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. In particular, the present invention relates to a nutritional composition comprising a vitamin or a mixture thereof for 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] The present application is a divisional application, the parent application being an application with the application date of December 13, 2016, the application number of 201680072966.3, and the invention name of “Nutritional composition and infant formula 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, the present invention relates to nutritional compositions comprising vitamins or mixtures thereof for promoting or supporting an optimal myelination trajectory in the brain that approximates that observed in infants fed exclusively 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 provides a suitable alternative to natural feeding using human breast milk. Nutritional compositions for infants and young children are usually sold as powders to be reconstituted with water or in some cases as ready-to-drink or concentrated liquid compositions. These compositions are intended 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 nutrition for infants. Infant formula manufacturers have made many attempts to produce nutritional health effects that approach or resemble the benefits of human breast milk.

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

[0006] Yet in many instances, studies have shown that infant formula does not have the same effects on the body as human breast milk.

[0007] For example, infants fed infant formula and infants fed human breast milk (HBM) can exhibit different trajectories of myelination.Myelination normally occurs in the brain beginning prenatally and continues postnatally until early adulthood.

[0008] Myelination is a fundamental process of neural development that involves sheathing of axons with a fatty myelin sheath that facilitates action potential transmission. Myelination has been suggested to play a key role in coordinated communication between brain cells and networks. Myelinated white matter matures with cognitive and learning abilities.

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

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

[0011] "Thus, the results of this study suggest that not only is it important that myelination occurs to avoid cognitive deficits, but that it also is important that myelination occurs early in an infant's life, ideally following the trajectory (myelination over time) that occurs in breastfed infants. Furthermore, the study suggests that standard infant formulas do not typically provide this optimal trajectory of myelination."

[0012]

[0006] Therefore, there is a need to provide a nutritional composition for an infant or young child that will promote and / or support an optimal myelination trajectory in the brain that approximates that observed in infants fed exclusively on human breast milk (HBM).

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

[0014] There is also a need for infants fed infant formula to be provided with optimal nutrition that can promote and / or support an optimal myelination trajectory in the brain that approximates that observed in infants fed exclusively on human breast milk (HBM).

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

[0016] There is a need to compensate for the suboptimal myelination observed in non-breastfed infants. Summary of the invention

[0017] The present invention relates to a nutritional composition for infants and young children, such as a preterm formula, an infant formula, an follow-on formula, a growing-up milk or a baby food, preferably an infant formula. The composition comprises vitamins or a mixture thereof. The vitamins or a mixture thereof 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, a nutritional composition for infants and / or young children is provided, comprising vitamins or mixtures thereof for promoting and / or supporting an optimal myelination trajectory in the brain that approximates that observed in infants fed exclusively with human breast milk (HBM).

[0019] In one aspect, a nutritional composition for infants and / or young children is provided, comprising vitamins or mixtures thereof for promoting and / or supporting an optimal myelination trajectory in the brain, which trajectory approximates that observed in infants fed exclusively with human breast milk (HBM), which trajectory 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 vitamins or mixtures thereof for the manufacture of 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 trajectory approximates that observed in infants fed exclusively with human breast milk (HBM).

[0021] In another aspect, there is provided the use of vitamins or mixtures thereof for the manufacture of 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 trajectory approximates that observed in infants fed exclusively with human breast milk (HBM), which trajectory 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, a method is provided for promoting and / or supporting an optimal myelination trajectory in the brain that approximates that observed in infants fed exclusively with human breast milk (HBM), the method comprising administering to the infant and / or young child a nutritional composition comprising vitamins or a mixture thereof.

[0023] In another aspect, a method is provided for promoting and / or supporting an optimal myelination trajectory in the brain, the method comprising administering to an infant and / or young child a nutritional composition comprising vitamins or a mixture thereof, which trajectory approximates that observed in infants fed exclusively with human breast milk (HBM) and results in promoting and / or supporting a health state characterized by optimal brain and cognitive function development and / or preventing neurocognitive deficits.

[0024] In another aspect, there is provided the use of vitamins or mixtures thereof for promoting and / or supporting an optimal myelination trajectory in the brain in infants and / or young children, which trajectory approximates that observed in infants fed exclusively with human breast milk (HBM).

[0025] In another aspect, there is provided the use of vitamins or mixtures thereof for promoting and / or supporting an optimal myelination trajectory in the brain in infants and / or young children that approximates that observed in infants fed exclusively with human breast milk (HBM) and that results in a health state characterized by promoting and / or supporting optimal brain and cognitive function development and / or preventing neurocognitive deficits.

[0026] The infant or toddler may be between 0 and 60 months old, or between 0 and 24 months old, or between 0 and 12 months old, or between 0 and 6 months old.

[0027] In another aspect of the present invention, there is provided a nutritional composition, which, in addition to vitamins (e.g., folic acid and / or vitamin B12) or mixtures thereof, further comprises:

[0028] Minerals (particularly iron, magnesium, phosphorus, copper, calcium and / or zinc), phospholipids (e.g. sphingomyelin), phospholipids, their metabolic precursors and / or their metabolites, fatty acid derivatives (e.g. DHA and / or ARA, nervonic acid and / or stearic acid) and / or choline.

[0029] In another aspect of the present invention, a nutritional composition comprising sphingomyelin, iron, choline, DHA and folic acid is provided. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 : Shows the myelination trajectory of infants and toddlers fed with breast milk compared to infants and toddlers fed with two commercial formulas containing different levels of folic acid. Data were obtained from the experiments described in Example 2.

[0031] Figure 2 : The structure of 2-amino-4-octadecene-1,3-diol (sphingosine defined below) is shown.

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

[0033] Figure 4 : The effect of DHA on MBP, NF and / or MBP / NF at day 18 and / or day 30 is shown.

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

[0035] Figure 6 : The effects of B12 on A2B5, MBP / NF, NF and MAG at day 12, day 30 and / or day 18 are shown.

[0036] Figure 7 : The effect of vitamin B12 on A2B5, NF, MBP / NF and / or MAG at day 12, day 18 and / or day 30 is shown.

[0037] Figure 8 : The effects of folic acid on A2B5, NF, MAG, MAG / NF and / or MBP / NF at day 12, day 18 and / or day 30 are shown.

[0038] Fig. 9 : The effects of choline on A2B5, MAG and / or MBP at day 12, day 18 or day 30 are shown.

[0039] Fig.10 : The effect of zinc on MBP, NF and / or MBP / NF at day 12, day 18 and / or day 30 is shown.

[0040] Fig.11 : The effect of phosphorus on MAG, NF and / or MAG / NF at day 12, day 18 and / or day 30 is shown.

[0041] Fig.12 : The effect of magnesium on A2B5, MBP, NF, MAG, MBP / NF and / or MAG / NF at day 12, day 18 and / or day 30 is shown.

[0042] Fig.13 : The effect of copper on A2BF, MAG and / or MAG / NF at day 12 and / or day 18 is shown.

[0043] Fig.14 : Shown are the effects of phosphatidylcholine on A2B5 at day 12 and on MAG at day 18.

[0044] Fig.15 : The effects of phosphatidylinositol on A2B5, MBP, MAG, NF, MAG / NF at day 12, day 18 and / or day 30 are shown.

[0045] Fig.16 : The effect of phosphatidylserine on A2B5, NF and / or MAG / NF at day 12 and / or day 18 is shown.

[0046] Fig.17 : The effect of sphingomyelin on A2B5, MAG and / or MBP at day 12, day 18 and / or day 30 is shown.

[0047] Fig.18 : Shown are the effects of ceramide on A2B5 at day 12 and on MAG at day 18.

[0048] Fig.19 : The effects of galactosylceramide on A2B5, MBP, NF and / or MBP / NF at day 12 and / or day 30 are shown.

[0049] Fig. 20 : The effect of glucosylceramide on A2B5 at day 12 and on NF at days 12 and 18 are shown.

[0050] Fig.21 : Shown are the effects of D-erythroceramide on A2B5 at day 12 and on MAG at day 18.

[0051] Fig. 22 : Shown are the effects of ceramide-1-phosphate on A2B5 at day 12 and on NF and MAG at day 18.

[0052] Fig.23 : The effects of monosialoganglioside-3 (GM3) on A2B5, MBP, MAG and / or MBP / NF at day 12, day 18 and / or day 30 are shown.

[0053] Fig.24 : The effects of disialoganglioside 3 (GD3) on A2B5, MBP, NF and / or MAG at day 12, day 18 and / or day 30 are shown.

[0054] Fig.25 : The fatty acid profiles of phosphatidylinositol (PI), phosphatidylcholine (PC), phosphatidylserine (PS) and sphingomyelin used in Example 3 are shown.

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

[0056] Fig. 27 : Shown are the effects of ARA on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

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

[0058] Fig.29 : Shown are the effects of zinc on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

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

[0060] Fig.31 : Shown are the effects of GD3 on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0061] Fig.32 : Shown are the effects of DHA on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0062] Fig.33 : Shown are the effects of neuraminic acid on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0063] Fig.34 : Shown are the effects of iron on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

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

[0065] Fig.36 : Shown are the effects of phosphatidylserine on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

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

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

[0068] Fig.39 : Shown are the effects of ceramide on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0069] Fig.40: Shown are the effects of galactosylceramide on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

[0070] Fig.41 : Shown are the effects of glucosylceramide on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

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

[0072] Fig.43 : Shows the effect of D-erythroceramide on MAG and MBP mRNA expression and on the co-expression of MBP and BetaIII.

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

[0074] Fig.45 : Shows the effect of GM3 on the co-expression of MBP and BetaIII. DETAILED DESCRIPTION

[0075] definition

[0076] As used herein, the following terms have the following meanings.

[0077] In the context of the present invention, the term "trajectory of myelination" refers to the extent of myelination (related to myelin water fraction) over time during infancy and early childhood.

[0078] In the context of the present invention, the term "optimal trajectory of myelination" means a myelination trajectory as defined above which is as close as possible to the myelination trajectory achieved in infants who are exclusively breastfed during the first (3) months of life.

[0079] If the distance between the myelination trajectory of an infant and any equivalent / identical measurement point on the myelination trajectory of an infant exclusively breastfed during the first (3) months of life is up to 50%, particularly up to 25%, more particularly up to 20%, then the myelination trajectory of the infant may be considered as close as possible to the myelination trajectory achieved in the exclusively breastfed infant. Non-limiting examples within the range of 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 bioequivalent.

[0080] 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 person's life, more particularly, within the first 2 and first 3 years of a person's life, and even more particularly, within the first year of a person's life.

[0081] The myelin formation trajectory can be determined by measuring the myelin associated water fraction and / or myelin associated water pool in the subject at different time points, particularly in the first 5 years of human subject life, more particularly in the first 2 years and first 3 years of human life, and even more particularly in the first year of human life. The myelin formation trajectory can be determined by measuring the myelin associated water fraction and / or myelin associated water pool in the subject using multi-component relaxation (MCR) magnetic resonance imaging (MRI) technology and particularly using the mcDESPOT technology (Deoni et al. 2008). In particular, the myelin formation trajectory can be determined by measuring the myelin associated water pool using the mcDESPOT technology (Magn. Reson. Med. 2008 60: 1372-1387, the subject matter of which is hereby incorporated herein by reference).

[0082] In the context of the present invention, the term "facilitates" refers to a factor or factors that cause a process to occur.

[0083] In the context of the present invention, the term "support" refers to a factor or factors that maintain a process once it has started to occur.

[0084] In the context of the present invention, the expression "learning" refers to the acquisition of knowledge or skills through experience, study or through teaching.

[0085] The term "cognitive" refers to the intellectual processes by which an individual knows, perceives, or understands ideas; thus, the ability to think and understand. Cognition includes all aspects of information processing, perception, attention, thinking, reasoning, comprehension, and memory, as well as psychomotor, language, memory, concentration, executive function, and problem-solving abilities.

[0086] The term "infant" refers to a child under the age of 12 months.

[0087] The expression "infant" means a child between the ages of one and five years old (including toddlers).

[0088] The expression "child" generally refers to a human being up to the age of eighteen years.

[0089] A "preterm baby" or "preterm infant" is a baby or toddler born before the due date. It usually refers to a baby born before 37 weeks of gestation.

[0090] The expression "full-term infant" refers to an infant born after 37 weeks of gestation.

[0091] In the context of the present invention, the term "low birth weight infant" refers to a newborn baby having a weight of less than 2500 g (5.5 lbs) due to preterm birth (ie before 37 weeks of gestation) and / or due to fetal growth restriction.

[0092] 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 for infants of the same gestational age.

[0093] The expression "postpartum period" is the period starting immediately after the birth of a child and lasting about six weeks.

[0094] The expression "nutritional composition" refers to a composition that provides nutrients to a subject. Generally, the nutritional composition is ingested enterally or parenterally, or orally or intravenously, and generally comprises a lipid or fat source and a protein source. Preferably, the nutritional composition is for oral use.

[0095] The expression "hypoallergenic nutritional composition" refers to a nutritional composition which is less likely to cause an allergic reaction.

[0096] The expression "synthetic composition" refers to a mixture obtained by chemical and / or biological methods, the chemical properties of which may be identical to the mixture naturally occurring in mammalian milk.

[0097] The expression "infant formula" means a food intended for the nutrition of infants from 4 to 6 months of age and which meets the multiple nutritional needs of such infants (as provided for in Article 1.2 of Council Directive 91 / 321 / EEC of 14 May 1991 on infant formula and follow-on formula).

[0098] The expression "stage 1 infant formula" refers to food specifically used to provide nutrition to infants under 4 months of age.

[0099] The term "follow-on formula" refers to a food which is designed to provide nutrition to infants over 4 months of age and is the principal liquid food in the progressively more diversified diet of such infants.

[0100] In the context of the present invention, the term "growing-up milk (GUM)" means a nutritional formula that can be given to children after infant formula has been stopped. "Growing-up milk" (or GUM) is used from the age of one year. It is usually a milk-based beverage adapted to the special nutritional needs of young children.

[0101] The term "baby food" refers to food specially designed to provide nutrition for infants under 1 year old.

[0102] The expression "fortifier" refers to a liquid or solid nutritional composition suitable for mixing with breast milk or infant formula.

[0103] The term "weaning" refers to the period during which other foods are gradually replacing the mother's milk in the infant's diet.

[0104] “Mother's milk” is understood to mean mother's breast milk or colostrum (=human breast milk=HBM).

[0105] The term "oligofructose" used herein refers to fructose oligomers. It can be long chain or short chain, depending on the degree of polymerization (number of monomers) of the oligofructose. Preferably, the oligofructose of the present invention is a short-chain oligofructose, most preferably it has a degree of polymerization of 2 to 10, for example a degree of polymerization of 2 to 8.

[0106] As used herein, the term "sn-2 palmitate" refers to palmitic acid bonded to a triglyceride at the sn-2 position.

[0107] "High sn-2 palmitic acid triglyceride" refers to a triglyceride (TG) containing more than 30% palmitic acid at the sn-2 position. For example, a commercially available high sn-2 palmitate ingredient 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.

[0108] "Alpha-lactalbumin" refers to a high-quality, easily digestible whey protein that accounts for 20%-25% of the total human breast milk (HBM) protein and is the main protein present in HBM. The structure of alpha-lactalbumin consists of 123 amino acids and 4 disulfide bonds, and the protein has a molecular weight of 14.2K Daltons. Since alpha-lactalbumin has a high content of essential amino acids, in particular tryptophan, it is an ideal choice for low-protein infant formulas. According to the present invention, alpha-lactalbumin also represents a source of sphingomyelin.

[0109] The term "prebiotic" refers to non-digestible carbohydrates that have a beneficial effect on the host by selectively stimulating the growth and / or activity of healthy bacteria, such as bifidobacteria in the human colon (Gibson GR, Roberfroid MB. Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. J Nutr. 1995; 125: 1401-12).

[0110] The term "probiotic" refers to a microbial cell preparation or microbial cell component that has 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: 10107-10 (Salminen S., Ouwehand A., Benno Y. et al., "How to define probiotics", "Trends in Food Science and Technology", 1999, Vol. 10, pp. 107-110)). Microbial cells are generally bacteria or yeast.

[0111] The term "sphingosine" means a Figure 2 2-amino-4-octadecene-1,3-diol of the structure. As described below, sphingosine can constitute the backbone of sphingolipids.

[0112] The term "sphingoid base" refers to sphingosine and a class of compounds structurally derived therefrom, such as dihydrosphingosine, dihydrosphingosine and phytosphingosine. The term "sphingoid base" also includes phosphorylated forms of dihydrosphingosine, sphingosine, dihydrosphingosine and phytosphingosine. In one embodiment, the sphingoid base is selected from: dihydrosphingosine, sphingosine and phosphorylated forms thereof.

[0113] The term "sphingolipids" refers to a class of lipids containing a sphingoid base backbone in which the amino group (-NH2) is acylated by a fatty acid residue. "Ceramide" and "sphingomyelin" as defined below are exemplary sphingolipids.

[0114] 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 may refer to a single ceramide substance as well as a mixture of single ceramide substances. In particular, the ceramide is a compound of formula (IXa) or a mixture of compounds of formula (IXa)

[0115]

[0116] in,

[0117] R16a is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group,

[0118] R17a is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group.

[0119] 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.

[0120] Non-limiting examples of C14 to C44 saturated or unsaturated fatty acids from which the fatty acid residue can 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. :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.

[0121] 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 chosen 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, chosen from C16:0, C18:0, C20:0, C22:0 and C24:0.

[0122] 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 together with the adjacent carbonyl groups) contained in the mixture is predominantly saturated fatty acids and least predominantly unsaturated fatty acids. More particularly, the mixture is such that 80% to 96% of the fatty acid residues in the mixture are saturated fatty acids, particularly C14, C15, C16, C18, C20, C22, C23, C24 saturated fatty acids, more particularly C16, C18, C20, C22 and C24.

[0123] The term "ganglioside" as used herein refers to an oligosaccharide ceramide lipid molecule comprising a residue of ceramide of formula IXa as defined herein. When the term ganglioside is used in this specification, it may refer to a single ganglioside species as well as a mixture of single ganglioside species comprising a residue of ceramide of formula IXa as defined herein.

[0124] The term "sphingomyelin" refers to a lipid molecule in which the sphingosine backbone is acylated at the amino group (-NH2) by a fatty acid residue, and in which the hydroxyl group at position 1 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 substance and a mixture of single sphingomyelin substances, wherein the preferred fatty acid residue is a residue of a C14 to C44 fatty acid, non-limiting examples of which include C14 to C44 saturated or unsaturated fatty acids, and non-limiting examples of which C14 to C44 saturated or unsaturated fatty acids from which the fatty acid residues 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.

[0125] The term "choline" refers to a compound containing an N,N,N-trimethylethanolammonium cation and having Figure 3 Quaternary ammonium salts of the structures reported in.

[0126] In the context of the present invention, the term "choline" shall be intended to refer to all choline present in the nutritional composition of the present invention, either in free form (or in the form of its salts) or in a form derived from a structure comprising 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 composition of the present invention, either in free form or in the form of its salts (such as choline hydroxide).

[0127] The term "fatty acid derivative" as used herein refers to compounds containing fatty acids other than phospholipids, and in particular refers to free fatty acids, and / or monoacylglycerol (hereinafter referred to as MAG) and / or diacylglycerol (hereinafter referred to as DAG), and / or triacylglycerol (hereinafter referred to as TAG) and / or cholesterol ester. More particularly, the term refers to MAG, DAG, TAG and / or cholesterol ester. Even more particularly, the term refers to TAG.

[0128] The term "MAG" as used herein 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)

[0129]

[0130] in,

[0131] R 18 , R 19 or R 20 Two of them are H, and R 18 , R 19 or R 20 One of the groups is a C4 to C44 saturated or unsaturated acyl group.

[0132] More specifically, R 18 , R 19 or R 20 Two of them are H and R 18 , R 19 or R 20 One of the is a C10 to C24 saturated or unsaturated acyl group, and even more particularly a C14 to C24 saturated or unsaturated acyl group.

[0133] 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)

[0134] in,

[0135] R 18 , R 19 or R 20 One of them is H, and R 18 , R 19 or R 20 Two of them are C4 to C44 saturated or unsaturated acyl groups. More particularly, they are C10 to C24 saturated or unsaturated acyl groups, and even more particularly, they are C14 to C24 saturated or unsaturated acyl groups. The two C4 to C44 saturated or unsaturated acyl groups may be the same or different.

[0136] 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)

[0137] in,

[0138] Where R 18 , R 19 or R 20All 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 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] in,

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

[0143] More specifically, 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] in

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

[0148] More specifically, R 22 is a C9 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group, and even more particularly a C13 to C23 branched or unbranched acyclic alkyl or acyclic alkenyl group.

[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" shall be intended to refer to all DHA present in the nutritional composition of the present invention, either in free form (as fatty acids or physiologically acceptable salts thereof) or in a form contained in a fatty acid derivative structure.

[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" should be intended to refer to all ARA present in the nutritional composition of the present invention, either in free form (as fatty acid or its physiologically acceptable salt) or in a form contained in a fatty acid derivative structure.

[0151] The term "vitamin" as used herein 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 mean all folic acid present in the nutritional composition of the present invention, either as such or in the form of one of its physiologically acceptable salts (folate) 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 usually 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, either 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 ferric EDTA) and mixtures thereof.

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

[0156]

[0157] in,

[0158] R 1 is O;

[0159] X is NH or O;

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

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

[0162]

[0163] Among them, R 5 is a C2-C44 saturated or unsaturated, linear or branched acyl group, and

[0164] R 6is a C2-C44 saturated alkyl or alkenyl group; and

[0165] R 4 is selected from: C5 or C6 substituted or unsubstituted cycloalkyl or cycloalkenyl groups, or

[0166] —(CH2)n—R 7 , wherein n is an integer in the range of 1 to 4, in particular 1 to 2, and R 7 is -N(CH3)3+, NH3+ or a substituent of formula (IV), and

[0167]

[0168] Especially R 4 is a C6 cycloalkyl or alkyl or alkenyl group substituted by one or more hydroxyl groups, more particularly R 4 Derived from inositol (C6H12O6), and even more particularly myo-inositol, i.e. R 4 for:

[0169]

[0170] As used herein, the term "acyclic" refers to a group that is not cyclic, ie, does not contain a closed chain of atoms.

[0171] In the context of the present invention, the term "phosphatidylinositol" means a compound of formula (V)

[0172]

[0173] Where R 8 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group, and

[0174] R 9 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group.

[0175] More specifically, R 8 and R 9 are independently C13 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl groups which together with their adjacent carbonyl groups correspond to a C14 to C44 saturated or unsaturated fatty acid residue, even more particularly, R 8 and R 9 Independently of one another 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.

[0176] More specifically, R 8 and R 9is a C13 to C23 branched or unbranched acyclic alkyl or acyclic alkenyl group, which together with their adjacent carbonyl groups is 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, 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.

[0177] In the context of the present invention, the term "phosphatidylserine" means a compound of formula (VI)

[0178]

[0179] Where R 10 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group, and

[0180] R 11 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group.

[0181] More specifically, R 10 and R 11 are independently C13 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl groups which together with their adjacent carbonyl groups correspond to a C14 to C44 saturated or unsaturated fatty acid residue, even more particularly, R 10 and R 11 Independently of one another 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.

[0182] More specifically, R 10 and R 11 is a C13 to C23 branched or unbranched acyclic alkyl or acyclic alkenyl group, which together with their adjacent carbonyl groups is 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, 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.

[0183] In the context of the present invention, the term "phosphatidylethanolamine" means a compound of formula (VII)

[0184]

[0185] Where R 12 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group, and

[0186] R 13 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group.

[0187] More specifically, R 12 and R 13 are independently C13 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl groups which together with their adjacent carbonyl groups correspond to a C14 to C44 saturated or unsaturated fatty acid residue, even more particularly, R 12 and R 13 Independently of one another 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.

[0188] In the context of the present invention, the term "phosphatidylcholine" means a compound of formula (IX)

[0189]

[0190] Where R 16 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group, and

[0191] R 17 is a C2 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl group.

[0192] More specifically, R 16 and R 17 are independently C13 to C43 branched or unbranched acyclic alkyl or acyclic alkenyl groups which together with their adjacent carbonyl groups correspond to a C14 to C44 saturated or unsaturated fatty acid residue, even more particularly, R 16 and R 17 Independently of one another 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.

[0193] More specifically, R 16 and R 17is a C13 to C23 branched or unbranched acyclic alkyl or acyclic alkenyl group, which together with their adjacent carbonyl groups is 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, 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.

[0194] The term "cfu" is to be understood as colony forming units.

[0195] Exclusively breastfed infant or toddler / infant or toddler fed solely with breast milk: has the usual meaning and refers to an infant whose majority of nutrients and / or energy comes from human breast milk ("majority" may be at least 90% or at least 95%, or at least 99%).

[0196] Infants / young children fed primarily with infant formula: has the usual meaning, meaning that the nutritional source of nutrients and / or energy is derived primarily from synthetic infant formula, follow-on milk or growing-up milk. "Primarily" means at least 50% of those nutrients and / or energy, or at least 75%.

[0197] Unless otherwise indicated, all percentages are by weight.

[0198] Now the invention will be described in more detail. It should be noted that the various aspects, features, embodiments and implementations described in this application may be compatible and / or may be combined together.

[0199] In addition, in the context of the present invention, the term "comprising" or "including" does not exclude other possible elements. The composition mentioned in the present invention (including the multiple embodiments described herein) may include the following elements, be composed of or consist essentially of the following elements: the basic elements and necessary restrictions described herein, and any other or optional (or depending on the needs) ingredients, components or restrictions described herein.

[0200] The terms "particularly" or "more particularly" as used herein should not be considered limiting, but should be interpreted as being synonymous with "for example" or "especially".

[0202] For the sake of clarity, all embodiments and aspects reported hereinafter shall apply mutatis mutandis to the different embodiments and aspects described for the present invention.

[0203] Unless otherwise indicated, all amounts indicated for nutrients are expressed as amounts per dry weight of the nutritional composition.

[0204] Where a nutrient may be contained in the composition in different forms (either as such or in the form of a salt, complex or more complex structure comprising the nutrient), the amounts reported below should be intended to refer to the amount of the nutrient itself.

[0205] The nutritional composition according to the invention comprises vitamins or mixtures thereof.

[0206] Particularly effective vitamins may be folic acid, vitamin B12 and vitamin B6, especially folic acid and vitamin B12, especially folic acid.

[0207] Vitamins may be included in the compositions of the invention in amounts ranging from 0.001% up to 99.999% of the composition.

[0208] In one embodiment, the composition of the invention comprises vitamin B12 and / or folic acid.

[0209] Folic acid may be included in the composition of the invention in an amount of 0.001% up to 99.999% of the composition.

[0210] In particular, folic acid may be included in an amount higher than 50 mcg / 100 g of the dry composition, more particularly from 50 to 500 mcg / 100 g of the dry composition.

[0211] Besides minerals or mixtures thereof, such as iron, zinc, calcium, phosphorus and / or magnesium, the nutritional composition according to the invention also comprises folic acid.

[0212] Folic acid may be included in the composition of the invention in an amount of 0.001% up to 99.999% of the composition.

[0213] The nutritional composition according to the invention comprises folic acid.

[0214] In one embodiment, the nutritional composition according to the invention comprises folic acid in an amount higher than 50 mcg / 100 g.

[0215] In one embodiment, the nutritional composition according to the invention comprises folic acid in an amount higher than 110 mcg / 100 g.

[0216] In one embodiment, the nutritional composition according to the invention comprises folic acid in an amount ranging from 50 to 500 mcg / 100 g or from 50 to 400 mcg / 100 g.

[0217] In another embodiment, the nutritional composition according to the invention comprises folic acid in an amount ranging from 110 to 500 mcg / 100 g or from 110 to 400 mcg / 100 g.

[0218] In another embodiment, the nutritional composition according to the invention comprises folic acid in an amount ranging from 110 to 400 mcg / 100 g or from 110 to 350 mcg / 100 g.

[0219] In one embodiment, the nutritional composition according to the invention contains folic acid at a level such that the total daily intake from the nutritional composition of the invention will not exceed 400 mcg.

[0220] Folic acid may be incorporated into the nutritional composition of the invention as such or in the form of one of its physiologically acceptable salts (folate) or a mixture thereof.

[0221] In another embodiment, the nutritional composition according to the invention further comprises vitamin B12.

[0222] Vitamin B12 may be included in the compositions of the invention in an amount of 0.001% up to 99.999% of the composition.

[0223] In particular, vitamin B12 may be contained 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, wherein all weights are per 100 g dry composition.

[0224] In one embodiment, the composition of the present invention comprises vitamin B12 in an amount selected from: above 0.5mcg, ranging from 0.1mcg to 10mcg, 0.4mcg to 5mcg, 0.5mcg to 2mcg, 1mcg to 1.5mcg, 4mcg to 8.5mcg, wherein all weights are per 100g dry composition.

[0225] In one embodiment, the composition according to the invention comprises an amount of vitamin B12 such that the total daily intake from the nutritional composition of the invention will not exceed 7.6 mcg / 100 g dry composition (77.6 mcg / Kg dry composition).

[0226] Vitamin B12 may be incorporated into the nutritional composition of the present invention as such or in the form of its physiologically acceptable salt or mixture thereof or via any source comprising vitamin B12. In particular, vitamin B12 may be incorporated into the composition in its pure form, as cyanocobalamin, hydroxocobalamin and any combination thereof.

[0227] In one embodiment, the nutritional composition according to the invention comprises, in addition to its mineral mixture, for example in addition to the vitamins, one or more of the following ingredients: minerals and / or phospholipids and / or fatty acids and / or choline.

[0228] It may be particularly advantageous if the composition according to the invention further comprises one or more of the following ingredients: minerals and / or phospholipids and / or fatty acid derivatives and / or choline.

[0229] In one embodiment, the nutritional composition according to the invention comprises, in addition to the vitamins or mixtures thereof, phospholipids, metabolic precursors thereof and / or metabolites thereof (eg sphingomyelin, phosphatidylcholine and / or phosphatidylinositol).

[0230] The phospholipids, their metabolic precursors and / or their metabolites may be contained in the composition in an amount of up to 99.999% of the composition.

[0231] In particular, sphingomyelin, its metabolic precursors and / or its metabolites may be contained in the composition in an amount of up to 99.999% of the composition.

[0232] More particularly, the composition will comprise sphingomyelin in an amount higher than 200 mg / kg dry weight of the composition, more particularly ranging from 200 mg to 2.5 g / kg dry weight of the composition.

[0233] In one embodiment, the composition comprises sphingomyelin in an amount selected from: above 200 mg / kg, above 300 mg / kg, in a range of 200 mg / kg to 2.5 g / kg, in a range of 200 mg / kg to 2 g / kg, in a 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.

[0234] In one embodiment, phosphatidylcholine, its metabolic precursor and / or its metabolite may be included in the composition in an amount of up to 99.999% of the composition.

[0235] More particularly, the composition will comprise phosphatidylcholine in an amount higher than 200 mg / kg dry weight of the composition, more particularly ranging from 200 mg / kg to 2.5 g / kg dry weight of the composition.

[0236] In one embodiment, the composition comprises phosphatidylcholine in an amount selected from: greater than 200 mg / kg, greater than 300 mg / kg, in a range of 200 mg / kg to 2.5 g / kg, in a range of 200 mg / kg to 2 g / kg, in a 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.

[0237] In particular, phosphatidylinositol, its metabolic precursors and / or its metabolites may be contained in the composition in an amount of up to 99.999% of the composition.

[0238] More particularly, the composition will comprise phosphatidylinositol in an amount higher than 200 mg / kg dry weight of the composition, more particularly ranging from 200 mg / kg to 1.5 g / kg dry weight of the composition.

[0239] In one embodiment, the composition comprises phosphatidylinositol in an amount selected from: greater than 200 mg / kg, greater than 300 mg / kg, in a range of 200 mg / kg to 2.5 g / kg, in a range of 200 mg / kg to 2 g / kg, in a 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.

[0240] In particular, phosphatidylserine, its metabolic precursors and / or its metabolites may be contained in the composition in an amount of up to 99.999% of the composition.

[0241] More particularly, the composition will comprise phosphatidylserine in an amount higher than 150 mg / kg dry weight of the composition, more particularly ranging from 200 mg / kg to 1.5 g / kg dry weight of the composition.

[0242] In one embodiment, the composition comprises phosphatidylserine in an amount selected from: greater than 150 mg / kg, greater than 200 mg / kg, greater than 300 mg / kg, an amount ranging from 200 mg / kg to 2.5 g / kg, an amount ranging from 200 mg / kg to 2 g / kg, an amount ranging from 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.

[0243] In particular, phosphatidylethanolamine, its metabolic precursor and / or its metabolite may be contained in the composition in an amount of up to 99.999% of the composition.

[0244] More particularly, the composition will comprise phosphatidylethanolamine in an amount higher than 150 mg / kg dry weight of the composition, more particularly ranging from 150 mg / kg to 1.5 g / kg dry weight of the composition.

[0245] In one embodiment, the composition comprises phosphatidylethanolamine in an amount selected from: greater than 170 mg / kg, greater than 180 mg / kg, greater than 200 mg / kg, an amount ranging from 200 mg / kg to 2.5 g / kg, an amount ranging from 200 mg / kg to 2 g / kg, an amount ranging from 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.

[0246] 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.

[0247] 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 compound may be used in an amount such that the level of phospholipids physiologically delivered by the composition is consistent with the levels listed above. It is within the capabilities of the skilled artisan to determine the appropriate amount.

[0248] As used herein, the term "metabolic precursors and / or metabolites of one or more phospholipids" does not include choline.

[0249] Non-limiting examples of metabolic precursors and / or metabolites of phospholipids, in particular sphingomyelin, phosphatidylcholine, phosphatidylinositol, phosphatidylserine and / or phosphatidylethanolamine, are: galactosylceramide, glucosylceramide, sphingosine, sphingosine-1-phosphate, ceramide, D-erythro-dihydroceramide and ceramide-1-phosphate and gangliosides.

[0250] Particularly effective phospholipids may be phosphatidylcholine, phosphatidylserine, phosphatidylinositol and / or sphingomyelin, in particular sphingomyelin.

[0251] 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 substances and / or a combination of any of the foregoing substances. In particular, the phospholipid is sphingomyelin, its metabolic precursor and / or its metabolite.

[0252] Particularly effective metabolic precursors and / or metabolites of phospholipids, in particular sphingomyelin, include ceramides and gangliosides as well as ceramide-1-phosphate and d-erythro-dihydroceramide.

[0253] Particularly effective gangliosides may be monosialoganglioside-3 (GM3) ganglioside and / or disialoganglioside 3 (GD3) ganglioside.

[0254] Ceramide-1 -phosphate and d-erythro-dihydroceramide will comprise the residue of ceramide of formula IXa as defined herein.

[0255] Gangliosides and / or ceramides and / or ceramide-1-phosphate and / or d-erythro-dihydroceramide may be included in the composition in any amount.

[0256] Concentrations in the range of 2 mg / 100 g to 11.5 mg / 100 g of GD3 and / or GM3 may be particularly effective.

[0257] Sphingomyelin can be synthesized from ceramide and / or one or more gangliosides and phosphatidylcholine, and therefore it may be particularly advantageous if ceramide and / or one or more gangliosides are used in combination with phosphatidylcholine, its metabolic precursors or its metabolites.

[0258] The phospholipids, its metabolic precursors and / or its metabolites included in the composition of the present invention can be natural, synthetic or their mixture. The metabolic precursors and / or metabolites can be used in the composition of the present invention in its pure form or substantially pure form. As another option, they can be added in the form of a source that comprises them.

[0259] Any source of phospholipids, metabolic precursors thereof, and / or metabolites thereof suitable for ingestion by a subject intended to consume the composition may be used in the present invention.

[0260] In particular, phospholipids, their metabolic precursors or their metabolites will come from natural sources, non-limiting examples of which include eggs, soybeans, bovine brains and / or mammalian milk or extracts thereof. Non-limiting examples of soy sources include soy lecithin-food additives, and non-limiting examples of mammalian milk include cow's milk, camel milk, sheep milk, goat milk, including skim milk. Non-limiting extracts of milk include protein extracts (such as whey proteins and caseins), milk fat globule membranes (MFGM) and extracts containing them.

[0261] Particularly useful sources of phospholipids, metabolic precursors or metabolites thereof (particularly sphingomyelin) useful in the present invention may be milk whey protein concentrate enriched with α-lactalbumin, and / or non-purified α-lactalbumin that has been extracted from whey protein (particularly milk whey protein).

[0262] Alpha-lactalbumin is a high quality, easily digestible whey protein and is the major protein present in human milk. Alpha-lactalbumin and / or milk fractions enriched in alpha-lactalbumin are ideal for use in low-protein infant formulas due to their high content of essential amino acids, particularly tryptophan. Although alpha-lactalbumin itself is a protein, impure sources may contain sphingomyelin.

[0263] In one embodiment, the phospholipids, their metabolic precursors or their metabolites, in particular sphingomyelin, are used in the form of a whey protein concentrate enriched in α-lactalbumin or as α-lactalbumin.

[0264] In a more specific embodiment, an α-lactalbumin-enriched whey protein concentrate or α-lactalbumin is used, having a phospholipid content, in particular a sphingomyelin content, higher than 500 mg / 100 g dry weight of the composition.

[0265] Another particularly useful source of phospholipids, their metabolic precursors or their metabolites may be MFGM or an extract containing them, in particular MFGM from milk or an extract containing them. It may be particularly advantageous if the MFGM or an extract containing them contains 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, in particular in a concentration range of 0.05% to 2%, 0.1% to 0.4%.

[0266] Those skilled in the art can determine the appropriate amount of the above-mentioned nutrients, their metabolic precursors or their metabolites based on the nature, purpose, target subject and dosage of the composition, such as how many times a subject is to ingest the composition per day. Typically, the effective dose will depend on the age, size and health of the subject, the lifestyle of the subject, the amount of the nutrients in the composition, and possibly the sex of the subject.

[0267] In one embodiment, the nutritional composition according to the invention comprises, in addition to vitamins (eg folic acid or vitamin B12) or mixtures thereof, sphingomyelin.

[0268] In one embodiment, the nutritional composition according to the invention comprises sphingomyelin in an amount higher than 200 mg / kg.

[0269] In another embodiment, the nutritional composition of the present invention comprises sphingomyelin in an amount higher than 300 mg / kg.

[0270] 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.

[0271] In another embodiment, the nutritional composition according to the invention comprises sphingomyelin in an amount ranging from 200 to 2 g / kg, such as from 300 mg / kg to 1.5 g / kg or from 400 mg / kg to 1 g / Kg.

[0272] In the context of the present invention, sphingomyelin can be administered as is 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 cause the same beneficial effects in promoting and / or supporting the optimal myelination trajectory in the brain or preventing the myelination trajectory from being optimal in the brain compared to the direct administration of sphingomyelin dosage. Non-limiting exemplary substances in this regard are: galactosylceramide, glucosylceramide, sphingosine, sphingosine-1-phosphate, ceramide, D-erythro-dihydroceramide and ceramide-1-phosphate and gangliosides.

[0273] The nutritional composition of the present invention comprises sphingomyelin and / or its metabolic precursor or metabolite.

[0274] Sphingomyelin is present in natural sources such as α-lactalbumin, eggs, bovine brain or milk.

[0275] Sphingomyelin and / or its metabolic precursors or metabolites thereof may be incorporated into the nutritional composition of the present invention as a single substance, as an ingredient consisting of a mixture of different sphingomyelin substances or by adding natural or synthetic ingredients containing one or more sphingomyelin substances.

[0276] In one embodiment, sphingomyelin may be incorporated into the nutritional composition of the present invention in the form of being included in skimmed milk powder, α-lactalbumin, whey protein concentrate and / or whey protein concentrate enriched with α-lactalbumin. In one embodiment, the whey protein concentrate enriched with α-lactalbumin may have a sphingomyelin content of more than 500 mg / 100 g.

[0277] In one embodiment, sphingomyelin may be incorporated into the nutritional composition of the present invention as a single ingredient.

[0278] In one embodiment, the nutritional composition according to the invention contains metabolic precursors and / or metabolites of sphingomyelin in an amount such that they physiologically deliver the same level of sphingomyelin as delivered by a nutritional composition comprising 200 mg / kg to 2.5 g / kg of sphingomyelin.

[0279] In another embodiment, the nutritional composition according to the invention comprises metabolic precursors and / or metabolites of sphingomyelin in an amount such that they physiologically deliver the same level of sphingomyelin as delivered by a nutritional composition comprising 400 mg / Kg to 1.5 g / Kg of sphingomyelin.

[0280] The skilled person will arrive at such amount based on his knowledge in this field.

[0281] In one embodiment, the nutritional composition according to the invention comprises a mineral or a mixture thereof.

[0282] Particularly effective minerals may be iron, zinc, calcium, phosphorus, copper and magnesium, especially iron.

[0283] In one embodiment, the composition of the invention comprises iron and / or zinc and / or calcium and / or phosphorus and / or copper and / or magnesium, in particular iron and zinc, more in particular iron.

[0284] In another embodiment, the nutritional composition according to the invention comprises, in addition to folic acid, iron.

[0285] In one embodiment, the nutritional composition according to the invention comprises iron in an amount higher than 5 mg / 100 g.

[0286] In one embodiment, the nutritional composition according to the invention comprises iron in an amount higher than 9 mg / 100 g.

[0287] In one embodiment, the nutritional composition according to the invention comprises iron in an amount ranging from 5 mg / 100 g to 40 mg / 100 g or from 9 mg / 100 g to 40 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 to 20 mg / 100 g or from 9 mg / 100 g to 20 mg / 100 g.

[0289] In another embodiment, the nutritional composition according to the invention comprises iron in an amount ranging from 5 mg / 100 g to 15 mg / 100 g or from 9 mg / 100 g to 15 mg / 100 g.

[0290] In addition to folic acid, the nutritional composition of the present invention may further comprise iron.

[0291] Iron may be incorporated into the nutritional composition of the present invention 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.

[0292] Iron may 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 a mixture thereof.

[0293] In one embodiment, the nutritional composition according to the invention comprises iron at a level such that the total daily intake from the nutritional composition of the invention will not exceed 40 mg.

[0294] In another embodiment, the nutritional composition according to the invention comprises zinc.

[0295] Zinc may be included in the compositions of the invention in an amount of 0.001% up to 99.999% of the composition.

[0296] In particular, zinc may be included in the composition in an amount higher than 0.08 mg, higher than 0.3 mg, higher than 0.5 mg, wherein all weights are per 100 g dry composition.

[0297] In one embodiment, the composition according to the invention comprises zinc in an amount selected from the range 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, wherein all weights are per 100 g dry composition.

[0298] 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.

[0299] Zinc may be incorporated into the composition of the invention in the form of a physiologically acceptable salt such as, for example, zinc nitrate, zinc sulfate, zinc gluconate, zinc acetate or mixtures thereof, or in the form of a physiologically acceptable zinc complex such as, for example, zinc picolinate or mixtures thereof.

[0300] In another embodiment, the nutritional composition according to the invention comprises calcium.

[0301] Calcium may be included in the compositions of the invention in an amount from 0.001% up to 99.999% of the composition.

[0302] In particular, calcium may be included in the composition in an amount higher than 0.84 mg, higher than 2.52 mg, higher than 4.62 mg, wherein all weights are based on 100 g of dry weight of the composition.

[0303] In one embodiment, the composition according to the invention comprises calcium in an amount ranging from 84 mg to 760 mg, ranging from 200 mg to 550 mg, ranging from 250 mg to 450 mg, ranging from 280 mg to 520 mg, 350 mg to 650 mg, wherein all weights are per 100 g dry composition.

[0304] In one embodiment, the composition according to the invention comprises calcium at a level such that the total daily intake from the nutritional composition of the invention will not exceed 482 mg / day, or will not exceed 477 mg / day.

[0305] Calcium 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 comprising calcium, for example calcium carbonate, calcium chloride, calcium salts of citric acid, calcium gluconate, calcium glycerophosphate, calcium lactate, calcium hydroxide, calcium salts of orthophosphoric acid.

[0306] In another embodiment, the nutritional composition according to the invention comprises magnesium.

[0307] Magnesium may be included in the compositions of the present invention in an amount of 0.001% up to 99.999% of the composition.

[0308] In particular, magnesium may be included in the composition in an amount higher than 0.2 mg, higher than 0.35 mg, higher than 0.5 mg, wherein all weights are based on 100 g of dry weight of the composition.

[0309] In one embodiment, the composition according to the invention comprises magnesium in an amount selected from the range of 0.35 mg to 90 mg, ranging from 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, wherein all weights are per 100 g dry composition.

[0310] In one embodiment, the composition according to the invention comprises magnesium at a level such that the total daily intake from the nutritional composition of the invention will not exceed 110 mg / day, or will not exceed 65 mg / day.

[0311] 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 comprising magnesium, for example magnesium carbonate, magnesium chloride, magnesium oxide, magnesium sulfate, magnesium gluconate, magnesium hydroxide, magnesium salts of citric acid, magnesium salts of orthophosphoric acid.

[0312] In another embodiment, the nutritional composition according to the invention comprises phosphorus.

[0313] Phosphorus may be included in the compositions of the present invention in an amount of 0.001% up to 99.999% of the composition.

[0314] In particular, phosphorus may be contained in the composition in an amount higher than 1.7 mg / 100 g of dry weight of the composition, higher than 14.3 mg / 100 g of dry weight of the composition, higher than 27.3 mg / 100 g of dry weight of the composition.

[0315] In one embodiment, the composition according to the invention comprises phosphorus in an amount selected from the range of 17 mg to 516 mg, range of 129 mg to 400 mg, range of 140 mg to 390 mg, range of 150 mg to 370 mg, 160 mg to 365 mg, range of 270 mg to 350 mg, 200 mg to 360 mg, wherein all weights are per 100 g dry composition.

[0316] In one embodiment, the composition according to the invention comprises phosphorus at a level such that the total daily intake from the nutritional composition of the invention will not exceed 863 mg / day, or will not exceed 787 mg / day.

[0317] Phosphorus may be incorporated into the composition of the invention as such or in the form of its physiologically acceptable salts and / or via any source comprising phosphorus, such as: calcium phosphate, calcium hydrogen phosphate.

[0318] In another embodiment, the nutritional composition according to the invention comprises copper.

[0319] Copper may be included in the compositions of the present invention in amounts ranging from 0.001% up to 99.999% of the composition.

[0320] In particular, copper may be included in the composition in an amount greater than 10 meg, greater than 40 meg, greater than 60 meg, wherein all weights are based on 100 g of dry weight of the composition.

[0321] In one embodiment, the composition according to the invention comprises copper in an amount chosen from: higher than 100mcg, ranging from 100mcg to 850mcg, 180mcg to 650mcg, 200mcg to 400mcg, 210mcg to 300mcg, 210mcg to 240mcg, 450mcg to 850mcg, wherein all weights are based on dry weight per 100g of the composition.

[0322] In one embodiment, the composition according to the invention comprises copper at a level such that the total daily intake from the nutritional composition of the invention will not exceed 1426 mcg / day, or will not exceed 488 mcg / day.

[0323] Copper 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 comprising 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.

[0324] In addition to vitamins (such as folic acid and / or vitamin B12) or mixtures thereof, the nutritional composition of the present invention may further comprise choline and / or metabolic precursors or metabolites thereof.

[0325] In another embodiment, the nutritional composition according to the invention comprises, in addition to folic acid, choline.

[0326] In one embodiment, the nutritional composition according to the invention comprises choline in an amount higher than 30 mg / 100 g.

[0327] 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.

[0328] In one embodiment, the nutritional composition according to the invention comprises choline in an amount ranging from 30 mg / 100 g to 1000 mg / 100 g or from 30 mg / 100 g to 700 mg / 100 g.

[0329] In another embodiment, the nutritional composition according to the invention comprises choline in an amount ranging from 50 mg / 100 g to 1000 mg / 100 g or from 50 mg / 100 g to 700 mg / 100 g.

[0330] In another embodiment, the nutritional composition according to the invention comprises choline in an amount ranging from 50 mg / 100 g to 500 mg / 100 g or from 100 mg / 100 g to 400 mg / 100 g.

[0331] In one embodiment, the nutritional composition according to the invention comprises choline at a level such that the total daily intake from the nutritional composition of the invention will not exceed 1 g.

[0332] 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 a mixture 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.

[0333] Choline and / or its metabolic precursors or metabolites thereof may be incorporated into the nutritional composition of the present invention as a single substance, as an ingredient consisting of a mixture of different choline substances or by adding natural or synthetic ingredients containing one or more choline substances.

[0334] In addition to folic acid, the nutritional composition of the present invention may further comprise choline and / or its metabolic precursors or metabolites thereof.

[0335] In one embodiment, the nutritional composition according to the invention further comprises fatty acid derivatives or mixtures thereof.

[0336] Non-limiting examples of C10 to C44 saturated or unsaturated fatty acids that may be contained in the fatty acid derivatives, i.e., fatty acids that may be free fatty acids or fatty acid residues from which MAG, DAG, TAG and / or cholesterol esters may be derived, 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, C20:3n-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, and in particular C24:2n-6. 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, C4 0:0, C41:2, C41:1, C41:0, C42:47, C42:3, C42:2, C42:1, C42:0, C44:3, C44:1. In particular, the fatty acid 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, in particular 24:1n-9.

[0337] It may be particularly advantageous if the fatty acid derivative comprises 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 in particular 22:6n-3 and / or C18:0.

[0338] Any fatty acid derivative suitable for ingestion by a subject intended to consume the composition may be used in the present invention.

[0339] In particular, fatty acid derivatives will come from natural sources, non-limiting examples of which include eggs, algae, fish oil, molds, yeasts, seeds, plants such as soybeans, and animal sources such as bovine brains, and / or mammalian milk or extracts thereof. Non-limiting examples of soybean sources include soy lecithin-food additives, 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, milk fat globule membranes (MFGM) and extracts containing them. Fatty acid derivatives can also come from palm oil, tallow, lard, cottonseed oil, peanut oil.

[0340] It may be particularly advantageous if the fatty acid derivative comprises 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 in particular 22:6n-3 and / or C18:0.

[0341] 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.

[0342] In one embodiment, the composition according to the invention contains fatty acid derivatives comprising DHA and / or ARA and / or nervonic acid and / or stearic acid, in particular fatty acid derivatives comprising DHA and / or ARA and / or stearic acid. Most in particular fatty acid derivatives comprising DHA and / or stearic acid.

[0343] Fatty acid derivatives comprising DHA and / or ARA and / or nervonic acid and / or stearic acid may be included in the composition of the present invention in an amount of up to 99.999% of the composition.

[0344] In particular, fatty acid derivatives comprising DHA and / or ARA and / or nervonic acid and / or stearic acid may be included in the composition of the present 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.

[0345] In one embodiment, the composition according to the present invention contains fatty acid derivatives 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 of the composition and 300 mg / 100 g dry weight of the composition, in the range of 30 mg / 100 g dry weight of the composition to 200 mg / 100 g dry weight of the composition, or in the range of 30 mg / 100 g dry weight of the composition to 150 mg / 100 g dry weight of the composition, in the range of 50 mg / 100 g dry weight of the composition to 300 mg / 100 g dry weight of the composition, in the range of 50 mg / 100 g dry weight of the composition to 200 mg / 100 g dry weight of the composition, in the range of 50 mg / 100 g dry weight of the composition to 150 mg / 100 g dry weight of the composition.

[0346] Fatty acid derivatives including stearic acid occur in natural sources such as palm oil, tallow, lard, cottonseed oil, peanut oil.

[0347] 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.

[0348] Fatty acid derivatives comprising DHA and / or ARA are present in natural sources such as, for example, eggs, algae, fungi or fish oils.

[0349] The oil containing the fatty acid derivative that comprises DHA and / or ARA and other polyunsaturated fatty acids (PUFA) (particularly EPA (eicosapentaenoic acid)) usually can have various sources.Preferably, the fatty acid derivative that comprises DHA is provided in the form of fish oil containing the fatty acid derivative that comprises DHA and / or ARA.Fish oil usually contains 5 % by weight or more, preferably 10 % by weight or more of the fatty acid derivative that comprises DHA and / or ARA.The oil containing a large number of fatty acid derivatives that comprise DHA and / or ARA obtained from algae or microorganism is also available for use usually.For example, the oil containing 10 % by weight or more, for example 20 % by weight or more of the fatty acid derivative gathered from algae can be used.

[0350] If the nutritional composition according to the invention contains fatty acid derivatives comprising ARA and DHA, the ingredients may be included in the composition of the invention 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, in particular 1.1:1 to 1:1.1.

[0351] It may also be advantageous if the composition according to the invention comprises 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 according to 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.

[0352] In addition, when a large number of fatty acid derivatives comprising DHA and / or ARA are included in the composition of the present invention, it may be particularly advantageous if the total amount of fatty acid derivatives comprising saturated long-chain fatty acids, particularly C20 / 24, is increased. These saturated long-chain fatty acids may be important components of myelin sheaths, thereby enabling them to wrap and encapsulate axons. The weight ratio of DHA and / or AA to these unsaturated long fatty acids in the composition of the present invention may 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.

[0353] The nutritional composition according to the invention comprises ARA.

[0354] In one embodiment, the nutritional composition according to the invention comprises ARA in an amount higher than 30 mg / 100 g.

[0355] In one embodiment, the nutritional composition according to the invention comprises ARA in an amount higher than 50 mg / 100 g.

[0356] 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.

[0357] 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.

[0358] ARA is found in natural sources such as eggs, fungi, algae or fish oil.

[0359] According to one embodiment, the ARA is provided in the form of a triglyceride comprising ARA.

[0360] The oil that comprises ARA and other polyunsaturated fatty acids (PUFA) usually, particularly EPA (eicosapentaenoic acid) can have various sources.Preferably, ARA is provided in the form of the fish oil that comprises ARA.Fish oil usually comprises 5 % by weight or more, preferably 10 % by weight or more ARA.The oil that comprises a large amount of ARA obtained from algae or microorganism is also available.For example, the oil that contains 10 % by weight or more, for example 20 % by weight or more ARA gathered from fungi can be used.

[0361] ARA can be incorporated into the nutritional composition of the present 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 an ingredient consisting of a mixture of different ARA substances or by adding natural or synthetic ingredients containing one or more ARA substances.

[0362] In one embodiment, the nutritional composition of the invention comprises, in addition to ARA: phospholipids (particularly sphingomyelin), minerals (particularly iron, magnesium, phosphorus, calcium and / or zinc), choline, DHA, vitamin B12 and / or folic acid.

[0363] In another embodiment, the nutritional composition according to the invention comprises, in addition to vitamins such as folic acid and or vitamin B12, DHA.

[0364] In one embodiment, the nutritional composition according to the invention comprises DHA in an amount higher than 30 mg / 100 g.

[0365] In one embodiment, the nutritional composition according to the invention comprises DHA in an amount higher than 50 mg / 100 g.

[0366] In one embodiment, the nutritional composition according to the invention comprises DHA 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.

[0367] 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 from 50 mg / 100 g to 200 mg / 100 g or from 50 mg / 100 g to 150 mg / 100 g.

[0368] The nutritional composition of the present invention comprises DHA.

[0369] DHA is found in natural sources such as eggs, algae or fish oil.

[0370] According to one embodiment, the DHA is provided in the form of triglycerides comprising DHA.

[0371] The oil containing DHA and other polyunsaturated fatty acids (PUFA) in general, particularly EPA (eicosapentaenoic acid) can have various sources. Preferably, DHA is provided in the form of fish oil containing DHA. Fish oil generally contains 5 wt % or more, preferably 10 wt % or more of DHA. The oil containing a large amount of DHA obtained from algae or microorganisms is also available. For example, the oil containing 10 wt % or more, for example 20 wt % or more of DHA gathered from algae can be used.

[0372] 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 ingredients containing one or more DHA substances.

[0373] In one embodiment, the nutritional composition according to the invention comprises, in addition to folic acid and DHA, arachidonic acid (ARA).

[0374] According to one embodiment, the nutritional composition comprises about the same amount of DHA and ARA. For example, 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, in particular 1.1:1 to 1:1.1.

[0375] In one embodiment, when a substantial amount of DHA is included in the nutritional composition, the unsaturated long chain fatty acids c20 / 22 / 24 are increased.

[0376] In one embodiment, the nutritional composition according to the invention comprises folic acid, iron and choline.

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

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

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

[0380] In another embodiment, the nutritional composition according to the invention comprises folic acid, DHA and iron.

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

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

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

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

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

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

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

[0388] In one embodiment, the nutritional composition according to the invention comprises sphingomyelin, folic acid, iron and DHA.

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

[0390] In another embodiment, the nutritional composition according to the invention comprises sphingomyelin, folic acid, DHA and choline.

[0391] In another embodiment, the nutritional composition according to the invention comprises folic acid, iron, DHA and choline.

[0392] 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 substances; and / or minerals, in particular iron, and / or zinc, and / or calcium, and / or phosphorus, and / or magnesium; and / or choline.

[0393] Particularly beneficial concentrations / amounts of the ingredients in the composition may be sphingomyelin in an amount of at least 420 mg / kg, phosphatidylcholine in an amount of at least 1000 mg / kg, phosphatidylserine in an amount of at least 900 mg / kg, phosphatidylinositol in an amount of at least 700 mg / kg, folic acid in an amount of at least 160 mg / kg, vitamin B12 in an amount of at least 7 mcg / 100 g, iron in an amount of at least 11.5 mg / 100 g , choline in an amount of at least 140 mg / kg, fatty acid derivatives containing DHA in an amount of at least 89 mg / 100 g, fatty acid derivatives containing AA in an amount of at least 175 mg / 100 g, zinc in an amount of at least 7 mg / 100 g, calcium in an amount of at least 500 mg / 100 g, phosphorus in an amount of at least 350 mg / 100 g, copper in an amount of at least 600 mcg / 100 g, and magnesium in an amount of at least 50 mg / 100 g. All weights are based on the dry weight of the composition.

[0394] In one embodiment, the composition according to the invention comprises phospholipids and / or metabolic precursors or metabolites thereof (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.

[0395] 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.

[0396] In a more specific embodiment, the composition according to the invention contains DHA at a concentration of 1023 mg / kg, fatty acid derivatives of 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.

[0397] The composition of the present invention may be any type of composition suitable for direct administration to a subject.

[0398] In particular, the composition will be a synthetic nutritional composition.

[0399] In one embodiment, the nutritional composition according to the invention comprises sphingomyelin, folic acid, iron, choline and DHA.

[0400] One skilled in the art will determine the appropriate amounts of the above nutrients, their metabolic precursors or metabolites thereof to achieve their maximum permitted levels in the nutritional composition after administration.

[0401] Preferred nutritional composition base :

[0402] The composition according to the invention may be a synthetic nutritional composition. It may be an infant formula, a stage 1 infant formula, an follow-on formula, a growing-up milk, a baby food, a premature infant formula or a fortifier such as a human milk fortifier or supplement. Preferably, the composition of the invention is an infant formula or a fortifier or supplement specifically for infants under 4 months or under 6 months.

[0403] In a typical embodiment of the invention, the composition will contain a protein source, a lipid source, and a carbohydrate source.

[0404] For example, such a composition may comprise protein in the range of about 2 g / 100 kcal to 6 g / 100 kcal, lipid in the range of about 1.5 g / 100 kcal to 3 g / 100 kcal, and / or carbohydrate in the range of about 1.7 g / 100 kcal to 12 g / 100 kcal.

[0405] If the composition is liquid, its energy density may be between 60 kcal / 100 ml and 75 kcal / 100 ml.

[0406] If the composition is solid, its energy density may be between 60 kcal / 100 g and 75 kcal / 100 g.

[0407] The type of protein is considered to be insignificant to the present invention. Therefore, protein sources such as whey, casein and mixtures thereof can be used. As for whey protein, acid whey or sweet whey or mixtures thereof can be used, as well as alpha-lactalbumin and beta-lactoglobulin mixed in any desired proportion. The whey protein can be modified sweet whey. Sweet whey is a byproduct of the cheese making process that is more easily available and is commonly used to make infant formula based on cow's milk. However, sweet whey contains a component called casein glycomacropeptide (CGMP), which is undesirably rich in threonine and has very little tryptophan. Removing CGMP from sweet whey can make the content of threonine in the protein closer to the content of threonine in human milk. Then, those low-content amino acids (mainly histidine and tryptophan) can be supplemented in this modified sweet whey. A method for removing CGMP from sweet whey is described in EP880902, and an infant formula based on such modified sweet whey is described in WO 01 / 11990. The protein may be complete or hydrolyzed, or a mixture of complete protein and hydrolyzed protein. For example, for a subject who is considered to be at risk of milk allergy, it may be desirable to provide partially hydrolyzed protein (degree of hydrolysis between 2% and 20%). If hydrolyzed protein is required, a hydrolysis process may be performed as required and as known in the art. For example, the whey fraction may be enzymatically hydrolyzed by two steps as described in EP 322589 to prepare a whey protein hydrolyzate. For fully hydrolyzed protein, Alcalase 2.4L (EC 940459) may be used first at 55°C, then Neutrase 0.5L (available from Novo Nordisk Ferment AG), and then pancreatin may be used to triple hydrolyze whey protein. If the whey fraction used as a raw material is substantially lactose-free, it is found that the protein undergoes much less lysine blockage during the hydrolysis process. This enables the extent of lysine blocking to be reduced from about 15 wt % of total lysine to less than about 10 wt % of lysine; for example about 7 wt % of lysine, which greatly improves the nutritional quality of the protein source.

[0408] The composition of the present invention may contain a carbohydrate source. Any carbohydrate source may be used, such as lactose, sucrose, maltodextrin, starch, honey and mixtures thereof.

[0409] The composition of the present invention may contain a lipid source. The lipid source may 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, i.e. linoleic acid and alpha-linolenic acid, may also be added. In one embodiment, a small amount of oils rich in primary processed arachidonic acid (AA) and docosahexaenoic acid (DHA), such as fish oil or microbial oil, may be added. The ratio of n-6 fatty acids to n-3 fatty acids that the lipid source has is preferably from about 5:1 to about 15:1; for example, from about 8:1 to about 10:1.

[0410] Composition of the present invention also can comprise all vitamins and minerals that are reported to be indispensable and nutritionally demanded in daily diet.The minimum requirement of some vitamins and minerals has been determined.The example of minerals, vitamins and other nutrients optionally present in infant formulas comprises 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 salt.In one embodiment, said composition comprises a certain amount of vitamin B12 and / or folic acid.The existence and amount of specific minerals and other vitamins will change according to many factors, such as age, body weight and the condition of the person or animal of composition to be applied.

[0411] The composition may also include at least one probiotic strain. Probiotics are microbial cell preparations or microbial cell components 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, especially Bifidobacterium lactis CNCM I-3446 sold under the trademark Bb12 by Danish Christian Hansen company, and Bifidobacterium longum ATCC BAA-999 sold under the trademark BB536 by Japan's Morinaga Milk Industry Co., Ltd. If probiotics are present, their amount is also preferably varied according to the age of the person or animal. Generally speaking, the amount of probiotics may increase with the age of the infant, e.g. 3cfu / g formula food (dry weight) up to 10 12 cfu / g formula food (dry weight), preferably between 10 4 cfu / g formulation (dry weight) and 10 8 cfu / g formula food (dry weight).

[0412] The composition may also contain at least one prebiotic in an amount of 0.3% to 10%. Prebiotics are non-digestible food ingredients that have a beneficial effect on 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. These ingredients are non-digestible in 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 to the colon, where they are selectively fermented by beneficial bacteria. Examples of prebiotics include certain oligosaccharides.

[0413] The composition may optionally contain other substances that may have beneficial effects, such as nucleotides, nucleosides, and the like.

[0414] The composition used in the present invention, such as infant formula, can be prepared by any suitable means. For example, infant formula 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 in order to avoid thermal degradation, they are usually added later. Any lipophilic vitamins, emulsifiers and the like can be dissolved in the fat source first and then mixed. Water (preferably water that has undergone reverse osmosis) can then be mixed to form a liquid mixture. The liquid mixture can then be heat treated to reduce the amount of bacteria. For example, the liquid mixture can be rapidly heated to a temperature in the range of about 80°C to about 110°C for about 5 seconds to about 5 minutes. This can be done by steam injection or by a heat exchanger (e.g., a plate heat exchanger). The liquid mixture can then be cooled to about 60°C to about 85°C, for example, by rapid cooling. The liquid mixture can then be homogenized; for example, in two stages: the first stage is carried out at about 7MPa to about 40MPa, and the second stage is carried out at about 2MPa to about 14MPa. The homogenized mixture can then be further cooled to add any heat-sensitive components; such as vitamins and minerals. It is convenient to standardize the pH and solid content of the homogenized mixture at this moment. 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 weight %. If it is necessary to increase one or more probiotics, the probiotics can be cultured according to any suitable method, and then, for example, prepared by freeze drying or spray drying, for addition to infant formula. Alternatively, bacterial products that have been made into suitable forms can be purchased from professional suppliers such as Danish Hansen (Christian Hansen) and Japanese Morinaga Milk Industry Co., Ltd. (Morinaga) for addition to food products (such as infant formula). Such bacterial products can be added to powdered infant formula by dry mixing.

[0415] The vitamins or mixtures thereof may be added at any stage in the process but are preferably added after the heating step.

[0416] In one embodiment, the nutritional composition comprises triglycerides having high sn-2 palmitate, preferably triglycerides having more than 33% palmitic acid in the sn-2 position.

[0417] In some embodiments, palmitic acid comprises about 15% to about 25%, such as about 15% to about 20%, by weight, of the total fatty acid content of the formula, 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.

[0418] A commercially available composition sold by Lipid Nutrition Corporation is BetapolTM B-55, which is a mixture of triglycerides derived from vegetable oils in which at least 54% of the palmitic acid is located at 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% (e.g., about 43% to about 45%) Betapol TM B-55. Those skilled in the art will appreciate that the percentage of high sn-2 fat and the total amount of sn-2 palmitate used in the formula may vary and different high sn-2 palmitate oils may be used without departing from the spirit and scope of the present invention.

[0419] Health Effects

[0420] The composition of the invention has a positive effect on the myelination trajectory in the brain of an infant or young child fed with this composition.

[0421] Such positive effects may include promoting and / or supporting an optimal myelination trajectory in a subject fed with the composition, which determines the proper development of cognitive skills and abilities and learning in an infant or young child. An optimal myelination trajectory may also prevent the occurrence of cognitive impairment or delay.

[0422] The health effects may be observed after days, weeks or months of consumption of the nutritional composition comprising vitamins (eg, folic acid and / or vitamin B12).

[0423] The effects of the invention may be preventative (eg, avoiding a suboptimal myelination trajectory in the brain) or curative (restoring an optimal myelination trajectory in the brain).

[0424] The health effects associated with the infant can be measured by various methods as shown in the examples below.

[0425] Target Baby

[0426] In one embodiment of the invention, the target infant or toddler displays a suboptimal myelination trajectory in the brain, which can lead to cognitive deficits, impaired cognitive abilities, and / or suboptimal cognitive development.

[0427] In one embodiment, the infant may be a premature infant or a low birth weight infant or an infant that is small for gestational age.

[0428] In another embodiment, the infant or toddler is born full term.All infants can benefit from the present invention, as all infants are or may be susceptible to developing a suboptimal myelination trajectory in the brain.

[0429] In such infants or young children, it is particularly interesting to obtain a myelination trajectory in the brain close to that of breastfed infants, preferably infants fed exclusively with breast milk during the first few months of life. In fact, it provides them with a health status in cognitive abilities that is consistent with that observed in breastfed infants.

[0430] In one embodiment, infants and young children 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 anticipated that the compositions of the invention may be more beneficial when administered to infants just after birth (0-4 weeks, 0-8, 0-12, 0-24 weeks) because the myelination process has already begun and is significantly developed.

[0431] The following examples are presented to illustrate certain embodiments and features of the invention, but should not be construed as limiting the scope of the invention.

[0432] Anticipated feeding regimen

[0433] In one embodiment, the composition of the invention is fed to an infant or young child (or is intended to be fed or is instructed to be fed) for 2 weeks to 52 weeks. In one embodiment, it is fed to an infant or young child for 2 weeks to 24 weeks, or 2 weeks to 12 weeks.

[0434] In one embodiment, the composition of the invention is fed to an infant or young child (or is intended to be fed or is instructed to be fed) for 2 weeks to 52 weeks, and begins shortly after the infant or young child is born or breastfeeding is interrupted. In one embodiment, the composition of the invention is fed to an infant or young child for 2 weeks to 24 weeks, or 2 weeks to 12 weeks, and begins shortly after the infant or young child is born or breastfeeding is interrupted.

[0435] Without wishing to be bound by theory, it is believed that early initiation (at or shortly after birth) is preferred to induce the desired effects.

[0436] It is expected that when the composition of the present invention is used as the sole source of nutrition (in addition to supplementing breast milk feeding), the health benefits are more prominent or 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 needs (e.g., energy needs) of the target infant or young child.

[0437] Experimental Section :

[0438] Methods, definitions, and materials

[0439] MRI (Magnetic Resonance Imaging) :

[0440] MRI brain scans of infants and children between the ages of 0 and 5 years were obtained using white matter imaging techniques. This technique provides a quantitative measure, the myelin water fraction (MWF), which is a surrogate marker of myelin content. When plotted over time throughout early childhood, a myelination trajectory can be generated.

[0441] Infant formula ingredients : To analyze the composition / levels of myelin-related nutrients of six infant formulas fed to infants participating in this study.

[0442] The compositions tested were standard commercial infant formulas from different brands / suppliers and showed different levels of nutrients contained therein.

[0443] 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.

[0444] Clinical Research

[0445] Infant Participants

[0446] 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). In order to focus on typical neurodevelopment, children with known potential risk factors for learning disabilities, neurological disorders, or psychiatric disorders were explicitly excluded at recruitment and enrollment. Thus, children with in utero alcohol or illicit substance exposure, preterm birth (<37 weeks of gestation) or multiple births, fetal ultrasound abnormalities, complicated pregnancies (such as preeclampsia), APGAR scores <8, admission to the neonatal intensive care unit, neurological disorders (such as head injury, epilepsy), and psychiatric or developmental disorders 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 children from enrollment with clinically relevant behaviors or obvious medical conditions (such as autism spectrum disorders).

[0447] A combination of retrospective and prospective data was obtained from parents through a detailed medical history and interviews about the type of infant formula used, the ratio of breastfeeding to formula feeding, and the duration of exclusive breastfeeding. This information was updated at each study visit, 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 only 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 exclusive formula-fed group were further subdivided according to the main infant formula used in the first three months as reported by the parents. The main formula was defined as the formula given 90% or more of the time (e.g., when parents use an alternative brand during vacation).

[0448] Using these criteria, 94 exclusively formula-fed infants and toddlers were selected for Group 1. This included 13 children receiving Formula #2; 28 children receiving Formula #5; 8 children receiving Formula #3; 39 children receiving Formula #4; 5 children receiving Formula #1, and 1 child receiving Formula #6. A sample of 52 exclusively breastfed infants was also selected and matched to the exclusively formula-fed group with respect to mean age, gestational age, birth weight, sex ratio, ethnicity ratio, maternal education, family size, and number of languages ​​(other than English) spoken at home. Group demographics are provided in Table 1.

[0449] Table 1: Data breakdown for longitudinal and nutritional analyses

[0450]

[0451] Imaging methods and analysis

[0452] Each infant was scanned using mcDESPOT (T1 and T2 multivariate drive balanced single pulse observation technique) white matter imaging technology (Magn. Reson. Med. 2008, 60: 1372-1387) by Deoni et al., which provides quantitative measurement of myelin water fraction (MWF) at each imaging point of the whole brain. All infants were scanned using the mcDESPOT imaging protocol for anechoic during natural (i.e., non-sedated) sleep. Total imaging time ranged from 19 minutes for the youngest toddler to 24 minutes for the older 4-year-old child.

[0453] All data were collected on a Siemens 3T Tim Trio scanner equipped with a 12-channel head RF array. To minimize movement during scanning, the children were restrained with a pediatric MedVac vacuum fixation bag (CFI Medical Solutions, USA) and foam pads. Scanner noise was reduced by reducing the peak gradient amplitude and switching rate and using a soundproof scanner hole insert (Quiet Barrier HD Composite, UltraBarrier, USA). MiniMuff pediatric earmuffs and powered headphones (MR Confon, Germany) were also used. The children were constantly monitored with a pediatric pulse oximetry monitoring system and an infrared camera. All children remained asleep during the MRI scan, and no motion artifacts appeared in the analyzed data.

[0454] After image alignment, non-brain signal removal, and correction for main and emission magnetic field (B0 and B1) inhomogeneities, a three-pool signal model (including myelin-associated water; axonal intra-extracellular water; and a non-exchangeable free water reservoir) was fitted to the mcDESPOT data to derive voxel-based MWF maps.

[0455] Each child's atlas was then nonlinearly aligned to a study-specific template. Corresponding to 5 bilateral regions (frontal, temporal, occipital, parietal, and cerebellar WM) and the body, genu, and splenium of the corpus callosum

[0456] A white matter mask was generated from a common database, registered to a common template, and superimposed on the MWF atlas for each child. The mean value for each region was then determined for each child and used for subsequent developmental analysis and trajectory modeling.

[0457] Developmental differences :

[0458] To examine developmental differences between breastfed and formula-fed infants and between infants fed different formulas, a nonlinear mixed-effects modeling approach was used. A modified Gompertz growth model was independently fitted to Groups 1 and 2, and to each formula subgroup. Each of the four free Gompertz model parameters was then compared between the breastfed and formula-fed groups using an unpaired t-test, and each of the four free Gompertz model parameters was compared between the 4 formula subgroups using analysis of variance followed by a post hoc Tuckey test to determine which formula group was different.

[0459] Cognitive Assessment and Analysis

[0460] In addition to MR imaging, the general cognitive abilities and skills of each child were evaluated within 7 days of scanning using the Mullen Early Learning Scale {MSEL; Mullen:1995vd}. The MSEL provides extensive assessments of behavioral development in fine motor and gross motor control, receptive and expressive language, and visual acceptance. The age-standardized T scores from these aspects can be combined into three comprehensive scores: early learning comprehensive score (ELC, including fine motor, visual acceptance, expressive and receptive language); non-verbal development quotient (NVDQ, including fine motor and visual acceptance scores); and language development quotient (VDQ, including expressive and receptive language scores).

[0461] As with the MWF MRI data, potential group mean differences in ELC, VDQ, and NVDQ between breastfed and formula-fed infants and between the different formula subgroups were examined. In addition to mean comparisons, longitudinal changes in these three composite values ​​were investigated using mixed-effects modeling (assuming linear trends).

[0462] Example 1

[0463] Identification of nutritional drivers by cross-sectional analysis

[0464] From the above cohort, children up to 5 years of age who were fed different infant formulas during infancy were included in a grand correlation analysis to examine the relationship between formula nutritional composition and brain myelination. The nutritional composition of the 5 most commonly used formulas in this cohort was analyzed. A single general linear model (GLM) was constructed modeling all quantified nutrients, folate, and child age. Voxel-based correlations were calculated, significance was defined as p < 0.05, and type I errors were corrected using a cluster-based correction method.

[0465] In the initial analysis, the inclusion of all nutrients resulted in an underpowered model. To reduce the number of variables in the model, we examined correlations among variables. Using a conservative threshold of 0.9, we excluded variables (nutrient components) that were highly correlated with each other across formulas.

[0466] The final model suggests that vitamins (especially folic acid and vitamin B12) are nutritional drivers of myelination. Other identified drivers include fatty acid derivatives, especially ARA and / or DHA, phospholipids (especially sphingomyelin, phosphatidylcholine and / or phosphatidylinositol), minerals (especially iron, calcium, phosphorus, zinc and magnesium) and choline.

[0467] For vitamins, significant correlations with myelination (myelin water fraction) were observed over time in the brain, particularly in the insular cortex.

[0468] Example 2

[0469] Myelination trajectories according to longitudinal studies

[0470] Based on the available data, longitudinal trajectory analysis was performed for two formulas containing different amounts of the identified nutritional driver, sphingomyelin (the composition of these formulas is reported in Table 2 below):

[0471] Table 2 :

[0472]

[0473] The results are Figure 1 Reported in.

[0474] The graph shows that the higher the folate content in the formula, the closer the myelination trajectory of infants fed that formula is to that provided by feeding exclusively on human breast milk.

[0475] Based on the results reported in the experimental section, it has been demonstrated that nutritional compositions comprising vitamins or mixtures thereof determine a trajectory of myelination in the brain that is optimal (close to the trajectory determined by human breastfeeding) in infants fed with such compositions. As discussed above, it is believed that this optimal trajectory in brain myelination corresponds to an effective improvement in the cognitive abilities of infants, matching the abilities of infants fed with breast milk and balancing the deficiencies in those cognitive abilities exhibited in infants fed with formulas low in vitamin A.

[0476] Example 3

[0477] Co-culture of neurons and OLs

[0478] Neurons / oligodendrocytes were cultured as previously described by Charles et al., 2000.

[0479] Pregnant female rats (Wistar rats) at 17 days of gestation were killed by cervical dislocation and the fetuses were removed from the uterus. The forebrain was removed and placed in ice-cold Leibovitz medium (L15) containing 2% penicillin-streptomycin (PS) and 1% bovine serum albumin (BSA). The forebrain was dissociated by trypsinization at 37°C for 20 minutes (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 aspiration and extrusion through a 10 ml pipette 3 times. The cells were then centrifuged at 180×g for 10 minutes at 4°C on a layer of BSA (3.5%) in L15 medium. 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 Neurobasal supplemented with 2% B27, 2mM L-glutamine (L Glu), 2% PS solution, 1% FCS and 10ng / ml platelet-derived growth factor (PDGF-AA). The live cells were counted in a Neubauer cell counter using a trypan blue exclusion test. The cells were seeded at a density of 20,000 cells / well in a 96-well plate pre-coated with poly-L-lysine and laminin.

[0480] One day after seeding (day 1 of culture), cells were incubated with test compounds ( Selected from those listed in Table 3 ) or 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 effect on the early myelination process. The positive effect of estradiol on neurite outgrowth has also been published (for review, please see Alevaro et al., 2010).

[0481] In a humidified incubator, the flat plate is maintained at 37°C in an atmosphere of air (95%)-CO2 (5%). Replace half of the culture medium with fresh culture medium and test compound or control compound every other day. Test compound or control compound maintains a defined concentration during the experiment. Test compound (6 holes for each condition) on 1 culture. Then use cells to measure OPC proliferation, OPC differentiation into OL and early myelination process (myelin sheath wrapping) at the 12th, 18th or 30th day of cultivation, or OL maturation (myelin maturation) and mature myelination process (myelin sheath wrapping).

[0482] Proliferation of OPCs - measurement of A2B5 positive cells and total neurite length (NF)

[0483] On day 12 of culture, cells were fixed with a cold mixture of absolute ethanol (95%) and pure acetic acid (5%) for 5 minutes. Cells were then permeabilized and nonspecific sites were blocked with phosphate buffered saline (PBS) solution containing 0.1% saponin and 1% FCS for 15 minutes at room temperature.

[0484] The cells were then incubated with monoclonal anti-A2B5-conjugated 488 (1 / 200 dilution in PBS containing 1% FCS, 0.1% saponin) was incubated for 2 hours at room temperature and with anti-NF (Neurofilament 200, phosphorylated and non-phosphorylated) produced in rabbit (1 / 500 dilution in PBS containing 1% FCS, 0.1% saponin) for 2 hours at room temperature. The antibody was developed with Alexa Fluor 568 goat anti-rabbit (1 / 400 dilution in PBS containing 1% FCS, 0.1% saponin) for 1 hour at room temperature.

[0485] The total number of OPCs (number of A2B5 positive cells) was quantified (to assess proliferation) and the axonal network (total neurite length (NF)) was measured to assess the effect of the compounds on the neuronal network (the quality of myelination is directly related to the quality of the axonal network).

[0486] Differentiation of OPCs into OLs and myelination process (myelin sheathing) - measurement of the number and area of ​​MAG-positive cells, weight Overlapping MAG / NF wrapping and total neurite length (NF)

[0487] On day 18 of culture, cells were fixed with a cold mixture of absolute ethanol (95%) and pure acetic acid (5%) for 5 minutes. Cells were then permeabilized and nonspecific sites were blocked with phosphate buffered saline (PBS) solution containing 0.1% saponin and 1% FCS for 15 minutes at room temperature.

[0488] The cells were then incubated for 2 hours at room temperature with monoclonal anti-MAG produced in mouse (1 / 400 dilution in PBS containing 1% FCS, 0.1% saponin) and anti-NF (Neurofilament200, phosphorylated and non-phosphorylated) produced in rabbit (1 / 500 dilution in PBS containing 1% FCS, 0.1% saponin). The cells were developed with CF 488A goat anti-mouse (1 / 800 dilution in PBS containing 1% FCS, 0.1% saponin) and Alexa Fluor 568 goat anti-rabbit (1 / 800 dilution in PBS containing 1% FCS, 0.1% saponin) for 1 hour at room temperature.

[0489] The total number of OLs (number and area of ​​MAG positive cells) (to evaluate the differentiation process) and the wrapping of OPCs around axons (overlapping MAG / NF wrapping) (myelination process) were quantified. The axonal network (total axon length (NF)) was measured to evaluate the effect of compounds on the neuronal network.

[0490] OL maturation (myelin maturation) - measure the number and area of ​​MBP-positive cells, overlap

[0491] MBP / NF packaging and total neurite length (NF)

[0492] On day 30 of culture, cells were fixed with a cold mixture of absolute ethanol (95%) and pure acetic acid (5%) for 5 minutes. Cells were then permeabilized and nonspecific sites were blocked with phosphate buffered saline (PBS) solution containing 0.1% saponin and 1% FCS for 15 minutes at room temperature.

[0493] The cells were then incubated for 2 hours at room temperature with monoclonal anti-MBP raised in mouse (1 / 1000 dilution in PBS containing 1% FCS, 0.1% saponin) and anti-NF raised in rabbit (Neurofilament200, phosphorylated and non-phosphorylated) (1 / 500 dilution in PBS containing 1% FCS, 0.1% saponin). The cells were developed for 1 hour at room temperature with CF 488A goat anti-mouse (1 / 800 dilution in PBS containing 1% FCS, 0.1% saponin) and Alexa Fluor 568 goat anti-rabbit (1 / 400 dilution in PBS containing 1% FCS, 0.1% saponin).

[0494] 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)) were assessed. Axonal networks (total neurite length (NF)) were measured to assess the effects of compounds on neuronal networks.

[0495] For all measurements, once the culture was complete (6 wells per condition), 30 images (each representing a field of view) were collected and analyzed per well using ImageXpress (Molecular devices) at 20x magnification with LED lights (excitation wavelengths 360 / 480 / 565 and emission wavelengths 460 / 535 / 620) for each test condition. 30 images were automatically acquired, representing 80% of the total surface of the culture wells.

[0496] Results are presented as the cumulative mean length (in μm) of the neurite network or myelin sheath labeled with a given marker (MAG or MBP) per field of view. The overlapping area between NF and MAG or MBP was measured to assess encapsulation.

[0497] To evaluate the OPC population, MAG-positive cell population, and MBP-positive cell population, the number of positive cells in each image (=field of view) was automatically counted. The results are expressed as the average number of positive cells per field of view.

[0498] All images were acquired under the same conditions.

[0499] Table 3

[0500] Plate 1 (A2B5 / NF) Comparison Estradiol (150nM) DHA(0.15μM) DHA(1.5μM) Stearic acid (50 μM) Stearic acid (5 μM) Stearic acid (0.5 μM) B12(100nM) B12(10nM) B12(1nM) Folic acid (250nM) Folic acid (50nM) Folic acid (6nM) Choline (20 μM) Iron (1μM) Iron (0,1μM) Zinc (5μM) Zinc (0,5 μM) Phosphorus (5 mM) Phosphorus (1mM) Magnesium (25 mM) Copper (0,5mM) Phosphatidylcholine (100 μM) Phosphatidylinositol (5 μM) Phosphatidylinositol (50 μM) Phosphatidylserine (5 μM) Phosphatidylserine (10 μM) Phosphatidylserine (100 μM) Sphingomyelin (5 μM) Sphingomyelin (25 μM) Ceramide (brain extract): DPPC (1:4) Galactosylceramide (C18:1 / 24:1) / (C18:1 / 18:0):DPPC(1:4) Glucosylceramide (C18:1 / 24:1) / (C18:1 / 18:0):DPPC(1:4) D-erythro-dihydroceramide (C24:1 / 18:0) / (C18:0 / 18:1):DPPC(1:4) Ceramide-1-phosphate (C18:1 / 24:0): DPPC (1:4) GM3:DPPC(1:4) GD3:DPPC(1:4)

[0501] The results are shown in Figures 4 to 24 middle.

[0502] Example 4

[0503] Materials and methods

[0504] 1. Feeder Layer Preparation: Dissociation of Neonatal Cortices and Maintenance of Mixed Glial Cell Cultures

[0505] Freshly dissected brains were placed in a 37°C water bath for 3 minutes, and then the cortex was cut into small pieces by a P1000 pipette tip. 75 μL of OPC papain solution was added to each brain, and the tissue was then incubated in a 37°C water bath for 20 minutes. Mixed glial cell culture was then added to the tissue suspension to inactivate the OPC papain solution.

[0506] The tissue was then triturated using a sterile fire-polished glass Pasteur pipette and 4 mL of mixed glial cell culture medium was added per brain. The cells were centrifuged at 1200 rpm (approximately 300 g) for 5 minutes and then resuspended in warm mixed glial cell culture medium and plated in PLL-coated flasks.

[0507] After 4 hours of 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 performed. The cells were then maintained in culture until confluent.

[0508] 2. Preparation of Hippocampal Neurons

[0509] Hippocampal neurons were isolated from embryonic (E18) pups of Sprague Dawley rats. Briefly, after animal sacrifice, the brain was isolated, the meninges were removed from the medial aspect of the cerebral hemispheres, and the hippocampus was dissected out and kept at 4°C until the procedure was completed.

[0510] The tissue was then incubated with 2.5% trypsin in a 37°C water bath for 15 min and then gently washed and kept in culture medium. Hippocampal dissociation was performed by repeatedly pipetting them with a functionalized sterile Pasteur pipette. After mechanical dissociation, cells were plated at the desired density in neuronal plating medium, allowed to recover for 4 h, and then placed in complete neuronal medium.

[0511] 3. Purification of OPCs from Mixed Glial Cell Cultures to Establish OL / Hippocampal Neuron Co-cultures

[0512] On day 9 of the mixed glial cell culture, the flasks were shaken at 50 rpm for 45 minutes on an orbital shaker in a 5% CO2 tissue culture incubator. The purpose of this shaking was to remove any loosely adherent contaminating cells from the monolayer.

[0513] The medium was then changed and replaced with 4 mL of fresh mixed glial medium supplemented with 5 μg / mL insulin. The flask was then placed back on the shaker and equilibrated for approximately 3 hours, followed by shaking at 220 rpm for approximately 16 hours (overnight).

[0514] The next morning, the mixed glial culture medium containing microglia and OPC cells was collected and pre-plated on P100 dishes (untreated) for 30 minutes to purify OPC cells; microglia immediately began to adhere to the dish, while OPC cells remained in the supernatant culture medium.

[0515] 30 min after pre-plating, the culture medium was collected, and OLs were counted and plated on hippocampal neurons in a final volume of 1 mL of OL culture medium.

[0516] A complete OL medium (no CNTF) exchange was performed and cells were then maintained in culture until the appropriate experimental time.

[0517] For the mature experiment, the experimental process is as follows:

[0518] a. OPCs were cultured in vitro on astrocyte feeder layers for 10 days

[0519] b. Isolation of OPCs (Day 0)

[0520] c. Administration of compound (Day 3)

[0521] d. Quantitative assessment of maturity on days 4, 7 and 10.

[0522] For the myelination experiment, the experimental procedure was as follows:

[0523] a. Cultivate hippocampal neurons until the neural network is fully mature (14 days in vitro)

[0524] b. At the same time, OPCs were cultured in vitro on astrocyte feeder layers for 10 days

[0525] c. Isolation of OPCs and co-culture with neurons (Day 14)

[0526] d. Administration of compound (Day 15)

[0527] e. On day 15 of culture (one day after co-culture plating and before compound treatment),

[0528] Quantitative evaluation of myelination on days 18, 21 / 23, and 28 / 29

[0529] 4. Image Acquisition

[0530] All cultures at different experimental time points were fixed in 4% paraformaldehyde and 4% sucrose for 10 minutes at room temperature (RT). Primary and secondary antibodies were applied to 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. 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 acquired using Array Scan XTI (ThermoScientific); objective was 20x, using 2x2 binning mode. For each condition and replicate well (triplicate), at least 15 images were acquired.

[0531] To analyze all acquired images, HCS Studio Cell Analysis Software was used, specifically the "Scan" application.

[0532] OPC papain solution (made in MEM)

[0533] Papain solution 1.54 mg / mL

[0534] L-cysteine ​​360 ​​μg / mL

[0535] DNase I 60 μg / mL

[0536] Mixed glial cell culture medium (made in DMEM)

[0537] FBS10%

[0538] Pen / Strep (0.33%, from stock solution) 33 units / mL of penicillin and 33 μg / mL of streptomycin

[0539] GlutaMAX 1%

[0540] OL medium

[0541] DMEM

[0542] 100X OL-Supplement

[0543] Bovine insulin (from 1 mg / mL stock solution)

[0544] GlutaMAX

[0545] Homoferric transferrin (from 33 mg / mL stock solution)

[0546] B27 Supplements

[0547] FBS

[0548] CNTF (from 50 ng / μL stock solution)

[0549] The results are shown in Figure 26 to Figure 45 middle.

Claims

1. A nutritional composition for infants and / or young children, comprising vitamins or mixtures thereof for promoting and / or supporting an optimal myelination trajectory in the brain.

2. The composition of claim 1, wherein the optimal myelination trajectory in the brain approximates the myelination trajectory observed in infants fed exclusively with human breast milk in the early months after birth.

3. A 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 prevention of neurocognitive deficits.

4. A composition according to any one of claims 1 to 3, comprising folic acid in an amount higher than 50 mcg / 100 g and / or vitamin B12 in an amount higher than 0.01 mcg / 100 g.

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

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

7. The composition according to any one of claims 1 to 6, further comprising sphingomyelin.

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

9. Use of a composition as claimed in any one of claims 1 to 8 for promoting and / or supporting an optimal myelination trajectory in the brain of an infant or young child.

10. Use of a composition as claimed in any one of claims 1 to 8 for preventing a suboptimal myelination trajectory in the brain of an infant or young child.

11. A method for promoting and / or supporting an optimal myelination trajectory in the brain of an infant and / or young child, the method comprising administering to the infant and / or young child a nutritional composition comprising folic acid.

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

13. A nutritional composition, comprising: Sphingomyelin in an amount greater than 200 mg / kg; Iron in amounts above 5 mg / 100 g; Choline in an amount greater than 30 mg / 100 g; DHA in an amount higher than 30 mg / 100 g; Folic acid in an amount greater than 50mcg / 100g; and or vitamin B12 in an amount greater than 0.01mcg / 100g.

14. A nutritional composition, comprising: Sphingomyelin in an amount ranging from 200 mg / kg to 2 g / kg; Iron in an amount ranging from 5 mg / 100 g to 40 mg / 100 g; Choline in an amount ranging from 30 mg / 100 g to 1000 mg / 100 g; DHA in an amount ranging from 30 mg / 100 g to 300 mg / 100 g; Folic acid in an amount ranging from 50mcg / 100g to 500mcg / 100g; Vitamin B12 in an amount greater than 0.1mcg / 100g to 10mcg / 100g.

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