Nutritional composition, food and pharmaceutical product comprising

Abstract

The present invention relates to nutritional compositions, food and pharmaceutical products comprising them, and uses thereof. The nutritional composition comprises medium-chain and long-chain fatty acid triglyceride and breast milk oligosaccharide, and the weight ratio of the medium-chain and long-chain fatty acid triglyceride to the breast milk oligosaccharide is 1: 1 to 10: 1. The nutritional composition disclosed by the invention can play a role in synergistically improving diarrhea.

Description

Technical Field

[0001] The present invention relates to the field of food, in particular to a nutritional composition, food and medicine comprising the nutritional composition, and uses thereof. Background Art

[0002] Diarrhea is a common digestive symptom and one of the main causes of malnutrition in children, especially in low-income countries or regions. Studies have shown that children under the age of 3 experience diarrhea three times a year on average, and each episode of diarrhea deprives children of the nutrients they need for growth. Although diarrhea may seem ordinary, its potential harm is widespread and serious.

[0003] There are many causes of diarrhea, including infectious factors and non-infectious factors. Among them, infectious factors include viral infections, such as those caused by viruses (such as rotavirus), bacteria (such as E. coli), parasites or fungi. These pathogens can directly damage the intestinal mucosa or secrete toxins to interfere with normal intestinal function; bacterial infections, such as salmonella, E. coli, etc., cause food poisoning through ingestion of contaminated food or water, leading to diarrhea; parasitic infections, such as cryptosporidiosis, amoebic dysentery, etc., are usually transmitted through drinking or using contaminated water. Non-infectious factors mainly include lactose intolerance, that is, due to the lack of sufficient lactase in the body, the lactose in dairy products cannot be effectively broken down, resulting in the fermentation of undigested lactose in the intestines, producing gas and acid, irritating the intestines, and causing diarrhea; drug side effects, such as some drugs may cause diarrhea; functional gastrointestinal diseases, such as irritable bowel syndrome, inflammatory bowel disease (such as Crohn's disease or ulcerative colitis), etc.

[0004] Lactose intolerance is an important non-infectious factor causing diarrhea. Lactose is a disaccharide whose molecules are composed of glucose and galactose. Lactose cannot be directly absorbed in the human body and needs to be broken down by lactase before it can be absorbed. However, some people lack lactase and are insufficient. After ingesting lactose, lactose cannot be digested and absorbed. The resulting reaction is called lactose intolerance, which is the condition of lactose intolerance. The main manifestations are abdominal distension, abdominal pain, diarrhea and other symptoms after eating milk and dairy products. The cause is related to genetics and small intestinal mucosal lesions. It can occur in any population, but is more common in children.

[0005] Therefore, it is desirable to provide a method for improving diarrhea, especially improving diarrhea in infants and / or 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] The weight ratio of the medium-chain fatty acid triglycerides to the human milk oligosaccharides is 1:1 to 10:1.

[0011] In a second aspect, the present invention relates to a food product comprising the nutritional composition according to the first aspect.

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

[0013] In a fourth aspect, the present invention relates to use of the nutritional composition according to the first aspect or the food according to the second aspect for the non-therapeutic purpose of improving diarrhea.

[0014] In a fifth aspect, the present invention relates to use of the nutritional composition according to the first aspect in preparing a medicament for improving diarrhea.

[0015] The inventors unexpectedly discovered during their research that when medium- and long-chain fatty acid triglycerides and human milk oligosaccharides are used in combination at a certain weight ratio, the two can act synergistically and significantly improve diarrhea and lactose intolerance diarrhea. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the technical solution of the present invention, the following will briefly describe the drawings required for describing the embodiments. It should be understood that these drawings are only for the convenience of technicians to understand the present invention more easily, and are not intended to limit the scope of the present invention.

[0017] Figure 1 Representative graphs showing the sum of opacity of zebrafish intestinal contents after single sample treatments.

[0018] Figure 2 Bar graph showing the sum of opacity of zebrafish intestinal contents after single sample treatment. Compared with the normal control group: * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001. DETAILED DESCRIPTION

[0019] In order to make the invention purpose, technical solution and beneficial technical effect of the present application clearer, the present application will be described in detail below. It should be noted that the various aspects, features, implementation methods and advantages described in the present application may be compatible and / or may be combined together.

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

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

[0022] In the present application, the term "medium-chain fatty acid" refers to fatty acids with 6-12 carbon atoms in the carbon chain, such as caproic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, etc.; the term "long-chain fatty acid" refers to fatty acids with more than 14 carbon atoms, generally 14-30 carbon atoms, in the carbon chain, 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 fatty acid triglyceride" and "long-chain fatty acid triglyceride" refer to the esterification reaction products of "medium-chain fatty acid" and "long-chain fatty acid" with glycerol, respectively; the term "medium-long chain fatty acid triglyceride" refers to triglycerides containing both medium-chain fatty acid residues and long-chain fatty acid residues in the molecular structure.

[0023] In the present application, the term "human milk oligosaccharides", also known as "human milk oligosaccharides", is a type of complex oligosaccharides composed of monosaccharides and their derivatives, sialic acid and other structural units connected by glycosidic bonds.

[0024] In the present application, the term "non-therapeutic purpose" has the conventional meaning in the art, which includes, for example, nutritional and / or health purposes, which can be used, for example, for preparing food such as ordinary food, functional food, health food or health products, etc.

[0025] The present invention relates to a nutritional composition, a food and a medicine comprising the nutritional composition, and uses thereof.

[0026] The present invention will be described in detail below.

[0027] Nutritional composition

[0028] In a first aspect, the present invention relates to a nutritional composition comprising medium-chain fatty acid triglycerides and human milk oligosaccharides, wherein the weight ratio of the medium-chain fatty acid triglycerides to the human milk oligosaccharides is 1: 1 to 10: 1. As an example, the mass ratio of the medium-chain fatty acid triglycerides to the human milk oligosaccharides may be 1: 1, 2: 1, 3: 1, 4: 1, 5: 1, 6: 1, 7: 1, 8: 1, 9: 1, 10: 1, or a range defined by any two of them.

[0029] The inventors unexpectedly discovered in their research that medium-chain fatty acid triglycerides and human milk oligosaccharides (particularly 2'-fucosyllactose, 3'-sialyllactose and / or lactose-N-neotetraose) can synergistically improve diarrhea, particularly lactose intolerance diarrhea, when used in combination in a specific ratio.

[0030] The present invention has no special 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 a pure form or in an impure form rich in human milk oligosaccharides.

[0031] Human milk oligosaccharides (also known as human milk oligosaccharides or human milk oligosaccharides) are the third largest solid component in breast milk, second only to lactose and fat, and have important biological activity. According to the representative groups, human milk oligosaccharides can be divided into human milk oligosaccharides containing fucosyl groups (such as 2'-fucosyllactose), human milk oligosaccharides containing sialic acid groups (such as 3'-sialyllactose) and non-fucosyl and non-sialic acid human milk oligosaccharides (such as lactose-N-neotetraose).

[0032] In some embodiments, the human milk oligosaccharide is selected from one or more of the following: fucosyl-containing human milk oligosaccharides, sialic acid-containing human milk oligosaccharides, and non-fucosyl and non-sialic acid human milk oligosaccharides. For example, the human milk oligosaccharides may be: one or more fucosyl-containing human milk oligosaccharides; one or more sialic acid-containing human milk oligosaccharides; one or more non-fucosyl and non-sialic acid human milk oligosaccharides; one or more fucosyl-containing human milk oligosaccharides, and one or more sialic acid-containing human milk oligosaccharides; one or more fucosyl-containing human milk oligosaccharides, and one or more non-fucosyl and non-sialic acid human milk oligosaccharides; one or more sialic acid-containing human milk oligosaccharides, and one or more non-fucosyl and non-sialic acid human milk oligosaccharides; one or more fucosyl-containing human milk oligosaccharides, one or more sialic acid-containing human milk oligosaccharides, and one or more non-fucosyl and non-sialic acid human milk oligosaccharides.

[0033] In some embodiments, the fucosyl-containing human milk oligosaccharide is, for example, selected from one or more of the following: 2'-fucosyllactose (2'-FL), 3-fucosyllactose, lactose-N-fucopentaose I, lactose-N-difucohexaose I, lactose-N-difucohexaose II, and lactose-bifucosyltetrose.

[0034] In some embodiments, the sialic acid-containing human milk oligosaccharide is, for example, selected from one or more of the following: 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL).

[0035] In some embodiments, the non-fucosyl non-sialic acid human milk oligosaccharide is, for example, selected from one or more of the following: lactose-N-tetraose (LNT), lactose-N-neotetraose (LNnT), lactose-N-triose II and lactose-N-diose.

[0036] In some embodiments, the human milk oligosaccharide is, for example, selected from one of the following: 2'-fucosyllactose; 3'-sialyllactose; lactose-N-neotetraose; 2'-fucosyllactose and 3'-sialyllactose; 2'-fucosyllactose and lactose-N-neotetraose; 3'-sialyllactose and lactose-N-neotetraose; 2'-fucosyllactose and 3'-sialyllactose and lactose-N-neotetraose.

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

[0038] When 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 taking into account 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.

[0039] For example, when the human milk oligosaccharide includes a fucosyl-containing human milk oligosaccharide (e.g., 2'-FL) and a sialic acid-containing human milk oligosaccharide (e.g., 3'-SL), the mass ratio of the fucosyl-containing human milk oligosaccharide / the sialic acid-containing human milk oligosaccharide, for example, 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, 1 1:1, 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.

[0040] For example, when the human milk oligosaccharide includes fucosyl-containing human milk oligosaccharide (e.g., 2'-FL) and non-fucosyl non-sialic acid human milk oligosaccharide (e.g., LNnT), the mass ratio of fucosyl-containing human milk oligosaccharide / non-fucosyl non-sialic acid human milk oligosaccharide, for example, the mass ratio of 2'-FL / LNnT can be 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 14:1, 15:1, 16:1, 17 ... :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.

[0041] For example, when the human milk oligosaccharide includes sialic acid-containing human milk oligosaccharide (e.g., 3'-SL) and non-fucosyl non-sialic acid-containing human milk oligosaccharide (e.g., LNnT), the mass ratio of sialic acid-containing human milk oligosaccharide to non-fucosyl non-sialic acid-containing human milk oligosaccharide is, for example, 3'-SL The mass ratio of LNnT may 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 oligosaccharide includes fucosyl-containing human milk oligosaccharide (e.g., 2'-FL), sialic acid-containing human milk oligosaccharide (e.g., 3'-SL) and non-fucosyl non-sialic acid human milk oligosaccharide (e.g., LNnT), the mass ratio of fucosyl-containing human milk oligosaccharide / sialic acid-containing human milk oligosaccharide, e.g., 2'-FL / 3'-SL mass ratio, the mass ratio of fucosyl-containing human milk oligosaccharide / non-fucosyl non-sialic acid human milk oligosaccharide, e.g., 2'-FL / LNnT mass ratio, and the mass ratio of sialic acid-containing human milk oligosaccharide / non-fucosyl non-sialic acid human milk oligosaccharide, e.g., 3'-SL The mass ratio of LNnT 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. For example, when the human milk oligosaccharide is a combination of fucosyl-containing human milk oligosaccharides, sialic acid-containing human milk oligosaccharides and non-fucosyl non-sialic acid human milk oligosaccharides, based on 4 parts by weight of non-fucosyl non-sialic acid human milk oligosaccharides (e.g. LNnT), the amount of fucosyl-containing human milk oligosaccharides (e.g. 2'-FL) ranges from 1 to 20, preferably 8 to 28 parts by weight, and / or the amount of sialic acid-containing human milk oligosaccharides (e.g. 3'-SL) ranges from 1 to 8, preferably 1 to 4 parts by weight.

[0043] Medium-chain triglycerides (MLCT) are structural lipids that contain both medium-chain fatty acids and long-chain fatty acids in the triglyceride molecule. They have different physical and chemical properties, metabolic characteristics and nutritional value from long-chain triglycerides (LCT) or medium-chain triglycerides (MCT). Studies (see, for example, Yuan Tinglan, Medium-chain triglyceride composition and metabolic characteristics of breast milk fat [D], Jiangnan University, 2021) show that medium-chain triglycerides (MLCT) are not simply equivalent to a physical mixture of long-chain triglycerides (LCT) and medium-chain triglycerides (MCT) (of course, they are not equivalent to a simple mixture of long-chain fatty acids and medium-chain fatty acids), and the latter does not have the physical and chemical properties, metabolic characteristics and nutritional value of the former.

[0044] The present invention has no special requirements for the medium-chain triglycerides used, and the medium-chain triglycerides commonly used in the art can be used. The medium-chain triglycerides can be used in pure form or in an impure form rich in medium-chain triglycerides. In some embodiments, the medium-chain triglycerides of the present invention contain C6-C 12 (e.g. C6, C7, C8, C9, C 10 , C 11 , C 12 , or the range defined by any two thereof) 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 thereof) fatty acid residues, wherein:

[0045] The C6-C 12 The fatty acid residue is derived from one or more of the following: caproic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecanoic acid, dodecanoic acid, etc., preferably caproic acid, caprylic acid, capric acid and lauric acid; and / or

[0046] The C 14 -C 30 The fatty acid residue is derived from one or more of the following: myristic acid, palmitic acid, heptadecanoic acid, oleic acid, linoleic acid, stearic acid, nonadecanoic acid, eicosapentaenoic acid, heneicosanoic acid, docosatetraenoic acid, docosahexaenoic acid, tricosanoic acid, tetracosenoic acid, eicosapentaenoic acid, hexacoshexaenoic 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.

[0047] In some embodiments, the C6-C 12 The fatty acid residues on the C 14 -C 30 The weight ratio of fatty acid residues is 0.124 to 2.000. As an example, the C6-C 12The fatty acid residues on the C 14 -C 30 The weight ratio of the fatty acid residues may 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.

[0048] In some embodiments, the nutritional composition of the present invention may consist of medium- and long-chain fatty acid esters and human milk oligosaccharides.

[0049] food

[0050] In a second aspect, the present invention relates to a food product comprising the nutritional composition according to the first aspect.

[0051] In some embodiments, the food is an infant food such as an infant formula.

[0052] In some embodiments, based on the weight of the food, the content of medium and long-chain fatty acid triglycerides can be 0.1 to 30% by weight. As an example, based on the weight of the food, the content of medium and long-chain fatty acid 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.

[0053] In some embodiments, based on the weight of the food, the content of human milk oligosaccharides may be 0.01 to 20% by weight. As an example, based on the weight of the food, the content of human milk oligosaccharides may 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, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20% by weight, or within the range defined by any two of them.

[0054] In some embodiments, the food may be, for example, a functional food, a health food, or a health product.

[0055] In some embodiments, the food is a common food or a special food. As an example, the food can be selected from dairy products, candies, beverages, bread and biscuits, for example, the food can be selected from milk powder or fermented food.

[0056] In some embodiments, the milk powder is infant formula milk powder.

[0057] In addition, it is easy for those skilled in the art to understand that, in addition to the nutritional composition of the first aspect of the present invention, the food also includes one or more selected from the following: raw milk, desalted whey powder, concentrated whey protein, lactose, edible vegetable blending oil, oligofructose, oligogalactose, nucleotides, choline, vitamins, minerals, DHA and taurine.

[0058] For example, when the food is milk powder, in addition to the nutritional composition, the milk powder may also include proteins such as α-lactalbumin, milk fat globule membrane protein; carbohydrates such as lactose; lipids; minerals such as calcium, iron, phosphorus, etc.; vitamins; other additives such as whey powder, choline bitartrate, docosahexaenoic acid, arachidonic acid, walnut oil, etc.

[0059] The food can 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 described in detail here.

[0060] All the above descriptions about the nutritional composition of the first aspect of the present invention are applicable here and will not be repeated here.

[0061] drug

[0062] In a third aspect, the present invention relates to a medicament comprising the nutritional composition according to the first aspect.

[0063] The present invention has no special requirements for the dosage form of the drug, and the commonly used dosage forms of drugs in the art can be used. As an example, the drug of the present invention can be a solid preparation, such as powder, granules, tablets, capsules, drop pills, pills, etc.; a semi-solid preparation, such as an ointment, cream, eye ointment, gel, etc.; a liquid preparation, such as an oral liquid preparation, such as a syrup, mixture, suspension, emulsion, etc.

[0064] It is easy for those skilled in the art to understand that, depending on the dosage form and application scenario of the drug, the drug may also include various excipients permitted in pharmacology. In addition, the drug of the present invention may be prepared by methods commonly used in the art, which is well known to those skilled in the art and will not be described in detail here.

[0065] All the above descriptions about the nutritional composition of the first aspect of the present invention are applicable here and will not be repeated here.

[0066] use

[0067] In a fourth aspect, the present invention relates to 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.

[0068] The improving diarrhea may, for example, include improving lactose intolerance type diarrhea.

[0069] The improvement of diarrhea may, for example, include, but is not limited to, improvement of diarrhea in infants and / or children and / or adolescents.

[0070] The non-therapeutic purpose may include, for example, nutritional and / or health purposes, such as for preparing food such as ordinary food, functional food, health food or health care products.

[0071] In some embodiments, when the nutritional composition or food is used for oral administration by humans, the dosage is such that the dosage of the medium-chain fatty acid triglyceride is 0.1 to 30 g / day, preferably 2.5 to 30 g / day. As an example, the dosage of the medium-chain fatty acid triglyceride may 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.

[0072] In some embodiments, when the nutritional composition or food is used for oral administration to humans, the dosage is such that the dosage of the human milk oligosaccharide is 0.05 to 25 g / day, preferably 0.1 to 25 g / day. As an example, the dosage of the human milk oligosaccharide may 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, the dosage of the medium and long chain fatty acid triglycerides may be converted according to the ratio with human milk oligosaccharides.

[0073] In a fifth aspect, the present invention relates to use of the nutritional composition according to the first aspect in preparing a medicament for improving diarrhea.

[0074] All the above descriptions about the nutritional composition of the first aspect, the food of the second aspect and the medicine of the third aspect of the present invention are applicable here and will not be repeated here.

[0075] Example

[0076] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the 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.

[0077] Experimental Principle

[0078] In this experiment, a lactose-induced zebrafish lactose intolerance diarrhea model was used.

[0079] Compared with traditional animal experiments (piglets, mice and rats) and human experiments, zebrafish experimental schemes have many advantages in the evaluation of the efficacy of promoting growth (bone) development. The zebrafish experimental scheme has many advantages in the evaluation of the efficacy of improving diarrhea. Compared with traditional animal experiments (such as piglets, mice and rats) and human experiments, it is specifically reflected in the following aspects: (1) Fast and efficient: The zebrafish model has a shorter experimental cycle, is easy to operate, and the results are easy to observe and the cost is low. This makes zebrafish an efficient living animal model that can achieve early, rapid, and high-throughput screening of drugs that promote intestinal function activity. (2) High safety: The zebrafish model can effectively increase the speed of drug screening while ensuring the safety of the screened drugs. This is because the physiology, development, and metabolism of zebrafish are highly similar to those of mammals, and the homology of their genes with human genes is 87%, which can reliably simulate and predict human physiological and pathological processes. (3) High-throughput screening: The zebrafish model can achieve high-throughput screening, which is very beneficial for early testing and evaluation of candidate drugs during the development of new drugs. In addition, the application of the zebrafish model can also save a lot of experimental costs. (4) Similar intestinal composition: The zebrafish intestine is similar to that of humans, consisting of endothelial cells, connective tissue, circular muscles and external longitudinal muscles, which gives it a unique advantage in simulating human intestinal function. (5) Standardized procedures: The first complete zebrafish technical procedure for food health function testing in China has been published and has been recognized and supported by authoritative scientific research institutes and enterprises. These procedures provide scientific and rigorous technical guidance for zebrafish experiments, ensuring the accuracy and reproducibility of experimental results. In short, the zebrafish intestine is highly homologous to the mammalian intestine in development, organization and function. The gene expression and transcriptional regulation in intestinal epithelial cells are highly conserved with mammals, and the metabolic function is also conserved. The zebrafish digestive system basically functions 5 days after fertilization, making it an important model organism for studying the basic processes of intestinal inflammation and injury.

[0080] Lactose intolerance (LI) refers to abdominal and systemic symptoms such as abdominal pain, bloating and diarrhea that occur after lactose is ingested by people with lactose malabsorption. A small number of patients may experience headaches and nausea. Lactose is a type of short-chain fermentable carbohydrates that can cause symptoms through similar mechanisms. LI accounts for 69% of the total food intolerance population, ranking first, and 56.6% of them have multiple food intolerances at the same time, which seriously affects the quality of life of patients. Typical symptoms of lactose intolerance include abdominal pain, bloating, flatulence, diarrhea, and less nausea and vomiting. The mechanism and time correlation of systemic symptoms remain controversial. Abdominal pain and bloating are caused by the fermentation of unabsorbed lactose in the colon. Intestinal flora provides a salvage pathway for lactose digestion by breaking down lactose into short-chain fatty acids and gas. Lactose that is not absorbed in the ileum and colon leads to acidification of colonic contents and an increase in osmotic load. This will lead to increased secretion of electrolytes and fluids, as well as rapid transit time, leading to the appearance of loose stools and diarrhea. The zebrafish intestine is directly connected to the esophagus. It is a compartmentalized tubular structure divided into three regions: the foregut, midgut, and hindgut. The zebrafish foregut, also known as the foregut bulb, is larger than the midgut and hindgut, and can act as a storage device equivalent to the stomach. The zebrafish intestine is highly homologous to the mammalian intestine in development, organization, and function. Gene expression and transcriptional regulation in intestinal epithelial cells are highly conserved with mammals, and metabolic functions are also conserved. The zebrafish digestive system is basically functional 5 days after fertilization, making it an important model organism for studying the basic processes of intestinal inflammation and injury.

[0081] The sources and specifications of the raw materials used in the following examples are as follows:

[0082] Medium-chain fatty acid triglycerides: derived from medium-chain fatty acid edible oil (provided by Qingdao Haizhiyuan Life Science Technology Co., Ltd., batch number Y1501-23070101), purity 69.8%;

[0083] 2'-fucosyllactose: provided by DSM Vitamins Trading (Shanghai) Co. Ltd. SHANGHAI, purity 99.2%, batch number: 21107002;

[0084] 3'-Sialyl lactose: provided by DSM Vitamins Trading (Shanghai) Co. Ltd. SHANGHAI, purity 95.2%, batch number: DK21294101;

[0085] Lactose-N-neotetraose: provided by DSM Vitamins Trading (Shanghai) Co. Ltd. DTL DutyPaid. Shanghai, purity 100.5%, batch number: 20454001;

[0086] Wild-type AB strain zebrafish: bred and provided by the fish breeding center of Hangzhou Huante Biotechnology Co., Ltd., with the experimental animal use license number: SYXK (Zhejiang) 2022-0004.

[0087] Positive control drug

[0088] Galacto-oligosaccharides.

[0089] In the following examples, AB strain zebrafish were used for animal experiments.

[0090] 1. Test materials

[0091] 1.1. Sample preparation information

[0092] For MLCT, 2'-FL, LNnT and 3'-SL, the solvent was standard dilution water.

[0093] Positive control: galacto-oligosaccharide, white powder, batch number C15916553, Shanghai MacLean Biochemical Technology Co., Ltd., solvent is standard dilution water.

[0094] 1.2. Experimental animals

[0095] Zebrafish were raised in fish farming water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, conductivity was 450-550 μS / cm; pH was 6.5-8.5; hardness was 50-100 mg / L CaCO3), bred and provided by our company's fish farming center, with the experimental animal use license number: SYXK (Zhejiang) 2022-0004, and the breeding management complies with the requirements of the international AAALAC certification (certification number: 001458), and the IACUC ethics review number: IACUC-2024-10070-01.

[0096] 1.3. Instruments, consumables and reagents

[0097] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); precision electronic balance (CP214, OHAUS, USA); 6-well plate (Zhejiang Bellambo Biotechnology Co., Ltd., China); ultrasonic cleaning machine (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China).

[0098] Lactose, anhydrous (Batch No. D2328302, Shanghai Aladdin Biochemical Technology Co., Ltd., China); egg yolk powder (Batch No. 20230203, Zhejiang Aige Biotechnology Co., Ltd., China).

[0099] 2. Detection Methods

[0100] 2.1. Determination of maximum detectable concentration (MTC)

[0101] Wild-type AB zebrafish at 4 days post-fertilization (dpf) were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). The samples were given in water (concentrations shown in Table 1), and a normal control group and a model control group were set up at the same time, with a volume of 3 mL per well. After 1 day of treatment at 28°C, the samples were washed away, and except for the normal control group, the other concentration groups were fed with lactose and egg yolk powder in water to establish a lactose intolerance model. After 4 hours of treatment at 28°C, the lactose and egg yolk powder were washed away, and the samples were left to stand for 2 hours to measure the MTC of the model zebrafish.

[0102] 2.2. Evaluation of single sample efficacy in improving diarrhea

[0103] 4 dpf wild-type AB strain zebrafish were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). The samples were given water-soluble (concentrations are shown in Table 2), and the positive control galacto-oligosaccharide concentration was 2000 μg / mL. At the same time, a normal control group and a model control group were set up, and the volume of each well was 3 mL. After 1 day of treatment at 28℃, the samples were washed away. Except for the normal control group, the zebrafish in the other concentration groups were fed with lactose and egg yolk powder in water to establish a lactose intolerance model. After 4 hours of treatment at 28℃, the lactose and egg yolk powder were washed away and left to stand for 2 hours. Ten zebrafish were randomly selected from each experimental group and photographed under a dissecting microscope. The NIS-ElementsD3.20 advanced image processing software was used to analyze and collect data. The sum of the opacity of the zebrafish intestinal contents was analyzed, and the statistical analysis results of this indicator were used to evaluate the efficacy of the samples in improving lactose intolerance (diarrhea). The statistical analysis results are expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0104] 2.3. Evaluation of the efficacy of the composition in improving diarrhea

[0105] 4 dpf wild-type AB strain zebrafish were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). The samples were given water-soluble (concentrations are shown in Table 3), the positive control galacto-oligosaccharide concentration was 2000 μg / mL, and the normal control group and model control group were set up at the same time, with a volume of 3 mL per well. After 1 day of treatment at 28℃, the samples were washed away. Except for the normal control group, the zebrafish in the other concentration groups were fed with lactose and egg yolk powder in water-soluble to establish a lactose intolerance model. After 4 hours of treatment at 28℃, the lactose and egg yolk powder were washed away and left to stand for 2 hours. Ten zebrafish were randomly selected from each experimental group and photographed under a dissecting microscope. The NIS-ElementsD3.20 advanced image processing software was used to analyze and collect data. The sum of the opacity of the zebrafish intestinal contents was analyzed, and the statistical analysis results of this indicator were used to evaluate the efficacy of the samples in improving lactose intolerance (diarrhea). The statistical analysis results are expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0106] 3. Test results

[0107] 3.1. MTC

[0108] Under the experimental conditions, the MTCs of MLCT, 2'-FL, LNnT and 3'-SL in improving diarrhea were 2000 μg / mL, 2000 μg / mL, 2000 μg / mL and 1000 μg / mL, respectively. See Table 1 for details.

[0109] Table 1. Maximum detection concentration (MTC) experimental results of samples (n = 30)

[0110]

[0111] From the results in Table 1, it can be seen that under the experimental conditions, when the maximum detection concentration (MTC) of MLCT, 2'-FL, and LNnT was 2000 μg / mL, the state of zebrafish was similar to that of the model control group; when the treatment concentration of 3'-SL was 1500 μg / mL, the state was worse than that of the model control group, indicating that this concentration was intolerant, and at 1000 μg / mL, the state of zebrafish was similar to that of the model control group. Therefore, the concentration of 3'-SL used in subsequent experiments did not exceed 1000 μg / mL.

[0112] 3.2. Efficacy of single sample in improving diarrhea

[0113] Under the experimental conditions, MLCT, 2'-FL, LNnT and 3'-SL all have the effect of improving diarrhea, specifically improving diarrhea caused by lactose intolerance. See Table 2 for details. Figure 1 and Figure 2 .

[0114] Table 2. Results of single sample diarrhea improvement efficacy test (n = 10)

[0115]

[0116] Compared with the model control group, **p < 0.01, ***p < 0.001

[0117] Figure 1 A typical graph of the sum of intestinal content opacity of zebrafish after single sample treatment is shown. As can be seen from the figure, in the area marked with dotted lines, compared with the normal control group, the samples treated with 2'-FL, LNnT, MLCT and 3'-SL all showed a significant increase in the sum of intestinal content opacity, confirming the effect of improving diarrhea.

[0118] Figure 2 A bar graph showing the sum of the opacity of zebrafish intestinal contents after single sample treatments is shown. This graph is based on the data in Table 2.

[0119] 3.3. Composition for improving diarrhea

[0120] The results of the measurement of the composition on the sum of the opacity of zebrafish intestinal contents are shown in Table 3 below.

[0121] Table 3. The sum of intestinal content opacity obtained by the compositions with different ratios

[0122] Note: The amount of each formula in the table is expressed as follows: MLCT+2'-FL+LNnT+3'-SL (μg / mL) = total amount, the same below.

[0123] Table 3 (continued). The sum of intestinal content opacity obtained by the compositions with different ratios

[0124] Table 3 (continued). The sum of intestinal content opacity obtained by the compositions with different ratios

[0125] Table 3 (continued). The sum of intestinal content opacity obtained by the compositions with different ratios

[0126] 3.3.1. Experimental results analysis

[0127] To facilitate the analysis and explanation of the results, the results in Table 3 are further split and combined into Tables 4 to 11.

[0128] (1) MLCT + a type of human milk oligosaccharide

[0129] The effects of MLCT and one type of human milk oligosaccharide on the sum of opacity (pixels) of zebrafish intestinal contents are summarized in the following Tables 4-7.

[0130] Table 4. Effects of MLCT + 2'-FL on the sum of opacity of zebrafish intestinal contents

[0131]

[0132] According to the results in Table 4, it can be seen that the combination of MLCT and 2'-FL (Formulation 18, Formulation 19 and Formulation 7) with a ratio of MLCT to 2'-FL of 1:1 to 9:1 achieved a significant improvement in the sum of intestinal content opacity compared to MLCT alone (Formulation 1) and 2'-FL alone (Formulation 2), which indicates that the above combination of MLCT and 2'-FL has a synergistic effect in improving diarrhea.

[0133] Table 5. Effects of MLCT + LNnT on the sum of opacity of zebrafish intestinal contents (total concentration 400 μg / mL)

[0134]

[0135] According to the results in Table 5, it can be seen that compared with MLCT alone (Formula 1), LNnT alone (Formula 16) and a combination of MLCT and LNnT with a ratio of 0.1:1 (Formula 14), the combination of MLCT and LNnT with a ratio of MLCT to LNnT of 1:1 to 9:1 (Formula 6, Formula 21 and Formula 20) achieved a significant improvement in the sum of intestinal content opacity, indicating that the combination of MLCT and LNnT in Formula 6, Formula 21 and Formula 20 has a synergistic effect in improving diarrhea.

[0136] Table 6. Effects of MLCT + LNnT on the sum of opacity of zebrafish intestinal contents (total concentration 1000 μg / mL)

[0137]

[0138] According to the results in Table 6, it can be seen that the combination of MLCT and LNnT (Formulation 24, Formulation 26 and Formulation 27) with a ratio of MLCT to LNnT ranging from 1:1 to 9:1 achieved a significant improvement in the sum of intestinal content opacity compared to MLCT alone (Formulation 28) and LNnT alone (Formulation 25), indicating that the combination of MLCT and LNnT has a synergistic effect in improving diarrhea.

[0139] Therefore, it can be seen from Tables 5-6 that at different dosages, the combination of MLCT and LNnT has a synergistic effect in improving diarrhea.

[0140] Table 7. Effects of MLCT + 3'-SL on the sum of opacity of zebrafish intestinal contents

[0141]

[0142] According to the results in Table 7, it can be seen that the combination of MLCT and 3'-SL (Formula 23, Formulation 5 and Formulation 22) with a ratio of MLCT to 3'-SL of 1:1 to 9:1 achieved a significant improvement in the sum of intestinal content opacity compared to MLCT alone (Formulation 1) and 3'-SL alone (Formulation 17), indicating that the combination of MLCT and 3'-SL has a synergistic effect in improving diarrhea.

[0143] (2) MLCT + two types of human milk oligosaccharides

[0144] The effects of MLCT and two types of human milk oligosaccharides on the sum of opacity (pixels) of zebrafish intestinal contents are summarized in the following Tables 8-10.

[0145] Table 8. Effects of MLCT + (2'-FL+LNnT) on the sum of opacity of zebrafish intestinal contents

[0146]

[0147] According to the results in Table 8, it can be seen that the combination of MLCT and (2'-FL+LNnT) (Formula 8) with a ratio of MLCT to (2'-FL+LNnT) of 5:1 achieved a significant improvement in the sum of intestinal content opacity compared to MLCT alone (Formula 1), 2'-FL alone (Formula 2), LNnT alone (Formula 16), and (2'-FL+LNnT) alone (Formula 3). This indicates that the above combination of MLCT and (2'-FL+LNnT) has a synergistic effect in improving diarrhea.

[0148] Table 9. Effects of MLCT + (LNnT+3'-SL) on the sum of opacity of zebrafish intestinal contents

[0149]

[0150] According to the results of Table 9, it can be seen that the combination of MLCT and (LNnT+3'-SL) (Formula 9) with a ratio of MLCT to (LNnT+3'-SL) of 6:1 achieved a significant improvement in the sum of intestinal content opacity compared to MLCT alone (Formula 1), LNnT alone (Formula 16), and 3'-SL alone (Formula 17). This indicates that the above combination of MLCT and (LNnT+3'-SL) has a synergistic effect in improving diarrhea.

[0151] Table 10. Effects of MLCT + (2'-FL+3'-SL) on the total opacity of zebrafish intestinal contents

[0152]

[0153] According to the results in Table 10, it can be seen that the combination of MLCT and (2'-FL+3'-SL) (Formula 10) with a ratio of MLCT to (2'-FL+3'-SL) of 7:1 achieved a significant improvement in the sum of intestinal content opacity compared to MLCT alone (Formula 1), 2'-FL alone (Formula 2), and 3'-SL alone (Formula 17). This indicates that the above combination of MLCT and (2'-FL+3'-SL) has a synergistic effect in improving diarrhea.

[0154] (3) MLCT + three types of human milk oligosaccharides

[0155] The effects of MLCT and three types of human milk oligosaccharides on the sum of opacity (pixels) of zebrafish intestinal contents are summarized in Table 11 below.

[0156] Table 11. Effects of MLCT + (2'-FL+LNnT+3'-SL) on the total opacity of zebrafish intestinal contents

[0157]

[0158] According to the results of Table 11, it can be seen that the combination of MLCT and (2'-FL+LNnT+3'-SL) (Formula 11 and Formula 12) with a ratio of MLCT to (2'-FL+LNnT+3'-SL) of 1:1 to 10:1 achieved a significant improvement in the sum of intestinal content opacity compared to MLCT alone (Formula 1), 2'-FL alone (Formula 2), LNnT alone (Formula 16), and 3'-SL alone (Formula 17). This indicates that the above combination of MLCT and (2'-FL+LNnT+3'-SL) has a synergistic effect in improving diarrhea.

[0159] The above description is only an exemplary embodiment of the present invention. It should be noted that, for those skilled in the art, without departing from the creative concept of the present invention, the present invention can also be improved, but these all belong to 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 triglycerides to the human milk oligosaccharides is 1:1 to 10:

1.

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, 9:1 or 10:

1.

3. The nutritional composition according to claim 1, characterized in that The human milk oligosaccharide is selected from one or more of the following: human milk oligosaccharides containing fucosyl, human milk oligosaccharides containing sialic acid, and human milk oligosaccharides containing neither fucosyl nor sialic acid.

4. The nutritional composition according to claim 3, characterized in that The fucosyl-containing human milk oligosaccharide is selected from one or more of the following: 2'-fucosyllactose, 3-fucosyllactose, lactose-N-fucopentaose I, lactose-N-difucohexaose I, lactose-N-difucohexaose II, and lactose-bifucosyltetrose.

5. The nutritional composition according to claim 3, characterized in that The sialic acid-containing human milk oligosaccharide is selected from one or more of the following: 3'-sialyllactose and 6'-sialyllactose.

6. The nutritional composition according to claim 3, characterized in that The non-fucosyl and non-sialic acid human milk oligosaccharides are selected from one or more of the following: lactose-N-tetraose, lactose-N-neotetraose, lactose-N-triose II and lactose-N-disaccharide.

7. The nutritional composition according to claim 1, characterized in that The human milk oligosaccharide is selected from 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.

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

9. The nutritional composition according to any one of claims 1 to 8, 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- and long-chain fatty acid triglycerides to the human milk oligosaccharide is 1:1 to 3:

1.

10. Food comprising the nutritional composition according to any one of claims 1 to 9.

11. The food according to claim 10, characterized in that The food is infant food.

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

13. The food according to claim 10, characterized in that The food is milk powder or fermented food.

14. The food according to claim 13, characterized in that The milk powder is infant formula milk powder.

15. The food according to any one of claims 10 to 14, 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 20%, based on the total dry weight of the food.

16. The food according to any one of claims 10 to 14, 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.

17. A medicament comprising the nutritional composition according to any one of claims 1 to 9.

18. Use of the nutritional composition according to any one of claims 1 to 9 or the food according to any one of claims 10 to 16 for the non-therapeutic purpose of improving diarrhea.

19. The use according to claim 18, characterized in that The diarrhea includes lactose intolerance type diarrhea.

20. The use according to any one of claims 18-19, characterized in that The improving diarrhea includes improving diarrhea in infants and / or children.

21. Use of the nutritional composition according to any one of claims 1 to 9 in the preparation of a medicament for improving diarrhea.

Images