative purposes (prevention of future health problems), it can be provided, for example, immediately after birth. The compositions of the present invention can also be provided within one week, two weeks, three weeks, one month, two months, three months, four months, six months, eight months, ten months, one year, two years, or even longer after birth. In some particularly advantageous embodiments of the invention, the nutritional composition is provided (or administered) to the infant during the first four or six months after birth.

[0137] In some other embodiments, the nutritional composition of the present invention is provided a few days (e.g., 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 20 days…), a few weeks (e.g., 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks…), or a few months (e.g., 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months…) after birth. This may specifically refer to cases where the infant is premature, but it is not necessary.

[0138] In one embodiment, the composition of the present invention is provided to an infant or young child as a supplementary composition to breast milk. In some embodiments, the infant or young child receives breast milk for at least the first 2 weeks, the first 1 month, 2 months, 4 months, or 6 months. In one embodiment, the nutritional composition of the present invention is provided to the infant or young child after this period of breast milk provision, or provided to the infant or young child together with breast milk during this period of breast milk provision. In another embodiment, the composition is provided to the infant or young child as the sole or primary nutritional composition for at least a period of time (e.g., after at least 1 month, 2 months, or 4 months), for at least 1 month, 2 months, 4 months, or 6 months.

[0139] In one embodiment, the nutritional composition of the present invention is a complete nutritional composition (meeting all or most of an individual's nutritional needs). In another embodiment, the nutritional composition is a supplement or fortifier intended for use, for example, to supplement human milk or to supplement infant formula or stage 2 infant formula.

[0140] The inventors have discovered that specific HMO interventions in animal models significantly increase propionate production in the cecum (part of the colon).

[0141] As mentioned in the background section, propionate is known to protect against allergic reactions.

[0142] The nutritional composition according to the invention can therefore be used to prevent and / or treat allergic symptoms in infants or young children by increasing propionate production (especially colonic propionate production) in the infant's or young child's body.

[0143] The target health benefits of this invention can be achieved using a nutritional composition that increases the production of propionate in the colon, particularly in the cecum, of the infant or young child. In one specific embodiment, propionate production is determined by gas-liquid chromatography and can be expressed as nmol / mg dry weight.

[0144] In one specific embodiment, the colonic propionate yield is increased by at least 10%, or at least 15%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60% compared to the colonic propionate yield obtained using a nutrient composition that does not contain at least one fucoidylated oligosaccharide and at least one N-acetylated oligosaccharide.

[0145] In one specific embodiment, colonic propionate production is increased by at least 10%, or at least 15%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70% compared to colonic propionate production obtained using a nutritional composition supplemented with common fiber (such as polydextrose or pectin).

[0146] This indicates a new clinical situation in which the prevention of allergic symptoms can be achieved in a new way.

[0147] Other purposes: Another object of the present invention is to prepare a nutritional composition for preventing and / or treating allergic symptoms in infants or young children by using at least one fucoidylated oligosaccharide and at least one N-acetylated oligosaccharide.

[0148] Another object of the present invention is a pharmaceutical composition comprising at least one fucoidylated oligosaccharide and at least one N-acetylated oligosaccharide, the pharmaceutical composition being used to prevent and / or treat allergic symptoms in an infant or young child by increasing propionate production (especially colonic propionate production) in the infant or young child.

[0149] Another object of the present invention is to use at least one fucoidylated oligosaccharide and at least one N-acetylated oligosaccharide to prevent and / or treat allergic symptoms in infants or young children by increasing propionate production (especially colonic propionate production) in the body of the infant or young child.

[0150] Another object of the present invention relates to a method for preventing and / or treating allergic symptoms in an infant or young child by increasing propionate production (especially colonic propionate production) in the infant or young child, the method comprising administering to the infant or young child a nutritional composition comprising at least one fucosylated oligosaccharide and at least one N-acetylated oligosaccharide.

[0151] The different implementation schemes, details, and examples described above (e.g., concerning the type and content of oligosaccharides, nutritional compositions, application, target populations, etc.) are also applicable to all these other purposes.

[0152] Example

[0153] The following examples illustrate some specific embodiments of the compositions used according to the invention. These examples are given for illustrative purposes only and should not be construed as limiting the invention, as various changes can be made thereto without departing from the spirit of the invention.

[0154] Example 1

[0155] Table 1 below provides examples of the composition of nutritional compositions (e.g., infant formula) according to the present invention. These compositions are given by way of example only.

[0156]

[0157] Table 1: Examples of the composition of nutritional compositions (e.g., infant formula) according to the present invention.

[0158] Example 2

[0159] Research Description

[0160] Five-week-old female BALB / cByJ CRL mice from the Charles River were divided into several groups and fed for 6 weeks according to the following protocol: - Week 1: All groups were fed a low-fiber diet (composition detailed in Table 2). - Weeks 2 to 6: Control group (Group A): Low-fiber diet (same as week 1) Test groups (Groups B to D): Low-fiber diet (same as week 1), supplemented with 5% test fiber by weight (5% of total low-fiber diet replaced by 5% test fiber).

[0161] Table 2: Composition of a low-fiber diet

[0162] The following fibers were tested: HMO = Human milk oligosaccharides. The test was conducted with a weight ratio of 1:1 for 2FL+LNnT.

[0163] PDX = Polydextrose

[0164] pectin

[0165] Table 3 provides a summary of the different test groups and diets.

[0166]

[0167] Table 3: Test group and diet in this study

[0168] Six weeks later, animals in each group were sacrificed, and the contents of the cecum were collected. SCFA production was measured by gas-liquid chromatography (GLC; amount of SCFA in nmol / mg dry weight). The following SCFAs were measured: propionate, butyrate, valerate, and acetate.

[0169] Measurements were performed using the following protocol: SCFA in an acidic solution (pH 2.0 to 3.0) was separated on a GLC column coated with a polar stationary phase. This allows for minimal sample preparation (no derivatization) and simple, basic FID detection. Any residual bacterial activity was inactivated using an acidic phosphate buffer containing HgCl2, and SCFA was extracted from the cecum using an internal standard (2,2-dimethylbutyric acid) for GLC analysis. After centrifugation, the aseptically filtered supernatant was prepared for GLC analysis. SCFA was measured simultaneously.

[0170] The median ratio was calculated to compare the effects of diets rich in different fibers on SCFA production.

[0171] Discover

[0172] Diets rich in HMOs significantly increase propionate production (see [link to diet]). Figure 1Compared to the positive control, its yield increased by approximately 69%. Compared to pectin and PDX, its yield increased by 73% and 75%, respectively. This is quite surprising, as pectin is generally considered a high inducer of SCFA (Stark et al., J Nutr. 1993, In vitro production of short-chain fatty acids by bacterial fermentation of dietary fiber compared with effects of those fibers on hepatic sterol synthesis in rats; Yang et al., Anaerobe, 2013, In vitro characterization of the impact of selected dietary fibers on fecal microbiota composition and short chain fatty acid production).

[0173] Figure 2 This represents the ratio of the median SCFA for each tested food group in each fiber-rich diet to the median SCFA in the positive control diet (i.e., the low-fiber diet only). A ratio of 1 (black line) indicates no difference between the fiber-rich and control diets. A ratio below 1 indicates that the corresponding SCFA is higher in the control diet compared to the fiber-rich diet, while a ratio above 1 indicates that the corresponding SCFA is higher in the fiber-rich diet than in the control diet.

[0174] Diets rich in PDX and pectin induced less release of various SCFAs. Conversely, diets rich in HMOs induced more propionate release than low-fiber diets. The HMO-rich diet was the only one that promoted propionate release, showing a very significant difference compared to other types of SCFAs and other tested fibers.

[0175] The inventors were therefore surprised to find that mice fed with a composition comprising at least one fucosylated oligosaccharide and at least one N-acetylated oligosaccharide had significantly higher cecal (and therefore colonic) propionate production.

[0176] Due to the known properties of propionate, particularly in the prevention and treatment of allergic reactions, compositions comprising at least one fucosylated oligosaccharide and at least one N-acetylated oligosaccharide will therefore be effective for the prevention and / or treatment of allergic symptoms in infants or young children.

[0177] Example 3

[0178] method : Female Balb / c mice aged 5 to 8 weeks were given 50-200 µL of Aspergillus fungi for topical application. aspergillus Allergen extracts were used to induce an allergic reaction in mice. A small portion of the mouse's back was shaved. A 1×1cm sterile gauze patch containing the allergen was attached to the skin using a bio-occlusive transparent dressing and a band-aid. The patch was left on the skin for a 7-day sensitization period until it fell off or was removed on the 8th day. This process was repeated twice, with a 2-week rest period in between.

[0179] Several days after the final sensitization, mice were exposed to 100 µL of allergen by applying one drop of allergen to each nostril.

[0180] HMO (more specifically, a 2:1 mixture of 2FL and LNnt) was incorporated into the low-fiber diet of the HMO group up to 5%. The volume and calories of the control diets of the other groups were adjusted with maltodextrin and cellulose.

[0181] A diet was started 3 weeks before the first sensitization and continued throughout the experiment. Feces were collected 3 weeks after feeding and before the first sensitization. At harvest, cecum and blood were collected and immunoglobulins were measured by ELISA as described previously by Holvoet et al. (Holvoet S. Allergy 2016 Dec;71(12):1753-1761): Specific IgE levels were quantified as follows: Aspergillus allergen (Greer Laboratories) was resuspended in distilled water containing antiprotease (Sigma) and Triton X100 (Sigma). The solution was sonicated on ice for 5 minutes and centrifuged at 1000 rpm for 10 minutes. The supernatant was collected and the total protein content was measured using the BCA Protein Assay Kit (ThermoScientific; Zug, Switzerland) according to the manufacturer's protocol. Following the manufacturer's protocol, total Aspergillus protein was labeled using digoxigenin-3-O-methylcarbonyl-e-aminohexanoic acid-N-hydroxysuccinimide ester (DIG; Roche, Basel, Switzerland). 96-well plates were coated with 100 µL of carbonate buffer containing 2 µg / mL rat anti-mouse IgE (BD-Bioscience) and incubated overnight at 4°C. The plates were washed with 0.05% Tween PBS and blocked for 1 h at room temperature with 200 µL PBS containing 10% fetal bovine serum (FCS; Bioconcept). The diluted serum was incubated for 2 h at room temperature. After washing, the DIG-labeled antigen was incubated for 90 min at room temperature. The plates were washed and incubated for 1 h at room temperature with HRP-labeled anti-DIG antibody (Roche, Switzerland). The reaction was performed using TMB and stopped with 1N HCl (Merck). The plate was read at 450 nm, and the results are expressed as OD.

[0182] Specific IgE levels measured in mice from different experimental groups were reported. Figure 3 middle.

[0183] Colonic propionate levels were measured as follows. Propionate in the cecum and feces of HMO-fed mice was analyzed by gas chromatography-mass spectrometry (GC-MS) using an Agilent Technologies 6890 Series XL MSD 5975 C (Santa Clara, CA) after ionization with an electron bombardment source. Briefly, the cecum and feces were first homogenized in a solution of orthophosphate and mercuric acetate D3 corresponding to 4 times the fecal weight. The sample was then homogenized with glass beads using a multivortex for 20 minutes. The preparation was centrifuged at 2,000 g for 15 minutes at 4 °C. The supernatant was collected and weighed. Ten μL of 37% HCl and 3 mL of chloroform were added, and homogenization was performed for 20 minutes. After centrifugation at 1,800 g for 10 minutes at 4 °C, the supernatant was removed. Add 10 µL of tert-butyldimethylsilylimidazolium and heat the sample at 60 °C for 30 minutes, then cool and inject the sample again. Freeze-dry to determine the moisture content, and the moisture content is used to calculate the SCFA per gram of dry material.

[0184] Specific SCFA levels measured in mice from different experimental groups were reported. Figure 4 and Figure 5 middle.

[0185] result: Figure 4 and Figure 5 The study showed a significant increase in propionate levels in mice supplemented with HMO. Based on IgE measurements, specific IgE levels were significantly lower in these mice than in the positive controls that did not receive HMO supplementation, highlighting the association between increased colonic propionate and reduced allergic sensitization.

Claims

1. A nutritional composition comprising at least one fucoidylated oligosaccharide and at least one N-acetylated oligosaccharide for preventing and / or treating allergic symptoms in an infant or young child by increasing propionate production in the infant or young child.

2. The nutritional composition according to claim 1, wherein the nutritional composition is used to prevent and / or treat allergic symptoms in an infant or young child by increasing the production of colonic propionate in the infant or young child.

3. A nutritional composition for use according to any one of the preceding claims, wherein the fucosylated oligosaccharide is selected from the list consisting of: 2'-fucosylvose, 3'-fucosylvose, difucosylvose, lactose-N-fucopentose I, lactose-N-fucopentose II, lactose-N-fucopentose III, lactose-N-fucopentose V, lactose-N-fucohexose, lactose-N-difucohexose I, fucosylvose-N-hexose, fucosylvose-N-neohexose I, fucosylvose-N-neohexose II, difucosylvose-N-hexose I, difucosylvose-N-neohexose I, difucosylvose-N-neohexose II, fucosylvose-P-lactose-N-hexose, and any combination thereof.

4. A nutritional composition for use according to any one of the preceding claims, wherein the fucoidylated oligosaccharide comprises a 2' fucoidyl epitope.

5. A nutritional composition for use according to any one of the preceding claims, wherein the fucoidylated oligosaccharide is 2'-fucosylated lactose (2'FL).

6. A nutritional composition for use according to any one of the preceding claims, wherein the N-acetylated oligosaccharide is selected from the list of the following: lactose-N-tetrasaccharide (LNT), lactose-N-neotetrasaccharide (LNnT), and any combination thereof.

7. A nutritional composition for use according to any one of the preceding claims, wherein the N-acetylated oligosaccharide is lactose-N-neotetrasaccharide (LNnT), p-lactose-N-neohexose (p-LNnH), or any combination thereof, preferably wherein the N-acetylated oligosaccharide is lactose-N-neotetrasaccharide (LNnT).

8. A nutritional composition for use according to any one of the preceding claims, comprising 2'-fucosyllactose (2'FL) and lactose-N-neotetrasaccharide (LNnT), or comprising a mixture of oligosaccharides consisting of 2'-fucosyllactose (2'FL) and lactose-N-neotetrasaccharide (LNnT).

9. A nutritional composition for use according to any one of the preceding claims, wherein the weight ratio of the fucoidylated oligosaccharide to the N-acetylated oligosaccharide is 1:10 to 12:1, such as 1:2 to 2:

1.

10. A nutritional composition for use according to any one of the preceding claims, wherein the at least one fucoidylated oligosaccharide and the at least one N-acetylated oligosaccharide are present in an amount of 0.1% to 10% by weight, such as 0.5% to 7% by weight or 1% to 5% by weight, of the nutritional composition.

11. A nutritional composition for use according to any one of the preceding claims, comprising at least one other oligosaccharide and / or fiber and / or human milk oligosaccharide precursor, selected from the list including: GOS, FOS, XOS, inulin, polydextrose, sialylated oligosaccharides, N-acetylated oligosaccharides, sialic acid, fucose, and any combination thereof.

12. A nutritional composition for use according to any one of the preceding claims, wherein it does not contain any sialylated oligosaccharides.

13. A nutritional composition for use according to any one of the preceding claims, said composition further comprising at least one probiotic, said probiotic being present in an amount of 10... 3 cfu / g of the composition up to 10 12 The composition described in cfu / g (dry weight) 14. A nutritional composition for use according to any one of the preceding claims, wherein the nutritional composition is infant formula, stage 1 infant formula, stage 2 infant formula or follow-up formula, baby food, infant cereal composition, fortifier or supplement.

15. A nutritional composition for use according to any one of the preceding claims, wherein the infant or young child is at risk of developing an allergic reaction, wherein the allergic reaction is selected from the group consisting of: allergic sensitization, food allergy, atopic dermatitis or eczema, asthma, wheezing, allergic rhinitis, rhinoconjunctivitis, eosinophilic esophagitis, hypersensitivity, allergy, urticaria, angioedema, food intolerance; and allergic reactions derived from allergenic proteins in food, pollen, animal dander, house dust mites, or venom.

16. A nutritional composition for use according to any one of the preceding claims, wherein the nutritional composition is a synthetic nutritional composition.

Images