A nutritional composition for improving gut function

Abstract

The application discloses a kind of nutrition composition for improving intestinal function.The application provides a kind of nutrition composition for improving intestinal function, it includes iso-C15:0 and anteiso-C15:0;The content of the iso-C15:0 is compared with the content of the anteiso-C15:0 in mass ratio is 1:(0.1-10);Wherein, the nutrition composition further includes 3'-SL, the content of the iso-C15:0 and the anteiso-C15:0 is compared with the content of the 3'-SL in mass ratio is 1:(0.01-20).The nutrition composition provided by the application includes branched chain fatty acid iC15:0 and aC15:0, has the effect of improving intestinal function, and produces synergistic effect, specifically has the efficacy of slowing down the degree of intestinal damage, balancing intestinal inflammation, strengthening intestinal barrier function.

Description

Technical Field

[0001] This invention belongs to the field of nutrient research, and specifically relates to a nutritional composition for improving intestinal function. Background Technology

[0002] Intestinal inflammation is a group of digestive tract diseases or non-pathological states characterized by intestinal mucosal immune imbalance, barrier damage, microecological dysbiosis, and chronic low-grade inflammation. It mainly includes inflammatory bowel diseases (ulcerative colitis, Crohn's disease), as well as infectious enteritis, nonspecific chronic enteritis, and irritable bowel syndrome with inflammation. With changes in dietary structure, increased life stress, and environmental factors, the incidence of intestinal inflammation is rising year by year, exhibiting characteristics such as prolonged course, recurrent attacks, high risk of complications, and serious impact on quality of life, making it a significant global public health issue.

[0003] The development and progression of intestinal inflammation are closely related to multiple mechanisms: disruption of the intestinal mucosal barrier integrity leads to increased intestinal permeability; endotoxins and antigen translocation trigger excessive activation of innate and adaptive immunity; pro-inflammatory factors are released in large quantities, forming an inflammatory cascade; simultaneously, intestinal flora imbalance occurs, with decreased abundance of beneficial bacteria, excessive proliferation of pathogenic and opportunistic pathogens, and insufficient protective metabolites such as short-chain fatty acids, further exacerbating inflammation and mucosal damage. Both clinical and basic research have confirmed that nutritional intervention can not only improve patients' nutritional status but also achieve safe, gentle, and sustained improvement of intestinal inflammation through multi-target and multi-pathway synergistic effects, including regulating intestinal flora, repairing the mucosal barrier, modulating inflammatory pathways, and clearing oxidative stress damage. Therefore, developing safe, effective, highly compliant, and long-term usable nutritional compositions has become an important direction for the management and adjunctive intervention of intestinal inflammation.

[0004] Existing technologies employ various methods to intervene in intestinal inflammation, but they suffer from the following drawbacks: 1. Significant side effects from drug treatments and insufficient safety for long-term use: Traditional treatments primarily utilize aminosalicylic acids, glucocorticoids, immunosuppressants, and biological agents. While these can quickly control acute inflammation, long-term use can easily lead to adverse reactions such as gastrointestinal discomfort, liver and kidney damage, bone marrow suppression, increased risk of infection, osteoporosis, and abnormal blood sugar and lipids. Biological agents are expensive and suffer from problems such as primary non-response, secondary ineffectiveness, and antibody production, making them unsuitable as long-term maintenance and daily care methods. 2. The effects of common probiotics vary from person to person, with different individuals experiencing different symptoms. Furthermore, the activity and stability of probiotics significantly reduce their effectiveness after passing through the gastrointestinal tract. Common prebiotics can cause excessive gas production in the human body, clinically manifesting as bloating. 3. Poor stability and compliance, making long-term use difficult: Active ingredients such as probiotics are easily deactivated by temperature, humidity, and pH, resulting in a short shelf life. Some products have poor taste and are inconvenient to take, leading to low long-term patient compliance and failing to meet the clinical need for continuous intervention in chronic inflammation.

[0005] Branched-chain fatty acids are saturated fatty acids with one or more methyl branches on their carbon chains. The main configuration is a single methyl branch at the end of the carbon chain. When the methyl branch is located on the second carbon atom at the alkyl end, it is called iso-BCFA; when it is located on the third carbon atom at the alkyl end, it is called anteiso-BCFA.

[0006] Previous studies and patents have confirmed the efficacy of branched-chain fatty acids (BCFAs) in inhibiting intestinal inflammation. For example, reference 1 (CN118416112A) discloses a composition containing branched-chain fatty acids such as 14-methylpentadecanoic acid, 12-methyltridecanoic acid, and 13-methyltetradecanoic acid. This composition can inhibit ROS generation in a LPS-induced calf small intestinal epithelial cell inflammation model; and it can regulate the levels of inflammatory factors in the intestines of DSS-induced colitis mice and improve the gastrointestinal health of mice. Reference 2 (WO2023031226A1) discloses the addition of branched-chain fatty acids to a CaCo2 cell culture system and the gavage administration of branched-chain fatty acids to DSS-induced colitis mice. The results show that BCFAs can induce high SUMOylation of intestinal proteins in CaCo2 cells, thereby reducing the expression of pro-inflammatory cytokines. In addition, BCFAs also inhibit DSS-induced intestinal inflammation, reduce the activity of intestinal cell deaminases, and promote intestinal epithelial integrity. Similarly, reference 3 (CN115300530A) also discloses that branched-chain fatty acids have the function of inhibiting intestinal inflammation and enhancing intestinal resistance in mice with DSS-induced enteritis. Reference 4 (Ran-Ressler RR, et al. Branched chain fatty acids reduce the incidence of necrotizing enterocolitis and alter gastrointestinal microbialecology in a neonatal rat model. PLoS One. 2011;6(12):e29032.) discloses that branched-chain fatty acids can reduce the risk of necrotizing enterocolitis (NEC) and can also increase the expression of IL-10 in the intestine.

[0007] Human milk oligosaccharides (HMOs) are the third most abundant solid component in breast milk, after lactose and fat, and are even more abundant than proteins. HMOs are a class of complex sugars with diverse structures. Based on their molecular structure, HMOs can be divided into two main categories: neutral fucosylated HMOs, neutral non-fucosylated HMOs, and acidic sialylated HMOs. Due to their unique structure, HMOs help infants establish a healthy gut microbiota, strengthen the immune system, and promote brain and cognitive development.

[0008] Previous studies have also explored the role of human milk oligosaccharides in intestinal inflammation. For example, reference 5 (CN121817480A) discloses a composition for improving neonatal necrotizing enterocolitis. This composition includes human milk oligosaccharides and Lactobacillus rhamnosus. It was found that the combination of human milk oligosaccharides and Lactobacillus rhamnosus can alleviate neonatal necrotizing enterocolitis by inhibiting the TLR4 / NF-κB inflammatory pathway, improving cellular oxidative stress, protecting the cell barrier, and reducing apoptosis. Summary of the Invention

[0009] The problem the invention aims to solve

[0010] Although existing technologies have conducted some research on the effects of branched-chain fatty acids and human milk oligosaccharides on intestinal function, they cannot be said to be sufficient or perfect. There is still a need to develop new nutritional compositions to improve intestinal function.

[0011] Given the various shortcomings of existing methods for intervening in intestinal inflammation, and to enrich the nutritional compositions available in the field, the present invention aims to develop a natural nutritional composition derived from milk that improves intestinal function.

[0012] Therefore, during the research on branched-chain fatty acids (BCFAs), this invention unexpectedly discovered that a specific combination of BCFAs has a certain efficacy in improving intestinal function, producing a synergistic effect. Further research revealed that the combination of this specific BCFAs with human milk oligosaccharides further enhances its efficacy in improving intestinal function. Therefore, this invention screens a nutritional composition that improves intestinal function.

[0013] Solution for solving the problem

[0014] To solve the above-mentioned technical problems, the present invention provides the following technical solution:

[0015] In a first aspect of the invention, a nutritional composition for improving intestinal function is provided, wherein the nutritional composition comprises isomer C15:0 (iso-C15:0) and transiso-C15:0 (anteiso-C15:0); the content of isomer C15:0 to the content of anteiso-C15:0 is in the ratio of 1:(0.1-10) by mass.

[0016] In some embodiments, the ratio of the content of iso-C15:0 to the content of anteiso-C15:0 is 1:(0.3-6), preferably 1:(0.5-3).

[0017] In some embodiments, the nutritional composition further comprises 3'-SL; preferably, the ratio of the sum of the contents of iso-C15:0 and anteiso-C15:0 to the contents of 3'-SL is 1:(0.01-20), more preferably 1:(0.05-15), and more preferably 1:(0.1-10).

[0018] In a second aspect of the invention, the use of the nutritional composition described in the first aspect of the invention in the preparation of products for improving intestinal function is provided.

[0019] In some implementations, the improvement of intestinal function includes at least one of non-therapeutic effects such as slowing the degree of intestinal damage, alleviating intestinal inflammation, and strengthening intestinal barrier function.

[0020] In some implementations, mitigating intestinal damage includes increasing intestinal length.

[0021] In some embodiments, the enhancement of intestinal barrier function includes promoting the expression of proteins related to intestinal tissue barrier integrity; optionally, the proteins related to intestinal tissue barrier integrity include at least one of ZO-1 protein, ZO-2 protein, Claudin-1, Claudin-2 and Claudin.

[0022] In a third aspect of the invention, the use of the nutritional composition described in the first aspect of the invention is provided in the preparation of products that help regulate intestinal immunity.

[0023] In some embodiments, the modulation of intestinal immunity includes at least one of inhibiting TNF-α production, inhibiting IL-6 production, and promoting IL-10 production.

[0024] The effects of the invention

[0025] In some embodiments, the branched-chain fatty acids (and optionally human milk oligosaccharides) in the nutritional composition provided by the present invention are natural food nutrients found in breast milk or cow's milk. They can be used for a long time, have good acceptance among people throughout their entire life cycle, and play a role in improving intestinal function.

[0026] In some embodiments, the nutritional composition containing branched-chain fatty acids iC15:0 and aC15:0 provided by the present invention has the effect of improving intestinal function and producing a synergistic effect. Specifically, it has the effects of reducing the degree of intestinal damage, balancing intestinal inflammation, and strengthening intestinal barrier function.

[0027] In some embodiments, the nutritional composition provided by the present invention, comprising branched-chain fatty acids iC15:0, aC15:0 and 3'-SL, synergistically improves intestinal function, specifically, it has the effects of reducing the degree of intestinal damage, balancing intestinal inflammation, and strengthening intestinal barrier function.

[0028] In some specific embodiments, the nutritional composition containing branched-chain fatty acids iC15:0 and aC15:0 provided by the present invention has the tendency to alleviate intestinal damage and promote the recovery of intestinal length to a normal developmental state. At the same time, both have the effect of regulating intestinal inflammation. In the range of 1:0.5-3, they have a synergistic effect of inhibiting the production of pro-inflammatory factors TNF-α and IL-6 and promoting the production of anti-inflammatory factor IL-10. They also strengthen intestinal barrier function and promote the mRNA expression of intestinal tight junction proteins Claudin-1, Claudin-2 and Claudin, as well as ZO-1 and ZO-2 proteins. When these two branched-chain fatty acids are compounded with 3'-SL in a ratio of 1:0.1-10, the resulting complex further enhances its efficacy in alleviating intestinal damage, balancing intestinal inflammation, and strengthening intestinal barrier function. Attached Figure Description

[0029] Figure 1 The change in body weight of mice in each group after 14 days of intervention.

[0030] Figure 2 Comparison of intestinal length among different groups of mice. Detailed Implementation

[0031] Various exemplary embodiments, features, and aspects of the present invention will be described in detail below. The term "exemplary" as used herein means "serving as an example, embodiment, or illustration." Any embodiment described herein as "exemplary" is not necessarily to be construed as superior to or better than other embodiments.

[0032] Furthermore, to better illustrate the present invention, numerous specific details are set forth in the following detailed embodiments. Those skilled in the art should understand that the present invention can be practiced without certain specific details. In other instances, methods, means, apparatus, and steps well known to those skilled in the art have not been described in detail in order to highlight the spirit of the present invention.

[0033] Unless otherwise stated, all units used in this specification are international standard units, and all numerical values ​​and ranges appearing in this invention should be understood to include systematic errors that are unavoidable in industrial production.

[0034] In this specification, the word "may" has two meanings: to perform a certain process and not to perform a certain process.

[0035] In this specification, references to "some specific / preferred embodiments," "other specific / preferred embodiments," "implementation," etc., refer to specific elements (e.g., features, structures, properties, and / or characteristics) related to that embodiment, which are included in at least one of the embodiments described herein and may or may not be present in other embodiments. Furthermore, it should be understood that these elements may be combined in any suitable manner in various embodiments.

[0036] In this specification, "optional" and "optionally" mean that the events or circumstances described below may or may not occur, and the description includes both cases where the events or circumstances occur and cases where the events or circumstances do not occur.

[0037] In this specification, the range of values ​​referred to as "value A to value B" refers to the range including the endpoint values ​​A and B.

[0038] In this invention, the word "may" has two meanings: to perform a certain process and not to perform a certain process.

[0039] In this invention, the terms “a”, “an”, or “the” can mean “one”, “one or more”, “at least one”, or “one or more”.

[0040] In this invention, the terms "comprising," "having," "including," or "containing" can mean included or open-ended, and do not exclude additional, uncited elements or method steps. At the same time, "comprising," "having," "including," or "containing" can also mean closed-ended, excluding additional, uncited elements or method steps.

[0041] In this invention, the term "about" is used to define that the numerical ranges and parameters of this invention are approximate values, while specific related values ​​have been presented as precisely as possible. Unless otherwise explicitly stated, it should be understood that all ranges, quantities, values, and percentages used in this invention are modified by "about". Here, "about" generally means that the actual value is within ±5%, ±3%, ±1%, or ±0.5% of a specific value or range.

[0042] Unless otherwise defined, other technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0043] I. Uses of nutritional compositions to improve intestinal function

[0044] This invention has unexpectedly discovered that nutritional compositions containing iso-C15:0 (also referred to as iC15:0) and anteiso-C15:0 (also referred to as aC15:0) have the effect of improving intestinal function. Furthermore, when iso-C15:0 and anteiso-C15:0 are combined, they have a synergistic effect. Further, when iso-C15:0 and anteiso-C15:0 are combined with 3'-SL, a further synergistic effect is produced, which can further enhance the efficacy of improving intestinal function. Therefore, the nutritional compositions containing iso-C15:0 and anteiso-C15:0, or containing iso-C15:0, anteiso-C15:0, and 3'-SL, as well as products or compositions that have added to or used said nutritional compositions, all have the effect of improving intestinal function. The improvement of intestinal function described in this invention is not intended for the prevention or treatment of diseases.

[0045] Furthermore, the present invention provides the use of a nutritional composition containing iso-C15:0 and anteiso-C15:0 or containing iso-C15:0, anteiso-C15:0 and 3'-SL in the preparation of products or compositions that improve intestinal function.

[0046] In some embodiments, the improvement of gut function includes non-therapeutic relief of gut inflammation, which is not of a medically pathological degree. In some embodiments, the improvement of gut function includes non-therapeutic mitigation of gut damage, which is not of a medically pathological degree.

[0047] In some specific implementations, mitigating intestinal damage includes increasing intestinal length.

[0048] It should be noted that the intestinal inflammation and intestinal damage mentioned in this instruction manual refer to a non-pathological state in which the intestines are not fully healthy due to inflammation or other factors.

[0049] In some embodiments, the improvement of intestinal function includes strengthening intestinal barrier function for non-therapeutic purposes. In some specific embodiments, strengthening intestinal barrier function includes promoting the expression of proteins related to intestinal tissue barrier integrity. In some optional embodiments, the proteins related to intestinal tissue barrier integrity include at least one of ZO-1 protein, ZO-2 protein, Claudin-1, Claudin-2, and Claudin.

[0050] II. Uses that help regulate intestinal immunity

[0051] This invention unexpectedly discovered that nutritional compositions containing iso-C15:0 and anteiso-C15:0 have a modulating effect on intestinal immunity. Furthermore, when iso-C15:0 and anteiso-C15:0 are combined, they exhibit a synergistic effect. Moreover, when iso-C15:0 and anteiso-C15:0 are combined with 3'-SL, a further synergistic effect is produced, which can further enhance the efficacy of intestinal immune modulation. Therefore, the nutritional compositions containing iso-C15:0 and anteiso-C15:0, or containing iso-C15:0, anteiso-C15:0, and 3'-SL, as well as products or compositions that have added to or used said nutritional compositions, all have a modulating effect on intestinal immunity. The intestinal immune modulation described in this invention is not intended for the prevention or treatment of diseases.

[0052] Furthermore, the present invention provides the use of nutritional compositions containing iso-C15:0 and anteiso-C15:0 or containing iso-C15:0, anteiso-C15:0 and 3'-SL in the preparation of products or compositions that help regulate intestinal immunity.

[0053] In some embodiments, the modulation of intestinal immunity includes at least one of inhibiting TNF-α production, inhibiting IL-6 production, and promoting IL-10 production.

[0054] III. Nutritional Composition

[0055] In the above aspects of the present invention, the nutritional composition of the present invention comprises isomer C15:0 (iso-C15:0) and anti-isomer C15:0 (anteiso-C15:0).

[0056] In this invention, "iso-branched fatty acid" or "iso-branched fatty acid (iso-BCFA)" refers to a fatty acid with a methyl branch on the penultimate carbon atom of the fatty acid molecule's carbon chain backbone. "Anti-iso-branched fatty acid" or "antiso-BCFA" refers to a fatty acid with a methyl branch on the penultimate carbon atom of the fatty acid molecule's carbon chain backbone.

[0057] Furthermore, regarding the content ratio of iso-C15:0 and anteiso-C15:0 in the nutritional composition, it has been found that the technical effects of the present invention can be obtained by setting the following mass ratio: the content ratio of iso-C15:0 to anteiso-C15:0 is 1:(0.1-10).

[0058] In some preferred embodiments, the content of iso-C15:0 is in the ratio of the content of anteiso-C15:0 to that of 1:(0.2-8).

[0059] In some more preferred embodiments, the ratio of the iso-C15:0 content to the anteiso-C15:0 content is 1:(0.3-6), 1:(0.4-5), or 1:(0.4-4).

[0060] In some further preferred embodiments, the content of iso-C15:0 is in the ratio of the content of anteiso-C15:0 to 1:(0.5-3).

[0061] For example, the ratio of the content of iso-C15:0 to the content of anteiso-C15:0 is 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:1.1, 1:1.3, 1:1.5, 1:1.7, 1:2, 1:2.1, 1:2.3, 1:2.5, 1:2.7, 1:2.9, or 1:3.

[0062] In some further preferred embodiments, the ratio of the iso-C15:0 content to the anteiso-C15:0 content is 1:(0.8-5), 1:(1-5), 1:(1-4), or 1:(1.2-3.5).

[0063] In some further preferred embodiments, the ratio of the iso-C15:0 content to the anteiso-C15:0 content is 1:(1.8-5), 1:(2-5), 1:(2-4), or 1:(2.2-3.5).

[0064] In addition to the iso-C15:0 and anteiso-C15:0 fatty acids described above, the nutritional composition of the present invention may optionally contain other types of fatty acids, or may not contain them. These other types of fatty acids may be branched or linear. Furthermore, there is no particular limitation on the amount of these other types of fatty acids used in principle.

[0065] This invention does not particularly limit the source of branched-chain fatty acids. For example, they can be isolated and extracted from natural substances containing branched-chain fatty acids, such as bacterial biofilms, animal sebum, mammary tissue, and plants. They can also be prepared through biological or chemical synthesis, such as using microorganisms or *C. elegans* for synthesis. Exemplarily, branched-chain fatty acids are synthesized in microorganisms or *C. elegans* using branched-chain amino acids as substrates through enzymatic reactions. Currently, the commonly used method in this field is to isolate and extract branched-chain fatty acids from lanolin. Lanolin undergoes a saponification reaction to prepare free lanolin alcohol and free lanolin acid soap. Branched-chain fatty acids from lanolin are then obtained through multiple steps, including alcohol-soap separation and fatty acid preparation and extraction.

[0066] In some embodiments, the nutritional composition of the present invention further comprises human milk oligosaccharides.

[0067] Human milk oligosaccharides (HMOs) are a collective term for oligosaccharides with a degree of polymerization ≥3 that are naturally found in human milk. They are formed by modifying the terminal positions of lactose molecules with five monomers: glucose (Glc), galactose (Gal), N-acetylglucosamine (GlcNAc), fucose (Fuc), and N-acetylneuraminic acid (Neu5Ac). Each HMO molecule contains 3 to 32 monosaccharides linked by different glycosidic bonds, contributing to the diversity and complexity of HMOs.

[0068] HMOs are mainly composed of five core monomers: glucose, sialic acid, fucose, N-acetylglucosamine, and galactose. Different HMOs exhibit different fucosylation and sialylation, thus HMOs in breast milk can be classified into neutral fucosylated HMOs, acidic sialylated HMOs, and neutral non-fucosylated HMOs.

[0069] Currently, the main methods for producing human milk oligosaccharides include chemical synthesis, enzymatic synthesis, and bioengineering synthesis. At the same time, these components can also be indirectly introduced from various existing milk components.

[0070] It should be noted that the use of the various human milk oligosaccharides described above in this invention shall comply with the requirements of local laws and regulations. In some cases, where permitted by laws and regulations, these components may be directly introduced into the composition or product as individual raw materials; in other cases, where permitted by laws and regulations, they may be indirectly introduced into the composition or product through the addition of qualified milk raw materials.

[0071] This invention does not specifically limit the source of human milk oligosaccharides; typically, they can be derived from various milk-containing raw materials, such as animal milk raw materials (e.g., cow's milk, milk powder, etc.). In this invention, "animal milk" is used to refer to the liquid obtained from the mammary glands of mammals in lactation.

[0072] The term “animal milk” should be interpreted broadly and encompass both raw milk (i.e., the liquid obtained directly from the mammary glands) and standardized dairy products.

[0073] The human milk oligosaccharides of the present invention can exist in a liquid state, or in a semi-solid or solid state.

[0074] In some preferred embodiments, the human milk oligosaccharide comprises 3'-sialylated lactose (3'-SL).

[0075] In some embodiments, the ratio of the sum of the contents of iso-C15:0 and anteiso-C15:0 to the content of 3'-SL by weight is 1:(0.01-20), preferably 1:(0.05-15), more preferably 1:(0.1-10).

[0076] In some preferred embodiments, the ratio of the sum of the contents of iso-C15:0 and anteiso-C15:0 to the content of 3'-SL is 1:(0.03-18), 1:(0.05-15), or 1:(0.08-12).

[0077] In some more preferred embodiments, the sum of the contents of iso-C15:0 and anteiso-C15:0 is given by a ratio of 1:(0.1-10) to the content of 3'-SL.

[0078] For example, the ratio of the sum of the contents of iso-C15:0 and anteiso-C15:0 to the content of 3'-SL is 1:0.1, 1:0.3, 1:0.5, 1:0.7, 1:0.9, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10.

[0079] In some further preferred embodiments, the ratio of the sum of the contents of iso-C15:0 and anteiso-C15:0 to the content of 3'-SL is 1:(2-8), 1:(3-7), or 1:(4-6).

[0080] Product or composition

[0081] The products or compositions of this invention contain or use the above-mentioned nutrient compounds, thereby including iso-C15:0 and anteiso-C15:0 in the products or compositions of this invention. In some optional embodiments, the products or compositions of this invention also contain human milk oligosaccharides, such as 3'-SL.

[0082] This invention does not impose any particular limitation on the absolute content of iso-C15:0, anteiso-C15:0, and optional 3'-SL in the product, as long as they comply with relevant laws and regulations, such as complying with the quantitative limits of fatty acids in food as specified in GB 5009.168-2016 "National Food Safety Standard - Determination of Fatty Acids in Food". In some embodiments, the content of C15:0 is not higher than 0.0033 g / 100g in solid products, for example, 0.0001~0.0033 g / 100g; and not higher than 0.0013 g / 100g in liquid products, for example, 0.0001~0.0013 g / 100g.

[0083] In some implementations, the product includes food and / or health food.

[0084] This invention does not specifically limit the types of food products, but may include candies, beverages, dairy products, or baked goods. Examples of candies include hard candies, shortbread candies, gel candies, compressed candies, and aerated candies. Examples of beverages include carbonated drinks, tea drinks, coffee drinks, fruit and vegetable juices, and lactic acid bacteria drinks. Examples of dairy products include fermented milk, cheese, and milk powder. Examples of baked goods include bread, cakes, and biscuits. Furthermore, the food products described in this invention can also be health foods, such as various types of oral preparations, including tablets, pills, granules, powders, capsules, and oral liquids.

[0085] In addition to iso-C15:0 and anteiso-C15:0 and optional 3'-SL in the nutritional composition, in some embodiments, depending on the type of product and the end needs of the target audience, the product of the present invention may also contain any one or more of the following ingredients: plant-based ingredients, animal dairy ingredients, animal meat ingredients, functional additives, and any acceptable excipients.

[0086] Examples of plant-based ingredients include fruits such as figs, pomegranates, kiwis, oranges, tangerines, pineapples, strawberries, apples, bananas, grapes, pears, cherries, blueberries, blackberries, blackcurrants, cranberries, raspberries, melons, amla, and bilberries, or their extracts; vegetables such as onions, cucumbers, tomatoes, cauliflower, carrots, spinach, kale, Brussels sprouts, garlic, basil, and oregano, or their extracts; grains such as rice (indica, japonica, glutinous rice), cereals (wheat, barley, oats, rye), corn, sorghum, millet, sorghum, yellow millet, buckwheat, soybeans, broad beans, peas, mung beans, red beans, and kidney beans, or their extracts; nuts such as walnuts, pistachios, cashews, hazelnuts, almonds, apricot kernels, pine nuts, peanuts, sunflower seeds, chestnuts, macadamia nuts, and ginkgo nuts, or their extracts; coffee or its extracts; and some medicinal and edible herbal medicines or their extracts.

[0087] Animal dairy product ingredients can include fresh milk from mammals such as cows, sheep, and camels, as well as reprocessed dairy products such as whole milk powder, skim milk powder, whey protein concentrate, desalted whey powder, whey protein powder, and hydrolyzed whey protein powder.

[0088] Examples of animal meat product ingredients include those from pork, beef, mutton, seafood, or poultry.

[0089] Examples of functional additives include vitamin supplements, mineral supplements, nucleotide supplements, dietary fiber, and functional polyunsaturated fatty acid supplements.

[0090] Any acceptable excipients may include solvents, antioxidants, antibacterial agents, thickeners, diluents, cosolvents, stabilizers, emulsifiers, fillers, disintegrants, lubricants, coating materials, anti-caking agents, flavoring agents, sweeteners, food flavorings, and food colorings.

[0091] Example

[0092] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0093] Experimental methods

[0094] 1. Model building and nutritional intervention

[0095] Six-week-old male C57BL / 6 mice were selected and fed a standard diet freely for one week for acclimatization observation. They were randomly divided into three groups: a control group, an LPS model group (hereinafter referred to as the model group), and an LPS model group plus different nutrient intervention groups, for a total of 12 groups, with 6 mice in each group. After one week of acclimatization, mice in the LPS model group and the LPS model group plus different nutrient intervention groups received intraperitoneal injections of 1.0 mg / kg LPS, while the control group received an equal volume of physiological saline, for 7 consecutive days. After modeling was completed, mice in the control group and the LPS model group were administered 0.2 mL of sterile PBS by gavage for 14 consecutive days, while mice in the intervention groups were administered 0.2 mL of different nutrient mixtures by gavage for 14 consecutive days. Half an hour after the last gavage, the mice were enucleated to collect blood, euthanized by cervical dislocation, and all tissues were collected for later use.

[0096] 2. Weight measurement

[0097] After the modeling was completed, the weight changes of the mice were recorded weekly.

[0098] 3. Mouse intestinal assay method

[0099] After euthanizing the mice, dissected them to remove intestinal tissue, which was then laid flat on an ice box and stretched taut with zero tension. A ruler was placed close to the intestinal tract, and values ​​were read along the central axis of the intestinal tract. Each segment was measured three times, and the arithmetic mean was taken as the final data for that sample.

[0100] 3. Assessment of changes in intestinal inflammation in mice

[0101] The inflammatory factors (IL-6, TNF-α, IL-10) in the intestine and serum were measured using an ELISA kit.

[0102] 4. Assessment of changes in the intestinal barrier in mice

[0103] The expression of ZO-1, ZO-2, claudin-1, claudin-2 and occludin was detected by RT-qPCR to assess the integrity of the intestinal barrier.

[0104]

[0105] Examples and Comparative Examples

[0106] Experimental intervention design

[0107] In this invention, the effects of branched-chain fatty acids iC15:0 and aC15:0 alone and in combination on intestinal inflammation and intestinal barrier function were first investigated. Specifically, Comparative Examples 1 and 2 showed that the gavage concentration of the two groups of mice was 0.2 mg / mL. Examples 1-3 showed that the ratio of iC15:0 to aC15:0 was varied when the total solid gavage concentration of the three groups of mice was 0.2 mg / mL, and the ratios were 1:0.5, 1:1.5 and 1:3, respectively. The effects of combining two branched-chain fatty acids were investigated using Comparative Examples 1-2 and Examples 1-3. The effects of combining the two branched-chain fatty acids with human milk oligosaccharide 3'-SL were then examined. Comparative Example 3 involved 3'-SL alone at a concentration of 2.2 mg / mL. Examples 4-6 were intervention groups with total branched-chain fatty acid to 3'-SL ratios of 1:0.1, 1:1, 1:5, and 1:10, respectively. In these three examples, the total amount of solid branched-chain fatty acids was 0.2 mg / mL, but the ratios of iC15:0 and aC15:0 were 1:0.5, 1:1.5, and 1:3, respectively. The specific nutrient intervention concentrations for different groups are shown in Table 2.

[0108]

[0109] Test case

[0110] Test Example 1: Evaluation of Basal Growth Indicators in Mice

[0111] The changes in body weight of mice in the control group, model group, and 8 intervention groups after 14 days of intervention are as follows: Figure 1 As shown in the figure, the model group mice showed a significant decrease in body weight compared to the control group mice after 14 days of intervention. However, after intervention with branched-chain fatty acids iC15:0, aC15:0, and human milk oligosaccharide 3'-SL monomers or combinations, the body weight showed varying degrees of recovery towards the normal control group. This preliminarily indicates that several nutrients have a certain effect on improving the body weight of mice.

[0112] Test Example 2: Effect of nutritional intervention on intestinal length in mice with enteritis

[0113] This invention investigated the intestinal length of mice in different groups after LPS gavage. The experimental results are as follows: Figure 2 As shown in the figure, compared with the control group of normal mice, the intestinal length of the model group mice after LPS modeling was shortened, indicating that the intestine was severely damaged. However, it can also be seen that after intervention with two branched-chain fatty acids iC15:0 and aC15:0 alone and in combination, and intervention with human milk oligosaccharide 3'-SL alone and intervention with 3'-SL in combination with the two branched-chain fatty acids, the intestinal length showed a tendency to recover to the control group to varying degrees. This preliminarily suggests that several nutrient monomers and different combinations have a certain degree of inhibitory or therapeutic effect on intestinal damage.

[0114] Test Example 3: The Regulatory Effect of Nutritional Intervention on Intestinal Inflammation in Enteritis Mice

[0115] This invention used animal experiments to discover and verify the ameliorative effects of branched-chain fatty acids and their compositions on intestinal inflammation, as shown in Tables 3 and 4-1 to 4-3. TNF-α and IL-6 are key pro-inflammatory factors, and their elevated levels are core markers driving neuroinflammatory responses and exacerbating neuronal damage. IL-10 is an important anti-inflammatory factor that can inhibit the inflammatory process and promote tissue repair.

[0116] Table 3 shows the levels of two pro-inflammatory factors, TNF-α and IL-6, and the anti-inflammatory factor IL-10 in the intestinal tissues of mice in the control group, model group, and different nutritional intervention groups. Tables 4-1 to 4-3 show the results of the statistical significance analysis of the differences among the various inflammatory factor groups. TNF-α is a core initiator of the inflammatory cascade, potently pro-inflammatory and inducing apoptosis, playing a key role in autoimmune diseases, sepsis, and metabolic diseases. Table 3 shows that the TNF-a level in the intestinal tissue of the control group mice was 26.31 pg / mL. After LPS modeling, the TNF-a level in the brain tissue of mice significantly increased to 128.26 pg / mL, which was significantly higher in the model group than in the control group (p < 0.0001). Comparative Examples 1 and 2 showed that treatment with 0.2 mg / mL iC15:0 and aC15:0, respectively, reduced the TNF-a level in the model mice to 105.32 pg / mL and 95.82 pg / mL, which were also significantly lower than in the model group (p < 0.0001). This indicates that the levels of TNF-a, whether alone or in combination, are significantly lower in the model group. The pre-model mice showed a reduction in the level of the pro-inflammatory factor TNF-α in intestinal tissue. When two branched-chain fatty acids were combined and the total concentration was fixed at 0.2 mg / mL, the concentration ratio of the two was adjusted to 1:0.5 (Example 1), 1:1.5 (Example 2), and 1:3 (Example 3). The level of the pro-inflammatory factor TNF-α was further reduced to 78-83 pg / mL. The difference significance analysis showed that the levels in Examples 1-3 were significantly lower than those in the model group, and significantly lower than those in Comparative Examples 1 and 2. This indicates that when iC15:0 and aC15:0 are in the range of 1:0.5-3, they have a synergistic effect in reducing the level of the pro-inflammatory factor TNF-α. We further investigated the synergistic effects of human milk oligosaccharide 3'-SL and two branched-chain fatty acids. Comparative Example 3 showed that after intervention with 3'-SL alone at a concentration of 2.2 mg / mL, the amount of TNF-a in the intestinal tissue of mice was 98.15 pg / mL, which was significantly lower than that in the model group (p < 0.0001). When the total concentration of 2.2 mg / mL was fixed, the ratio of total iC15:0 and aC15:0 to 3'-SL was 1:0.1 (Example 4), 1:1 (Example 5), 1:5 (Example 6), and 1:10 (Example 7), respectively, the amount of TNF-a was further reduced to 54-75 pg / mL. After significance analysis, it was found that Examples 4-7 were significantly lower than the model group, significantly lower than Comparative Example 3 with 3'-SL alone, and significantly lower than Examples 1-3 with two branched-chain fatty acids in different proportions (as shown in Tables 4-1 to 4-3). This indicates that the combination of 3'-SL further reduced the content of TNF-a.

[0117] IL-6 is a core molecule in the inflammatory cascade, participating in acute-phase inflammatory responses and metabolic regulation. In this invention, the IL-6 level in the intestinal tissue of control mice was 38.55 pg / mL. After LPS modeling, the IL-6 level in the mouse intestinal tissue significantly increased to 88.67 pg / mL, significantly higher than the control group (p < 0.0001). Comparative Examples 1 and 2 showed that treatment with 0.2 mg / mL iC15:0 and aC15:0, respectively, reduced the IL-6 levels in model mice to 75.04 pg / mL and 72.96 pg / mL, also significantly lower than the model group (p < 0.0001). This indicates that the levels of IL-6 in the model are significantly lower when either of the two branched-chain fatty acids is used alone. The study showed that the levels of pro-inflammatory IL-6 in the intestinal tissue of mice with enteritis could be reduced. When two branched-chain fatty acids were combined and the total concentration was fixed at 0.2 mg / mL, the concentration ratios of the two fatty acids were adjusted to 1:0.5, 1:1.5, and 1:3 (Examples 1-3). The levels of pro-inflammatory IL-6 were further reduced to 62-66 pg / mL. The difference significance analysis showed that the levels in Examples 1-3 were significantly lower than those in the model group, and significantly lower than those in Comparative Examples 1 and 2. This indicates that when iC15:0 and aC15:0 are in the range of 1:0.5-3, they have a synergistic effect in reducing the level of pro-inflammatory IL-6. We further investigated the synergistic effects of 3'-SL and the two branched-chain fatty acids. Comparative Example 3 showed that the amount of IL-6 in the intestinal tissue of mice after administration of 3'-SL alone was 75.41 pg / mL, which was significantly lower than that in the model group (p < 0.0001). When the total concentration was fixed at 2.2 mg / mL, and the ratio of total iC15:0 and aC15:0 to 3'-SL was 1:0.1 (Example 4), 1:1 (Example 5), 1:5 (Example 6), and 1:10 (Example 7), the amount of IL-6 was further reduced to the range of 54-57 pg / mL. After significance analysis, it was found that Examples 4-7 were significantly lower than the model group, significantly lower than Comparative Example 3 with 3'-SL alone, and significantly lower than Examples 1-3 with the two branched-chain fatty acids combined in different proportions (as shown in Tables 4-1 to 4-3). This indicates that the combination of 3'-SL further reduced the content of IL-6.

[0118] The anti-inflammatory cytokine IL-10 is mainly secreted by regulatory T cells (Tregs), macrophages, B cells, and dendritic cells. It plays a role in inhibiting excessive immune responses and maintaining immune homeostasis. In this invention, it was found that the amount of IL-10 in the intestinal tissue of model mice was significantly lower than that in the control group. However, treatment with iC15:0 and aC15:0 at a concentration of 0.2 mg / mL significantly increased the content of anti-inflammatory IL-10 in model mice (p < 0.0001). This indicates that intervention with the two branched-chain fatty acids alone can increase the amount of anti-inflammatory IL-10 in the intestinal tissue of model mice. When the model mice were given a nutritional composition of the two branched-chain fatty acids (Examples 1-3), the content of anti-inflammatory IL-10 was further increased. The difference significance analysis showed that the levels in Examples 1-3 were significantly higher than those in the model group, and significantly higher than those in Comparative Examples 1 and 2. This indicates that when iC15:0 and aC15:0 are in the range of 1:0.5-3, they have a synergistic effect in increasing the amount of anti-inflammatory IL-10. Furthermore, we investigated the synergistic effects of 3'-SL and the two branched-chain fatty acids. Comparative Example 3 showed that the amount of IL-10 in the intestinal tissue of mice after administration of 3'-SL alone was significantly higher than that in the model group (p < 0.0001). When the total concentration was fixed, the ratio of the total amount of iC15:0 and aC15:0 to 3'-SL was 1:0.1, 1:1, 1:5, and 1:10 (Examples 4-7), respectively, the amount of IL-10 was further increased. After significance analysis, it was found that Examples 4-7 were significantly higher than the model group, significantly higher than Comparative Example 3, and significantly higher than Examples 1-3, which combined the two branched-chain fatty acids in different proportions (as shown in Tables 4-1 to 4-3). This indicates that the combination of 3'-SL further increased the level of the anti-inflammatory factor IL-10.

[0119] The above experimental results indicate that when the two branched-chain fatty acids iC15:0 and aC15:0 are combined in a ratio of 1:0.5-3, the resulting nutritional composition can synergistically reduce the levels of pro-inflammatory factors TNF-α and IL-6 in intestinal tissue and increase the level of anti-inflammatory factor IL-10. Furthermore, when 3'-SL is added, it is found that when the total amount of the two branched-chain fatty acids iC15:0 and aC15:0 and the ratio of 3'-SL are 1:0.1-10, the two have a synergistic effect in regulating immune factors.

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127] Test Example 4: The Regulatory Effect of Nutritional Intervention on Intestinal Barrier Function in Enteritis Mice

[0128] Next, we used qRT-PCR to analyze the expression of proteins related to intestinal barrier integrity to reflect the effects of LPS modeling and different nutrient / combination interventions on intestinal barrier function. It is well known that intestinal barrier integrity is closely related to the occurrence and development of various diseases, and tight junctions between epithelial cells are an important component of the intestinal mechanical barrier. These junctions are mainly composed of molecules such as ocludin, claudin, mucin (MUC), and zonula occludens (ZO, also known as tight junction proteins, mainly including ZO-1, ZO-2, and ZO-3). In this invention, we mainly examined the expression levels of two major ZO proteins and three Claudin proteins mRNA (as shown in Table 5). Tables 6 and 7 show the results of the significant differences between the two types of tight junction protein groups.

[0129] ZO-1 and ZO-2 are cytoplasmic scaffold proteins. As "bridges" connecting transmembrane proteins to the cytoskeleton, they are responsible for maintaining the integrity and stability of tight junction structures. Table 5 shows that the mRNA expression levels of ZO-1 and ZO-2 proteins in the intestinal tissue of the model group mice were significantly lower than those in the control group. Treatment with 0.2 mg / mL iC15:0 and aC15:0 in the model mice increased the mRNA expression levels of these two tight junction proteins to varying degrees. However, after differential significance analysis, there was no significant difference in the mRNA expression levels of ZO-1 protein between iC15:0 and the model group (p > 0.05), while there was a significant difference in the mRNA expression levels of ZO-1 protein between aC15:0 and the model group (p < 0.05). In contrast, the mRNA expression levels of ZO-2 protein in the intestinal tissue of mice in both branched-chain fatty acid intervention groups were significantly higher. In the model group, this indicates that the individual intervention of the two branched-chain fatty acids in the model mice had a limited effect on increasing the mRNA expression levels of ZO-1 and ZO-2 proteins in the intestinal tissue. However, when the model mice were given a nutritional composition of the two branched-chain fatty acids (Examples 1-3), it was found that the mRNA expression levels of both proteins were significantly increased. Moreover, the difference significance analysis showed that the mRNA expression levels of ZO-1 and ZO-2 in Examples 1-3 were significantly higher than those in the model group, and significantly higher than those in Comparative Examples 1 and 2 (as shown in Table 6). This indicates that when iC15:0 and aC15:0 are in the range of 1:0.5-3, they have a synergistic effect on increasing the mRNA expression levels of ZO-1 and ZO-2 proteins. We further investigated the synergistic effects of 3'-SL and the two branched-chain fatty acids. Comparative Example 3 showed that the mRNA expression levels of ZO-1 and ZO-2 proteins in the intestinal tissue of mice after administration of 3'-SL alone were significantly higher than those in the model group (p < 0.0001). When the total concentration was fixed and the ratio of total iC15:0 and aC15:0 to 3'-SL was 1:0.1, 1:1, 1:5, and 1:10 (Examples 4-7), the mRNA expression levels of ZO-1 and ZO-2 proteins were further increased. After significance analysis, Examples 4-7 were significantly higher than the model group, significantly higher than Comparative Example 3, and significantly higher than Examples 1-3 with different ratios of the two branched-chain fatty acids (as shown in Table 6). This indicates that the combination of 3'-SL further enhances the mRNA expression levels of ZO-1 and ZO-2 proteins.

[0130] Claudin-1 is a barrier-forming protein. It is a major "sealing protein" that makes up tight junction strands, dominating the sealing of the epithelial barrier by controlling the permeation of ions and small molecules through the paracellular pathway. Claudin-2 is a pore-forming protein. It forms specific cation and water channels in tight junctions, and its upregulation is generally associated with weakened barrier function and increased "leakage." Occludin is a barrier function regulatory protein. Embedded in tight junctions, it is not structurally essential, but it is crucial for the dynamic regulation of barrier function and the maintenance of "gate" function. As shown in Table 5, the mRNA expression level of Claudin-1 protein in the intestinal tissue of the model group mice was reduced to about 1 / 4 of that in the control group, and the mRNA expression levels of Claudin-2 and Claudin proteins were reduced to about 1 / 5 of those in the control group. After treatment with 0.2 mg / mL iC15:0 and aC15:0, except that the mRNA expression level of Claudin-1 protein in the intestinal tissue of the aC15:0 intervention group was not different from that in the model group, the mRNA expression levels of the three Claudin proteins in iC15:0 and the mRNA expression levels of Claudin and Claudin-2 proteins in aC15:0 were significantly higher than those in the model group. This also indicates that the intervention of iC15:0 and aC15:0 alone has limited effect on increasing the mRNA expression level of Claudin protein in the intestinal barrier. When the model mice were given a nutritional composition consisting of two branched-chain fatty acids (Examples 1-3), the mRNA expression levels of the three proteins were further increased. Furthermore, a significant difference analysis revealed that the mRNA expression levels of Claudin-1, Claudin-2, and Claudin proteins in Examples 1-3 were significantly higher than those in the model group, and significantly higher than those in Comparative Examples 1 and 2 (as shown in Table 7). This indicates that when iC15:0 and aC15:0 are within the range of 1:0.5-3, they synergistically enhance the mRNA expression levels of Claudin-1, Claudin-2, and Claudin proteins.We further investigated the synergistic effects of 3'-SL and the two branched-chain fatty acids. Comparative Example 3 showed that the mRNA expression levels of Claudin-1, Claudin-2, and Claudin proteins in the intestinal tissue of mice after administration of 3'-SL alone were significantly higher than those in the model group (p < 0.0001). When the total concentration was fixed and the ratio of total iC15:0 and aC15:0 to 3'-SL was 1:0.1, 1:1, 1:5, and 1:10 (Examples 4-7), the mRNA expression levels of Claudin-1, Claudin-2, and Claudin proteins were further increased. After significance analysis, Examples 4-7 were significantly higher than the model group, significantly higher than Comparative Example 3, and significantly higher than Examples 1-3, which combined the two branched-chain fatty acids in different proportions (as shown in Table 7). This indicates that the combination of 3'-SL further enhances the mRNA expression levels of Claudin-1, Claudin-2, and Claudin proteins.

[0131]

[0132]

[0133]

[0134]

[0135]

[0136] It should be noted that although the technical solution of the present invention has been described with specific examples, those skilled in the art will understand that the present invention should not be limited thereto.

[0137] The various embodiments of the present invention have been described above. These descriptions are exemplary and not exhaustive, nor are they limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is chosen to best explain the principles, practical applications, or technological improvements to the embodiments in the market, or to enable others skilled in the art to understand the embodiments disclosed herein.

Claims

1. A nutritional composition for improving gut function, wherein, The nutritional composition comprises iso-C15:0 and anteiso-C15:0, wherein the content of iso-C15:0 to the content of anteiso-C15:0 is 1:(0.1-10) by mass. The nutritional composition further comprises 3'-SL, wherein the sum of the contents of iso-C15:0 and anteiso-C15:0 by mass is in the ratio of the contents of 3'-SL to 1:(0.01-20).

2. Use of the nutritional composition of claim 1 in the preparation of products for improving intestinal function.

3. Use according to claim 2, wherein, The improvement of intestinal function includes at least one of non-therapeutic measures such as slowing the degree of intestinal damage, relieving intestinal inflammation, and strengthening intestinal barrier function.

4. Use according to claim 3, wherein, The reduction of intestinal damage includes increasing intestinal length.

5. Use according to claim 3, wherein, The enhancement of intestinal barrier function includes promoting the expression of proteins related to intestinal tissue barrier integrity.

6. Use according to claim 5, wherein, The intestinal tissue barrier integrity-related proteins include at least one of ZO-1 protein, ZO-2 protein, Claudin-1, Claudin-2, and Claudin.

7. Use of the nutritional composition of claim 1 in the preparation of products that help regulate intestinal immunity.

8. The use according to claim 7, wherein, The regulation of intestinal immunity includes at least one of inhibiting TNF-α production, inhibiting IL-6 production, and promoting IL-10 production.

Images