Nutritional composition, food and medicine comprising the nutritional composition, and uses thereof

Abstract

The present invention relates to a nutritional composition, a food and a medicine comprising the nutritional composition, and their uses. The nutritional composition comprises medium and long chain fatty acid triglycerides and human milk oligosaccharides, and the weight ratio of the medium and long chain fatty acid triglycerides to the human milk oligosaccharides is 1:1 to 9:1. The nutritional composition of the present invention can synergistically promote bone development, particularly improve bone density.

Description

Technical Field

[0001] The present invention relates to the field of food, and particularly to a nutritional composition, foods and drugs comprising the nutritional composition, and their uses. Background Art

[0002] Bone is a highly active connective tissue that can repair its own microdamage through bone metabolism and bone remodeling to maintain the homeostatic balance of bone structure, load, and calcium content. At the same time, bone also acts as an endocrine organ to regulate metabolic processes and is an important organ for maintaining human life. Since the neonatal period, bone mass increases with age until bone development matures in adolescence and bone mass reaches its peak. Studies have found that paying attention to increasing bone mass during adolescence can achieve an individual peak bone mass in young adulthood. Therefore, the growth and development of bone during adolescence is crucial for bone maturity. Some studies have pointed out that bone mineral density (BMD) during adolescence determines the peak bone mass in adulthood and the subsequent bone loss rate. A decrease in BMD can lead to insufficient bone mass, thereby triggering a series of bone diseases such as rickets and osteomalacia, seriously affecting the bone health of adolescents. During this period, nutritional supplementation is an important factor affecting peak bone mass. Intaking nutrients can increase peak bone mass, thereby promoting bone formation. Therefore, seeking a natural functional nutrient to increase adolescent BMD and enhance bone formation is of great significance for increasing bone mass, preventing bone diseases such as adolescent fractures and senile osteoporosis, and maintaining bone health.

[0003] Bone contains an extracellular protein matrix (osteoid), in which osteocytes are scattered and a mineral component composed of calcium salts and other minerals is located within the extracellular matrix. Bone undergoes remodeling during the resorption process, in which case bone is degraded by osteoclasts and then replaced (reformed) by osteoblasts. Remodeling occurs to regulate calcium homeostasis, repair bone damaged by daily stress, and shape bone during growth or changes in mechanical stress patterns. Osteoclasts degrade bone in a specific area and then undergo apoptosis. Osteoblasts rebuild new bone and mediate its remineralization. During remineralization, some osteoblasts are encapsulated within calcified material and then become osteocytes.

[0004] Once peak bone content has been reached from infancy to early adulthood, the resorption process is almost completely accompanied by reformation. However, as people age, the effective regulation of the remodeling system weakens. With the onset of female menopause, these two processes may become asynchronous, with resorption predominating. In addition, with aging, there is bone mineral density loss, bone microstructure damage, and other changes, all of which lead to an increased risk of fracture. The process of bone loss is often gradual, with no obvious symptoms presented before the proper progression of the disease. Therefore, it is important to maintain bone quality and density over time so that bone does not become fragile with aging.

[0005] Therefore, there is a need to provide a nutritional composition that can promote bone development, especially for infants and children. Summary of the Invention

[0006] The present invention is made in view of the above problems existing in the prior art.

[0007] In a first aspect, the present invention relates to a nutritional composition comprising:

[0008] medium and long-chain fatty acid triglycerides; and

[0009] human milk oligosaccharides,

[0010] wherein the weight ratio of the medium and long-chain fatty acid triglycerides to the human milk oligosaccharides is from 1:1 to 9:1.

[0011] The inventors unexpectedly found in their research that when medium and long-chain fatty acid triglycerides and human milk oligosaccharides are used in combination in a certain weight ratio, they can exert a synergistic effect and synergistically promote bone development, particularly increase bone density.

[0012] In a second aspect, the present invention relates to a food comprising the nutritional composition according to the first aspect.

[0013] In a third aspect, the present invention relates to a medicament comprising the nutritional composition according to the first aspect.

[0014] In a fourth aspect, the present invention relates to the use of the nutritional composition according to the first aspect or the food according to the second aspect for non-therapeutic purposes of promoting bone development, particularly increasing bone density.

[0015] In a fifth aspect, the present invention relates to the use of the nutritional composition according to the first aspect in the preparation of a medicament for promoting bone development, particularly increasing bone density. Brief Description of the Drawings

[0016] To more clearly illustrate the technical solutions of the present invention, the drawings required for describing the embodiments will be briefly described below. It should be understood that these drawings are only for the convenience of those skilled in the art to more easily understand the present invention, and are not intended to limit the scope of the present invention.

[0017] Figure 1 Typical diagram showing the fluorescence intensity of zebrafish skull after sample treatment.

[0018] Figure 2 Bar chart showing the fluorescence intensity of zebrafish skull after sample treatment. Compared with the normal control group: * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001. Detailed Embodiments

[0019] To make the inventive purpose, technical solution and beneficial technical effects of this application clearer, the following will provide a detailed description of this application. It should be noted that the various aspects, features, embodiments, and their advantages described in this application can be compatible and / or combined together.

[0020] Unless otherwise specified, the meanings of technical terms or scientific and technical terms in this specification are the same as those generally understood by those skilled in the art.

[0021] In this application, unless otherwise specified, the temperature is room temperature (25°C), the atmosphere is air, and the pressure is atmospheric pressure.

[0022] In this application, the term "medium-chain fatty acid" refers to fatty acids having 6-12 carbon atoms in the carbon chain, such as caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecanoic acid, dodecanoic acid, etc.; the term "long-chain fatty acid" refers to fatty acids having more than 14 carbon atoms, generally 14-30 carbon atoms, such as myristic acid, palmitic acid, oleic acid, linoleic acid, stearic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid, etc. The terms "medium-chain triglyceride" and "long-chain triglyceride" respectively refer to the esterification reaction products of "medium-chain fatty acid" and "long-chain fatty acid" with glycerol; the term "medium- and long-chain triglyceride" refers to triglycerides that contain both medium-chain fatty acid residues and long-chain fatty acid residues in the molecular structure.

[0023] In this application, the term "human milk oligosaccharide", also known as "human milk oligosaccharide", is a class of complex oligosaccharides formed by glycosidic bonds connecting structural units such as monosaccharides and their derivatives, sialic acid, etc.

[0024] In this application, the term "non-therapeutic purpose" has its conventional meaning in the art, which includes, for example, nutritional and / or health care purposes, which may be, for example, for the preparation of foods such as ordinary foods, functional foods, special foods (including, for example, infant formula foods, foods for special medical purposes, and health foods), etc.

[0025] The present invention relates to nutritional compositions, foods and drugs including such nutritional compositions, and their uses.

[0026] The following will provide a detailed description of the present invention.

[0027] Nutritional Composition

[0028] In a first aspect, the present invention relates to a nutritional composition comprising medium-chain triglycerides and human milk oligosaccharides, wherein the weight ratio of the medium-chain triglycerides to the human milk oligosaccharides is from 1:1 to 9:1. As an example, the mass ratio of the medium-chain triglycerides to the human milk oligosaccharides can be 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or a range defined by any two of them.

[0029] The inventors unexpectedly found in their research that medium-chain triglycerides and human milk oligosaccharides (especially 2'-fucosyllactose, 3'-sialyllactose and / or lacto-N-neotetraose) can synergistically promote bone development when used in combination in a specific ratio. The bone development can, for example, include increasing bone density.

[0030] In some embodiments, the mass ratio of the medium-chain triglycerides to the human milk oligosaccharides can be from 2:1 to 9:1; within this range, the nutritional composition can more significantly synergistically promote bone development, such as increasing bone density.

[0031] The present invention has no particular requirements for the human milk oligosaccharides used, and the human milk oligosaccharides commonly used in the art can be used. The human milk oligosaccharides can be used in pure form or in a non-pure form rich in human milk oligosaccharides.

[0032] Human milk oligosaccharides (also known as milk oligosaccharides or human milk oligosaccharides, Human Milk Oligosaccharides) are the third largest solid component in breast milk, second only to lactose and fat, and have important biological activities. Classified according to representative groups, human milk oligosaccharides can be divided into fucosylated human milk oligosaccharides (such as 2'-fucosyllactose), sialylated human milk oligosaccharides (such as 3'-sialyllactose) and non-fucosylated non-sialylated human milk oligosaccharides (such as lacto-N-neotetraose).

[0033] In some embodiments, the human milk oligosaccharide is selected from one or more of the following: fucosylated human milk oligosaccharides, sialylated human milk oligosaccharides, non-fucosylated and non-sialylated human milk oligosaccharides. For example, the human milk oligosaccharide can be: one or more fucosylated human milk oligosaccharides; one or more sialylated human milk oligosaccharides; one or more non-fucosylated and non-sialylated human milk oligosaccharides; one or more fucosylated human milk oligosaccharides and one or more sialylated human milk oligosaccharides; one or more fucosylated human milk oligosaccharides and one or more non-fucosylated and non-sialylated human milk oligosaccharides; one or more sialylated human milk oligosaccharides and one or more non-fucosylated and non-sialylated human milk oligosaccharides; one or more fucosylated human milk oligosaccharides, one or more sialylated human milk oligosaccharides, and one or more non-fucosylated and non-sialylated human milk oligosaccharides.

[0034] In some embodiments, the fucosylated human milk oligosaccharide is, for example, selected from one or more of the following: 2'-fucosyllactose (2'-FL), 3-fucosyllactose, lacto-N-fucopentaose I, lacto-N-difucohexaose I, lacto-N-difucohexaose II, lacto-bifucosyltetraose.

[0035] In some embodiments, the sialylated human milk oligosaccharide is, for example, selected from one or more of the following: 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL).

[0036] In some embodiments, the non-fucosylated and non-sialylated human milk oligosaccharide is, for example, selected from one or more of the following: lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), lacto-N-triose II, and lacto-N-diol.

[0037] In some embodiments, the human milk oligosaccharide is, for example, selected from one of the following: 2'-fucosyllactose; 3'-sialyllactose; lacto-N-neotetraose; 2'-fucosyllactose and 3'-sialyllactose; 2'-fucosyllactose and lacto-N-neotetraose; 3'-sialyllactose and lacto-N-neotetraose; 2'-fucosyllactose, 3'-sialyllactose, and lacto-N-neotetraose.

[0038] In some embodiments, the human milk oligosaccharide is, for example, selected from one of the following: 2'-fucosyllactose; lacto-N-neotetraose; 2'-fucosyllactose and lacto-N-neotetraose.

[0039] For example, in some preferred embodiments, the human milk oligosaccharides are 2'-fucosyllactose, 3'-sialyllactose, and lacto-N-neotetraose, and the weight ratio of the medium-chain and long-chain fatty acid triglycerides to the human milk oligosaccharides is from 3:1 to 9:1; in some preferred embodiments, the human milk oligosaccharides are any two selected from 2'-fucosyllactose, 3'-sialyllactose, and lacto-N-neotetraose, and the weight ratio of the medium-chain and long-chain fatty acid triglycerides to the human milk oligosaccharides is from 5:1 to 7:1; in some preferred embodiments, the human milk oligosaccharides are 2'-fucosyllactose, and the weight ratio of the medium-chain and long-chain fatty acid triglycerides to the human milk oligosaccharides is from 5:1 to 9:1; within the above preferred ranges, the synergistic effect of the nutritional composition in promoting bone development, particularly in enhancing bone density, is more significant.

[0040] When the human milk oligosaccharides are a mixture of any two or more, the ratio between the components is not particularly limited and can be appropriately selected in consideration of the other effects of each specific human milk oligosaccharide; for example, the mass ratio between any two specific human milk oligosaccharides can be between 20:1 and 1:20, such as 15:1 to 1:15.

[0041] For example, when the human milk oligosaccharides include fucosylated human milk oligosaccharides (such as 2'-FL) and sialylated human milk oligosaccharides (such as 3'-SL), the mass ratio of the fucosylated human milk oligosaccharides / sialylated human milk oligosaccharides, such as the 2'-FL / 3'-SL mass ratio, can be 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, or a range defined by any two of them.

[0042] For example, when the human milk oligosaccharides include fucosylated human milk oligosaccharides (such as 2'-FL) and non-fucosylated and non-sialylated human milk oligosaccharides (such as LNnT), the mass ratio of fucosylated human milk oligosaccharides to non-fucosylated and non-sialylated human milk oligosaccharides, such as the 2'-FL / LNnT mass ratio, can be 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, or a range defined by any two of them.

[0043] For example, when the human milk oligosaccharides include sialylated human milk oligosaccharides (such as 3'-SL) and non-fucosylated and non-sialylated human milk oligosaccharides (such as LNnT), the mass ratio of sialylated human milk oligosaccharides to non-fucosylated and non-sialylated human milk oligosaccharides, such as the 3'-SL / LNnT mass ratio, can be 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, or a range defined by any two of them.

[0044] For example, when the human milk oligosaccharides include fucosylated human milk oligosaccharides (such as 2’-FL), sialylated human milk oligosaccharides (such as 3’-SL), and non-fucosylated and non-sialylated human milk oligosaccharides (such as LNnT), the mass ratio of fucosylated human milk oligosaccharides to sialylated human milk oligosaccharides, such as the 2’-FL / 3’-SL mass ratio, the mass ratio of fucosylated human milk oligosaccharides to non-fucosylated and non-sialylated human milk oligosaccharides, such as the 2’-FL / LNnT mass ratio, and the mass ratio of sialylated human milk oligosaccharides to non-fucosylated and non-sialylated human milk oligosaccharides, such as the 3’-SL / LNnT mass ratio, can each independently be 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, or a range defined by any two of them. For example, in one embodiment, when the human milk oligosaccharides are a combination of fucosylated human milk oligosaccharides, sialylated human milk oligosaccharides, and non-fucosylated and non-sialylated human milk oligosaccharides, based on 4 parts by weight of non-fucosylated and non-sialylated human milk oligosaccharides (such as LNnT), the amount range of fucosylated human milk oligosaccharides (such as 2’-FL) is 8 to 28, preferably 16 to 24 parts by weight, and / or the amount range of sialylated human milk oligosaccharides (such as 3’-SL) is 1 to 8, preferably 1 to 4 parts by weight.

[0045] Medium-chain triglycerides (MLCT) are a type of structural lipid in which triglyceride molecules contain both medium-chain fatty acids and long-chain fatty acids, and they have different physical and chemical properties, metabolic characteristics, and nutritional values from long-chain triglycerides (LCT) or medium-chain triglycerides (MCT). Research (see, for example, Yuan Tinglan, Composition and Metabolic Characteristics of Medium-chain Triglycerides in Human Milk Fat [D], Jiangnan University, 2021) shows that medium-chain triglycerides (MLCT) are not simply equivalent to a physical mixture of long-chain triglycerides (LCT) and medium-chain triglycerides (MCT) (certainly not equivalent to a simple mixture of long-chain fatty acids and medium-chain fatty acids), and the latter do not possess the physical and chemical properties, metabolic characteristics, and nutritional values of the former.

[0046] The present invention has no particular requirements for the medium- and long-chain fatty acid triglycerides used, and the medium- and long-chain fatty acid triglycerides commonly used in the art can be used. The medium- and long-chain fatty acid triglycerides can be used in the form of pure products or in the form of non-pure products rich in medium- and long-chain fatty acid triglycerides. In some embodiments, the medium- and long-chain fatty acid triglycerides of the present invention contain C 6 -C 12 (for example, C 6 、C 7 、C 8 、C 9 、C 10 、C 11 、C 12 、or the range defined by any two of them) fatty acid residues and C 14 -C 30 (for example, C 14 、C 15 、C 16 、C 17 、C 18 、C 19 、C 20 、C 21 、C 22 、C 23 、C 24 、C 25 、C 26 、C 27 、C 28 、C 29 、C 30 、or the range defined by any two of them) fatty acid residues, where:

[0047] The C 6 -C 12 fatty acid residues are derived from one or more of the following: caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecanoic acid, lauric acid, etc., preferably caproic acid, caprylic acid, capric acid, and lauric acid; and / or

[0048] The C 14 -C 30 fatty acid residues are derived from one or more of the following: myristic acid, palmitic acid, margaric acid, oleic acid, linoleic acid, stearic acid, nonadecanoic acid, eicosapentaenoic acid, heneicosanoic acid, docosatetraenoic acid, docosahexaenoic acid, tricosanoic acid, tetracosenoic acid, pentacosapentaenoic acid, docosahexaenoic acid, arachidonic acid, etc., preferably myristic acid, palmitic acid, oleic acid, linoleic acid, stearic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid.

[0049] In some embodiments, in the medium- and long-chain fatty acid triglycerides, the C 6 -C 12The weight ratio of the fatty acid residue to the C 14 -C 30 is from 0.124 to 2.000. As an example, the weight ratio of the C 6 -C 12 fatty acid residue to the C 14 -C 30 fatty acid residue can be 0.124, 0.125, 0.130, 0.140, 0.150, 0.160, 0.170, 0.180, 0.190, 0.200, 0.300, 0.400, 0.500, 0.600, 0.700, 0.800, 0.900, 1.000, 1.100, 1.200, 1.300, 1.400, 1.500, 1.600, 1.700, 1.800, 1.900, 2.000, or within the range defined by any two of them.

[0050] In some embodiments, the nutritional composition of the present invention may consist of medium-chain and long-chain fatty acid esters and human milk oligosaccharides.

[0051] Food

[0052] In a second aspect, the present invention relates to a food comprising the nutritional composition according to the first aspect.

[0053] In some embodiments, the food is an infant food such as infant formula.

[0054] In some embodiments, based on the weight of the food, the content of medium-chain and long-chain triglycerides can be from 0.1 to 30% by weight. As an example, based on the weight of the food, the content of medium-chain and long-chain triglycerides can be 0.1, 0.2, 0.5, 0.8, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30% by weight, or within the range defined by any two of them.

[0055] In some embodiments, based on the weight of the food, the content of human milk oligosaccharides can be from 0.01 to 6% by weight. As an example, based on the weight of the food, the content of human milk oligosaccharides can be 0.01, 0.02, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6% by weight, or within the range defined by any two of them.

[0056] In some embodiments, the food can be, for example, functional foods and special foods (infant formula, foods for special medical purposes, or health foods).

[0057] In some embodiments, the food is a general food or a special food. By way of example, the food may be selected from dairy products, candies, beverages, breads, and biscuits. For example, the food may be selected from milk powder or fermented foods.

[0058] In some embodiments, the milk powder is infant formula milk powder.

[0059] In addition, it is readily understood by those skilled in the art that, in addition to the nutritional composition of the first aspect of the present invention, the food further comprises one or more selected from the following: raw cow milk, demineralized whey powder, concentrated whey protein, lactose, edible vegetable blended oil, fructooligosaccharide, galactooligosaccharide, nucleotides, choline, vitamins, minerals, DHA, and taurine.

[0060] For example, when the food is milk powder, in addition to the nutritional composition, the milk powder may further comprise proteins such as α-lactalbumin and milk fat globule membrane protein; saccharides such as lactose; lipids; minerals such as calcium, iron, phosphorus, etc.; vitamins; and other additives such as whey powder, choline bitartrate, docosahexaenoic acid, arachidonic acid, walnut oil, etc.

[0061] The food may be produced by a preparation method commonly used in the art, which is well known to those skilled in the art and will not be elaborated herein.

[0062] All of the above descriptions regarding the nutritional composition of the first aspect of the present invention apply herein and will not be elaborated herein.

[0063] Medicine

[0064] In a third aspect, the present invention relates to a medicine comprising the nutritional composition according to the first aspect.

[0065] The present invention has no particular requirement for the dosage form of the medicine, and common dosage forms in the art may be adopted. By way of example, the medicine of the present invention may be a solid preparation, such as powder, granule, tablet, capsule, dripping pill, pill, etc.; semi-solid preparation, such as ointment, cream, eye ointment, gel, etc.; liquid preparation, such as oral liquid preparation, such as syrup, mixture, suspension, emulsion, etc.

[0066] It is readily understood by those skilled in the art that, depending on the different dosage forms and application scenarios of the medicine, the medicine may further comprise various pharmaceutically acceptable excipients. In addition, the medicine of the present invention may be prepared by a method commonly used in the art, which is well known to those skilled in the art and will not be elaborated herein.

[0067] All of the above descriptions regarding the nutritional composition of the first aspect of the present invention apply herein and will not be elaborated herein.

[0068] Use

[0069] In a fourth aspect, the present invention relates to the use of the nutritional composition according to the first aspect or the food according to the second aspect for the non-therapeutic purpose of promoting bone development, particularly increasing bone density.

[0070] The promotion of bone development may, for example, include improving (increasing bone density).

[0071] The promotion of bone development may, for example, include, but is not limited to, promoting bone development in infants and / or children and / or adolescents, particularly increasing bone density.

[0072] The non-therapeutic purpose may, for example, include nutritional and / or health care purposes, such as for the preparation of foods such as ordinary foods, functional foods, health foods, or health products, etc.

[0073] In some embodiments, when the nutritional composition or food is orally administered to a human body, the dosage is such that the dosage of the medium and long-chain fatty acid triglycerides is 0.1 to 30 g / day, preferably 1.7 to 30 g / day. As an example, the dosage of the medium and long-chain fatty acid triglycerides can be 0.1, 0.2, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 g / day, or within the range defined by any two of them.

[0074] In some embodiments, when the nutritional composition or food is orally administered to a human body, the dosage is such that the dosage of the human milk oligosaccharides is 0.05 to 25 g / day, preferably 0.1 to 25 g / day. As an example, the dosage of the human milk oligosaccharides can be 0.05, 0.1, 0.2, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 g / day, or within the range defined by any two of them, and the dosage of the medium and long-chain fatty acid triglycerides can be converted according to the ratio to the human milk oligosaccharides.

[0075] In a fifth aspect, the present invention relates to the use of the nutritional composition according to the first aspect in the preparation of a medicament for promoting bone development, particularly increasing bone density.

[0076] All of the above descriptions of the nutritional composition in the first aspect, the food in the second aspect, and the medicament in the third aspect of the present invention are applicable here and will not be repeated.

[0077] Examples

[0078] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention will be further described in detail below in conjunction with embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0079] Experimental principle description

[0080] The embodiment adopts a zebrafish experimental scheme. Compared with traditional animal tests (piglets, mice and rats) and human experiments, the zebrafish experimental scheme has many advantages in the efficacy evaluation experiment of promoting growth (bone) development. (1) Zebrafish has been widely used in recent years as an important model for the establishment of disease models and drug discovery. The research on its bones has a physiological and genetic basis: the bone development of zebrafish is extremely similar to that of other vertebrates. Bones are composed of two different tissues, cartilage and bone. (2) At the cellular level, the osteoblasts and osteoclasts of zebrafish have functions similar to those of mammals. Its osteoclasts have bone resorption effects during the early embryonic development of zebrafish. (3) At the anatomical level, the morphology and development process of the bones of zebrafish larvae and adults have been basically clarified. Its bone development process is highly consistent with that of mammals, that is, bone formation occurs through two mechanisms: intramembranous ossification and endochondral ossification respectively. (4) In addition, zebrafish has a high degree of genetic similarity with humans and is suitable for gene manipulation techniques, such as gene knockout and knock-in. The application of these techniques enables researchers to more precisely study the effects of specific genes on brain development. (5) Zebrafish have strong reproductive capacity, fast development and short sexual maturity cycles, which makes them very suitable for high-throughput experiments. High-throughput experiments can greatly accelerate the research process and improve experimental efficiency. (6) The zebrafish model also has strong translational relevance, that is, its research results are more easily generalized to humans. This makes zebrafish have important application value in the efficacy evaluation of promoting brain development.

[0081] Raw materials

[0082] The sources and specifications of the raw materials used in the following embodiments are as follows:

[0083] Medium and long-chain fatty acid triglycerides: sourced from medium and long-chain fatty acid edible oil (provided by Qingdao Haizhiyuan Life Science and Technology Co., Ltd., batch number Y1501-23070101), purity 69.8%;

[0084] 2'-Fucosyllactose: provided by DSM Vitamins Trading (Shanghai) Co., Ltd., SHANGHAI, purity 99.2%, batch number: 21107002;

[0085] 3'-Sialyllactose: Provided by DSM Vitamins Trading(Shanghai)Co., Ltd., SHANGHAI, with a purity of 95.2%, batch number: DK21294101;

[0086] Lacto-N-neotetraose: Provided by DSM Vitamins Trading(Shanghai)Co., Ltd., DTL DutyPaid., Shanghai, with a purity of 100.5%, batch number: 20454001;

[0087] Swisse “Calcium&Vitamin D”: Swisse “Calcium&Vitamin D”, batch number 33152A, provided by Swisse, specification: each tablet contains 333.33 mg of calcium;

[0088] Wild-type AB strain zebrafish: Reproduced and provided by the fish breeding center of Hangzhou Huante Biotechnology Co., Ltd., with the experimental animal use license number: SYXK(Zhe)2022 - 0004.

[0089] Positive control

[0090] Swisse “Calcium&Vitamin D”

[0091] In the following examples, AB strain zebrafish were used for animal experiments.

[0092] 1. Detection materials

[0093] 1.1. Sample preparation information

[0094] For MLCT, 2'-FL, LNnT, 3'-SL, the solvent is standard dilution water.

[0095] Positive control: Swisse “Calcium&Vitamin D”, off-white tablets, batch number 33152A, Swisse, with the solvent being standard dilution water.

[0096] 1.2. Experimental animals

[0097] All zebrafish were raised in fish culture water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, with a conductivity of 450 - 550 μS / cm; pH of 6.5 - 8.5; hardness of 50 - 100 mg / L CaCO3). They were reproduced and provided by the fish breeding center of our company, with the experimental animal use license number: SYXK(Zhe)2022 - 0004. The feeding management meets the requirements of international AAALAC certification (certification number: 001458), and the IACUC ethics review number: IACUC - 2024 - 9219 - 01.

[0098] 1.3. Instruments, Consumables and Reagents

[0099] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); Electric focusing continuously variable fluorescence microscope (AZ100, Nikon, Japan); Precision electronic balance (CP214, OHAUS, USA); 6-well plate (Zhejiang Beilanbo Biotechnology Co., Ltd., China); Automatic sample rapid grinding instrument (JXFSTPRP-24L, Shanghai Jingxin Experimental Equipment Technology Department, China); Ultrasonic cleaner (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China).

[0100] Methyl cellulose (batch number B2006074, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Alizarin red (batch number C12071107, Shanghai Macklin Biochemical Co., Ltd., China); Anhydrous magnesium chloride (batch number E2322159, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Potassium hydroxide (batch number E2361048, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Glycerol (batch number H2220221, Shanghai Macklin Biochemical Co., Ltd., China); Anhydrous ethanol (batch number 20240426, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Hydrogen peroxide solution (batch number G2023089, Shanghai Aladdin Biochemical Technology Co., Ltd., China); 4% tissue cell fixative (batch number 240005013, Beijing Solarbio Science & Technology Co., Ltd., China); PBS phosphate buffer (dry powder) (batch number 18624437V, biosharp, China).

[0101] 2. Detection Methods

[0102] 2.1. Maximum Tested Concentration (MTC)

[0103] Randomly select wild-type AB strain zebrafish at 3 days post-fertilization (3 dpf) into beakers, with 30 zebrafish in each beaker. Administer the samples (concentrations are shown in Table 1) by water solution respectively, and the volume of each beaker is 20 mL. There are a total of 4 samples (2’-FL, 3’-SL, LNnT, and medium and long-chain fatty acid triglyceride (MLCT)), with 5 detection concentrations for each sample, and a normal control group is set simultaneously. Each group has 30 zebrafish. After treatment at 28 °C for 3 days, determine the MTC of the samples for normal zebrafish.

[0104] 2.2. Efficacy Evaluation of Single Sample in Promoting Bone Development

[0105] Randomly select 3dpf wild-type AB strain zebrafish into beakers, with 30 zebrafish treated in each beaker. Samples were dissolved in water and administered (concentrations are shown in Table 2). The positive control group was Swisse "Calcium & Vitamin D" at a concentration of 500 μg / mL. Meanwhile, a normal control group was set up, and the volume of each beaker was 20 mL. After treatment at 28 °C for 3 days, they were fixed and stained with alizarin red. Randomly select 10 zebrafish from each experimental group and place them under a fluorescence microscope for photographing. Use NIS-Elements D3.20 advanced image processing software to analyze and collect data, and analyze the fluorescence intensity of the zebrafish skull. Evaluate the efficacy of the sample in promoting bone 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.

[0106] 2.3. Evaluation of the Efficacy of the Composition in Promoting Bone Development

[0107] Randomly select 3dpf wild-type AB strain zebrafish into beakers, with 30 zebrafish treated in each beaker. Samples were dissolved in water and administered (concentrations are shown in Table 3), with a total of 25 groups. There were 21 ratios of the composition and 4 single samples. Meanwhile, the positive control group was Swisse "Calcium & Vitamin D" at a concentration of 500 μg / mL, and a normal control group was set up. The volume of each beaker was 20 mL. After treatment at 28 °C for 3 days, they were fixed and stained with alizarin red. Randomly select 10 zebrafish from each experimental group and place them under a fluorescence microscope for photographing. Use NIS-Elements D3.20 advanced image processing software to analyze and collect data, and analyze the fluorescence intensity of the zebrafish skull. Evaluate the efficacy of the sample in promoting bone 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.

[0108] 3. Detection Results

[0109] 3.1. MTC

[0110] Under the conditions of this experiment, the maximum test concentration (MTC) of 2FL and LNnT in promoting growth (bone) development was 2000 μg / mL; the MTC of MLCT in promoting growth (bone) development was 1500 μg / mL; the MTC of 3SL in promoting growth (bone) development was 1000 μg / mL, and the state of the zebrafish was not good. Therefore, the highest concentration for the subsequent experiment of 3SL was 500 μg / mL. See Table 1 for details.

[0111] Table 1. Results of the Experiment on Exploring the Concentration of the Sample in Promoting Growth (Bone) Development (n = 30)

[0112]

[0113] 3.2. Evaluation of the Efficacy of Single Samples in Promoting Bone Development

[0114] Under the conditions of this experiment, 2’-FL, LNnT, MLCT, and 3’-SL all have the efficacy of promoting bone development, specifically manifested as an increase in the fluorescence intensity of the skull. See Table 2 for details, Figure 1 and Figure 2 .

[0115] Table 2. Experimental Results of the Efficacy Evaluation of Single Samples in Promoting Bone Development (n = 10)

[0116]

[0117] Note: Compared with the normal control group, *p < 0.05, **p < 0.01, ***p < 0.001

[0118] Figure 1 Shows a typical image of the fluorescence intensity of the zebrafish skull after sample treatment. It can be seen from the figure that in the area marked by the dotted line, compared with the normal control group, the samples treated with 2’-FL, LNnT, MLCT, and 3’-SL all showed an increase in the fluorescence intensity of the skull, confirming the efficacy of promoting bone development.

[0119] Figure 2 Shows a bar chart of the fluorescence intensity of the zebrafish skull after sample treatment. This figure is based on the data in Table 2.

[0120] 3.3. Evaluation of the Efficacy of the Composition (MLCT + Human Milk Oligosaccharide) in Promoting Bone Development

[0121] The measurement results of the fluorescence intensity (bone density) of the zebrafish skull by the composition in this experiment are shown in Table 3 below.

[0122] Table 3. Fluorescence Intensity of the Skull Obtained by Compositions with Different Ratios

[0123]

[0124] Table 3 (continued). Fluorescence Intensity of the Skull Obtained by Compositions with Different Ratios

[0125]

[0126] Table 3 (continued). Fluorescence Intensity of the Skull Obtained by Compositions with Different Ratios

[0127]

[0128] Table 3 (continued). Fluorescence Intensity of the Skull Obtained by Compositions with Different Ratios

[0129]

[0130] Table 3 (continued). Fluorescence intensity of skulls obtained from compositions with different ratios

[0131]

[0132] 3.3.1. Analysis of experimental results

[0133] (1) MLCT + one type of human milk oligosaccharide

[0134] By analyzing the data in Table 3, the effects of MLCT and one type of human milk oligosaccharide on the fluorescence intensity (bone density) of zebrafish skulls are shown in Tables 4 - 6 below.

[0135] Table 4. Effects of MLCT + 2’-FL on the fluorescence intensity (bone density) of zebrafish skulls

[0136]

[0137] According to the results in Table 4, it can be seen that compared with MLCT alone (formulation 1), 2’-FL alone (formulation 2), and the combination of MLCT and 2’-FL with a ratio of 0.1:1 (formulation 14), the combinations of MLCT and 2’-FL with ratios in the range of 2:1 to 9:1 (formulations 20, 21, and 7) achieved a significant increase in the fluorescence intensity of the skull. This indicates that there is a synergistic effect in promoting bone development for the combination of MLCT and 2’-FL within this weight ratio range.

[0138] Table 5. Effects of MLCT + LNnT on the fluorescence intensity (bone density) of zebrafish skulls

[0139] Formulation No. Formulation 1 Formulation 16 Formulation 6 Formulation 23 Formulation 22 MLCT / Parts 1 0 2 5 9 2’-FL / Parts 0 0 0 0 0 LNnT / Parts 0 1 1 1 1 3’-SL / Parts 0 0 0 0 0 <![CDATA[Skull fluorescence intensity (pixels) (10 4 )]]> 121 93 144 132 146 <![CDATA[SE(10 4 )]]> 2 2 4 5 4

[0140] According to the results in Table 5, it can be seen that compared with MLCT alone (formulation 1) and LNnT alone (formulation 16), the combinations of MLCT and LNnT with ratios in the range of 2:1 to 9:1 (formulations 6, 23, and 22) achieved a significant increase in the fluorescence intensity of the skull. This indicates that there is a synergistic effect in promoting bone development for the combination of MLCT and LNnT within this weight ratio range.

[0141] Table 6. Effects of MLCT + 3’-SL on the fluorescence intensity (bone density) of zebrafish skulls

[0142] Formulation No. Formulation 1 Formulation 17 Formulation 25 Formulation 5 Formulation 24 MLCT / Parts 1 0 2 5 9 2’-FL / Parts 0 0 0 0 0 LNnT / Parts 0 0 0 0 0 3’-SL / Parts 0 1 1 1 1 <![CDATA[Skull fluorescence intensity (pixels) (10 4 )]]> 121 97 135 135 132 <![CDATA[SE(10 4 )]]> 2 4 3 4 4

[0143] According to the results in Table 6, it can be seen that compared with the individual MLCT (formulation 1) and individual 3'-SL (formulation 17), the combinations of MLCT and 3'-SL with a ratio of MLCT to 3'-SL in the range of 2:1 to 9:1 (formulations 25, 5, and 24) achieved a significant increase in the fluorescence intensity of the skull. This indicates that there is a synergistic effect in promoting bone development for the combinations of MLCT and 3'-SL within this weight ratio range.

[0144] (2) MLCT + two types of human milk oligosaccharides

[0145] By analyzing the data in Table 3, the effects of MLCT and two types of human milk oligosaccharides on the fluorescence intensity (bone density) of zebrafish skulls are shown in the following Tables 7 - 9.

[0146] Table 7. Effects of MLCT + (2'-FL + LNnT) on the fluorescence intensity (bone density) of zebrafish skulls

[0147] Formulation No. Formulation 1 Formulation 2 Formulation 16 Formulation 3 Formulation 8 MLCT / Parts 1 0 0 0 5 2’-FL / Parts 0 1 0 6 / 7 6 / 7 LNnT / Parts 0 0 1 1 / 7 1 / 7 3’-SL / Parts 0 0 0 0 0 <![CDATA[Skull fluorescence intensity (pixels) (10 4 )]]> 121 120 93 122 142 <![CDATA[SE(10 4 )]]> 2 2 2 3 5

[0148] According to the results in Table 7, it can be seen that compared with formulation 1 using individual MLCT, formulation 2 using individual 2'-FL, formulation 16 using individual LNnT, and formulation 3 using individual (2'-FL + LNnT), formulation 8 with the combination of MLCT and (2'-FL + LNnT) achieved a significant increase in the fluorescence intensity of the skull. This indicates that there is a synergistic effect in promoting bone development for the combination of MLCT and (2'-FL + LNnT) in formulation 8.

[0149] Table 8. Effects of MLCT + (LNnT + 3'-SL) on the fluorescence intensity (bone density) of zebrafish skulls

[0150] Formulation No. Formulation 1 Formulation 16 Formulation 17 Formulation 9 MLCT / Parts 1 0 0 6 2’-FL / Parts 0 0 0 0 LNnT / Parts 0 1 0 2 / 3 3’-SL / Parts 0 0 1 1 / 3 <![CDATA[Skull fluorescence intensity (pixels) (10 4 )]]> 121 93 97 149 <![CDATA[SE(10 4 )]]> 2 2 4 7

[0151] According to the results in Table 8, it can be seen that compared with formulation 1 using individual MLCT, formulation 16 using individual LNnT, and formulation 17 using individual 3'-SL, formulation 9 with the combination of MLCT and (LNnT + 3'-SL) achieved a significant increase in the fluorescence intensity of the skull. This indicates that there is a synergistic effect in promoting bone development for the combination of MLCT and (LNnT + 3'-SL) in formulation 9.

[0152] Table 9. Effects of MLCT + (2'-FL + 3'-SL) on the fluorescence intensity (bone density) of zebrafish skulls

[0153] Formulation No. Formulation 1 Formulation 2 Formulation 17 Formulation 10 MLCT / Parts 1 0 0 7 2’-FL / Parts 0 1 0 10 / 11 LNnT / Parts 0 0 0 0 3’-SL / Parts 0 0 1 1 / 11 <![CDATA[Skull fluorescence intensity (pixels) (10 4 )]]> 121 120 97 133 <![CDATA[SE(10 4 )]]> 2 2 4 5

[0154] According to the results in Table 9, it can be seen that compared with Formula 1 using only MLCT, Formula 2 using only 2’-FL, and Formula 17 using only 3’-SL, Formula 10 using MLCT in combination with (2’-FL + 3’-SL) achieved a significant increase in the fluorescence intensity of the skull. This indicates that there is a synergistic effect in the combination of MLCT and (2’-FL + 3’-SL) in Formula 10 in promoting bone development.

[0155] (3) MLCT + Three Types of Human Milk Oligosaccharides

[0156] By analyzing the data in Table 3, the effects of MLCT and three types of human milk oligosaccharides on the fluorescence intensity (bone density) of zebrafish skulls are shown in Table 10 below.

[0157] Table 10. Effects of MLCT + (2’-FL + LNnT + 3’-SL) on the Fluorescence Intensity (Bone Density) of Zebrafish Skulls

[0158]

[0159] According to the results in Table 10, it can be seen that compared with Formula 1 using only MLCT, Formula 2 using only 2’-FL, Formula 16 using only LNnT, Formula 17 using only 3’-SL, and Formulas 13 and 15 with a ratio of MLCT to (2’-FL + LNnT + 3’-SL) of 10:1 and 20:1, Formulas 11 and 12 with a ratio of MLCT to (2’-FL + LNnT + 3’-SL) of 1:1 and 3:1 achieved a significant increase in the fluorescence intensity of the skull, indicating that there is a synergistic effect in the combination of MLCT and (2’-FL + LNnT + 3’-SL) in Formulas 11 - 12 in promoting bone development.

[0160] The above description is only an exemplary embodiment of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the inventive concept of the present invention, improvements can still be made to the present invention, but these all fall within the protection scope of the present invention.

Claims

1. A nutritional composition, characterized in that It includes: Medium and long chain fatty acid triglycerides; and Human milk oligosaccharides, The weight ratio of the medium-chain fatty acid triglyceride to the human milk oligosaccharide is 1:1 to 9:

1. Wherein the human milk oligosaccharide is selected from one of the following: 2'-fucosyllactose; 3'-sialyl lactose; lactose-N-neotetraose; 2'-fucosyllactose and 3'-sialyl lactose; 2'-fucosyllactose and lactose-N-neotetraose; 3'-sialyl lactose and lactose-N-neotetraose; 2'-fucosyllactose and 3'-sialyl lactose and lactose-N-neotetraose.

2. The nutritional composition according to claim 1, characterized in that The weight ratio of the medium-chain fatty acid triglycerides to the human milk oligosaccharides is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1 or 9:

1.

3. The nutritional composition according to claim 1, characterized in that The mass ratio of the medium-chain fatty acid triglycerides to the human milk oligosaccharides is 2:1 to 9:

1.

4. The nutritional composition according to claim 1, characterized in that The human milk oligosaccharide is selected from one of the following: 2'-fucosyllactose; lacto-N-neotetraose; 2'-fucosyllactose and lacto-N-neotetraose.

5. The nutritional composition according to claim 1, characterized in that The human milk oligosaccharide is a combination of 2'-fucosyllactose, 3'-sialyl lactose and lactose-N-neotetraose, and the weight ratio of the medium-chain fatty acid triglyceride to the human milk oligosaccharide is 3:1 to 9:

1.

6. The nutritional composition according to claim 1, characterized in that The human milk oligosaccharide is any two selected from 2'-fucosyllactose, 3'-sialyl lactose and lactose-N-neotetraose, and the weight ratio of the medium-chain fatty acid triglyceride to the human milk oligosaccharide is 5:1 to 7:

1.

7. The nutritional composition according to claim 1, characterized in that The human milk oligosaccharide is 2'-fucosyllactose, and the weight ratio of the medium- and long-chain fatty acid triglycerides to the human milk oligosaccharide is 5:1 to 9:

1.

8. Food, characterized in that It comprises the nutritional composition according to any one of claims 1-7.

9. The food according to claim 8, characterized in that The food is infant food.

10. The food according to claim 8, characterized in that The food is selected from dairy products, candies, beverages, bread and biscuits.

11. The food according to claim 8, characterized in that The food is milk powder or fermented food.

12. The food according to claim 11, characterized in that The milk powder is infant formula milk powder.

13. The food according to any one of claims 8 to 12, characterized in that The content of the medium-chain fatty acid triglycerides is 0.1% to 30%, and the content of the human milk oligosaccharides is 0.01% to 6%, based on the weight of the food.

14. The food according to any one of claims 8 to 12, further comprising one or more selected from the group consisting of raw cow's milk, desalted whey powder, concentrated whey protein, lactose, edible vegetable blending oil, oligofructose, oligogalactose, nucleotides, choline, vitamins, minerals, DHA and taurine.

15. A medicament comprising the nutritional composition according to any one of claims 1 to 7.

16. Use of the nutritional composition according to any one of claims 1 to 7 or the food according to any one of claims 8 to 14 for the non-therapeutic purpose of promoting bone development.

17. The use according to claim 16, characterized in that The promoting bone development includes improving bone density.

18. The use according to any one of claims 16-17, characterized in that The promoting bone development includes promoting bone development in infants and / or children.

19. Use of the nutritional composition according to any one of claims 1 to 7 in the preparation of a medicine for promoting bone development.

20. The use according to claim 19, characterized in that The promoting bone development includes improving bone density.

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