Infant formula, uses of infant formula, and methods to reduce protein digestibility.

Description

1 / 29 Infant formula, uses of infant formula, and methods to reduce protein digestibility. Split Order BR 11 2021 012174-9, filed on 12 / 19 / 2019 Field of Invention

[001] This invention is in the area of ​​protein compositions. In particular, the invention relates to protein compositions comprising proteins with a high isoelectric point, suitable for use in infant formulas, as well as infant milk formulas comprising the protein compositions of the invention and the use thereof to improve digestibility and / or prevent / treat conditions associated with suboptimal or impaired digestibility in infants and / or toddlers. Background of the Invention

[002] Human milk is the gold standard for infant and young child nutrition. However, breast milk is not always an available option. When this is the case, infant formula is the first choice for replacing human milk and supports the proper growth of infants when fed as the sole or combined source of nutrition. Human milk is a complex matrix of components with unique molecular structure and biological function that are not easily replicated in infant formula, especially considering the heat treatments the product must undergo to ensure its safety and stability through long-term storage.

[003] Although cow's milk is a valuable source of components for the infant formula industry, it differs considerably from human milk in terms of protein content and concentration. The complex structure of proteins and their interactions in human milk are believed to be intrinsically linked to the health benefits of human milk in infants, which is therefore difficult to replicate in infant formulas.

[004] Protein digestion occurs in two phases: gastric and intestinal. Petition 870260056360, dated 10 / 06 / 2026, page 20 / 84 2 / 29 In the stomach (pH around 6.5 to 3.0), pepsin partially breaks down proteins into large polypeptides and oligopeptides. After a certain period of time in the stomach, the gastric contents are emptied and proceed to the intestinal phase. In the intestine (pH around 6.6), trypsin and peptidases will hydrolyze the polypeptides into tripeptides, dipeptides, and free amino acids, which are small enough to be transported through the cells into the bloodstream.

[005] Ideally, the digestibility of infant formulas should be comparable to the digestibility of human milk, as the latter is the gold standard for human nutrition and development. However, it is known that digestibility in infants fed formula or cow's milk differs from that of infants fed human milk: the digestibility of human milk is considerably lower than the digestibility of currently available standard infant formulas, resulting in a faster postprandial appearance of amino acids in the blood with infant formula than with human milk (Moro et al. “Postprandial plasma amino acids in preterm infants: influence of the protein source” Act. Pediatr. 1999, 88(8):885 to 889).

[006] It is desirable to mimic the lower digestibility profile of human milk, as it is believed that lower protein digestibility promotes proper gastrointestinal tract development and nutrient absorption in infants. Slowly absorbed proteins promote postprandial protein deposition, while rapidly absorbed proteins, such as in commercially available standard milk formula, stimulate amino acid oxidation, ultimately affecting tissue development in infants. It is further believed that the lower digestibility of human milk in infants / children who are beginning to walk promotes the adequate supply of proteins, peptides, and amino acids to specific sections of the gastrointestinal tract, which are responsible for signaling the morphological and functional development of the aforementioned intestinal section. Furthermore, the consequences of suboptimal or deficient development of functions Petition 870260056360, dated 10 / 06 / 2026, p. 21 / 84 3 / 29 Intestinal or nutrient absorption problems in formula-fed infants are not only immediate, but can also persist into adulthood, creating a basis for increased susceptibility to a range of diseases and conditions, such as obesity, reduced immunity, etc.

[007] Attempts have been made to modulate the digestion of babies.

[008] WO 2012 / 045801 describes compositions containing a complex coacervate comprising lactoferrin and at least one other protein with an isoelectric point less than pH 7.0. Methods for preparing these protein-protein complexes are described. Complex coacervates are said to be stable and easy to incorporate into food products. In the examples, the preparation of the coacervates is described, and the in vitro digestion of protein solutions comprising the coacervates is tested.

[009] US Patent 6,190,724 B1 describes compositions comprising modified proteins and methods for modifying proteins. The methods include 1) polymerization and / or crosslinking, 2) inclusion of certain free amino acids, 3) glycosylation, 4) dephosphorylation, or 5) partial or total thermal rearrangement of proteins. The modified proteins are used in infant formulas in an amount of at least 15%, by weight, of the total protein content. The modified proteins are described as altering the kinetics of digestion (i.e., slowing it down).

[010] There is still a need to improve the digestibility of milk formulas for infants and young children and to have a nutrient absorption profile that mimics that of human milk. Brief Description of the Invention

[011] Protein compositions for standard infant formulas typically comprise casein and whey protein, such as cow's milk. Standard infant formulas have higher digestibility than breast milk, which is undesirable. Reference is made to Figure 1 in this application. Petition 870260056360, dated 10 / 06 / 2026, p. 22 / 84 4 / 29

[012] The inventors discovered that the inclusion of substantial amounts of protein(s) with a high isoelectric point (pI) in compositions comprising casein and whey protein surprisingly lowers the protein digestibility profile compared to compositions currently used in standard infant formulas, i.e., without these high pI proteins. As used in the present application, “low” / “lower” digestibility does not imply that the proteins are digested to a lesser degree or not digested at all in the in vivo situation, but rather that the proteins are digested more slowly. The protein compositions according to the invention thus allow the formulation of infant milk formulas that exhibit a digestibility profile that more closely resembles that of human breast milk (i.e., lower than the digestibility of standard infant milk formula). With human breast milk being the gold standard, the protein compositions thus produce improved protein digestibility.Thus, "improved" protein digestibility refers to lower / less protein digestibility (for example, since breast milk has lower / less digestibility than the reference formula in Figure 1).

[013] Consequently, in a first aspect, the invention relates to a protein composition for an infant milk formula, wherein the protein composition comprises casein, whey protein and at least one non-dairy protein with a high isoelectric point (i.e., a non-dairy source protein with a high isoelectric point) having an isoelectric point between 6.0 and 11, wherein the ratio, by weight, of the sum of casein and whey protein to the milk protein with a high isoelectric point is between 95:5 and 75:25.

[014] According to a second aspect, the invention relates to a protein composition suitable for an infant milk formula, wherein the composition comprises casein and whey protein in a weight ratio between 40:60 and 55:45, the composition comprises 10 to 40% by weight, based on the total protein content, of at least one protein with a high isoelectric point having an isoelectric point between 6.0 and 11. Petition 870260056360, dated 10 / 06 / 2026, p. 23 / 84 5 / 29

[015] In a third aspect, the invention relates to an infant milk formula comprising the protein composition of the present invention. The infant milk formula is suitable for administration to infants aged between 0 and 36 months.

[016] Without adhering to theory, it is hypothesized that electrostatic interactions and / or the protein structure of proteins with a high isoelectric point advantageously reduce the digestibility of the entire protein composition. Lower digestibility, as in breastfed infants, advantageously translates into optimal morphological and functional development of the infant's intestinal sections, reduced amino acid oxidation, promotion of amino acid deposition, and improved tissue development.

[017] Consequently, the present invention also relates to an infant formula to improve nutrient absorption, to promote amino acid deposition and to promote tissue development in infants, particularly in infants with a need for them, such as premature infants. Clauses 1. A protein composition suitable for an infant milk formula, wherein the composition comprises casein, whey protein and at least one protein with a high isoelectric point from a non-dairy source, wherein said protein with a high isoelectric point has an isoelectric point between 6.0 and 11, in which the ratio, by weight, of the sum of casein and whey protein to the protein with a high isoelectric point in the composition is between 95:5 and 75:25. 2. The protein composition according to clause 1, wherein casein and whey protein are present in a ratio, by weight, between 30:70 and 70:30, preferably between 38:62 and 62:38, more preferably around 50:50 or around 40:60. 3. The protein composition, according to clause 1 or 2, wherein the casein comprises β-caseins in an amount between 35 and 60%, by weight, of the total casein content. Petition 870260056360, dated 10 / 06 / 2026, page 24 / 84 6 / 29 4. The protein composition according to any previous clause, wherein casein comprises α-caseins in an amount between 20 and 60%, by weight, of the total casein content. 5. The protein composition according to any previous clause, wherein casein comprises k-caseins in an amount between 15 and 30%, by weight, of the total casein content. 6. The protein composition according to any previous clause, wherein the whey protein comprises α-lactalbumin in an amount between 20 and 80%, by weight, of the total whey protein content. 7. The protein composition according to any previous clause, wherein the whey protein comprises β-lactoglobulin in an amount between 20 and 70%, by weight, of the total whey protein content. 8. The protein composition according to any preceding clause, wherein the protein with a high isoelectric point is one or more proteins selected from the group consisting of lysozyme, ovotransferrin, canola protein, rapeseed protein, or mixtures thereof. 9. A protein composition suitable for an infant milk formula, wherein the composition comprises casein and whey protein in a weight ratio between 40:60 and 55:45, the composition comprises 10 to 40% by weight, based on the total protein content, of at least one protein with a high isoelectric point that has an isoelectric point between 6.0 and 11. 10. The protein composition, according to clause 9, in which at least one protein with a high isoelectric point is present in an amount of 15 to 35% by weight, preferably 18 to 30% by weight, based on the total protein content. 11. The protein composition, according to clause 9 or 10, in which at least one protein with a high isoelectric point is selected from lactoferrin, lysozyme, ovotransferrin, canola protein, rapeseed protein, or mixtures thereof, preferably lactoferrin, lysozyme, ovotransferrin, or mixtures thereof, with lactoferrin being the most preferred. Petition 870260056360, dated 10 / 06 / 2026, p. 25 / 84 7 / 29 12. The protein composition conforms to any of the preceding clauses, wherein at least one protein with a high isoelectric point is in intact form. 13. The protein composition is in accordance with any previous clause, in which only one protein with a high isoelectric point that has an isoelectric point between 9.0 and 11 is present. 14. The protein composition according to any of clauses 1 to 12, wherein two proteins with high isoelectric points that have an isoelectric point between 6.0 and 11 are present, wherein one of the isoelectric proteins has an isoelectric point between 9.0 and 11. 15. The protein composition according to any of the preceding clauses, consisting of casein, whey protein and at least one protein with a high isoelectric point, preferably one or two proteins with a high isoelectric point, including a protein with a high isoelectric point from a non-dairy source. 16. An infant milk formula comprising the protein composition as defined in any of clauses 1 to 14, suitable for administration to an infant aged between 0 and 36 months. 17. The protein composition according to any of clauses 1 to 14, or the infant milk formula according to clause 15, for therapeutic use in improving digestibility, preferably protein digestibility, in an infant who suffers from or is at risk of suffering from impaired digestibility and / or immature tissue development and / or immature gastrointestinal development, preferably premature infants. 18. Use of the protein composition in accordance with any of clauses 1 to 14, or of the infant milk formula in accordance with clause 15, to improve protein digestibility in a healthy human infant, and / or to promote tissue development in a healthy human infant. 19. A (non-therapeutic) method for improving digestibility, preferably protein digestibility, in a human infant. Petition 870260056360, dated 10 / 06 / 2026, p. 26 / 84 8 / 29 healthy, in which a protein composition is administered to the infant in accordance with any of clauses 1 to 14, or an infant milk formula in accordance with clause 15. 20. Use of the protein composition in accordance with any of clauses 1 to 14, in the manufacture of an infant milk formula to improve protein digestibility in a human infant, and / or to promote tissue development in a human infant. Detailed Description of the Invention: Protein Compositions

[018] As used in the present application, “protein compositions” refers to compositions comprising only protein material (peptides, free amino acids, proteins), and preferably do not contain other components that contribute to the caloric content of the composition, such as carbohydrates, fats and fiber. Expressed differently, the term “protein composition” in the context of the invention does not encompass nutritional compositions comprising other nutrients such as fats, carbohydrates, etc.

[019] Typically, the protein composition according to the invention has a pH in the range of 6.5 to 7.5.

[020] The protein compositions according to the invention comprise casein and whey protein. Casein and whey protein preferably represent 65 to 95% by weight of the protein, more preferably 70 to 90% by weight, up to more preferably 75 to 85% by weight of the total protein weight. In a preferred embodiment, the remainder of the protein is formed from non-dairy protein with a high isoelectric point, as defined in the present application.

[021] In one embodiment, the ratio, by weight, of the sum of casein and whey protein to at least one protein with a high isoelectric point is preferably between 95:5 and 75:25, more preferably 90:10 to 75:25, up to most preferably 85:15 to 75:25. Petition 870260056360, dated 10 / 06 / 2026, page 27 / 84 9 / 29 Casein

[022] Typically, in the protein composition according to the invention, casein is present in a ratio, by weight, to whey proteins between 25:75 and 70:30, more preferably 30:70 and 70:30, even more preferably between 38:62 and 62:38, with the highest preference being about 50:50 or about 40:60. Even more preferably, it is a casein to whey protein ratio between 25:75 and 62:38, preferably 30:70 and 55:45, with the highest preference being 40:60 and 52:48.

[023] The casein used in the present invention is preferably, but not necessarily, bovine casein. Typically, the casein is used in the form of casein micelles, as obtained from skimmed milk powder. The casein micelles comprise alphaS1, alphaS2, beta and kappa-caseins, the ratios of which may differ according to the supplier. In a preferred embodiment, the casein does not include caseinates and / or dissociated casein micelles.

[024] In a preferred embodiment, the casein fraction is enriched in beta-caseins. Consequently, in one embodiment, the protein compositions comprise beta-casein in an amount of 35 to 60% by weight, preferably 38 to 55% by weight, more preferably 40 to 50% by weight, based on the total casein content. For comparison purposes, conventionally, beta-casein is present in casein in amounts of approximately 33% by weight.

[025] In a preferred embodiment, the casein fraction is enriched in kappa-casein. Consequently, in one embodiment, the protein compositions comprise a casein fraction comprising kappa-casein between 15 and 30%, by weight, of the total casein content, preferably between 15 and 28%, by weight, more preferably between 18 and 25%, by weight, of the total casein content. Preferably, kappa-casein does not represent more than 40%, by weight, of the sum of alpha, beta and kappa-caseins, more preferably not more than 35%, by weight, up to most preferably not more than 30%, by weight; but Petition 870260056360, dated 10 / 06 / 2026, page 28 / 84 10 / 29 not less than 10%, by weight, more preferably not less than 15%, by weight, up to more preferably not less than 20%, by weight, of the sum of alpha, beta and kappa caseins. For comparison purposes, kappa casein is conventionally present in casein in quantities of approximately 33%, by weight.

[026] In a preferred embodiment, the sum of beta and kappa-caseins in the casein fraction is between 50 and 80%, by weight, of the total casein content, preferably 60 to 75%, by weight, of the total casein content.

[027] Typically, the protein composition comprises alpha-caseins in an amount between 20 and 60% by weight, preferably 25 and 55% by weight, more preferably 30 and 50% by weight, of the total casein content. Preferably, the sum of aS1-casein and aS2-casein represents not less than 20% by weight, based on the sum of alpha, beta and kappa casein, preferably not less than 25% by weight; but not more than 40% by weight, based on the sum of alpha, beta and kappa casein, preferably not more than 35% by weight.

[028] In contrast to commercially available standard infant formulas, the infant formula of the invention may comprise excess beta-casein relative to alpha-casein, preferably alphaS2-casein. Consequently, in one embodiment, the alpha-caseins are present in a ratio to beta-casein of at least 55:45. Whey Protein

[029] “Whey protein”, also known in the art as “whey proteins”, as used in the present application, is intended to refer to the major proteins as well as the minor whey proteins. “Major whey proteins” include alpha-lactalbumin and beta-lactoglobulins, depending on the whey source. Preferably, the major whey proteins used in the present invention include both beta- and alpha-lactalbumin. “Minor whey proteins” are preferably selected from serum albumin, proteose peptone, Petition 870260056360, dated 10 / 06 / 2026, p. 29 / 84 11 / 29 immunoglobulins, lactoferrin and lysozyme. In the context of the invention, the terms “whey” and “whey protein” are used interchangeably, and refer to the protein component of whey.

[030] Typically, whey proteins that may be included in the protein compositions of the invention include lactalbumins, beta-lactoglobulin, serum albumin, proteose peptone, immunoglobulins, lactoferrin, etc., all according to their conventional meaning in the field.

[031] Preferably, the whey protein of the invention's composition is enriched with alpha-lactalbumin. Typically, α-lactalbumin is present in the protein composition in an amount between 20 and 80% by weight of the total whey content, more preferably 30 and 80% by weight, up to more preferably 40 and 75% by weight, with the highest preference being 45 and 70% by weight. Alpha-lactalbumin is suitably obtained as purified alpha-lactalbumin ingredients or whey protein concentrates enriched with alpha-lactalbumin. For comparison purposes, conventionally, whey protein comprises about 27% by weight of alpha-lactalbumin.

[032] Whey protein also comprises β-lactoglobulin. Preferably, beta-lactoglobulin is present in an amount between 20 and 70%, by weight, of the total whey content, preferably 25 and 60%, by weight, up to more preferably 30 and 55%, by weight.

[033] Preferably, the ratio of α-lactalbumin to β-lactoglobulin is 1:1 to 6:1, more preferably 2:1 to 5:1. In particular, it is believed that better results are achieved in reducing digestibility, particularly of proteins, when the protein fraction of the invention comprises α-lactalbumin in an amount of at least 20%, by weight, of the total whey protein content. In a more preferred embodiment, better results in reducing digestibility, particularly of proteins, have been observed when the protein compositions of the invention comprising α-lactalbumin in an amount of Petition 870260056360, dated 10 / 06 / 2026, p. 30 / 84 12 / 29 at least 20%, by weight, of the total whey protein content are combined with a casein fraction that is enriched in β-casein, that is, in an amount of at least 28%, by weight, of the total casein content.

[034] Some secondary whey proteins, for example, immunoglobulins and lactoferrin, may have an isoelectric point of at least 6.0. In one embodiment, the whey comprises a milk protein with an isoelectric point between 6.0 and 11 in an amount of less than 0.2% by weight, preferably less than 0.1% by weight, up to more preferably less than 0.05% by weight, based on the total protein content. In a more preferred embodiment, said protein is lactoferrin. In a still more preferred embodiment, the infant formula comprises lactoferrin in an amount of less than 0.2% by weight, preferably less than 0.1% by weight, up to more preferably less than 0.05% by weight, based on the total protein content.That is, in one embodiment, the infant formula of the invention, non-dairy proteins with a high isoelectric point replace secondary whey proteins, particularly secondary whey proteins selected from proteose peptone, immunoglobulins, and lactoferrin.

[035] In one embodiment, serum albumin is present in an amount of not more than 2% by weight, based on the total whey content, preferably not more than 1% by weight, more preferably not more than 0.1% by weight. Protein(s) with High Isoelectric Point (pI)

[036] The protein compositions according to the invention comprise at least one protein with a high isoelectric point.

[037] As used in the present application, a “high isoelectric point protein” is defined as a protein with an isoelectric point of 6.0 to 11, preferably at least 6.5, more preferably at least 7.0, up to most preferably at least 7.5. The high isoelectric point protein in the context of the invention is selected from a single high isoelectric point protein or a mixture of at least two high isoelectric point proteins. Petition 870260056360, dated 10 / 06 / 2026, page 31 / 84 13 / 29 high isoelectric point (that is, each of the proteins with a high isoelectric point has an isoelectric point of 6.0 to 11, preferably in the range of 7.0 to 10).

[038] In one embodiment, at least one protein with a high isoelectric point is of non-dairy origin. As used in the present application, “non-dairy” or “non-milk-based” protein has the usual meaning that the protein was not taken from or extracted from the mammary gland of a mammal.

[039] Consequently, in one aspect, the invention relates to a protein composition for an infant milk formula, wherein the protein composition comprises casein, whey protein and at least one non-dairy protein with a high isoelectric point having an isoelectric point between 6.0 and 11, wherein the ratio, by weight, of the sum of casein and whey protein to the high isoelectric point milk protein is between 95:5 and 75:25.

[040] Suitable examples of non-dairy proteins with a high isoelectric point having a pI between 6.0 and 11 for use in the present invention include, but are not limited to, proteins extracted from eggs (e.g., lysozyme), peas, soy, canola, rapeseed, or combinations thereof.

[041] Preferably, at least one non-dairy protein with a high isoelectric point according to the invention is selected from ovotransferrin, lysozyme, rapeseed or combinations thereof; more preferably ovotransferrin and lysozyme.

[042] In another embodiment, at least one high isoelectric point protein is a high isoelectric point protein of dairy origin, i.e., from a mammal. Suitable examples of high isoelectric point dairy proteins having a pI between 6.0 and 11 for use in the present invention include, for example, immunoglobulins (pI of 5.5 to 8.3), lactoferrin (pI of about 8.9) and lysozyme (pI of about 10.4). Preferably, at least one high isoelectric point protein from a dairy source is selected from lactoferrin and lysozyme, more preferably lactoferrin.

[043] Consequently, in another aspect, the invention relates to a Petition 870260056360, dated 10 / 06 / 2026, p. 32 / 84 14 / 29 Protein composition for an infant milk formula, wherein the protein composition comprises casein and whey protein in a weight ratio between 40:60 and 55:45, the composition comprises 10 to 40% by weight, based on the total protein content, of at least one high isoelectric point protein having an isoelectric point between 6.0 and 11. The high isoelectric point protein may be a high isoelectric point milk protein, either alone or in combination with a high isoelectric point non-milk protein.

[044] In yet another embodiment, the protein composition comprises a mixture of dairy and non-dairy proteins with a high isoelectric point having a pI between 6.0 and 11. In this case, the dairy product is preferably lactoferrin and the non-dairy product is preferably selected from ovotransferrin, lysozyme and / or rapeseed.

[045] At least one protein with a high pI for use according to the invention is preferably selected from lactoferrin, lysozyme, ovotransferrin, canola protein, rapeseed protein or mixtures thereof, preferably lactoferrin, lysozyme, ovotransferrin or mixtures thereof, with lactoferrin being the most preferred.

[046] Preferably, at least one protein with a high isoelectric point is present in the protein compositions in an amount of 15 to 35% by weight, preferably 18 to 30% by weight, based on the total protein content.

[047] According to a preferred embodiment, the protein compositions according to the invention comprise a single high isoelectric point protein having an isoelectric point between 9.0 and 11. When used individually, the high isoelectric point protein has an isoelectric point of at least 9.5, more preferably at least 10.0. Preferably, the high isoelectric point protein is lysozyme, more preferably egg lysozyme. Suitable amounts of egg lysozyme, when used as the sole high isoelectric point protein, are in the range of 15 to 30% by weight based on the total protein content, more preferably 18 to 23% by weight. Petition 870260056360, dated 10 / 06 / 2026, page 33 / 84 15 / 29 weight.

[048] According to another preferred embodiment, the protein compositions according to the invention comprise several high pI proteins, such as mixtures of high pI dairy and non-dairy proteins in the amounts prescribed according to the invention, i.e., added in excess to high pI proteins, possibly in the whey fraction. In a preferred embodiment, more than two high pI proteins having an isoelectric point between 6.0 and 11 are present, wherein at least 50% of the proteins have an isoelectric point between 9.0 and 11. Even more preferably, two high pI proteins having an isoelectric point between 6.0 and 11 are present in the protein composition. The preferred combination of two high pI proteins is ovotransferrin and lysozyme.

[049] Properly, the two or more high pI proteins are present in a ratio wherein the protein with the higher pI is present in an amount at least 50% greater than the high pI protein that has the lower pI. Put differently, in a protein composition comprising two or more high pI proteins that have a pI between 6.0 and 11, the high pI protein that has the lower pI preferably does not represent more than 50% of the total high pI protein content, more preferably not more than 30%, by weight, up to more preferably not more than 20%.

[050] Typically, the protein composition of the invention comprises at least one protein with a high isoelectric point in an amount of 5 to 35% by weight, based on the total weight of proteins, more preferably from 10 to 30% by weight, up to more preferably from 15 to 25% by weight of the total weight of proteins.

[051] In one embodiment, the content of high isoelectric point protein replaces some of the whey proteins, preferably secondary whey proteins, more preferably secondary whey proteins selected from proteose peptone, immunoglobulins and / or lactoferrin. Consequently, at least one protein with a high isoelectric point Petition 870260056360, dated 10 / 06 / 2026, page 34 / 84 16 / 29 high as a “secondary whey protein” is present in an amount of 5 to 30% by weight, up to more preferably between 10 and 25% by weight, with the most preference of 15 to 20% by weight, based on the total whey protein content. Preferably, the protein compositions of the invention comprise proteins with a high isoelectric point, as a substitute for “secondary whey protein” in an amount of 2.5 to 15% by weight, preferably 5 to 10% by weight, based on the total protein weight.

[052] Proteins with a high isoelectric point may be hydrolyzed, partially hydrolyzed, or intact. Preferably, proteins with a high isoelectric point are in intact form. The term “an intact protein,” as used in this application, refers to any form of intact protein, including but not limited to, a protein concentrate and / or a protein isolate, as well as other forms of intact proteins.

[053] Based on the final weight of infant milk formulas comprising any of the protein compositions according to the invention, the formula comprises from 5 to 25%, by weight, more preferably at least 10 to 20%, by weight, based on the dry weight of the formula, of a protein having an isoelectric point between 6.0 and 11.

[054] Preferably, after reconstitution, the protein having an isoelectric point between 6 and 11 is present in an amount between 0.01 and 0.33 g / 100 mL, more preferably 0.1 to 0.3 g / mL. Infant Milk Formula

[055] In one aspect of the invention, the protein compositions described in the present application are included in an infant formula product.

[056] Consequently, in one embodiment, the invention relates to an infant milk formula comprising a protein composition according to claim 1, that is, a protein composition for an infant milk formula, wherein said protein composition comprises casein, whey protein and at least one non-dairy protein with a high isoelectric point that has Petition 870260056360, dated 10 / 06 / 2026, page 35 / 84 17 / 29 an isoelectric point between 6.0 and 11, where the ratio, by weight, of the sum of casein and whey protein to milk protein with a high isoelectric point is between 95:5 and 75:25.

[057] In another embodiment, the invention relates to an infant milk formula comprising the protein composition according to claim 9, that is, a protein composition for an infant milk formula, wherein the protein composition comprises casein and whey protein in a ratio, by weight, between 40:60 and 55:45, the composition comprising 10 to 40%, by weight, based on the total protein content, of at least one protein with a high isoelectric point having an isoelectric point between 6.0 and 11.

[058] All limitations for protein compositions as described above apply with the necessary modifications (mutatis mutandis).

[059] The term “infant formula” is well defined and consistently controlled internationally by regulatory bodies. In particular, CODEX STAN 73 - 1981 “Standard For Infant Formula and Formulas For Special Medical Purposes Intended for Infants” is widely accepted. It recommends the nutritional value and composition of the formula, requiring that the prepared milk contain no less than 60 kcal (250 kJ) and no more than 70 kcal (295 kJ) of energy per 100 mL. The FDA and other regulatory bodies have established nutritional requirements in accordance with the same. The terms “infant formula” and “infant milk formula” used in this application are interchangeable.

[060] The infant formula according to the invention can be used as a nutritional composition, nutritional therapy, nutritional support, as a medical food, as a food for special medical purposes, or as a nutritional supplement. The present infant formula is preferably an enteral (oral) composition. The infant formula is administered orally, or is intended to be administered orally, to a subject in need thereof, in particular to children and infants, including children who are Petition 870260056360, dated 10 / 06 / 2026, p. 36 / 84 18 / 29 starting to walk, preferably children up to 6 years of age, preferably infants or toddlers, typically aged 0 to 36 months, more preferably infants aged 0 to 12 months, with the highest preference for 0 to 6 months of age. Thus, in some embodiments, the present infant formula is an infant formula for infants aged 0 to 6 months, or a follow-on formula for infants aged 6 to 12 months, or a toddler formula (also called “growing-up milk”) for children who are starting to walk aged between 12 and 36 months.

[061] In order to meet the caloric requirements of the infant, the present infant formula preferably comprises 50 to 200 kcal / 100 mL of liquid, more preferably 60 to 90 kcal / 100 mL of liquid, up to most preferably 60 to 75 kcal / 100 mL of liquid. This caloric density ensures an optimal ratio between water and calorie intake. The osmolarity of the present infant formula is preferably between 150 and 420 mOsmol / L, more preferably 260 to 320 mOsmol / L. The low osmolarity aims to reduce gastrointestinal stress.

[062] Preferably, the present infant formula is in liquid form, preferably with a viscosity below 35 mPa.s, more preferably below 10 mPa.s, even more preferably below 6 mPa.s, as measured in a Brookfield viscometer at 20 °C, at a shear rate of 100 s-1. Suitably, the present infant formula is in powder form, which preferably can be reconstituted with water to form a liquid, or in concentrated liquid form, which must be diluted with water. When the present infant formula is in liquid form, the preferred volume administered daily is in the range of about 80 to 2500 mL, more preferably about 450 to 1000 mL per day.

[063] The infant formula of the invention is heat-treated. The term “heat treatment” in this application refers to treatment at an elevated temperature, aimed at increasing the shelf life of the infant formula. Examples Petition 870260056360, dated 10 / 06 / 2026, page 37 / 84 19 / 29 heat treatment methods include pasteurization, sterilization, and UHT (ultra-high temperature treatment or ultra-high temperature processing).

[064] Appropriately, the invention formula is not or does not comprise mammalian milk, in particular, the invention formula is not or does not comprise human breast milk. Other Components

[065] The infant formula according to the invention may suitably comprise typically infant formula components in usual amounts. The infant formula may thus comprise at least one of the following proteins (in addition to casein, whey and the protein(s) with a high isoelectric point, as described in the present application), fatty acids (e.g., long-chain polyunsaturated fatty acids), animal fat, vegetable oils or mixtures thereof as a fat fraction, digestible carbohydrate fraction (e.g., lactose), indigestible carbohydrate fractions and may comprise dietary fiber with prebiotic activity, probiotic bacteria, a vitamin-mineral mixture, etc.

[066] The infant formula according to the invention preferably comprises a lipid component, preferably a lipid component suitable for infant nutrition as known in the art. The lipid component of the present infant formula preferably provides 2.9 to 6.0 g, more preferably 4 to 6 g per 100 kcal of the composition. When in liquid form, the infant formula typically 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 infant formula preferably comprises 12.5 to 40% by weight of lipid, more preferably 19 to 30% by weight.

[067] The infant formula according to the invention may further comprise proteinaceous material, in addition to casein, whey and protein with a high isoelectric point, according to the invention. In the context of the present invention, the additional “protein”, “proteinaceous material” or “protein equivalents” includes proteins, peptides, free amino acids and proteins. Petition 870260056360, dated 10 / 06 / 2026, page 38 / 84 20 / 29 partially or extensively hydrolyzed. Preferably, the additional proteinaceous material, besides beta-lactoglobulin derivative(s), does not cause an allergic reaction, or is hypoallergenic, such as free amino acids and hydrolyzed protein.

[068] As an additional protein component, the infant formula according to the present invention preferably comprises free amino acids, hydrolyzed whey protein, preferably partially hydrolyzed whey proteins.

[069] The present infant formula preferably contains at least 50% by weight of milk-derived (non-human) protein component, more preferably at least 90% by weight, based on the dry weight of the total protein content.

[070] The present infant formula preferably contains 5 to 20%, more preferably 7 to 16%, with a maximum preference of 7 to 12% protein, based on total calories. The present infant formula, when in liquid form, preferably contains 0.5 to 6.0 g, more preferably 0.8 to 3.0 g, up to most preferably 1.0 to 2.5 g of protein per 100 mL. The present infant formula preferably comprises at least 7%, by weight, more preferably at least 8%, by weight, with a maximum preference of at least 9% or at least 10% protein based on the dry weight of the total infant formula. Preferably, the present infant formula comprises at least 40%, by weight, more preferably a maximum of 15%, by weight, preferably a maximum of 20%, by weight, of the protein composition, based on the dry weight of the total infant formula.

[071] Infant formula may comprise digestible carbohydrate(s). Typically, digestible carbohydrates are used that are known in the art to be suitable for use in infant nutritional compositions, for example, selected from digestible polysaccharides (e.g., starch, maltodextrin), digestible monosaccharides (e.g., glucose, fructose), digestible disaccharides (e.g., lactose, sucrose). It Petition 870260056360, dated 10 / 06 / 2026, page 39 / 84 21 / 29 particularly suitable lactose and / or maltodextrin.

[072] The digestible carbohydrate component preferably comprises at least 60% lactose by weight, based on total digestible carbohydrate, more preferably at least 75% by weight, up to most preferably at least 90% lactose, based on total digestible carbohydrate. Human Subjects

[073] The formula of any aspect of the present invention is intended for a human infant. Suitably, the formula of any aspect of the present invention is intended for a human term infant and / or human preterm infant. Suitably, the present infant formula is to provide the daily nutritional requirements for a human infant, in particular, for administration, in particular, for feeding humans, in particular, infants including toddlers. The human subjects or target population are preferably infants (0 to 12 months) and toddlers (up to 36 months), preferably infants and toddlers who are partially or exclusively fed non-human milk. In the context of the invention, a “human term infant” is an infant born after 37 weeks of gestation, preferably between 37 and 42 weeks of gestation.Consequently, a "human preterm infant" is an infant born before completing 37 weeks of gestation.

[074] In one embodiment, the human infant or child who is beginning to walk is fed infant formula as well as human milk. In another embodiment, the human infant or child is exclusively fed infant formula. Digestibility

[075] In an ideal world, protein digestibility, in particular casein and whey protein digestibility, in formula-fed infants is similar to that observed in breastfed infants. Petition 870260056360, dated 10 / 06 / 2026, p. 40 / 84 22 / 29 maternal. However, the digestibility profile of human milk is considerably lower than the digestibility of standard commercial infant formula. Figure 1 shows a comparison between the total appearance of NH2 over time under intestinal digestion conditions of fresh human milk (represented as squares; used on the same day it was collected) and a commercial infant formula (represented as triangles, commercial infant milk formula (IMF)), prepared according to package instructions and reconstituted at 1.3% w / w protein.

[076] As used in this application, “low” / “lower” protein digestibility does not imply that proteins are digested to a lesser degree or not digested at all in the in vivo situation, but rather that proteins are digested more slowly. In this application, “protein digestibility” refers to the total protein digested in the “in vitro” static digestion model for infants over 3 h of digestion.

[077] The low digestibility of human milk is beneficial to ensure adequate nutrient absorption in the intestinal phase, in particular, the proper breakdown of proteins and peptides, and the absorption of amino acids for tissue development. A faster protein digestibility, as in standard infant formula, results in the postprandial appearance of amino acids in the blood, stimulating amino acid oxidation and ultimately adversely affecting tissue development in infants.

[078] It is also believed that the lower digestibility of human milk in infants / children who are beginning to walk promotes the adequate supply of proteins, peptides and amino acids in specific sections of the gastrointestinal tract, which are responsible for signaling the morphological and functional development of the intestinal section referred to.

[079] Infants not fed breast milk will be more susceptible to the consequences of impaired or suboptimal intestinal digestion of protein compositions. These consequences may be observed during early childhood (i.e., a direct effect) or may be Petition 870260056360, dated 10 / 06 / 2026, page 41 / 84 23 / 29 observed later in life. Furthermore, the consequences of suboptimal or deficient development of intestinal function or nutrient absorption in formula-fed infants are not only immediate but can also persist into adulthood, creating a basis for increased susceptibility to a range of diseases and conditions, such as obesity, reduced immunity, etc.

[080] Abnormal rapid digestibility affects nutrient absorption, which can ultimately lead to delayed tissue development and growth in infants, and impaired development of intestinal function. Impaired digestibility can be measured in vitro as a percentage of digestion hydrolysis over time or preferably at a physiologically relevant point in time.

[081] Advantageously, the protein compositions according to the invention have a lower protein digestibility profile compared to compositions used in standard commercial infant formulas.

[082] Impaired protein digestibility can lead to diseases and conditions related to the growth and development of infants, in particular, tissue development. This is particularly relevant in low birth weight infants or infants who suffer from or are at risk of suffering from diseases or conditions that affect intestinal nutrient absorption. In these cases, it is particularly desirable that protein compositions be digested slowly in order to ensure optimal nutrient absorption and amino acid influx into the bloodstream. Impaired or suboptimal protein digestibility can be avoided or treated by using the protein compositions according to the invention.Consequently, in one aspect, the invention relates to the use of the protein compositions or infant milk formula described in this application for therapeutic use in improving digestibility, preferably protein digestibility, in an infant who suffers from or is at risk of suffering from impaired digestibility and / or delayed tissue development and / or delayed gastrointestinal development. Petition 870260056360, dated 10 / 06 / 2026, p. 42 / 84 24 / 29 Premature infants can especially benefit since their gastrointestinal tract is characterized by being immature.

[083] Associated with these findings above, the invention relates to the protein composition and infant formula as defined in the present application for therapeutic use in improving digestibility, preferably protein digestibility, in an infant who suffers from or is at risk of suffering from impaired digestibility and / or delayed tissue development and / or delayed gastrointestinal development, or premature infants with immature tissue development, in particular, of the gastrointestinal tract. Worded differently, the invention relates to the use of the protein composition and infant formula as defined in the present application to improve protein digestibility in a healthy human infant, and / or to promote tissue development in a healthy human infant.The invention also relates to a (non-therapeutic) method for improving digestibility, preferably protein digestibility, in a healthy human infant, wherein a protein composition or infant formula is administered to the infant as defined in this application. Worded differently, the invention relates to the use of the protein composition as defined in this application in the manufacture of an infant milk formula to improve protein digestibility in a human infant, and / or to promote tissue development in a human infant. In the context of the invention, the term "improving digestibility" means reducing digestibility relative to the digestibility obtained for conventional infant formula, i.e., reducing the digestibility gap compared to the digestibility profile obtained for human milk.In one embodiment, this conventional infant formula has a protein composition produced from casein and whey protein with a casein:whey protein ratio, by weight, of approximately 40:60 (as demonstrated in Example 1). Associated with this, its use extends to promoting tissue development in a healthy human infant and improving the absorption of protein, peptides, and / or... Petition 870260056360, dated 10 / 06 / 2026, page 43 / 84 25 / 29 to amino acid absorption in a healthy human infant.

[084] The infant or child who is beginning to walk may suffer from, or is at risk of suffering from, growth retardation or delayed tissue development. Preferably, the infant or child who is beginning to walk is a preterm human infant or a term human infant, preferably a preterm or underweight human infant. Examples Example 1 Protein Composition and Digestibility

[085] The protein compositions, as described in Table 1, were prepared as follows:

[086] Whey proteins and / or proteins with high pI were dissolved in demineralized water at room temperature under gentle stirring for 2 hours. Caseins were dispersed in demineralized water at 50 °C and homogenized at 350 ± 50 bar. The final protein formulations were obtained by mixing dissolved protein ingredients in appropriate proportions to achieve a final protein concentration of 1.3% (w / w). The amount of proteins with high pI is expressed as a percentage, by weight, based on the total protein content. Table 1 Protein Composition Components Ratio, by weight, of casein:whey protein Ref. IMF Standard casein 1 Standard whey protein 2 40:60 Ref. P Standard casein 3 Standard whey protein 2 50:50 Ref. P-beta casein enriched with beta-casein 5 Standard whey protein 2 50:50 A Standard casein 3 enriched whey protein 50:50 Petition 870260056360, dated 10 / 06 / 2026, page 44 / 84 26 / 29 Protein Composition Components Ratio, by weight, of casein: whey protein with alpha-lactalbumin4 ovotransferrin 15%, by weight6 lysozyme 3%, by weight7 B casein enriched with beta-casein5 whey protein enriched with alpha-lactalbumin4 ovotransferrin 15%, by weight6 lysozyme 3%, by weight7 50:50 1: skimmed milk 2: whey protein concentrate powder (Omia), demineralized 80% (Friesland) Campina)3: micellar casein isolate (Friesland Campina)4: Lacprodan (Arla; alpha-lactalbumin content 60%) 5: Ultranor HT (Kerry Foods; beta-casein content 40%) 6: pI = 6.7, non-dairy source (Bioseutica Group) 7: pI = 10.4, non-dairy source (Bioseutica Group)

[087] The protein compositions in Table 1 were digested using the INFOGEST static in vitro digestion model adapted for infants, as shown in Table 2 below: Table 2 Gastric Phase (60 min) Intestinal Phase (120 min) Meal Ratio: Simulated Gastric Fluid 63:37 Meal Ratio: Simulated Gastric Fluid 39:61 Enzymes: Pepsin (268 U / mL) and Lipase (19 U / mL) from rabbit gastric extract Enzymes: Porcine pancreatin (90 U / mL) and bovine bile (3.1 mM) pH 4.5, T = 37 °C pH 6.6, T = 37 °C

[088] Digestions of 100 mL volumes were performed in bottles Petition 870260056360, dated 10 / 06 / 2026, page 45 / 84 27 / 29 A 250 mL Duran tube was used. A water bath (MixBathPC, 2mag Magnetic Notion) and a magnetic stirrer (Mixcontrol 40, 2mag Magnetic Notion) were set to 37.5 °C and 100 rpm, respectively. For the gastric phase, the protein solution set at 37 °C and simulated gastric fluid (SGF) were mixed in a 63:37 ratio. The pH was adjusted to 4.5 using 0.25M HCl, and the digested gastric contents were sampled at 5, 15, 30, and 60 minutes. After the gastric phase, the digested gastric contents were mixed with simulated intestinal fluid (SIF) in a 39:61 ratio. The pH was adjusted to 6.6 using 0.25 M NaOH / 0.25 M NaHCO3. Digested intestinal contents were sampled at 2, 6, 10, 15, 22, 30, 40, 55, 70, 80, 90, 105, and 120 minutes.

[089] Sampling was performed by mixing 1 mL of digested content with 1 mL of 0.1 M phosphate buffer at pH 7.2 (gastric samples) or 5.5 (intestinal samples) to stop digestion, dispersing (T10 Basic Ultraturrax, IKA) for 25 seconds at power 5.5 to increase protein dispersibility, flash-frozen in liquid nitrogen and stored at -20 °C until analysis. Each digestion was performed in triplicate.

[090] The collected samples were subjected to orthophthaldialdehyde (OPA) primary amino group generation analysis. The method is based on the specific reaction between OPA and free primary amino groups in proteins in the presence of 2-(dimethylamino)ethanethiol hydrochloride (DMA) to result in alkyl-iso-indole derivatives that show absorbance at 340 nm. Digested content samples were mixed with 2% SDS in a 1:1 ratio, boiled at 95 °C for 5 minutes, and allowed to cool to room temperature. 5 pL aliquots were mixed with 200 pL of OPA working reagent or OPA blank solution and incubated for 25 minutes. The signal was read using a Biowave plate reader, and the results were analyzed using Gen5 2.09 software. Free NH2 groups were quantified using the L-alanine solution calibration curve (0, 1, 2, 4, 6, 8 and 10 mM). Measurements were performed in duplicate. Petition 870260056360, dated 10 / 06 / 2026, page 46 / 84 28 / 29

[091] The results were expressed for the degree of hydrolysis (%) as a function of time. The digestibility parameter was obtained by fitting the data to the equation as described previously (Deng, van der Veer et al. 2017), and is shown in Figure 2.

[092] Ref. FMI represents the protein digestibility profile of standard infant formula. Ref. P and Ref. P-beta are used as a reference for the protein digestibility profile of formulations with casein and whey (50:50) without proteins with a high pI. Both Ref. P and Ref. P-beta have lower digestibility than Ref. FMI, regardless of the casein source.

[093] Improved (lower) digestibility is observed in compositions according to the invention (A, B), i.e., comprising proteins with a high isoelectric point, with even better results when the casein source is enriched in beta-casein. Example 2 Comparison of Protein with High pI

[094] Composition B of Table 1 (18%, by weight, based on total protein content, of proteins having a pI of at least 6.0; a 50:50 ratio of casein:whey protein; and the use of a beta-casein enriched casein source) was reproduced using different sources and ratios of high isoelectric point proteins. The compositions were prepared in the same manner as described in Example 1, and also had a final protein concentration of 1.3% (w / w). Table 3 Protein composition Components (% by weight, based on total protein content) C 1 Ovotransferrin 18% by weight D 2 Lysozyme 18% by weight E 3 Rapeseed protein 18% by weight F 4 Lysozyme 18% by weight Petition 870260056360, dated 10 / 06 / 2026, page 47 / 84 29 / 29 Protein composition Components (% by weight, based on total protein content) G ovotransferrin 9% by weight lysozyme 9% by weight 1: pl = 6.7 (Bioseutica Group) 2: pI = 8.7 (FrieslandCampina) 3: pI = 7 to 11 (Synthite Corporate) 4: pI = 10.4 (Bioseutica Group)

[095] The results are presented in Figure 3 (effect of single proteins with high pI) and Figure 4 (effect of two proteins with high pI). Figure 3 shows that the higher the pI of the protein used, the lower the protein digestibility. Figure 4 shows that a composition with a 50:50 ratio of two proteins with high pI has a synergistic effect in reducing protein digestibility. Petition 870260056360, dated 10 / 06 / 2026, page 48 / 84

Claims

1 / 3 Claims 1. INFANT MILK FORMULA comprising a protein composition, characterized by comprising casein and whey protein and at least one high isoelectric point protein having an isoelectric point between 6.0 and 11, wherein casein and whey protein are present in a weight ratio between 40:60 and 55:45, and the composition comprises 10 to 40% by weight, based on the total protein content, of at least one high isoelectric point protein having an isoelectric point between 6.0 and 11, wherein the at least one high isoelectric point protein is lactoferrin, wherein the high isoelectric point protein is in intact form, wherein the formula is suitable for administration to an infant aged between 0 and 36 months.

2. FORMULA, according to claim 1, characterized in that the casein comprises β-caseins in an amount between 35 and 60%, by weight, of the total casein content.

3. FORMULA, according to any one of claims 1 to 2, characterized in that the casein comprises α-caseins in an amount between 20 and 60%, by weight, of the total casein content.

4. FORMULA, according to any one of claims 1 to 3, characterized in that the casein comprises k-caseins in an amount between 15 and 30%, by weight, of the total casein content.

5. FORMULA, according to any one of claims 1 to 4, characterized in that the whey protein comprises α-lactalbumin in an amount between 20 and 80%, by weight, of the total whey protein content.

6. FORMULA, according to any one of claims 1 to 5, characterized in that the whey protein comprises β-lactoglobulin in an amount between 20 and 70%, by weight, of the total whey protein content.

7. FORMULA, according to any one of claims 1 to 6, characterized in that at least one protein with a high isoelectric point is present in an amount of 15 to 35% by weight, preferably 18 to 30% by weight, based on the total protein content.

8. FORMULA, according to any one of claims 1 to 7, characterized by a second protein with a high isoelectric point having an isoelectric point between 9.0 and 11 being present.

9. FORMULA, according to any one of claims 1 to 8, characterized in that the formula is suitable for administration to an infant aged between 0 and 12 months.

10. FORMULA, according to any one of claims 1 to 9, characterized by being for therapeutic use in reducing protein digestibility, compared to the digestibility obtained for conventional infant formula, in a human infant who suffers from or is at risk of suffering from impaired digestibility and / or immature tissue development and / or immature gastrointestinal development, preferably premature infants.

11. USE OF INFANT MILK FORMULA, as defined in any of claims 1 to 9, characterized by being for reducing protein digestibility, compared to the digestibility obtained for conventional infant formula, in a healthy human infant, and / or for promoting tissue development in a healthy human infant.

12. METHOD FOR REDUCING PROTEIN DIGESTIBILITY, compared to the protein digestibility obtained for conventional infant formula in a healthy human infant, characterized by administering to the infant an infant milk formula, as defined in any one of claims 1 to 9.

13. USE OF INFANT FORMULA, as defined in any of claims 1 to 9, characterized by being used in the manufacture of an infant formula product to reduce protein digestibility, compared to the protein digestibility obtained for conventional infant formula, in a human infant, and / or to promote tissue development in a human infant. (Petition 870260056360, dated 10 / 06 / 2026, p. 50 / 84 3 / 3)

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