Composition for synergistically promoting brain development

Abstract

The invention provides a composition for synergistically promoting brain development. The invention provides a composition. The composition is composed of 2 '-FL and 3'-SL in a mass ratio of (2.5: 1)-(15: 1). The invention further provides food and medicine containing the composition, application of the composition in preparation of food or medicine for promoting brain development, application of the food or medicine for promoting brain development and a method for promoting brain development.

Description

Technical Field

[0001] The present invention belongs to the field of brain development. More specifically, the present invention relates to a composition that synergistically promotes brain development. Background Art

[0002] Brain development, especially the development of the nervous system, is crucial for the growth of infants, children, and adolescents. Imperfect brain development may lead to various symptoms such as inattention, hyperactivity disorder, anxiety disorder, and learning and memory difficulties.

[0003] Early studies have shown that ginkgo biloba extract (GBE) contains 24% flavonoids and 6% terpene lactones, and can enhance long-term potentiation by regulating the BDNF-TrkB signaling pathway in the hippocampus; cistanche polysaccharide promotes the differentiation of oligodendrocytes by activating the mTOR pathway. The ginkgo biloba and cistanche tablets developed based on the composition of ginkgo biloba extract and cistanche polysaccharide have the effects of enhancing memory, supplementing brain nutrition, and enhancing the vitality of brain cells, and can promote brain development. However, it has been found that the promoting effect of ginkgo biloba and cistanche tablets on brain development is still limited, and the effect on healthy people is not significant; ginkgolic acid (an allergen) may remain during the extraction of ginkgo biloba, posing an allergic risk; the growth cycle of ginkgo trees is long (leaves can be used after more than 10 years), and the limited high-quality raw materials lead to a high price. These problems limit the application of ginkgo biloba extract.

[0004] Therefore, there is an urgent need in this field to explore a composition that can significantly promote brain development. Summary of the Invention

[0005] The purpose of the present invention is to provide a composition that synergistically promotes brain development.

[0006] In the first aspect of the present invention, a composition is provided, which is composed of 2′-FL and 3′-SL with a mass ratio of (2.5:1) to (15:1).

[0007] In one or more embodiments, the amount of 2′-FL is 2.5 to 14 parts by mass, preferably 3 to 13 parts by mass, 4 to 12 parts by mass, 5 to 11 parts by mass, 6 to 10 parts by mass, 7 to 9 parts by mass, and more preferably 7 to 8 parts by mass, relative to 1 part by mass of 3′-SL.

[0008] In the second aspect of the present invention, a food is provided, which contains the composition according to any embodiment of the present invention.

[0009] In one or more embodiments, the food includes food products, food semi-finished products, food additives, and food supplements.

[0010] In one or more embodiments, the food further comprises a food pharmaceutically acceptable material.

[0011] In one or more embodiments, the food pharmaceutically acceptable material comprises: nutritional additives, ingredients both used as food and medicine, excipients and / or adjuvants.

[0012] In one or more embodiments, the nutritional additives include one or more of dietary fiber, prebiotics, proteins, lipids, minerals, and vitamins.

[0013] In one or more embodiments, the ingredients both used as food and medicine include one or more of red dates, hawthorns, wolfberries, longans, lilies, poria cocos, and dried tangerine peels.

[0014] In one or more embodiments, the excipients or adjuvants include one or more of calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.

[0015] In one or more embodiments, the food is infant food, children's food, juvenile food, adolescent food, young adult food, adult food, middle-aged food, or elderly food.

[0016] In one or more embodiments, the infants include babies, older babies, and toddlers.

[0017] In one or more embodiments, the food is infant formula milk powder, baby complementary food, children's formula milk powder, children's snacks, pregnant women's formula milk powder, middle-aged and elderly milk powder, or nutritional or dietary supplements.

[0018] In a third aspect of the present invention, there is provided a drug which comprises an effective amount of the composition according to any one of the embodiments of the present invention, and a pharmaceutically acceptable carrier.

[0019] In one or more embodiments, in the composition: the effective amount of 2′-FL is 500 - 1000 μg / mL, and / or the effective amount of 3′-SL is 45 - 2000 μg / mL.

[0020] In a fourth aspect of the present invention, there is provided the use of the composition according to any one of the embodiments of the present invention in the preparation of a drug for promoting brain development.

[0021] In a fifth aspect of the present invention, there is provided the use of the drug according to any one of the embodiments of the present invention in promoting brain development, or the use of the food according to any one of the embodiments of the present invention in promoting brain development for non-therapeutic purposes.

[0022] In one or more embodiments, the promoting brain development is to improve brain, nerve, and / or cognitive development.

[0023] In one or more embodiments, the promoting of brain development includes: preventing or improving brain developmental retardation, nervous system developmental retardation, and / or cognitive developmental retardation.

[0024] In one or more embodiments, the promoting of brain development includes enhancing cognitive maturity and promoting the expression of genes related to brain development.

[0025] In one or more embodiments, the genes related to brain development include bdnf and gdnfa.

[0026] A sixth aspect of the present invention provides a method for promoting brain development for non-therapeutic purposes, the method comprising: administering the composition according to any one of the embodiments of the present invention to an individual in need, or administering the food according to any one of the embodiments of the present invention or the drug according to any one of the embodiments of the present invention to an individual in need.

[0027] In one or more embodiments, the promoting of brain development is as defined in any one of the embodiments of the present invention.

[0028] Other aspects of the present invention will be apparent to those skilled in the art from the present disclosure. Brief Description of the Drawings

[0029] Figure 1A Typical graphs of zebrafish movement trajectories after treatment with 2′-FL and 3′-SL, where the blue box is the blue area of the plus maze, which is the area for quantification.

[0030] Figure 1B Percentage of movement in the blue area of zebrafish after treatment with 2′-FL and 3′-SL, compared with the normal control group, *p < 0.05, **p < 0.01.

[0031] Figure 2A Typical graphs of zebrafish movement trajectories after treatment with a composition of 2′-FL and 3′-SL, where the blue box is the blue area of the plus maze, which is the area for quantification.

[0032] Figure 2B Percentage of movement in the blue area of zebrafish after treatment with a composition of 2′-FL and 3′-SL, compared with the normal control group, *p < 0.05, **p < 0.01, ***p < 0.001; compared with Formula 1, # p < 0.05, ## p < 0.01; compared with Formula 2, && p < 0.01; compared with Formula 3, @ p < 0.05, @@ p < 0.01; compared with Formula 4,$ p < 0.05.

[0033] Figure 3 After treatment with the composition of 2′-FL and 3′-SL, the relative expression level of the zebrafish bdnf gene, compared with the normal control group, **p < 0.01, ***p < 0.001; compared with Formula 1, # p < 0.05, ### p < 0.001; compared with Formula 2, & p < 0.05, && p < 0.01, &&& p < 0.001; compared with Formula 3, @ p < 0.05, @@ p < 0.01, @@@ p < 0.001; compared with Formula 4, $ p < 0.05, $$$ p < 0.001.

[0034] Figure 4 After treatment with the composition of 2′-FL and 3′-SL, the relative expression level of the zebrafish gdnfa gene, compared with the normal control group, **p < 0.01, ***p < 0.001; compared with Formula 1, # p < 0.05, ## p < 0.01; compared with Formula 2, && p < 0.01, &&& p < 0.001; compared with Formula 3, @ p < 0.05, @@ p < 0.01, @@@ p < 0.001; compared with Formula 4, $ p < 0.05, $$ p < 0.01.

[0035] Figure 5A After treatment with the composition of 2′-FL, 3′-SL and other compositions, the typical diagram of the zebrafish movement trajectory, where the blue square is the blue area of the plus maze, which is the quantitative area.

[0036] Figure 5B After treatment with the composition of 2′-FL, 3′-SL and other compositions, the proportion of movement in the blue area of the zebrafish, compared with the normal control group, ***p < 0.001; compared with the group of 3 raw material ratios, # p < 0.05. Detailed implementation methods

[0037] Unless otherwise specified, the meanings of the terms in this application are the same as those commonly understood by those skilled in the art. For the various experimental techniques mentioned herein, those skilled in the art can refer to various textbooks, literature, and commercial product descriptions, or they can also refer to the specific examples given in the embodiment section of this application.

[0038] In the present invention, unless otherwise specified, all ratios are by weight. It should be understood that the specific numerical values (such as ratios) given herein should not only be understood as individual numerical values, but should also be considered as providing the end point values of a certain range, and can be combined with each other to provide other ranges. For example, when the weight ratio of 2′-FL and 3′-SL is disclosed as (2.5:1)~(15:1), it is equivalent to disclosing that the weight ratio of the two can be 2.5:1, or 15:1, or any ratio within this range, such as 7:1.

[0039] Some exemplary embodiments of this application are described in detail below. It should be understood that these detailed descriptions are only for those skilled in the art to more clearly understand the content of this application, and are not intended to limit in any way. Those skilled in the art can make various changes and variations to the described embodiments.

[0040] Through in-depth research, the present inventors found that only the composition of 2′-FL and 3′-SL has a synergistic effect on promoting brain development when within a specific ratio range of (2.5:1)~(15:1), while the composition with 3-FL added on the basis of 2′-FL and 3′-SL, and the composition outside this ratio range, cannot play a good role in promoting brain development. Therefore, the present invention provides a composition composed of 2′-FL and 3′-SL, and also provides its application in promoting brain development.

[0041] Composition

[0042] The present invention provides a composition, which is composed of 2′-FL and 3′-SL with a mass ratio of (2.5:1)~(15:1).

[0043] As used in the present invention, "2′-FL" refers to 2'-fucosyllactose, which is a trisaccharide compound formed by a lactose core structure in which β-D-galactose is linked to D-glucose through a 1-4 glycosidic bond and fucose is linked to the C2' hydroxyl site through an α1-2 glycosidic bond, and its chemical formula is C 18 H 32 O 15 , with a molecular weight of 488.44 g / mol and a CAS number of 41263-94-9.

[0044] As used in the present invention, "3'-SL" refers to 3'-sialyllactose, which is a tetrasaccharide compound formed by connecting N-acetylneuraminic acid (Neu5Ac) to the lactose core structure formed by connecting β-D-galactose and D-glucose through a 1-4 glycosidic bond at the C3' hydroxyl site, and its chemical formula is C 23 H 39 NO 19 , with a molecular weight of 633.56 g / mol and a CAS number of 35890-38-1.

[0045] The present invention discovers that only when the composition of 2'-FL and 3'-SL is within a specific weight ratio range of (2.5:1) to (15:1), it has a synergistic effect on promoting brain development. Therefore, the mass ratio of 2'-FL to 3'-SL can be (2.5:1) to (15:1), such as (2.5:1) to (13:1), (2.5:1) to (12:1), (2.5:1) to (10:1), (2.5:1) to (9:1), (2.5:1) to (8:1), (2.5:1) to (7:1), (5:1) to (15:1), (5:1) to (10:1), or (5:1) to (7:1). It can also be said that when 2'-FL and 3'-SL are used in combination, relative to 1 part by mass of 3'-SL, the amount of 2'-FL can be 2.5 to 15 parts by mass, such as 2.5 to 14 parts by mass, 3 to 13 parts by mass, 4 to 12 parts by mass, 5 to 11 parts by mass, 6 to 10 parts by mass, 7 to 9 parts by mass, 7 to 8 parts by mass, or a range defined by any two point values. When the weight ratio of 2'-FL to 3'-SL is within the above range, its effect on promoting brain development is more significant, and there is a synergistic effect between the two.

[0046] In some embodiments, in the composition, the concentration of 2'-FL can be 500 to 1000 μg / mL, such as 500 to 900 μg / mL, 500 to 800 μg / mL, 500 to 750 μg / mL, 500 to 720 μg / mL, 500 to 710 μg / mL, 510 to 710 μg / mL, 520 to 710 μg / mL, or 530 to 710 μg / mL. In other embodiments, in the composition, the concentration of 3'-SL can be 45 to 2000 μg / mL, such as 45 to 1000 μg / mL, 45 to 800 μg / mL, 45 to 600 μg / mL, 45 to 500 μg / mL, 45 to 300 μg / mL, 45 to 250 μg / mL, or 45 to 220 μg / mL.

[0047] The present invention also provides a method for preparing the composition, the method comprising the step of mixing 2′-FL and 3′-SL.

[0048] Food

[0049] The present invention also provides a food comprising the composition.

[0050] The term "food" means an article or substance that can be ingested by an individual into their body, and includes finished food products, semi-finished food products, food additives, food supplements, and health products.

[0051] In the present invention, the term "individual" refers to any animal of interest. In some embodiments, the individual is a mammal, such as humans, monkeys, cows, sheep, horses, pigs, goats, dogs, cats, mice, rats.

[0052] The composition of the present invention can be formulated into a food using standard techniques well known to those of ordinary skill in the art. For example, the composition can be directly added to a food pharmaceutically acceptable material, or it can be used to prepare an intermediate composition (e.g., a food additive or premix) suitable for subsequent addition to a food pharmaceutically acceptable material.

[0053] The term "food pharmaceutically acceptable material" refers to nutritional additives (e.g., dietary fiber, prebiotics, proteins, lipids, minerals, vitamins) that can be applied to foods, ingredients homologous in medicine and food (e.g., red dates, hawthorn, wolfberries), excipients or adjuvants (e.g., calcium carbonate, calcium phosphate, various sugars and starches, cellulose derivatives, gelatin, vegetable oils, polyethylene glycol), etc.

[0054] The food of the present invention can be in any form suitable for oral administration, such as powder, tablet, capsule, granule, solution, suspension, etc.

[0055] The food of the present invention is generally applicable to people of any age group, such as infants and young children (including infants, older infants, and toddlers), children, juveniles, adolescents, young adults, adults, middle-aged people, or the elderly. The term "infant" refers to a person aged 0 to 6 months. The term "older infant" refers to a person aged 6 to 12 months. The term "toddler" refers to a person aged 12 to 36 months. The term "infants and young children" refers to a person aged 0 - 36 months. The term "child" refers to a person aged 3 - 6 years. The term "juvenile" refers to a person aged 7 - 17 years. The term "adult" refers to a person aged 18 years or older. The term "young adult" refers to a person aged 18 - 40 years. The term "adolescent" refers to a person aged 7 - 40 years. The term "middle-aged person" refers to a person aged 41 - 65 years. The term "elderly person" or "the elderly" refers to a person aged 65 years or older.

[0056] In some embodiments, the food may be food for infants and young children (such as infant food, older infant food, toddler food), children's food, adolescent food, or adult food, such as infant formula (such as infant formula milk powder, toddler formula milk powder), baby complementary food, nutritional or dietary supplements, children's formula milk powder, children's snacks, pregnant women's formula milk powder, or middle-aged and elderly milk powder.

[0057] In the food, the addition amount of the composition may be such that the food can promote brain development. In one embodiment, relative to the total mass of the food, the mass percentage of the composition may be 0.001 - 80%, preferably 0.01 - 50%, for example, it may be 0.1 - 30%, 1 - 20%. In addition to 2′-FL and 3′-SL, the food may also contain other ingredients, such as other proteins / amino acids, carbohydrates, fats, vitamins, minerals, and other food - acceptable materials.

[0058] Drug

[0059] The present invention also provides a drug comprising the composition. The drug may further include a pharmaceutically acceptable carrier.

[0060] As used herein, "pharmaceutically acceptable carrier" refers to those carriers that do not cause significant irritation to an organism and do not impair the biological activity and properties of the reagents in the administered pharmaceutical composition, such as but not limited to: solvents, buffers, emulsifiers, suspending agents, decomposers, disintegrating agents, dispersing agents, binding agents, excipients, stabilizing agents, chelating agents, diluents, gelling agents, preservatives, wetting agents, lubricants, absorption delaying agents or liposomes. In some embodiments, a pharmaceutically acceptable carrier may be an inert substance added to a pharmaceutical composition to further facilitate the administration of the reagent, such as calcium carbonate, calcium phosphate, various sugars and starches, cellulose derivatives, gelatin, vegetable oils, polyethylene glycol, etc.

[0061] The drugs of the present invention can be formulated into any suitable dosage form for administration by oral or other means. Dosage forms suitable for oral administration include but are not limited to: sterile powders, tablets, troches, lozenges, pellets, capsules, dispersible powders or granules, solutions, suspensions, drops, emulsions, syrups, elixirs or slurries. The drugs of the present invention can also be stored in a sterilized device suitable for injection or infusion.

[0062] The compositions of 2′-FL and 3′-SL in the drug are usually present in an effective amount (such as a therapeutically effective amount, a prophylactically effective amount). An effective amount is a dosage sufficient to improve or in some way alleviate the symptoms associated with a disease, such as a dosage that effectively improves or eliminates one or more disease conditions, and can be determined according to the age, gender, physical condition, etc. of the subject. The dosage may cure the disease, but administration is usually for improving the symptoms of the disease. Repeated administration is generally required to achieve the desired symptom improvement.

[0063] In a specific embodiment of the present invention, zebrafish is used as an experimental animal, and a dosing regimen for promoting brain development with a composition of 2′-FL and 3′-SL is proposed. When zebrafish is used as an experimental animal, the concentration of 2′-FL can be 500-1000 μg / mL, and the concentration of 3′-SL can be 45-2000 μg / m. It should be understood that converting the dosing dose of zebrafish to the dosing dose applicable to humans is easily achievable by those skilled in the art. For example, the theoretical human dose can be calculated according to the formula: zebrafish (mg / L) = [human (g / day) × 1000] / 6, and the actual human dose can be further deduced based on the multiple relationship between the theoretical human dose and the actual human dose, which is 0.1-4 times.

[0064] When necessary, the composition of 2′-FL and 3′-SL can also be co-administered with other active ingredients or drugs. Exemplary active ingredients or drugs for promoting brain development include, but are not limited to: cephalin, choline, PS, DHA, sialic acid, nervonic acid, etc.

[0065] Application / Method

[0066] The present invention also provides the application of the composition in the preparation of a medicament for promoting brain development.

[0067] The present invention also provides the application of the medicament in promoting brain development, and the application of the food in promoting brain development for non-therapeutic purposes.

[0068] In the present invention, "promoting brain development" can be to improve brain, nerve, and / or cognitive development, such as preventing or improving brain development retardation, nervous system development retardation, and / or cognitive development retardation. In some embodiments, "promoting brain development" includes improving cognitive maturity and promoting the expression of genes related to brain development.

[0069] In the present invention, the genes related to brain development include: bdnf, gdnfa, SOX2, DCX, GRIN2B, etc., preferably bdnf and gdnfa.

[0070] In some embodiments, "promoting brain development" is non-therapeutic and non-diagnostic.

[0071] The present invention also provides a method for promoting brain development for non-therapeutic purposes, the method comprising: administering the composition of the present invention to an individual in need, or administering the food or medicament of the present invention to an individual in need.

[0072] In the present invention, "administering" can be to introduce, provide, or deliver a certain substance to an individual through any suitable route to achieve its intended function.

[0073] The beneficial effects of the present invention at least include:

[0074] The present invention provides a composition that can promote brain development. The composition is composed of 2′-FL and 3′-SL with a mass ratio of (2.5:1) to (15:1). In this composition, 2′-FL and 3′-SL act synergistically to significantly improve the cognitive maturity of zebrafish and promote the expression of brain development-related genes bdnf and gdnfa. This composition can be used to prevent or improve retarded brain development and has good application prospects.

[0075] The following specific examples are used to further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following examples are generally carried out under conventional conditions or according to the conditions recommended by the manufacturer.

[0076] Example 1. Determination of the maximum detection concentration of 2′-FL and 3′-SL

[0077] In this example, the maximum detection concentration (MTC) of 2′-FL and 3′-SL in promoting brain development was detected respectively. The experimental method is as follows:

[0078] The zebrafish used in the experiment were all raised in fish-raising water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, the conductivity was 450 - 550 μS / cm; the pH was 6.5 - 8.5; the hardness was 50 - 100 mg / L CaCO3), and were provided by the fish-raising center of Hangzhou Huante Biotechnology Co., Ltd. Wild-type AB strain zebrafish at 5 days post-fertilization (5 dpf) were randomly selected and placed in beakers, with 30 zebrafish in each beaker (experimental group). 2′-FL or 3′-SL was administered in water (the concentrations are shown in Table 1), and a normal control group was set (no substances were added in the normal control group, only 20 mL of fish-raising water), and the capacity of each beaker was 20 mL. After treatment at 28°C for 24 h, the maximum detection concentration (MTC) of 2′-FL or 3′-SL on zebrafish was measured.

[0079] Table 1. Exploration of the efficacy concentration of 2′-FL and 3′-SL in promoting brain development (n = 30)

[0080] At the end of the experiment, no obvious abnormalities were observed in the zebrafish of the normal control group. When the treatment concentrations of 2′-FL were 125, 250, 500, and 1000 μg / mL respectively, no obvious abnormalities were observed in the zebrafish. The condition of the treatment group at the concentration of 2000 µg / mL was more serious than that of the normal control group, specifically manifested as slow movement. When the treatment concentrations of 3′-SL were 125, 250, 500, 1000, and 2000 μg / mL respectively, no obvious abnormalities were observed in the zebrafish.

[0081] The results showed that under the experimental conditions, the maximum test concentration (MTC) of 2′-FL promoting brain development was 1000 μg / mL, and the MTC of 3′-SL promoting brain development was 2000 μg / mL (Table 1).

[0082] Example 2. Study on the dose-effect relationship of 2′-FL and 3′-SL in promoting brain development

[0083] In this example, the dose-effect relationship of 2′-FL and 3′-SL in promoting brain development was explored.

[0084] In the evaluation of zebrafish brain development, color preference tests are mostly used to evaluate their color cognition. Zebrafish like short-wavelength colors. Compared with other colors (such as red, yellow, and green), zebrafish show a strong preference for blue. As the brain development is more perfect, the preference of zebrafish for blue is more obvious. The more they move in the blue area, the more mature the zebrafish's cognition is and the higher the maturity of brain development is.

[0085] The zebrafish and breeding conditions used in the experiment were the same as those in Example 1. Wild-type AB strain zebrafish at 5 dpf were randomly selected and placed in beakers, with 30 zebrafish in each beaker. 2′-FL or 3′-SL was administered by water solution (the concentrations are shown in Table 2), and the positive control was Ginkgo biloba and Cistanche deserticola tablets at a concentration of 125 μg / mL (administered by water solution, Amway (China) Co., Ltd., batch number 3313N903). At the same time, a normal control group was set up, and the capacity of each beaker was 20 mL. After treatment at 28°C for 1 day, 5 zebrafish were randomly selected from each experimental group and placed in a "cross" module. The module was divided into four areas: yellow, blue, red, and green. Six modules were placed in each group. Data were collected using a behavior analyzer, and the percentage (%) of the total movement distance of the zebrafish in the blue area within 10 min accounting for the total movement distance of the entire area was analyzed. The statistical analysis results of this index were used to evaluate the efficacy of the sample in promoting brain development. The statistical treatment results were expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software. p < 0.05 indicated that the difference was statistically significant.

[0086] Table 2. Study on the dose-effect relationship of 2′-FL and 3′-SL in promoting brain development (n = 6)

[0087] Note: Compared with the normal control group, *p < 0.05, **p < 0.01.

[0088] As shown in Table 2, when the treatment concentrations of 2′-FL were 250 μg / mL, 500 μg / mL, 750 μg / mL, and 1000 μg / mL respectively, the movement percentages of the blue area of zebrafish (p-values compared with the normal control group) were 49.7 ± 2.20% (p > 0.05), 55.2 ± 4.33% (p > 0.05), 59.4 ± 0.765% (p < 0.05), and 59.8 ± 2.88% (p < 0.05), respectively. Therefore, when the treatment concentration of 2′-FL ≥ 750 μg / mL, promoting the brain development of zebrafish has statistical significance; when the treatment concentrations of 3′-SL were 500 μg / mL, 1000 μg / mL, 1500 μg / mL, and 2000 μg / mL respectively, the movement percentages of the blue area of zebrafish (p-values compared with the normal control group) were 52.5 ± 4.10% (p > 0.05), 54.9 ± 4.52% (p > 0.05), 61.9 ± 3.71% (p < 0.05), and 63.6 ± 3.00% (p < 0.01), respectively. Therefore, when the treatment concentration of 3′-SL ≥ 1500 μg / mL, promoting the brain development of zebrafish has statistical significance.

[0089] The results show that under the experimental conditions, both 2′-FL and 3′-SL have the effect of promoting brain development, and the effective concentrations are 750 and 1500 μg / mL respectively. Specifically, compared with the normal control group, the movement percentage of the blue area of zebrafish increases, as shown in Table 2. Figure 1A and Figure 1B 。

[0090] Example 3. Efficacy study on the promotion of brain development by the composition of 2′-FL and 3′-SL

[0091] In this example, the efficacy of the composition of 2′-FL and 3′-SL in promoting brain development was explored, including its effects on the color recognition of zebrafish and the expression of brain development genes (bdnf gene, gdnfa gene).

[0092] 1. Effects of the composition of 2′-FL and 3′-SL on the color recognition of zebrafish

[0093] The zebrafish used in the experiment and the breeding conditions were the same as those in Example 1. Wild-type AB strain zebrafish at 5 dpf were randomly selected and placed in beakers, with 30 zebrafish in each beaker. A composition mixed with 2′-FL and 3′-SL at different mass percentages was administered in water (the concentrations are shown in Table 3). The positive control was Ginkgo biloba and Cistanche deserticola tablets at a concentration of 125 μg / mL (administered in water, Amway (China) Co., Ltd., batch number 3313N903). At the same time, a normal control group was set up, and the capacity of each beaker was 20 mL. After treatment at 28°C for 1 day, 5 zebrafish were randomly selected from each experimental group and placed in a "cross" module. The module was divided into four regions: yellow, blue, red, and green. Six modules were placed in each group. Data were collected using a behavior analyzer, and the percentage (%) of the total movement distance of the zebrafish in the blue region within the total movement distance of the entire region within 10 min was analyzed. The efficacy of the composition of 2′-FL and 3′-SL in promoting brain development was evaluated based on the statistical analysis results of this index. The statistical processing results were expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software, and p < 0.05 indicated that the difference was statistically significant.

[0094] Table 3. Effects of the composition of 2′-FL and 3′-SL on color recognition of zebrafish (n = 6)

[0095] Note: The concentrations of Formulas 1-7 in Table 3 are expressed as the sum of the 2′-FL concentration and the 3′-SL concentration, and the unit is μg / mL; Compared with the normal control group, *p < 0.05, **p < 0.01, ***p < 0.001; Compared with Formula 1, # p < 0.05, ## p < 0.01; Compared with Formula 2, && p < 0.01; Compared with Formula 3, @ p < 0.05, @@ p < 0.01; Compared with Formula 4, $ p < 0.05.

[0096] As can be seen from Table 3, the proportion of movement in the blue area of Formulation 1, Formulation 2, Formulation 3, Formulation 4, Formulation 5, Formulation 6, and Formulation 7 (p-values compared with the normal control group) were 62.2 ± 2.66% (p < 0.01), 67.2 ± 3.27% (p < 0.001), 62.0 ± 2.23% (p < 0.01), 61.3 ± 4.17% (p < 0.05), 50.1 ± 2.52% (p > 0.05), 54.6 ± 1.76% (p < 0.05), and 54.3 ± 2.26% (p < 0.05), respectively.

[0097] Further comparison between groups showed that Formulation 1 was statistically significant in promoting brain development. When compared with Formulation 5, Formulation 6, and Formulation 7 respectively, the p-values were p < 0.01, p < 0.05, and p < 0.05, indicating that the efficacy of Formulation 1 in promoting brain development was significantly different from that of Formulation 5, Formulation 6, and Formulation 7.

[0098] Formulation 2 was statistically significant in promoting brain development. When compared with Formulation 5, Formulation 6, and Formulation 7 respectively, the p-values were all p < 0.01, indicating that the efficacy of Formulation 2 in promoting brain development was significantly different from that of Formulation 5, Formulation 6, and Formulation 7.

[0099] Formulation 3 was statistically significant in promoting brain development. When compared with Formulation 5, Formulation 6, and Formulation 7, the p-values were p < 0.01, p < 0.05, and p < 0.05 respectively, indicating that the efficacy of Formulation 3 in promoting brain development was significantly different from that of Formulation 5, Formulation 6, and Formulation 7.

[0100] Formulation 4 was statistically significant in promoting brain development. When compared with Formulation 5, the p-value was p < 0.05, indicating that the efficacy of Formulation 4 in promoting brain development was significantly different from that of Formulation 5.

[0101] The above results show that under the conditions of this experiment, compared with the normal control group, the increased proportions of movement in the blue area of Formulation 1 - 4 were 16.3%, 21.3%, 16.1%, and 15.4% respectively, all showing the efficacy of promoting brain development, which was better than the positive control Ginkgo biloba and Cistanche deserticola tablets. Moreover, the effects of Formulation 1 - 3 were better than those of Formulation 5 - 7, and the effect of Formulation 4 was better than that of Formulation 5, indicating that when 2′-FL and 3′-SL in Formulation 1 - 4 were in the range of (2.5:1) - (15:1), they had a synergistic effect on promoting brain development. See Table 3 for details. Figure 2A and Figure 2B 。

[0102] 2. Effects of the composition of 2′-FL and 3′-SL on gene expression related to brain development

[0103] In this example, the bdnf gene and the gdnfa gene were used as representative genes for brain development to explore the effect of the combination of 2′-FL and 3′-SL on the expression of brain development genes.

[0104] Bdnf and its receptor are widely expressed in the nervous system, with the highest content in the hippocampus and cortex. Their modes of action in the central nervous system are as follows: (1) increasing synaptic plasticity, which in turn affects long-term potentiation (LTP), the basis of the learning process and memory formation (secondary memory); (2) promoting neurogenesis, especially in the hippocampus; (3) promoting cell survival, mainly maintaining and promoting the development, differentiation, growth, and regeneration of various neurons, especially 5-hydroxytryptamine (5-HT) and dopamine (DA) neurons. Gdnfa can promote the survival of different neuron subsets at different stages of central and peripheral nervous system development, support the generation of type 1 astrocytes, Schwann cells, neurons, pinealocytes, etc., and has a significant effect on promoting the survival of spinal motor neurons. Therefore, upregulation of the expression levels of these two genes can promote central nervous system and nerve development, and thus promote the formation of learning and memory, making them representative genes for brain development.

[0105] The zebrafish used in the experiment and the breeding conditions were the same as those in Example 1. Wild-type AB strain zebrafish at 5 dpf were randomly selected and placed in beakers, with 30 zebrafish in each beaker (experimental group). The samples were administered in water (concentrations are shown in Tables 4 and 5), and the positive control was Ginkgo biloba and Cistanche deserticola tablets at a concentration of 125 μg / mL. At the same time, a normal control group was set up, and the volume of each beaker was 20 mL. The experiments were set up in parallel three times. After treatment at 28°C for 1 day, total RNA of zebrafish in each group was extracted using a pre-installed magnetic bead method universal RNA extraction kit (product number TL2402001643C, ONREW, China), and the concentration and purity of the total RNA were measured using an ultraviolet-visible spectrophotometer. 2.00 μg of total RNA from zebrafish samples was taken, and 20.0 μL of cDNA was synthesized according to the instructions of the cDNA first-strand synthesis kit (batch number: H9305270, Yeasen Biotechnology (Shanghai) Co., Ltd., China). The expression of β-actin, bdnf, and gdnfa genes was detected by q-PCR. β-actin was used as an internal reference for gene expression, and the relative expression levels of bdnf and gdnfa genes were calculated. The statistical treatment results were expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software, and p < 0.05 indicated that the difference was statistically significant.

[0106] (1) Effect of the combination of 2′-FL and 3′-SL on the expression of the bdnf gene

[0107] Table 4. Effects of the combination of 2′-FL and 3′-SL on the expression of the bdnf gene (n = 3)

[0108] Note: The concentrations of Formulas 1-7 in Table 4 are expressed as the concentration of 2′-FL + the concentration of 3′-SL, and the unit is μg / mL; Compared with the normal control group, **p < 0.01, ***p < 0.001; Compared with Formula 1, # p < 0.05, ### p < 0.001; Compared with Formula 2, & p < 0.05, && p < 0.01, &&& p < 0.001; Compared with Formula 3, @ p < 0.05, @@ p < 0.01, @@@ p < 0.001; Compared with Formula 4, $ p < 0.05, $$$ p < 0.001.

[0109] As can be seen from Table 4, the relative expression levels of the bdnf gene (p values compared with the normal control group) of Formulas 1, 2, 3, 4, 5, 6, and 7 are 5.51 ± 0.117 (p < 0.001), 2.65 ± 0.194 (p < 0.01), 4.26 ± 0.265 (p < 0.001), 3.73 ± 0.161 (p < 0.001), 1.01 ± 0.065 (p > 0.05), 1.56 ± 0.046 (p < 0.001), and 1.48 ± 0.056 (p < 0.01), respectively. Therefore, compared with the blank control group, Formulas 1, 2, 3, 4, 6, and 7 have statistical significance in upregulating the relative expression level of the bdnf gene; Formula 5 has no statistical significance in upregulating the relative expression level of the bdnf gene.

[0110] The p values of Formula 1 for upregulating the relative expression level of the bdnf gene compared with Formulas 5, 6, and 7 are all p < 0.001, indicating that the efficacy of Formula 1 in upregulating the relative expression level of the bdnf gene is statistically significant compared with Formulas 5, 6, and 7.

[0111] For Formula 2 in upregulating the relative expression level of the bdnf gene, the p-values compared with Formulas 5, 6, and 7 were p < 0.01, p < 0.01, and p < 0.01 respectively, indicating that the efficacy of Formula 2 in upregulating the relative expression level of the bdnf gene was statistically significant compared with Formulas 5, 6, and 7.

[0112] For Formula 3 in upregulating the relative expression level of the bdnf gene, the p-values compared with Formulas 5, 6, and 7 were all p < 0.001, indicating that the efficacy of Formula 3 in upregulating the relative expression level of the bdnf gene was statistically significant compared with Formulas 5, 6, and 7.

[0113] For Formula 4 in upregulating the relative expression level of the bdnf gene, it was statistically significant. The p-values compared with Formulas 5, 6, and 7 were all p < 0.001, indicating that the efficacy of Formula 4 in upregulating the relative expression level of the bdnf gene had a synergistic effect compared with Formulas 5, 6, and 7.

[0114] The above results showed that under the conditions of this experiment, compared with the normal control group, the increments of the relative expression level of the bdnf gene for Formulas 1 - 4 were 4.51, 1.65, 3.26, and 2.73 respectively, all showing promotion of brain development, being superior to the positive control Ginkgo biloba and Cistanche deserticola tablets, and all being superior to the effects of Formulas 5 - 7. It indicated that when 2′-FL and 3′-SL in Formulas 1 - 4 were in the range of (2.5:1) - (15:1), they had a synergistic effect on promoting brain development. See Table 4 and Figure 3 。

[0115] (2)Effect of the composition of 2′-FL and 3′-SL on the expression of the gdnfa gene

[0116] Table 5. Effect of the composition of 2′-FL and 3′-SL on the expression of the gdnfa gene (n = 3)

[0117] Note: The concentrations of Formulas 1 - 7 in Table 5 are expressed as the 2′-FL concentration + 3′-SL concentration, and the unit is μg / mL; Compared with the normal control group, **p < 0.01, ***p < 0.001; Compared with Formula 1, # p < 0.05, ## p < 0.01; Compared with Formula 2, & p < 0.05, && p < 0.01, &&& p < 0.001; Compared with Formula 3, @p < 0.05, @@ p < 0.01, @@@ p < 0.001; Compared with Formula 4, $ p < 0.05, $$ p < 0.01.

[0118] As can be seen from Table 5, the relative expression levels of the gdnfa gene in Formulas 1, 2, 3, 4, 5, 6, and 7 (p-values compared with the normal control group) were 1.83 ± 0.062 (p < 0.001), 2.42 ± 0.098 (p < 0.001), 2.49 ± 0.143 (p < 0.001), 2.28 ± 0.136 (p < 0.001), 1.14 ± 0.060 (p > 0.05), 1.29 ± 0.027 (p < 0.01), and 1.58 ± 0.083 (p < 0.01), respectively. Therefore, Formulas 1, 2, 3, 4, 6, and 7 are statistically significant in upregulating the relative expression level of the gdnfa gene; Formula 5 is not statistically significant in upregulating the relative expression level of the gdnfa gene.

[0119] Formula 1 is statistically significant in upregulating the relative expression level of the gdnfa gene. The p-values compared with Formulas 5 and 6 are p < 0.01 and p < 0.01, respectively, indicating that the efficacy of Formula 1 in upregulating the relative expression level of the gdnfa gene is statistically significant compared with Formulas 5 and 6.

[0120] Formula 2 is statistically significant in upregulating the relative expression level of the gdnfa gene. The p-values compared with Formulas 5, 6, and 7 are p < 0.001, p < 0.001, and p < 0.01, respectively, indicating that the efficacy of Formula 2 in upregulating the relative expression level of the gdnfa gene is statistically significant compared with Formulas 5, 6, and 7.

[0121] Formula 3 is statistically significant in upregulating the relative expression level of the gdnfa gene. The p-values compared with Formulas 5, 6, and 7 are p < 0.001, p < 0.01, and p < 0.01, respectively, indicating that the efficacy of Formula 3 in upregulating the relative expression level of the gdnfa gene is statistically significant compared with Formulas 5, 6, and 7;

[0122] Formula 4 is statistically significant in upregulating the relative expression level of the gdnfa gene. The p-values compared with Formulas 5, 6, and 7 are p < 0.01, p < 0.01, and p < 0.05, respectively, indicating that the efficacy of Formula 4 in upregulating the relative expression level of the gdnfa gene is statistically significant compared with Formulas 5, 6, and 7;

[0123] The above results show that under the conditions of this experiment, compared with the normal control group, the relative expression value increments of the gdnfa gene in Formulas 1-4 are 0.83, 1.42, 1.49, and 1.28 respectively, all of which show promotion of brain development, superior to the positive control Ginkgo biloba and Cistanche deserticola tablets, and all are superior to the effects of Formulas 5-7. It shows that when 2′-FL and 3′-SL in Formulas 1-4 are in the range of (2.5:1) to (15:1), they have a synergistic effect on promoting brain development. See Table 5 and Figure 4 .

[0124] Example 4. Comparison of the composition of 2′-FL and 3′-SL with other compositions

[0125] During the research process, through repeated experiments, the inventor found that only the composition of 2′-FL and 3′-SL has a synergistic effect on promoting brain development when in a specific ratio range of (2.5:1) to (15:1), while the composition with 3-FL added on the basis of 2′-FL and 3′-SL and the composition not in this ratio range cannot play a good role in promoting brain development.

[0126] In this example, a composition with a ratio of 2′-FL and 3′-SL of 9:5.89 (i.e., a 2-component ratio group), and a composition with a ratio of 2′-FL, 3-FL and 3′-SL of 9:1:5.89 (i.e., a 3-component ratio group) are used as comparisons, and a composition with a ratio of 2′-FL and 3′-SL of 7:1 (i.e., the optimal ratio group) is used as an example of the composition of the present invention to compare the differences in promoting brain development among these three compositions.

[0127] The zebrafish and breeding conditions used in the experiment are the same as in Example 1. Randomly select 5 dpf wild-type AB strain zebrafish into a beaker, and each beaker (experimental group) is treated with 30 zebrafish. Administer the composition of 2′-FL and 3′-SL or other compositions (concentrations are shown in Table 6) by water solution, with the positive control Ginkgo biloba and Cistanche deserticola tablets at a concentration of 125 μg / mL. At the same time, set up a normal control group, and the capacity of each beaker is 20 mL. After treatment at 28°C for 24 h, randomly select 5 zebrafish from each experimental group and put them into a "cross" module. The module is divided into four regions: yellow, blue, red, and green. Six modules are placed in each group. Use a behavior analyzer to collect data and analyze the percentage (%) of the total movement distance of the zebrafish in the blue region within the total movement distance of the entire region within 10 min. Evaluate the efficacy of the sample in promoting brain development based on the statistical analysis results of this index. The statistical processing results are expressed as mean±SE. Use SPSS 26.0 software for statistical analysis, and p < 0.05 indicates that the difference is statistically significant.

[0128] Table 6. Comparison of the effects of the compositions of 2′-FL and 3′-SL with other compositions on promoting brain development (n = 6)

[0129] Note: Compared with the normal control group, ***p < 0.001; compared with the group with the ratio of three raw materials, #p < 0.05

[0130] As can be seen from Table 6, Figure 5A and Figure 5B the proportions of movement in the blue area of the optimal ratio group, the group with the ratio of two raw materials, and the group with the ratio of three raw materials are 64.8 ± 1.59%, 58.8 ± 1.83%, and 58.5 ± 1.63% respectively. When compared with the normal control group (45.4 ± 2.10%), the p-values are all p < 0.001. Therefore, the optimal ratio group, the group with the ratio of two raw materials, and the group with the ratio of three raw materials all have statistical significance in promoting brain development.

[0131] Further statistics show that the p-value when the optimal ratio group is compared with the group with the ratio of three raw materials is p < 0.05, indicating that the optimal ratio group has a better effect on promoting brain development than the group with the ratio of three raw materials. When the optimal ratio group, the group with the ratio of two raw materials, and the group with the ratio of three raw materials are respectively compared with the positive control Ginkgo biloba and Cistanche deserticola tablets, only the optimal ratio group is better than the positive control Ginkgo biloba and Cistanche deserticola tablets, indicating that the composition of the present invention is more excellent in increasing the color recognition of zebrafish and has a very significant function of promoting brain development.

[0132] When the group with the ratio of two raw materials is compared with the group with the ratio of three raw materials, the result shows that the p-value of the comparison between the group with the ratio of two raw materials and the group with the ratio of three raw materials is p > 0.05, indicating that there is no statistical difference in the effect of promoting brain development between the group with the ratio of two raw materials and the group with the ratio of three raw materials, which means that adding 3-FL on the basis of 2′-FL and 3′-SL cannot further improve the function of the composition in promoting brain development.

[0133] The above embodiments only represent several implementation manners of the present invention. The description is relatively specific and detailed, but it should not be construed as a limitation on the scope of the patent of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the appended claims. At the same time, all the documents mentioned in the present invention are cited in this application as references, just as each document is cited separately as a reference.

Claims

1. A composition, which is composed of 2′-FL and 3′-SL with a mass ratio of (2.5:1) to (15:1).

2. The composition according to claim 1, wherein The amount of 2′-FL is 2.5 to 10 parts by mass relative to 1 part by mass of 3′-SL.

3. A food product, characterized in that, The food contains the composition according to claim 1 or 2.

4. The food according to claim 3, characterized in that, The food includes finished food products, semi-finished food products, food additives, and food supplements.

5. The food according to claim 3, characterized in that, The food further includes pharmaceutically acceptable materials.

6. The food according to claim 5, characterized in that, The pharmaceutically acceptable materials include: nutritional additives, ingredients of both food and medicine, excipients, and / or auxiliary materials.

7. The food according to claim 6, wherein The nutritional additives include one or more of dietary fiber, prebiotics, proteins, lipids, minerals, and vitamins, and / or the ingredients of both food and medicine include one or more of red dates, hawthorn, wolfberries, longans, lilies, poria cocos, and dried tangerine peels, and / or the excipients or auxiliary materials include one or more of calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.

8. The food according to any one of claims 3-7, characterized in that, The food is infant food, children's food, juvenile food, adolescent food, young adult food, adult food, middle-aged food, or elderly food.

9. The food according to claim 8, characterized in that, The infants include babies, older infants, and toddlers.

10. The food according to claim 8, characterized in that, The food is infant formula milk powder, baby complementary food, children's formula milk powder, children's snacks, pregnant women's formulated milk powder, middle-aged and elderly milk powder, or nutritional or dietary supplements.

11. A drug, characterized in that, The drug contains an effective amount of a composition composed of 2′-FL and 3′-SL with a mass ratio of (2.5:1) to (15:1), and a pharmaceutically acceptable carrier.

12. The drug according to claim 11, characterized in that, The amount of 2′-FL is 2.5 to 10 parts by mass relative to 1 part by mass of 3′-SL.

13. The drug according to claim 11, characterized in that, The effective amount of 2′-FL is 500 to 1000 μg / mL, and / or the effective amount of 3′-SL is 45 to 2000 μg / mL.

14. Use of the composition according to claim 1 or 2 in the preparation of a drug for promoting brain development.

15. Use of the drug according to any one of claims 11 - 13 in promoting brain development, or use of the food according to any one of claims 3 - 10 in promoting brain development for non-therapeutic purposes.

16. The application according to claim 14 or 15, characterized in that, The promotion of brain development is to improve brain, nerve, and / or cognitive development.

17. The application according to claim 16, characterized in that, The promotion of brain development includes preventing or improving brain development retardation, nervous system development retardation, and / or cognitive development retardation.

18. The application according to claim 16, characterized in that, The promotion of brain development includes improving cognitive maturity and promoting the expression of genes related to brain development.

19. The application according to claim 18, wherein The brain development-related genes include bdnf, gdnfa.

20. A method for promoting brain development for non-therapeutic purposes, characterized in that, The method includes: administering the composition according to claim 1 or 2 to an individual in need, or administering the food according to any one of claims 3 - 10 or the drug according to any one of claims 11 - 13 to an individual in need.

21. The method according to claim 20, characterized in that, The promotion of brain development is as defined in any one of claims 16 - 19.

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