Composition for promoting infant brain development and application thereof

Abstract

The invention belongs to the technical field of microbial engineering, discloses a composition for promoting infant brain development and application thereof, and provides a composition containing lactobacillus paracasei N1115, breast milk oligosaccharide and whey protein powder, the composition can promote infant brain and nerve development, all the components have a synergistic effect, and the effect of promoting infant brain and nerve development is achieved. The conversion from latent TGF-beta in the whey protein powder to activated TGF-beta is activated, so that the aims of promoting neurodevelopment, improving memory and promoting brain development of infants are fulfilled. The preparation method is suitable for preparing milk powder, cheese, yoghourt, plant-based yoghourt and derivative products or other foods for promoting brain development of infants.

Description

Technical Field

[0001] This invention belongs to the field of nutritional composition technology, specifically a composition for promoting brain development in infants and young children and its application. Background Technology

[0002] Infancy is a critical stage for brain development, and the nutritional supply during this period is essential for the formation of brain structure, the proliferation and differentiation of nerve cells, and the development of cognitive abilities. Therefore, developing nutritional compositions that can effectively promote brain development in infants and young children is of great significance.

[0003] Among the many probiotics related to infant health, *Lactobacillus paracasei* is one of the important beneficial probiotics for infants. Studies have shown that it indirectly supports infant brain development through multiple pathways, such as promoting the absorption of nutrients needed for brain development in the gut.

[0004] In infant brain development, TGF-β, a component of Transforming Growth Factor (TGF), plays a crucial role, actively regulating nerve cell growth, migration, and synapse formation. Due to its small molecular weight, TGF cannot be directly added to nutritional supplements; instead, substances rich in TGF are typically added. Whey protein powder, as a high-quality protein source, not only provides essential amino acids and other nutrients necessary for brain development in infants and young children but also contains TGF. However, in whey protein powder, TGF often exists in a latent form, TGF-β, which has no biological functional activity.

[0005] Latent TGF-β has significant limitations in promoting brain development: because it cannot bind to TGF-β receptors, it cannot activate downstream signaling pathways related to brain development, making it difficult to exert its regulatory effects on nerve cell growth, synapse formation, etc. This means that the beneficial TGF-β in whey protein powder for brain development cannot be fully utilized, thus affecting its efficacy in promoting brain development in infants and young children.

[0006] Given the specific nutritional support requirements for infant brain development and the low utilization rate of TGF-β, a beneficial component in existing whey protein powders, finding probiotics that can activate latent TGF-β in whey protein powders has become a technical challenge in the development of probiotic products. Summary of the Invention

[0007] To address the aforementioned problems in existing technologies, our team conducted in-depth research and discovered that even strains of the same genus and species exhibit significant differences in their efficacy in promoting infant brain development. Among numerous strains, the specific strain *Lactobacillus paracasei* N1115 can synergistically promote infant brain development with human milk oligosaccharide compositions. It can also efficiently and specifically activate the conversion of latent TGF-β in whey protein powder into activated TGF-β, thereby solving the problem that directly using whey protein powder in nutritional compositions for promoting infant brain development cannot fully utilize its TGF-β, hindering nutrient absorption and utilization, and resulting in poor effects on promoting infant brain development.

[0008] The present invention also provides a composition containing Lactobacillus paracasei N1115 to promote brain development in infants and young children. The components in this composition have a synergistic effect, which can achieve the purpose of promoting brain development in infants and young children.

[0009] The present invention also provides the use of compositions that promote brain development in infants and young children in the preparation of probiotic powders, oil drops, infant formula or children's formula that have the effect of promoting brain development in infants and young children.

[0010] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0011] A composition for promoting brain development in infants and young children, comprising Lactobacillus paracasei N1115, human milk oligosaccharides (HMOs), and whey protein.

[0012] The Lactobacillus paracasei strain N1115, with the Latin name Lactobacillus paracasei, was isolated from traditional fermented dairy products in Inner Mongolia. It was deposited at the China General Microbiological Culture Collection Center (CGMCC) on March 17, 2011, with accession number CGMCC No. 4691, and was published in Chinese Patent Application No. CN201110357058.8 on January 23, 2013.

[0013] As a limitation of the present invention, the composition comprises a dosage ratio of 1×10 11 ~1×10 12 CFU: 5-30g: 15-50g of Lactobacillus paracasei N1115, human milk oligosaccharides, and whey protein.

[0014] As a further limitation of the present invention, the viable count of *Lactobacillus paracasei* N1115 in the composition is ≥1×10⁻⁶. 7 CFU / g.

[0015] As a further limitation of the present invention, the *Lactobacillus paracasei* N1115 is used to activate latent transforming growth factor-β in whey protein powder.

[0016] As a further limitation of the invention, the human milk oligosaccharide is a mixture of 3'-sialyl lactose (i.e., 3'SL) and 6'-sialyl lactose (i.e., 6'SL).

[0017] As a further limitation of the present invention, the weights of 3'SL and 6'SL in the human milk oligosaccharide are equal.

[0018] The present invention also provides an application of a composition for promoting brain development in infants and young children, wherein the composition for promoting brain development in infants and young children is used to prepare probiotic agents or health foods.

[0019] As a further limitation of the present invention, the probiotic agent is a probiotic powder or oil drop.

[0020] As a further definition of the present invention, the health food is milk powder, cheese, yogurt or plant-based yogurt that promotes brain development in infants and young children.

[0021] As a further limitation of the present invention, the composition for promoting brain development in infants and young children is added in the amount of 1.5% to 10% by weight in infant formula, children's formula or health food.

[0022] By adopting the above-described technical solution, the beneficial effects achieved by this invention compared to the prior art are as follows:

[0023] (1) This invention discovered that the combination of Lactobacillus paracasei N1115 and human milk oligosaccharides has a synergistic effect on promoting brain and nerve development in infants and young children. It was also unexpectedly discovered that Lactobacillus paracasei N1115 can relieve the inhibition of latent TGF-β activity in whey protein powder and promote the conversion of latent TGF-β to activated TGF-β.

[0024] TGF-β conversion provides a new technological approach to promoting brain and nerve development in infants and young children.

[0025] (2) The two components of the present invention, Lactobacillus paracasei N1115 and whey protein powder, work synergistically to promote brain development in infants and young children. When used in combination, they not only activate the transforming growth factor in whey protein powder that regulates immune function, but also significantly improve spatial memory and learning and cognitive abilities compared to using Lactobacillus paracasei N1115 or whey protein powder alone, thus promoting brain development.

[0026] (3) The composition of the present invention that promotes brain development in infants and young children can be applied to probiotic powder, milk powder, cheese, yogurt or plant-based yogurt, and can promote intestinal flora health and significantly increase the abundance of Lactobacillus and Bifidobacterium.

[0027] This invention is applicable to the preparation of compositions that promote brain development in infants and young children, and is used to prepare infant formula or children's formula that promotes brain development in infants and young children. Attached Figure Description

[0028] The present invention will now be described in further detail with reference to the accompanying drawings and specific embodiments.

[0029] Figure 1 This is a diagram showing the relative abundance of gut microbiota at the phylum level in each experimental group during the verification experiment of this invention.

[0030] Figure 2 This is a graph showing the relative abundance of gut microbiota at the genus level in each experimental group during the verification experiment of this invention. Detailed Implementation

[0031] The present invention will be further described in detail below with reference to specific embodiments and accompanying drawings. It should be understood that the described embodiments are only used to explain the present invention and do not limit the present invention.

[0032] Unless otherwise specified, all materials and reagents used in the embodiments of this invention are commercially available. Experimental methods not specifically described in the embodiments are generally performed under standard conditions or as recommended by the manufacturer.

[0033] In the verification experiments of this invention, the reagent dosages were all verified through minimum toxic concentration detection experiments. Among them, *Lactobacillus paracasei* JMCC0012 is deposited at the Junlebao Lactobacillus Culture Collection Center.

[0034] Example 1: A composition for promoting brain development in infants and young children

[0035] This embodiment describes a composition for promoting brain development in infants and young children. The composition consists of components in a dosage ratio of 1×102 12 This product is made from a mixture of CFU:5g:15g of Lactobacillus paracasei N1115, human milk oligosaccharides, and whey protein powder. The human milk oligosaccharides are prepared by mixing equal weights of 3'SL and 6'SL.

[0036] This composition, which promotes brain development in infants and young children, is used to prepare probiotic powder, probiotic oil drops, infant formula, or children's formula, wherein the viable count of *Lactobacillus paracasei* N1115 is ≥1×10⁻⁶. 7 CFU / g.

[0037] It has been verified that *Lactobacillus paracasei* N1115 and the above-mentioned composition promoting infant brain development can be used to activate latent TGF-β in whey protein powder to convert it into activated TGF-β, which has the function of promoting infant brain development. The specific verification experiments are as follows:

[0038] (I) Activation verification experiment of latent TGF-β

[0039] (1.1) Group settings:

[0040] Experimental group 1: 10.5 ml of MRS culture medium served as a blank control;

[0041] Experimental Group 2: 0.5 mL of activated *Lactobacillus paracasei* JMCC0012 bacterial suspension was placed in 10 mL of MRS medium and incubated statically at 37°C for 4 h, equivalent to 1 × 10⁻⁶ bacteria containing *Lactobacillus paracasei* JMCC0012. 11 CFU;

[0042] Experimental group 3: 0.5 g of a human milk oligosaccharide mixture prepared by mixing equal weights of 3'SL and 6'SL was added to 10 ml LMR S medium;

[0043] Experimental Group 4: 0.5 mL of activated *Lactobacillus paracasei* N1115 bacterial suspension was placed in 10 mL of MRSE medium and incubated at 37°C for 4 h; this is equivalent to 1 × 10⁻⁶ *Lactobacillus paracasei* N1115. 11 CFU;

[0044] Experimental group 5: 1.5g of concentrated whey protein powder was added to 10ml of LMR S medium;

[0045] Experimental group 6: 0.5g of HMOs prepared by mixing equal weights of 3'SL and 6'SL and 1.5g of concentrated whey protein powder were placed in 10ml of MRS medium and co-cultured at 37℃ for 4h.

[0046] Experimental group 7: 0.5 mL of activated Lactobacillus paracasei N1115 bacterial culture and 1.5 g of concentrated whey protein powder were placed in 10 mL of MRS medium and co-cultured at 37 °C for 4 h.

[0047] Experimental group 8: 0.5 mL of activated Lactobacillus paracasei JMCC0012 bacterial culture and 1.5 g of concentrated whey protein powder were placed in 10 mL of MRS medium and co-cultured at 37 °C for 4 h.

[0048] Experimental group 9: 0.5g of HMOs prepared by mixing equal weights of 3'SL and 6'SL and 0.5mL of activated Lactobacillus paracasei N1115 bacterial suspension were placed in 10ml of MRS medium and co-cultured at 37℃ for 4h.

[0049] Experimental group 10: 0.5g of HMOs prepared by mixing equal weights of 3'SL and 6'SL, 0.5mL of activated Lactobacillus paracasei N1115 bacterial culture, and 1.5g of concentrated whey protein powder were placed in 10ml of MRS medium and co-cultured at 37℃ for 4h.

[0050] Experimental group 11: 0.5g of HMOs prepared by mixing equal weights of 3'SL and 6'SL, 0.5mL of activated Lactobacillus paracasei JMCC0012 and 1.5g of concentrated whey protein powder were placed in 10ml of MRS medium and co-cultured at 37℃ for 4h.

[0051] The bacterial content in each of the above groups was equivalent to 1×10⁶. 11 CFU.

[0052] (1.2) Detection method:

[0053] After the culture was completed, equal amounts of culture medium samples from each experimental group were taken, and the contents of latent and activated TGF-β were quantitatively detected by enzyme-linked immunosorbent assay (ELISA). The results are shown in Table 1.

[0054] Table 1. Experimental results for verifying the activation of latent TGF-β

[0055]

[0056]

[0057] The results show that the proportion of activated TGF-β in concentrated whey protein powder is 2.25%. The human milk oligosaccharide composition containing *Lactobacillus paracasei* N1115 can increase the proportion of activated TGF-β in concentrated whey protein powder to 42.3%. However, JMCC0012, which belongs to the same genus and species as N1115, has a relatively weak activation effect on latent TGF-β and cannot synergistically promote TGF-β activation. Therefore, *Lactobacillus paracasei* N1115 can improve the nutrient utilization efficiency of whey protein in the composition product.

[0058] (II) Verification Experiment on Improving Memory and Cognitive Learning Abilities

[0059] This invention verifies, through Y-maze experiments and novel object experiments, that the composition of Embodiment 1 of the present invention for promoting infant brain development has the function of improving memory and cognitive learning ability, thereby promoting brain nerve development and infant brain development.

[0060] The specific experimental verification process is as follows:

[0061] Newborn mice were randomly divided into groups of equal size. Each group was administered the same number of mice via gavage according to Table 2, while simultaneously receiving the same amount of feed as usual. After 21 days of continuous intervention, the Y-maze test and the new object test were performed, and the mice's feces were collected to detect their intestinal flora.

[0062] Table 2. Experimental group intervention methods

[0063]

[0064]

[0065] (1) Y-maze experiment

[0066] The Y-maze consists of three arms of equal length, forming a 120° angle between them. Each arm is approximately 10cm wide and 40cm long. A mouse is placed in the central area of ​​the Y-maze, facing one of the arms, and allowed to freely explore the maze for 5–10 minutes. The number of times the mouse enters each arm and the sequence of arms entered are recorded. The spontaneous alternation rate is calculated; a higher spontaneous alternation rate indicates better spatial memory. A higher number of spontaneous alternations also indicates better spatial memory.

[0067] (2) New Object Experiment

[0068] The new object experiment is divided into three phases: adaptation, familiarization, and testing. During the adaptation phase, mice are placed alone in a box without any objects and explore freely for 5 minutes. During the familiarization phase, two identical rectangular objects (6.4cm long, 6.2cm wide, and 12.5cm high) are fixed 10cm from the sides of the box. Mice enter the box alone with their backs to the objects and explore them for 10 minutes. During the testing phase, one of the objects is replaced with a different shaped object (5cm long, 2.5cm wide, and 10cm high). The exploration criteria are the same as during the familiarization phase, and the experiment lasts for 10 minutes.

[0069] The longer the exploration time for a new object, the better the mouse's spatial memory ability. Assess the animal's learning and memory abilities. Calculate the mouse cognitive index (RI): RI = (Exploration time for new object / (Exploration time for new object + Exploration time for old object)) × 100%. This reflects the animal's preference for new objects; the higher the RI value, the stronger the mouse's learning and memory ability.

[0070] The results of the Y-maze experiment and the new object experiment are shown in Table 3:

[0071] Table 3 Results of the Y-maze experiment and the new object experiment

[0072]

[0073]

[0074] Note: The letters a, b, and c represent significant differences between the two experimental groups, p < 0.05.

[0075] The results of the Y-maze experiment and the new object experiment show that the composition of human milk oligosaccharides and concentrated whey protein powder containing Lactobacillus paracasei N1115 of the present invention can improve the memory and cognitive learning ability of mice, and thus can be used to promote the neural and brain development of infants and young children. However, JMCC0012, which belongs to the same genus and species as N1115, does not have a significant synergistic effect with human milk oligosaccharides and concentrated whey protein powder in promoting brain development.

[0076] (3) Intestinal flora detection experiment

[0077] The relative abundance of gut microbiota at the phylum and genus levels in each group of mice was measured, and a percentage stacked bar chart was plotted. Figure 1 and Figure 2 .

[0078] Depend on Figure 1 and Figure 2 It can be seen that, among the phylum-level detection results, the abundance of Armatimonadetes, Acidobacteria, Bacteroidetes, Fusobacteria, Gemmatimonadetes, Deinococcus-Thermus, Candidatus-Saccharibacteria, Proteobacteria, Actinobacteria, Nitrospirae, Chloroflexi, Firmicutes, Verrucomicrobia, and Deferribacteres was relatively high.

[0079] Among the genera detected, the abundance of Bifidobacterium, Lactobacillus, Faecalibacterium, Blautia, Ruminococcus, Collinsella, Bacteroides, Anaerostipes, Enterococcus, Streptococcus, Eubacterium, Roseburia, Alistipes, and Akkermansia was relatively high.

[0080] The results showed that, at the phylum level, the combination of N1115, HMOs, and concentrated whey protein powder significantly increased the abundance of Firmicutes compared to other groups. At the genus level, the compositions of this invention significantly increased the abundance of Lactobacillus and Bifidobacterium, thereby promoting a healthy gut environment.

[0081] Examples 3-6: Compositions for Promoting Brain Development in Infants and Young Children

[0082] Examples 3-6 are compositions for promoting brain development in infants and young children, and their specific components and dosages are shown in Table 5.

[0083] Table 5 shows the specific components and dosages of each composition in Examples 3-6.

[0084]

[0085] Verification showed that the viable count of *Lactobacillus paracasei* N1115 in each composition was ≥1×10⁻⁶. 7 CFU / g, all of the above compositions can be used to promote brain development in infants and young children.

[0086] Examples 7-10: Infant formula milk powder containing compositions that promote brain development in infants and young children

[0087] Examples 7-10 are infant formula milk powders containing compositions that promote brain development in infants and young children.

[0088] The infant formula in Example 7 contains 5.5% by weight of the composition from Example 1 that promotes brain development in infants and young children.

[0089] The infant formula in Example 8 contains 10% by weight of the composition from Example 2 that promotes brain development in infants.

[0090] The infant formula in Example 9 contains 7.5% by weight of the composition from Example 3 that promotes brain development in infants.

[0091] The infant formula in Example 10 contains 1.5% by weight of the composition from Example 4 that promotes brain development in infants and young children.

[0092] In other embodiments, the composition for promoting infant brain development is used to prepare probiotic powders, oil drops, cheese, yogurt, or plant-based yogurt, wherein the viable count of *Lactobacillus paracasei* N1115 in the composition is ≥1×10⁻⁶. 7 CFU / g.

[0093] It should be noted that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art can still modify the technical solutions described in the above embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. A composition for promoting brain development in infants and young children, characterized in that, The composition contains Lactobacillus paracasei N1115, human milk oligosaccharides, and whey protein.

2. The composition for promoting infant brain development according to claim 1, characterized in that, The composition contains a dosage ratio of 1×10 11 ~1×10 12 CFU: 5~30g: 15~50g of Lactobacillus paracasei N1115, human milk oligosaccharides, and whey protein.

3. The composition for promoting brain development in infants and young children according to claim 2, characterized in that, The viable count of *Lactobacillus paracasei* N1115 in the composition is ≥1×10⁻⁶. 7 CFU / g.

4. The composition for promoting brain development in infants and young children according to claim 3, characterized in that, The Lactobacillus paracasei N1115 is used to activate latent transforming growth factor-β in whey protein powder.

5. The composition for promoting brain development in infants and young children according to claim 4, characterized in that, The human milk oligosaccharide is a mixture of 3'-sialyl lactose and 6'-sialyl lactose.

6. The composition for promoting brain development in infants and young children according to claim 5, characterized in that, The human milk oligosaccharide contains 3'-sialyl lactose and 6'-sialyl lactose in equal weights.

7. The application of a composition for promoting brain development in infants and young children according to any one of claims 1 to 6, characterized in that, The composition that promotes brain development in infants and young children is used to prepare probiotics or health foods.

8. The application of the composition for promoting infant brain development according to claim 7, characterized in that, The probiotic agent is a probiotic powder or oil drop.

9. The application of the composition for promoting brain development in infants and young children according to claim 7, characterized in that, The health food products mentioned are milk powder, cheese, yogurt, or plant-based yogurt that promote brain development in infants and young children.

10. The application of the composition for promoting brain development in infants and young children according to claim 9, characterized in that, The amount of the composition for promoting infant brain development added to milk powder is 1.5% to 10% by weight.

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