Nutritional composition and use of nutritional composition

Abstract

ABSTRACT NON-THERAPEUTIC METHOD; USE OF A PROTEIN; NUTRITIONAL COMPOSITION; AND USE OF THE NUTRITIONAL COMPOSITION The present invention relates to fermented infant formulas comprising non-digestible oligosaccharides to improve intestinal tract health by decreasing the digestive effort of protein, by decreasing the amount of endogenously formed proteases concomitant with increased protein digestion, and reduced protein fermentation.

Description

[001] The present invention relates to nutritional formulas for infants and children and, in particular, aims at reducing proteolytic activity in the intestinal tract, while maintaining or even improving effective protein digestion. HISTORY OF THE INVENTION

[002] The digestion of dietary proteins is achieved by the release of proteases from the stomach and pancreas. The release of proteases is normally tightly controlled, thus ensuring that neither too few nor too many proteolytic enzymes are secreted. This is important since a high release of proteases and high proteolytic activity have several disadvantages. A very high release of proteases will result in energy loss and a loss of essential amino acids due to the fact that the proteases themselves are highly resistant to proteolytic activity and will enter the colon mostly unchanged. Subsequently, upon the entry of an increased amount of protein (in the form of proteases) into the colon, the intestinal microbiota will ferment the proteins resulting in a disadvantageously higher pH, a change in the composition of the intestinal microbiota, and the formation of toxic metabolites such as phenol, indole, and amines.This change in colon physiology can also lead to elevated intestinal infections. Finally, proteases, such as trypsin, are known to cleave receptor-activated proteins (PARs), such as PARII, and thus disrupt the integrity of the intestinal barrier. This can result in an increased perception of abdominal pain. Inflammatory diseases such as IBS-D and UC have been linked to this. Petition 870260046869, dated 05 / 18 / 2026, page 12 / 87 2 / 57 Elevated levels of proteolytic activity within the intestinal lumen and subsequent activation of PAR-II. Furthermore, increased fecal proteolytic activity is linked to diaper rashes. In general, a very high release of proteases will result in decreased gastrointestinal comfort, functional digestive disorders, gastrointestinal gas formation, and / or flatulence.

[003] Very low secretion of proteolytic activity, on the one hand, is disadvantageous since dietary proteins are not properly digested, leading to a loss of essential amino acids and energy, and an increased protein load in the colon.

[004] Especially in infants and children, strong regulation of protein digestion and release of proteolytic activity is of paramount importance. Firstly, for infants and children, limited protein loss is essential for proper growth and development. Loss of essential amino acids and energy impairs growth and development. Secondly, the intestinal barrier function in infants and children is still immature and the gut microbiota is still developing and therefore more susceptible to the disadvantages mentioned above. The perception of pain, for example due to colic or cramps, is often a major cause of concern and initiates many medical visits, and thus reducing the risk of perceived pain, for example, prevention and / or treatment of colic and / or cramps, is desirable.

[005] Known ways to improve protein digestion, particularly in infants, involve partial predigestion of dietary proteins by proteases. In addition Petition 870260046869, dated 05 / 18 / 2026, page 13 / 87 3 / 57 of this, Alm, 1982, J Dairy Sci 65:509-514 reports that a low pH of dairy products, especially as a result of fermentation, has a positive influence on the in vitro digestibility of proteins. It is considered that in many digestive disorders the secretion of hydrochloric acid is impaired and thus the suitability of such low pH dairy products is suggested for infants, children and adults. Vass et al, 1984, Acta Medica Hungarica, 41, 15-161 report that fermented milks have the highest protein utilization rate, defined as the increase in body mass in g per gram of protein intake, in weaned rats, and this is attributed to better protein digestibility.

[006] Gallia Lactofidus® is an acidified infant formula, resulting from fermentation by two specific strains of lactic acid bacteria. This formula is shown to facilitate protein digestibility and improve intestinal transit.

[007] Document WO 2009 / 151330 discloses fermented infant formula with non-digestible oligosaccharides for use in improved intestinal colonization in newborns delivered by cesarean section.

[008] US patent 2011 / 097437 discloses nutritional compositions comprising non-digestible oligosaccharides and a protein substrate fermented with Bifidobacterium breve and Streptococcus thermophilus for use in reducing bacterial translocation and improving intestinal barrier function.

[009] US patent 2010 / 278781 discloses a protein comprising a composition, fermented by Bifidobacterium breve and comprising non-oligosaccharides Petition 870260046869, dated 05 / 18 / 2026, page 14 / 87 4 / 57 digestible foods with various health benefits. SUMMARY OF THE INVENTION

[010] The inventors discovered that by employing guinea pigs as an animal model, through consumption of a fermented formula, the amount of endogenous proteolytic enzymes detected in the terminal ileum was significantly reduced compared to the amount detected through consumption of a standard non-fermented formula. Surprisingly, the amount of endogenous proteolytic enzymes was also significantly reduced compared to the amount detected through consumption of a formula with extensively hydrolyzed (i.e., pre-digested) proteins.

[011] Furthermore, and also unexpectedly, apparent and true protein digestibility was highest in guinea pigs that also consumed the fermented formula, despite their higher daily protein intake. This is indicative of reduced digestive effort. Digestive effort is defined as the amount of protease activity secreted per gram of protein ingested. Digestive efficiency is defined as the reciprocal value of digestive effort. This means that at high digestive efficiency (little protease needed to digest the ingested protein), digestive effort is low.

[012] The inventors also found that proteolytic activity in fecal samples from exclusively breastfed human infants was lower than in infants fed a standard formula. When the formula administered to human infants comprised non-digestible oligosaccharides, proteolytic activity in fecal samples was lower, with similar values ​​as observed for human infants. Petition 870260046869, dated 05 / 18 / 2026, page 15 / 87 5 / 57 breastfed.

[013] Thus, by feeding a protein-comprising fermented formula, the release of endogenous protease, digestive effort, loss of endogenous protein in the small intestine, and the protein load entering the colon are advantageously reduced. The additional presence of non-digestible oligosaccharides reduces the proteolytic activity of the microbiota in the colon. Therefore, a formula for infants and children that is fermented and comprises non-digestible oligosaccharides will have an additional enhanced effect in reducing protein fermentation and proteolytic activity in the colon. Furthermore, a formula for infants or children that is fermented and comprises non-digestible oligosaccharides will have a beneficial effect in reducing proteolytic activity throughout the entire intestinal tract, i.e., large and small intestines.Therefore, a fermented formula comprising non-digestible oligosaccharides is advantageously used as nutrition for infants or children for use in the prevention and / or treatment of diaper rash or in promoting tract health by reducing digestive effort, improving the effectiveness of protein digestion, reducing endogenous protein loss, reducing endogenous protease secretion, reducing colon protein fermentation, and / or reducing the protein load entering the colon.

[014] The discovery that the efficiency of protein digestion is increased and endogenous protein loss is also advantageously reduced allows for the formulation of an infant formula [or for children] with lower protein concentrations than those used until now. Petition 870260046869, dated 05 / 18 / 2026, page 16 / 87 6 / 57 DETAILED DESCRIPTION OF THE INVENTION

[015] The present invention relates to a method, in particular, a non-therapeutic method, for a) to increase the efficiency of protein digestion, b) reduce the secretion of proteases in the small and / or large intestine, c) reduce proteolytic activity in the small and / or large intestine, d) reduce the protein load entering the large intestine and / or (e) to reduce protein fermentation in the large intestine in a human subject aged 0 to 36 months, the method comprising administering a nutritional composition comprising - protein in an amount of 5 to 20% by weight based on the dry weight of the nutritional composition, and in an amount of 1.6 to 4.0 g per 100 kcal, - Non-digestible oligosaccharides in an amount of 0.5 to 20% by weight based on the dry weight of the nutritional composition. - at least 10% by weight based on the dry weight of the nutritional composition of a protein comprising a composition fermented by lactic acid bacteria, and - 0.10 to 1.5% by weight of the sum of lactate and lactic acid based on the dry weight of the nutritional composition, where the sum of L-lactic acid and L-lactate is more than 50% by weight based on the sum of total lactic acid and lactate, for a human individual aged 0 to 36 months.

[016] In a preferred embodiment, the present Petition 870260046869, dated 05 / 18 / 2026, p. 17 / 87 7 / 57 method is for f) reducing digestive effort.

[017] Preferably, the increase or decrease of a, b, c, de / or ee / or f) is compared to the effect of a nutritional composition not comprising a protein comprising a composition fermented by lactic acid bacteria and non-digestible oligosaccharides.

[018] The invention also relates to a method for promoting intestinal tract health in a human individual aged 0 to 36 months, comprising administering to the infant a nutritional composition comprising - protein in an amount of 5 to 20% by weight based on the dry weight of the nutritional composition, and in an amount of 1.6 to 4.0 g per 100 kcal, - Non-digestible oligosaccharides in an amount of 0.5 to 20% by weight based on the dry weight of the nutritional composition. - at least 10% by weight based on the dry weight of the nutritional composition of a protein comprising a composition fermented by lactic acid bacteria, and - 0.10 to 1.5% by weight of the sum of lactate and lactic acid based on the dry weight of the nutritional composition, where the sum of L-lactic acid and L-lactate is more than 50% by weight based on the sum of total lactic acid and lactate.

[019] In other words, the invention relates to the use of a protein comprising a composition fermented by lactic acid bacteria and non-digestible oligosaccharides in the manufacture of a nutritional composition to promote the health of the intestinal tract in a human individual aged 0 to 36 months, wherein the nutritional composition comprises - protein in an amount of 5 to 20% by weight with Petition 870260046869, dated 05 / 18 / 2026, page 18 / 87 8 / 57 based on the dry weight of the nutritional composition, and in an amount of 1.6 to 4.0 g per 100 kcal, - Non-digestible oligosaccharides in an amount of 0.5 to 20% by weight based on the dry weight of the nutritional composition. - at least 10% by weight based on the dry weight of the nutritional composition of a protein comprising a composition fermented by lactic acid bacteria, and - 0.10 to 1.5% by weight of the sum of lactate and lactic acid based on the dry weight of the nutritional composition, where the sum of L-lactic acid and L-lactate is more than 50% by weight based on the sum of total lactic acid and lactate.

[020] The invention can also be formulated as a nutritional composition comprising - protein in an amount of 5 to 20% by weight based on the dry weight of the nutritional composition, and in an amount of 1.6 to 4.0 g per 100 kcal, - Non-digestible oligosaccharides in an amount of 0.5 to 20% by weight based on the dry weight of the nutritional composition. - at least 10% by weight based on the dry weight of the nutritional composition of a protein comprising a composition fermented by lactic acid bacteria, and - 0.10 to 1.5% by weight of the sum of lactate and lactic acid based on the dry weight of the nutritional composition, where the sum of L-lactic acid and L-lactate is more than 50% by weight based on the sum of total lactic acid and lactate,

[021] To promote intestinal tract health in a human individual aged 0 to 36 months.

[022] Preferably, promote the health of Petition 870260046869, dated 05 / 18 / 2026, page 19 / 87 9 / 57 intestinal tract is through a) increased efficiency of protein digestion, b) reduction in protease secretion in the small and / or large intestine, c) reduction of proteolytic activity in the small and / or large intestine, d) reduction of the protein load entering the large intestine and / or e) reduction of protein fermentation in the large intestine. where the increase or decrease is compared to the effect of a nutritional composition not comprising a protein comprising a composition fermented by lactic acid bacteria and non-digestible oligosaccharides.

[023] The invention also relates to a method of treating and / or preventing diaper rash or colic and / or cramps in a human individual aged 0 to 36 months, comprising administering to the baby a nutritional composition comprising - protein in an amount of 5 to 20% by weight based on the dry weight of the nutritional composition, and in an amount of 1.6 to 4.0 g per 100 kcal, - Non-digestible oligosaccharides in an amount of 0.5 to 20% by weight based on the dry weight of the nutritional composition. - at least 10% by weight based on the dry weight of the nutritional composition of a protein comprising a composition fermented by lactic acid bacteria, and - 0.10 to 1.5% by weight of the sum of lactate and lactic acid based on the dry weight of the nutritional composition and in Petition 870260046869, dated 05 / 18 / 2026, p. 20 / 87 10 / 57 that the sum of L-lactic acid and L-lactate is more than 50% by weight based on the sum of total lactic acid and lactate.

[024] In other words, the invention relates to the use of a protein comprising a composition fermented by lactic acid bacteria and non-digestible oligosaccharides in the manufacture of a nutritional composition for the treatment and / or prevention of diaper rash or colic and / or cramps in a human individual aged 0 to 36 months, wherein the nutritional composition comprises - protein in an amount of 5 to 20% by weight based on the dry weight of the nutritional composition, and in an amount of 1.6 to 4.0 g per 100 kcal, - Non-digestible oligosaccharides in an amount of 0.5 to 20% by weight based on the dry weight of the nutritional composition. - at least 10% by weight based on the dry weight of the nutritional composition of a protein comprising a composition fermented by lactic acid bacteria, and - 0.10 to 1.5% by weight of the sum of lactate and lactic acid based on the dry weight of the nutritional composition, where the sum of L-lactic acid and L-lactate is more than 50% by weight based on the sum of total lactic acid and lactate.

[025] The invention can also be formulated as a nutritional composition comprising - protein in an amount of 5 to 20% by weight based on the dry weight of the nutritional composition, and in an amount of 1.6 to 4.0 g per 100 kcal, - Non-digestible oligosaccharides in an amount of 0.5 to 20% by weight based on the dry weight of the nutritional composition. Petition 870260046869, dated 05 / 18 / 2026, page 21 / 87 11 / 57 - at least 10% by weight based on the dry weight of the nutritional composition of a protein comprising a composition fermented by lactic acid bacteria, and - 0.10 to 1.5% by weight of the sum of lactate and lactic acid based on the dry weight of the nutritional composition, wherein the sum of L-lactic acid and L-lactate is more than 50% by weight based on the sum of total lactic acid and lactate, for use in the treatment and / or prevention of diaper rash or colic and / or cramps in a human individual aged 0 to 36 months.

[026] The invention also relates to a nutritional composition comprising protein, lipids, digestible carbohydrates and non-digestible oligosaccharides, wherein the protein is present in an amount of 5 to 20% by weight based on the dry weight of the nutritional composition and in an amount of 1.6 to 4.0 g per 100 kcal, wherein the lipids are present in an amount of 3 to 7 g per 100 kcal, wherein the digestible carbohydrates are present in an amount of 5 to 20 g per 100 kcal, wherein the non-digestible oligosaccharides are present in an amount of 0.5 to 20% by weight based on the dry weight of the nutritional composition, further characterized in that the nutritional composition comprises at least 10% by weight based on the dry weight of the nutritional composition of a protein comprising a composition fermented by lactic acid bacteria, and characterized in that the nutritional composition comprises 0.10 to 1,5% by weight of the sum of lactate and lactic acid based on the dry weight of the nutritional composition, and where the sum of L-lactic acid and L-lactate is more than 50% by weight based on the sum of the total lactic acid and lactate. Petition 870260046869, dated 05 / 18 / 2026, p. 22 / 87 12 / 57

[027] The invention also relates to a method for providing nutrition to a human individual aged 0 to 36 months, the method comprising administering a nutritional composition according to the invention to the human individual.

[028] In other words, the invention relates to the use of a nutritional composition according to the present invention to provide nutrition to a human individual aged 0 to 36 months.

[029] The invention can also be formulated as a nutritional composition according to the present invention for use in providing nutrition to a human individual aged 0 to 36 months.

[030] Preferably according to the present invention, the nutritional composition has a viscosity when administered close to that of human milk. Thus, in an embodiment according to the present invention, the nutritional composition does not comprise a thickener, preferably it does not comprise a thickener selected from the group consisting of locust bean gum, tara gum, tragacanth gum, guar gum, and fenugreek gum, preferably it does not comprise any of locust bean gum, tara gum, tragacanth gum, guar gum or fenugreek gum.

[031] In one aspect, the invention relates to a method for reducing proteolytic activity in the intestine of a human subject aged 0 to 36 months, said method comprising administering a nutritional composition to said human subject comprising protein, lipids, digestible carbohydrates and non-digestible oligosaccharides, wherein the protein is present in an amount of 5 to 20% by weight based on the dry weight of the nutritional composition, and in Petition 870260046869, dated 05 / 18 / 2026, page 23 / 87 13 / 57 an amount of 1.6 to 4.0 g per 100 kcal, wherein lipids are present in an amount of 3 to 7 g per 100 kcal, wherein digestible carbohydrates are present in an amount of 5 to 20 g per 100 kcal, wherein non-digestible oligosaccharides are present in an amount of 0.5 to 20% by weight based on the dry weight of the nutritional composition, and wherein the non-digestible oligosaccharides are selected from the group consisting of transgalacto-oligosaccharides, fructooligosaccharides and galacturonic acid oligosaccharides, more preferably, the non-digestible oligosaccharides comprise galacturonic acid.

[032] For clarity, it is noted that the “nutritional composition mentioned above” refers to the final nutritional composition that should be ingested or administered, and the “protein comprising composition fermented by lactic acid bacteria” is included in the nutritional composition. The nutritional composition may thus also be referred to as “final nutritional composition” or “total nutritional composition.” The “protein comprising composition fermented by lactic acid bacteria” may also be referred to as “protein comprising fermented composition” or “fermented ingredient.” Also, for clarity, it is noted that the “protein” portion of the “protein comprising composition fermented by lactic acid bacteria” is included in the “5 to 20% by weight of the protein in the nutritional composition.”

[033] The term “nutritional composition” used throughout this report, that is, the final nutritional composition to be ingested or administered, also refers to the “nutritional composition for a human individual aged Petition 870260046869, dated 05 / 18 / 2026, p. 24 / 87 14 / 57 to 36 months as specified above. A human individual aged 0 to 36 months includes infants and children and may thus also be referred to as a baby or child. Also, note that wherever words such as “the present nutritional composition” or “nutritional composition according to the (present) invention” are used in the present description, this also refers to the methods and uses according to the present invention. FERMENTED INGREDIENT

[034] Fermentation is the process of deriving energy from the oxidation of carbohydrates, such as lactose found in milk, using an endogenous electron acceptor, which is normally an organic compound. This is in contrast to cellular respiration, where electrons are donated to an endogenous electron acceptor, such as oxygen, via an electron transport chain.

[035] In the present invention, the fermentation of a milk-derived product by lactic acid-producing bacteria has the common meaning of converting the carbohydrates present in the milk-derived product into organic acids. These organic acids formed may comprise, in addition to lactic acid, other organic acids such as acetate. The carbohydrate that is fermented is preferably lactose.

[036] Lactic acid bacteria are also referred to as lactic acid-producing bacteria and include bacteria of the genera Streptococcus, Lactococcus, Lactobacillus, Leuconostoc, Enterococcus, Oenococcus, Pediococcus, and Bifidobacterium.

[037] The present nutritional composition Petition 870260046869, dated 05 / 18 / 2026, p. 25 / 87 15 / 57 comprises a protein comprising a composition fermented by lactic acid bacteria. The present nutritional composition preferably comprises a protein derived from fermented milk comprising a composition. This protein derived from fermented milk comprising a composition is obtained by incubating a combination of milk, for example, skimmed milk, or a milk product, for example, whey, with at least one strain of lactic acid bacteria, such as lactococci, lactobacilli, streptococci and bifidobacteria, preferably the protein derived from fermented milk comprising a composition is obtained by incubation with at least one selected strain of lactococci, lactobacilli and streptococci, preferably with at least one selected strain of streptococci.Preferably, lactic acid bacteria that perform homolactic fermentation are used for fermentation, since in this case two lactic acids are produced per unit of sugar and no gas is formed. Homolactic lactic acid bacteria include Streptococcus thermophilus, Lactococcus species, preferably Lactococcus lactis, and Group I lactobacilli such as Lactobacillus acidopilus, Lactobacillus helveticus and Lactobacillus salivarius, and faculatively heterofermentative lactobacilli that produce two lactic acids from hexose sugars, including Lactobacillus casei, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus plantarum, and Lactobacillus sakei. Thus, in one embodiment, preferably the fermented milk-derived protein comprising the composition is obtained by incubating a combination of milk, for example, skim milk, or a milk-derived product, for example, whey, with... Petition 870260046869, dated 05 / 18 / 2026, page 26 / 87 16 / 57 at least one selected strain of Streptococcus thermophilus, Lactococcus lactis, Lactobacillus acidopilus, Lactobacillus helveticus, Lacobacillus salivarius, Lacobacillus casei, Lacobacillus paracasei, Lacobacillus rhamnosus, Lacobacillus plantarum and Lacobacillus sakei, preferably the protein derived from fermented milk comprising the composition is obtained by incubation with Streptococcus thermophilus. Preferably, the mixture is incubated for 10 minutes to about 6 hours. The temperature during incubation is preferably between 20 and 50 °C. In one embodiment, after incubation, the incubated composition is preferably subjected to a heat treatment. By this heat treatment, preferably at least 90% of live microorganisms are inactivated, more preferably at least 95%. Preferably, the fermented nutritional composition comprises less than 110⁵ colony-forming units (cfu) of live lactic acid bacteria / g of dry weight. Heat treatment is preferably carried out at a temperature between 80 and 180 °C. Inactivation of the lactic acid bacteria advantageously results in less post-acidification and a safer product. This is especially advantageous when the nutritional composition is to be administered to infants and children. Procedures for preparing fermented ingredients suitable for the purpose of the present invention are known per se. Document EP 778885, which is incorporated herein by reference, discloses, in particular in Example 7, a suitable process for preparing a fermented composition. Document FR 2723960, which is incorporated herein by reference, discloses, in particular in Example 6, a suitable process for preparing a fermented composition. Petition 870260046869, dated 05 / 18 / 2026, page 27 / 87 17 / 57

[038] Briefly, a milk-derived product, preferably pasteurized, containing lactose and optionally other macronutrients such as fats, preferably vegetable fats, casein, whey protein, vitamins and / or minerals, etc., is concentrated, for example, between 15 and 50% dry matter and then inoculated with S. thermophilus, for example, with 5% of a culture containing 10⁶ to 10¹⁰ bacteria per ml. Preferably, this milk-derived product comprises milk protein peptides. The temperature and duration of fermentation are as mentioned above. Appropriately after fermentation, the fermented protein comprising the composition may be pasteurized or sterilized and, for example, spray-dried or freeze-dried to provide a form suitable for formulation into the final product.

[039] The bacterial strains of S. thermophilus that are preferably used to prepare the fermented protein comprising the composition for the purpose of the present invention develop beta-galactosidase activity during the fermentation of the substrate. Preferably, the beta-galactosidase activity develops in parallel with acidity. The selection of a suitable strain of S. thermophilus is described in Example 2 of EP 778885 and Example 1 of FR 2723960. It is preferred that in the nutritional composition according to the invention and in its use, at least part of the beta-galactosidase activity that developed during fermentation is retained. Preferably, at least part of the beta-galactosidase activity that is retained is lactase activity. Upon digestion in the human individual, the lactase activity in the nutritional composition has a beneficial effect on promoting Petition 870260046869, dated 05 / 18 / 2026, p. 28 / 87 18 / 57 of intestinal tract health. Preferably according to the present invention, per gram of dry weight, the nutritional composition comprises 0.3 to 4 U of beta-galactosidase activity, preferably the nutritional composition comprises 0.3 to 4 U per gram of dry weight of lactase activity. In a further preferred embodiment according to the present invention, the nutritional composition comprises 10²-10⁵ cfu of live S. thermophilus bacteria, preferably the nutritional composition comprises 10²-10⁴ of live S. thermophilus bacteria.

[040] The preferred strains of S. thermophilus for preparing protein comprising fermented composition, preferably protein comprising composition derived from fermented milk for the purpose of the present invention were deposited by Compagnie Gervais Danone in the Collection Nationale de Cultures de Microorganismes (CNCM) run by the Institut Pasteur, 25 rue du Docteur Roux, Paris, France on August 23, 1995 under accession number I-1620 and on August 25, 1994 under accession number I-1470.

[041] Preferably, in the preparation of the protein comprising fermented composition, other strains of lactic acid bacteria are additionally present, or simultaneously or consecutively, the composition is additionally fermented by other strains of lactic acid bacteria. Other strains of lactic acid bacteria are preferably selected from the group consisting of Lactobacillus and Bifidobacteria, more preferably Bifidobacterium breve, more preferably the Bifidobacterium breve strain deposited by Compagnie Gervais Danone in the CNCM under number I-2219 on May 31, 1999. Petition 870260046869, dated 05 / 18 / 2026, p. 29 / 87 19 / 57

[042] In one embodiment, the protein comprising composition fermented by lactic acid bacteria, comprised Streptococcus thermophilus, and / or Bifidobacterium breve.

[043] In one embodiment, the protein comprising composition fermented by lactic acid bacteria is fermented by Streptococcus thermophilus, and / or Bifidobacterium breve.

[044] In one embodiment, the nutritional composition comprises protein comprising composition fermented by lactic acid bacteria wherein the lactic acid bacteria are inactivated after fermentation.

[045] Preferably, the present protein comprising fermented composition is not fermented by Lactobacillus bulgaricus. Fermented products of L. bulgaricus are considered unsuitable for infants, since in young infants, the specific dehydrogenase that converts D-lactate to pyruvate is much less active than the dehydrogenase that converts L-lactate.

[046] The fermented composition comprising protein comprises protein. The protein is preferably selected from the group consisting of non-human animal proteins, preferably milk proteins. The fermented composition comprising protein preferably contains casein and / or whey protein, more preferably bovine whey proteins and / or bovine casein. The fermented composition comprising protein preferably comprises casein and whey proteins in a casein:whey protein weight ratio of 10:90 to 90:10, more preferably 20:80 to 80:20, even more Petition 870260046869, dated 05 / 18 / 2026, page 30 / 87 20 / 57 preferably 35:65 to 55:45.

[047] The fermented composition comprising protein preferably comprises 1.25 to 4 g of protein per 100 kcal of the fermented composition comprising protein, preferably 1.5 to 3 g, even more preferably 1.7 to 2.5 g per 100 kcal. When in liquid form, the fermented composition comprising protein preferably comprises 0.5 to 6.0 g, more preferably 1.0 to 3.0 g, even more preferably 1.0 to 1.5 g of protein per 100 ml, most preferably 1.0 to 1.3 g of protein per 100 ml. Based on dry weight, the present fermented composition comprising protein preferably comprises 5 to 20% by weight of protein, preferably at least 8% by weight, more preferably 8 to 14% by weight, the protein even more preferably 8 to 9.5% by weight based on the dry weight of the fermented composition comprising protein.

[048] Protein and carbohydrates are considered to have a caloric density of 4 kcal / g, fat of 9 kcal / g and non-digestible oligosaccharides of 2 kcal / g.

[049] The present nutritional composition preferably comprises 10 to 100% by weight of the fermented composition comprising protein by lactic acid bacteria, preferably a composition derived from fermented milk comprising protein, based on the dry weight of the total nutritional composition. In one embodiment, the present nutritional composition preferably contains 15 to 70% by weight, preferably 15 to 50% by weight, based on the dry weight of the final nutritional composition of the fermented composition comprising protein by lactic acid bacteria. Petition 870260046869, dated 05 / 18 / 2026, page 31 / 87 21 / 57 Higher concentrations of the fermented composition comprising protein advantageously improve the efficiency of protein digestion.

[050] The pH of the present nutritional composition is preferably between 5.0 and 7.5, more preferably between 5.0 and 7.0, even more preferably between 5.0 and 6.0, most preferably between 5.5 and 6.0. Preferably, the present nutritional composition is a liquid having a pH of 5.5 to 6.0. The present nutritional composition comprises lactic acid and / or lactate. Lactic acid and / or lactate is formed by fermentation by lactic acid bacteria. Preferably, the present nutritional composition comprises between 0.1 and 1.5% by weight of lactic acid and / or lactate, more preferably between 0.2 and 1.0% by weight, based on the dry weight of the nutritional composition. The more lactate is present, the more the nutritional composition comprises the fermented composition comprising protein. Preferably at least 50% by weight, even more preferably at least 90% by weight, of the sum of lactic acid and lactate is in the form of the L-isomer.Thus, in one embodiment, the sum of L-lactic acid and L-lactate is more than 50% by weight, more preferably more than 90% by weight, based on the sum of total lactic acid and lactate. L-lactate and L-lactic acid are the same as L-(+)-lactate and L(+)lactic acid.

[051] In one embodiment, the nutritional composition for infants and children comprises 0.10 to 1.5% by weight of the sum of lactate and lactic acid based on the dry weight of the nutritional composition, wherein the sum of L-lactic acid and L-lactate is more than 50% by weight based on the sum of the total Petition 870260046869, dated 05 / 18 / 2026, p. 32 / 87 22 / 57 lactic acid and lactate. PROTEIN COMPONENT

[052] The present nutritional composition comprises a protein component. The protein used in the nutritional composition is preferably selected from the group consisting of non-human animal proteins, preferably milk proteins, vegetable proteins, such as preferably soy protein and / or rice protein, and mixtures thereof. The present nutritional composition preferably contains casein and / or whey protein, more preferably bovine whey protein and / or bovine casein. Thus, in one embodiment, the protein component comprises protein selected from the group consisting of whey protein and casein, preferably whey protein and casein, preferably the whey protein and / or casein is from cow's milk. Preferably, the protein comprises less than 5% by weight on a total protein basis of free amino acids, dipeptides, tripeptides or hydrolyzed protein.The present nutritional composition preferably comprises casein and whey proteins in a casein:whey protein weight ratio of 10:90 to 90:10, more preferably 20:80 to 80:20, and even more preferably 35:65 to 55:45.

[053] The % by weight of protein based on the dry weight of the present nutritional composition is calculated according to a Kjeldahl method by measuring the total nitrogen and using a conversion factor of 6.38 in the case of casein, or a conversion factor of 6.25 for proteins other than casein. The term 'protein' or 'protein component' as used in the present invention refers to the sum of Petition 870260046869, dated 05 / 18 / 2026, page 33 / 87 23 / 57 proteins, peptides and free amino acids.

[054] The present nutritional composition comprises protein providing 1.6 to 4.0 g of protein per 100 kcal of the nutritional composition, preferably providing 1.6 to 3.5 g, even more preferably 1.75 to 2.5 g per 100 kcal of the nutritional composition. In one embodiment, the present nutritional composition comprises protein providing 1.6 to 2.1 g of protein per 100 kcal of the nutritional composition, preferably providing 1.6 to 2.0 g, more preferably 1.75 to 2.1 g, even more preferably 1.75 to 2.0 g per 100 kcal of the nutritional composition. In one embodiment, the nutritional composition for infants and children according to the invention comprises protein preferably in an amount of less than 2.0 g per 100 kcal, preferably providing 1.6 to 1.9 g, even more preferably 1.75 to 1.85 g per 100 kcal of the nutritional composition.A very low protein content based on total calories will result in less adequate growth and development in infants and young children. When in liquid form, for example, as a ready-to-feed liquid, the nutritional composition preferably comprises 0.5 to 6.0 g, more preferably 1.0 to 3.0 g, even more preferably 1.0 to 1.5 g of protein per 100 ml, most preferably 1.0 to 1.3 g of protein per 100 ml. Based on dry weight, the present nutritional composition comprises 5 to 20% protein by weight, preferably at least 8% protein by weight based on the dry weight of the total nutritional composition, more preferably 8 to 14% by weight, even more preferably 8 to 9.5% protein by weight based on the dry weight of the total nutritional composition. Petition 870260046869, dated 05 / 18 / 2026, page 34 / 87 24 / 57

[055] Since the use of the present nutritional composition results in increased efficiency of protein digestion and reduced digestive effort of protein, the amount of protein based on total calories, based on 100 ml or based on the dry weight of the composition can advantageously be lower than that of standard infant formula. Currently, the protein concentration in infant formulas is higher than in human milk in order to ensure adequate absorption of essential amino acids. An increased protein concentration, however, has disadvantages as mentioned in the “application” section; it imposes a very heavy load on the kidneys of infants and is also correlated with obesity and other adverse health effects later in life. The term 'protein' or 'protein component' as used in the present invention refers to the sum of proteins, peptides and free amino acids. Non-digestible oligosaccharides

[056] The present nutritional composition comprises non-digestible oligosaccharides. Non-digestible oligosaccharides have been found to reduce proteolytic activity in the colon. Advantageously and more preferably, the non-digestible oligosaccharides are water-soluble (according to a method disclosed in L. Prosky et al, J. Assoc. Anal. Chem 71: 1017-1023, 1988) and are preferably oligosaccharides with a degree of polymerization (DP) of 2 to 200. The average DP of the non-digestible oligosaccharides is preferably below 200, more preferably below 100, even more preferably below 60, most preferably below 40. The non-digestible oligosaccharides are not digested in the intestine by the action of enzymes. Petition 870260046869, dated 05 / 18 / 2026, page 35 / 87 25 / 57 Digestive substances present in the human upper digestive tract (small intestine and stomach). Non-digestible oligosaccharides are fermented by the human intestinal microbiota. For example, glucose, fructose, galactose, sucrose, lactose, maltose, and maltodextrins are considered digestible. The raw materials of the oligosaccharide may comprise monosaccharides such as glucose, fructose, fucose, galactose, rhamnose, xylose, glucuronic acid, GalNac, etc., but these are not part of the oligosaccharides as in the present invention.

[057] The non-digestible oligosaccharides included in the nutritional compositions and methods according to the present invention preferably include a mixture of non-digestible oligosaccharides. It has been found that mixtures of non-digestible oligosaccharides reduce proteolytic activity in the colon.

[058] Non-digestible oligosaccharides are preferably selected from the group consisting of fructooligosaccharides, such as inulin, non-digestible dextrins, galacto-oligosaccharides, such as transgalactooligosaccharides, xylo-oligosaccharides, arabinooligosaccharides, arabinogalacto-oligosaccharides, glucooligosaccharides, gentio-oligosaccharides, glucomannooligosaccharides, galactomannanoligosaccharides, mannanoligosaccharides, isomalto-oligosaccharides, nigerooligosaccharides, glucomannanoligosaccharides, chitooligosaccharides, soy oligosaccharides, uronic acid oligosaccharides, sialyloligosaccharides, such as 3-sialyllactose (3-SL), 6-SL, lactosialyltesaccharide (LST) a,b,c, disialyllactoNtetraose (DSLNT), sialyl-lactoNhexaose (SLNH), DS-LNH, and fuco-oligosaccharides, such as Petition 870260046869, dated 05 / 18 / 2026, page 36 / 87 26 / 57 fucoidan (non)sulfated oligosaccharides, 2'-fucosyllactose (2'-FL), 3-FL, difucosyllactose, lacto-N-fucopenase, (LNFP) I, II, III, V, Lacto-N-neofucopenaose (LNnFP), Lacto-N-difucosyl-hexaose (LNDH), and mixtures thereof, even more preferably selected from the group consisting of fructo-oligosaccharide, such as inulin, galacto-oligosaccharide, such as transgalactooligosaccharide, uronic acid oligosaccharide and fucooligosaccharide and mixtures thereof, even more preferably transgalacto-oligosaccharide, inulin and / or uronic acid oligosaccharide, more preferably transgalacto-oligosaccharides. In an embodiment of the composition or methods according to the present invention, the non-digestible oligosaccharides are selected from the group consisting of transgalacto-oligosaccharides, fructooligosaccharides and galacturonic acid oligosaccharides and mixtures thereof.

[059] Non-digestible oligosaccharides are preferably selected from the group consisting of β-galacto-oligosaccharide, α-galacto-oligosaccharide, and galactan. According to a more preferred embodiment, the non-digestible oligosaccharides are β-galactooligosaccharide. Preferably, the non-digestible oligosaccharides comprise galacto-oligosaccharides with β(1,4), β(1,3) and / or β(1,6) glycosidic linkages and a terminal glucose. Transgalacto-oligosaccharide is, for example, available under the trade names Vivinal®GOS (Borculo Domo Ingredients, Zwolle, Netherlands), Bi2muno (Clasado), Cupoligo (Nissin Sugar) and Oligomate55 (Yakult).

[060] Non-digestible oligosaccharides Petition 870260046869, dated 05 / 18 / 2026, page 37 / 87 27 / 57 preferably comprise fructo-oligosaccharides. A fructo-oligosaccharide may, in another context, have names such as fructopolysaccharides, oligofructose, polyfructose, polyfructan, inulin, levan, and fructan, and may refer to oligosaccharides comprising β-linked fructose units, which are preferably linked by β(2,1) and / or β(2,6) glycosidic linkages, and a preferably DP between 2 and 200. Preferably, the fructo-oligosaccharide contains a glucose linked by a terminal β(2,1) glycosidic linkage. Preferably, the fructo-oligosaccharide contains at least 7 β-linked fructose units. In a further preferred embodiment, inulin is used. Inulin is a type of fructooligosaccharide in which at least 75% of the glycosidic linkages are β(2,1) linkages. Typically, inulin has an average chain length between 8 and 60 monosaccharide units.A suitable fructooligosaccharide for use in the compositions of the present invention is commercially available under the trade name Raftiline®HP (Orafti). Other suitable sources are raftilose (Orafti), fibrulose and fibrulin (Cosucra), and Frutafit and frutalose (Sensus).

[061] Preferably, the mixture of galactooligosaccharides and fructo-oligosaccharides is present in a weight ratio of 1 / 99 to 99 / 1, more preferably 1 / 19 to 19 / 1, even more preferably 1 to 19 / 1. This weight ratio is particularly advantageous when the non-digestible oligosaccharide A has a low DP and the non-digestible oligosaccharide B has a relatively high DP.

[062] Preferably, the mixture of short-chain fructooligosaccharides and long-chain fructooligosaccharides is present in a weight ratio of 1 / 99 to 99 / 1, Petition 870260046869, dated 05 / 18 / 2026, page 38 / 87 28 / 57 more preferably from 1 / 19 to 19 / 1, even more preferably from 1 to 19 / 1.

[063] In a more preferred embodiment, the present nutritional composition further comprises uronic acid oligosaccharides. The term uronic acid oligosaccharide as used in the present invention refers to an oligosaccharide in which at least 50% of the number of monosaccharide units present in the oligosaccharide is selected from the group consisting of guluronic acid, mannuronic acid, galacturonic acid, iduronic acid, riburonic acid, and glucuronic acid. In a preferred embodiment, the uronic acid oligosaccharide comprises at least 50% of the number of galacturonic acid based on the total uronic acid units in the uronic acid oligosaccharide.The uronic acid oligosaccharides used in the invention are preferably prepared from the degradation of pectin, pectate, alginate, chondroitin, hyaluronic acid, heparin, heparan, bacterial carbohydrates, and / or sialoglycans, more preferably from pectin and / or alginate, even more preferably from pectin, and even more preferably from polygalacturonic acid. Preferably, the degraded pectin is prepared by hydrolysis and / or beta-elimination of fruit and / or vegetable pectins, more preferably apple, citrus, and / or beet pectin, even more preferably apple, citrus, and / or beet pectin degraded by at least one lyase. Preferably, the non-digestible oligosaccharide is galacturonic acid oligosaccharide.

[064] In a preferred embodiment, at least one of the uronic acid terminal units of the oligosaccharide Petition 870260046869, dated 05 / 18 / 2026, page 39 / 87 29 / 57 uronic acid has a double bond. The double bond effectively protects against the attachment of pathogenic bacteria to intestinal epithelial cells. This is advantageous for infants and children. Preferably, one of the terminal units of uronic acid comprises the C4-C5 double bond. The double bond in the terminal unit of uronic acid can, for example, be obtained by enzymatic hydrolysis of pectin with lyase. Uronic acid oligosaccharide can be derivatized. Uronic acid oligosaccharide can be methoxylated and / or amidated.

[065] Preferably, the present nutritional composition comprises the non-digestible oligosaccharides transgalacto-oligosaccharide, fructo-oligosaccharide and a pectin degradation product.

[066] The weight ratio of transgalactooligosaccharide:fructo-oligosaccharide:pectin degradation product is preferentially (20 to 2):1:(1 to 3), more preferably (12 to 7) : 1 : (1 to 2).

[067] Preferably, the present invention relates to a nutritional composition, wherein the non-digestible oligosaccharide is selected from the group consisting of fructo-oligosaccharide, galacto-oligosaccharide and uronic acid oligosaccharide, wherein the uronic acid oligosaccharide is preferably galacturonic acid oligosaccharide.

[068] Preferred is a mixture of transgalactooligosaccharide with an average DP below 10, preferably below 6, and a fructo-oligosaccharide with an average DP below 10, preferably below 6. More preferred is a mixture of fructo-oligosaccharide with an average DP below 10, Petition 870260046869, dated 05 / 18 / 2026, page 40 / 87 30 / 57 preferably below 6 and a fructooligosaccharide with an average DP above 7, preferably above 11, even more preferably above 20. Such a mixture synergistically reduces protein fermentation in the colon and proteolytic activity.

[069] Most preferred is a mixture of transgalacto-oligosaccharide with an average DP below 10, preferably below 6, and a fructo-oligosaccharide with an average DP above 7, preferably above 11, even more preferably above 20. Such a mixture synergistically reduces protein fermentation in the colon and proteolytic activity.

[070] The present nutritional composition preferably comprises 0.5 to 20% by weight of total non-digestible oligosaccharide, more preferably 1 to 10% by weight, even more preferably 2 to 10% by weight, most preferably 2.0 to 7.5% by weight, based on the dry weight of the present composition.

[071] Based on 100 ml, the present enteric composition preferably comprises 0.1 to 2.5% by weight of total non-digestible oligosaccharide, more preferably 0.2 to 1.5% by weight, even more preferably 0.4 to 1.5% by weight, based on 100 ml of the present composition. NUTRITIONAL COMPOSITION

[072] The present nutritional composition is preferably particularly suitable for providing the complete daily nutritional requirements for a human individual under 36 months of age, more preferably a human infant. The present nutritional composition is not a yogurt, since yogurt conventionally contains L. bulgaricus Petition 870260046869, dated 05 / 18 / 2026, page 41 / 87 31 / 57 (Codex Standard for fermented milks Codex Stan 243-2003).

[073] The present nutritional composition comprises a digestible carbohydrate component. Preferred digestible carbohydrate components are lactose, glucose, sucrose, fructose, galactose, maltose, starch, and maltodextrin. Lactose is the main digestible carbohydrate present in human milk. The present nutritional composition preferably comprises lactose. As the present nutritional composition comprises a fermented composition comprising protein by lactic acid bacteria, the amount of lactose is reduced compared to its source due to fermentation whereby lactose is converted into lactate and / or lactic acid. Therefore, in the preparation of the present nutritional composition, lactose is preferably added. Preferably, the present nutritional composition does not comprise large amounts of carbohydrates other than lactose.Compared to digestible carbohydrates such as maltodextrin, sucrose, glucose, maltose, and other digestible carbohydrates with a high glycemic index, lactose has a lower glycemic index and is therefore preferred. The present nutritional composition preferably comprises digestible carbohydrate, of which at least 35% by weight, more preferably at least 50% by weight, more preferably at least 60% by weight, more preferably at least 75% by weight, even more preferably at least 9% by weight, more Preferably, at least 95% by weight of the digestible carbohydrate is lactose. Based on dry weight, the present nutritional composition preferably comprises at least 25% by weight of lactose, preferably at least 40% of which is lactose. Petition 870260046869, dated 05 / 18 / 2026, page 42 / 87 32 / 57 by weight, more preferably at least 50% by weight of lactose.

[074] Advantageously, the present nutritional composition is relatively low in protein and relatively high in lactose. Thus, in one embodiment, preferably the present nutritional composition comprises 1.6 to 2.1 g of protein per 100 kcal of the nutritional composition, preferably 1.6 to 2.0 g of protein per 100 kcal of the nutritional composition and the digestible carbohydrate component comprises at least 60% by weight of lactose based on total digestible carbohydrate, more preferably at least 75% by weight, even more preferably at least 90% by weight of lactose based on total digestible carbohydrate.

[075] When in liquid form, for example, as a ready-to-feed liquid, the nutritional composition preferably comprises 3.0 to 30 g of digestible carbohydrate per 100 ml, more preferably 6.0 to 20 g, even more preferably 7.0 to 10.0 g per 100 ml. Based on dry weight, the present nutritional composition preferably comprises 20 to 80% by weight, more preferably 40 to 65% by weight of digestible carbohydrates. Based on total calories, the nutritional composition comprises 5 to 20 g of digestible carbohydrates per 100 kcal, more preferably 8 to 15 g.

[076] The present nutritional composition comprises a lipid component. The lipid component of the present nutritional composition provides 3 to 7 g per 100 kcal of the nutritional composition, preferably the lipid component provides 4 to 6 g per 100 kcal. When in liquid form, for example, as a ready-to-drink liquid, the Petition 870260046869, dated 05 / 18 / 2026, page 43 / 87 33 / 57 The nutritional composition preferably comprises 2.1 to 6.5 g of lipid per 100 ml, more preferably 3.0 to 4.0 g per 100 ml. Based on dry weight, the present nutritional composition preferably comprises 12.5 to 40% by weight of lipid, more preferably 19 to 30% by weight.

[077] Preferably, the lipid component comprises the essential fatty acids alpha-linolenic acid (ALA), linoleic acid (LA) and / or long-chain polyunsaturated fatty acids (LC-PUFA). The LC-PUFA, LA and / or ALA may be supplied as free fatty acids, in triglyceride form, in diglyceride form, in monoglyceride form, in phospholipid form or as a mixture of one or more of the above. Preferably, the present nutritional composition contains at least one, preferably at least two lipid sources selected from the group consisting of canola oil (such as rapeseed oil, low erucic acid canola oil and canola oil), high oleic sunflower oil, high oleic safflower oil, olive oil, marine oils, microbial oils, EC coconut oil, palm kernel oil and milk fat.

[078] The present nutritional composition is not breast milk. The present nutritional composition comprises a lipid component, a protein component, and a digestible carbohydrate component. The nutritional composition according to the invention, or the nutritional composition used according to the invention, preferably comprises other fractions, such as vitamins, minerals, trace elements, and other micronutrients in order to make it a complete nutritional composition. Preferably, the nutritional composition is selected from the group consisting of a formula Petition 870260046869, dated 05 / 18 / 2026, page 44 / 87 34 / 57 Infant, infant milk or formula and growing-up milk, more preferably from the group consisting of an infant formula. An infant formula is defined as a formula for use in infants and may, for example, be a starter formula, for infants from 0 to 4 to 6 months of age, or as a follow-up formula, for infants from 4 to 6 months up to 12 months of age. An infant milk or growing-up milk or formula is for children from 12 to 36 months of age. In one embodiment, the nutritional composition is that of an infant formula. Infant formulas comprise vitamins, minerals, trace elements and other micronutrients in accordance with international guidelines.

[079] The lipid component provides 3 to 7 g of lipid per 100 kcal, preferably 4 to 6 g per 100 kcal, the protein component provides 1.6 to 4 g per 100 kcal, preferably 1.75 to 2.5 g per 100 kcal, and the digestible carbohydrate component provides 5 to 20 g per 100 kcal, preferably 8 to 15 g per 100 kcal of the final nutritional composition. Preferably, the present nutritional composition comprises a lipid component providing 4 to 6 g per 100 kcal, a protein component providing 1.6 to 1.9 g per 100 kcal, more preferably 1.75 to 1.85 g per 100 kcal, and a digestible carbohydrate component providing 8 to 15 g per 100 kcal of the final nutritional composition.In one embodiment, the lipid component provides 3 to 7 g of lipid per 100 kcal, preferably 4 to 6 g per 100 kcal, the protein component provides 1.6 to 2.1 g per 100 kcal, preferably 1.6 to 2.0 g per 100 kcal, and the digestible carbohydrate component provides 5 to 20 g per 100 kcal, preferably 8 to 15 g per 100 kcal of the final nutritional composition, wherein, preferably, the component of... Petition 870260046869, dated 05 / 18 / 2026, page 45 / 87 35 / 57 digestible carbohydrate comprises at least 60% by weight of lactose based on total digestible carbohydrate, more preferably at least 75% by weight, even more preferably at least 90% by weight of lactose based on total digestible carbohydrate.

[080] The total calorie count is determined by the sum of calories derived from protein, lipids, digestible carbohydrates, and non-digestible oligosaccharides.

[081] In one embodiment, the nutritional composition is in a liquid form. In another embodiment, the nutritional composition is a powder suitable for making a liquid nutritional composition after reconstitution with an aqueous solution, preferably with water. Preferably, the infant or child formula is a powder to be reconstituted with water. Preferably, the liquid composition has a viscosity below 100 mPa.s, more preferably below 60 mPa.s, more preferably below 35 mPa.s, even more preferably below 6 mPa.s as measured on a Brookfield viscometer at 20°C at a shear rate of 100 s-1. A low viscosity is important for infant or follow-up formula, as it mimics the viscosity of breast milk and can then be administered via a breast.

[082] In order to meet the caloric requirements of an infant or child, the nutritional composition preferably comprises 45 to 200 kcal / 100 ml of liquid. For infants, the nutritional composition is more preferably 60 to 90 kcal / 100 ml of liquid, even more preferably 65 to 75 kcal / 100 ml of liquid. This Petition 870260046869, dated 05 / 18 / 2026, page 46 / 87 A 36 / 57 caloric density ensures an ideal ratio between water and caloric intake. For children, human individuals aged between 12 and 36 months, the nutritional composition most preferably has a caloric density between 45 and 65, even more preferably between 50 and 60 kcal / 100 ml. The osmolarity of the present composition is preferably between 150 and 420 mOsmol / l, more preferably 260 to 320 mOsmol / l. The low osmolarity also aims to reduce gastrointestinal stress.

[083] When the nutritional composition is in a liquid form, the preferred volume administered on a daily basis is in the range of about 80 to 2500 ml, more preferably about 200 to 1200 ml per day. Preferably, the number of feedings per day is between 1 and 10, preferably between 3 and 8. In one embodiment, the nutritional composition is administered daily for a period of at least 2 days, preferably for one day for at least 4 weeks, preferably for a period of at least 8 weeks, more preferably for a period of at least 12 weeks, in a liquid form in which the total volume administered daily is between 200 ml and 1200 ml and in which the number of feedings per day is between 1 and 10. APPLICATION

[084] In the context of the present invention, 'prevention' of a disease or disorder also means 'reduction of the risk' of a disease or disorder and also means 'treatment of a person at risk' of said disease or disorder.

[085] The inventors discovered that by consuming the nutritional composition of the present invention the Petition 870260046869, dated 05 / 18 / 2026, page 47 / 87 37 / 57 The amount of endogenous proteolytic enzymes detected in the terminal ileum was significantly reduced compared to the amount detected by consuming a standard, unfermented ingredient containing the nutritional composition. Furthermore, despite a higher daily protein intake, apparent and true protein digestibility was higher in the group consuming the nutritional composition of the present invention.

[086] The nutritional composition of the present invention has been found to reduce digestive effort. Digestive effort is defined as the amount of protease activity secreted per gram of protein ingested.

[087] The nutritional composition of the present invention has been found to improve the efficiency of protein digestion. Efficiency of protein digestion is defined as the amount of protein ingested per arbitrary unit (AU) of protease activity. The nutritional composition of the present invention has been found to reduce the loss of endogenous protein, such as the loss of endogenously formed proteases. This was, in particular, the case for trypsin and / or chymotrypsin and / or elastase. Trypsin and chymotrypsin are, in an infant, the most important digestive proteases, since gastric pepsin is less active due to a higher pH in the stomach. Furthermore, proteases have been found to stimulate the endogenous loss of other proteins, such as mucins.

[088] It has been found that the nutritional composition of the present invention reduces the protein load that enters the colon.

[089] Thus, through feeding, the nutritional composition of the present invention, the release of protease Petition 870260046869, dated 05 / 18 / 2026, page 48 / 87 38 / 57 endogenous, digestive effort, loss of endogenous protein in the small intestine and the protein load entering the colon are advantageously reduced, as well as the proteolytic activity of the microbiota in the colon. The nutritional composition of the present invention will have an enhanced effect on reducing protein fermentation and proteolytic activity in the colon and will have a beneficial effect with respect to reducing proteolytic activity throughout the entire intestinal tract, i.e., small and large intestine.

[090] Therefore, the nutritional composition of the present invention is advantageously used as nutrition for infants and children, for use in the prevention and / or treatment of diaper rash or in promoting intestinal tract health by reducing digestive effort, improving or increasing the effectiveness of protein digestion, reducing endogenous protein loss, reducing endogenous protease secretion, reducing colon protein fermentation and / or reducing the protein load entering the colon.

[091] The discovery that the efficiency of protein digestion is increased, and endogenous protein loss is decreased, also advantageously allows the formulation of an infant or child formula with lower protein concentrations than those used until now.

[092] The effects described here are observed compared to the situation before administration of the nutritional composition and / or to the situation compared to administration of a standard nutritional composition not comprising the fermented composition comprising protein and non-digestible oligosaccharides. It was found that these observed effects also approximate the effects that occur in breastfed infants compared to breastfed infants. Petition 870260046869, dated 05 / 18 / 2026, page 49 / 87 39 / 57 fed standard infant formula, as proteolytic activity was found in fecal samples from exclusively breastfed infants compared to those fed standard formula.

[093] Due to the effects mentioned above, the nutritional composition of the present invention improves intestinal tract health. In one embodiment, the present nutritional composition for infants is for use in promoting intestinal tract health. Preferably, intestinal tract health is selected from the group consisting of i) increased intestinal barrier function and ii) use in the prevention and / or treatment of a disorder selected from the group consisting of constipation, abdominal pain, abdominal discomfort, colic, cramps, abdominal bloating, abdominal distension, irritable bowel syndrome, and intestinal inflammation. In one embodiment, intestinal tract health is selected from the group consisting of inflammatory bowel disease (IBD) such as ulcerative colitis and diarrheagenic irritable bowel syndrome (IBS-D).

[094] Preferably, intestinal tract health is selected from the group consisting of constipation, abdominal pain, abdominal discomfort, cramps, abdominal bloating, abdominal distension, and irritable bowel syndrome. In one embodiment, intestinal tract health is selected from intestinal inflammation. In one embodiment, the nutritional composition of the present invention improves intestinal tract health by preventing and / or treating constipation. Constipation can be assessed or described as a decreased frequency of defecation and / or an increase in stool consistency. Petition 870260046869, dated 05 / 18 / 2026, page 50 / 87 40 / 57

[095] In particular, the reduced amount of protein entering the colon will result in more saccharolytic and less proteolytic activity of the gut microbiota. The fermentation of sugars, rather than amino acids, will result in a lower colon pH and / or reduced formation of toxic metabolites such as indoles, phenols, and amines. This will also result in more Bifidobacteria and / or Lactobacilli and / or fewer pathogenic bacteria in the gut microbiota. Bacterial quantities can be expressed as cfu per g of feces and / or as a cfu-based percentage of total bacteria. Such an improved gut microbiota will result in reduced intestinal infections and / or reduced diarrhea.

[096] In particular, the reduced amount of proteases, more specifically serine proteases such as trypsin and chymotrypsin, will result in reduced PAR-2 ​​cleavage with the effect of an increased intestinal barrier function. An increased barrier function will result in reduced translocation of toxins, allergens and pathogens and, therefore, in an advantageous effect on infection, diarrhea and / or inflammation. Also, reduced PAR-2 ​​cleavage will result in less perception of abdominal pain. Increased fecal proteolytic activity is, in particular, associated with inflammatory bowel disease (IBD) such as ulcerative colitis and irritable bowel syndrome diarrhea (IBS-D). Thus, in one embodiment, the nutritional composition of the present invention is used for the treatment and / or prevention of inflammatory bowel disease such as ulcerative colitis and irritable bowel syndrome diarrhea (IBS-D).Increased fecal proteolytic activity is particularly associated with the occurrence of diaper rash. Petition 870260046869, dated 05 / 18 / 2026, page 51 / 87 41 / 57 Therefore, the present nutritional composition of the invention is preferably for use in the treatment and / or prevention of diaper rash. Furthermore, lower protease synthesis, concomitant with increased protein digestibility, results in energy and protein loss, which improves growth and development.

[097] Colonic fermentation of proteins will result in changes in the quality or quantity of gas formation and, therefore, increased abdominal swelling and / or abdominal distension and / or flatulence. Therefore, the present nutritional composition will have an effect on reducing abdominal swelling and / or abdominal distension and disorders resulting therefrom. Activation of protease-activated receptors increases color perception and has a negative effect on intestinal barrier function. Lower proteolytic activity may thus contribute to the prevention and / or enhanced treatment of colic or cramps. Cramps can be assessed when a baby arches their back, and colic can be assessed when there is excessive crying time. In one embodiment, the nutritional composition of the present invention is used for the treatment and / or prevention of colic and / or cramps.

[098] All of these effects mentioned and observed above are particularly important in young human individuals, since they need to grow and develop, have an immature intestinal barrier, and a less developed gut microbiota. In other words, in young human individuals, improving the digestive effort of protein, regulating the release of endogenous protein, restricting protein loss, and decreasing the protein load in the colon is Petition 870260046869, dated 05 / 18 / 2026, page 52 / 87 42 / 57 most important. The development of the intestinal tract is a complex process. It is plausible that leakage of the intestinal tract makes babies during the first months of life more susceptible to elevated levels of proteolytic activity, which can trigger low levels of inflammation and increased pain perception. This may explain the beneficial effects of the present nutritional composition that induces the release of less endogenous protease. Therefore, the nutritional composition is preferentially used to feed a baby.

[099] In one embodiment, the invention relates to a method, preferably a non-therapeutic method, for a) to increase the efficiency of protein digestion, b) reduce the secretion of proteases in the small and / or large intestine, c) reduce proteolytic activity in the small and / or large intestine, d) reduce the protein load entering the large intestine and / or (e) reducing protein fermentation in the large intestine in a human subject aged 0 to 36 months, the method comprising administering a nutritional composition according to the invention to the human subject aged 0 to 36 months, wherein the increase or decrease is compared to the effect of a nutritional composition not comprising a fermented composition comprising protein by lactic acid bacteria and non-digestible oligosaccharides. Preferably for this method, the nutritional composition includes lactose.

[0100] In one embodiment, the protease is a serine Petition 870260046869, dated 05 / 18 / 2026, page 53 / 87 43 / 57 protease and proteolytic activity is the proteolytic activity of a serine protease, more preferably a serine protease selected from the group consisting of trypsin, chymotrypsin, and elastase.

[0101] The present nutritional composition is used to provide nutrition for a human individual aged 0 to 36 months. In one embodiment, the present nutritional composition is used to provide nutrition for an infant aged 0 to 18 months, more preferably an infant aged 12 months or younger. In another embodiment, the present nutritional composition is used to provide nutrition for a child aged 18 to 36 months, more preferably a child aged 18 to 30, or 24 months. The present nutritional compositions are enterically administered, more preferably orally.

[0102] In this document and its claims, the verb “to include” and its conjugations are used in their non-limiting sense to mean that the items following the word are included, but items not specifically mentioned are not excluded. Furthermore, reference to an element by the indefinite article “a” or “an” does not exclude the possibility that more than one of the elements is present, unless the context clearly requires that there be more than one and only one of the elements. The indefinite article “a” or “an” normally means “at least one”. Wt. means weight. EXAMPLES EXAMPLE 1

[0103] A double-blind, placebo-controlled, randomized, prospective study with a parallel-group design was conducted in healthy newborns up to the age of one Petition 870260046869, dated 05 / 18 / 2026, p. 54 / 87 44 / 57 year of the baby. The test groups were: Group 1: Infants exclusively breastfed for at least 4 months (N=43). Group 2: Infants exclusively formula-fed after a maximum of 2 months, receiving a standard non-hydrolyzed cow's milk-based formula (N=41). Group 3: Infants exclusively formula-fed after 2 months of age, receiving a standard non-hydrolyzed cow's milk-based formula containing a mixture of non-digestible carbohydrates (0.8 g / 100 ml) of neutral and acidic oligosaccharides in a weight ratio of 85:15. As neutral oligosaccharides, transgalactooligosaccharides (source VivinalGOS) and long-chain oligosaccharides (source raftiniln HP) in a weight ratio of 9:1 were used. As acidic oligosaccharides, uronic acid oligosaccharides derived from pectin degradation were used. (N=36).

[0104] Stool samples were collected at week 16 after birth and stored at -20 °C until further analysis. Fecal proteolytic activity in fecal supernatant was measured using the EnzCheck protease fluorescence assay kit (E6638, Invitrogen, Carlsbad, CA, USA) to detect metallo-, serine, and sulfhydryl proteases.Fecal samples were diluted 10x in 1x PBS, homogenized for 5 min, and centrifuged at 13,000 rpm for 3 minutes to remove large particles. 100 µl (100x diluted) of supernatant were added to 100 µl of working BODIPY casein (10 pg BODIPY casein). Fluorescence increase was measured at 25°C for 10 minutes. Porcine pancreatin (Sigma, P1750) was used to prepare a calibration curve. Petition 870260046869, dated 05 / 18 / 2026, page 55 / 87 45 / 57 To determine the origin of protease activity, the specific serine protease inhibitor AEBSF (Sigma, A8456) was added to each analysis at a final concentration of 5 mM. Total protein in fecal supernatants was determined using the BCA method (Pierce). Fecal proteolytic activity was expressed as arbitrary units (AU) based on pancreatin activity in USP (standard unit for activity according to the United States Pharmacopeia) per milligram of fecal protein content. Statistical analysis was performed using the Mann-Whitney U test. RESULTS

[0105] The results are shown in Table 1. Fecal proteolytic activity in the standard infant formula group was significantly higher compared to the breastfed group and the group on standard infant formula containing the oligosaccharide mixture. Fecal proteolytic activity was similar between the groups on breast milk and standard infant formula containing the non-digestible oligosaccharide mixture.

[0106] Table 1: Mean proteolytic activity (AU / mg of protein) in the stool of infants fed breast milk (group 1), a control infant formula (group 2) or a formula with non-digestible carbohydrates (Group 3). Diet Proteolytic activity (SE) P-value * v group 1 P-value * v group 2 Group 1 313 (21) 0.032 Group 2 406 (33) 0.032 Group 3 309 (26) 0.488 0.020 *: Mann-Whitney U test.

[0107] The proteolytic activity was mainly derived from serine proteases (i.e., enzymes Petition 870260046869, dated 05 / 18 / 2026, pp. 56 / 87 46 / 57 pancreatic or produced by colon microbiota), since the specific serine protease inhibitor AEBSF blocked proteolytic activity by more than 70%.

[0108] These experiments are indicative of reduced proteolytic activity in the colon and reduced protein fermentation in the presence of non-digestible oligosaccharides. EXAMPLE 2: EXPERIMENTS WITH PIGS AND FERMENTED INFANT FORMULA

[0109] To investigate protein digestion in vivo, pigs are first chosen because they closely resemble human digestive physiology (Miller et al. 1987, Annu Rev Nutr, 7:361-82) and generate comparable true ileal nitrogen and amino acid (AA) digestibilities (Deglaire et al. 2009, Br J Nutr, 102(12):1752-9, Rowan et al. 1994, Br J Nutr, 71(1):29-42). Furthermore, a study by Moughan et al. showed that three-week-old piglets can be used as a model for 6-month-old infants (Moughan et al. 1991, J Nutr, 121(10):1570-4). Therefore, the AA digestibility of a standard and fermented infant formula (IF) was tested in a piglet model. A hydrolyzed formula (Nutrilon Pepti) was used to correct endogenous AA losses according to a peptide feeding method (Rutherfurd et al. 1998, J Dairy Sci, 81(4):909-17).

[0110] Six male guinea pigs of corresponding age and weight (average weight 4.9 kg) were housed in groups at 2 weeks of age. At three weeks of age, they received a T-cannula in the distal part of the terminal ileum after overnight fasting and were housed individually in Petition 870260046869, dated 05 / 18 / 2026, pp. 57 / 87 47 / 57 followed. From day 31 onwards, the pigs received the following diets: Diet 1: Lactofidus-1, a commercially available infant milk formula marketed under the trade name Gallia, comprising 100% of a composition derived from fermented milk. Lactofidus is produced by fermentation with S. thermophilus and comprising B. breve. A heat treatment is employed to inactivate the lactic acid bacteria. It contains 1.1% by weight of lactic acid + lactate based on dry weight, which is at least 95% lactic acid / lactate. It comprises 12.2 g of protein per 100 g dry weight, of which 60% by weight is casein and 40% by weight is whey protein. The pH is 5.6. Diet 2: Nutrilon-2, a standard non-fermented follow-up formula under the trade name Nutricia, comprising 9.3 g of non-hydrolyzed protein per 100 g dry weight, of which 50% by weight is non-hydrolyzed whey protein and 50% by weight is casein. The pH is 6.8. Diet 3: Pepti-2, a marketed IMF under the trade name Nutricia comprising 11.2 g of protein per 100 g dry weight in the form of 100% by weight extensively hydrolyzed whey protein. The pH is 6.4.

[0111] Standard fermented, hydrolyzed IMF powder was diluted in demineralized water (37 °C) to a final dry matter content of 21.0%, 22.05%, and 21.45%, respectively (protein content being 2.46, 2.05, and 2.55 g / 100 ml). Chromium oxide was added to the IMF powder as a non-digestible marker. Until day 31, the guinea pigs received a 1:1:1 w / w / w mixture of diets 1, 2, and 3.

[0112] From day 31 to day 36, the little pigs Petition 870260046869, dated 05 / 18 / 2026, p. 58 / 87 48 / 57 received diet 1, 2, or 3 in a Latin Square design according to a feeding schedule from Table 1. Pigs were fed at 7:00 h, 9:30 h, 14:00 h, and 16:00 h. Ileal digesta was collected on days 32, 34, and 36 from 8:00 h to 17:00 h, using a T-cannula in small bags that were stored on ice immediately upon filling. All digesta samples were weighed, pH measured, and stored at -20 °C until further processing. For enzymatic activity measurements, a subsample (2 ml) was centrifuged (13,000 g for 10 minutes at 4 °C), and supernatants were aliquoted and stored at -80 °C. TABLE 2: POWER SUPPLY SCHEME Little Pig Day 31-32 Day 33-34 Day 35-36 1 1 2 3 2 1 3 2 3 2 1 3 4 2 3 1 5 3 1 2 6 3 2 1

[0113] Dry matter (DM) (gravimetry at 80 °C), chromium oxide (Cr) (inductively coupled plasma mass spectrometry), crude protein (CP) (Kjeldahl method, N x 6.25) and amino acid composition (AA) (HPLC after 6M HCl hydrolysis) were analyzed in freeze-dried digesta samples and diet powders.

[0114] Total proteolytic activity in ileal digesta was determined using the EnzCheck protease fluorescence-based assay kit (E6638, Invitrogen, Carlsbad, CA, USA) to detect metallo-, serine, and sulfhydryl proteases. Digesta were diluted 750x with 10 mM Tris-HCl, Petition 870260046869, dated 05 / 18 / 2026, page 59 / 87 49 / 57 pH 7.8, and 100 μl of samples were added to 100 μl of working BODIPY casein (10 μg BODIPY casein). The increase in fluorescence was measured at 25°C for a period of 10 minutes. Porcine pancreatin (Sigma, P1750) was used to prepare a calibration curve. To determine the origin of protease activity, the specific serine protease inhibitor AEBSF (Sigma, A8456) was added to each analysis with a final concentration of 5 mM. Activity is expressed as an arbitrary unit (AU) (based on USP pancreatin activity).

[0115] Trypsin activity was measured using Na-Benzoyl-L-Arginine Ethyl Ester (BAEE, Sigma B4500) as a substrate and signifying the change in absorbance at 25°C at 253 nm (according to the manufacturer's instructions). Bovine trypsin (Sigma, T9201) was used to prepare a calibration curve. Activity is expressed as an arbitrary unit (AU) (based on trypsin activity in U).

[0116] Chymotrypsin was measured using Ethyl Na-Benzoyl-L-Tyrosine ester (BTEE, Sigma B6125) was used as a substrate, signifying the change in absorbance at 25°C at 256 nm. Bovine chymotrypsin (Sigma, C3142) was used to prepare a calibration curve. Activity is expressed as an arbitrary unit (AU) (based on chymotrypsin activity in U).

[0117] Elastase activity was measured using SucAla3-PNA (S4760, Sigma) as a substrate and porcine elastase (E7885, Sigma) was used to prepare a calibration curve. Activity is expressed as an arbitrary unit (AU) (based on elastase activity in U).

[0118] Equations used to calculate AA digestibility (units are in μg / g of DMI): Petition 870260046869, dated 05 / 18 / 2026, pp. 60 / 87 50 / 57 Flow AA flow Dietary creatinine of ileal amino acids = ileal amino acids digested -----Digested creatinine Endogenous dietary creatinine = Ileal amino acids (MW>10 kD) x Digested creatinine Apparent ileal amino acid digestibility (%) AA intake - ileal AA flow AA intake x 100% Digestibility of amino acids in the true ileum (%) = AA intake - (ileal AA flow - endogenous AA flow) AA intake 100%

[0119] The results were analyzed using Univariate Analysis of Variation (GLM procedure). Differences between diets were considered significant with p<0.05 according to an LSD test.

[0120] The results in Table 3 show that the pH of the ileal digesta was the same. The osmolarity was also the same. The amount of crude protein intake was higher in the hydrolyzed formula and the fermented formula. Interestingly, the amount of total protein in the ileal digesta and the relative amount of protein based on protein intake was significantly lower in guinea pigs that consumed fermented formula than in the other two groups. This protein flow to the colon is therefore lower in guinea pigs that consume fermented formula. The apparent and true ileal crude protein digestibility was higher in guinea pigs that consumed fermented formula than in the group that consumed standard formula. The apparent and true ileal amino acid digestibility was higher in guinea pigs that consumed fermented formula than in the group that consumed standard formula. The same applies to most individual amino acids. The digestibility of... Petition 870260046869, dated 05 / 18 / 2026, pp. 61 / 87 51 / 57 true glycine differs most notably. This is indicative of a decrease in endogenous protein secretion. The endogenous amino acid flux was primarily rich in glutamic acid, threonine, aspartic acid, proline, and serine. TABLE 3. DIETARY INTAKE, CHARACTERISTICS OF Ileal Digestion & Ileal Amino Acid Digestibility Nutrilon Lactofidus Pepti Pooled SE DM Intake (g / day) 403a 388b 391b 3.641 CP Intake (g / day) 37.4a 45.4b 46.4c 0.317 Total AA Intake (g / day) 39.1a 47.7b 50.2c 0.347 Ileal digesta osmolarity (mOsm / kg) 305 305 309 3.3 Ileal digesta pH 7.91 7.95 7.90 0.06 Ileal digesta CP (g / g CPI) 0.16a 0.08b 0.16a 0.01 Total ileal digesta CP (g / day) 5.96a 3.64b 7.27c 0.279 Apparent ileal crude protein digestibility (%) 84.2a 92.0bx 0.81 Apparent ileal amino acid digestibility (%) 89.1a 94.4bx 0.53 Alanine 85.0a 91.1bx 0.81 Arginine 85.8a 93.1bx 0.68 Aspartic acid 88.6a 93.8bx 0.51 Cystine 85.1 89.5 x 1.06 Glutamic acid 93.1a 9 6.3bx 0.29 Petition 870260046869, dated 05 / 18 / 2026, p. 62 / 87 52 / 57 Glycine 48.5 7 8.5bx 4.81 Histidine 88.9a 94.5bx 0.51 Iso-leucine 91.9a 95.8bx 0.34 Leucine 93.4a 9 6.5bx 0.29 Lysine 91.1a 96.1bx 0.31 Methionine 93.9a 97.2bx 0.36 Phenylalanine 88.8a 94.6bx 0.40 Proline 90.1a 95.4bx 0.59 Serine 84.6a 92.2bx 0.63 Threonine 79.0a 88.1bx 1.05 Tyrosine 90.7a 94.7bx 0.37 Valine 90.6a 95.2bx 0.54 Digestibility of crude protein from true ileum (%) 94.2a 9 9.9bx 0.89 Digestibility of amino acids from true ileum (%) 97.1a 100.8bx 0.61 Alanine 96.7a 101.2bx 0.87 Arginine 94.4a 99.4bx 0.82 Aspartic acid 97.3a 101.3bx 0.58 Cystine 97.0a 102.2bx 1.01 Glutamic acid 98.6a 100.6bx 0.34 Glycine 72.4a 97.4bx 4.74 Histidine 96.4a 100.3bx 0.63 Isoleucine 99.1a 101.7bx 0.42 Leucine 98.5a 100.7bx 0.35 Lysine 96.6a 100.6bx 0.37 Methionine 98.8a 100.9bx 0.42 Phenylalanine 96.3a 100.1bx 0.53 Proline 97.4a 100.8bx 0.66 Petition 870260046869, dated 05 / 18 / 2026, pp. 63 / 87 53 / 57 Serine 96.6a 101.3bx 0.73 Threonine 96.5a 102.7bx 1.12 Tyrosine 97.8 100.0 x 0.53 Valine 98.5a 101.3bx 0.43 x = undetermined. (ac) Values ​​with different letters within the same line are different (p<0.05) TABLE 4. PROTEOLYTIC ENZYME ACTIVITIES OF THE ILEUM Nutrilon Lactofidus Pepti Pooled SE Total proteolytic activity AU x 103 / 8h 1599a 477b 759b 140 AU x 103 / g CPI 41.4a 10.5b 16.5b 3.3 Trypsin activity AU x 103 / 8h 908a 334b ​​621c 74 AU / g CPI 23, 9a 7.4b 13.5c 1.6 Chymotrypsin activity AU / 8h 665a 294b 4 8 6ab 73 AU / g CPI 17.2a 6.5b 10.3b 1.8 Elastase activity AU / 8h 191a 6 9b 123c 13 AU / CPI 4.78a 1.45b 2.47c 0.27 NS = not significant, x = not determined. (ac) Values ​​with different letters within the same line are different (p<0.05)

[0121] The results in Table 4 show that the amount of proteolytic activity secreted during the 8 h of collection is lowest in guinea pigs that consume Lactofidus, the fermented infant formula, when compared with standard non-fermented formula or a hydrolyzed, pre-digested IMF. Petition 870260046869, dated 05 / 18 / 2026, pp. 64 / 87 54 / 57 This is the case for total proteolytic activity as well as for trypsin and chymotrypsin. This is also the case for elastase activity. This effect is also observed when based on the amount of protein intake.

[0122] Proteolytic activity is mainly derived from serine proteases (i.e., pancreatic enzymes such as trypsin and chymotrypsin), since the specific serine protease inhibitor AEBSF blocked proteolytic activity by more than 90%. Thus, the amount of proteolytic activity that enters the colon is lower when consuming a fermented formula, making it more similar to the situation in breastfed infants (see example 1).

[0123] Surprisingly, apparent and true protein digestibility is highest when consuming the fermented formula, despite the lower amount of proteases being formed. This means that digestive efficiency (in [protein ingested / protease activity in AU]) is higher. Similarly, the amount of proteolytic activity per g of protein intake is lower, which is indicative of reduced digestive effort. EXAMPLE 3: LOW-PROTEIN INFANT FORMULA

[0124] A powdered infant formula which, after reconstitution with water, becomes a ready-to-feed liquid infant formula comprising 100 ml: Approximately 13.0 g of dry matter, 66 kcal 1.2 g of protein (bovine whey protein / casein in a weight ratio of 1 / 1), 9.8% by weight based on dry weight, 1.8 g per 100 kcal. 7.3 g of digestible carbohydrate (7.3 g of sugars, of which 7.1 g are lactose) (11.4 g / 100 kcal). Petition 870260046869, dated 05 / 18 / 2026, pp. 65 / 87 55 / 57 3.36 g of fat (mostly vegetable fat). (50.9 g / 100 kcal) 0.8 g of non-digestible oligosaccharides from scGOS (source Vivinal GOS) and lcFOS (source RaftilinHP) in a weight ratio of 9:1, of which 0.6 g is classified as dietary fiber and 0.2 g is non-digestible saccharides present in scGOS, which is classified as carbohydrates.

[0125] Of this composition, 30% on a dry weight basis are derived from lactofidus-1 as described in example 2. The composition comprises approximately 0.33% by weight of lactic acid + lactate on a dry weight basis, of which at least 95% is L-lactate / lactic acid. The composition further comprises vitamins, minerals, trace elements and other micronutrients in accordance with international guidelines for infant formula.

[0126] The pH after reconstitution is 6.2. EXAMPLE 4: INFANT FORMULA

[0127] A powdered infant formula which, after reconstitution with water, becomes a ready-to-feed liquid infant formula comprising 100 ml: Approximately 13.7 g of dry matter, 66 kcal 1.35 g of protein (bovine whey protein / casein in a 1 / 1 weight ratio), 11% by weight based on dry weight, 2.0 g / 100 kcal 8.2 g of digestible carbohydrate (of which 5.6 g is lactose and 2.1 g is maltodextrin) 3.0 g of fat (mostly vegetable fat). 0.8 g of non-digestible oligosaccharides of scGOS (source Vivinal GOS) and lcFOS (source RaftilinHP) in a weight ratio of 9:1. Petition 870260046869, dated 05 / 18 / 2026, pp. 66 / 87 56 / 57

[0128] Of this composition, 50% on a dry weight basis are derived from lactofidus-1 as described in example 2. The composition comprises approximately 0.55% by weight of lactic acid + lactate on a dry weight basis, of which at least 95% is L-lactic acid / lactate. The composition further comprises vitamins, minerals, trace elements and other micronutrients in accordance with international directive 2006 / 141 / EC for infant formula.

[0129] Infant formula is intended to promote intestinal tract health and / or increase intestinal barrier function and / or prevent and / or treat constipation, abdominal pain, abdominal discomfort, colic, cramps, abdominal bloating, abdominal distension, irritable bowel syndrome, ulcerative colitis, diarrheagenic irritable bowel syndrome or intestinal inflammation, and / or prevent and / or treat diaper rash. EXAMPLE 5: FORMULA FOR CHILDREN

[0130] A ready-to-drink liquid for children aged 12 to 36 months, comprising per 100 ml: Approximately 56 kcal 1.5 g of protein (bovine whey protein / casein in a weight ratio of 1 / 1), 2.7 g / 100 kcal 6.6 g of digestible carbohydrate (of which more than 95% by weight is lactose) 1.94 g of fat (mostly vegetable fat). 1.2 g of non-digestible oligosaccharides of scGOS (source Vivinal GOS) and lcFOS (source RaftilinHP) in a weight ratio of 9:1.

[0131] Of this composition, 15% based on dry weight are derived from lactofidus-1 as described in example 2. Petition 870260046869, dated 05 / 18 / 2026, pp. 67 / 87 57 / 57 The composition comprises approximately 0.17% by weight of lactic acid + lactate based on dry weight, of which at least 95% is L-lactic acid / lactate. The composition further includes vitamins, minerals, trace elements and other micronutrients as known in the art.

[0132] The pH of this composition is 6.6.

Claims

1. NUTRITIONAL COMPOSITION for a human individual aged between 0 and 36 months, characterized by comprising protein, lipids, digestible carbohydrates and non-digestible oligosaccharides, wherein: protein is present in an amount of 5 to 20% by weight based on the dry weight of the nutritional composition and in an amount of 1.6 to 2.0 g per 100 kcal; lipids are present in an amount of 3 to 7 g per 100 kcal; digestible carbohydrates are present in an amount of 5 to 20 g per 100 kcal, wherein at least 95% by weight of the digestible carbohydrates are lactose; non-digestible oligosaccharides are present in an amount of 0.5 to 20% by weight based on the dry weight of the nutritional composition, wherein said nutritional composition comprises at least 10% by weight, based on the dry weight of the nutritional composition, of a protein comprising a composition fermented by bacteria of lactic acid, and whose nutritional composition comprises from 0.20 to 1,0% by weight of the sum of lactate and lactic acid based on the dry weight of the nutritional composition, and where the sum of L-lactic acid and L-lactate is more than 50% by weight based on the sum of total lactic acid and lactate.

2. NUTRITIONAL COMPOSITION, according to claim 1, characterized by the nutritional composition being a liquid having a pH of 5.5 to 7.

0.

3. NUTRITIONAL COMPOSITION, according to any of claims 1 or 2, characterized by the nutritional composition comprising digestible carbohydrate and in which at least 75% by weight of the digestible carbohydrate is lactose.

4. NUTRITIONAL COMPOSITION, according to any one of claims 1 to 3, characterized in that at least one, more preferably at least two, of the non-digestible oligosaccharides are selected from the group consisting of galacto-oligosaccharides, fructooligosaccharides, uronic acid oligosaccharides, glucooligosaccharides, xylo-oligosaccharides, mannan-oligosaccharides, arabino-oligosaccharides, glucomannouligosaccharides, galactomannouligosaccharides, soy oligosaccharides, isomalto-oligosaccharides, non-digestible dextrin, arabinogalacto-oligosaccharides, gentiooligosaccharides, nigero-oligosaccharides, glucomannouligosaccharides, chito-oligosaccharides, sialyloligosaccharides, and fuco-oligosaccharides.

5. NUTRITIONAL COMPOSITION, according to any one of claims 1 to 4, characterized by the non-digestible oligosaccharides being selected from the group consisting of transgalacto-oligosaccharides, fructooligosaccharides and galacturonic acid oligosaccharides and mixtures thereof.

6. NUTRITIONAL COMPOSITION, according to any one of claims 1 to 5, characterized by the nutritional composition comprising 102-105 colony-forming units (CFU) of live Streptococcus thermophilus per gram of dry weight of the nutritional composition.

7. USE OF THE NUTRITIONAL COMPOSITION, as defined in any of claims 1 to 6, characterized by being in the manufacture of an infant formula for babies from 0 to 36 months. Petition 870260046869, dated 05 / 18 / 2026, page 1.70 / 87 3 / 4 months to promote the intestinal tract health of said infants by: a) increasing the efficiency of protein digestion, b) reducing the secretion of proteases in the small and / or large intestine, c) reducing proteolytic activity in the small and / or large intestine, d) reducing the protein load entering the large intestine and / or e) reducing protein fermentation in the large intestine, wherein intestinal tract health is selected from the group consisting of i) increased intestinal barrier function and ii) prevention and / or treatment of a disorder selected from the group consisting of constipation, abdominal pain, abdominal discomfort, colic, cramps, abdominal bloating, abdominal distension caused by irritable bowel syndrome, ulcerative colitis, diarrheagenic irritable bowel syndrome and intestinal inflammation.

8. USE OF THE NUTRITIONAL COMPOSITION, according to claim 7, characterized in that the protease is a serine protease and the proteolytic activity is the proteolytic activity of a serine protease, more preferably a serine protease selected from the group consisting of trypsin, chymotrypsin and elastase.

9. USE OF THE NUTRITIONAL COMPOSITION, according to claim 7, characterized by being for the treatment and / or prevention of diaper rash, colic and / or cramps in said babies.

10. USE OF THE NUTRITIONAL COMPOSITION, as defined in Petition 870260046869, dated 05 / 18 / 2026, page 71 / 87 4 / 4 in any of claims 1 to 6, characterized as being for feeding an infant.