Nutritional composition for promoting development of central nervous system and application

Abstract

The invention discloses a nutritional composition for promoting central nervous system development and application. The invention provides a nutritional composition for promoting the development of a central nervous system. The nutritional composition comprises the following necessary components: a breast milk oligosaccharide component and a casein phosphopeptide component, moreover, in the composition, the mass ratio of the breast milk oligosaccharide component to the casein phosphopeptide component is (1-100): (500-1). According to the invention, the breast milk oligosaccharide component and the casein phosphopeptides are matched to prepare the nutritional composition, and the nutritional composition can significantly promote nerve development such as neuronal maturation, synaptic generation and myelin sheath formation.

Description

Technical Field

[0001] This invention relates to a nutritional composition, belonging to the food field, and more specifically, to a nutritional composition and its application that promotes the development of the central nervous system. Background Technology

[0002] Early life nutrition has been shown to play a crucial role in neurodevelopment, such as neuronal maturation, synapsis, and myelination. Myelination is the process by which specialized glial cells in the central nervous system (CNS)—oligodendrocytes (OLs)—form a myelin sheath around an axon, which is essential for normal brain connectivity. The myelin sheath consists of several condensed lipid bilayer membranes. It increases axonal conduction velocity by reducing the capacitance of the axonal membrane and allows jumping currents. Brain myelination parallels the maturation of cognitive function. In humans, myelination begins in mid-pregnancy, peaks in the first few years of life, and continues into adulthood.

[0003] Oligodendrocyte progenitor cells (OPCs) are the main glial cell population in the central nervous system, accounting for 2%-9% of the total cell population. OPCs specifically express tetrasialotetrahexosylganglioside (GOlc) on their surface, which can be recognized by the monoclonal antibody A2B5. After mitosis, OPCs differentiate into myelinated oligodendrocytes (OLs). These OLs undergo numerous processes, establishing contact with the axons of different neurons and initiating myelination. During their maturation, OLs produce different components of myelin, such as lipids (cholesterol, galactolipids, and phospholipids) and myelin-specific proteins. The types of myelin proteins expressed by OLs, such as myelin-associated glycoprotein (MAG) and myelin-binding protein (MBP), are associated with their maturation period. Myelinated OLs express MAG, and MAG expression gradually increases during OL maturation. MAG is a sialic acid-binding immunoglobulin-like lectin. Although it constitutes only a small fraction of the total myelin protein content, it is primarily expressed in the peri-axonal region of the myelin sheath. It appears to play a crucial role in oligodendrocyte-axon interactions and mediates bidirectional signaling between axons and oligodendrocytes (OLs) to support myelin formation. MBP is expressed in mature myelinated OLs and is one of the major components of myelin. MBP appears to play an active role in myelin formation and compaction. In fact, MBP polymerizes and forms a viscous reticular protein network, which is essential for hopping currents.

[0004] In the central nervous system, every step of myelination, including the proliferation of OPCs, the differentiation and maturation of OPCs into myelinated OLs, and myelination itself, is highly regulated by both external and internal factors. In particular, different nutrients have varying effects on myelination, suggesting that early life nutrition may play a significant role in its regulation. Therefore, identifying early life nutrition factors that support myelination is crucial for optimal brain and cognitive development.

[0005] Existing technologies have conducted some research on nutritional compositions that promote brain development, for example:

[0006] Reference 1 relates to a nutritional composition comprising medium- and long-chain fatty acid triglycerides and human milk oligosaccharides, wherein the mass ratio of medium- and long-chain fatty acid triglycerides to human milk oligosaccharides is 1:1 to 9:1, which has the effect of improving brain development and promoting neural and / or cognitive development.

[0007] Reference 2 relates to the application of a nutritional composition in the preparation of products that promote brain development. The nutritional composition includes whey protein powder, casein glycomacropeptide, and sialic acid, wherein the mass ratio of whey protein powder, casein glycomacropeptide, and sialic acid is (1-30):(1-3.5):1.

[0008] The above-mentioned references 1-2 only evaluated the effects of their compositions on spatial learning and memory abilities, and did not involve content related to promoting brain neural development, such as the growth and differentiation of brain nerve cells, the construction of neural networks, and the regulation of neurotransmitters.

[0009] It can be seen that although some research has been conducted on nutritional compositions that promote neural development in the existing technology, it cannot be said that they are sufficient or perfect. There is still a need to develop effective nutritional compositions that promote the development of the central nervous system and / or help improve memory.

[0010] Human milk oligosaccharides (HMOs) are a class of complex carbohydrates naturally found in breast milk, composed of 3-10 monosaccharide molecules linked together in specific ways. There are over 150 structural variations of HMOs, all originating from a basic lactose unit, which is extended and modified, such as through focusing or sialylation, to produce different subgroups. Current research on HMOs mainly focuses on preparation methods, their efficacy in preventing and treating diseases, applications in nutritional compositions, and detection methods. Among these, applications in compositions primarily focus on immune and inflammatory aspects.

[0011] Casein is a phosphorus- and calcium-binding protein. Current research on casein mainly focuses on its immune function and its role in promoting calcium and other ion absorption. Casein hydrolysate peptides are bioactive peptides produced after casein undergoes a hydrolysis reaction. They have the effects of supplementing protein, promoting tissue and cell growth and development, promoting bone health, regulating and enhancing immune function, and improving the body's ability to fight viruses and bacteria.

[0012] However, in existing research, the mechanism of action, actual effects, and degree of efficacy of the combined use of human milk oligosaccharides and casein phosphopeptides in promoting neurodevelopment remain unclear. Studies on the brain development of this combination are limited to passive memory studies using animal experiments, without exploring the development of neurons or investigating the effects of specific dosages on early brain development. It has not yet been definitively revealed how these two components synergistically influence the development of the central nervous system when present in a specific combination, nor has their effect on cognitive enhancement been clarified.

[0013] References

[0014] Reference 1 CN119949517A

[0015] Reference 2CN118252257A Summary of the Invention

[0016] The problem the invention aims to solve

[0017] As mentioned earlier, there is still room for further research and development regarding nutritional compositions that promote brain development. Furthermore, current research on brain development compositions is limited to passive memory studies using animal experiments. There is an urgent need to provide a nutritional composition that can effectively promote myelin development, neuronal maturation, synapse formation, and other processes that promote the development of the central nervous system and enhance cognitive function. This nutritional composition is crucial for the normal growth, functional maintenance, and damage repair of the nervous system. In the field of neurodevelopmental nutritional compositions, myelin formation is essential for the normal functioning of the nervous system. References 1-2 do not address content related to promoting myelin development, neuronal maturation, and synapse formation, or other processes that promote the development of the central nervous system.

[0018] Therefore, there is a need for a food that provides a nutritional composition that can promote the development of the central nervous system, such as myelin formation, and is also absorbable and usable in order to better exert its promoting effect on the development of the central nervous system and brain function.

[0019] In view of this, the present invention provides a nutritional composition comprising a human milk oligosaccharide component and a casein phosphopeptide component, which significantly promotes central nervous system development, including myelin sheath development and formation. Specifically, the present invention explores the effects of the composition on promoting central nervous system development such as myelin cell proliferation, differentiation, and synapse formation, and studies the influence of the composition and its component ratios on central nervous system development. By scientifically designing the ratio of human milk oligosaccharides and casein phosphopeptides in the composition, a synergistic effect can be achieved, meeting market demand for functional compositions. This composition for promoting central nervous system development provided by the present invention is particularly suitable for people of different age groups, providing a safe and effective nutritional composition for promoting healthy central nervous system development and further enhancing various brain functions, including cognitive function.

[0020] Solution for solving the problem

[0021] [1]. A nutritional composition for promoting the development of the central nervous system, wherein the nutritional composition comprises the following essential components: a human milk oligosaccharide component and a casein phosphopeptide component; and, in the nutritional composition, the mass ratio of the human milk oligosaccharide component to the casein phosphopeptide component is (1-100):(500-1).

[0022] [2]. The nutritional composition according to [1], wherein the human milk oligosaccharide component comprises one or more of 3'-sialylated lactose, 6'-sialylated lactose, 4'-galactosyllactose, 3'-galactosyllactose, 2'-fucosylated lactose, lactose-N-tetrasaccharide, lactose-difucosylated tetrasaccharide, lactose-N-neotetrasaccharide, 6'-galactosyllactose and lactose-N-fucopentose-I, preferably, the human milk oligosaccharide component comprises 3'-sialylated lactose (3'-SL) and 6'-sialylated lactose (6'-SL).

[0023] [3]. According to the nutritional composition described in [2], the mass ratio of 3'-sialylated lactose (3'-SL) and 6'-sialylated lactose (6'-SL) in the human milk oligosaccharide component is (1-15):(15-1).

[0024] [4]. The composition according to any one of [1] to [3], wherein the mass ratio of 3'-sialylated lactose (3'-SL) and 6'-sialylated lactose (6'-SL) in the human milk oligosaccharide component is (1 to 10):(10 to 1), preferably (1 to 5):(5 to 1).

[0025] [5]. A food product, wherein the food product contains or uses any of the nutritional compositions described in [1] to [4].

[0026] [6]. The food according to [5], wherein the food is any one or more of infant food, children's food, adolescent food, pregnant and postpartum food, adult food and middle-aged and elderly food.

[0027] [7]. The food according to [5] or [6], wherein the food may be in solid, semi-solid or liquid form.

[0028] [8]. Application of compositions containing two essential components, human milk oligosaccharides and casein phosphopeptides, in the preparation of nutritional compositions that promote the development of the central nervous system and / or help improve memory.

[0029] [9]. Application of compositions containing two essential components, human milk oligosaccharides and casein phosphopeptides, in the preparation of foods that promote the development of the central nervous system and / or help improve memory.

[0030]

[10] . According to the use described in [8] or [9], wherein the development of the central nervous system includes brain development; preferably, the development of the central nervous system includes at least one of neuronal maturation, synapsis and myelination.

[0031]

[11] . According to the use described in

[10] , the development of the central nervous system includes promoting the proliferation of oligodendrocytes, promoting the myelination of oligodendrocytes, and promoting the maturation of oligodendrocytes.

[0032]

[12] . According to the use described in

[10] , wherein the development of the central nervous system includes promoting the expression of synapse-related proteins, preferably, the synapse-related proteins include at least one of synaptophysin (SYN), postsynaptic density protein 95 (PSD 95), brain-derived neurotrophic factor (BDNF) and growth-associated protein-43 (GAP-43).

[0033] The effects of the invention

[0034] 1) The present invention provides a nutritional composition comprising a human milk oligosaccharide component and a casein phosphopeptide component, which can promote the development of the central nervous system.

[0035] 2) The composition provided by this invention can promote the development of the central nervous system. Specifically, the composition provided by this invention has broad application prospects in improving memory and promoting the development of the central nervous system. In the process of central nervous system development, it plays an important role in controlling various cellular mechanisms such as neuronal maturation, synapse formation, and myelination.

[0036] 3) In in vitro models, cells treated with the composition provided by the present invention increased the number of OPCs, promoted differentiation or maturation into OLs and the myelination properties of OLs, and the components had a good synergistic effect.

[0037] 4) The human milk oligosaccharide component and casein peptide component, especially the 3'-SL, 6'-SL and casein phosphopeptide components, in the composition of the present invention, when used in limited amounts, can achieve a synergistic effect, promote neuronal maturation and synapse formation, further increase the expression of proteins related to oligodendrocyte maturation and differentiation, and promote the expression of myelin maturation-related proteins in brain tissue.

[0038] 5) When used in a limited amount, the composition of the present invention has a synergistic effect, promotes neuronal maturation and synapse formation, and can also significantly promote the increase of axon density and synapse-related protein expression in the hippocampus and cortex, further enhancing synaptic stability and transmission efficiency, and improving the efficient information transmission of the nervous system.

[0039] 6) The human milk oligosaccharide component and casein peptide component in the composition of the present invention, when used in a limited amount, can help improve memory. Specifically, they can increase the content of metabolites that promote the formation of neurite buds and synapses in brain tissue, thereby further enhancing the memory-improving effect.

[0040] 7) The composition of the present invention has a wide range of applications and is suitable for various groups of people such as infants, children, adolescents, pregnant women, adults and the elderly. It is also convenient to add to or prepare various foods or health products. Attached Figure Description

[0041] Figure 1A The effect of different nutrient composition samples on nerve cells cultured in vitro for 12 days is shown in the A2B5 immunostaining image (green). Scale bar: 50 μm; numbers in the figure indicate experimental groups.

[0042] Figure 1B The effect of different nutrient composition samples on nerve cells cultured in vitro for 12 days is shown in the A2B5 immunostaining image (green). Scale bar: 50 μm; numbers in the figure indicate experimental groups.

[0043] Figure 2 The effect of different nutrient composition samples on nerve cells cultured in vitro for 18 days is shown in the MAG immunostaining image (green). Scale bar: 50 μm; numbers in the figure indicate experimental groups.

[0044] Figure 3 The effect of different nutrient composition samples on nerve cells cultured in vitro for 30 days is shown in the MBP immunostaining image (green). Scale bar: 50 μm; numbers in the figure indicate experimental groups.

[0045] Figure 4 The image shows a magnetic resonance T1-weighted image; the numbers in the image indicate the experimental group.

[0046] Figure 5 The magnetic resonance DTI image is shown; the numbers in the image indicate the experimental group.

[0047] Figure 6 The magnetic resonance (MRS) image is shown; the numbers in the image indicate the experimental group. Detailed Implementation

[0048] Various exemplary embodiments, features, and aspects of the present invention will be described in detail below. The term "exemplary" as used herein means "serving as an example, embodiment, or illustration." Any embodiment described herein as "exemplary" is not necessarily to be construed as superior to or better than other embodiments.

[0049] Furthermore, to better illustrate the present invention, numerous specific details are set forth in the following detailed embodiments. Those skilled in the art should understand that the present invention can be practiced without certain specific details. In other instances, methods, means, apparatus, and steps well known to those skilled in the art have not been described in detail in order to highlight the spirit of the present invention.

[0050] Unless otherwise stated, all units used in this specification are international standard units, and all numerical values ​​and ranges appearing in this invention should be understood to include systematic errors that are unavoidable in industrial production.

[0051] In this specification, the word "may" has two meanings: to perform a certain process and not to perform a certain process.

[0052] In this specification, references to "some specific / preferred embodiments," "other specific / preferred embodiments," "implementation," etc., refer to specific elements (e.g., features, structures, properties, and / or characteristics) related to that embodiment, which are included in at least one of the embodiments described herein and may or may not be present in other embodiments. Furthermore, it should be understood that these elements may be combined in any suitable manner in various embodiments.

[0053] In this specification, the range of values ​​referred to as "value A to value B" refers to the range including the endpoint values ​​A and B.

[0054] In this invention, the term "about" is used to define that the numerical ranges and parameters of this invention are approximate values, while specific related values ​​have been presented as precisely as possible. Unless otherwise explicitly stated, it should be understood that all ranges, quantities, values, and percentages used in this invention are modified by "about". Here, "about" generally means that the actual value is within ±3%, ±2%, ±1%, or ±0.5% of a specific value or range.

[0055] In this invention, the term "infant" is used to refer to the human group aged 0 to 6 months.

[0056] In this invention, the term "older infant" refers to the human group aged 6 to 12 months.

[0057] In this invention, the term "infant" is used to refer to the human group aged 12 to 36 months.

[0058] In this invention, the term "infant" refers to the human group under the age of 3 years.

[0059] In this invention, the term "children" refers to a group of human beings who are older than 3 years and younger than 12 years and are in the growth and development stage.

[0060] In this manual, the term "adult" refers to a person who is 18 years of age or older.

[0061] In this manual, the term "teenager" refers to people aged 7-40.

[0062] In this manual, the term "middle-aged person" refers to a person aged 41-65.

[0063] In this manual, the term "elderly person" or "senior citizen" refers to a person aged 65 or older. In this manual, the numerical range indicated by "above" or "below" refers to a range that includes the stated number.

[0064] In this specification, the terms "optional" or "optional" are used to indicate the use or omission of certain substances, components, procedures, application conditions, etc.

[0065] <First Aspect>

[0066] In a first aspect, the present invention provides a nutritional composition comprising the essential components of a human milk oligosaccharide component and a casein phosphopeptide component, wherein the mass ratio of the human milk oligosaccharide component to the casein phosphopeptide component in the composition is (1-100):(500-1). This nutritional composition exhibits a significant synergistic effect and can promote the development of the central nervous system, such as promoting myelin formation.

[0067] Compared to individual components of human milk oligosaccharides and casein phosphopeptides, the nutritional composition provided by this invention can synergistically promote central nervous system development and / or aid in memory improvement, such as promoting myelin formation. The effects of promoting central nervous system development and / or aiding in memory improvement are particularly pronounced when the ratio of the human milk oligosaccharide and casein phosphopeptide components is within the aforementioned range.

[0068] The technical solution of this invention is mainly based on the following insights:

[0069] This invention first verified, from a (cell)biological perspective, the range within which human milk oligosaccharide and casein phosphopeptide components can promote myelin nerve development. Within this range, the biological performance of each component individually, as well as in different combinations, was studied. Unexpectedly, the combination of human milk oligosaccharide and casein phosphopeptide was discovered. Then, from an (animal)biological perspective, it was verified that the ratio of human milk oligosaccharide and casein phosphopeptide components is a safe dosage, especially at specific ratios, which have the effect of promoting central nervous system development and / or assisting in improving memory.

[0070] (Human milk oligosaccharide components)

[0071] Human milk oligosaccharides (HMOs) are a collective term for oligosaccharides with a degree of polymerization ≥3 that are naturally found in human milk. They are formed by modifying the terminal positions of lactose molecules with five monomers: glucose (Glc), galactose (Gal), N-acetylglucosamine (GlcNAc), fucose (Fuc), and N-acetylneuraminic acid (Neu5Ac). Each HMO molecule contains 3 to 32 monosaccharides linked by different glycosidic bonds, contributing to the diversity and complexity of HMOs.

[0072] HMOs are mainly composed of five core monomers: glucose, sialic acid, fucose, N-acetylglucosamine, and galactose. Different HMOs exhibit different fucosylation and sialylation, thus HMOs in breast milk can be classified into neutral fucosylated HMOs, acidic sialylated HMOs, and neutral non-fucosylated HMOs.

[0073] Currently, the main methods for producing human milk oligosaccharides include chemical synthesis, enzymatic synthesis, and bioengineering synthesis. At the same time, these components can also be indirectly introduced from various existing milk components.

[0074] It should be noted that the use of the various human milk oligosaccharides described above in this invention shall comply with the requirements of local laws and regulations. In some cases, where permitted by laws and regulations, these components may be directly introduced into the food composition as individual raw materials; in other cases, where permitted by laws and regulations, they may be indirectly introduced into the composition or food through the addition of qualified dairy raw materials.

[0075] This invention does not specifically limit the source of human milk oligosaccharides; typically, they can be derived from various milk-containing raw materials, such as animal milk raw materials (e.g., cow's milk, milk powder, etc.). In this invention, "animal milk" is used to refer to the liquid obtained from the mammary glands of mammals in lactation.

[0076] The term “animal milk” should be interpreted broadly and encompass both raw milk (i.e., the liquid obtained directly from the mammary glands) and standardized dairy products.

[0077] In some embodiments, the human milk oligosaccharides of the present invention are one or more of 3'-sialyl lactose, 6'-sialyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose, 2'-fucosyl lactose, lactose-N-tetrasaccharide, lactose-difucosyltetrasaccharide, lactose-N-neotetrasaccharide, 6'-galactosyl lactose, lactose-N-fucopentose-I, and others.

[0078] In some preferred embodiments, the human milk oligosaccharide component comprises 3'-sialylated lactose (3'-SL) and 6'-sialylated lactose (6'-SL).

[0079] In some embodiments, the mass ratio of 3'-sialylated lactose (3'-SL) to 6'-sialylated lactose (6'-SL) in the human milk oligosaccharide component is (1-15):(15-1).

[0080] In some preferred embodiments, the mass ratio of 3'-sialylated lactose (3'-SL) to 6'-sialylated lactose (6'-SL) in the human milk oligosaccharide component is (1-10):(10-1). For example, the mass ratio of 3'-sialylated lactose (3'-SL) to 6'-sialylated lactose (6'-SL) is 5:2, 1:2, 1:10, etc.

[0081] In some more preferred embodiments, the mass ratio of 3'-sialylated lactose (3'-SL) to 6'-sialylated lactose (6'-SL) in the human milk oligosaccharide component is (1-5):(5-1).

[0082] In some further synergistic embodiments, the mass ratio of 3'-sialylated lactose (3'-SL) to 6'-sialylated lactose (6'-SL) in the human milk oligosaccharide component is 1:2 and / or 5:2. In some exemplary embodiments, the mass ratio of 3'-sialylated lactose (3'-SL) to 6'-sialylated lactose (6'-SL) may be 1:2.

[0083] The human milk oligosaccharide component of the present invention can exist in a liquid state, or in a semi-solid or solid state.

[0084] When the human milk oligosaccharide component of the present invention is present in a liquid form, in some preferred embodiments, the concentration of the human milk oligosaccharide component in the nutritional composition is 0-5 mg / mL.

[0085] In some preferred embodiments, the concentration of the human milk oligosaccharide component is 0-3 mg / mL. In some further preferred embodiments, the concentration of 3'-sialylated lactose (3'-SL) in the nutritional composition is less than 1 mg / mL, and the concentration of 6'-sialylated lactose (6'-SL) is less than 1.5 mg / mL. More preferably, the concentration of 3'-sialylated lactose (3'-SL) is 0.01 mg / mL-0.5 mg / mL, and the concentration of 6'-sialylated lactose (6'-SL) is 0.01 mg / mL-1 mg / mL, for example: the concentration of 3'-SL is 0.01 mg / mL; 0.05 mg / mL; 0.25 mg / mL; 0.5 mg / mL, and the concentration of 6'-SL is 0.02 mg / mL; 0.1 mg / mL; 1 mg / mL.

[0086] When the 3'-sialylated lactose and 6'-sialylated lactose in the human milk oligosaccharide component are used in the above ratio and amount, they can play a synergistic role. When used in combination with the casein phosphopeptide component, the nutritional composition can achieve better synergistic effect in promoting the development of the central nervous system.

[0087] (Casein phosphopeptide component)

[0088] In this invention, the casein phosphopeptide component includes casein phosphopeptide (CPP).

[0089] In this specification, casein is a phosphorus- and calcium-bound protein that is sensitive to acid and precipitates at low pH levels. Casein is the main protein in the milk of mammals, including cows, sheep, and humans, and is also known as casein, casein gluten, or lactose.

[0090] In this specification, casein phosphopeptide (also referred to as casein peptide in this invention) refers to the hydrolysate obtained from casein and / or milk protein (optionally separated and purified) by biotechnological means such as enzymatic hydrolysis.

[0091] There are no particular limitations on the source of the casein phosphopeptide of this invention. For example, it can be a bioactive polypeptide derived from various animal milk raw materials through biotechnology. Animal milks such as cow's milk, sheep's milk, horse's milk, and camel's milk are based on cow's milk casein. The CPP molecule consists of twenty to thirty amino acid residues, including four to seven clustered phosphoseryl groups.

[0092] Similarly, the casein phosphopeptide of the present invention can be prepared in-house or obtained commercially.

[0093] (Synergistic effect)

[0094] In this invention, the combined use of human milk oligosaccharides and casein phosphopeptides unexpectedly revealed a synergistic effect in promoting central nervous system development and / or aiding in memory improvement.

[0095] To obtain a more synergistic composition for promoting central nervous system development and / or aiding in memory improvement, in some embodiments, the mass ratio of the human milk oligosaccharide component to the casein phosphopeptide component in the nutritional composition is (1-100):(500-1).

[0096] In some preferred embodiments, the mass ratio of the human milk oligosaccharide component to the casein phosphopeptide component in the nutritional composition is (1-50):(400-1).

[0097] To achieve further and better synergistic effects in promoting central nervous system development and / or aiding in memory improvement, in some more preferred embodiments, the mass ratio of the human milk oligosaccharide component to the casein phosphopeptide component in the nutritional composition is (1-20):(350-1).

[0098] In some exemplary embodiments, the mass ratio of the human milk oligosaccharide component to the casein phosphopeptide component in the nutritional composition is: 15:35; 15:3.5; 1.5:35; 3:350.

[0099] When the human milk oligosaccharide component and casein phosphopeptide are within the above-mentioned range, the composition further achieves a better synergistic effect in promoting the development of the central nervous system and / or assisting in improving memory.

[0100] (Composition)

[0101] The nutritional composition of the present invention includes at least the human milk oligosaccharides and casein phosphopeptides described above. There are no particular limitations on the formation method of the composition; it can be formed by dissolving and mixing the substances containing these components as described above in a solvent, or by mixing two high-purity extracts.

[0102] Furthermore, there are no particular limitations on other components that can be used in the compositions of the present invention. Other edible ingredients, food additives, or solvent components commonly used in the art can be used without impairing the effects of the present invention.

[0103] The nutritional composition of the present invention may exist in a liquid state, or in a semi-solid or solid state.

[0104] When the nutritional composition of the present invention is present in a liquid form, in some embodiments, the concentration of the human milk oligosaccharide component in the nutritional composition is 0-5 mg / mL, preferably, the concentration of the human milk oligosaccharide component in the nutritional composition is 0-2 mg / mL, for example: the concentration of the human milk oligosaccharide component is 0.03 mg / mL; 0.15 mg / mL; 1.5 mg / mL.

[0105] In some embodiments, the concentration of the casein phosphopeptide component in the nutritional composition is less than 50 mg / mL, preferably not exceeding 40 mg / mL. For example, the concentration of the casein phosphopeptide component is 0.035 mg / mL; 0.35 mg / mL; 3.5 mg / mL; 35 mg / mL.

[0106] In some preferred embodiments, when the concentrations of the human milk oligosaccharide component and the casein phosphopeptide component in the nutritional composition meet the above-mentioned ranges, they can synergistically enhance each other and have the effect of promoting the development of the central nervous system and / or assisting in improving memory.

[0107] To further achieve better synergistic effects, in some more preferred embodiments, the concentration of the human milk oligosaccharide component in the nutritional composition is 0.1-1.5 mg / mL, and the concentration of the casein phosphopeptide component is 0.35-35 mg / mL, for example, the concentration of the human milk oligosaccharide component is 0.15 mg / mL; 1.5 mg / mL; and the concentration of the casein phosphopeptide component is 0.35 mg / mL; 35 mg / mL.

[0108] When the concentrations of human milk oligosaccharide components and casein phosphopeptide components meet the above ranges, they exhibit superior synergistic effects and can promote the development of the central nervous system and / or help improve memory.

[0109] (use)

[0110] Furthermore, the above-mentioned composition of the present invention has been experimentally verified to promote the proliferation of ganglion precursor cells, promote myelination of oligodendrocyte precursor cells, promote the expression of myelin-related proteins and / or promote the expression of myelin-related proteins, promote the expression of synaptic-related proteins, increase the content of myelin in the hippocampus and cortex, and promote the production of metabolites glutamine and acetylaspartate. Therefore, the composition can have the effect of promoting brain nerve development and / or assisting in improving memory, and can be used as a food, functional health product or functional health additive.

[0111] <Second aspect>

[0112] In a second aspect of the present invention, a food product is provided, the food product comprising or using the nutritional composition described in the first aspect above.

[0113] The food products of this invention can be suitable for infants, adolescents, adults and / or middle-aged and elderly people to promote the development of the central nervous system and / or help improve memory.

[0114] The food products described in this invention include any one or more of the following: infant food, children's food, adolescent food, pregnant and postpartum food, adult food, and food for the middle-aged and elderly.

[0115] In some specific embodiments, the food can be obtained by processing the nutritional composition according to the first aspect described above.

[0116] There are no particular limitations on the food products of this invention; they can generally be pasta, beverages, instant foods, baked goods, sauces, or functional nutritional supplements.

[0117] For pasta-based foods, this can include staple foods made from flour-based raw materials, as well as whole grain staple foods, such as steamed buns, pancakes, noodles, and filled staple foods.

[0118] For baked goods, this can be cakes or cookies that are mainly based on butter, eggs, and baking powder.

[0119] There are no particular restrictions on beverages; they can include fruit drinks, vegetable drinks, milk tea drinks, tea drinks, milk, yogurt, vitamin drinks, etc. For fruit or vegetable drinks, in addition to including the functional components of this invention, they may also include fruit or vegetable juices or their solid components. For vitamin drinks, in addition to including the two functional components of this invention, they mainly contain various functional vitamins and other functional ingredients, specifically including, for example, white sugar, cyclamate, acesulfame potassium, taurine, potassium sorbate, lysine, inositol, vitamin PP, vitamin B6, vitamin B12, citric acid, etc.

[0120] For reconstituteable foods, typical examples are reconstituteable milk powder products, such as infant formula, adult milk powder, and milk powder for middle-aged and elderly people.

[0121] In addition, there are no particular restrictions on functional nutritional supplements, which can be used as nutritional supplements or meal replacements. Such foods may include one or more of the following components in addition to the two components of this invention: plant product ingredients, animal dairy product ingredients, animal meat product ingredients, protein ingredients, vitamin supplements, mineral supplements, nucleotide supplements, polyunsaturated fatty acid supplements, and any food-acceptable excipients.

[0122] Plants or plant extracts, including fruits such as fig, pomegranate, kiwi, orange, tangerine, pineapple, strawberry, apple, rubber, grape, pear, cherry, blueberry, blackberry, blackcurrant, cranberry, raspberry, melon, amla, and bilberry, or their extracts; fruits and vegetables such as onion, cucumber, tomato, cauliflower, carrot, spinach, kale, Brussels sprouts, garlic, basil, and oregano, or their extracts; grains such as rice (indica, japonica, glutinous rice), cereals (wheat, barley, oats, rye), corn, sorghum, millet, foxtail millet, yellow millet, buckwheat, soybean, broad bean, pea, mung bean, red bean, and kidney bean, or their extracts; nuts such as walnut, pistachio, cashew, hazelnut, almond, apricot kernel, pine nut, peanut, sunflower seed, chestnut, macadamia nut, and ginkgo, or their extracts; and coffee or its extracts.

[0123] Animal-derived ingredients, including meat products from cattle, sheep, fish, or poultry.

[0124] The fat component may include at least one of saturated fatty acids, polyunsaturated fatty acids, monounsaturated fatty acids, OPO structured lipids, DHA, EPA, ARA, and phospholipids. More specifically, the fat includes safflower seed oil, walnut oil, peanut oil, corn oil, soybean oil, argan oil, olive oil, tea oil, sacha inchi oil, coconut oil, perilla oil, deep-sea fish oil, cocoa butter, palm oil, tallow, cream, lard, medium-chain triglycerides, or lecithin, etc.

[0125] Functional additives include vitamins (one or more of vitamin A, beta-carotene, vitamin D3, vitamin E, vitamin K1, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, pantothenic acid, folic acid, niacin, choline, inositol, and biotin), starch, modified starch, amino acids (L-lysine-L-glutamic acid, L-glutamic acid, L-arginine, L-tryptophan, L-glutamine, taurine, L-valine, L-isoleucine, or L-leucine, etc.), traditional Chinese medicine or its extracts, and dietary fiber (inulin, konjac flour, galactooligosaccharides, fructooligosaccharides, isomaltooligosaccharides, soybean polysaccharides, cyclodextrin, resistant dextrin, or soybean fiber, etc.).

[0126] The protein component is selected from at least one of whey protein powder, soy protein isolate, whole milk powder, whole egg powder, lactoferrin, bovine colostrum, amino acids, and protein peptides; and the amino acid is selected from at least one of L-lysine, L-glutamic acid, L-arginine, L-tryptophan, L-glutamine, taurine, L-valine, L-isoleucine, and L-leucine; the protein peptide is selected from one or more of soy oligopeptides, wheat protein peptides, silkworm pupa protein peptides, marine fish oligopeptide powder, cola peptides, amino peptides, and ovalbumin peptides.

[0127] Micronutrient supplements may include metal ion salts of organic acids, such as one or more of the following: calcium citrate, L-calcium lactate, calcium hydrogen phosphate, potassium gluconate, sodium citrate, ferrous gluconate, potassium iodide, zinc gluconate, sodium selenite, copper gluconate, chromium sulfate, manganese gluconate, and magnesium gluconate.

[0128] Any food additives that are acceptable, including but not limited to solvents, antioxidants, antibacterial agents, thickeners, diluents, solubilizers, stabilizers, emulsifiers, fillers, disintegrants, lubricants, coating materials, anti-caking agents, flavoring agents, sweeteners, food flavorings, and food colorings.

[0129] There is no particular limitation on the specific form of the nutritional / health food of the present invention, which can be solid (powder or block, etc.), semi-solid (soft, paste or thick substance, etc.) or liquid.

[0130] This invention does not specifically limit the types of foods that can be prepared using the above-mentioned nutritional compositions and have non-therapeutic effects such as promoting the development of the central nervous system and / or assisting in improving memory, for example, promoting the development of the central nervous system and myelin sheath nerves and assisting in improving memory.

[0131] In some implementations, the food is a solid dairy product or a liquid dairy product; for example, milk powder, cheese, yogurt, liquid milk, etc.

[0132] In some specific implementations, the nutritional or health food can be a powdered reconstituted food (solid beverage, milk powder, instant coffee, cereal powder, nut powder or lotus root powder, etc.) or a liquid beverage (carbonated beverage, fruit and vegetable juice beverage, functional beverage, tea beverage, milk beverage or alcoholic beverage, etc.).

[0133] Typically, the aforementioned foods include: infant formula, maternal formula, milk powder for middle-aged and elderly people, or nutritional or dietary supplements, etc.

[0134] <Third aspect>

[0135] This invention provides the use of a composition containing two essential components, human milk oligosaccharides and casein phosphopeptides, in the preparation of a nutritional composition that promotes the development of the central nervous system and / or helps improve memory.

[0136] The present invention also provides the use of a composition containing two essential components, human milk oligosaccharides and casein phosphopeptides, in the preparation of foods that promote the development of the central nervous system and / or help improve memory.

[0137] In some specific embodiments, the nutritional composition or food can promote the development of the central nervous system and / or help improve memory.

[0138] In some specific embodiments, the central nervous system development includes brain development; in some preferred embodiments, the central nervous system development includes promoting at least one of neuronal maturation, synapsis, and myelination.

[0139] (Development of the central nervous system)

[0140] Myelination and neuronal maturation

[0141] In some optional embodiments, the myelination and / or neuronal maturation includes promoting the proliferation of oligodendrocyte precursor cells, promoting the myelination of oligodendrocyte precursor cells, and promoting the maturation of oligodendrocyte precursor cells.

[0142] proliferation

[0143] In some specific implementations, promoting the proliferation of oligodendrocyte precursor cells includes increasing the number of cells expressing tetrasialic acid ganglioside (GO1c).

[0144] Myelination and myelination

[0145] In some embodiments, the compositions of the present invention can promote myelination and myelin formation of oligodendrocyte precursor cells, the myelin including myelin in the hippocampus and / or cortex.

[0146] In some specific implementations, promoting myelination of oligodendrocyte precursor cells includes increasing the number of cells expressing myelin-associated glycoprotein (MAG).

[0147] In some exemplary embodiments, the present invention uses magnetic resonance imaging (MRI) and diffusion tensor imaging (DTI) to detect fractional anisotropy (FA), where changes in FA values ​​are closely related to myelin formation.

[0148] In some specific embodiments, the compositions of the present invention significantly increased the FA values ​​of the left and right hippocampi, demonstrating an effect of promoting myelin formation.

[0149] neuronal maturation

[0150] In some embodiments, the compositions of the present invention can promote the further maturation and differentiation of oligodendrocyte progenitor cells (OPCs) into oligodendrocytes (OLs).

[0151] In some specific implementations, promoting the maturation of oligodendrocyte precursor cells includes increasing the number of cells expressing myelin-binding protein (MBP).

[0152] In some specific implementations, promoting neuronal maturation includes promoting the expression of pathway-related proteins in rat brain tissue, such as myelin-binding protein (MBP).

[0153] In some specific embodiments, the present invention provides evidence that the compositions of the present invention can increase the content of myelin in the hippocampus and cortex by detecting longitudinal relaxation rate (R1 = 1 / T1) using magnetic resonance imaging (MRI).

[0154] Synapse

[0155] In some specific implementations, promoting synapsis includes promoting the expression of synapse-related proteins, such as synaptophysin (SYN), postsynaptic density protein 95 (PSD 95), brain-derived neurotrophic factor (BDNF), and growth-associated protein-43 (GAP-43).

[0156] In some exemplary embodiments, the present invention uses magnetic resonance imaging (MRI) and diffusion tensor imaging (DTI) to detect λ1, λ2, and λ3, where λ1 is related to the axial diffusion coefficient and the average diffusion coefficient, and λ2 and λ3 reflect the lateral diffusion coefficients, confirming that the composition of the present invention can increase λ1, that is, can promote axon density in the hippocampus and cortex.

[0157] In some specific embodiments, the compositions of the present invention can significantly promote an increase in axon density in the left and right hippocampi.

[0158] (Aids in improving memory)

[0159] In some specific implementations, the aid to improve memory includes increasing the level of the metabolite gamma-glutamyl in brain tissue. The metabolite gamma-glutamyl can promote the formation of neurite buds and synapses, thereby playing an aid to improve memory.

[0160] In some implementations, the specific selection and ratio of the human milk oligosaccharides and casein phosphopeptides are as described in the preceding <First Aspect>.

[0161] Example

[0162] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0163] Example 1: Cellular Experiment on the Brain-Boosting Effect of the Composition

[0164] Current research on brain development is limited to passive memory studies using animal experiments, without exploring the development of neurons or the effects of specific dosages of the composition on early brain development. This invention uses an in vitro model of primary cell culture containing neurons and OPCs to evaluate the effects of a nutrient mixture on myelin formation and neurons. By experimentally studying the effects of components containing different proportions of 3'-SL, 6'-SL, and casein phosphopeptide on brain nerve cell development, this method systematically and intuitively evaluates the impact on early brain development, and has profound significance for guiding the application of the composition.

[0165] In this invention, an in vitro model of primary cell culture containing neurons and OPCs was used to evaluate the effect of a human milk oligosaccharide composition on myelination. We aimed to add co-nutrients to the mixed cell culture to promote the proliferation, maturation, and differentiation of OPCs into mature OLs and / or the myelination properties of OLs. We first assessed the density of OPCs after 12 days of in vitro culture. Then, this invention assessed the differentiation of OPCs into OLs and the maturation of OLs by quantifying MAG-positive and MBP-positive cells at 18 and 30 days, respectively.

[0166] I. Materials and Methods

[0167] (I) Instruments, Reagents and Consumables

[0168] Cell culture incubator; laser confocal fluorescence microscope; centrifuge; electronic balance; vortex mixer; water bath; tissue homogenizer.

[0169] Nerve cell culture medium; antibodies such as A2B5, MAG, and MBP.

[0170] Reagents:

[0171] 6'-SL: DSM, GLYCARE6SL9001, (Item No.: 5016570);

[0172] 3'-SL: DSM, GLYCARE6SL9001, (Item No.: 5016567);

[0173] Casein phosphopeptide: Guangzhou Green Extract Biotechnology Co., Ltd. (Production batch number: 20240522);

[0174] Olesoxime (Selleck).

[0175] (II) Experimental Methods

[0176] 1. Obtaining primary neural cells

[0177] All experiments were ethically approved. Primary co-culture of neurons and oligodendrocytes (OLs) was performed according to the following steps. In short, the forebrain of newborn fetal rats (SD rats) was dissociated at 37°C for 20 minutes using trypsin (EDT1X, PAN BIOTECH). The reaction was stopped after adding Dulbecco modified Eagle medium (DMEM, PAN BIOTECH) containing DNase Grade II I (0.1 mg / ml, PAN BIOTECH) and 10% fetal bovine serum (FCS, GIBCO). Cells were mechanically separated three times using 10 ml pipettes and then centrifuged at 4°C and 515 g for 10 minutes.

[0178] Live cells were seeded into 96-well plates (20,000 cells / well), pre-coated with poly-L-lysine (BD Falcon) and laminin (Sigma). The culture medium consisted of Neurobasal (GIBCO) supplemented with 2% B27 (GIBCO), 2 mM L-glutamine (L Glu, PAN BIOTECH), 2% P / S solution (PAN BIOTECH), 1% FCS, and 10 ng / ml platelet-derived growth factor (PDGF-AA, PAN BIOTECH). The 96-well plates were stored in a humidified incubator at 37°C in an environment of 95% air and 5% CO2.

[0179] 2. Nerve cell culture

[0180] The same number of cells were placed in 48-well plates and incubated in vitro for 12, 18, or 30 days (Days In Vitro, DIV), with half of the medium replaced every other day, and the mixture or individual nutrients (the above-prepared nutrient composition containing 3'-SL, 6'-SL, and / or casein phosphopeptides) were added to the fresh medium.

[0181] 3. Immunohistochemical experiment

[0182] After 12, 18, and 30 DIV, cells were fixed by treating with a cold mixture of 95% (v / v) ethanol and 5% (v / v) acetic acid for 5 minutes. Then, nonspecific sites were blocked for 15 minutes at room temperature with phosphate-buffered saline (PBS) containing 0.1% (w / w) saponin (Sigma) and 1% (v / v) FCS (GIBCO).

[0183] At 12 DIV, nerve cells were co-incubated with mouse monoclonal antibody A2B5 (dilution: 1 / 200, Millipore, MAB312RX) at room temperature for 2 h, followed by co-incubation with neurofilament protein antibody (dilution: 1 / 500, Sigma, N4142) at room temperature for 2 h. Finally, they were co-incubated with goat anti-rabbit antibody (dilution: 1 / 400, SIGMA, SAB4600084) at room temperature for 1 h.

[0184] At 18 DIV, nerve cells were co-incubated for 2 h with mouse monoclonal antibody MAG (dilution: 1 / 400, Millipore, MAB1567) and neurofilament antibody (dilution: 1 / 500, Sigma, N4142). Then, they were incubated at room temperature for 1 h with secondary antibodies: goat anti-rabbit antibody (dilution: 1 / 400, Sigma, SAB4600042) and goat anti-rabbit antibody (dilution: 1 / 400, SIGMA, SAB4600084).

[0185] At 30 DIV, nerve cells were co-incubated for 2 h with mouse monoclonal antibody MBP (dilution: 1 / 1000, Novus, NBP1-05204) and neurofilament antibody (dilution: 1 / 500, Sigma, N4142). Then, they were co-incubated at room temperature for 1 h with secondary antibodies: goat anti-mouse antibody (dilution: 1 / 800, Sigma, SAB4600042) and goat anti-rabbit antibody (dilution: 1 / 400, SIGMA, SAB4600084).

[0186] 4. Microscopic photography

[0187] 20x magnification was achieved using ImageXpress, equipped with LED lights (360 / 480 / 565 excitation and 460 / 535 / 620 emission). All images were acquired using the same settings.

[0188] Under 12 DIV conditions, the number of OPCs was calculated by quantifying the number of A2B5-expressing cells, and the result is expressed as the average number of A2B5-expressing cells per well per image.

[0189] The differentiation of OPCs into OLs was assessed by counting the number of MAG-positive cells in cell culture. Results are expressed as per image and average cell count per well.

[0190] At 30 DIV, the maturity of OLs was estimated by calculating the number of MBP-positive cells (average number of cells per well per image).

[0191] (III) Grouping of Cell Experiments

[0192] Cells were seeded in 96-well plates and cultured for a certain period of time. Half of the culture medium was replaced every other day. Different concentrations of the mixed or individual test substances were added to fresh primary cell culture medium (added at 12, 18, and 30 days respectively), with 6 replicates for each sample. Olesoxime (300 nM, which has been shown to accelerate OL maturation and myelination in vitro and in vivo, reference: Magalon K, et al. Olesoxime accelerates myelination and promotes repair in models of demyelination. Ann Neurol. 2012; 71(2):213-26.) was used as a positive control. The blank control group, positive control group, and sample intervention group were compared. Immunohistochemistry (MBP, NF, A2B5) was used to determine the effects of casein phosphopeptides alone or in combination and 3'-SL, 6'-SL on OPC population, OL maturation and differentiation, myelin formation, and neurite growth.

[0193] (IV) Statistical Analysis

[0194] Results are expressed as mean ± standard error. SPSS software was used to perform T-tests and one-way ANOVA. A p-value < 0.05 was considered statistically significant.

[0195] II. Experimental Results

[0196] 1. Effects of each sample on nerve cells

[0197] To measure the effect of mixed or individual nutrient treatments on OPCs, we assessed the number of A2B5-labeled positive cells after 12 DIV to estimate the number of OPCs.

[0198] Figure 1A The effects of different nutrient compositions on nerve cells cultured in vitro for 12 days are shown in the A2B5 immunostaining pattern (green). Scale bar: 50 μm. The measured values ​​for each image and the average number of A2B5-positive cells per well are shown in Table 1.

[0199] Table 1 shows the effects of 3'-SL and 6'-SL nutrient composition samples on the number of A2B5 positive cells.

[0200]

[0201]

[0202] Sample treatment results showed that, compared with the blank control group, 3'-SL and 6'-SL significantly increased the number of A2B5 positive cells, indicating that both components contribute to the proliferation of oligodendrocyte precursor cells. However, the inventors discovered that a significant synergistic effect was observed when the two components were used in combination at certain ratios. Sample treatment results showed that, compared with the blank control group (cell experiment group 1), cell experiment groups 9 and 10 showed that a synergistic promotion of oligodendrocyte precursor cell proliferation occurred when the ratio of 3'-SL to 6'-SL was 1:2 and 5:2.

[0203] In terms of components and dosages, cell experiment group 9 is equivalent to a combination of cell experiment groups 3 and 5. Compared to the blank control group (cell experiment group 1), cell experiment groups 3 and 5 increased the number of A2B5 positive cells by 26.33 and 23, respectively, while cell experiment group 9 increased the number of A2B5 positive cells by 59, which is not only greater than the sum of the former two (49.33), but also greater than each of the former two individually. The same conclusion can be drawn for cell experiment group 10.

[0204] Subsequently, the inventors further added casein phosphopeptides to the composition to culture nerve cells for the next stage of the experiment. Figure 1B The effects of different nutrient compositions on nerve cells cultured in vitro for 12 days are shown in the A2B5 immunostaining images (green). Scale bar: 50 μm. The measured values ​​for each image and the average number of A2B5-positive cells per well are shown in Table 2.

[0205] Table 2 shows the effect of different nutrient composition samples containing 3'-SL, 6'-SL and / or casein phosphopeptides on the number of A2B5 positive cells.

[0206]

[0207]

[0208] Sample processing results showed that, compared with the blank control group, the combination of 3'-SL, 6'-SL, and casein phosphopeptide significantly increased the number of A2B5 positive cells, indicating that all three components contribute to the proliferation of oligodendrocyte precursor cells. However, the inventors discovered that when 3'-SL, 6'-SL, and casein phosphopeptide are mixed in a certain proportion, they synergistically increase the number of A2B5 positive cells and thus promote the proliferation of oligodendrocyte precursor cells. Specifically, this can be seen from cell experimental groups 10 and 11:

[0209] In terms of the components and dosages used, cell group 11 is equivalent to a combination of cell experimental groups 4, 6, and 9. Compared to the blank control group (cell experimental group 1), cell experimental groups 4, 6, and 9 increased the number of A2B5 positive cells by 45.33, 46.33, and 58.33, respectively, while cell experimental group 10 increased the number of A2B5 positive cells by 186.33, which is not only much greater than the sum of the previous three (149.99) (the difference Δ1 = 36.34 from the sum of the previous two), but also greater than each of the previous three.

[0210] Cell experiment group 11 is equivalent to a combination of cell experiment groups 3, 5, and 9. However, compared to the blank control group (cell experiment group 1), cell experiment groups 3, 5, and 9 increased the number of A2B5 positive cells by 7, 11.33, and 58.33, respectively, while cell experiment group 11 increased the number of A2B5 positive cells by 148.33. This increase is not only much greater than the sum of the previous three groups (76.66) (the difference Δ2 = 71.67), but also greater than each of the previous three groups individually.

[0211] This indicates that when 3'-SL, 6'-SL, and casein phosphopeptide are used in combination in a certain proportion, there is a synergistic effect among the three components, which can synergistically increase the number of A2B5 positive cells and promote the proliferation of oligodendrocyte precursor cells.

[0212] 2. Effects of each sample on myelination of nerve cells

[0213] To measure the effect of mixed or individual nutrient treatments on myelination of OPCs, we assessed the number of MAG-positive cells after 18 DIV.

[0214] Figure 2 The effects of different nutrient compositions on nerve cells cultured in vitro for 18 days are shown in the MAG immunostaining images (green). Scale bar: 50 μm. The measured values ​​for each image and the average number of MAG-positive cells per well are shown in Table 3.

[0215] Table 3 shows the effects of 3'-SL, 6'-SL, and casein phosphopeptide nutrient composition samples on the number of MAG-positive cells.

[0216]

[0217]

[0218] Sample processing results showed that, compared with the blank control group, 3'-SL and 6'-SL significantly increased the number of MAG-positive cells, indicating that both components contribute to the proliferation of oligodendrocyte precursor cells. However, the inventors discovered that a significant synergistic effect was observed when the two components were used in combination at a certain ratio. Sample processing results showed that, compared with the blank control group (cell experiment group 1), cell experiment groups 7 and 8 showed that a 1:2 ratio of 3'-SL to 6'-SL synergistically increased the number of MAG-positive cells and promoted myelination of oligodendrocyte precursor cells.

[0219] In terms of the components and dosages used, cell experiment group 7 is equivalent to a combination of cell experiment groups 4 and 6. Compared to the blank control group (cell experiment group 1), cell experiment groups 4 and 6 increased the number of MAG-positive cells by 27.33 and 42, respectively, while cell experiment group 7 increased the number of MAG-positive cells by 74.33, which is not only greater than the sum of the former two (69.33), but also greater than each of the former two individually.

[0220] Cell experiment 8 is equivalent to a combination of cell experiment groups 3 and 5. Compared to the blank control group (cell experiment group 1), cell experiment groups 3 and 5 increased the number of MAG-positive cells by 7.67 and 21.67, respectively, while cell experiment group 8 increased the number of MAG-positive cells by 41.67, which is not only greater than the sum of the former two (29.34), but also greater than each of the former two individually.

[0221] The inventors further discovered that when 3'-SL, 6'-SL, and casein phosphopeptide are combined, they can synergistically promote myelination of oligodendrocyte precursor cells. In particular, this can be observed in cell experimental groups 13-16.

[0222] In terms of the components and dosages used, cell experiment group 13 is equivalent to a combination of cell experiment groups 4, 6, and 11. Compared to the blank control group (cell experiment group 1), cell experiment groups 4, 6, and 11 increased the number of MAG-positive cells by 27.33, 42, and 40.33, respectively, while cell experiment group 13 increased the number of MAG-positive cells by 170.33, which is not only much greater than the sum of the previous three (109.66) (the difference from the sum of the previous three is Δ1 = 60.67), but also greater than each of the previous three.

[0223] Cell experiment group 15 is equivalent to a combination of cell experiment groups 3, 5, and 11. However, compared to the blank control group (cell experiment group 1), cell experiment groups 3, 5, and 11 increased the number of MAG-positive cells by 7.67, 21.67, and 40.33, respectively, while cell experiment group 15 increased the number of MAG-positive cells by 144.67, which is not only much greater than the sum of the previous three (69.67) (the difference from the sum of the previous three is Δ2 = 75), but also greater than each of the previous three.

[0224] Similarly, this conclusion can be drawn for cell experiment groups 14 and 16.

[0225] This indicates that when 3'-SL, 6'-SL, and casein phosphopeptide are used in combination in a certain proportion, there is a synergistic effect among the three components, which can synergistically increase the number of MAG-positive cells and promote myelination of oligodendrocyte precursor cells.

[0226] Oligodendrocyte precursor cells (OPCs) are the main glial cell population in the central nervous system, accounting for 2%-9% of the total cell population. OPCs specifically express tetrasialic ganglioside (GOlc) on their surface, which can be recognized by the monoclonal antibody A2B5. After mitosis, OPCs differentiate into myelinated oligodendrocytes (OLs), highly specialized cells in the central nervous system (CNS). Their unique characteristic lies in the production of myelin, a multilayered, lipid-rich sheath that covers and insulates neuronal axons, increasing the speed of electrical signal transmission and providing metabolic support for neurons. Furthermore, the myelin sheath provides essential metabolic support for axonal function, promoting oxidative phosphorylation of axonal mitochondria. Myelinated OLs express MAG, and MAG expression gradually increases during OL maturation. MAG is mainly expressed in the peri-axonal region of the myelin sheath.

[0227] 3. Effects of each sample on OPC cell maturation

[0228] To measure the effect of mixed or individual nutrient treatments on OPC cell maturation, we assessed the number of MBP-labeled positive cells after 30 DIV.

[0229] Figure 3 The effect of different nutrient compositions on nerve cells cultured in vitro for 30 days is shown in the MBP immunostaining image (green). Scale bar: 50 μm. The measured values ​​for each image and the average number of MBP-positive cells per well are shown in Table 4.

[0230] Table 4 shows the effects of 3'-SL, 6'-SL, and casein phosphopeptide nutrient composition samples on the number of MBP-positive cells.

[0231]

[0232]

[0233] Sample processing results showed that, compared with the blank control group, 3'-SL and 6'-SL significantly increased the number of MBP-positive cells, indicating that both components contribute to the proliferation of oligodendrocyte precursor cells. However, the inventors discovered that a significant synergistic effect was observed when the two components were used in combination at a certain ratio. Sample processing results showed that, compared with the blank control group (cell experiment group 1), cell experiment group 7 showed that a 1:2 ratio of 3'-SL to 6'-SL synergistically increased the number of MAG-positive cells and promoted myelination of oligodendrocyte precursor cells.

[0234] In terms of the components and dosages used, cell experiment group 7 is equivalent to a combination of cell experiment groups 3 and 5. Compared to the blank control group (cell experiment group 1), cell experiment groups 3 and 5 increased the number of MAG-positive cells by 10.34 and 19.34, respectively, while cell experiment group 7 increased the number of MAG-positive cells by 33, which is not only greater than the sum of the former two (29.68), but also greater than each of the former two individually.

[0235] Further investigation revealed that, compared to the blank control group, the combination of 3'-SL, 6'-SL, and casein phosphopeptide significantly increased the number of MBP-positive cells, indicating that all three components contribute to the proliferation of oligodendrocyte precursor cells. However, the inventors found that a mixture of 3'-SL, 6'-SL, and casein phosphopeptide in a certain proportion increased the number of MBP-positive cells and facilitated the maturation of oligodendrocyte precursor cells.

[0236] Specifically, based on cell experiment groups 10 and 11, it can be seen that:

[0237] In terms of components and dosages, cell experiment group 10 is equivalent to a combination of cell experiment groups 4, 6, and 8. Compared to the blank control group (cell experiment group 1), cell experiment groups 4, 6, and 8 increased the number of MBP-positive cells by 53.34, 43, and 44, respectively, while cell experiment group 10 increased the number of MBP-positive cells by 168, which is not only much greater than the sum of the previous three (140.34), but also greater than each of the previous three.

[0238] Cell experiment group 11 is equivalent to a combination of cell experiment groups 3, 5, and 8. However, compared to the blank control group (cell experiment group 1), cell experiment groups 3, 5, and 8 increased the number of MBP-positive cells by 10.34, 19.34, and 44, respectively, while cell experiment group 11 increased the number of MBP-positive cells by 132.67. This increase is not only much greater than the sum of the previous three groups (73.68) (the difference Δ1 = 58.99), but also greater than each of the previous three groups individually.

[0239] This indicates that when 3'-SL, 6'-SL, and casein phosphopeptide are used in combination in a certain proportion, there is a synergistic effect among the three components, which can synergistically increase the number of MBP-positive cells and help the maturation of oligodendrocyte precursor cells.

[0240] Example 2: Animal experiments on the brain-boosting effects of the composition

[0241] I. Materials and Methods

[0242] (I) Instruments, Reagents and Consumables

[0243] Instruments: Electronic balance (FA1004B) Shanghai Precision Instruments Co., Ltd., Benchtop high-speed centrifuge (TG16-WS) Xiangyi Centrifuge Instrument Co., Ltd., Multifunctional microplate reader SynergyHTX BioTek, Ultra-low temperature freezer (DW-86L338J) Qingdao Haier Biomedical Co., Ltd., Protein electrophoresis system PowerPac HC Bio-Rad, Electroporation membrane transfer device Trans-Blot SD Bio-Rad, Microchemi gel imaging system DNR Bioimaging Systems Co., Ltd., Small animal magnetic resonance imaging equipment 7.0T PharmaScan 70T / 16USBruker.

[0244] Reagents: BCA protein quantification kit (Beyotime Biotechnology Co., Ltd.); PVDF membrane (Amersham Biotechnology Co., Ltd., USA); ECL ultrasensitive chemiluminescence developing solution (Beyotime Biotechnology Co., Ltd.); Antibodies A2B5 (Invitrogen), BDNF (Proteintech), GAP43 (Proteintech), MAG (Millipore), MBP (Novus), PLP (Proteintech), PSD95 (Proteintech), SYN (Proteintech), GAPDH (UtiBody).

[0245] (II) Experimental Methods

[0246] 1. Grouping and intervention of experimental animals

[0247] Three-week-old male SD rats (n=66) were acclimatized to the diet for one week and then randomly divided into 6 groups (n=11), as shown in Table 5. During the experiment, all rats were fed a growth and reproduction diet. The control group was administered physiological saline by gavage at 1 mL / 100 g / day, while the other groups were administered the corresponding nutrients by gavage. Food intake and body weight were measured weekly during the intervention period. After the fourth week of intervention, the rats were fasted for 12 hours, anesthetized by intraperitoneal injection of ketamine (100 mg / kg·bw), and blood was collected from the eyeballs. Serum was obtained by centrifuging the rat blood at 4℃, 3000 rpm for 15 min and stored at -80℃.

[0248] Table 5. Test Dosage and Group

[0249]

[0250]

[0251] Note: "—" indicates that no content was added or no proportional relationship exists.

[0252] 2. Body weight measurement

[0253] Body weight and weekly food intake of SD rats were measured on days 0, 7, 14, 21, and 28.

[0254] (1) The ratio of animal organs and body fat content

[0255] After euthanasia, organs and tissues were harvested and weighed to calculate the organ index and body fat percentage. Organ index = organ weight (g) / 100g rat body weight.

[0256] (2) Detection of synapse-related proteins and myelin-related proteins in brain tissue by WES method

[0257] Rat brain tissue was collected, lysed, and total protein was extracted. Using GAPDH as an internal control, Western blot analysis was performed to determine the levels of synapse-related proteins in the brain tissue, including synaptophysin (Syn), postsynaptic density protein-95 (PSD)-95, brain-derived neurotrophic factor (BDNF), and growth-associated protein-43 (GAP-43). Myelin-related markers included myelin PLP (PLP), myelin basic protein (MBP), oligodendrocyte-specific tetrasialotetrahexosylganglioside (recognized using monoclonal antibody A2B5), and MAG protein.

[0258] 3. Magnetic resonance imaging of rat brain tissue

[0259] (1) Magnetic resonance imaging (MRI) scan: High-resolution T1-weighted structural images were used to observe the hippocampus and cortex; (2) Magnetic resonance diffusion tensor imaging (DTI) scan: The hippocampus and cortex were observed; (3) Magnetic resonance spectroscopy (MRS) was used to detect and analyze metabolic changes in the hippocampus.

[0260] 4. Statistical Analysis

[0261] Results are expressed as mean ± standard error (mean ± SEM). SPSS software was used to perform T-test and one-way ANOVA. A p < 0.05 was considered statistically significant, and a p < 0.01 was considered highly statistically significant.

[0262] II. Experimental Results

[0263] 1. Effects of the composition on rat body weight, food intake, organ index, and body fat percentage

[0264] The effects of the composition on rat body weight, food intake, organ index and body fat percentage are shown in Tables 6-8 below.

[0265] Table 6 Effects of the composition on rat body weight (g)

[0266]

[0267]

[0268] During the first two weeks of intervention, the body weight of rats in each group increased rapidly, with no significant difference among the groups. In the third week of intervention, the rate of weight gain decreased, and growth began to slow down; by the fourth week, the rate of weight gain in each group was the slowest. At the end of the feeding period, the body weight of the control group rats was 324.63±12.81g, and the body weight of the six groups of rats was slightly higher than that of the control group, but the difference was not significant.

[0269] Table 7 Effects of the composition on food intake in rats

[0270]

[0271] Throughout the experiment, the food intake of rats in each group showed a trend of first increasing and then decreasing. Food intake was higher in the first two weeks and gradually decreased in the following two weeks, consistent with the rate of weight gain. There was no significant difference in food intake among the intervention groups compared to the control group.

[0272] Table 8 Effects of the composition on organ index and body fat percentage in rats

[0273]

[0274]

[0275] The organ indices (heart, liver, spleen, and kidneys) of each group showed no significant difference compared to the control group, indicating that the selected composition did not affect the organ indices of normal rats. Compared to the control group, the body fat percentage of each intervention group remained unchanged, indicating that the selected composition did not significantly affect the body fat percentage of normal rats.

[0276] 2. Expression of pathway-related proteins in the brain tissue of rats in each group

[0277] The expression of pathway-related proteins in the brain tissue of rats in each group is shown in Tables 9 and 9-1 below.

[0278] Table 9. Effects of the composition on synaptic and myelin-related proteins in rat brain tissue.

[0279]

[0280] Note: "*" indicates that compared with the blank control group, p < 0.05; "**" indicates that compared with the blank control group, p < 0.01.

[0281] Table 9-1 Effects of the composition on synaptic and myelin-related proteins in rat brain tissue

[0282] Animal experimental group Dosage group PLP A2B5 MAG 1 Blank control group 0.20±0.02 0.20±0.02 0.21±0.01 5 3'-SL+6'-SL+CPP high <![CDATA[0.26±0.02 * ]]> <![CDATA[0.26±0.02 * ]]> <![CDATA[0.27±0.02 ** ]]> 6 3'-SL+6'-SL+CPP low <![CDATA[0.29±0.03 ** ]]> <![CDATA[0.30±0.03 ** ]]> <![CDATA[0.30±0.02 ** ]]>

[0283] Synaptophysin (SYP or Syn), also known as synaptic vesicle protein, accounts for approximately 8% of total vesicle protein and is an acidic calcium-binding protein specifically expressed on the vesicle membrane. When a nerve is excited, SYP is transported to the presynaptic vesicle membrane at the axon terminal, driving its carboxyl terminus to bind with calcium. 2+ On the other hand, it enhances the phosphorylation of tyrosine protein kinases, promoting the fusion of synaptic vesicles with the plasma membrane, thereby releasing neurotransmitters into the interstitial space. Therefore, the location and density of SYP expression can directly reflect the number of vesicles and indirectly reflect the number and localization of synapses, which affects neurotransmitter release and thus regulates signal transmission efficiency. This is considered one of the key mechanisms affecting long-term potentiation (LTP) synaptic efficacy.

[0284] Synapses are the sites of strongest neural remodeling, and many functions of the nervous system depend on the participation of synaptic plasticity. Synaptophysin and growth-associated protein-43 (GAP-43) are proteins closely related to synaptic plasticity, and their expression levels are closely related to synaptic structural plasticity.

[0285] The postsynaptic density protein (PSD) is a semi-circular, band-shaped active region located at the junction of the postsynaptic membrane and cytoplasm. Composed of various skeletal and regulatory proteins, it forms a crucial morphological basis for synaptic structure. Among these, postsynaptic density protein 95 (PSD-95) is the most densely expressed synaptic scaffold protein within the PSD. It interacts with NMDAR and isoxazolylpropionate receptors, stabilizing and transporting these proteins to the postsynaptic membrane, contributing to axon formation and LTP induction, and serving as a marker of excitatory postsynaptic density. Furthermore, the binding of PSD-95 to NMDAR transmits signals into the cell, such as the retrograde messenger nitric oxide (NO), catalyzed by neuronal nitric oxide synthase (nNOS), which diffuses back to the presynaptic neuron, promoting neurotransmitter release through a series of biological reactions. Therefore, the distribution and expression of synaptic plasticity markers SYP and PSD-95 directly reflect the degree and variability of LTP induction.

[0286] Brain-derived neurotrophic factor (BDNF) is one of the most studied neurotrophic factors, playing a role in regulating neurite growth and branching in neurons. BDNF binding to its contained CRE sequence can rapidly enhance transcriptional efficiency, exerting biological effects related to long-term neurotrophic factors (LTPs).

[0287] Sample processing results showed that, compared with the blank control group, the high-dose casein phosphopeptide group, when used alone within the tested dose range, significantly increased the expression of synaptic-associated proteins (SYP, PSD, GAP-43, and BDNF). The 3'-SL+6'-SL combination itself did not promote the expression of synaptic-associated proteins, but when used in combination with casein phosphopeptide, it significantly increased the promoting effect of casein phosphopeptide on the expression of synaptic-associated proteins. This indicates that the combination of 3'-SL+6'-SL and casein phosphopeptide has a synergistic effect and can synergistically promote the expression of synaptic-associated proteins.

[0288] Oligodendrocyte precursor cells (OPCs) are the main glial cell population in the central nervous system, accounting for 2%-9% of the total cell population. A2B5 is a cell surface marker of OPCs. After mitosis, OPCs differentiate into myelinated oligodendrocytes (OLs), which are highly specialized cells in the central nervous system (CNS). Their unique characteristic is the production of myelin, a multilayered, lipid-rich sheath that covers and insulates neuronal axons, increasing the speed of electrical signal transmission and providing metabolic support for neurons. Furthermore, the myelin sheath provides essential metabolic support for axonal function and promotes oxidative phosphorylation of axonal mitochondria. Myelinated OLs express MAG, and MAG expression gradually increases during OL maturation. MAG is mainly expressed in the peri-axonal region of the myelin sheath. Fully mature OLs generate myelin-associated proteins, such as myelin basic protein (MBP) and myelin proteolipid protein (PLP). Subsequently, OLs expand, interact with neuronal axons, and encapsulate them. MBP, a crucial protein in myelination, is responsible for intracellular myelin compaction, functioning by binding the cytoplasmic surface of the myelin membrane. Sample treatment results showed that, compared to the blank control group, the combination of 3'-SL+6'-SL and casein phosphopeptide significantly increased the expression of myelin-related proteins in all groups. Particularly in MBP protein expression, the 3'-SL+6'-SL composition and casein phosphopeptide exhibited a synergistic effect, jointly promoting MBP protein expression. This indicates that the use of the 3'-SL+6'-SL and casein phosphopeptide combination helps to synergistically promote myelin compaction in animals.

[0289] 3. Effects of the composition on magnetic resonance imaging results of rat brain tissue

[0290] Myelin is a fatty substance that forms a protective layer around axons, enhancing their conductivity. In vertebrates, axons are encased in a myelin sheath to ensure rapid transmission of information between neurons as electrical signals within the central nervous system. The myelin sheath consists of repetitive units of a tight oligodendrocyte membrane, a bilayer separated by a water layer and a 3-4 nm extracellular space. Unlike membranes in other cell types, myelin is typically characterized by a lipid and protein weight composition of 50%, with the myelin membrane consisting of 75%-80% lipids and 20%-25% protein. Increased myelin content is associated with longitudinal relaxation rate (R1 = 1 / T1) on magnetic resonance imaging (MRI). Mature white matter is characterized by increased myelin content, leading to an increased longitudinal relaxation rate (R1) due to the interaction of free water molecules with macromolecules (where R1 = 1 / T1, and the relaxation time T1 is shortened). Therefore, T1 localization is a sensitive quantitative method for myelin formation and has been studied in premature infants, normal infants, and children and adolescents. Figure 4 ).

[0291] Table 10 Effects of the composition on T1 in rat brain tissue

[0292]

[0293] Note: "*" indicates that compared with the blank control group, p < 0.05; "**" indicates that compared with the blank control group, p < 0.01.

[0294] Table 10 shows that the combination of 3'-SL+6'-SL and casein phosphopeptide (groups 5 and 6) significantly increased the content of myelin in the hippocampus and left cortex. 3'-SL+6'-SL alone did not show a significant effect, but when combined with casein phosphopeptide, it showed a significant effect compared to the control group, indicating that the combination of groups 5 and 6 has a synergistic effect.

[0295] Magnetic resonance imaging (MRI) allows for the quantification of diffusion-based microstructural integrity. The most commonly used method is diffusion tensor imaging (DTI), which models the diffusion coefficient of water as isotropic, free-moving, and anisotropic diffusion coefficients. DTI serves a dual purpose. First, based on diffusion coefficient measurements along multiple directions, virtual reconstruction of fiber bundles can be achieved. Second, the diffusion characteristics of individual voxels can be quantified using different metrics. In turn, these metrics represent estimates of the microstructural integrity within a given voxel. The most widely used metric is fractional anisotropy (FA), which represents the degree of directional diffusion of water. Axon diameter or axon density can be reflected by FA, ​​and there is a positive correlation between FA and fiber density in white matter regions. Changes in FA values ​​are closely related to myelin sheath formation; increased myelin sheathing inhibits water molecule diffusion, manifesting as an increase in FA values. λ1 is related to the axial diffusion coefficient and the average diffusion coefficient, while λ2 and λ3 reflect the lateral diffusion coefficient. Figure 5 ).

[0296] Table 11 Effects of the composition on DTI in the right hippocampus of rats

[0297]

[0298] Table 11-1 Effects of the composition on DTI in the right hippocampus of rats

[0299] Animal experimental group Dosage group FA λ1 1 Blank control group 58.13±6.88 110.33±9.95 2 3'-SL+6'-SL 65.21±4.23 106.10±9.68 3 Low dose of casein phosphopeptide 56.07±6.50 114.00±9.17 6 3'-SL+6'-SL+CPP low 79.17±6.24** 123.00±5.27

[0300] Note: "**" indicates a comparison with the blank control group, p < 0.01.

[0301] Table 12 Effects of the composition on DTI in the left hippocampus of rats

[0302]

[0303] Table 12-1 Effects of the composition on DTI in the left hippocampus of rats

[0304]

[0305]

[0306] Note: "**" indicates a comparison with the blank control group, p < 0.01.

[0307] Table 13 Effects of the composition on DTI in the right cortex of rats

[0308]

[0309] Table 13-1 Effects of the composition on DTI in the right cortex of rats

[0310] Animal experimental group Dosage group FA λ1 1 Blank control group 64.77±24.08 120.33±17.67 2 3'-SL+6'-SL 63.83±14.36 118.67±4.73 3 Low dose of casein phosphopeptide 61.47±13.91 116.93±11.64 6 3'-SL+6'-SL+CPP low 71.37±15.96 125.33±10.69

[0311] Table 14 Effects of the composition on left cortical DTI in rats

[0312]

[0313] Table 14-1 Effects of the composition on left cortical DTI in rats

[0314]

[0315]

[0316] As shown in Tables 11 to 14, group 5 increased the FA and λ1 values ​​in the hippocampus and cortex, demonstrating its role in promoting myelin formation and increasing axon density.

[0317] Tables 11-1 to 14-1 show that group 6 increased the FA and λ1 values ​​in the hippocampus and also exhibited a promoting effect. In the left hippocampus, group 6 significantly promoted myelin formation and increased axon density. The combination of groups 6 showed a synergistic effect in promoting myelin formation and increasing axon density in the left hippocampus. In the right hippocampus, group 6 significantly increased the FA value, demonstrating a significant promoting effect on myelin formation. Group 5 also showed a promoting effect. The results indicate that the combination of groups 6 has a synergistic effect in promoting myelin formation in the right hippocampus.

[0318] Table 15 Effects of the composition on MRS in rat brain tissue

[0319]

[0320] Note: "*" indicates a comparison with the blank control group, p < 0.05; "**" indicates a comparison with the blank control group, p < 0.01.

[0321] Magnetic resonance spectroscopy (MRS) is currently the only technique that can non-invasively observe metabolic and biochemical changes in living tissues. Metabolites of interest in MRS studies include N-acetylaspartic acid, choline, lipids, lactate, gamma-glutamyl, inositol, and taurine. Table 15 shows that the composition had no significant effect on the levels of N-acetylaspartic acid, inositol, taurine, choline, and lactate, and the composition did not affect normal brain tissue metabolism.

[0322] Glutamine (GLN) is a precursor to the neurotransmitters glutamate, aspartate (ASP), and the inhibitory amino acid GABA, thus GLN levels play a crucial role in neurotransmission. Glutamate (GLU) is one of the main excitatory neurotransmitters in the brain and may play a role in brain development by influencing neurite budding, synapsis, and dendritic pruning. Glutamate (GLU) is responsible for transmitting signals between nerve cells and plays an important role in learning and memory under normal conditions. The glutamate and glutamine pool (Glx) is a marker of the glutamate-glutamine cycle and is believed to function between neurons and astrocytes, playing a key role in the glutamate / γ-aminobutyric acid (GABA)-glutamine metabolic pathway, which is essential for normal brain cell function. As shown in Table 15, compared with the control group, groups 3, 4, 5, and 6 showed significantly increased Glx levels, suggesting that a certain concentration of casein phosphopeptide may promote the increase of Glx in the brain. Because group 2 (oligosaccharides) did not significantly increase Glx, and group 4 (casein phosphopeptides) increased less than group 6, oligosaccharides and casein phosphopeptides in group 6 have a synergistic effect.

[0323] In summary, this invention provides insights into the application of a combination of 3'-SL, 6'-SL, and casein phosphopeptides in brain development, particularly in neural development, offering new avenues for the development of future functional foods. 3'-SL, 6'-SL, and casein phosphopeptides show great promise in improving memory and brain development. Studies have revealed that the extracellular environment plays a crucial role in regulating brain homeostasis and controlling myelination during central nervous system development. Deficiencies in key nutrients significantly impact brain development. Our research demonstrates that, in in vitro models, brain cell cultures treated with the combination of 3'-SL, 6'-SL, and casein phosphopeptides increased the number of OPCs, promoted differentiation or maturation into OLs, and enhanced the myelination properties of OLs, exhibiting a good synergistic effect among the components. Animal experiments further corroborate the conclusions of our cell experiments.

[0324] It should be noted that although the technical solution of the present invention has been described with specific examples, those skilled in the art will understand that the present invention should not be limited thereto.

[0325] The various embodiments of the present invention have been described above. These descriptions are exemplary and not exhaustive, nor are they limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is chosen to best explain the principles, practical application, or technical improvements to the embodiments in the market, or to enable others skilled in the art to understand the embodiments disclosed herein.

Claims

1. A nutritional composition for promoting central nervous system development, characterized in that, The nutritional composition comprises the following essential components: a human milk oligosaccharide component and a casein phosphopeptide component; and, in the nutritional composition, the mass ratio of the human milk oligosaccharide component to the casein phosphopeptide component is (1-100):(500-1).

2. The nutritional composition according to claim 1, characterized in that, In the nutritional composition, the human milk oligosaccharide component comprises one or more of 3'-sialyllactose, 6'-sialyllactose, 4'-galactosyllactose, 3'-galactosyllactose, 2'-fucosyllactose, lacto-N-tetraose, lactodifucosyltetraose, lacto-N-neotetraose, 6'-galactosyllactose, and lacto-N-fucopentaose-I, Preferably, the human milk oligosaccharide component comprises 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL).

3. The nutritional composition according to claim 2, characterized in that, In the human milk oligosaccharide component, the mass ratio of 3'-sialyllactose (3'-SL) to 6'-sialyllactose (6'-SL) is (1-15):(15-1).

4. The composition according to any one of claims 1 to 3, characterized in that, In the human milk oligosaccharide component, the mass ratio of 3'-sialyllactose (3'-SL) to 6'-sialyllactose (6'-SL) is (1-10):(10-1), preferably (1-5):(5-1).

5. A food product, characterized by, The food comprises or uses the nutritional composition according to any one of claims 1-4.

6. The food product of claim 5, wherein, The food includes any one or more of infant food, child food, adolescent food, pregnant and lying-in woman food, adult food, and middle-aged and elderly food.

7. The food according to claim 5 or 6, characterized in that, The food can be in solid form, semi-solid form, or liquid form.

8. Use of a composition containing both essential components of human milk oligosaccharide and casein phosphopeptide in the preparation of a nutritional composition for promoting central nervous system development and / or assisting in improving memory.

9. Use of a composition containing both essential components of human milk oligosaccharide and casein phosphopeptide in the preparation of a food for promoting central nervous system development and / or assisting in improving memory.

10. The use according to claim 8 or 9, characterized in that, The central nervous system development includes brain development; Preferably, the central nervous system development includes at least one of neuron maturation, synapse genesis, and myelination.

11. Use according to claim 10, characterized in that, The central nervous system development includes promoting the proliferation of oligodendrocyte precursor cells, promoting the myelination of oligodendrocyte precursor cells, and promoting the maturation of oligodendrocyte precursor cells.

12. Use according to claim 10, characterized in that, The central nervous system development includes promoting the expression of synapse-related proteins, preferably the synapse-related proteins include at least one of synaptophysin (SYN), postsynaptic density protein 95 (PSD 95), brain-derived neurotrophic factor (BDNF), and growth associated protein-43 (GAP-43).

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