Brain development composition containing breast milk oligosaccharide and application

Abstract

The invention discloses a brain development composition containing breast milk oligosaccharide and application. The invention provides a brain development composition containing breast milk oligosaccharide. The brain development composition comprises the following necessary components: a breast milk oligosaccharide component, a whey protein component and a phospholipid component, and in the composition, the mass ratio of the breast milk oligosaccharide component to the whey protein component to the phospholipid component is (0.1-15): (500-1): (1000-0.5). The brain development composition provided by the invention can be suitable for promoting brain development and cognitive functions, can remarkably promote brain development by scientifically matching the breast milk oligosaccharide component, the whey protein component and the phospholipid component, and is also beneficial to conveniently adding or preparing various foods or health-care products.

Description

Technical Field

[0001] This invention relates to a brain development composition, belonging to the food field, and more specifically, to a brain development composition containing human milk oligosaccharides and its application. Background Technology

[0002] Brain development continues for some time after birth. During preschool years, the brain quadruples in size, reaching approximately 90% of its adult volume by age six. Structural changes in the major gray and white matter regions persist into childhood and adolescence; these structural changes parallel functional changes and are reflected in behavior. In early infancy, the level of connectivity throughout the developing brain far exceeds that of an adult. This vigorous connectivity is gradually pruned through competitive processes influenced by biological experience. These early experience-dependent processes underlie well-documented plasticity and adaptability, hallmarks of early brain development.

[0003] Mounting evidence suggests that early life nutrition is crucial for the brain to reach its full developmental potential. The effects of early life nutritional support have been demonstrated through preclinical models, showing that nutritional deficiencies coinciding with peak brain growth can have lifelong impacts on brain development.

[0004] 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 neurons (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.

[0005] 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.

[0006] Existing technologies have conducted some research on nutritional compositions that promote brain development, for example:

[0007] 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.

[0008] 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.

[0009] 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.

[0010] 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 brain development, such as those that promote central nervous system development and / or help improve memory.

[0011] 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.

[0012] Phospholipids are a collective term for the various components of phospholipids. They refer to lipids containing phosphoric acid and belong to complex lipids. They are composed of glycerol or sphingosine, fatty acids, phosphoric acid, and nitrogen-containing compounds, and are divided into two main categories: glycerophospholipids and sphingosine. In glycerophospholipids, the two hydroxyl groups of glycerol are esterified by fatty acids to form phosphatidic acid. The phosphate hydroxyl groups of phosphatidic acid are then replaced by amino alcohols or inositol, forming several major glycerophospholipids such as phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylinositol (PI), and phosphatidic acid (PA).

[0013] Alpha-lactalbumin (α-LA) is an active protein with multiple biological functions, providing the body with abundant essential amino acids such as tryptophan and lysine. It is the main whey protein in breast milk, accounting for approximately 22% of total whey protein, and provides energy and nutrition for the physical and brain development of infants.

[0014] However, in existing research, the mechanism of action, actual effects, and degree of efficacy of the combined use of human milk oligosaccharides, whey protein, and phospholipids in promoting neurodevelopment remain unclear. Current research has not yet been able to definitively reveal how these three components synergistically affect brain development processes when present in specific combinations, nor has it clarified their impact on cognitive enhancement.

[0015] References

[0016] Reference 1 CN119949517A

[0017] Reference 2CN118252257A Summary of the Invention

[0018] The problem the invention aims to solve

[0019] 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, synapsis, and other processes that promote brain development 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 topics related to promoting myelin development, neuronal maturation, and synapsis that promote brain development.

[0020] Therefore, there is a need for a food that provides a nutritional composition that can promote neurodevelopment such as myelination and is also absorbable and usable in order to better exert its promoting effect on brain development and cognitive function.

[0021] In view of this, the present invention provides a nutritional composition comprising human milk oligosaccharide components, whey protein components, and phospholipid components, which significantly promotes brain development, including myelin sheath development and formation. Specifically, the present invention explores the effects of the composition on myelin cell development and studies the effects of the composition and its component ratios on early brain development. By scientifically designing the ratio of human milk oligosaccharides, whey protein, and milk phospholipid components in the composition, a synergistic effect can be achieved, meeting market demand for functional compositions. This brain development-promoting composition provided by the present invention offers a safe and effective nutritional composition for promoting healthy brain nerve development, particularly for people of different age groups, thereby enhancing various brain functions such as intelligence.

[0022] Solution for solving the problem

[0023] [1]. A brain development composition containing human milk oligosaccharides, wherein the brain development composition comprises the following essential components: human milk oligosaccharide component, whey protein component and phospholipid component; and, in the brain development composition, the mass ratio of the human milk oligosaccharide component, the whey protein component and the phospholipid component is (0.1-15):(500-1):(1000-0.5).

[0024] [2]. The brain development 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'-fucosyllactose, lactose-N-tetrasaccharide, lactose-difucosyltetrasaccharide, 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).

[0025] [3]. The brain development composition according to [2], wherein 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).

[0026] [4]. The brain development 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).

[0027] [5]. A food product, wherein the food product contains or uses the brain development composition described in any one of [1] to [4].

[0028] [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.

[0029] [7]. The food according to [5] or [6], wherein the food may be in solid form, semi-solid form or liquid form.

[0030] [8]. Application of a composition containing three essential components—human milk oligosaccharide component, whey protein component, and phospholipid component—in the preparation of a brain development composition that promotes the development of the central nervous system and / or helps improve memory.

[0031] [9]. Application of a composition containing three essential components—human milk oligosaccharide component, whey protein component, and phospholipid component—in the preparation of foods that promote the development of the central nervous system and / or help improve memory.

[0032]

[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.

[0033]

[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.

[0034]

[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).

[0035] The effects of the invention

[0036] 1) This invention provides a brain development composition that promotes human milk oligosaccharide, comprising human milk oligosaccharide component, whey protein component and phospholipid component, which can effectively promote brain development.

[0037] 2) The composition provided by this invention can promote brain development. Specifically, the composition provided by this invention has broad application prospects in improving memory and brain development, and plays an important role in various cellular mechanisms during brain development, such as controlling neuronal maturation, synapse formation, myelination, and protecting brain cells.

[0038] 3) In in vitro models, brain cell cultures treated with the composition provided by the present invention increased the number of OPCs, promoted differentiation or maturation into OLs, and improved the myelination properties of OLs, with good synergistic effects among the components.

[0039] 4) The human milk oligosaccharide component, whey protein component, and phospholipid component in the composition of the present invention, especially the components of 3'-SL, 6'-SL, lactalbumin, and milk phospholipid, can achieve a synergistic effect when used in a limited amount, promoting neuronal maturation and synapsis, further increasing the expression of proteins related to oligodendrocyte maturation and differentiation, and promoting the expression of myelin maturation-related proteins in brain tissue; it can also significantly promote the increase of axon density, synapse-related protein expression, and neurotransmitter increase in the hippocampus and cortex.

[0040] 5) The human milk oligosaccharide component, whey protein component, and phospholipid 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.

[0041] 6) The human milk oligosaccharide component, whey protein component, and phospholipid component in the composition of the present invention, when used in a limited amount, can protect brain cells and significantly protect brain cells by reducing the content of lipid metabolites in brain tissue.

[0042] 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

[0043] Figure 1A The effect of different brain development 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 image indicate experimental groups.

[0044] Figure 1B The effect of different brain development 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 image indicate experimental groups.

[0045] Figure 2 The effect of different brain development 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.

[0046] Figure 3 The image shows the effects of different brain development composition samples on nerve cells cultured in vitro for 30 days - MBP immunostaining map (green). Scale bar: 50 μm; numbers in the figure indicate experimental groups.

[0047] Figure 4 The image shows a magnetic resonance T1-weighted image; the numbers in the image indicate the experimental group.

[0048] Figure 5 The magnetic resonance DTI image is shown; the numbers in the image indicate the experimental group.

[0049] Figure 6 The magnetic resonance (MRS) image is shown; the numbers in the image indicate the experimental group. Detailed Implementation

[0050] 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.

[0051] 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.

[0052] 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.

[0053] In this specification, the word "may" has two meanings: to perform a certain process and not to perform a certain process.

[0054] 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.

[0055] 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.

[0056] In this invention, the term "infant" is used to refer to the human group aged 0 to 6 months.

[0057] In this invention, the term "older infant" refers to the human group aged 6 to 12 months.

[0058] In this invention, the term "infant" is used to refer to the human group aged 12 to 36 months.

[0059] In this invention, the term "infant" refers to the human group under the age of 3 years.

[0060] 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.

[0061] In this manual, the term "adult" refers to a person who is 18 years of age or older.

[0062] In this manual, the term "teenager" refers to people aged 7-40.

[0063] In this manual, the term "middle-aged person" refers to a person aged 41-65.

[0064] 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.

[0065] In this specification, the terms "optional" or "optional" are used to indicate the use or omission of certain substances, components, procedures, application conditions, etc.

[0066] <First Aspect>

[0067] In a first aspect, the present invention provides a brain development composition comprising the essential components of a human milk oligosaccharide component, a whey protein component, and a phospholipid component; and wherein the mass ratio of the human milk oligosaccharide component, the whey protein component, and the phospholipid component in the composition is (0.1–15):(500–1):(1000–0.5). This brain development composition exhibits a significant synergistic effect and can promote the development of the central nervous system, such as the brain.

[0068] (Human milk oligosaccharide components)

[0069] 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.

[0070] 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.

[0071] 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.

[0072] 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.

[0073] 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.

[0074] 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.

[0075] In some embodiments, the human milk oligosaccharides of the present invention are one or more of 3'-sialylated lactose, 6'-sialylated lactose, 4'-galactosyllactose, 3'-galactosyllactose, 2'-fucosyllactose, lactose-N-tetrasaccharide, lactose-difucosyltetrasaccharide, lactose-N-neotetrasaccharide, 6'-galactosyllactose, lactose-N-fucopentose-I, and others.

[0076] In some preferred embodiments, the human milk oligosaccharide component comprises 3'-sialylated lactose (3'-SL) and 6'-sialylated lactose (6'-SL).

[0077] The human milk oligosaccharide component of the present invention can exist in a liquid, semi-solid, or solid state. When the human milk oligosaccharide component of the present invention exists in a liquid state, in some preferred embodiments, the concentration of the human milk oligosaccharide component in the brain development composition is 0-5 mg / mL.

[0078] In some preferred embodiments, the concentration of the human milk oligosaccharide component is 0-3 mg / mL.

[0079] In some more preferred embodiments, the concentration of 3'-sialylated lactose (3'-SL) in the brain development composition is 0-1 mg / mL, for example: 0.01 mg / mL; 0.05 mg / mL; 0.25 mg / mL; 0.5 mg / mL.

[0080] In some more preferred embodiments, the concentration of 6'-sialylated lactose (6'-SL) in the brain development composition is 0-2 mg / mL, for example 0.01 mg / mL; 0.02 mg / mL; 0.05 mg / mL; 0.1 mg / mL; 0.25 mg / mL; 0.5 mg / mL; 1 mg / mL.

[0081] 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).

[0082] 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, 1:5, 1:10, etc.

[0083] In some further preferred embodiments, the concentration of 3'-sialylated lactose (3'-SL) in the brain development composition is 0-0.5 mg / mL, for example: 0.05 mg / mL; 0.25 mg / mL; 0.5 mg / mL.

[0084] In some further preferred embodiments, the concentration of 6'-sialylated lactose (6'-SL) in the brain development composition is 0-1 mg / mL, for example: 0.1 mg / mL; 1 mg / mL.

[0085] 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 (1-5):(5-1). For example, the mass ratio of 3'-sialylated lactose (3'-SL) to 6'-sialylated lactose (6'-SL) is 5:2, 1:2, etc. When the 3'-sialylated lactose and 6'-sialylated lactose in the human milk oligosaccharide component are used in the above-mentioned proportions and amounts, they can have a synergistic effect. Furthermore, when used in combination with whey protein and phospholipid components, the brain development composition achieves an even better synergistic effect in promoting the central nervous system, such as brain development.

[0086] (Whey protein component)

[0087] There are no particular limitations on the whey protein components; they can be selected from commonly used whey protein powders in the field, such as whey protein concentrate, whey protein isolate, desalted whey protein powder, or any combination thereof.

[0088] In some specific implementation schemes, the whey protein powder rich in lactalbumin refers to a lactalbumin product made from raw milk and its products through processes such as defatting, acidification, membrane separation, concentration, sterilization, and drying.

[0089] Alpha-lactalbumin (abbreviated as lactalbumin) is a high-quality protein in whey protein, rich in various essential amino acids, especially cysteine ​​and tryptophan.

[0090] Whey protein can be separated from animal milk through centrifugation, membrane separation, concentration, or it can be obtained directly from commercially available products.

[0091] The whey protein component of the present invention can be present in a liquid, semi-solid, or solid state. When the whey protein component of the present invention is present in a liquid state, in some preferred embodiments, the concentration of the whey protein component in the brain development composition is 0-50 mg / mL.

[0092] In some more preferred embodiments, the concentration of the lactalbumin in the brain development composition is 0-25 mg / mL, for example: 0.1 mg / mL; 1 mg / mL; 2.5 mg / mL; 10 mg / mL; 25 mg / mL.

[0093] (Phospholipid components)

[0094] In the context of this invention, the term "phospholipid" is a collective term for all its components. Phospholipids, also known as phospholipids or phospholipid lipids, refer to lipids containing phosphoric acid and belong to complex lipids. Phospholipids are composed of five elements: C, H, O, N, and P. They are the main components of biological membranes, characterized by producing a mixture containing fatty acids and phosphoric acid upon hydrolysis. Phospholipids are amphoteric molecules, with one end being a hydrophilic nitrogen- or phosphorus-containing head and the other end being a hydrophobic (lipophilic) long hydrocarbon chain. For this reason, the hydrophilic ends of phospholipid molecules are close to each other, and the hydrophobic ends are close to each other. They often form the phospholipid bilayer, i.e., the structure of the cell membrane, together with other molecules such as proteins, glycolipids, and cholesterol.

[0095] In some embodiments, the phospholipid is a plant-derived phospholipid and / or an animal-derived phospholipid.

[0096] In some embodiments, the phospholipid may be an animal-derived phospholipid.

[0097] Furthermore, there are no particular limitations on the source of the phospholipids of the present invention in principle. They can generally be obtained by extraction from animal milk or its products. In some preferred embodiments, such animal milk or its products can be cow milk, sheep milk, camel milk, horse milk or dairy products based on them (e.g., cheese), etc. More preferably, they can be extracted from cow milk, including bovine colostrum or regular bovine milk.

[0098] In some implementations, milk phospholipids may be phospholipids derived from bovine and / or sheep milk.

[0099] In some embodiments, milk phospholipids may be provided in the following forms or may be derived from the following: protein powder containing milk phospholipids, and / or milk-derived phospholipids.

[0100] In some specific embodiments, the phospholipids described in this invention are milk phospholipids produced by Tatua of New Zealand (TATUA).

[0101] The phospholipid component of the present invention can exist in a liquid, semi-solid, or solid state. When the phospholipid component of the present invention exists in a liquid state, in some preferred embodiments, the concentration of the phospholipid component in the brain development composition is 0-20 mg / mL.

[0102] In some more preferred embodiments, the concentration of the lactalbumin in the brain development composition is 0-10 mg / mL, for example: 0.1 mg / mL; 1 mg / mL; 2.5 mg / mL; 10 mg / mL.

[0103] This invention verifies, from a (cell)biological perspective, the biological performance of human milk oligosaccharide components, whey protein components, and phospholipid components under different ratios. It finds that the combination of the three substances can promote the development of the central nervous system, such as the brain, especially promoting the proliferation of oligodendrocyte progenitor cells (OPCs), and further promoting their maturation and differentiation into oligodendrocytes (OLs). Then, from an (animal)biological perspective, it verifies that the ratio of human milk oligosaccharide components, whey protein components, and phospholipid components is a safe dosage, especially under specific ratios, which has the effect of promoting the development of the central nervous system, such as the brain, and / or helping to improve memory.

[0104] (Composition)

[0105] The brain development composition of the present invention comprises at least the human milk oligosaccharide component, whey protein component, and phospholipid component 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.

[0106] 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.

[0107] The brain development composition of the present invention can exist in a liquid state, or in a semi-solid or solid state.

[0108] When the brain development composition of the present invention is present in a liquid form, in some preferred embodiments, the concentration of the human milk oligosaccharide in the brain development composition is 0-5 mg / mL, more preferably, the concentration of the human milk oligosaccharide in the brain development composition is 0-2 mg / mL, for example: the concentration of human milk oligosaccharide is 0.03 mg / mL; 0.06 mg / mL; 0.15 mg / mL; 0.55 mg / mL; 1.5 mg / mL.

[0109] In some preferred embodiments, the concentration of the whey protein component in the brain development composition is below 50 mg / mL. More preferably, the concentration of the whey protein component in the brain development composition is not higher than 25 mg / mL, for example: the concentration of the whey protein component is 0.1 mg / mL; 1 mg / mL; 2.5 mg / mL; 10 mg / mL; 25 mg / mL.

[0110] In some preferred embodiments, the concentration of the phospholipid component in the brain development composition is less than 20 mg / mL; more preferably, the concentration of the phospholipid component in the brain development composition is less than 10 mg / mL, for example: the concentration of the phospholipid component is 0.1 mg / mL; 1 mg / mL; 2.5 mg / mL; 10 mg / mL.

[0111] When the combination of the human milk oligosaccharide component, the whey protein component, and the phospholipid component falls within the above-mentioned range, it has the effect of promoting the development of the central nervous system and / or helping to improve memory.

[0112] To obtain a better composition for promoting central nervous system development and / or aiding in memory improvement, in some embodiments, the brain development composition is wherein the mass ratio of the human milk oligosaccharide component, the whey protein component, and the phospholipid component is (0.1–15):(500–1):(1000–0.5).

[0113] In some preferred embodiments, in the brain development composition, the mass ratio of the human milk oligosaccharide component, the whey protein component, and the phospholipid component is (1-15):(300-1):(1-1000), for example: 3:2:5; 11:50:50; 3:250:1000; 3:5:5; 3:50:2; 3:2:2.

[0114] In some more preferred embodiments, in the brain development composition, the mass ratio of the human milk oligosaccharide component, the whey protein component, and the phospholipid component is (1-15):(50-1):(0.5-50), for example: 11:50:50; 3:5:5; 3:50:2; 3:2:2.

[0115] In some specific embodiments, when the combination of the human milk oligosaccharide component, the whey protein component, and the phospholipid component is within the above-mentioned ratio range, it can promote the proliferation of oligodendrocyte progenitor cells (OPCs) and further promote their maturation and differentiation into oligodendrocytes (OLs).

[0116] (Synergistic effect)

[0117] In this invention, the combined use of the human milk oligosaccharide component, whey protein component, and phospholipid component unexpectedly revealed a synergistic effect in promoting the development of the central nervous system and / or assisting in improving memory.

[0118] When the brain development composition of the present invention is present in a liquid form, in some preferred embodiments, the concentration of the human milk oligosaccharide in the brain development composition is 0-5 mg / mL, more preferably, the concentration of the human milk oligosaccharide in the brain development composition is 0-2 mg / mL, for example, the concentration of the human milk oligosaccharide is 1.5 mg / mL.

[0119] In some preferred embodiments, the concentration of the whey protein component in the brain development composition is below 50 mg / mL; more preferably, the concentration of the whey protein component in the brain development composition is not higher than 25 mg / mL, for example, the concentration of the whey protein component is 1 mg / mL or 5 mg / mL.

[0120] In some preferred embodiments, the concentration of the phospholipid component in the brain development composition is less than 20 mg / mL; more preferably, the concentration of the phospholipid component in the brain development composition is less than 10 mg / mL, for example, the concentration of the phospholipid component is 1 mg / mL.

[0121] When the combination of the human milk oligosaccharide component, the whey protein component, and the phospholipid component falls within the above-mentioned range, it has a synergistic effect in promoting the development of the central nervous system and / or in assisting to improve memory.

[0122] In order to achieve 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, the whey protein component, and the phospholipid component in the brain development composition is (0.5-2):(30-1):(0.5-2).

[0123] In some exemplary embodiments, the mass ratio of the human milk oligosaccharide component, the whey protein component, and the phospholipid component in the brain development composition is 3:50:2; 3:2:2.

[0124] When the human milk oligosaccharide component, the whey protein component, and the phospholipid component 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.

[0125] (use)

[0126] Furthermore, the above-described compositions of the present invention have been experimentally verified to promote the development of the central nervous system, including brain development, such as at least one of neuronal maturation, synapse formation, and myelination.

[0127] In some embodiments, the above-described compositions of the present invention can promote the proliferation of oligodendrocyte precursor cells, increase neurotransmitters in the brain, promote the expression of synaptic-related proteins, increase the content of myelin in the hippocampus and cortex, and promote the production of metabolites glutamylamine and acetylaspartate. Therefore, the compositions can have the effects 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.

[0128] <Second aspect>

[0129] In a second aspect of the present invention, a food product is provided, the food product comprising or using the brain development composition described in the first aspect above.

[0130] 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.

[0131] 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.

[0132] In some specific embodiments, the food can be obtained by processing the brain development composition according to the first aspect described above.

[0133] 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.

[0134] 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.

[0135] For baked goods, this can be cakes or cookies that are mainly based on butter, eggs, and baking powder.

[0136] 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.

[0137] 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.

[0138] 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.

[0139] 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.

[0140] Animal-derived ingredients, including meat products from cattle, sheep, fish, or poultry.

[0141] 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.

[0142] 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.).

[0143] 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.

[0144] 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.

[0145] 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.

[0146] 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.

[0147] This invention does not specifically limit the types of foods that can be prepared using the above-mentioned brain development composition 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 nerves and assisting in improving memory.

[0148] In some implementations, the food is a solid dairy product or a liquid dairy product; for example, milk powder, cheese, yogurt, liquid milk, etc.

[0149] 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.).

[0150] Typically, the aforementioned foods include: infant formula, maternal formula, milk powder for middle-aged and elderly people, or nutritional or dietary supplements, etc.

[0151] <Third aspect>

[0152] This invention provides the use of a composition of the three essential components—the human milk oligosaccharide component, the whey protein component, and the phospholipid component—in the preparation of a nutritional composition that promotes the development of the central nervous system and / or helps improve memory.

[0153] The present invention also provides the use of a composition containing the three essential components of the aforementioned human milk oligosaccharide component, whey protein component, and phospholipid component in the preparation of foods that promote the development of the central nervous system and / or help improve memory.

[0154] In some specific embodiments, the nutritional composition or food can promote the development of the central nervous system and / or help improve memory.

[0155] 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.

[0156] (Development of the central nervous system)

[0157] Myelination and neuronal maturation

[0158] 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.

[0159] proliferation

[0160] In some specific implementations, promoting the proliferation of oligodendrocyte precursor cells includes increasing the number of cells expressing tetrasialic acid ganglioside (GO1c).

[0161] Myelination and myelination

[0162] 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.

[0163] In some specific implementations, promoting myelination of oligodendrocyte precursor cells includes increasing the number of cells expressing myelin-associated glycoprotein (MAG).

[0164] 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.

[0165] In some specific embodiments, the compositions of the present invention significantly increased the FA values ​​of the left hippocampus and cortex, demonstrating an effect of promoting myelin formation.

[0166] neuronal maturation

[0167] In some embodiments, the compositions of the present invention can promote the further maturation and differentiation of oligodendrocyte progenitor cells (OPCs) into oligodendrocytes (OLs).

[0168] In some specific implementations, promoting the maturation of oligodendrocyte precursor cells includes increasing the number of cells expressing myelin-binding protein (MBP).

[0169] In some specific implementations, promoting neuronal maturation includes promoting the expression of pathway-related proteins in rat brain tissue, such as myelin-associated glycoprotein (MAG), myelin basic protein (MBP), and myelin proteolipid protein (PLP).

[0170] In some specific embodiments, the present invention provides evidence that the compositions of the present invention can promote an increase in myelin content in the hippocampus and cortex by detecting longitudinal relaxation rate (R1 = 1 / T1) using magnetic resonance imaging (MRI).

[0171] Synapse

[0172] 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).

[0173] In some exemplary embodiments, the present invention detects λ1, λλ, and λλ using magnetic resonance imaging (MRI) and diffusion tensor imaging (DTI) methods, wherein λ is related to the axial diffusion coefficient and the average diffusion coefficient, and λ1 and λλ reflect the lateral diffusion coefficient.

[0174] In some specific embodiments, the compositions of the present invention significantly increase the λ1 value of the hippocampus and cortex, thus promoting an increase in axon density.

[0175] In some specific embodiments, the compositions of the present invention can promote an increase in axon density in the hippocampus and cortex.

[0176] In some embodiments, the compositions of the present invention can promote an increase in neurotransmitters in the brain.

[0177] In some embodiments, the compositions of the present invention can promote an increase in the neurotransmitter 5-hydroxytryptamine (5-HT) in the brain.

[0178] (Aids in improving memory)

[0179] In some specific implementations, the aid to improve memory includes promoting the levels of the metabolites glutamine and acetylaspartate in brain tissue. The production of glutamine and acetylaspartate can promote the formation of neurite buds and synapses, thereby playing an aid to improve memory.

[0180] In some specific embodiments, the composition can promote the reduction of metabolite lipids (Lip) in brain tissue, thus protecting brain cells.

[0181] In some implementation schemes, the specific selection and ratio of the human milk oligosaccharide component, whey protein component, and phospholipid component are as described in the preceding <First Aspect>.

[0182] Example

[0183] 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.

[0184] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention. Unless otherwise specified, the reagents, methods, and equipment used in this invention are conventional in the art.

[0185] This invention utilizes an in vitro model of primary cell culture containing neurons and OPCs to evaluate the effects of a nutrient mixture on myelination and neuronal development. A mixed nutrient solution is added to the mixed cell culture to promote the proliferation, maturation, and differentiation of OPCs into mature neurons (OLs) and / or the myelination properties of OLs. This invention first assesses the density (DIV) of OPCs after 12 days of in vitro culture. Then, at 18 and 30 days, this invention assesses OPC differentiation into OLs and OL maturation by quantifying the proportions of MAG-positive and MBP-positive cells, respectively. Finally, this invention assesses the level of myelination after 18 and 30 DIVs by evaluating the proportion of axons surrounded by the OL process in the cell culture. This method systematically and intuitively evaluates the effects of compositions of 3'-SL and 6'-SL, lactaloxides, etc., on early brain development, and has profound implications for guiding the application of these compositions.

[0186] Example 1: Cellular Experiment on the Brain-Boosting Effect of the Composition

[0187] I. Materials and Methods

[0188] (I) Instruments, Reagents and Consumables

[0189] Cell culture incubator; laser confocal fluorescence microscope; centrifuge; electronic balance; vortex mixer; water bath; tissue homogenizer.

[0190] Nerve cell culture medium; antibodies such as A2B5, MAG, and MBP.

[0191] Reagent: 6'-SL: DSM, GLYCARE 6SL9001, (Catalog No.: 5016570);

[0192] 3'-SL: DSM, GLYCARE6SL9001, (Product No.:5016567);

[0193] Olesoxime: Selleck;

[0194] Lactalbumin: Arla (Production batch number: P420215);

[0195] Milk phospholipids: TATUA (Production batch number 156152).

[0196] (II) Experimental Methods

[0197] 1. Obtaining primary neural cells

[0198] All experiments were ethically approved, and primary co-culture of neurons and neurons (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 min using trypsin (Trypsin EDTA 1X, 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 min.

[0199] 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.

[0200] 2. Nerve cell culture

[0201] 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 (brain development composition containing 3'-SL, 6'-SL, lactalbumin, and / or lactophospholipids prepared above) were added to the fresh medium.

[0202] 3. Immunohistochemical experiment

[0203] After 12, 18, and 30 DIV, cells were fixed by incubation for 5 min with a cold mixture of 95% (v / v) ethanol and 5% (v / v) acetic acid. Then, nonspecific sites were blocked for 15 min at room temperature with phosphate-buffered saline (PBS) containing 0.1% (w / w) saponin (Sigma) and 1% (v / v) FCS (GIBCO).

[0204] 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.

[0205] 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).

[0206] 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).

[0207] 4. Microscopic photography

[0208] 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.

[0209] 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.

[0210] 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.

[0211] 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).

[0212] (III) Grouping of Cell Experiments

[0213] 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 milk phospholipids, lactalbumin, and 3'-SL and 6'-SL alone or in combination on OPC population, OL maturation and differentiation, myelin formation, and neurite growth.

[0214] (IV) Statistical Analysis

[0215] Results are expressed as mean ± standard error. Data analysis software was used to perform T-tests and one-way ANOVA tests. A p-value < 0.05 was considered statistically significant.

[0216] II. Experimental Results

[0217] 1. Effects of each sample on nerve cells

[0218] 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.

[0219] Figure 1A The effects of different brain development composition samples on neural 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.

[0220] Table 1 shows the effects of 3'-SL and 6'-SL brain development composition samples on the number of A2B5 positive cells.

[0221]

[0222] 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 6 and 7 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.

[0223] In terms of components and dosages, cell experiment group 6 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 6 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 7.

[0224] Subsequently, the inventors further added lactalbumin and lactophospholipids to the composition to culture nerve cells for the next stage of experimentation. Figure 1B The effects of different brain development composition samples on nerve cells cultured in vitro for 12 days are shown in the A2B5 immunostaining map (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.

[0225] Table 2 shows the effects of different brain development composition samples containing 3'-SL, 6'-SL and / or lactalbumin and lactophospholipid on the number of A2B5 positive cells.

[0226]

[0227] The sample processing results (Table 2) showed that, compared with the blank control group, the combined effects of the two types of human milk oligosaccharides, lactalbumin, and milk phospholipids increased the number of A2B5 positive cells in each group. The results of each group were greater than those of the combination of the two human milk oligosaccharides.

[0228] 2. Effects of each sample on myelination of nerve cells

[0229] 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.

[0230] Figure 2The effects of different brain development composition samples on neural cells cultured in vitro for 18 days are shown in the MAG immunostaining map (green). Scale bar: 50 μm. The measured number of MAG-positive cells per image and the average number of MAG-positive cells per well are shown in Table 3.

[0231] Table 3 shows the effects of different brain development composition samples containing 3'-SL, 6'-SL and / or lactalbumin and lactophospholipid on the number of MAG-positive cells.

[0232]

[0233] 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.

[0234] The sample processing results showed that, compared with the blank control group, when the four components of two types of human milk oligosaccharides, lactalbumin and milk phospholipids worked together, each group could increase the number of myelinated oligodendrocytes.

[0235] 3. Effects of each sample on OPC cell maturation

[0236] 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.

[0237] Figure 3 The effect of different brain development composition samples on nerve cells cultured in vitro for 30 days is shown in the MBP immunostaining map (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.

[0238] Table 4 shows the effects of different brain development composition samples containing 3'-SL, 6'-SL and / or lactalbumin and lactophospholipids on the number of MBP-positive cells.

[0239]

[0240]

[0241] 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.

[0242] The sample processing results showed that, compared with the blank control group, when the four components of two types of human milk oligosaccharides, lactalbumin and milk phospholipids worked together, each group could increase the number of mature oligodendrocyte precursor cells.

[0243] Example 2: Animal experiments on the brain-boosting effects of the composition

[0244] I. Materials and Methods

[0245] (I) Instruments, Reagents and Consumables

[0246] 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.

[0247] Reagents: 5-HT and 5-HIAA ELISA kit (Elabscience Biotechnology Co., Ltd. (China)); 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).

[0248] (II) Experimental Methods

[0249] 1. Grouping and intervention of experimental animals

[0250] Three-week-old male SD rats (n=77) were acclimatized to the diet for one week and then randomly divided into 7 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. The food intake and body weight of the rats 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. The rat blood was centrifuged at 4℃, 3000 rpm for 15 min to obtain serum, which was then stored at -80℃.

[0251] Table 5. Test Dosage and Group

[0252]

[0253]

[0254] Note: "—" indicates that no addition was made or that there is no proportional relationship.

[0255] 2. Body weight measurement

[0256] Body weight and weekly food intake of SD rats were measured on days 0, 7, 14, 21, and 28.

[0257] 3. The ratio of animal organs and body fat content

[0258] 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.

[0259] 4. Measurement of serum and brain tissue chemical indicators

[0260] Serum samples stored at -80℃ were thawed at 4℃, and the levels of 5HT and 5HIAA in the serum were determined according to the kit instructions.

[0261] 100 mg of brain tissue sample stored at -80°C was quickly weighed and homogenized in an ice-water bath.

[0262] Centrifuge at 4℃ and 10000 rpm for 10 min and collect the supernatant. Determine the content of 5HT and 5HIAA in the homogenate supernatant.

[0263] 5. WES method for detecting synapse-related proteins and myelin-related proteins in brain tissue

[0264] 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.

[0265] 6. Magnetic resonance imaging of rat brain tissue

[0266] (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.

[0267] 7. Statistical Analysis

[0268] Results are expressed as mean ± standard error (mean ± SEM). T-test and one-way ANOVA were performed using data analysis software. A p < 0.05 was considered statistically significant, and a p < 0.01 was considered highly statistically significant.

[0269] (III) Experimental Results

[0270] 1. Effects of the composition on rat body weight, food intake, organ index, and body fat percentage

[0271] The effects of the composition on rat body weight, food intake, organ index and body fat percentage are shown in Table 6-7.

[0272] 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 rats in the control group was 324.63±12.81g; the body weight of rats in the six groups was slightly higher than that of the control group, but the difference was not significant.

[0273] Table 6. Effects of the composition on food intake in rats

[0274]

[0275] Table 6 shows that the food intake of rats in each group initially increased and then decreased throughout the experiment. Food intake was higher in the first two weeks and gradually decreased in the latter 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.

[0276] Table 7 Effects of the composition on organ index and body fat percentage in rats

[0277]

[0278] The organ indices (heart, liver, spleen, and kidney) in each group in Table 7 showed no significant differences 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 percentages in each intervention group remained unchanged, with only groups 6 and 7 showing slightly higher body fat percentages, but these differences were not statistically significant. This indicates that the selected composition did not significantly affect the body fat percentages of normal rats.

[0279] 2. Effects of the composition on the levels of 5HT and 5HIAA in rat serum and brain tissue

[0280] The levels of 5HT and 5HIAA in the serum and brain tissue of rats in each group are shown in Table 8.

[0281] Table 8. Effects of the composition on 5HT and 5HIAA levels in rat serum and brain tissue.

[0282]

[0283] Note: "**" indicates a comparison with the blank control group, p < 0.01.

[0284] 5-Hydroxytryptamine (5-HT) is an important intracellular messenger and inhibitory neurotransmitter found in the human brain and digestive tract. It is first converted from tryptophan to 5-HTP by tryptophan hydroxylase, then to 5-HT by 5-hydroxytryptamine decarboxylase, and finally to 5-hydroxyindoleacetic acid (5-HIAA) by monoamine oxidase and aldehyde dehydrogenase, and then excreted in urine.

[0285] As shown in Table 8, compared with the control group, the composition of the present invention can increase the levels of 5-HT and 5-HIAA in serum. In brain tissue, compared with the control group, the 5-HT content in groups 6 and 7 was significantly increased, and the 5-HIAA content in group 7 was significantly increased. This indicates that the combination containing α-lactalbumin can significantly increase the 5-HT content in rat brain tissue. The combination containing a medium dose of α-lactalbumin can significantly increase the 5-HIAA content in brain tissue.

[0286] Because 5-HT cannot cross the blood-brain barrier, the central and peripheral 5-HT systems are functionally separate. Studies have shown that changes in 5-HT levels are closely related to mental and neurological diseases. 5-HIAA, the final metabolite of 5-HT, can indirectly reflect changes in 5-HT levels. During neuronal development, 5-HT contributes to synapse formation, connection, and network construction, and plays a regulatory role in post-mitotic neuronal proliferation, migration, differentiation, maturation, and apoptosis. 5-HT also regulates cell adhesion molecules, injecting neuronal plasticity into the developing and adult brain. The 5-HT signaling pathway also significantly regulates adult hippocampal neurogenesis. Furthermore, 5-HT plays a role in regulating various other ligands, including hormones and neurotransmitters such as dopamine, adrenaline and γ-aminobutyric acid (GABA), cortisol, prolactin, acetylcholine, and oxytocin. In summary, 5-HT, as a crucial neurotransmitter, regulates a wide range of functions, including mood, cognition, anxiety, learning, memory, reward processing, and sleep.

[0287] 3. Expression of pathway-related proteins in the brain tissue of rats in each group

[0288] The expression of pathway-related proteins in the brain tissue of rats in each group is shown in Tables 9 and 9-1.

[0289] Table 9. Effects of the composition on the expression of pathway-related proteins in rat brain tissue.

[0290]

[0291]

[0292] Note: "*" indicates that compared with the blank control group, p < 0.05; "**" indicates that compared with the blank control group, p < 0.01.

[0293] Table 9-1 Effects of the composition on the expression of pathway-related proteins in rat brain tissue

[0294]

[0295] Note: "**" indicates a comparison with the blank control group, p < 0.01.

[0296] 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.

[0297] 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.

[0298] 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.

[0299] 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).

[0300] The sample processing results showed that, compared with the blank control group, the medium-dose groups of lactalbumin, when used alone within the tested dose range, significantly increased the expression of synaptic-related proteins (SYP, PSD, GAP-43, and BDNF). The 3'-SL+6'-SL combination and lactalcohol themselves did not promote the expression of synaptic-related proteins, but when used in combination with lactalbumin (groups 6-7), they significantly increased the promoting effect of lactalbumin on the expression of synaptic-related proteins. This indicates that the 3'-SL+6'-SL combination and lactalcohol have a synergistic effect with lactalbumin and can synergistically promote the expression of synaptic-related proteins.

[0301] 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, an important protein in myelination, is responsible for intracellular myelin compaction, which is achieved by binding the cytoplasmic surface of the myelin membrane together.

[0302] The combined use of 3'-SL+6'-SL, milk phospholipids, and lactalbumin (groups 6-7) significantly increased the expression of myelin-related proteins (Table 9-1). Furthermore, the 3'-SL+6'-SL combination and milk phospholipids synergistically promoted MBP protein expression. This indicates that the use of the 3'-SL+6'-SL, milk phospholipids, and lactalbumin combination helps to synergistically promote myelin compaction in animals.

[0303] 4. Effects of the composition on magnetic resonance imaging results of rat brain tissue

[0304] Table 10 Effects of the composition on T1 in rat brain tissue

[0305]

[0306]

[0307] Note: "**" indicates a comparison with the blank control group, p < 0.01.

[0308] Table 10-1 Effects of the composition on T1 in left rat cortical tissue

[0309] Animal experimental group Dosage group Left cortex (ms) 1 Blank control group 2045.71±40.46 2 3'-SL+6'-SL <![CDATA[2139.42±33.57 * ]]> 3 Low dose of lactalbumin <![CDATA[2145.17±57.08 * ]]> 5 Milk phospholipids <![CDATA[2201.79±64.43 ** ]]> 6 3'-SL + 6'-SL + milk phospholipids + low-dose lactalbumin 2030.64±99.58

[0310] Note: "*" indicates that compared with the blank control group, p < 0.05; "**" indicates that compared with the blank control group, p < 0.01.

[0311] 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 in the central nervous system as electrical signals. The myelin sheath consists of repetitive units of a tight oligodendrocyte membrane, a double-layered tissue separated by water and a 3-4 nm extracellular space. Unlike membranes in other cell types, it is typically characterized by a lipid and protein content of 50% by weight; the myelin sheath membrane is composed 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) (where R1 = 1 / T1, indicating a shortened relaxation time) due to the interaction of free water molecules with macromolecules. Therefore, T1 localization is a sensitive quantitative method for myelin formation and has been studied in preterm infants, normal infants, and children and adolescents.

[0312] Table 10 shows that in the right hippocampus, compared with the blank control group, the low- and medium-dose lactalbumin groups significantly increased myelin content when used alone within the tested dose range. The combination of 3'-SL+6'-SL, lactalbumin, and lactalbumin (groups 6-7) all significantly increased myelin content. Specifically, in group 6, 3'-SL+6'-SL and lactalbumin alone did not show significant effects, but the combination with low-dose lactalbumin significantly enhanced the effect compared to low-dose lactalbumin alone, indicating a synergistic effect in group 6.

[0313] As shown in Table 10-1, in the left cortex, compared with the blank control group, in group 6, the low-dose groups of 3'-SL+6'-SL, lactalcohol, and lactalbumin did not show any effect when used alone within the tested dose range. The combined use of 3'-SL+6'-SL, lactalcohol, and lactalbumin (group 6) increased the myelin content. The effect was enhanced compared to the individual dose groups, indicating that the combination in group 6 has a synergistic effect.

[0314] 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 and average diffusion coefficients, while λ2 and λ3 reflect the lateral diffusion coefficients.

[0315] Table 11 Effects of the composition on DTI in the right hippocampus of rats

[0316]

[0317] Table 12 Effects of the composition on DTI in the left hippocampus of rats

[0318]

[0319] Note: "**" indicates a comparison with the blank control group, p < 0.01.

[0320] Table 13 Effects of the composition on DTI in the right cortex of rats

[0321]

[0322]

[0323] Note: "*" indicates that compared with the blank control group, p < 0.05; "**" indicates that compared with the blank control group, p < 0.01.

[0324] Table 14 Effects of the composition on left cortical DTI in rats

[0325]

[0326] Note: "*" indicates that compared with the blank control group, p < 0.05; "**" indicates that compared with the blank control group, p < 0.01.

[0327] As shown in Tables 11 to 14, in the left hippocampus, left cortex, and right cortex, group 6 (3'-SL + 6'-SL + lactophospholipid dose + low-dose lactalbumin) significantly increased FA and λ1 values, demonstrating a significant effect in promoting myelin formation and increasing axonal density. Figure 6 ).

[0328] Table 15 Effects of the composition on MRS in rat brain tissue

[0329]

[0330] Note: "*" indicates that compared with the blank control group, p < 0.05; "**" indicates that compared with the blank control group, p < 0.01.

[0331] 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.

[0332] As shown in Table 15, the composition has no effect on the content of glutamine, inositol, taurine, choline, and lactic acid. Therefore, the composition will not affect the normal metabolism of brain tissue.

[0333] Acetyl aspartate (NAA): It is involved in protein and fat synthesis, maintaining intracellular cation concentration, and the passage of cations such as potassium, sodium, and calcium through cells, as well as maintaining the excitability of nerve membranes. It exists only within neurons and not in glial cells, serving as a marker of neuronal density and survival. The amount present reflects the functional status of neurons, and the degree of reduction reflects the extent of damage. Table 15 shows that group 7 significantly increased NAA content compared to the control group, suggesting that group 7 (3'-SL + 6'-SL + lactophospholipid dose + medium dose of lactalbumin) may increase neuronal density.

[0334] The lipid (Lip) peak was most commonly seen in necrotic brain cells. As shown in Table 15, compared with the control group, group 6 had a significantly reduced lipid content, suggesting that group 6 has a protective effect on brain cells.

[0335] In summary, this invention provides insights into the application of two human milk oligosaccharides, lactalbumin, and milk phospholipids in brain development, particularly in neural development, offering new avenues for the development of future functional foods. These two human milk oligosaccharides, lactalbumin, and milk phospholipids show great promise in improving memory and brain development. Studies have found that the extracellular environment plays a crucial role in regulating brain homeostasis and controlling myelination during central nervous system development. Deficiencies in key nutrients can significantly impact brain development. Our research demonstrates that, in in vitro models, brain cell cultures treated with a combination of these two human milk oligosaccharides, lactalbumin, and milk phospholipids 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.

[0336] 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.

[0337] 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 brain development composition comprising a human milk oligosaccharide, characterized in that, The brain development composition comprises essential components as follows: a human milk oligosaccharide component, a whey protein component, and a phospholipid component; and, in the brain development composition, the mass ratio of the human milk oligosaccharide component, the whey protein component, and the phospholipid component is (0.1-15):(500-1):(1000-0.5).

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

4. The brain development 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) and 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 brain development composition according to any one of claims 1-4.

6. The food product of claim 5, wherein, The food comprises 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 essential components of a human milk oligosaccharide component, a whey protein component, and a phospholipid component 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 essential components of a human milk oligosaccharide component, a whey protein component, and a phospholipid component in the preparation of a food for promoting central nervous system development and / or assisting in improving memory.

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

11. Use according to claim 10, characterized in that, The central nervous system development comprises 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 comprises promoting the expression of synapse-related proteins, preferably, the synapse-related proteins comprise 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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