Human milk oligosaccharides for early intervention to enhance memory capacity in adulthood and uses thereof

Abstract

The present application provides a breast milk oligosaccharide for early intervention to improve the memory ability of an individual in adulthood and an application thereof. Specifically, the present application provides an application of a breast milk oligosaccharide as an active component in the preparation of a product for helping to improve and / or improve the memory ability of an individual in adulthood; wherein the breast milk oligosaccharide is selected from one or more of 2'-fucosyllactose (2'-FL), 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL); the product is an oral product for an individual in the pre-adult period. The present application can positively promote the cognitive development, especially the memory ability, in adulthood by early intervention of breast milk oligosaccharide.

Description

Technical Field

[0001] This invention relates to the field of functional nutrient research technology, specifically to a human milk oligosaccharide that helps enhance and / or improve an individual's memory ability in adulthood and its applications. Background Technology

[0002] The first 1000 days of human life are the golden period for brain development and the period of most rapid cognitive development in infants and young children, having a profound impact on an individual's long-term health and intellectual development. During this period, the brain undergoes developmental processes such as neural tube formation, rapid neuronal proliferation, nerve cell migration and differentiation, synapse formation and pruning, and neuronal myelination, ultimately forming a basically mature structure. These processes are crucial for the subsequent establishment of neural networks and the exercise of higher neural functions (such as visual information processing, language learning, emotional responses, cognition and consciousness, and social interaction). Nutrition plays a vital role in this process. Nutritional deficiencies, imbalances, or exposure to unhealthy dietary factors during this period (such as high-fat diets for pregnant and postpartum women, malnutrition, or foodborne pathogen infections) may increase the risk of autism and ADHD in infants and young children, as well as mental illnesses in adulthood.

[0003] Studies have shown differences in brain development and cognitive function between breastfed and formula-fed infants. Extensive research also demonstrates that breast milk has a positive effect on infant brain, vision, and cognition development, an effect that can extend from infancy into adolescence. For example, researchers have used MRI to find that breastfed children have more white matter development in the later-maturing frontal lobe; and other studies have found that exclusive breastfeeding increases the cortical thickness of the superior and inferior parietal lobes.

[0004] Human milk oligosaccharides (HMOs), the third largest solid substance in breast milk after lactose and fat, play a vital role in the healthy growth and development of infants, especially in their neural, brain, and cognitive development. Currently, the structures of over 200 HMOs have been identified. Each HMO molecule contains 3-14 monosaccharides linked in a straight or branched chain. Based on their monosaccharide composition, HMOs can be classified into three main categories: fucoidylated human milk oligosaccharides, sialylated human milk oligosaccharides, and neutral human milk oligosaccharides. Significant progress has been made in research on the impact of HMOs on cognitive development over the past two decades. Although clinical trials in human infants present challenges, animal studies consistently demonstrate that HMOs are beneficial for neural development, neurogenesis, neural circuit formation, and memory function.

[0005] However, upon investigation, there are currently very few research reports on the long-term effects of HMOs on individual growth. Summary of the Invention

[0006] The inventors in this case discovered in their research that intervening in human milk oligosaccharides in early life can have a positive effect on cognitive development in adulthood, especially memory ability.

[0007] Specifically, this invention provides the application of human milk oligosaccharides as an active ingredient in the preparation of products that help enhance and / or improve an individual's memory ability in adulthood; The human milk oligosaccharide is selected from one or more of 2'-fucosylated lactose (2'-FL or 2-FL), 3'-sialylated lactose (3'-SL), and 6'-sialylated lactose (6'-SL); The product is an oral product intended for use by minors.

[0008] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharides of the present invention, the ability to enhance and / or improve memory includes assisting in the improvement of memory.

[0009] According to some specific embodiments of the present invention, the application of human milk oligosaccharides of the present invention to enhance and / or improve an individual's memory ability in adulthood includes: Enhance and / or improve an individual's object recognition and memory abilities in adulthood; Enhance and / or improve an individual's spatial memory and pattern separation abilities in adulthood; To enhance and / or improve an individual's learning and memory abilities in adulthood; and / or Increase the number of hippocampal neurons in adults.

[0010] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharides of the present invention, the human milk oligosaccharides are: 2'-Fucose-based lactose; 3'-Sialolactose; 6'-Sialolactose; A combination of 3'-sialyl lactose and 6'-sialyl lactose; or A combination of 2'-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose.

[0011] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharides of the present invention, enhancing and / or improving an individual's adult memory ability includes: enhancing and / or improving an individual's object recognition memory ability in adulthood. The human milk oligosaccharides may be: 2'-fucosylated lactose; 3'-sialylated lactose; 6'-sialylated lactose; a combination of 3'-sialylated lactose and 6'-sialylated lactose; or a combination of 2'-fucosylated lactose, 3'-sialylated lactose, and 6'-sialylated lactose.

[0012] According to some specific embodiments of the present invention, the application of human milk oligosaccharides of the present invention to enhance and / or improve an individual's memory ability in adulthood includes: enhancing and / or improving an individual's spatial memory and pattern separation ability in adulthood. The human milk oligosaccharides may be: 2'-fucosylated lactose; 6'-sialylated lactose; a combination of 3'-sialylated lactose and 6'-sialylated lactose; or a combination of 2'-fucosylated lactose, 3'-sialylated lactose, and 6'-sialylated lactose.

[0013] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, enhancing and / or improving an individual's memory ability in adulthood includes increasing the number of hippocampal neurons in an individual in adulthood. The human milk oligosaccharide is a combination of 2'-fucosylated lactose, 3'-sialylated lactose, and 6'-sialylated lactose.

[0014] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, in the combination of 3'-sialyl lactose and 6'-sialyl lactose, the mass ratio of 3'-sialyl lactose to 6'-sialyl lactose is 1:(1-4), more preferably 1:(1.5-3.5), even more preferably 1:(1.5-2.5) or 1:(2-3).

[0015] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, in the combination of 2'-fucosylated lactose, 3'-sialic acid lactose and 6'-sialic acid lactose, the mass ratio of 2'-fucosylated lactose, 3'-sialic acid lactose and 6'-sialic acid lactose is (1-12):1:(1-4), more preferably (1-12):1:(2-3) or (1-12):1:(1.5-2.5), more preferably (3-11):1:(2-3), and even more preferably (4-10):1:(2-3).

[0016] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, in the combination of 2'-fucosylated lactose, 3'-sialic acid lactose and 6'-sialic acid lactose, the mass ratio of 2'-fucosylated lactose, 3'-sialic acid lactose and 6'-sialic acid lactose is (3-5):1:(2-3), for example (3-5):1:2.

[0017] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, in the combination of 2'-fucosylated lactose, 3'-sialic acid lactose and 6'-sialic acid lactose, the mass ratio of 2'-fucosylated lactose, 3'-sialic acid lactose and 6'-sialic acid lactose is (5-8):1:(2-3), for example (5-7):1:2.

[0018] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, in the combination of 2'-fucosylated lactose, 3'-sialic acid lactose and 6'-sialic acid lactose, the mass ratio of 2'-fucosylated lactose, 3'-sialic acid lactose and 6'-sialic acid lactose is (9-11):1:(2-3), for example (9-11):1:2.

[0019] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharides of the present invention, the product can be a medicine. The term "medicine" as used herein means any substance that provides a beneficial effect to an individual (e.g., a human or animal). The medicine is not necessarily limited to therapeutic drugs requiring market access, but may also include substances that can be used in cosmetics, nutritional products or supplements, health products or supplements, foods (including functional foods), etc. According to some specific embodiments of the present invention, the beneficial effect provided by the product to an individual is that, compared to similar individuals who have not used the human milk oligosaccharides of the present invention or products containing said human milk oligosaccharides, use of the human milk oligosaccharides of the present invention or products containing said human milk oligosaccharides helps to enhance and / or improve an individual's adult memory abilities.

[0020] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharides of the present invention, the product can specifically be a snack, nutritional supplement, dietary supplement, health food, or special medical purpose formula food. The present invention does not impose special requirements on the specific form of the product; for example, it can be milk tablets, candy, or in the form of oral liquid, powder, capsule, etc.

[0021] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, the product is a dairy product.

[0022] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharides of the present invention, the product is infant formula, toddler formula, milk powder, milk tablets, cheese, liquid dairy products, or complementary foods. Since the technical solution of the present invention involves intervention during an individual's adolescence, the product of the present invention is preferably an oral product suitable for infants, children, or adolescents.

[0023] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, the amount of human milk oligosaccharide contained in the product is 10-6000 mg / 100g dry matter.

[0024] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, the amount of human milk oligosaccharide contained in the product is 10-2387 mg / 100g dry matter.

[0025] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, the amount of human milk oligosaccharide contained in the product is 10-1900 mg / 100g dry matter.

[0026] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, the amount of human milk oligosaccharide contained in the product is 20-1800 mg / 100g dry matter.

[0027] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, the amount of human milk oligosaccharide contained in the product is 50-1700 mg / 100g dry matter.

[0028] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, the amount of human milk oligosaccharide contained in the product is 100-1600 mg / 100g dry matter.

[0029] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, the amount of human milk oligosaccharide contained in the product is 200-1500 mg / 100g dry matter.

[0030] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, the amount of human milk oligosaccharide contained in the product is 400-1200 mg / 100g dry matter.

[0031] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, when the human milk oligosaccharide contains 3'-sialic acid lactose, the amount of 3'-sialic acid lactose in the product is usually 10-400 mg / 100g dry matter.

[0032] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, when the human milk oligosaccharide contains 6'-sialyl lactose, the amount of 6'-sialyl lactose in the product is generally 5-800 mg / 100g dry matter. For example, the amount of 6'-sialyl lactose in the product can be 5-50 mg / 100g dry matter, 50-100 mg / 100g dry matter, 100-200 mg / 100g dry matter, 200-500 mg / 100g dry matter, or 500-800 mg / 100g dry matter.

[0033] Understandably, the content of each human milk oligosaccharide in the final product should be appropriately adjusted within the range allowed by relevant standards and regulations for different types of products.

[0034] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharides of the present invention, the individual is a human or animal. In some specific embodiments of the present invention, the individual is a minor with normal memory. In some specific embodiments of the present invention, the individual is a minor with poor memory or memory impairment. The human milk oligosaccharides of the present invention are suitable for assisting in enhancing and / or improving the adult memory ability of the minor.

[0035] According to some specific embodiments of the present invention, in the application of the human milk oligosaccharide of the present invention, the amount of the human milk oligosaccharide given to human individuals is generally 10-200 mg / kg body weight, preferably 40-100 mg / kg body weight.

[0036] According to a specific embodiment of the present invention, the human milk oligosaccharides of the present invention can be derived from commercially available human milk oligosaccharide raw materials with high purity. Unless otherwise specified, the amount or ratio of the human milk oligosaccharides 2'-fucosylated lactose, 3'-sialic acid lactose and 6'-sialic acid lactose in the composition of the present invention is based on the content of human milk oligosaccharides 2'-fucosylated lactose, 3'-sialic acid lactose and 6'-sialic acid lactose in the raw materials.

[0037] The present invention also provides a human milk oligosaccharide composition that helps to enhance and / or improve an individual's memory ability in adulthood, the composition comprising 2'-fucosylated lactose (2'-FL), 3'-sialylated lactose (3'-SL), and 6'-sialylated lactose (6'-SL), wherein the mass ratio of 2'-fucosylated lactose, 3'-sialylated lactose, and 6'-sialylated lactose is (1-12):1:(1-4). According to some specific embodiments of the present invention, the human milk oligosaccharide composition of the present invention that helps to enhance and / or improve an individual's memory ability in adulthood is composed of 2'-fucosylated lactose (2'-FL), 3'-sialylated lactose (3'-SL), and 6'-sialylated lactose (6'-SL).

[0038] According to some specific embodiments of the present invention, in the human milk oligosaccharide composition of the present invention which helps to enhance and / or improve the memory ability of an individual in adulthood, the mass ratio of 2'-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose is (1-12):1:(2-3) or (1-12):1:(1.5-2.5).

[0039] According to some specific embodiments of the present invention, in the human milk oligosaccharide composition of the present invention which helps to enhance and / or improve an individual's memory ability in adulthood, the mass ratio of 2'-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose is (4-10):1:(2-3).

[0040] This invention also provides a method for enhancing and / or improving adult memory in minors, the method comprising: orally administering an effective amount of human milk oligosaccharide to a minor in need. The minor in need may have normal memory, or memory impairment or memory disorders. Administering an effective amount of the human milk oligosaccharide of this invention or a product containing the human milk oligosaccharide helps enhance and / or improve an individual's adult memory by comparison with similar individuals who have not received an effective amount of the human milk oligosaccharide of this invention or a product containing the human milk oligosaccharide. According to some specific embodiments of the invention, the effective amount of the human milk oligosaccharide administered to a human individual is typically 10-200 mg / kg body weight, preferably 40-100 mg / kg body weight.

[0041] The human milk oligosaccharide of this invention, administered orally during an individual's early childhood (e.g., childhood), can enhance and / or improve memory in adulthood. For human individuals, early childhood can refer to infants, children, or adolescents. This invention preferably involves human milk oligosaccharide intervention during infancy or childhood. Adulthood refers to adults, such as young adults, middle-aged individuals, or elderly individuals. This invention preferably enhances and / or improves memory in young adults or middle-aged individuals.

[0042] In summary, this invention explores the effects of early life intervention with human milk oligosaccharides on adult cognitive development. Early life intervention with human milk oligosaccharides has a positive promoting effect on adult cognitive development, especially memory ability, and significantly improves the individual's ability to recognize new objects and new locations in adulthood. Attached Figure Description

[0043] Figure 1 The results show the new object recognition memory outcomes in rats supplemented with 2'-FL during juvenile years.

[0044] Figure 2 The results of novel location recognition memory in rats supplemented with 2'-FL during juvenile stage (easy condition).

[0045] Figure 3 The results show the new object recognition memory outcomes in rats supplemented with 3'-SL and / or 6'-SL during juvenile stage.

[0046] Figure 4 The results of novel location recognition memory in rats supplemented with 3'-SL and / or 6'-SL during juvenile period (easy condition).

[0047] Figure 5 The results of novel location recognition memory (difficult mode) in rats supplemented with 3'-SL and / or 6'-SL during juvenile period are shown.

[0048] Figure 6This shows the results of novel object recognition and memory in rats supplemented with HMO monomers and compositions during juvenile stage.

[0049] Figure 7 This shows the results of novel location recognition memory in rats supplemented with HMO during juvenile stage (easy mode).

[0050] Figure 8 This shows the results of novel location recognition memory (difficult mode) in rats supplemented with HMO during juvenile period.

[0051] Figure 9 The results of staining show the number of newly generated neurons in the hippocampus of rats supplemented with HMO in their early years. Detailed Implementation

[0052] Before further describing specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terminology used in the embodiments of the present invention is for describing specific embodiments and not for limiting the scope of protection of the present invention.

[0053] When numerical ranges are given in the embodiments, it should be understood that, unless otherwise stated in the present invention, the two endpoints of each numerical range and any value between the two endpoints may be selected.

[0054] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art.

[0055] As used herein, the term "enhanced memory" means an improvement in an individual's memory capacity (including short-term memory, long-term memory, memory recall, and / or memory recognition, etc.) compared to the same response in the absence of the human milk oligosaccharides of this invention. This includes, but is not limited to, enhanced object recognition memory, enhanced spatial and pattern separation memory, enhanced learning memory, and increased hippocampal neuron count. In relation to any beneficial effects on memory capacity, the term "enhanced" is synonymous with "strengthened," "improved," or "promoted." In some embodiments of the present invention, particularly in cases where an individual's memory capacity is impaired, declining, or threatened, the term "improving memory capacity" refers to actions that reduce, mitigate, or alleviate the degree of memory impairment or decline, or reduce the risk of memory impairment or decline, including but not limited to: alleviating adverse symptoms associated with memory impairment or decline; slowing or preventing the onset of symptoms before they occur; slowing or halting the progression of memory impairment or decline; slowing or halting the worsening of memory impairment or decline; slowing or halting irreversible damage in the progressive (or chronic) phase of memory impairment or decline; delaying the onset of (progressive) memory impairment or decline; reducing the severity of memory impairment or decline; curing memory impairment or decline; and preventing the occurrence of memory impairment or decline. As used herein, the term "memory impairment" refers to impaired, diminished, or declining learning, memory, and / or cognitive functions, to the point that normal functioning is difficult to maintain or achieve without intervention. Some common signs of memory decline include confusion, short-term or long-term memory loss, and decreased object recognition and / or spatial judgment.

[0056] As used herein, the term "individual" means any animal or human who may benefit from the products of this invention that help improve learning, memory, and / or cognitive function. Generally, when an individual is a human, it can be an infant, child, adolescent, or adult. The term "infant" refers to a human aged 0-3 years from birth; the term "child" refers to a human aged 3-12 years from birth; the term "adolescent" refers to a human aged 12-18 years from birth; the term "adult" refers to a human aged 18 years or older from birth; the term "youth" refers to a human aged 18-39 years from birth; the term "middle-aged" refers to a human aged 40-59 years from birth; and the term "elderly" refers to a human aged 60 years or older from birth. When an individual is an animal, it includes, but is not limited to, birds, bovines, canines, equines, felines, goats, wolves, rodents, sheep, and pigs, preferably mammals.

[0057] In addition to the specific methods, equipment, and materials used in the embodiments, based on the knowledge of those skilled in the art and the description of this invention, any prior art methods, equipment, and materials similar to or equivalent to those described, equipment, and materials in the embodiments of this invention can be used to implement this invention.

[0058] Unless otherwise stated, the experimental methods, detection methods and preparation methods disclosed in this invention all adopt conventional techniques in this technical field.

[0059] In all embodiments and experiments, all human milk oligosaccharides used were commercially available food-grade raw materials. Unless otherwise specified, the amount of human milk oligosaccharides mentioned in this invention is based on the effective content.

[0060] Experimental protocol for early intervention with human milk oligosaccharides to improve adult memory

[0061] 1. Animal models

[0062] The Long Evans rat breeding pairs used in this experiment were purchased from Envigo, USA, for internal breeding of the rats required for the experiment. To ensure that the mother rats could provide adequate care for each pup, litters with more than 10 pups were artificially adjusted to fewer than 10 pups within the first 0 to 1 day after birth. The pups were weaned on day 21 after birth. The rats were housed in an environment with reversed diurnal cycles, i.e., 12 hours of darkness and 12 hours of light (lights off at 9:00 AM and on at 9:00 PM), to adapt to the rats' active cycles.

[0063] 2. HMO intervention

[0064] This study included three different human lactose oligosaccharides (HMOs): 2'-fucosylated lactose (2'-FL), 3'-sialyl lactose (3'-SL), and 6'-sialyl lactose (6'-SL), alone or in combinations of two or three. These HMOs were administered as individual HMOs or in different combinations to juvenile rats (from day 8 to day 35 after birth), forming different group settings with 27 rats in each group. The control group received no HMOs but was given plain drinking water in the same manner as other animals receiving HMOs. The HMOs, either individual or combined, were administered orally by gavage at a daily dose of 400 mg / kg body weight.

[0065] 3. Behavioral research

[0066] After the intervention period ended, all rats in each group were housed in a uniform environment with reversed day and night cycles, fed standard rat diet (without HMOs), and allowed free access to food.

[0067] Rats were assessed behaviorally using memory tests during weeks 32-34, as detailed below: The Novel Object Recognition Test (NORT) is a behavioral task used to assess recognition memory. It measures the time an animal takes to explore a newly introduced object after becoming familiar with a pair of identical objects. In this task, two identical objects are initially presented in a test box, and in the retest phase, one of the objects is replaced with a new object. The time an animal spends exploring the replaced object reflects its object recognition memory ability. NORT is based on an animal's preference for exploring new objects. In the experiment, animals typically spend more time exploring new objects than familiar ones. This preference reflects the animal's ability to recognize and remember new objects.

[0068] In addition to novel object recognition, this experimental study also included a novel object location task (NOLT) to measure the animals' learning and memory abilities. The NOLT utilizes the rats' natural curiosity: when an animal recognizes an object has been moved, it is more inclined to explore the moved object. The variation in the time taken to explore the object at different possible locations was used as a measure of pattern separation. NOLT is a behavioral task used to measure spatial memory and pattern separation (van Goethem, van Hagen & Prickaerts, 2018). The time an animal takes to explore a moved object reflects its spatial memory and pattern separation abilities. The object locations were labeled P1 (location 1), P3 (location 3), and P5 (location 5). Larger numbers indicate greater displacement of the object from the center of the test box, with P1 being the intermediate location (the same as the object location in the acquisition phase). The NOLT study used in this project included two experimental conditions: 1) Easy condition (location 5, P5): In the easy condition, the object was displaced a greater distance from its original location during the test phase. This is considered a low-mode separation condition because displacement of an object over a larger distance is considered easier for the animal to detect; 2) Difficult condition (position 3, P3): Under this condition, the object is displaced a small distance from its original position during the testing phase. This is considered a high-mode separation condition because small-distance displacements are considered harder for the animal to detect.

[0069] 4. Fluorescent staining experiment

[0070] After behavioral studies confirmed that the HMO composition had activity promoting adult cognitive development, the effects of the HMO composition on the rat brain were further investigated. The number of newly generated neurons in the hippocampus was determined using a 5-bromodeoxyuridine staining assay. 5-bromodeoxyuridine is an analog of thymine nucleoside, which can be incorporated into newly synthesized DNA during the DNA synthesis phase of the cell cycle and is commonly used to detect proliferating cells. Female rats in the HMO intervention group underwent the staining experiment 3 days before sacrifice. Rats received intraperitoneal (IP) injections of 5-bromodeoxyuridine at the same time for 3 consecutive days at a dose of 100 mg / kg body weight to ensure consistent labeling of newly generated neurons. Brain samples were collected after rat sacrifice, and hippocampal sections were stained. 5-bromodeoxyuridine-positive cells were manually counted using Cell-Counter in ImageJ.

[0071] 5. Statistical Analysis

[0072] The study used IBM SPSS software version 20.0 for data analysis. A difference was considered statistically significant when the reported p-value was less than 0.05, and statistical trend was defined as a p-value less than or equal to 0.1.

[0073] Example 1: Early intervention with 2'-FL improves adult memory

[0074] In this embodiment, following the aforementioned experimental protocol for "Early Intervention of Test Substances to Improve Adult Memory," the effects of early intervention with 2'-fucosylated lactose (2'-FL) on the learning and memory abilities of adult rats were tested. 2'-FL was administered as a single HMO to juvenile rats (from day 8 to day 35 after birth), resulting in the following group setup: a control group (receiving drinking water) and a 2'-FL intervention group. The control group did not contain any HMOs but was given ordinary drinking water in the same manner as other animals receiving HMOs. Each group consisted of 27 rats. 2'-FL was administered orally by gavage at a dose of 400 mg / kg body weight daily.

[0075] Figure 1 This study shows the results of novel object recognition memory in rats supplemented with 2'-FL during juvenile stage. The NORT experiment was conducted at weeks 32-34 of rat life, and data are expressed as the mean and standard error of the discrimination index. The results showed that the novel object discrimination index was increased in the 2'-FL intervention group compared to the control group. This indicates that 2'-FL intervention during juvenile stage has a significant effect on improving memory abilities in adulthood.

[0076] Figure 2This study shows the results of novel location recognition and memory in rats supplemented with 2'-FL during juvenile stage (easy mode). The NOLT experiment was conducted at weeks 32-34 of rat life, and data are expressed as the mean and standard error of the number of times the experimental animals correctly identified and explored objects in novel locations during the test phase. The results showed that the 2'-FL intervention group had a higher number of correct answers compared to the control group. This indicates that 2'-FL intervention during juvenile stage has a significant effect on improving learning and memory abilities in adulthood.

[0077] Example 2: Early intervention with 3'-SL and / or 6'-SL improves adult memory

[0078] In this embodiment, following the aforementioned "Experimental Protocol for Early Intervention of Test Substances to Improve Adult Memory," the effects of early intervention with 3'-sialyl lactose (3'-SL) and / or 6'-sialyl lactose (6'-SL) on learning and memory abilities in adult rats were tested. These HMOs were administered as single HMOs or in combination during juvenile rats (from day 8 to day 35 after birth), forming the following groups: control group (receiving drinking water), HMO single group, and HMO combination group (3'-SL + 6'-SL, with a mass ratio of 3'-SL to 6'-SL of 1:2). The control group received no HMOs but was given ordinary drinking water in the same manner as other animals receiving HMOs; each group consisted of 27 rats. The HMO single group or combination group was administered 400 mg / kg body weight orally by gavage daily (i.e., the HMO combination group was administered 133 mg / kg body weight of 3'-SL + 267 mg / kg body weight of 6'-SL orally by gavage daily).

[0079] Figure 3 This study demonstrates the results of novel object recognition and memory in rats supplemented with 3'-SL and / or 6'-SL during juvenile stage. The NORT experiment was conducted at weeks 32-34 of rat life, and data are expressed as mean and standard error of the discrimination index. Results showed that the novel object discrimination index was increased in the sialic acid intervention group compared to the control group. In particular, the 3'-SL+6'-SL combination group had the highest object recognition index, indicating that sialic acid intervention during juvenile stage significantly improves memory ability in adulthood, and that the two sialic acid oligosaccharides have a synergistic effect.

[0080] Figure 4The results show the novel location recognition memory outcomes (easy condition) of rats supplemented with 3'-SL and / or 6'-SL during juvenile stage. The NOLT experiment was conducted at weeks 32-34 of rat life, and data are expressed as the mean and standard error of the number of correct answers. The results showed that, compared with the control group, the 6'-SL group and the 3'-SL+6'-SL intervention group had increased novel location discrimination indices. This indicates that sialic acid oligosaccharide intervention during juvenile stage, especially 6'-SL and the combination of 3'-SL and 6'-SL, has a significant effect on improving learning and memory abilities in adulthood.

[0081] Figure 5 The results show the novel location recognition and memory outcomes (difficult mode) of rats supplemented with 3'-SL and / or 6'-SL during juvenile stage. The closer the novel location is to the center, the greater the recognition difficulty. Therefore, the performance of rats in the difficult mode can indicate their performance in handling complex tasks. The experimental results showed that, compared with the control group, the novel location discrimination index was increased in the 6'-SL group and the 3'-SL+6'-SL intervention group. This indicates that sialic acid oligosaccharide intervention during juvenile stage, especially 6'-SL and the combination of 3'-SL and 6'-SL, has a significant effect on improving learning and memory abilities in adulthood.

[0082] Example 3: Early intervention with HMO improves adult memory

[0083] In this embodiment, following the aforementioned "Experimental Protocol for Early Intervention of Test Substances to Improve Adult Memory," three different human lactose oligosaccharides (HMOs) were tested: 2'-fucosylated lactose (2'-FL), 3'-sialylated lactose (3'-SL), and 6'-sialylated lactose (6'-SL). These HMOs were administered as individual HMOs or in different combinations to juvenile rats (from day 8 to day 35 after birth), resulting in the following group settings: control group (receiving drinking water), HMO monomer groups (2'-FL, 3'-SL, and 6'-SL groups), HMO composition 1, HMO composition 2, and HMO composition 3. Each control and HMO monomer group consisted of 27 rats, and each HMO combination group consisted of 27 rats. The control group received no HMOs but was given ordinary drinking water in the same manner as other animals receiving HMOs. The daily dose of either the HMO monomer or the combination administered orally via gavage was 400 mg / kg body weight (the gavage dose for the HMO combination group was the total HMO dose of 400 mg / kg body weight).

[0084] The experimental group information is shown in Table 1.

[0085] Table 1. Experimental group settings

[0086] Figure 6This study shows the results of novel object recognition and memory in rats supplemented with HMOs during juvenile stage. The NORT experiment was conducted at 32-34 weeks of rat life, and data are expressed as the mean and standard error of the discrimination index. The results showed that the novel object discrimination index was increased in the HMO combination intervention groups compared to the control group. In particular, HMO combination 1 group had the highest object recognition index, followed by HMO combination 3 group and the 6'-SL group, indicating that intervention with human milk oligosaccharides during juvenile stage significantly improves memory ability in adulthood, and the three HMO combinations showed a certain synergistic effect.

[0087] Figure 7 The results show the novel location recognition memory outcomes (easy mode) of rats supplemented with HMOs during juvenile stage. The NOLT experiment was conducted at 32-34 weeks of age, and data are expressed as the mean and standard error of the number of correct answers. The results showed that the novel location discrimination index was increased in HMO combination 1 and HMO combination 3 groups compared with the control group. This indicates that intervention with human milk oligosaccharides during juvenile stage, especially the combination of 2'-FL, 3'-SL, and 6'-SL, has a significant effect on improving learning and memory abilities in adulthood.

[0088] Figure 8 The results show the novel location recognition and memory outcomes (difficult mode) of rats supplemented with HMOs during juvenile stage. The closer the novel location is to the center, the greater the recognition difficulty. Therefore, the performance of rats in the difficult mode can indicate their performance in handling complex tasks. In this experiment, no meaningful statistical data were obtained for the 2'-FL group and the HMO composition 3 group due to operational abnormalities, and they are not shown in the figure. The experimental results show that, compared with the control group, the novel location discrimination index of the HMO composition 1 group and the HMO composition 2 group intervention group was increased. This indicates that HMO composition intervention during juvenile stage, especially the combination of 2'-FL, 3'-SL, and 6'-SL, has a significant effect on improving learning and memory abilities in adulthood, especially the ability to handle complex situations.

[0089] After behavioral experiments confirmed the positive impact of early intervention with HMO combinations on cognitive development in adulthood, the number of newly generated neurons in the brains of rats in the HMO combination groups was measured. Fluorescent staining was used to analyze the differences in newly generated neurons in the brains of rats in the control group and the HMO combination groups 1, 2, and 3. Figure 9 The results show the staining results of the number of newly generated neurons in the hippocampus of rats supplemented with HMOs during childhood. The results indicate that, compared with the control group, the intervention groups (HMO composition 1 and HMO composition 3) had an increased number of fluorescently positive cells, indicating an increase in the number of newly generated neurons in the hippocampus. This suggests that intervention with HMO compositions during childhood, especially the combinations of 2'-FL, 3'-SL, and 6'-SL, may enhance cognitive development in adulthood by increasing the number of newly generated neurons in the hippocampus.

[0090] The above description of the embodiments is provided to enable those skilled in the art to understand and use the invention. It will be apparent to those skilled in the art that various modifications can be made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present invention is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the invention should be within the protection scope of the present invention.

Claims

1. Application of human milk oligosaccharides as an active ingredient in the preparation of products that help enhance and / or improve an individual's memory abilities in adulthood; in, The human milk oligosaccharide is selected from one or more of 2'-fucosylated lactose (2'-FL), 3'-sialylated lactose (3'-SL), and 6'-sialylated lactose (6'-SL); The product is an oral product intended for use by minors.

2. The application according to claim 1, wherein, Enhancing and / or improving an individual's memory abilities in adulthood includes: Enhance and / or improve an individual's object recognition and memory abilities in adulthood; Enhance and / or improve an individual's spatial memory and pattern separation abilities in adulthood; To enhance and / or improve an individual's learning and memory abilities in adulthood; and / or Increase the number of hippocampal neurons in adults.

3. The application according to claim 1, wherein, Human milk oligosaccharides are: 2'-Fucose-based lactose; 3'-Sialolactose; 6'-Sialolactose; A combination of 3'-sialyl lactose and 6'-sialyl lactose; or A combination of 2'-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose.

4. The application according to claim 1 or 3, wherein, Enhancing and / or improving an individual's memory abilities in adulthood includes: enhancing and / or improving an individual's object recognition memory abilities in adulthood.

5. The application according to claim 1, wherein, Enhancing and / or improving an individual's memory abilities in adulthood includes: enhancing and / or improving an individual's spatial memory and pattern discrimination abilities in adulthood; The human milk oligosaccharide is: 2'-Fucose-based lactose; 6'-Sialolactose; A combination of 3'-sialyl lactose and 6'-sialyl lactose; or A combination of 2'-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose.

6. The application according to claim 1, wherein, Enhancing and / or improving an individual's memory capacity in adulthood includes increasing the number of hippocampal neurons in an individual's adulthood; The human milk oligosaccharide is a combination of 2'-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose.

7. The application according to claim 3 or 5, wherein, In the combination of 3'-sialyl lactose and 6'-sialyl lactose, the mass ratio of 3'-sialyl lactose to 6'-sialyl lactose is 1:(1-4). Optionally, in the combination of 2'-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose, the mass ratio of 2'-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose is (1-12):1:(1-4).

8. The application according to claim 1, wherein, The products are snacks, nutritional supplements, dietary supplements, health foods, or foods for special medical purposes.

9. The application according to claim 8, wherein, The product contains 10-6000 mg / 100g of human milk oligosaccharides.

10. A human milk oligosaccharide composition that helps to enhance and / or improve an individual's memory ability in adulthood, said composition comprising 2'-fucosylated lactose (2'-FL), 3'-sialylated lactose (3'-SL) and 6'-sialylated lactose (6'-SL), wherein the mass ratio of 2'-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose is (1-12):1:(1-4).

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