Nutritional composition and infant formula for promoting brain myelination

Abstract

The present invention relates to nutritional compositions and infant formulas that promote brain myelination. More specifically, the present invention relates to nutritional compositions for infants and young children and related health benefits. The present invention relates to a nutritional composition for promoting or supporting an optimal myelination trajectory in the brain, in particular to a nutritional composition comprising a phospholipid, a metabolic precursor thereof and / or a metabolite thereof, for use in promoting or supporting an optimal myelination trajectory in the brain, such trajectory being close to the trajectory observed in infants fed only with human breast milk (HBM) several months before life.

Description

[0001] This application is a divisional application, and the parent application is an application with an application date of December 13, 2016, an application number of 201680072971.4, and an invention title of "Nutritional Compositions and Infant Formulas for Promoting Brain Myelination". Technical Field

[0002] The present invention relates to nutritional compositions for infants and young children and their associated health benefits. In particular, it relates to nutritional compositions comprising phospholipids, their metabolic precursors, and / or their metabolites, which are used to promote or support an optimal myelination trajectory in the brain, such a trajectory being close to that observed in infants fed solely with human breast milk (HBM) during the first few months of life. Background Art

[0003] Whenever a mother is unable to breastfeed her infant, infant formula can provide a suitable alternative to natural feeding with human breast milk. Nutritional compositions for infants and young children are typically sold as powders to be reconstituted with water or, in some cases, as ready-to-drink or concentrated liquid compositions. These compositions are designed to cover most or all of the nutritional needs of an infant or young child.

[0004] However, it is known that human breast milk represents the ultimate gold standard in terms of infant nutrition. Infant formula manufacturers have made many attempts to produce nutritional health effects that are close to or similar to those of human breast milk.

[0005] It is recommended that all infants be fed with their mother's milk. However, in some cases, due to certain medical reasons, breastfeeding is insufficient or impossible or unsuccessful, or the mother does not choose to breastfeed. Infant formulas have been developed for these situations.

[0006] However, in many instances, studies have shown that infant formulas do not have the same effects on the body as human breast milk.

[0007] For example, infants fed with infant formula and those fed with human breast milk (HBM) may exhibit different myelination trajectories. Myelination generally begins prenatally in the brain and continues postnatally until early adulthood.

[0008] Myelination is a fundamental process of neural development, which involves enclosing axons with fatty myelin sheaths that facilitate the transmission of action potentials. Myelination has been considered to play a key role in the coordinated communication between brain cells and networks. Myelinated white matter matures along with cognitive and learning abilities.

[0009] Infancy and early childhood, especially from the first few weeks after birth to 2 to 5 years of age, are sensitive periods for brain myelination, depending on the area of the brain.

[0010] A recent study (“Breastfeeding and early white matter development: a cross - sectional study”, Deoni et al., NeuroImage 82, (2013), 77 - 86) has demonstrated that breastfed children show increased white matter development in the late - maturing frontal regions and related brain areas compared to infants fed with infant formula. A positive correlation between white matter microstructure and breastfeeding duration has also been shown in several brain regions that are anatomically consistent with the observed improvements in cognitive and behavioral performance measures.

[0011] Thus, the results of this study suggest that not only is it important for myelination to occur to avoid cognitive deficits, but also that myelination occurs early in an infant's life and, ideally, follows the trajectory (myelination over time) that would occur in breastfed infants. In addition, the study shows that standard infant formulas generally do not provide this optimal myelination trajectory.

[0012] Accordingly, there is a need to provide a nutritional composition for infants or toddlers that will promote and / or support an optimal myelination trajectory in the brain, which is similar to the trajectory observed in infants fed only with human breast milk (HBM).

[0013] For infants fed with infant formula, there is a need to promote and / or support an optimal myelination trajectory in the brain, which is similar to the trajectory observed in infants fed only with human breast milk (HBM).

[0014] For infants fed with infant formula, there is also a need to provide them with optimal nutrition that can promote and / or support an optimal myelination trajectory in the brain, which is similar to the trajectory observed in infants breastfed only with human breast milk (HBM).

[0015] For infants fed with infant formula, there is a need to promote and / or support an optimal short - term or long - term health status through nutrition that promotes and / or supports an optimal myelination trajectory in the brain, which is similar to the trajectory observed in infants fed only with human breast milk (HBM); such health status includes optimal brain and cognitive function development and prevention of neurocognitive deficits.

[0016] There is a need to compensate for sub - optimal myelination observed in non - breastfed infants. SUMMARY OF THE INVENTION

[0017] The present invention relates to nutritional compositions for infants and young children, such as preterm infant formula, infant formula, follow-on formula, growing-up milk or baby foods, preferably infant formula. The composition comprises phospholipids, their metabolic precursors and / or their metabolites. The phospholipids, their metabolic precursors and / or their metabolites can promote or support an optimal myelination trajectory in the brain, which is close to the trajectory observed in infants fed only with human breast milk (HBM).

[0018] In one aspect, there is provided a nutritional composition for infants and / or young children, which comprises phospholipids, their metabolic precursors and / or their metabolites for promoting and / or supporting an optimal myelination trajectory in the brain, which is close to the trajectory observed in infants fed only with human breast milk (HBM).

[0019] In one aspect, there is provided a nutritional composition for infants and / or young children, which comprises phospholipids, their metabolic precursors and / or their metabolites for promoting and / or supporting an optimal myelination trajectory in the brain, which is close to the trajectory observed in infants fed only with human breast milk (HBM), and which results in promoting and / or supporting a healthy state characterized by optimal brain and cognitive function development and / or preventing neurocognitive deficits.

[0020] In another aspect, there is provided the use of phospholipids, their metabolic precursors and / or their metabolites for manufacturing a nutritional composition to be administered to infants and / or young children to promote and / or support an optimal myelination trajectory in the brain, which is close to the trajectory observed in infants fed only with human breast milk (HBM).

[0021] In another aspect, there is provided the use of phospholipids, their metabolic precursors and / or their metabolites for manufacturing a nutritional composition to be administered to infants and / or young children to promote and / or support an optimal myelination trajectory in the brain, which is close to the trajectory observed in infants fed only with human breast milk (HBM), and which results in promoting and / or supporting a healthy state characterized by optimal brain and cognitive function development and / or preventing neurocognitive deficits.

[0022] In another aspect, there is provided a method for promoting and / or supporting an optimal myelination trajectory in the brain, which is close to the trajectory observed in infants fed only with human breast milk (HBM), the method comprising administering to the infants and / or young children a nutritional composition comprising phospholipids, their metabolic precursors and / or their metabolites.

[0023] In another aspect, a method for promoting and / or supporting an optimal myelination trajectory in the brain is provided, the method comprising administering to an infant and / or a young child a nutritional composition comprising phospholipids, their metabolic precursors and / or their metabolites, such a trajectory being close to the trajectory observed in infants fed only with human breast milk (HBM) and leading to promoting and / or supporting a healthy state characterized by optimal brain and cognitive function development and / or preventing neurocognitive deficits.

[0024] In another aspect, the use of phospholipids, their metabolic precursors and / or their metabolites is provided for promoting and / or supporting an optimal myelination trajectory in the brain in an infant and / or a young child, such a trajectory being close to the trajectory observed in infants fed only with human breast milk (HBM).

[0025] In another aspect, the use of phospholipids, their metabolic precursors and / or their metabolites is provided for promoting and / or supporting an optimal myelination trajectory in the brain in an infant and / or a young child, such a trajectory being close to the trajectory observed in infants fed only with human breast milk (HBM) and leading to promoting and / or supporting a healthy state characterized by optimal brain and cognitive function development and / or preventing neurocognitive deficits.

[0026] The age of the infant or young child can be between 0 and 60 months, or between 0 and 24 months, or between 0 and 12 months, or between 0 and 6 months.

[0027] In another aspect of the present invention, a nutritional composition is provided, which further comprises, in addition to phospholipids (such as sphingomyelin), phospholipids, their metabolic precursors and / or their metabolites:

[0028] Fatty acid derivatives (such as DHA and / or ARA, nervonic acid and / or stearic acid), minerals (especially iron, magnesium, phosphorus, copper, calcium and / or zinc), choline, vitamin B12 and / or folic acid.

[0029] In another aspect of the present invention, a nutritional composition comprising sphingomyelin, iron, choline, DHA and folic acid is provided. Description of the Drawings

[0030] Figure 1 : Shows the myelination trajectories of infants and young children fed with breast milk compared to infants and young children fed with two commercial formula foods containing different levels of sphingomyelin. The data were obtained from the experiments described in Example 2.

[0031] Figure 2 : Shows the structure of 2-amino-4-octadecene-1,3-diol (sphingosine as defined below).

[0032] Figure 3: shows the structure of a quaternary ammonium salt containing the N,N,N-trimethylethanolammonium cation (choline as defined below).

[0033] Figure 4 : shows the effect of DHA on MBP, NF, and / or MBP / NF on day 18 and / or day 30.

[0034] Figure 5 : shows the effect of stearic acid on A2B5, MBP, MAG, NF, MBP / NF, and / or MAG / NF on day 6, day 18, and / or day 30.

[0035] Figure 6 : shows the effect of vitamin B12 on A2B5, NF, MBP / NF, and / or MAG on day 12, day 18, and / or day 30.

[0036] Figure 7 : shows the effect of folic acid on A2B5, NF, MAG, MAG / NF, and / or MBP / NF on day 12, day 18, and / or day 30.

[0037] Figure 8 : shows the effect of choline on A2B5, MAG, and / or MBP on day 12, day 18, or day 30.

[0038] Figure 9 : shows the effect of iron on A2B5, MBP, MAG, NF, and / or MAG / NF on day 12, day 18, and / or day 30.

[0039] Figure 10 : shows the effect of zinc on MBP, NF, and / or MBP / NF on day 12, day 18, and / or day 30.

[0040] Figure 11 : shows the effect of phosphorus on MAG, NF, and / or MAG / NF on day 12, day 18, and / or day 30.

[0041] Figure 12 : shows the effect of magnesium on A2B5, MBP, NF, MAG, MBP / NF, and / or MAG / NF on day 12, day 18, and / or day 30.

[0042] Figure 13 : shows the effect of copper on A2BF, MAG, and / or MAG / NF on day 12 and / or day 18.

[0043] Figure 14 : shows the effect of phosphatidylcholine on A2B5 on day 12 and on MAG on day 18.

[0044] Figure 15 : It shows the effects of phosphatidylinositol on A2B5, MBP, MAG, NF, and MAG / NF on the 12th day, 18th day, and / or 30th day.

[0045] Figure 16 : It shows the effects of phosphatidylserine on A2B5, NF, and / or MAG / NF on the 12th day and / or 18th day.

[0046] Figure 17 : It shows the effects of sphingomyelin on A2B5, MAG, and / or MBP on the 12th day, 18th day, and / or 30th day.

[0047] Figure 18 : It shows the effect of ceramide on A2B5 on the 12th day and on MAG on the 18th day.

[0048] Figure 19 : It shows the effects of galactosylceramide on A2B5, MBP, NF, and / or MBP / NF on the 12th day and / or 30th day.

[0049] Figure 20 : It shows the effect of glucosylceramide on A2B5 on the 12th day and on NF on the 12th day and 18th day.

[0050] Figure 21 : It shows the effect of D-erythro-ceramide on A2B5 on the 12th day and on MAG on the 18th day.

[0051] Figure 22 : It shows the effect of ceramide-1-phosphate on A2B5 on the 12th day and on NF and MAG on the 18th day.

[0052] Figure 23 : It shows the effects of monosialoganglioside-3 (GM3) on A2B5, MBP, MAG, and / or MBP / NF on the 12th day, 18th day, and / or 30th day.

[0053] Figure 24 : It shows the effects of disialoganglioside 3 (GD3) on A2B5, MBP, NF, and / or MAG on the 12th day, 18th day, and / or 30th day.

[0054] Figure 25 : It shows the fatty acid profiles of phosphatidylinositol (PI), phosphatidylcholine (PC), phosphatidylserine (PS), and sphingomyelin used in Example 3.

[0055] Figure 26: Shows the effects of vitamin B12 on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0056] Figure 27 : Shows the effects of ARA on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0057] Figure 28 : Shows the effects of stearic acid on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0058] Figure 29 : Shows the effects of zinc on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0059] Figure 30 : Shows the effects of phosphatidylinositol on MAG and MBP mRNA expression.

[0060] Figure 31 : Shows the effects of GD3 on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0061] Figure 32 : Shows the effects of DHA on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0062] Figure 33 : Shows the effects of nervonic acid on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0063] Figure 34 : Shows the effects of iron on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0064] Figure 35 : Shows the effects of phosphatidylcholine on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0065] Figure 36 : Shows the effects of phosphatidylserine on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0066] Figure 37 : Shows the effects of folic acid on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0067] Figure 38: Shows the effects of choline on MAG and MBP mRNA expression and on MBP and BetaIII co-expression.

[0068] Figure 39 : Shows the effects of ceramide on MAG and MBP mRNA expression and on MBP and BetaIII co-expression.

[0069] Figure 40 : Shows the effects of galactosylceramide on MAG and MBP mRNA expression and on MBP and BetaIII co-expression.

[0070] Figure 41 : Shows the effects of glucosylceramide on MAG and MBP mRNA expression and on MBP and BetaIII co-expression.

[0071] Figure 42 : Shows the effects of ceramide-1-phosphate on MAG and MBP mRNA expression and on MBP and BetaIII co-expression.

[0072] Figure 43 : Shows the effects of D-erythro-ceramide on MAG and MBP mRNA expression and on MBP and BetaIII co-expression.

[0073] Figure 44 : Shows the effects of sphingomyelin on MBP and BetaIII co-expression.

[0074] Figure 45 : Shows the effects of GM3 on MBP and BetaIII co-expression.

[0075] Figure 46 : Shows the experimental procedures of the maturation experiment.

[0076] Figure 47 : Shows the experimental procedures of the myelination experiment. Detailed implementation manners

[0077] Definition

[0078] As used herein, the following terms have the following meanings.

[0079] In the context of the present invention, the term "myelination trajectory" refers to the degree of myelination over time (related to the myelin water fraction) in infancy and early childhood.

[0080] In the context of the present invention, the term "optimal myelination trajectory" refers to the myelination trajectory as defined above that is as close as possible to the myelination trajectory achieved in infants who are exclusively breastfed during the first (3) months after birth.

[0081] If the distance between the myelination trajectory of an infant and any equivalent / same measurement point on the myelination trajectory of an infant breastfed exclusively during the first (3) months after birth is up to 50%, particularly up to 25%, more particularly up to 20%, then the myelination trajectory of the infant can be considered to be as close as possible to the myelination trajectory achieved in the infants breastfed exclusively. Non-limiting examples within the range up to 50% include 50%, 40%, 30%, 25%, 20%, 10%, 5%, 1%, 0.5% and 0.01%. In particular, the myelination trajectory will be considered to be bioequivalent.

[0082] The myelination trajectory can be measured in combination at any time point. In particular, the time point is within the first 5 years of a human life, more particularly, within the first 2 and first 3 years of a human life, even more particularly, within the first year of a human life.

[0083] The myelination trajectory can be determined by measuring the myelin-related water fraction and / or the myelin associated water pool in a subject at different time points, particularly at different time points within the first 5 years of a human subject's life, more particularly within the first 2 and first 3 years of a human life, even more particularly within the first year of a human life. The multi-component relaxation (MCR) magnetic resonance imaging (MRI) technique and particularly the mcDESPOT technique (Deoni et al 2008) can be used to measure the myelin-related water fraction and / or the myelin associated water pool in a subject. In particular, the myelination trajectory can be determined by measuring the myelin associated water pool using the mcDESPOT technique (Magn.Reson.Med. 2008 60:1372-1387, the subject matter of which is hereby incorporated herein by reference).

[0084] In the context of the present invention, the term "facilitate" means a factor or factors that cause a process to occur.

[0085] In the context of the present invention, the term "support" means a factor or factors that maintain a process once the process has started to occur.

[0086] In the context of the present invention, the expression "learning" refers to the acquisition of knowledge or skills through experience, study or by teaching.

[0087] The term "cognition" refers to the intellectual process by which an individual knows, perceives or understands ideas; thus it refers to the ability to think and understand. Cognition includes all aspects of information processing, perception, attention, thinking, reasoning, understanding and memory as well as psychomotor, language, memory, concentration, executive function and problem-solving abilities.

[0088] The term "infant" refers to a child under 12 months of age.

[0089] The expression "toddler" means a child between one and five years of age (including toddlers).

[0090] The expression "child" generally refers to humans up to 18 years of age.

[0091] A "preterm infant" or "preterm baby" is an infant or toddler born before the full term of pregnancy. Generally, it refers to an infant born before 37 weeks of gestation.

[0092] The expression "term infant" refers to an infant born after 37 weeks of gestation.

[0093] In the context of the present invention, the term "low birth weight infant" means a neonatal weight below 2500 g (5.5 pounds) due to preterm birth (i.e., before 37 weeks of gestation) and / or due to fetal growth restriction.

[0094] In the context of the present invention, the term "small for gestational age (SGA)" refers to an infant whose birth weight is below the 10th percentile of infants of the same gestational age.

[0095] The expression "postpartum period" is the period that begins immediately after birth and lasts for approximately six weeks for a child.

[0096] The expression "nutritional composition" refers to a composition that provides nutrients to a subject. Generally, a nutritional composition is ingested via the gastrointestinal tract or parenterally, or by oral or intravenous administration, and usually contains a lipid or fat source and a protein source. Preferably, the nutritional composition is for oral use.

[0097] The expression "low-allergen nutritional composition" refers to a nutritional composition that is unlikely to cause an allergic reaction.

[0098] The expression "synthetic composition" refers to a mixture obtained by chemical and / or biological methods, and the chemical properties of this mixture may be the same as those of the mixture naturally present in mammalian milk.

[0099] The expression "infant formula" refers to a food specifically for providing nutrition to infants from 4 to 6 months after birth, and which can itself meet the various nutritional needs of such infants (in accordance with Article 1.2 of Directive 91 / 321 / EEC of the European Commission of 14 May 1991 on infant formulae and follow-on formulae).

[0100] The expression "stage 1 infant formula" refers to a food specifically for providing nutrition to infants under 4 months after birth.

[0101] The term "follow-on formula" means a foodstuff which is intended for the nutrition of infants from the age of 4 months and which is the main liquid food in the increasingly diversified diet of such infants.

[0102] In the context of the present invention, the term "growing-up milk (GUM)" means a nutritional formula which can be given to children after the cessation of infant formula. "Growing-up milk" (or GUM) is used from the age of one year. It is generally a milk-based drink which is adapted to the specific nutritional needs of young children.

[0103] The term "baby food" means a foodstuff which is intended for the nutrition of infants under the age of 1 year.

[0104] The expression "fortifier" means a liquid or solid nutritional composition which is suitable for mixing with breast milk or infant formula.

[0105] The term "weaning period" means the period during which the mother's milk is gradually replaced by other foods in the infant's diet.

[0106] "Mother's milk" shall be understood to mean the mother's breast milk or colostrum (= human breast milk = HBM).

[0107] The term "fructooligosaccharide" as used herein means a fructose oligomer. It may be a long chain or a short chain, depending on the degree of polymerization of the fructooligosaccharide (the number of monomers). Preferably, the fructooligosaccharide of the present invention is a short-chain fructooligosaccharide, and most preferably it has a degree of polymerization of 2 to 10, for example a degree of polymerization of 2 to 8.

[0108] The term "sn-2 palmitate" as used herein means that palmitic acid is bonded thereto at the sn-2 position of a triglyceride.

[0109] "High sn-2 palmitic triglyceride" means a triglyceride (TG) which contains more than 30% of palmitic acid at the sn-2 position. For example, a commercially available high sn-2 palmitate component sold by Lipid Nutrition is Betapol TM B-55. It is a mixture of triglycerides derived from vegetable oils, in which at least 54% of the palmitic acid is in the sn-2 position of the glycerol molecule.

[0110] "α-Lactalbumin" means a high-quality, easily digestible whey protein which accounts for 20%-25% of the total human breast milk (HBM) protein and is the main protein present in HBM. The structure of α-lactalbumin consists of 123 amino acids and 4 disulfide bonds, and the protein has a molecular weight of 14.2K daltons. Since α-lactalbumin has a high content of essential amino acids (specifically tryptophan), it is an ideal choice for low-protein infant formulas. According to the present invention, α-lactalbumin also represents a source of sphingomyelin.

[0111] The term "prebiotic" refers to non-digestible carbohydrates that selectively stimulate the growth and / or activity of beneficial bacteria (such as Bifidobacterium in the human colon), thereby having a beneficial effect on the host (Gibson GR, Roberfroid MB. Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. J Nutr. 1995; 125: 1401-12).

[0112] The term "probiotic" refers to a preparation of microbial cells or components of microbial cells that have a beneficial effect on the health or well-being of the host. (Salminen S, Ouwehand A, Benno Y, et al. "Probiotics: how should they be defined?" Trends Food Sci. Technol. 1999; 10: 107-110). Microbial cells are generally bacteria or yeasts.

[0113] The term "sphingosine" refers to 2-amino-4-octadecene-1,3-diol having Figure 2 the structure described below. As described below, sphingosine can form the backbone of sphingolipids.

[0114] The term "sphingoid base" refers to sphingosine and a class of compounds structurally derived from it, such as dihydrosphingosine, phytosphingosine. The term "sphingoid base analogs" also includes phosphorylated forms of dihydrosphingosine, sphingosine, phytosphingosine. In one embodiment, the sphingoid base is selected from: dihydrosphingosine, sphingosine, and their phosphorylated forms.

[0115] The term "sphingolipid" refers to a class of lipids containing a sphingoid base backbone, in which the amino group (-NH2) is acylated with a fatty acid residue. "Ceramide" and "sphingomyelin" as defined below are exemplary sphingolipids.

[0116] The term "ceramide" refers to a lipid molecule in which the sphingosine backbone is acylated with a fatty acid residue. When the term ceramide is used in this specification, it can refer to a single ceramide species and a mixture of single ceramide species. In particular, ceramide is a compound of formula (IXa) or a mixture of compounds of formula (IXa)

[0117]

[0118] wherein,

[0119] R16a is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group,

[0120] and R17a is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group.

[0121] More particularly, R16a is a C13 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group which, together with the adjacent carbonyl group, corresponds to a C14 to C44 saturated or unsaturated fatty acid residue.

[0122] Non-limiting examples of C14 to C44 saturated or unsaturated fatty acids from which the fatty acid residue may be derived include: C14:0, C15:0, C16:0, C18:0, C20:0, C21:0, C22:0, C23:0, C24:1, C25:0, C28:1, C30:2, C30:1, C30:0, C32:3, C32:2, C32:1, C32:0, C33:1, C34:3, C34:2, C34:1, C34:0, C35:2, C35:0, C36:4, C36:3, C36:2, C36:1, C36:0, C37:1, C37:0, C38:4, C38:3, C38:1, C38:0, C39:1, C39:0, C40:2, C40:1, C40:0, C41:2, C41:1, C41:0, C42:47, C42:3, C42:2, C42:1, C42:0, C44:3, C44:1.

[0123] Even more particularly, R16a is a C13 to C23 branched or unbranched acyclic alkyl or acyclic alkenyl group which, together with the adjacent carbonyl group, corresponds to a C14 to C24 saturated or unsaturated fatty acid residue, wherein the fatty acid from which the fatty acid residue is derived is selected from: C14:0, C15:0, C16:0, C18:0, C20:0, C21:0, C22:0, C23:0, C24:0, C18:1n-9, C18:2n-6 and C24:1n-9, and more particularly, is selected from C16:0, C18:0, C20:0, C22:0 and C24:0.

[0124] Even more particularly, the ceramide is a mixture of compounds of formula (IXa), wherein the mixture is such that the total number of fatty acid residues (R16a taken together with the adjacent carbonyl group) contained in the mixture is predominantly saturated fatty acids and the least predominant are unsaturated fatty acids. More particularly, the mixture will be such that 80% to 96% of said fatty acid residues in the mixture are saturated fatty acids, especially C14, C15, C16, C18, C20, C22, C23, C24 saturated fatty acids, more especially C16, C18, C20, C22 and C24.

[0125] As used herein, the term "ganglioside" refers to an oligosylceramide lipid molecule containing a residue of a ceramide of formula IXa as defined herein. When the term ganglioside is used in this specification, it may refer to a single ganglioside species containing a residue of a ceramide of formula IXa as defined herein and a mixture of single ganglioside species.

[0126] The term "sphingomyelin" refers to a lipid molecule in which the sphingosine backbone is acylated at the amino group (-NH2) and in which the hydroxyl group at the 1-position of the sphingosine backbone is linked to a phospho-choline or phospho-ethanolamine group. When the term sphingomyelin is used in this specification, it may refer to a single sphingomyelin species and a mixture of single sphingomyelin species, wherein preferably the fatty acid residue is a residue of a C14 to C44 fatty acid, non-limiting examples of C14 to C44 fatty acids include C14 to C44 saturated or unsaturated fatty acids, non-limiting examples of the C14 to C44 saturated or unsaturated fatty acids from which said fatty acid residue may be derived include: C14:0, C15:0, C16:0, C18:0, C20:0, C21:0, C22:0, C23:0, C24:1, C25:0, C28:1, C30:2, C30:1, C30:0, C32:3, C32:2, C32:1, C32:0, C33:1, C34:3, C34:2, C34:1, C34:0, C35:2, C35:0, C36:4, C36:3, C36:2, C36:1, C36:0, C37:1, C37:0, C38:4, C38:3, C38:1, C38:0, C39:1, C39:0, C40:2, C40:1, C40:0, C41:2, C41:1, C41:0, C42:47, C42:3, C42:2, C42:1, C42:0, C44:3, C44:1. More preferably, the fatty acid residue is a residue of a C16, C18, C20, C22 or C24 saturated fatty acid.

[0127] The term "choline" refers to a compound containing the N,N,N-trimethylethanolammonium cation and having Figure 3Quaternary ammonium salts of the structures reported in

[0128] In the context of the present invention, the term "choline" is intended to refer to all choline present in the nutritional compositions of the present invention, whether in free form (or in the form of its salts) or in a form derived from structures containing it such as, for example: phosphatidylcholine, choline hydroxide or sphingomyelin. More preferably, the term "choline" is intended to refer to all choline present in the nutritional compositions of the present invention in free form or in the form of its salts (such as choline hydroxide).

[0129] As used herein, the term "fatty acid derivative" refers to compounds containing fatty acids other than phospholipids, and particularly to free fatty acids, and / or monoacylglycerols (hereinafter referred to as MAG) and / or diacylglycerols (hereinafter referred to as DAG) and / or triacylglycerols (hereinafter referred to as TAG) and / or cholesterol esters. More particularly, the term refers to MAG, DAG, TAG and / or cholesterol esters. Even more particularly, the term refers to TAG.

[0130] As used herein, the term "MAG" refers to a glycerol molecule in which one OH group forms an ester bond with a fatty acid. In particular, the term "MAG" as used herein refers to a compound of formula (X)

[0131]

[0132] wherein,

[0133] R 18 、R 19 or R 20 two of are H, and wherein R 18 、R 19 or R 20 one of is a C4 to C44 saturated or unsaturated acyl group.

[0134] More particularly, R 18 、R 19 or R 20 two of are H and R 18 、R 19 or R 20 one of is a C10 to C24 saturated or unsaturated acyl group, and even more particularly a C14 to C24 saturated or unsaturated acyl group.

[0135] As used herein, the term "DAG" refers to a glycerol molecule in which two of the OH groups form ester bonds with two fatty acids. In particular, the term "DAG" as used herein refers to a compound of formula (X) wherein,

[0136] R 18 、R 19 or R 20One of them is H, and wherein R 18 , R 19 or R 20 Two of them are C4 to C44 saturated or unsaturated acyl groups. More particularly C10 to C24 saturated or unsaturated acyl groups, and even more particularly C14 to C24 saturated or unsaturated acyl groups. The two C4 to C44 saturated or unsaturated acyl groups may be the same or different.

[0137] As used herein, the term "TAG" refers to a glycerol molecule in which three of the OH groups form ester bonds with three fatty acids. In particular, the term "TAG" as used herein refers to a compound of formula (X) wherein,

[0138] wherein R 18 , R 19 or R 20 are all C4 to C44 saturated or unsaturated acyl groups, more particularly C10 to C24 saturated or unsaturated acyl groups, and even more particularly C14 to C24 saturated or unsaturated acyl groups. The three C4 to C44 saturated or unsaturated acyl groups may all be the same, all different, or two may be the same and one different.

[0139] The term "cholesterol ester" as used herein refers to a compound of formula (XI)

[0140]

[0141] wherein,

[0142] R 21 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group.

[0143] More particularly, R 21 is a C9 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group, which together with the adjacent carbonyl group corresponds to a C10 to C44 saturated or unsaturated fatty acid residue, and even more particularly a C14 to C24 saturated or unsaturated fatty acid residue.

[0144] The term "fatty acid" as used herein refers to a compound of formula (XII)

[0145]

[0146] wherein

[0147] R 22 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group.

[0148] More particularly, R 22is an acyclic alkyl or alkenyl group having from C9 to C43 carbon atoms, straight-chain or branched-chain, and even more particularly an acyclic alkyl or alkenyl group having from C13 to C23 carbon atoms, straight-chain or branched-chain.

[0149] In the context of the present invention, the term "DHA" refers to docosahexaenoic acid. In the context of the present invention, the term "DHA" is intended to refer to all DHA present in the nutritional composition of the present invention, where DHA is in free form (as a fatty acid or a physiologically acceptable salt thereof) or in a form incorporated into the structure of a fatty acid derivative.

[0150] In the context of the present invention, the term "ARA" or "AA" refers to arachidonic acid. In the context of the present invention, the term "ARA" is intended to refer to all ARA present in the nutritional composition of the present invention, where ARA is in free form (as a fatty acid or a physiologically acceptable salt thereof) or in a form incorporated into the structure of a fatty acid derivative.

[0151] As used herein, the term "vitamin" refers to any vitamin. Non-limiting examples of vitamins include: vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin K, vitamin C, vitamin D, niacin, biotin, pantothenic acid, folic acid, vitamin B12, and combinations thereof.

[0152] In the context of the present invention, the term "folic acid" is intended to refer to all folic acid present in the nutritional composition of the present invention, where folic acid is in the form of itself or a physiologically acceptable salt (folate) thereof and mixtures thereof.

[0153] In the context of the present invention, the term "mineral" as used herein refers to any mineral. Non-limiting examples of minerals include: iron, zinc, calcium, phosphorus, copper, magnesium iodide, manganese, chloride, potassium, sodium, selenium, chromium, and combinations thereof. Minerals are generally added in the form of salts.

[0154] In the context of the present invention, the term "iron" is intended to refer to all iron present in the nutritional composition of the present invention, where iron is in free form, or in the form of a physiologically acceptable salt (such as, for example, ferric citrate, ferric phosphate, ferric pyrophosphate, ferrous ascorbate, ferrous carbonate, ferrous citrate, ferrous fumarate, ferrous gluconate, ferrous lactate, ferrous sulfate, or mixtures thereof), or in the form of a physiologically acceptable iron complex (such as, for example, sodium iron EDTA) and mixtures thereof.

[0155] In the context of the present invention, the term "phospholipid" as used herein refers to any phospholipid, and particularly to a compound of formula (I)

[0156]

[0157] wherein,

[0158] R 1 is O;

[0159] X is NH or O;

[0160] R 2 is a C2-C44 saturated or unsaturated, straight-chain or branched acyl group;

[0161] R 3 is a substituent of formula (II) or formula (III):

[0162] R 5 -O-CH2

[0163] (II)

[0164]

[0165] wherein R 5 is a C2-C44 saturated or unsaturated, straight-chain or branched acyl group, and

[0166] R 6 is a C2-C44 saturated alkyl or alkenyl group; and

[0167] R 4 is selected from: a C5 or C6 substituted or unsubstituted cycloalkyl or cycloalkenyl group, or

[0168] —(CH2)n—R 7 where n is an integer in the range of 1 to 4, particularly 1 to 2, and R 7 is —N(CH3)3+, NH3+ or a substituent of formula (IV), and

[0169]

[0170] In particular, R 4 is a C6 cycloalkyl or alkyl or alkenyl group substituted with one or more hydroxyl groups, more particularly R 4 is derived from inositol (C6H12O6), and even more particularly myo-inositol, i.e., R 4 is:

[0171]

[0172] As used herein, the term "acyclic" means a group that is not cyclic, i.e., does not contain a closed chain of atoms.

[0173] In the context of the present invention, the term "phosphatidylinositol" refers to a compound of formula (V)

[0174]

[0175] wherein R8 is an acyclic alkyl or alkenyl group having from C2 to C43 branches or no branches, and

[0176] R 9 is an acyclic alkyl or alkenyl group having from C2 to C43 branches or no branches.

[0177] More particularly, R 8 and R 9 are, independently of each other, acyclic alkyl or alkenyl groups having from C13 to C43 branches or no branches, which together with their adjacent carbonyl groups correspond to C14 to C44 saturated or unsaturated fatty acid residues. Even more particularly, R 8 and R 9 are, independently of each other, acyclic alkyl or alkenyl groups having from C13 to C23 branches or no branches, which together with their adjacent carbonyl groups correspond to C14 to C24 saturated or unsaturated fatty acid residues.

[0178] More particularly, R 8 and R 9 are acyclic alkyl or alkenyl groups having from C13 to C23 branches or no branches, which together with their adjacent carbonyl groups are C14 to C24 saturated or unsaturated fatty acid residues, wherein the fatty acids from which the fatty acid residues are derived are selected from: C14:0, C15:0, C16:0, C18:0, C20:0, C20:3, C20:4, C21:0, C22:0, C23:0, C24:0, C18:1n-9, C18:2n-6 and C24:1n-9. Even more particularly, selected from C18:0, C18:1n-9, C18:2, C20:3 and C20:4.

[0179] In the context of the present invention, the term "phosphatidylserine" refers to a compound of formula (VI)

[0180]

[0181] wherein R 10 is an acyclic alkyl or alkenyl group having from C2 to C43 branches or no branches, and

[0182] R 11 is an acyclic alkyl or alkenyl group having from C2 to C43 branches or no branches.

[0183] More particularly, R 10 and R 11 are, independently of each other, acyclic alkyl or alkenyl groups having from C13 to C43 branches or no branches, which together with their adjacent carbonyl groups correspond to C14 to C44 saturated or unsaturated fatty acid residues. Even more particularly, R 10 and R11 Each independently is a C13 to C23 branched or unbranched acyclic alkyl or acyclic alkenyl group, which together with their adjacent carbonyl groups corresponds to a C14 to C24 saturated or unsaturated fatty acid residue.

[0184] More particularly, R 10 and R 11 are C13 to C23 branched or unbranched acyclic alkyl or acyclic alkenyl groups, which together with their adjacent carbonyl groups are C14 to C24 saturated or unsaturated fatty acid residues, wherein the fatty acids from which the fatty acid residues are derived are selected from: C14:0, C15:0, C16:0, C18:0, C20:0, C20:3, C20:4, C21:0, C22:0, C23:0, C24:0, C18:1n-9, C18:2n-6 and C24:1n-9. Even more particularly, selected from C18:0, C18:1n-9, C20:4 and C22:6.

[0185] In the context of the present invention, the term "phosphatidylethanolamine" refers to a compound of formula (VII)

[0186]

[0187] wherein R 12 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group, and

[0188] R 13 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group.

[0189] More particularly, R 12 and R 13 each independently are C13 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl groups, which together with their adjacent carbonyl groups correspond to C14 to C44 saturated or unsaturated fatty acid residues, even more particularly, R 12 and R 13 each independently are C13 to C23 branched or unbranched acyclic alkyl or acyclic alkenyl groups, which together with their adjacent carbonyl groups correspond to C14 to C24 saturated or unsaturated fatty acid residues.

[0190] In the context of the present invention, the term "phosphatidylcholine" refers to a compound of formula (IX)

[0191]

[0192] wherein R 16 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group, and

[0193] R 17 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group.

[0194] More particularly, R 16 and R 17 are, independently of each other, C13 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl groups which, together with their adjacent carbonyl groups, correspond to C14 to C44 saturated or unsaturated fatty acid residues. Even more particularly, R 16 and R 17 are, independently of each other, C13 to C23 branched or unbranched acyclic alkyl or acyclic alkenyl groups which, together with their adjacent carbonyl groups, correspond to C14 to C24 saturated or unsaturated fatty acid residues.

[0195] More particularly, R 16 and R 17 are C13 to C23 branched or unbranched acyclic alkyl or acyclic alkenyl groups which, together with their adjacent carbonyl groups, are C14 to C24 saturated or unsaturated fatty acid residues, wherein the fatty acids from which the fatty acid residues are derived are selected from: C14:0, C15:0, C16:0, C16:1, C18:0, C20:0, C20:1, C20:3, C20:4, C21:0, C22:0, C22:6, C23:0, C24:0, C18:1n-9, C18:2n-6 and C24:1n-9. Even more particularly, selected from C14:0, C16:0, C18:0, C18:1n-9, C18:2n-6, C20:1, C20:3, C20:4 and C22:6.

[0196] The term "cfu" should be understood as colony forming units.

[0197] Breastfed infants or toddlers / Infants or toddlers fed only on breast milk: have their usual meaning and refer to infants whose major part of nutrients and / or energy is derived from human breast milk ("major part" can be at least 90% or at least 95%, or at least 99%).

[0198] Infants / toddlers fed mainly on infant formula: have their usual meaning and refer to infants whose nutritional source of nutrients and / or energy is mainly derived from synthetic infant formula, follow-on milk or growing-up milk. "Mainly" means at least 50% of those nutrients and / or energy, or at least 75%.

[0199] Unless otherwise specified, all percentages are by weight.

[0200] The present invention will now be described in more detail. It should be noted that the various aspects, features, embodiments and implementations described in this application can be compatible and / or combined together.

[0201] In addition, in the context of the present invention, the terms "comprising" or "including" do not exclude other possible elements. The compositions referred to in the present invention (including the various embodiments described herein) may comprise the following elements, consist of, or consist essentially of the following elements: the essential elements and limitations described herein, as well as any other or optional (or as required) ingredients, components, or limitations described herein.

[0202] The terms "in particular" or "more particularly" as used herein should not be considered restrictive, but should be construed as synonymous with "for example" or "especially". Detailed Description

[0204] For the sake of clarity, all embodiments and aspects reported below will apply, with the necessary modifications, to the different embodiments and aspects described for the present invention.

[0205] Unless otherwise indicated, all amounts indicated for nutrients are expressed as amounts per dry weight of the nutritional composition.

[0206] When the nutrient may be included in the composition in different forms (either in itself or in the form of salts, complexes, or more complex structures containing the nutrient), the amounts reported below are intended to refer to the amount of the nutrient itself.

[0207] The nutritional composition according to the present invention comprises phospholipids, their metabolic precursors, and / or their metabolites.

[0208] It may be particularly advantageous if the composition of the present invention further comprises one or more of the following components: vitamins and / or minerals and / or fatty acid derivatives and / or choline.

[0209] Phospholipids, their metabolic precursors, and / or their metabolites may be included in the composition in an amount up to 99.999% of the composition.

[0210] In particular, sphingomyelin, its metabolic precursors, and / or its metabolites may be included in the composition in an amount up to 99.999% of the composition.

[0211] More particularly, the composition will comprise an amount of sphingomyelin higher than 200 mg / kg dry weight of the composition, more particularly in the range of 200 mg to 2.5 g / kg dry weight of the composition.

[0212] In one embodiment, the composition comprises an amount of sphingomyelin selected from: an amount greater than 200 mg / kg, greater than 300 mg / kg, in the range of 200 mg / kg to 2.5 g / kg, in the range of 200 mg / kg to 2 g / kg, in the range of 300 mg / kg to 1.5 g / kg, or 400 mg / Kg to 1 g / Kg. All weights are based on the dry weight of the composition.

[0213] In one embodiment, phosphatidylcholine, its metabolic precursors, and / or its metabolites may be included in the composition in an amount up to 99.999% of the composition.

[0214] More particularly, the composition will comprise an amount of phosphatidylcholine greater than 200 mg / kg dry weight of the composition, more particularly in the range of 200 mg / kg to 2.5 g / kg dry weight of the composition.

[0215] In one embodiment, the composition comprises an amount of phosphatidylcholine selected from: an amount greater than 200 mg / kg, greater than 300 mg / kg, in the range of 200 mg / kg to 2.5 g / kg, in the range of 200 mg / kg to 2 g / kg, in the range of 300 mg / kg to 1.5 g / kg, or 400 mg / Kg to 1 g / Kg. All weights are based on the dry weight of the composition.

[0216] In particular, phosphatidylinositol, its metabolic precursors, and / or its metabolites may be included in the composition in an amount up to 99.999% of the composition.

[0217] More particularly, the composition will comprise an amount of phosphatidylinositol greater than 200 mg / kg dry weight of the composition, more particularly in the range of 200 mg / kg to 1.5 g / kg dry weight of the composition.

[0218] In one embodiment, the composition comprises an amount of phosphatidylinositol selected from: an amount greater than 200 mg / kg, greater than 300 mg / kg, in the range of 200 mg / kg to 2.5 g / kg, in the range of 200 mg / kg to 2 g / kg, in the range of 250 mg / kg to 800 mg / kg, or 400 mg / Kg to 1.5 g / Kg. All weights are based on the dry weight of the composition.

[0219] In particular, phosphatidylserine, its metabolic precursors, and / or its metabolites may be included in the composition in an amount up to 99.999% of the composition.

[0220] More particularly, the composition will comprise an amount of phosphatidylserine greater than 150 mg / kg dry weight of the composition, more particularly in the range of 200 mg / kg to 1.5 g / kg dry weight of the composition.

[0221] In one embodiment, the composition comprises phosphatidylserine in an amount selected from: an amount greater than 150 mg / kg, greater than 200 mg / kg, greater than 300 mg / kg, in the range of 200 mg / kg to 2.5 g / kg, in the range of 200 mg / kg to 2 g / kg, in the range of 250 mg / kg to 1000 mg / kg or 400 mg / Kg to 1 g / kg. All weights are based on the dry weight of the composition.

[0222] In particular, phosphatidylethanolamine, its metabolic precursors and / or its metabolites can be included in the composition in an amount up to 99.999% of the composition.

[0223] More particularly, the composition will comprise phosphatidylethanolamine in an amount greater than 150 mg / kg dry weight of the composition, more particularly in the range of 150 mg / kg to 1.5 g / kg dry weight of the composition.

[0224] In one embodiment, the composition comprises phosphatidylethanolamine in an amount selected from: an amount greater than 170 mg / kg, greater than 180 mg / kg, greater than 200 mg / kg, in the range of 200 mg / kg to 2.5 g / kg, in the range of 200 mg / kg to 2 g / kg, in the range of 250 mg / kg to 800 mg / kg or 200 mg / Kg to 1 g / kg. All weights are based on the dry weight of the composition.

[0225] In one embodiment, the composition of the present invention comprises phospholipids (the phospholipids include phosphatidylinositol, phosphatidylserine, phosphatidylethanolamine, sphingomyelin and phosphatidylcholine) such that the total concentration does not exceed 15.4 g / kg.

[0226] If one or more metabolic precursors and / or metabolites of phospholipids are used in the composition in place of or in combination with phospholipids, the compounds can be used in an amount such that the level of phospholipids physiologically delivered by the composition is consistent with the levels listed above. Determining the appropriate amount is within the ability of the skilled person.

[0227] The term "one or more metabolic precursors and / or metabolites of phospholipids" as used herein does not include choline.

[0228] Non-limiting examples of metabolic precursors and / or metabolites of phospholipids, particularly sphingomyelin, phosphatidylcholine, phosphatidylinositol, phosphatidylserine and / or phosphatidylethanolamine are: galactosylceramide, glucosylceramide, sphingosine, sphingosine-1-phosphate, ceramide, D-erythro-dihydroceramide and ceramide-1-phosphate and gangliosides.

[0229] Particularly effective phospholipids can be phosphatidylcholine, phosphatidylserine, phosphatidylinositol and / or sphingomyelin, particularly sphingomyelin.

[0230] In one embodiment of the present invention, the phospholipid is phosphatidylcholine, phosphatidylserine, phosphatidylinositol, sphingomyelin, and / or a metabolic precursor and / or metabolite of any of the foregoing and / or a combination of any of the foregoing. In particular, the phospholipid is sphingomyelin, its metabolic precursor, and / or its metabolite.

[0231] Particularly effective metabolic precursors and / or metabolites of phospholipids, particularly sphingomyelin, include ceramides and gangliosides, as well as ceramide-1-phosphate and d-erythro-dihydroceramide.

[0232] Particularly effective gangliosides can be monosialotetrahexosylganglioside-3 (GM3) ganglioside and / or disialotetrahexosylganglioside 3 (GD3) ganglioside.

[0233] Ceramide-1-phosphate and d-erythro-dihydroceramide will contain residues of ceramide of formula IXa as defined herein.

[0234] Gangliosides and / or ceramides and / or ceramide-1-phosphate and / or d-erythro-dihydroceramide can be included in the composition in any amount.

[0235] Concentrations in the range of 2 mg - 11.5 mg / 100 g of GD3 and / or GM3 may be particularly effective.

[0236] Sphingomyelin can be synthesized from ceramide and / or one or more gangliosides and phosphatidylcholine, and thus it may be particularly advantageous if ceramide and / or one or more gangliosides are used in combination with phosphatidylcholine, its metabolic precursor, or its metabolite.

[0237] The phospholipids, their metabolic precursors, and / or their metabolites included in the compositions of the present invention can be natural, synthetic, or mixtures thereof. The metabolic precursors and / or metabolites can be used in the compositions of the present invention in their pure or substantially pure form. As an alternative, they can be added in the form of the source containing them.

[0238] Any source of phospholipids, their metabolic precursors, and / or their metabolites suitable for ingestion by a subject intended to consume the composition can be used in the present invention.

[0239] In particular, the phospholipids, their metabolic precursors or their metabolites will be from natural sources, non-limiting examples of which include eggs, soybeans, bovine brains and / or mammalian milk or extracts thereof. Non-limiting examples of soybean sources include soy lecithin - a food additive, and non-limiting examples of mammalian milk include cow's milk, camel's milk, sheep's milk, goat's milk, including skim milk. Non-limiting extracts of milk include protein extracts (such as whey protein and casein), milk fat globule membrane (MFGM) and extracts containing them.

[0240] A particularly useful source of phospholipids, their metabolic precursors or their metabolites (especially sphingomyelin) that can be used in the present invention can be milk whey protein concentrate rich in α-lactalbumin, and / or non-pure α-lactalbumin extracted from whey protein (especially milk whey protein).

[0241] α-Lactalbumin is a high-quality, easily digestible whey protein and is the main protein present in human milk. Since α-lactalbumin and / or α-lactalbumin-rich milk fractions have a high content of essential amino acids (especially tryptophan), they are ideal for use in low-protein infant formulas. Although α-lactalbumin itself is a protein, non-pure sources may contain sphingomyelin.

[0242] In one embodiment, the phospholipids, their metabolic precursors or their metabolites (especially sphingomyelin) are used in the form of milk whey protein concentrate rich in α-lactalbumin or in the form of α-lactalbumin.

[0243] In a more specific embodiment, a milk whey protein concentrate rich in α-lactalbumin or α-lactalbumin with a phospholipid content, especially a sphingomyelin content, higher than 500 mg / 100 g of the dry weight of the composition is used.

[0244] Another particularly useful source of phospholipids, their metabolic precursors or their metabolites can be MFGM or extracts containing them, especially MFGM from milk or extracts containing them. It may be particularly advantageous if the MFGM or extracts containing them contain at least 1%, 2%, 5%, 10%, 20%, 30%, 40% of phospholipids and / or at least 0.1%, 0.2%, 0.5% to 5%, 0.8% to 3%, 1% to 2%, 1.6%, 1.9%, 1.8% of phosphatidylcholine, phosphatidylinositol, phosphatidylserine, phosphatidylethanolamine and / or sphingomyelin. MFGM may further contain magnesium, phosphorus and / or calcium, especially in a concentration range of 0.05% to 2%, 0.1% to 0.4%.

[0245] Those skilled in the art can determine the appropriate amount of the above nutrients, their metabolic precursors or their metabolites based on the nature of the composition, the purpose, the target subject and the dose, such as how many times a day the subject is to ingest the composition. Typically, the effective dose will depend on the age, size and health status of the subject, the lifestyle of the subject, the amount of nutrients in the composition and possibly on the gender of the subject.

[0246] In one embodiment, the nutritional composition according to the invention comprises sphingomyelin. In one embodiment, the nutritional composition according to the invention comprises sphingomyelin in an amount higher than 200 mg / kg.

[0247] In another embodiment, the nutritional composition of the invention comprises sphingomyelin in an amount higher than 300 mg / kg.

[0248] In yet another embodiment, the nutritional composition according to the invention comprises sphingomyelin in an amount ranging from 200 mg / kg to 2.5 g / kg.

[0249] In another embodiment, the nutritional composition according to the invention comprises sphingomyelin in an amount ranging from 200 mg / kg to 2 g / kg, such as from 300 mg / kg to 1.5 g / kg or from 400 mg / Kg to 1 g / Kg.

[0250] In the context of the present invention, sphingomyelin can be administered as such or in the form of its metabolic precursors and / or its metabolites. Without wishing to be bound by theory, it is believed that such metabolic precursors and / or metabolites have the same beneficial effects in promoting and / or supporting optimal myelination trajectories in the brain or preventing suboptimal myelination trajectories in the brain as compared to directly administering a dose of sphingomyelin. Non-limiting exemplary substances in this regard are: galactosylceramide, glucosylceramide, sphingosine, sphingosine-1-phosphate, ceramide, D-erythro-dihydroceramide and ceramide-1-phosphate and gangliosides.

[0251] The nutritional composition of the invention comprises sphingomyelin and / or its metabolic precursors or its metabolites.

[0252] Sphingomyelin is present in natural sources such as: α-lactalbumin, eggs, bovine brain or milk, etc.

[0253] Sphingomyelin and / or its metabolic precursors or its metabolites can be incorporated into the nutritional composition of the invention as a single substance, as a component consisting of a mixture of different sphingomyelin substances or by adding natural or synthetic components containing one or more sphingomyelin substances.

[0254] In one embodiment, sphingomyelin can be incorporated into the nutritional composition of the present invention in the form of being included in skim milk powder, α-lactalbumin, whey protein concentrate, and / or whey protein concentrate rich in α-lactalbumin. In one embodiment, the whey protein concentrate rich in α-lactalbumin can have a sphingomyelin content higher than 500 mg / 100 g.

[0255] In one embodiment, sphingomyelin can be incorporated into the nutritional composition of the present invention as a single component.

[0256] In one embodiment, the nutritional composition according to the present invention contains a certain amount of metabolic precursors and / or metabolites of sphingomyelin, and this amount enables them to physiologically deliver the same sphingomyelin level as that delivered by a nutritional composition containing sphingomyelin in an amount ranging from 200 mg / kg to 2.5 g / kg.

[0257] In another embodiment, the nutritional composition according to the present invention contains a certain amount of metabolic precursors and / or metabolites of sphingomyelin, and this amount enables them to physiologically deliver the same sphingomyelin level as that delivered by a nutritional composition containing sphingomyelin in an amount ranging from 400 mg / Kg to 1.5 g / Kg.

[0258] A person skilled in the art will arrive at such an amount based on their knowledge in this field.

[0259] In addition to phospholipids, their metabolic precursors and / or their metabolites, such as sphingomyelin, phosphatidylcholine, and / or phosphatidylinositol, the nutritional composition of the present invention may further contain vitamins or a mixture thereof.

[0260] Particularly effective vitamins can be folic acid, vitamin B12, and vitamin B6, especially folic acid and vitamin B12, especially folic acid.

[0261] The vitamins can be included in the composition of the present invention in an amount of 0.001% to up to 99.999% of the composition.

[0262] In one embodiment, the composition of the present invention contains vitamin B12 and / or folic acid.

[0263] Folic acid can be included in the composition of the present invention in an amount of 0.001% to up to 99.999% of the composition.

[0264] In particular, an amount of folic acid higher than 50 mcg / 100 g dry composition, more particularly 50 mcg / 100 g to 500 mcg / 100 g dry composition can be included.

[0265] In addition to phospholipids, their metabolic precursors and / or their metabolites, such as sphingomyelin, phosphatidylcholine and / or phosphatidylinositol, the nutritional composition according to the invention further comprises folic acid.

[0266] Folic acid can be included in the composition according to the invention in an amount of from 0.001% to at most 99.999% of the composition.

[0267] In one embodiment, the nutritional composition according to the invention comprises an amount of folic acid higher than 50 mcg / 100 g.

[0268] In one embodiment, the nutritional composition according to the invention comprises an amount of folic acid higher than 110 mcg / 100 g.

[0269] In one embodiment, the nutritional composition according to the invention comprises an amount of folic acid in the range of 50 mcg / 100 g to 500 mcg / 100 g or 50 mcg / 100 g to 400 mcg / 100 g.

[0270] In another embodiment, the nutritional composition according to the invention comprises an amount of folic acid in the range of 110 mcg / 100 g to 500 mcg / 100 g or 110 mcg / 100 g to 400 mcg / 100 g.

[0271] In yet another embodiment, the nutritional composition according to the invention comprises an amount of folic acid in the range of 110 mcg / 100 g to 400 mcg / 100 g or in the range of 110 mcg / 100 g to 350 mcg / 100 g.

[0272] In one embodiment, the nutritional composition according to the invention contains a level of folic acid such that the total daily intake derived from the nutritional composition according to the invention will not exceed 400 mcg.

[0273] Folic acid can be incorporated into the nutritional composition according to the invention as such or in the form of one of its physiologically acceptable salts (folates) or a mixture thereof.

[0274] In another embodiment, in addition to phospholipids, their metabolic precursors and / or their metabolites or a mixture thereof, the nutritional composition according to the invention further comprises vitamin B12.

[0275] Vitamin B12 can be included in the composition according to the invention in an amount of from 0.001% to at most 99.999% of the composition.

[0276] In particular, vitamin B12 can be included in the composition in an amount selected from: higher than 0.01 mcg, in particular higher than 0.04 mcg, in particular higher than 0.05 mcg, all weights being per 100 g of dry composition.

[0277] In one embodiment, the composition of the present invention comprises an amount of vitamin B12 selected from higher than 0.5 mcg, in the range of 0.1 mcg to 10 mcg, 0.4 mcg to 5 mcg, 0.5 mcg to 2 mcg, 1 mcg to 1.5 mcg, 4 mcg to 8.5 mcg, all weights being per 100 g of the dry composition.

[0278] In one embodiment, the composition according to the present invention comprises an amount of vitamin B12 such that the total daily intake from the nutritional composition of the present invention will not exceed 7.6 mcg / 100 g of the dry composition (77.6 mcg / Kg of the dry composition).

[0279] Vitamin B12 can be incorporated into the nutritional composition of the present invention as such or in the form of its physiologically acceptable salts or mixtures thereof or via any source containing vitamin B12. In particular, vitamin B12 can be incorporated into the composition in its pure form, as cyanocobalamin, hydroxocobalamin, and any combination thereof.

[0280] In one embodiment, in addition to phospholipids, their metabolic precursors and / or their metabolites or mixtures thereof, for example in addition to sphingomyelin, the nutritional composition according to the present invention further comprises one or more of the following components: vitamins and / or minerals and / or fatty acids and / or choline.

[0281] Particularly effective minerals can be iron, zinc, calcium, phosphorus, copper, and magnesium, especially iron.

[0282] In one embodiment, the composition of the present invention comprises iron and / or zinc and / or calcium and / or phosphorus and / or copper and / or magnesium, especially iron and zinc, more especially iron.

[0283] In another embodiment, in addition to phospholipids, their metabolic precursors and / or their metabolites, such as sphingomyelin, phosphatidylcholine, and / or phosphatidylinositol, the nutritional composition according to the present invention further comprises iron.

[0284] In one embodiment, the nutritional composition according to the present invention comprises an amount of iron higher than 5 mg / 100 g.

[0285] In one embodiment, the nutritional composition according to the present invention comprises an amount of iron higher than 9 mg / 100 g.

[0286] In one embodiment, the nutritional composition according to the present invention comprises an amount of iron in the range of 5 mg / 100 g to 40 mg / 100 g or 9 mg / 100 g to 40 mg / 100 g.

[0287] In another embodiment, the nutritional composition according to the invention comprises iron in an amount ranging from 5 mg / 100 g and 20 mg / 100 g or from 9 mg / 100 g to 20 mg / 100 g.

[0288] In another embodiment, the nutritional composition according to the invention comprises iron in an amount ranging from 5 mg / 100 g and 15 mg / 100 g or from 9 mg / 100 g to 15 mg / 100 g.

[0289] Iron can be incorporated into the nutritional composition of the invention in the form of a physiologically acceptable salt such as, for example: iron citrate, iron phosphate, iron pyrophosphate, ferrous ascorbate, ferrous carbonate, ferrous citrate, ferrous fumarate, ferrous gluconate, ferrous lactate, ferrous sulfate or mixtures thereof.

[0290] Iron can be incorporated into the nutritional composition of the invention in the form of a physiologically acceptable iron complex (such as, for example, sodium iron EDTA) or mixtures thereof.

[0291] In one embodiment, the nutritional composition according to the invention comprises a level of iron such that the total daily intake from the nutritional composition of the invention will not exceed 40 mg.

[0292] In another embodiment, in addition to phospholipids, their metabolic precursors and / or their metabolites, such as sphingomyelin, the nutritional composition according to the invention further comprises zinc.

[0293] Zinc can be included in the composition of the invention in an amount from 0.001% to at most 99.999% of the composition.

[0294] In particular, zinc can be included in the composition in an amount higher than 0.08 mg, higher than 0.3 mg, higher than 0.5 mg, all weights being per 100 g of dry composition.

[0295] In one embodiment, the composition according to the invention comprises zinc in an amount selected from the range of 0.5 mg to 8 mg, 2 mg to 5.5 mg, 2.5 mg to 4.5 mg, 3 mg to 4 mg, 4 mg to 7.5 mg, all weights being per 100 g of dry composition.

[0296] In one embodiment, the composition according to the invention comprises a level of zinc such that the total daily intake from the nutritional composition of the invention will not exceed 302.4 mg / day, or will not exceed 245 mg / day, or will not exceed 166 mg / day, or will not exceed 98.9 mg / day, or will not exceed 95.6 mg / day.

[0297] Zinc can be incorporated into the compositions of the present invention in the form of physiologically acceptable salts such as, for example: zinc nitrate, zinc sulfate, zinc gluconate, zinc acetate or mixtures thereof, or in the form of physiologically acceptable zinc complexes (such as, for example, zinc picolinate) or mixtures thereof.

[0298] In another embodiment, in addition to phospholipids, their metabolic precursors and / or their metabolites, such as sphingomyelin, the nutritional compositions according to the invention further comprise calcium.

[0299] Calcium can be comprised in the compositions of the present invention in an amount of from 0.001% to at most 99.999% of the composition.

[0300] In particular, calcium can be comprised in the composition in an amount higher than 0.84 mg, higher than 2.52 mg, higher than 4.62 mg, all weights being calculated per 100 g of dry composition of the composition.

[0301] In one embodiment, the compositions according to the invention comprise an amount of calcium in the range of from 84 mg to 760 mg, in the range of from 200 mg to 550 mg, in the range of from 250 mg to 450 mg, in the range of from 280 mg to 520 mg, from 350 mg to 650 mg, all weights being calculated per 100 g of dry composition.

[0302] In one embodiment, the compositions according to the invention comprise a level of calcium such that the total daily intake from the nutritional compositions of the present invention will not exceed 482 mg / day, or will not exceed 477 mg / day.

[0303] Calcium can be incorporated into the compositions of the present invention as such or in the form of its physiologically acceptable salts and / or via any source containing calcium. Such as calcium carbonate, calcium chloride, calcium salts of citric acid, calcium gluconate, calcium glycerophosphate, calcium lactate, calcium hydroxide, calcium salts of orthophosphoric acid.

[0304] In another embodiment, in addition to phospholipids, their metabolic precursors and / or their metabolites, such as sphingomyelin, the nutritional compositions according to the invention further comprise magnesium.

[0305] Magnesium can be comprised in the compositions of the present invention in an amount of from 0.001% to at most 99.999% of the composition.

[0306] In particular, magnesium can be comprised in the composition in an amount higher than 0.2 mg, higher than 0.35 mg, higher than 0.5 mg, all weights being calculated per 100 g of dry composition of the composition.

[0307] In one embodiment, the composition according to the invention comprises an amount of magnesium selected from the following: in the range of 0.35 mg to 90 mg, in the range of 25 mg to 70 mg, 30 mg to 65 mg, 35 mg to 60 mg, 40 mg to 50 mg, 35 mg to 55 mg, all weights being per 100 g of dry composition.

[0308] In one embodiment, the composition according to the invention comprises a level of magnesium such that the total daily intake from the nutritional composition according to the invention will not exceed 110 mg / day, or will not exceed 65 mg / day.

[0309] Magnesium can be incorporated into the composition of the invention as such or in the form of its physiologically acceptable salts and / or via any source containing magnesium, e.g., magnesium carbonate, magnesium chloride, magnesium oxide, magnesium sulfate, magnesium gluconate, magnesium hydroxide, magnesium salts of citric acid, magnesium salts of orthophosphoric acid.

[0310] In another embodiment, in addition to phospholipids, their metabolic precursors and / or their metabolites, such as sphingomyelin, the nutritional composition according to the invention further comprises phosphorus.

[0311] Phosphorus can be included in the composition of the invention in an amount of 0.001% to up to 99.999% of the composition.

[0312] In particular, phosphorus can be included in the composition in an amount higher than 1.7 mg / 100 g dry weight of the composition, higher than 14.3 mg / 100 g dry weight of the composition, higher than 27.3 mg / 100 g dry weight of the composition.

[0313] In one embodiment, the composition according to the invention comprises an amount of phosphorus selected from the following: in the range of 17 mg to 516 mg, in the range of 129 mg to 400 mg, in the range of 140 mg to 390 mg, in the range of 150 mg to 370 mg, 160 mg to 365 mg, in the range of 270 mg to 350 mg, 200 mg to 360 mg, all weights being per 100 g of dry composition.

[0314] In one embodiment, the composition according to the invention comprises a level of phosphorus such that the total daily intake from the nutritional composition according to the invention will not exceed 863 mg / day, or will not exceed 787 mg / day.

[0315] Phosphorus can be incorporated into the composition of the invention as such or in the form of its physiologically acceptable salts and / or via any source containing phosphorus, e.g., calcium phosphate, calcium hydrogen phosphate.

[0316] In another embodiment, in addition to phospholipids, their metabolic precursors and / or their metabolites, such as sphingomyelin, the nutritional composition according to the invention further comprises copper.

[0317] Copper can be included in the composition according to the invention in an amount of from 0.001% to at most 99.999% of the composition.

[0318] In particular, copper can be included in the composition in an amount higher than 10 mcg, higher than 40 mcg, higher than 60 mcg, all weights being based on the dry weight of 100 g of the composition.

[0319] In one embodiment, the composition according to the invention comprises copper in an amount selected from: higher than 100 mcg, in the range of 100 mcg to 850 mcg, 180 mcg to 650 mcg, 200 mcg to 400 mcg, 210 mcg to 300 mcg, 210 mcg to 240 mcg, 450 mcg to 850 mcg, all weights being based on the dry weight of 100 g of the composition.

[0320] In one embodiment, the composition according to the invention comprises a level of copper such that the total daily intake from the nutritional composition according to the invention will not exceed 1426 mcg / day, or will not exceed 488 mcg / day.

[0321] Copper can be incorporated into the composition according to the invention as such or in the form of its physiologically acceptable salts and / or via any source containing copper. For example, copper can be incorporated into the composition in the form of: copper sulfate and / or copper gluconate and / or copper carbonate, and / or copper citrate, and / or copper-lysine complex.

[0322] In another embodiment, in addition to phospholipids, their metabolic precursors and / or their metabolites, such as sphingomyelin, phosphatidylcholine and / or phosphatidylinositol, the nutritional composition according to the invention further comprises choline.

[0323] In one embodiment, the nutritional composition according to the invention comprises choline in an amount higher than 30 mg / 100 g.

[0324] In one embodiment, the nutritional composition according to the invention comprises choline in an amount higher than 50 mg / 100 g or higher than 100 mg / 100 g.

[0325] In one embodiment, the nutritional composition according to the invention comprises choline in an amount in the range of 30 mg / 100 g and 1000 mg / 100 g or 30 mg / 100 g to 700 mg / 100 g.

[0326] In another embodiment, the nutritional composition according to the invention comprises choline in an amount ranging from 50 mg / 100 g and 1000 mg / 100 g or from 50 mg / 100 g to 700 mg / 100 g.

[0327] In another embodiment, the nutritional composition according to the invention comprises choline in an amount ranging from 50 mg / 100 g and 500 mg / 100 g or from 100 mg / 100 g to 400 mg / 100 g.

[0328] In one embodiment, the nutritional composition according to the invention comprises a level of choline such that the total daily intake from the nutritional composition of the invention will not exceed 1 g.

[0329] Choline can be incorporated into the nutritional composition of the invention as such or in the form of a physiologically acceptable salt such as, for example: choline chloride, choline citrate, choline bitartrate or mixtures thereof. In the context of the present invention, choline can be administered in combination with sphingomyelin as such or in the form of its metabolic precursors and / or its metabolites such as, for example, phosphatidylcholine.

[0330] Choline and / or its metabolic precursors or its metabolites can be incorporated into the nutritional composition of the invention as a single substance, as a component consisting of a mixture of different choline substances or by adding a natural or synthetic component comprising one or more choline substances.

[0331] In addition to phospholipids, their metabolic precursors and / or their metabolites, such as sphingomyelin, phosphatidylcholine and / or phosphatidylinositol, the nutritional composition of the invention may further comprise choline and / or its metabolic precursors or its metabolites.

[0332] In one embodiment, the nutritional composition according to the invention further comprises a fatty acid derivative or a mixture thereof.

[0333] May be included in fatty acid derivatives, i.e., may be free fatty acids or fatty acid residues that can be derived from MAG, DAG, TAG, and / or cholesterol esters. Non-limiting examples of fatty acids with 10 to 44 carbon atoms, which can be saturated or unsaturated, include: C10:0, C12:0, C14:0, C15:0, C16:0, C16:1n-7, C18:0, C18:1n-7, C18:1n-9, C18:2n-6, 18:3n-3, C20:0, C20:1n-9, C20:2n-6, C20:3n-6, C20:4n-6, 20:5n-3, C21:0, C22:0, C22:1n-9, C22:6n-3C23:0, C24:1, especially 24:1n-9, C25:0, C28:1, C30:2, C30:1, C30:0, C32:3, C32:2, C32:1, C32:0, C33:1, C34:3, C34:2, C34:1, C34:0, C35:2, C35:0, C36:4, C36:3, C36:2, C36:1, C36:0, C37:1, C37:0, C38:4, C38:3, C38:1, C38:0, C39:1, C39:0, C40:2, C40:1, C40:0, C41:2, C41:1, C41:0, C42:47, C42:3, C42:2, C42:1, C42:0, C44:3, C44:1. In particular, the fatty acids will be selected from: C10:0, C12:0, C14:0, C16:0, C16:1n-7, C18:0, C18:1n-7, C18:1n-9, C18:2n-6, 18:3n-3, C20:0, C20:1n-9, C20:2n-6, C20:3n-6, C20:4n-6, 20:5n-3, C22:0, C22:1n-9, C22:6n-3, C24:1, 24:1n-9, especially 24:1n-9.

[0334] It may be particularly advantageous if the fatty acid derivative contains a saturated or unsaturated fatty acid selected from C20:4n-6, C22:6n-3, C24:1n-9, C16:0, C18:1n-9, and C18.0. In particular, C20:4n-6 and / or C22:6n-3 and / or C18:0. More particularly 22:6n-3 and / or C18:0.

[0335] Any fatty acid derivative suitable for ingestion by a subject intended to consume the composition can be used in the present invention.

[0336] In particular, the fatty acid derivatives will be from natural sources, non-limiting examples of which include eggs, algae, fish oil, molds, yeast, seeds, plants such as soybeans, and animal sources such as bovine brain, and / or mammalian milk or extracts thereof. Non-limiting examples of soybean sources include soy lecithin - a food additive, and non-limiting examples of mammalian milk include cow milk, camel milk, sheep milk, goat milk, including skim milk. Non-limiting extracts of milk include protein extracts, milk fat globule membrane (MFGM), and extracts containing them. Fatty acid derivatives can also be from palm oil, beef tallow, lard, cottonseed oil, peanut oil.

[0337] It may be particularly advantageous if the fatty acid derivative contains a saturated or unsaturated fatty acid selected from C20:4n-6, C22:6n-3, C24:1n-9, C16:0, C18:1n-9, and C18.0. In particular C20:4n-6 and / or C22:6n-3 and / or C18:0. More particularly 22:6n-3 and / or C18:0.

[0338] C20:4n-6 is arachidonic acid (hereinafter referred to as ARA or AA). C22:6n-3 is docosahexaenoic acid (hereinafter referred to as DHA). 24:1n-9 is nervonic acid. C18.0 is stearic acid. C16:0 is palmitic acid. C18:1n-9 is oleic acid.

[0339] In one embodiment, the composition according to the invention contains a fatty acid derivative comprising DHA and / or ARA and / or nervonic acid and / or stearic acid, particularly a fatty acid derivative comprising DHA and / or ARA and / or stearic acid. Most particularly a fatty acid derivative comprising DHA and / or stearic acid.

[0340] The fatty acid derivative comprising DHA and / or ARA and / or nervonic acid and / or stearic acid can be included in the composition according to the invention in an amount of up to 99.999% of the composition.

[0341] In particular, the fatty acid derivative comprising DHA and / or ARA and / or nervonic acid and / or stearic acid can be included in the composition according to the invention in an amount of 15 mg / 100 g to 350 mg / 100 g of the dry weight of the composition, more particularly 30 mg / 100 g to 300 mg / 100 g of the dry weight of the composition.

[0342] In one embodiment, the composition according to the invention contains a fatty acid derivative comprising DHA and / or ARA and / or nervonic acid and / or stearic acid in an amount selected from: above 15 mg / 100 g dry weight of the composition, above 30 mg / 100 g dry weight of the composition, above 50 mg / 100 g dry weight of the composition, in the range of 30 mg / 100 g dry weight to 300 mg / 100 g dry weight of the composition, in the range of 30 mg / 100 g dry weight to 200 mg / 100 g dry weight of the composition or 30 mg / 100 g dry weight to 150 mg / 100 g dry weight of the composition, in the range of 50 mg / 100 g dry weight to 300 mg / 100 g dry weight of the composition, in the range of 50 mg / 100 g dry weight to 200 mg / 100 g dry weight of the composition, in the range of 50 mg / 100 g dry weight to 150 mg / 100 g dry weight of the composition.

[0343] Fatty acid derivatives containing stearic acid are present in natural sources such as palm oil, beef tallow, lard, cottonseed oil, and peanut oil.

[0344] Fatty acid derivatives containing nervonic acid are present in natural sources such as the seed oils of Cardamine gracea, Heliphila longifola, Thlaspi perfoliatum, Tropaeolum speciosum, Lunaria biennis, Lunaria annua, and Malania oleifera; the molds Neocallismastix frontalis, Erysiphe graminis, and Sphaerotheca humuli; the bacterium Pseudomonas atlantica; the yeast Saccharomyces cerevisiae; and the marine diatom Nitzschia cylindrus.

[0345] Fatty acid derivatives containing DHA and / or ARA are present in natural sources such as eggs, algae, fungi, or fish oil.

[0346] Oils containing fatty acid derivatives comprising DHA and / or ARA and usually other polyunsaturated fatty acids (PUFAs), in particular EPA (eicosapentaenoic acid), can have various sources. Preferably, the fatty acid derivative containing DHA is provided in the form of fish oil containing fatty acid derivatives comprising DHA and / or ARA. Fish oil usually contains 5% by weight or more, preferably 10% by weight or more, of fatty acid derivatives comprising DHA and / or ARA. Oils containing a large amount of fatty acid derivatives comprising DHA and / or ARA obtained from algae or microorganisms are also usually available. For example, an oil harvested from algae containing 10% by weight or more, such as 20% by weight or more, of fatty acid derivatives can be used.

[0347] If the nutritional composition according to the invention contains fatty acid derivatives comprising ARA and DHA. The ingredient can be included in the composition of the invention, for example, in an amount such that the weight ratio of DHA:ARA is in the range of 4:1 to 1:4, such as 3:1 to 1:3, such as 2:1 to 1:2, such as 1.5:1 to 1:1.5, especially 1.1:1 to 1:1.1.

[0348] It may also be advantageous if the composition of the invention contains a mixture of fatty acid derivatives, wherein the mixture is such that the weight ratio of unsaturated fatty acids to saturated fatty acids and / or fatty acid residues in the composition of the invention is in the range of 1:1 to 1:2, 1:1.2 to 1:1.9, 1:1.25 to 1:1.5, 1:3 to 1:4.

[0349] Furthermore, when a large amount of fatty acid derivatives comprising DHA and / or ARA are included in the composition of the invention, it may be particularly advantageous if the total amount of saturated long-chain fatty acids, especially fatty acid derivatives of C20 / 24, is increased. These saturated long-chain fatty acids may be important components of myelin, enabling it to wrap and encapsulate axons. The weight ratio of DHA and / or AA to these unsaturated long fatty acids in the composition of the invention can be, for example, in the range of 1:1 to 1:10, 1:2 to 1:9, 1:3 to 1:4.5, 1:3.5 to 1:4.5.

[0350] The nutritional composition according to the invention contains ARA.

[0351] In one embodiment, the nutritional composition according to the invention contains an amount of ARA higher than 30 mg / 100 g.

[0352] In one embodiment, the nutritional composition according to the invention contains an amount of ARA higher than 50 mg / 100 g.

[0353] In one embodiment, the nutritional composition according to the invention comprises ARA in an amount ranging from 30 mg / 100 g to 300 mg / 100 g or from 30 mg / 100 g to 200 mg / 100 g or from 30 mg / 100 g to 150 mg / 100 g.

[0354] In one embodiment, the nutritional composition according to the invention comprises ARA in an amount ranging from 50 mg / 100 g to 300 mg / 100 g or from 50 mg / 100 g to 200 mg / 100 g or from 50 mg / 100 g to 150 mg / 100 g.

[0355] ARA is present in natural sources such as eggs, fungi, algae or fish oil.

[0356] According to one embodiment, ARA is provided in the form of triglycerides containing ARA.

[0357] Oils containing ARA and usually other polyunsaturated fatty acids (PUFAs), in particular EPA (eicosapentaenoic acid), can have various sources. Preferably, ARA is provided in the form of fish oil containing ARA. Fish oil usually contains 5% by weight or more, preferably 10% by weight or more of ARA. Oils containing a large amount of ARA obtained from algae or microorganisms are also available for use. For example, oils harvested from fungi containing 10% by weight or more, such as 20% by weight or more of ARA, can be used.

[0358] ARA can be incorporated into the nutritional composition of the invention as a single substance (as a fatty acid, in the form of its physiologically acceptable salts or in the form of triglycerides containing it), as a component consisting of a mixture of different ARA substances, or by adding natural or synthetic components containing one or more ARA substances.

[0359] In one embodiment, in addition to ARA, the nutritional composition of the invention further comprises: phospholipids (especially sphingomyelin), minerals (especially iron, magnesium, phosphorus, calcium and / or zinc), choline, DHA, vitamin B12 and / or folic acid.

[0360] In another embodiment, in addition to sphingomyelin, the nutritional composition according to the invention further comprises DHA.

[0361] In one embodiment, the nutritional composition according to the invention comprises an amount of DHA higher than 30 mg / 100 g.

[0362] In one embodiment, the nutritional composition according to the invention comprises an amount of DHA higher than 50 mg / 100 g.

[0363] In one embodiment, the nutritional composition according to the invention comprises DHA in an amount ranging from 30 mg / 100 g and 300 mg / 100 g or 30 mg / 100 g and 200 mg / 100 g or from 30 mg / 100 g to 150 mg / 100 g.

[0364] In one embodiment, the nutritional composition according to the invention comprises DHA in an amount ranging from 50 mg / 100 g to 300 mg / 100 g or 50 mg / 100 g to 200 mg / 100 g or from 50 mg / 100 g to 150 mg / 100 g.

[0365] The nutritional composition of the invention comprises DHA.

[0366] DHA is present in natural sources such as eggs, algae or fish oil.

[0367] According to one embodiment, DHA is provided in the form of triglycerides containing DHA.

[0368] Oils containing DHA and usually other polyunsaturated fatty acids (PUFAs), especially EPA (eicosapentaenoic acid), can have various sources. Preferably, DHA is provided in the form of fish oil containing DHA. Fish oil usually contains 5% by weight or more, preferably 10% by weight or more of DHA. Oils containing a large amount of DHA obtained from algae or microorganisms can also be used. For example, oils harvested from algae containing 10% by weight or more, such as 20% by weight or more of DHA, can be used.

[0369] DHA can be incorporated into the nutritional composition of the invention as a single substance (as a fatty acid, in the form of its physiologically acceptable salts or in the form of triglycerides containing it), as a component consisting of a mixture of different DHA substances or by adding natural or synthetic components containing one or more DHA substances.

[0370] In one embodiment, the nutritional composition according to the invention comprises arachidonic acid (ARA) and DHA.

[0371] According to one embodiment, the nutritional composition comprises approximately the same amount of DHA and ARA. For example, the weight ratio of DHA:ARA ranges from 4:1 to 1:4, such as 3:1 to 1:3, such as 2:1 to 1:2, such as 1.5:1 to 1:1.5, especially in the range of 1.1:1 to 1:1.1.

[0372] In one embodiment, when the nutritional composition contains a large amount of DHA, the unsaturated long-chain fatty acids c20 / 22 / 24 increase.

[0373] In one embodiment, the nutritional composition according to the invention comprises sphingomyelin, iron and choline.

[0374] In another embodiment, the nutritional composition according to the present invention comprises sphingomyelin, iron, and DHA.

[0375] In another embodiment, the nutritional composition according to the present invention comprises sphingomyelin, iron, and folic acid.

[0376] In one embodiment, the nutritional composition according to the present invention comprises sphingomyelin, DHA, and choline.

[0377] In another embodiment, the nutritional composition according to the present invention comprises sphingomyelin, DHA, and iron.

[0378] In another embodiment, the nutritional composition according to the present invention comprises sphingomyelin, DHA, and folic acid.

[0379] In one embodiment, the nutritional composition according to the present invention comprises sphingomyelin, choline, and DHA.

[0380] In another embodiment, the nutritional composition according to the present invention comprises sphingomyelin, choline, and iron.

[0381] In another embodiment, the nutritional composition according to the present invention comprises sphingomyelin, choline, and folic acid.

[0382] In one embodiment, the nutritional composition according to the present invention comprises sphingomyelin, folic acid, and DHA.

[0383] In another embodiment, the nutritional composition according to the present invention comprises sphingomyelin, folic acid, and iron.

[0384] In another embodiment, the nutritional composition according to the present invention comprises sphingomyelin, folic acid, and choline.

[0385] In one embodiment, the nutritional composition according to the present invention comprises sphingomyelin, folic acid, iron, and DHA.

[0386] In another embodiment, the nutritional composition according to the present invention comprises sphingomyelin, folic acid, choline, and iron.

[0387] In another embodiment, the nutritional composition according to the present invention comprises sphingomyelin, folic acid, DHA, and choline.

[0388] In another embodiment, the nutritional composition according to the present invention comprises sphingomyelin, iron, DHA, and choline.

[0389] In one embodiment, the nutritional composition according to the present invention comprises sphingomyelin, folic acid, iron, choline, and DHA.

[0390] In one embodiment, the composition according to the invention comprises: fatty acid derivatives, in particular fatty acid derivatives comprising DHA and / or ARA and / or nervonic acid and / or stearic acid; and vitamins, in particular B12 and / or folic acid; and / or phospholipids, in particular phosphatidylcholine, and / or phosphatidylserine, and / or phosphatidylinositol, and / or sphingomyelin, and / or metabolic precursors or metabolites of any of the foregoing; and / or minerals, in particular iron, and / or zinc, and / or calcium, and / or phosphorus, and / or magnesium; and / or choline.

[0391] Particularly beneficial concentrations / amounts of the components in the composition may be a sphingomyelin amount of at least 420 mg / kg, a phosphatidylcholine amount of at least 1000 mg / kg, a phosphatidylserine amount of at least 900 mg / kg, a phosphatidylinositol amount of at least 700 mg / kg, a folic acid amount of at least 160 mg / kg, a vitamin B12 amount of at least 7 mcg / 100 g, an iron amount of at least 11.5 mg / 100 g, a choline amount of at least 140 mg / kg, a fatty acid derivative comprising DHA amount of at least 89 mg / 100 g, a fatty acid derivative comprising AA amount of at least 175 mg / 100 g, a zinc amount of at least 7 mg / 100 g, a calcium amount of at least 500 mg / 100 g, a phosphorus amount of at least 350 mg / 100 g, a copper amount of at least 600 mcg / 100 g, a magnesium amount of at least 50 mg / 100 g. All weights are based on the dry weight of the composition.

[0392] In one embodiment, the composition according to the invention contains phospholipids and / or their metabolic precursors or their metabolites (e.g., sphingomyelin, phosphatidylcholine and / or phosphatidylinositol), fatty acid derivatives (e.g., comprising DHA and / or ARA), vitamin B12 and / or folic acid, iron and choline.

[0393] In one embodiment, the composition according to the invention contains fatty acid derivatives comprising DHA and / or ARA, vitamin B12 and / or folic acid, sphingomyelin and iron.

[0394] In a more specific embodiment, the composition according to the invention contains fatty acid derivatives comprising DHA at a concentration of 1023 mg / kg, ARA at a concentration of 1023 mg / kg, vitamin B12 at a concentration of 54 mcg / kg, folic acid at a concentration of 1698 mcg / kg, sphingomyelin at a concentration of 814 mg / kg and iron at a concentration of 67 mg / kg.

[0395] The composition of the invention can be any type of composition suitable for direct administration to a subject.

[0396] In particular, the composition will be a synthetic nutritional composition.

[0397] Those skilled in the art will determine the appropriate amounts of the above nutrients, their metabolic precursors or their metabolites to achieve their maximum allowable levels in the nutritional composition after administration.

[0398] Preferred nutritional composition matrix :

[0399] The composition according to the invention can be a synthetic nutritional composition. It can be an infant formula, a stage 1 infant formula, a follow-on formula, a growing-up milk, a baby food, a preterm formula or a fortifier such as a human milk fortifier or a supplement. Preferably, the composition of the invention is an infant formula or a fortifier or a supplement dedicated to infants less than 4 months or less than 6 months old.

[0400] In a typical embodiment of the invention, the composition will contain a protein source, a lipid source and a carbohydrate source.

[0401] For example, such a composition may contain from about 2 g / 100 kcal to 6 g / 100 kcal of protein, from about 1.5 g / 100 kcal to 3 g / 100 kcal of lipid and / or from about 1.7 g / 100 kcal to 12 g / 100 kcal of carbohydrate.

[0402] If the composition is a liquid, its energy density can be between 60 kcal / 100 ml and 75 kcal / 100 ml.

[0403] If the composition is a solid, its energy density can be between 60 kcal / 100 g and 75 kcal / 100 g.

[0404] The type of protein is considered immaterial to the present invention. Thus, protein sources based on, for example, whey, casein and mixtures thereof can be used. In the case of whey proteins, acid whey or sweet whey or mixtures thereof can be used, as well as α-lactalbumin and β-lactoglobulin in any desired proportions. The whey protein can be modified sweet whey. Sweet whey is a by-product of the cheese-making process that is more readily available and is commonly used in the manufacture of milk-based infant formula. However, sweet whey contains a component called caseinomacropeptide (CGMP), which is undesirably rich in threonine and low in tryptophan. Removing CGMP from sweet whey can bring the threonine content of the protein closer to that in human milk. Then those amino acids that are present in low amounts (primarily histidine and tryptophan) can be supplemented in this modified sweet whey. A method for removing CGMP from sweet whey is described in EP 880902, and an infant formula based on such modified sweet whey is described in WO 01 / 11990. The protein can be whole or hydrolyzed, or a mixture of whole and hydrolyzed proteins. For example, for subjects considered to be at risk of developing a milk allergy, it may be desirable to provide a partially hydrolyzed protein (degree of hydrolysis between 2% and 20%). If a hydrolyzed protein is required, the hydrolysis process can be carried out as needed and as known in the art. For example, the whey fraction can be enzymatically hydrolyzed in two steps as described in EP 322589 to prepare a whey protein hydrolysate. For a fully hydrolyzed protein, the whey protein can be triple-hydrolyzed using first Alcalase 2.4L (EC 940459) at 55°C, then Neutrase 0.5L (available from NovoNordisk Ferment AG), and then trypsin. If the whey fraction used as a raw material is substantially lactose-free, it has been found that the protein undergoes much less lysine blockage during the hydrolysis process. This makes it possible to reduce the degree of lysine blockage from about 15% by weight of the total lysine to less than about 10% by weight of lysine; for example, about 7% by weight of lysine, which greatly improves the nutritional quality of the protein source.

[0405] The compositions of the present invention can contain a source of carbohydrates. Any source of carbohydrates can be used, such as lactose, sucrose, maltodextrin, starch, honey, and mixtures thereof.

[0406] The compositions of the present invention may contain a lipid source. The lipid source can be any lipid. Preferred fat sources include milk fat and vegetable oils (including but not limited to palm oil, high oleic sunflower oil, and high oleic safflower oil). Essential fatty acids, namely linoleic acid and α-linolenic acid, can also be added. In one embodiment, a small amount of oil rich in crude arachidonic acid (AA) and docosahexaenoic acid (DHA), such as fish oil or microbial oil, can be added. The ratio of n-6 fatty acids to n-3 fatty acids in the lipid source is preferably from about 5:1 to about 15:1; for example, from about 8:1 to about 10:1.

[0407] The compositions of the present invention may also contain all vitamins and minerals known to be essential in the daily diet and highly demanded nutritionally. The minimum requirements for certain vitamins and minerals have been determined. Examples of minerals, vitamins, and other nutrients optionally present in infant formula include vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin E, vitamin K, vitamin C, vitamin D, folic acid, inositol, niacin, biotin, pantothenic acid, choline, calcium, phosphorus, iodine, iron, magnesium, copper, zinc, manganese, chloride, potassium, sodium, selenium, chromium, molybdenum, taurine, and L-carnitine. Minerals are usually added in the form of salts. In one embodiment, the composition contains an amount of vitamin B12 and / or folic acid. The presence and amount of specific minerals and other vitamins will vary depending on many factors, such as the age, weight, and condition of the human or animal to whom the composition is to be administered.

[0408] The composition may also contain at least one probiotic strain. Probiotics are preparations of microbial cells or components of microbial cells that have a beneficial effect on the health or well-being of the host. Suitable probiotic strains include: Lactobacillus rhamnosus ATCC 53103, Lactobacillus rhamnosus CGMCC 1.3724, Lactobacillus paracasei CNCM I-2116, which can be purchased from Valio Oy, Finland, under the trademark LGG, Bifidobacterium lactis CNCM I-3446, sold especially by the Christian Hansen company under the trademark Bb12, and Bifidobacterium longum ATCC BAA-999, sold by Morigana Milk Industry Co., Ltd., Japan, under the trademark BB536. If a probiotic is present, its amount also preferably varies according to the age of the human or animal. Generally speaking, the probiotic content can increase with the age of the infant, for example, 10 3cfu / g formula food (dry weight) up to 10 12 cfu / g formula food (dry weight), more preferably between 10 4 cfu / g formula (dry weight) and 10 8 cfu / g formula food (dry weight).

[0409] The composition may further comprise at least one prebiotic in an amount of 0.3% to 10%. Prebiotics are indigestible food ingredients that beneficially affect the host by selectively stimulating the growth and / or activity of one or a limited number of bacteria in the colon, thereby improving the host's health. The fact that these ingredients are indigestible means that they cannot be broken down and absorbed in the stomach or small intestine, so they can pass through the stomach and small intestine intact and reach the colon, where they are selectively fermented by beneficial bacteria. Examples of prebiotics include certain oligosaccharides.

[0410] The composition may optionally comprise other substances that may have beneficial effects, such as nucleotides, nucleosides, etc.

[0411] The compositions used in the present invention, such as infant formula foods, can be prepared by any suitable means. For example, an infant formula food can be prepared by mixing a protein source, a carbohydrate source, and a fat source in suitable proportions. If an emulsifier is used, it can be included in the mixture. Vitamins and minerals can be added at this time, but are usually added a little later to avoid thermal degradation. Any lipophilic vitamins, emulsifiers, etc. can be dissolved in the fat source first and then mixed. Then water (preferably reverse osmosis water) can be incorporated to form a liquid mixture. Subsequently, the liquid mixture can be heat-treated to reduce the bacterial count. For example, the liquid mixture can be rapidly heated to a temperature in the range of about 80°C to about 110°C and held for about 5 seconds to about 5 minutes. This can be carried out by steam injection or through a heat exchanger (such as a plate heat exchanger). Then the liquid mixture can be cooled to about 60°C to about 85°C, for example, by rapid cooling. Then the liquid mixture can be homogenized; for example, in two stages: the first stage is carried out at about 7 MPa to about 40 MPa, and the second stage is carried out at about 2 MPa to about 14 MPa. Then the homogenized mixture can be further cooled to add any heat-sensitive components; such as vitamins and minerals. At this point, it is convenient to standardize the pH and solids content of the homogenized mixture. The homogenized mixture is transferred to a suitable drying device (such as a spray dryer or a freeze dryer) and converted into a powder. The water content of the powder should be less than about 5% by weight. If it is desired to add one or more probiotics, the probiotics can be cultured according to any suitable method and then, for example, made by freeze drying or spray drying for addition to the infant formula food. Alternatively, bacterial products that have been made into a suitable form can be purchased from professional suppliers such as Christian Hansen of Denmark and Morinaga of Japan for addition to food products (such as infant formula foods). Such bacterial products can be added to the powdered infant formula food by dry mixing.

[0412] Phospholipids, their metabolic precursors, and / or their metabolites (such as sphingomyelin) can be added at any stage in the process, but are preferably added after the heating step.

[0413] In one embodiment, the nutritional composition comprises triglycerides having a high sn-2 palmitate, preferably triglycerides having more than 33% of palmitic acid at the sn-2 position.

[0414] In some embodiments, by weight, palmitic acid accounts for about 15% to about 25% of the total fatty acid content of the formula food, such as about 15% to about 20%, and at least about 30% (e.g., about 35%) to about 43% of the total palmitic acid content is located at the sn-2 position.

[0415] A commercially available composition sold by Lipid Nutrition is Betapol TM B-55, which is a mixture of triglycerides derived from vegetable oils, in which at least 54% of the palmitic acid is in the sn-2 position of the glycerol molecule. In one embodiment, the fat content of the composition of the present invention is about 40% to 50% by weight (e.g., about 43% to about 45% by weight) of Betapol TM B-55. Those skilled in the art will appreciate that without departing from the essence and scope of the present invention, the percentage of high sn-2 fat used in the formulated food and the total amount of sn-2 palmitate can vary, and different high sn-2 palmitate oils can be used.

[0416] Health effect

[0417] The composition of the present invention has a positive effect on the myelination trajectory in the brain of infants or young children fed with this composition.

[0418] This positive effect can include promoting and / or supporting an optimal myelination trajectory in the subjects fed with the composition, which determines the proper development of cognitive skills and abilities and learning in infants or young children. The optimal myelination trajectory can also prevent the occurrence of cognitive impairment or delay it.

[0419] Health effects can be observed after using a nutritional composition containing phospholipids, their metabolites, and / or their metabolic precursors (such as sphingomyelin, phosphatidylcholine, and / or phosphatidylinositol) for several days, weeks, or months.

[0420] The effect of the present invention can be preventive (e.g., avoiding a suboptimal myelination trajectory in the brain) or curative (restoring an optimal myelination trajectory in the brain).

[0421] The health effects associated with the infant can be measured by various methods as shown in the following examples.

[0422] Target infant

[0423] In one embodiment of the present invention, the target infant or young child shows a suboptimal myelination trajectory in the brain, which can lead to cognitive defects, impaired cognitive abilities, and / or suboptimal cognitive development.

[0424] In one embodiment, such an infant can be a premature infant, a low birth weight infant, or a small for gestational age infant.

[0425] In another embodiment, the infant or young child is born at term. All infants can benefit from the present invention because all infants are prone or may be prone to developing a suboptimal myelination trajectory in the brain.

[0426] In such infants or toddlers, it is of particular interest to obtain a myelination trajectory in the brain that is close to that of breastfed infants (preferably infants breastfed only for the first few months after birth). In fact, it provides them with a state of health in terms of cognitive abilities that is consistent with that observed in breastfed infants.

[0427] In one embodiment, the infants and toddlers are 0 to 3 months old, 0 to 6 months old, or 0 to 12 months old or 0 to 36 months old or 0 to 60 months old. It is foreseeable that the compositions of the present invention may be more beneficial when administered to infants shortly after birth (0 - 4 weeks, 0 - 8, 0 - 12, 0 - 24 weeks), as the myelination process has begun and is significantly developed at this time.

[0428] The following examples are presented to illustrate certain embodiments and features of the present invention, but should not be construed as limiting the scope of the invention.

[0429] Expected feeding regimen

[0430] In one embodiment, the composition of the present invention is fed to an infant or toddler (or intended to be fed or directed to be fed) for 2 to 52 weeks. In one embodiment, it is fed to an infant or toddler for 2 to 24 weeks, or 2 to 12 weeks.

[0431] In one embodiment, the composition of the present invention is fed to an infant or toddler (or intended to be fed or directed to be fed) for 2 to 52 weeks and is started shortly after the birth of the infant or the interruption of breastfeeding. In one embodiment, the composition of the present invention is fed to an infant or toddler for 2 to 24 weeks, or 2 to 12 weeks and is started shortly after the birth of the infant or the interruption of breastfeeding.

[0432] Without wishing to be bound by theory, it is believed that starting early (at birth or shortly after birth) is preferred to induce the desired effect.

[0433] It is expected that when the composition of the present invention is used as the sole source of nutrition (in addition to supplementing breastfeeding), the health benefits are more prominent or are established more quickly. In one embodiment, the health effects are observed as long as the composition of the present invention is used to cover 50% or more, or 75% or more of the nutritional requirements (such as energy requirements) of the target infant or toddler.

[0434] Experimental section :

[0435] Methods, definitions, and materials

[0436] MRI (Magnetic Resonance Imaging) :

[0437] MRI brain scans of infants and children between 0 and 5 years of age were obtained using white matter imaging techniques. This technique provides a quantitative measure, the myelin water fraction (MWF), which is a surrogate marker for myelin content. When plotted over time throughout early childhood, a myelination trajectory can be generated.

[0438] Infant formula composition : Analyze the composition of six infant formulas fed to the infants participating in the study / the levels of myelin-related nutrients.

[0439] The test compositions were standard commercial infant formulas from different brands / suppliers and showed different levels of nutrients contained therein.

[0440] Cognitive ability : Age-standardized (T) scores for gross motor, visual reception, and language (expressive and receptive) were derived from the Mullen Scales of Early Learning.

[0441] Clinical study

[0442] Infant participants

[0443] The infants included in this study were drawn from a larger longitudinal study of normal brain and behavioral development: the Brown university Assessment of Myelination and Behavior Across Maturation (BAMBAM). To focus on typical neurodevelopment, children known to have potential risk factors for learning disabilities, neurological disorders, or mental disorders were explicitly excluded at the time of recruitment and inclusion. Thus, children with in utero alcohol or illicit substance exposure, preterm birth (<37 weeks of gestation) or multiple births, fetal ultrasound abnormalities, concurrent pregnancy (such as preeclampsia), APGAR score <8, admission to the neonatal intensive care unit, neurological disorders (such as head injury, epilepsy), or a mental disorder or developmental disorder in the infant, parent, or sibling (including maternal depression requiring medication) were excluded. Continuous screening (such as the MCAT for autism, or the CBCL for behavioral problems) was further used to exclude enrolled children with clinically relevant behavior or obvious medical conditions (such as autism spectrum disorder).

[0444] The combination of retrospective and prospective data was obtained from parents through a detailed medical history and interviews of parents regarding the type of infant formula used, the proportion of breastfed to formula-fed infants, and the duration of exclusive breastfeeding. This information was updated at each study visit, which occurred approximately every 6 months for children under 2 years of age and annually for older children. Using this information, children were classified into one of two groups: Group 1, formula-fed only; Group 2, breastfed exclusively for at least 90 days (3 months). Children who were fed a combination of breast milk and formula within 3 months were excluded from our analysis. Infants in the formula-fed only group were further sub-classified based on the primary infant formula used during the first three months as reported by the parents. The primary formula was defined as the formula given 90% or more of the time (e.g., in cases where parents used an alternative brand during a vacation).

[0445] Using these criteria, 94 formula-fed infants and toddlers were selected for Group 1. This included 13 children who received Formula #2; 28 children who received Formula #5; 8 children who received Formula #3; 39 children who received Formula #4; 5 children who received Formula #1; and 1 child who received Formula #6. A sample of 52 breastfed infants was also selected and matched to the formula-fed group with respect to mean age, gestational age, birth weight, male-to-female ratio, racial proportion, maternal education level, family size, and number of languages used in the home (other than English). Group demographics are provided in Table 1.

[0446] Table 1: Data breakdown for longitudinal and nutritional analysis

[0447]

[0448] Imaging methods and analysis

[0449] Each infant was scanned using the mcDESPOT (Multicomponent driven-equilibrium single-pulse observation of T1 and T2) white matter imaging technique of Deoni et al. (Magn. Reson. Med. 2008, 60:1372 - 1387), which provides a quantitative measure of the myelin water fraction (MWF) at each imaging point throughout the brain. All infants were scanned using a noise-attenuated mcDESPOT imaging protocol during natural (i.e., non-sedated) sleep. Total imaging time varied from 19 minutes for the youngest toddlers to 24 minutes for older 4-year-old children.

[0450] All data were acquired on a Siemens 3T Tim Trio scanner equipped with a 12-channel head RF array. To minimize movement during scanning, children were restrained with a pediatric MedVac vacuum immobilization bag (CFI Medical Solutions, USA) and foam pads. Scanner noise was reduced by decreasing the peak gradient amplitude and slew rate and using a soundproof scanner bore insert (Quiet Barrier HD Composite, UltraBarrier, USA). MiniMuff pediatric ear muffs and electro-dynamic headphones (MR Confon, Germany) were also used. Children were continuously monitored with a pediatric pulse oximetry system and an infrared camera. All children remained asleep during the MRI scan, and no motion artifacts were present in the analyzed data.

[0451] After image alignment, non-brain signal removal, and correction of the main magnetic field and transmit magnetic field (B0 and B1) inhomogeneities, the three-pool signal model (including myelin-associated water; intra-extra-axonal water; and non-exchangeable free pool) was fitted to the mcDESPOT data to derive voxel-based MWF maps.

[0452] Then, the maps of each child were non-linearly aligned to a study-specific template. White matter masks corresponding to 5 bilateral regions (frontal, temporal, occipital, parietal, and cerebellar WM) and the body, genu, and splenium of the corpus callosum were generated from a common database, registered to the common template, and overlaid on the MWF maps of each child. Then, the mean value of each region was determined for each child and used for subsequent developmental analysis and trajectory modeling.

[0453] Developmental differences :

[0454] To examine the developmental differences between breastfed and formula-fed infants and between infants fed different formulas, a non-linear mixed effects modeling approach was used. The modified Gompertz growth model was independently fitted to Group 1 and Group 2, and to each formula subgroup. Then, an unpaired t-test was used to compare each of the four free Gompertz model parameters between the breastfed and formula-fed groups, and analysis of variance followed by a post hoc Tuckey test was used to compare each of the four free Gompertz model parameters between the four formula subgroups to determine which formula groups differed.

[0455] Cognitive assessment and analysis

[0456] In addition to MR imaging, the Mullen Scales of Early Learning {MSEL; Mullen: 1995vd} were used to evaluate each child's general cognitive abilities and skills within 7 days of scanning. The MSEL provides a broad assessment of behavioral development in fine and gross motor control, receptive and expressive language, and visual reception. Age-standardized T scores from these areas can be combined into three composite scores: the Early Learning Composite (ELC, including fine motor, visual reception, expressive and receptive language); the Nonverbal Developmental Quotient (NVDQ, containing fine motor and visual reception scores); and the Verbal Developmental Quotient (VDQ, including expressive and receptive language scores).

[0457] As with the MWF MRI data, potential group mean differences in ELC, VDQ, and NVDQ were examined between breastfed and formula-fed infants and between different formula subgroups. In addition to mean comparisons, mixed effects modeling (assuming linear trends) was used to study the longitudinal changes in these three composite values.

[0458] Example 1

[0459] Identification of nutrition-driven factors by cross-sectional analysis

[0460] From the above groups, children up to 5 years of age who were fed different infant formulas during infancy were included in a large correlation analysis to examine the relationship between the nutritional composition of the formula and brain myelination. The nutritional composition of the 5 most commonly used formulas in this group was analyzed. A single general linear model (GLM) was constructed to model all quantified nutrients, sphingomyelin, and child age. Voxel-based correlations were calculated, with significance defined as p < 0.05, and a cluster-based correction method was used to correct for type I errors.

[0461] In the initial analysis, including all nutrients resulted in an underpowered model. To reduce the number of variables in the model, we examined the correlations between variables. Using a conservative threshold of 0.9, we excluded variables (nutritional components) that were highly correlated with each other in the various formulas.

[0462] The final model showed that phospholipids (especially sphingomyelin, phosphatidylcholine, and / or phosphatidylinositol) are nutritional drivers of myelination. Other identified drivers include fatty acid derivatives (especially ARA and / or DHA), minerals (especially iron, zinc, calcium, phosphorus, magnesium), folic acid, vitamin B12, and choline.

[0463] For phospholipids, such as sphingomyelin, a significant correlation with myelination (myelin water fraction) was observed over time in the brain (especially in the cerebellum, internal capsule, visual cortex, and frontal lobe, etc.).

[0464] Example 2

[0465] Myelination trajectory according to longitudinal study

[0466] Longitudinal developmental trajectory analyses were performed on two formula foods containing different amounts of the identified nutritional driver, sphingomyelin (the composition of these formula foods is reported in Table 2 below):

[0467] Table 2 :

[0468]

[0469] The results are reported in Figure 1 .

[0470] The figure shows that the higher the sphingomyelin content in the formula food, the closer the myelin formation trajectory of the infants fed with this formula food is to the trajectory provided by exclusive human breast milk feeding.

[0471] Example 3

[0472] Measurement of cognitive ability

[0473] Cognitive abilities (Mullen Early Learning Scales) were measured in infants fed with HBM, formula food 4 (low / without sphingomyelin), and formula food 5 (high sphingomyelin).

[0474] The results are reported in Table 3 below, expressed as mean (standard deviation)

[0475] Table 3 :

[0476]

[0477]

[0478] Comparing breast milk feeding with formula food 5, there were no statistically significant differences. However, there were significant differences between breast milk feeding and formula food 4 in the expressive and receptive language scores and visual reception scores of the Mullen Early Learning Scales. Comparing formula food 5 with formula food 4, there were also those statistically significant differences in visual reception and expressive language.

[0479] Based on the results reported in the experimental section, it has been demonstrated that a nutritional composition comprising phospholipids, their metabolic precursors and / or their metabolites or mixtures thereof (such as sphingomyelin and / or phosphatidylcholine and / or phosphatidylinositol) determines the best myelin formation trajectory in the brain in infants fed with such a composition (close to the trajectory determined by human breast feeding). It has also been demonstrated with data related to cognitive abilities that this optimal trajectory in brain myelin formation corresponds to an effective improvement in the better cognitive abilities of infants, matching the abilities of breast-fed infants and balancing the lack of sub-optimal cognitive abilities shown in infants fed with formula foods low in phospholipids or free of detectable levels of phospholipids (such as sphingomyelin and / or phosphatidylcholine and / or phosphatidylinositol).

[0480] Example 3

[0481] Co-culture of neurons and OLs

[0482] Neurons / oligodendrocytes were cultured as previously described by Charles et al., 2000.

[0483] Pregnant female rats (Wistar rats) at 17 days of gestation were sacrificed by cervical dislocation and the fetuses were removed from the uterus. The forebrains were removed and placed in ice-cold Leibovitz medium (L15) containing 2% penicillin-streptomycin (PS) and 1% bovine serum albumin (BSA). The forebrains were dissociated by trypsinization for 20 minutes at 37 °C (trypsin EDTA 1X). The reaction was terminated by adding Dulbecco's modified Eagle's medium (DMEM) containing DNase I grade II (0.1 mg / ml) and 10% fetal calf serum (FCS). The cells were then mechanically dissociated by aspirating and extruding 3 times through a 10 ml pipette. The cells were then centrifuged at 180 × g for 10 minutes on a layer of BSA (3.5%) in L15 medium at 4 °C. The supernatant was discarded and the pelleted cells were resuspended in DMEM containing 10% FCS. The cells were then centrifuged at 515 × g for 10 minutes at 4 °C. The supernatant was discarded and the pelleted cells were resuspended in a medium consisting of a basal medium supplemented with 2% B27, 2 mM L-glutamine (L Glu), 2% PS solution, 1% FCS and 10 ng / ml platelet-derived growth factor (PDGF-AA). Viable cells were counted in a Neubauer cell counter using the trypan blue exclusion test. The cells were seeded at a density of 20,000 cells / well in 96-well plates pre-coated with poly-L-lysine and laminin.

[0484] One day after seeding (day 1 of culture), the cells were treated with the test compound ( Selected from those listed in Table 3) incubated with estradiol. Control cells were not incubated with the test compound or estradiol. Estradiol was used as a positive control. It is known that estradiol can induce OPC proliferation. The positive effect of estradiol on OL differentiation has also been confirmed, and it also has an impact on the early myelination process. The positive effect of estradiol on neurite growth has also been reported (for a review, see Alevaro et al., 2010).

[0485] In a humidified incubator, the plates were maintained at 37 °C in an atmosphere of air (95%) - CO2 (5%). Half of the medium was replaced every other day with fresh medium and the test compound or the control compound. The test compound or the control compound was maintained at a defined concentration during the experiment. The test compound was tested on 1 culture (6 wells per condition). Then, at days 12, 18, or 30 of culture, the cells were used to measure OPC proliferation, the differentiation of OPCs into OLs, and the early myelination process (myelin wrapping), or OL maturation (myelin maturation) and the mature myelination process (myelin wrapping).

[0486] Measurement of OPC proliferation - A2B5 positive cells and total axon length (NF)

[0487] At day 12 of culture, the cells were fixed with a cold mixture of absolute ethanol (95%) and glacial acetic acid (5%) for 5 minutes. Then the cells were permeabilized and non-specific sites were blocked with a phosphate-buffered saline (PBS) solution containing 0.1% saponin and 1% FCS at room temperature for 15 minutes.

[0488] Then the cells were incubated with monoclonal anti-A2B5 conjugated to alexa 488 (1 / 200 dilution in PBS containing 1% FCS, 0.1% saponin) at room temperature for 2 hours and with anti-NF (Neurofilament 200, phosphorylated and non-phosphorylated) raised in rabbits (1 / 500 dilution in PBS containing 1% FCS, 0.1% saponin) at room temperature for 2 hours. The antibody was developed with Alexa Fluor 568 goat anti-rabbit (1 / 400 dilution in PBS containing 1% FCS, 0.1% saponin) at room temperature for 1 hour.

[0489] The total number of OPCs (number of A2B5-positive cells) was quantified (to evaluate proliferation), and the axonal network (total axon length (NF)) was measured to assess the effect of the compound on the neuronal network (the quality of myelination is directly related to the quality of the axonal network).

[0490] Differentiation of OPCs into OLs and myelination process (myelin wrapping) - Measurement of the number and area of MAG positive cells, overlap of MAG / NF wrapping, and total axon length (NF) Maturation of OLs (myelin maturation) - Measurement of the number and area of MBP positive cells, overlap of MBP / NF wrapping, and total axon length (NF)

[0491] On the 18th day of culture, the cells were fixed with a cold mixture of absolute ethanol (95%) and glacial acetic acid (5%) for 5 minutes. Then the cells were permeabilized and non-specific sites were blocked with a phosphate-buffered saline (PBS) solution containing 0.1% saponin and 1% FCS at room temperature for 15 minutes.

[0492] The cells were then incubated with monoclonal anti-MAG produced in mice (1 / 400 diluted in PBS containing 1% FCS, 0.1% saponin), and with anti-NF (Neurofilament 200, phosphorylated and non-phosphorylated) produced in rabbits (1 / 500 diluted in PBS containing 1% FCS, 0.1% saponin) at room temperature for 2 hours. The cells were developed with CF 488A goat anti-mouse (1 / 800 diluted in PBS containing 1% FCS, 0.1% saponin) and Alexa Fluor 568 goat anti-rabbit (1 / 800 diluted in PBS containing 1% FCS, 0.1% saponin) at room temperature for 1 hour.

[0493] Quantify the total number of OLs (number and area of MAG-positive cells) (to evaluate the differentiation process), and the OPC wrapping around axons (overlapping MAG / NF wrapping) (myelination process). Measure the axonal network (total axon length (NF)) to evaluate the effect of the compound on the neuronal network.

[0494] Total axon length (NF) Table 3

[0495] On the 30th day of culture, the cells were fixed with a cold mixture of absolute ethanol (95%) and glacial acetic acid (5%) for 5 minutes. Then the cells were permeabilized and non-specific sites were blocked with a phosphate-buffered saline (PBS) solution containing 0.1% saponin and 1% FCS at room temperature for 15 minutes.

[0496] The cells were then incubated with monoclonal anti-MBP produced in mice (1 / 1000 diluted in PBS containing 1% FCS, 0.1% saponin), and with anti-NF (Neurofilament 200, phosphorylated and non-phosphorylated) produced in rabbits (1 / 500 diluted in PBS containing 1% FCS, 0.1% saponin) at room temperature for 2 hours. The cells were developed with CF 488A goat anti-mouse (1 / 800 diluted in PBS containing 1% FCS, 0.1% saponin) and Alexa Fluor 568 goat anti-rabbit (1 / 400 diluted in PBS containing 1% FCS, 0.1% saponin) at room temperature for 1 hour.

[0497] Evaluate the total number of OLs (number and area of MBP-positive cells) (to assess OL maturation) and myelin wrapping around axons (overlap MBP / NF (wrapping)). Measure the axonal network (total axon length (NF)) to assess the effect of the compound on the neuronal network.

[0498] For all measurements, once the culture is complete (6 wells per condition). For each test condition, 30 images (each image representing a field of view) were acquired and analyzed per well using an ImageXpress (Molecular devices) with 20x magnification and equipped with LED lights (excitation wavelengths 360 / 480 / 565 and emission wavelengths 460 / 535 / 620). 30 photos were automatically acquired, representing 80% of the total surface of the culture well.

[0499] Results are expressed as the cumulative average length (in μm) of the neurite network or myelin labeled with a given marker (MAG or MBP) per field of view. The overlapping area between NF and MAG or MBP was measured to evaluate wrapping.

[0500] To evaluate the OPC population, MAG-positive cell population, and MBP-positive cell population, the number of positive cells per image (= field of view) was automatically counted. Results are expressed as the average number of positive cells per field of view.

[0501] All images were acquired under the same conditions.

[0502] Figures 4 to 24

[0503]

[0504]

[0505] Results are shown in Example 4 in.

[0506] Materials and methods

[0507] 1. Feeder layer preparation: Dissociation of neonatal cortex and maintenance of mixed glial cell cultures

[0508] 2. Preparation of hippocampal neurons

[0509] Add freshly dissected brains to a 37 °C water bath for 3 minutes, then cut the cortex into small pieces using the tip of a P1000 pipette. Add 75 μL of OPC papain solution to each brain, then incubate the tissue in a 37 °C water bath for 20 minutes. Then add a mixed glial cell culture to the tissue suspension to inactivate the OPC papain solution.

[0510] Subsequently, the tissues were ground using a sterile flame-polished glass Pasteur pipette, and then 4 mL of mixed glial cell medium was added to each brain. The cells were centrifuged at 1200 rpm (about 300 g) for 5 minutes, and then the cells were resuspended in warm mixed glial cell medium and plated in a PLL-coated flask.

[0511] Four hours after plating, a complete medium change was performed to remove most of the debris caused by grinding and to promote culture viability. After 3 days of culture, a 2 / 3 medium change was performed, and no subsequent medium changes were made. Then the cells were maintained in culture until confluence.

[0512] 3. Purification of OPCs from mixed glial cell cultures to establish OL / hippocampal neuron co-cultures

[0513] Hippocampal neurons were isolated from the embryos (E18) of Sprague Dawley rats. Briefly, after the animals were sacrificed, the brains were isolated, the meninges were removed from the medial surface of the cerebral hemispheres, and then the hippocampi were dissected out and kept at 4 °C until the process was completed.

[0514] Then the tissues were incubated with 2.5% trypsin in a 37 °C water bath for 15 minutes, then gently washed and stored in the medium. The hippocampi were dissociated by repeatedly aspirating them with a functionalized sterile Pasteur pipette. After mechanical dissociation, the cells were plated at the desired density in neuronal plating medium, allowed to recover for 4 hours, and then placed in complete neuronal medium.

[0515] Figure 46

[0516] On the 9th day of mixed glial cell culture, the flasks were shaken on an orbital shaker at 50 rpm for 45 minutes in a 5% CO2 tissue culture incubator. The purpose of this shaking was to remove any loosely adherent contaminating cells from the monolayer.

[0517] Then the medium was changed, and 4 mL of fresh mixed glial cell medium supplemented with 5 μg / mL insulin was used instead. Then the flasks were placed back on the shaker and equilibrated for about 3 hours, and then shaken at 220 rpm for about 160 hours (overnight).

[0518] The next morning, the mixed glial cell medium containing microglial cells and OPC cells was collected and pre-plated in a P100 culture dish (untreated with culture) for 30 minutes to purify the OPC cells; the microglial cells immediately began to adhere to the dish, while the OPC cells remained in the supernatant medium.

[0519] Thirty minutes after pre-plating, the medium was collected, the OLs were counted and plated on hippocampal neurons in OL medium with a final volume of 1 mL.

[0520] Perform a complete replacement of the OL medium (without CNTF), and then maintain the cells in culture until the appropriate experimental time.

[0521] For the maturation experiment, the experimental procedure is as follows, as Figure 47 shown:

[0522] a. Culture OPCs on an astrocyte feeder layer in vitro for 10 days

[0523] b. Isolate OPCs (day 0)

[0524] c. Administer the compound (day 3)

[0525] d. Quantitatively evaluate maturation on days 4, 7, and 10.

[0526] For the myelination experiment, the experimental procedure is as follows, as 4. Image acquisition shown:

[0527] a. Culture hippocampal neurons until the neural network is fully mature (14 days in vitro)

[0528] b. Concurrently culture OPCs on an astrocyte feeder layer in vitro for 10 days

[0529] c. Isolate OPCs and co-culture with neurons (day 14)

[0530] d. Administer the compound (day 15)

[0531] e. Quantitatively evaluate myelination on day 15 of culture (1 day after co-culture plating and before compound treatment), days 18, 21 / 23, and 28 / 29.

[0532] OPC papain solution (made in MEM)

[0533] All cultures at different experimental time points were fixed in 4% paraformaldehyde and 4% sucrose for 10 minutes at room temperature (RT). The primary and secondary antibodies were applied in GDB buffer (30 mM phosphate buffer, pH 7.4, containing 0.2% gelatin, 0.5% Triton X-100, and 0.8 M NaCl) for 2 hours at room temperature. The cells were stained with appropriate markers (primary antibodies used: anti-A2B5 antibody (ABCAM, catalog number ab53521), rat anti-MBP (BIO-RAD, catalog number aa82-87), oligodendrocyte marker O4 antibody (R&D Systems, catalog number MAB1326), anti-βIII tubulin mAb (Promega, catalog number G7121); secondary antibodies used: Alexa anti-rat 555 (Life Tech A-21434), Alexa anti-mouse 488 (LifeTech A-11009)). After immunocytochemical staining, all images were obtained using Array Scan XTI (ThermoScientific); the objective lens was 20x, and the 2x2 pixel binning mode was adopted. For each condition and replicate well (in triplicate), at least 15 images were acquired.

[0534] To analyze all the acquired images, HCS Studio cell analysis software, specifically the "Scan" application, was used.

[0535] Mixed glial cell medium (made in DMEM)

[0536] Papain solution 1.54 mg / mL

[0537] L-cysteine 360 μg / mL

[0538] DNase I 60 μg / mL

[0539] OL medium

[0540] FBS 10%

[0541] Pen / Strep (0.33%, from stock solution) 33 units / mL of penicillin and 33 μg / mL of streptomycin

[0542] Glutamine 1%

[0543] Figures 26 to 45

[0544] DMEM

[0545] 100X OL-supplement

[0546] Bovine insulin (from 1 mg / mL stock solution)

[0547] Glutamine

[0548] Transferrin holoferric (from 33 mg / mL stock solution)

[0549] B27 supplement

[0550] FBS

[0551] CNTF (from 50 ng / μL stock solution)

[0552] The results are shown in ​ below.

Claims

1. A nutritional composition for infants and / or toddlers, the nutritional composition comprising phospholipids, their metabolic precursors and / or their metabolites, for promoting and / or supporting an optimal myelination trajectory in the brain.

2. The composition according to claim 1, wherein the optimal myelination trajectory in the brain is close to the myelination trajectory observed in infants breastfed only with human milk in the early months after birth.

3. The composition according to any one of claims 1 or 2, wherein the composition promotes and / or supports a healthy state characterized by optimal brain and cognitive function development and / or prevents neurocognitive deficits.

4. The composition according to any one of claims 1 to 3, the composition comprising an amount of sphingomyelin and / or phosphatidylcholine and / or phosphatidylinositol higher than 200 mg / Kg.

5. The composition according to any one of claims 1 to 4, the composition further comprising iron.

6. The composition according to any one of claims 1 to 5, the composition further comprising DHA.

7. The composition according to any one of claims 1 to 6, the composition further comprising folic acid.

8. The composition according to any one of claims 1 to 7, wherein the composition further comprises choline.

9. Use of the composition according to any one of claims 1 to 8 for promoting and / or supporting an optimal myelination trajectory in the brain of an infant or toddler.

10. Use of the composition according to any one of claims 1 to 8 for preventing a sub-optimal myelination trajectory in the brain of an infant or toddler.

11. A method for promoting and / or supporting an optimal myelination trajectory in the brain of an infant and / or toddler, the method comprising administering to the infant and / or toddler a nutritional composition comprising sphingomyelin, its metabolic precursors and / or its metabolites.

12. The method according to claim 11, wherein the composition according to any one of claims 1 to 8 is administered to the infant and / or toddler.

13. A nutritional composition, the nutritional composition comprising: An amount of sphingomyelin and / or phosphatidylinositol and / or phosphatidylcholine higher than 200 mg / kg; An amount of iron higher than 5 mg / 100 g; An amount of choline higher than 30 mg / 100 g; An amount of DHA higher than 30 mg / 100 g; An amount of folic acid higher than 50 mcg / 100 g.

14. A nutritional composition, the nutritional composition comprising: An amount of sphingomyelin and / or phosphatidylinositol and / or phosphatidylcholine in the range of 200 mg / kg to 2.5 g / kg; An amount of iron in the range of 5 mg / 100 g to 40 mg / 100 g; An amount of choline in the range of 30 mg / 100 g to 1000 mg / 100 g; An amount of DHA in the range of 30 mg / 100 g to 300 mg / 100 g; An amount of folic acid in the range of 50 mcg / 100 g to 500 mcg / 100 g.

Images