Composition and food composition for improving muscular function

Abstract

The present invention relates to a composition and a health functional food for improving muscular function, each comprising sialyllactose as an active ingredient to reduce blood lactate levels and increase the expression of genes related to muscle function improvement, thereby allowing for the improvement of muscular functions such as anti-fatigue properties of muscles, muscular endurance, exercise performance, and the like without excessively increasing the concentration of lactic acid, which is a fatigue-inducing substance.

Description

Composition and food composition for improving muscle function

[0001] The present invention relates to a composition and a food composition for improving muscle function containing sialyl oligosaccharide.

[0002]

[0003] Excessive exercise increases the blood concentration of lactate, which is known to be the most important substance that causes exercise fatigue by interfering with muscle contraction and is produced during anaerobic exercise (Karlsson J et al., 1970, Kim et al. 1997).

[0004] When a lot of energy is needed in a short period of time due to excessive exercise, glucose is converted to lactic acid. Lactic acid is produced in muscle cells, red blood cells, brain, gastrointestinal tract, skin, etc., but under normal conditions, it exists in the blood at low concentrations.

[0005] The normal energy production process uses glucose and oxygen in the mitochondria to produce ATP. However, when oxygen becomes insufficient due to excessive exercise or other reasons, the process switches to anaerobic energy production. Pyruvate is rapidly converted to lactate by lactate dehydrogenase (LDH). This is a series of reactions that, although less efficient, produces at least some ATP. The lactate produced in this way is excreted into the blood, where it is converted to pyruvate in the liver, which then leads to glucose synthesis and maintains the blood lactate concentration.

[0006] In general, the rate of lactate removal from the liver is lower than the rate of lactate production in the muscles, or the blood lactate concentration may temporarily increase rapidly due to excessive exercise, and the increase in blood lactate levels during exercise is known to be proportional to the exercise intensity (Lagally KM, 2002).

[0007] Therefore, the ability to remove blood lactate during exercise or recovery is correlated with exercise performance (Messonnier L et al., 2001), and the lactate level is used as an indicator of aerobic exercise capacity to evaluate exercise performance. Blood lactate concentration is an important indicator in various exercise load methods, from aerobic maximal exercise load using a treadmill to short-term high-intensity anaerobic maximal exercise load (Kim Chang-seon et al., Exercise Science 26(4) 239-247, 2017).

[0008] Over the past 50 to 100 years, a decline in human physical activity has been linked to an increased incidence of metabolic diseases such as type 2 diabetes, obesity, and cardiovascular disease. According to the World Health Organization, physical inactivity is the fourth leading cause of death. Due to these trends, organizations such as the World Health Organization, the American Heart Association, and the British Heart Foundation recommend at least 30 minutes of aerobic exercise five or more days per week. Indeed, exercise has been shown to reduce the incidence of diabetes, obesity, breast cancer, and colon cancer, and has been shown to have a beneficial therapeutic effect on depression.

[0009] One of the representative methods for promoting energy consumption to improve fatigue and exercise performance is to increase fatty acid oxidation by mitochondria to generate adenosine triphosphate (ATP).

[0010] Skeletal muscle mitochondria are highly dynamic organelles that undergo significant remodeling in response to various physiological and pathophysiological stresses to meet energy and muscle contraction demands. Changes in muscle energy demands are largely related to changes in physical activity, and sustained endurance exercise training increases the mitochondrial content within muscles.

[0011] Mitochondrial content and oxidative energy metabolism are linked to obesity, diabetes, and aging, but simply increasing mitochondrial biogenesis alone is not sufficient to enhance energy metabolism. This requires a highly regulated system, with quality control mechanisms in muscle, such as autophagy and mitophagy, playing a crucial role in replacing damaged mitochondria.

[0012] Skeletal muscle possesses a highly integrated system that senses and coordinates changes in physical activity or extracellular signals that control mitochondrial biogenesis, and quality control of muscle mitochondria through autophagy and mitophagy mechanisms plays a crucial role in the relationship between disease and aging.

[0013] Mitochondria are closely regulated in function, morphology, and number within the mitochondrial quality control (MQC) system. PGC1α and PGC1β are factors that regulate the MQC system and are involved in mitochondrial biogenesis and mitochondrial energy metabolism by binding to pro-oxidant factors such as NRF1 / NFR2 and TFAM (Lei Chen et al., 2022).

[0014] Meanwhile, human milk oligosaccharides (HMOs) are functionally very important components contained in breast milk, and have been reported to have intestinal lactic acid bacteria activity, prevention of intestinal and respiratory tract harmful microbial infections, immune activation and anti-inflammatory effects, and may also help improve brain and cognitive abilities (Bruggencate S. et al., 2014). Among HMOs, sialylated milk oligosaccharides (SMOs) have the ability to act as decoys that prevent pathogens and pathogen invasion, and are reported to act as receptor mimics of influenza viruses and help protect infants from viruses (Jingyu Yan,. Et al., 2018). There are various types of SMOs, including sialyllactose, Disialyllacto-N-tetraose (DSLNT), 3'-sialyllactosamine (3'SLN), 6'-sialyllactosamine (6'SLN), 3'-sialyl-3-fucosyllactose (3'S3FL), sialyllacto-N-tetraose a (LSTa), sialyllacto-N-tetraose b (LSTb), sialyllacto-N-tetraose c (LSTc), sialyllacto-N-neotetraose d (LSTd), disialyllacto-Nn-tetraose (dsLNnt), disialyllactose (DSL), disialyllacto-Nn-tetraose (dsLNnt), and disialyllactose (DSL), and among these, sialyllactose has the highest content. Sialyllactose is the smallest and most basic oligosaccharide with sialic acid attached, and is classified into 3'-sialyllactose and 6'-sialyllactose based on the structural difference in the sialic acid attached.3'-sialyllactose is found primarily in the milk of most mammals, including cows, and 6'-sialyllactose is found primarily in large quantities in human colostrum (McKeen. S et al., 2019).

[0015] 3'-sialyllactose content did not change significantly throughout the lactation period, but 6'-sialyllactose was present in colostrum three times more than 3'-sialyllactose, and then gradually decreased in content to a level similar to 3'-sialyllactose in mature milk (B Wang and J Brand-Miller, 2003).

[0016] Sialyllactose has been administered to patients with rheumatoid arthritis and has been confirmed to have an effect on treating autoimmune diseases according to changes in IgG. Patents have been confirmed for related diseases such as arthritis or related autoimmune diseases (US 5164374), degenerative arthritis (10-1773066), and a composition that increases the expression of PGC-1α (10-2018-7023369). However, its effect on improving muscle function, such as reducing blood lactate after exercise and improving exercise performance, is not yet known.

[0017]

[0018] The present invention has been devised to solve the problems described above, and the purpose of the invention is to provide a composition for improving muscle function.

[0019] In addition, another object of the present invention is to provide a food composition for improving muscle function.

[0020]

[0021] The technical problems to be solved by the present invention are not limited to the technical problems mentioned above, and other technical problems not mentioned can be clearly understood by a person having ordinary skill in the technical field to which the present invention belongs from the description below.

[0022]

[0023] In order to achieve the above technical task, one embodiment of the present invention provides a composition for improving muscle function comprising sialyloligosaccharide or a derivative thereof.

[0024] In an embodiment of the present invention, the sialyl oligosaccharide may be 3'-sialyllactose, 6'-sialyllactose, sialyllacto-N-tetraose a (LSTa), sialyllacto-N-tetraose b (LSTb), sialyllacto-N-neotetraose c (LSTc), sialyllacto-N-neotetraose d (LSTd), disialyllacto-N-tetraose (DSLT), disialyllacto-N-neotetraose (DSLNnT), or disialyllactose (DSL).

[0025] In an embodiment of the present invention, the muscle function may be anti-fatigue, muscle endurance, or exercise performance ability.

[0026] In an embodiment of the present invention, the improvement in muscle function may be due to a decrease in blood lactate concentration, an increase in glucose concentration, an increase in muscle mass, or an increase in the diameter of muscle fibers.

[0027] In an embodiment of the present invention, the improvement in muscle function may be due to an increase in the expression of mitochondrial complex Ⅰ (NADH:ubiquinone oxidoreductase), mitochondrial complex Ⅲ (ubiquinol-cytochrome c reductase), mitochondrial complex Ⅳ (Cytochrome c oxidase), or mitochondrial complex Ⅴ (ATP synthase).

[0028] In an embodiment of the present invention, the improvement in muscle function may not be due to the expression regulation mechanism of PGC1α and PGC1β.

[0029] Another embodiment of the present invention provides a food composition for improving muscle function comprising sialyloligosaccharide or a food-related acceptable salt thereof.

[0030] In an embodiment of the present invention, the sialyl oligosaccharide may be 3'-sialyllactose, 6'-sialyllactose, sialyllacto-N-tetraose a (LSTa), sialyllacto-N-tetraose b (LSTb), sialyllacto-N-neotetraose c (LSTc), sialyllacto-N-neotetraose d (LSTd), disialyllacto-N-tetraose (DSLT), disialyllacto-N-neotetraose (DSLNnT), or disialyllactose (DSL).

[0031] In an embodiment of the present invention, the muscle function may be anti-fatigue, muscle endurance, or exercise performance ability.

[0032] In an embodiment of the present invention, the improvement in muscle function may be due to a decrease in blood lactate concentration, an increase in blood glucose concentration, an increase in muscle mass, or an increase in the diameter of muscle fibers.

[0033] In an embodiment of the present invention, the improvement in muscle function may be due to an increase in the expression of mitochondrial complex Ⅰ (NADH:ubiquinone oxidoreductase), mitochondrial complex Ⅲ (ubiquinol-cytochrome c reductase), mitochondrial complex Ⅳ (Cytochrome c oxidase), or mitochondrial complex Ⅴ (ATP synthase).

[0034] In an embodiment of the present invention, it may not be based on the expression regulation mechanism of PGC1α and PGC1β.

[0035]

[0036] The above-described problem-solving methods are merely exemplary and should not be construed as limiting the present invention. In addition to the exemplary embodiments described above, additional embodiments may be included in the drawings and detailed description of the invention.

[0037]

[0038] The present invention relates to a composition and a food composition for improving muscle function, which contain sialyllactose as an active ingredient, thereby increasing the expression of genes related to reducing blood lactic acid concentration and improving muscle function, thereby improving muscle function such as anti-fatigue, muscle endurance, and exercise performance without excessively increasing the concentration of lactic acid, which is a fatigue substance.

[0039]

[0040] The effects of the present invention are not limited to the above-described effects, and should be understood to include all effects that can be inferred from the composition of the invention described in the description or claims of the present invention.

[0041]

[0042] Figure 1 shows changes in body weight and food intake following administration of 6'-sialyllactose.

[0043] Figure 2 shows the results of exhaustion treadmill exercise.

[0044] Figure 3 shows the grip strength results.

[0045] Figure 4 shows the blood lactate and glucose concentrations at rest.

[0046] Figure 5 shows the changes in blood lactate concentration and glucose before and after exercise.

[0047] Figure 6 shows the difference in muscle weight between the soleus and gastrocnemius muscles due to 6'-sialyllactose.

[0048] Figure 7 shows the difference in muscle fiber diameter of the gastrocnemius muscle due to 6'-sialyllactose.

[0049] Figure 8 shows the changes in the amount of mitochondrial OXPHOS complex protein in the gastrocnemius muscle.

[0050] Figure 9 shows the change in the amount of PGC1α and PGC1β proteins in the gastrocnemius muscle due to 6'-sarlyllactose.

[0051] Figure 10 shows changes in body weight and food intake according to the composition of sialyllactose.

[0052] Figure 11 shows the difference in exercise performance according to the composition of sialyllactose.

[0053] Figure 12 shows the blood lactate and glucose concentrations according to the composition of sialyllactose.

[0054]

[0055] Hereinafter, the present invention will be described in detail.

[0056]

[0057] The present invention relates to a composition for improving muscle function comprising sialyloligosaccharide or a derivative thereof.

[0058] The above sialyl oligosaccharides are in the form of one or two sialic acids attached to oligosaccharide residues such as lactose, lactene tetraose, or lactene neotetraose, for example, 3'-sialyllactose, 6'-sialyllactose, sialyllacto-N-tetraose a (LSTa), sialyllacto-N-tetraose b (LSTb), sialyllacto-N-neotetraose c (LSTc), sialyllacto-N-neotetraose d (LSTd), disialyllacto-N-tetraose (DSLT), It may be any one selected from disialyllacto-N-neotetraose (DSLNnT) and disialyllactose (DSL), or a mixture thereof.

[0059] In the present invention, the term 'derivative' means a compound in which the functional group of a sialyl oligosaccharide is changed by introduction, substitution, oxidation, reduction, etc., without significantly changing the structure and properties of the parent compound. There is no limitation on the type of such functional group, and for example, it may include, but is not limited to, a C1 to C20 bicyclic hydrocarbon group independently substituted or unsubstituted with a hydroxyl group, a phenoxy group, a thienyl group, a furyl group, a pyridyl group, a cyclohexyl group, an alkyl alcohol group, an alkyl dialcohol group, or a substituted or unsubstituted phenyl; a C3 to C30 cyclic hydrocarbon group substituted or unsubstituted with a hydroxyl group, a hydroxymethyl group, a methyl group, or an amino group; or a sugar moiety.

[0060] The aforementioned muscle function may be, for example, anti-fatigue, muscle endurance, or exercise performance. Specifically, the present invention can prevent muscle fatigue, improve muscle endurance, prevent muscle endurance decline, and enhance muscle exercise performance.

[0061] The above improvement in muscle function may be due to a decrease in blood lactate concentration, an increase in blood glucose concentration, an increase in muscle mass, or an increase in muscle fiber diameter.

[0062] Blood lactate concentrations remain relatively low at rest, but high-intensity physical activity converts pyruvate to lactic acid through glycolysis within muscle cells. This lactic acid releases hydrogen ions within the muscle cells and is converted to lactate, which changes the body's pH from slightly alkaline to acidic. Most of the lactic acid enters the liver through the blood and is converted to glucose through the Cori cycle, or some is oxidized by the heart and slow-twitch muscle fibers. However, when blood lactate concentrations increase due to high-intensity exercise, the body's pH decreases, which impairs the activity of intracellular enzymes and reduces the energy synthesis required for physical activity, thereby reducing exercise performance. However, sialyl oligosaccharides can reduce blood lactate concentrations, thereby reducing muscle fatigue and improving exercise performance.

[0063] Furthermore, glucose is an energy source produced from liver glycogen. Not only the accumulation of metabolites but also a decrease in glucose levels can cause fatigue. Endurance exercise demands high levels of energy and consumes glucose from liver and muscle tissue glycogen. By increasing blood glucose levels, sialyl oligosaccharides can reduce muscle fatigue and improve exercise performance.

[0064] The above improvement in muscle function may be due to increased expression of mitochondrial complex Ⅰ (NADH:ubiquinone oxidoreductase), mitochondrial complex Ⅲ (ubiquinol-cytochrome c reductase), mitochondrial complex Ⅳ (Cytochrome c oxidase), or mitochondrial complex Ⅴ (ATP synthase).

[0065] Specifically, in mitochondria, ATP is formed through oxidative phosphorylation, activating intracellular energy metabolism, and energy-producing complexes (complexes I, II, III, IV, V) are involved in this process. Specifically, complex I oxidizes NADH, and complex II oxidizes FADH2 to reduce ubiquinol, which is then oxidized in complex III to reduce cytochrome c, and complex IV oxidizes cytochrome c and transfers electrons to molecular oxygen to produce water. In this process, hydrogen ions from the inner mitochondrial membrane flow out of the mitochondrial matrix, creating a membrane potential difference, and when hydrogen ions flow back into the mitochondrial matrix by complex V, ATP is produced from ADP. More specifically, the present invention can improve muscle function by increasing the expression of complex Ⅰ-NDUF88, complex Ⅲ-UQCRC2 (Ubiquinol-Cytochrome C Reductase Core Protein 2), complex Ⅳ-MTCO1 (Mitochondrially Encoded Cytochrome C Oxidase I), and complex Ⅴ-ATP5.

[0066] Meanwhile, in one embodiment of the present invention, it was found that sialyllactose does not affect the increase in the expression of PGC1α (Peroxisome proliferator-activated receptor gamma coactivator 1-alpha) and PGC1β (Peroxisome proliferator-activated receptor gamma coactivator 1-beta). PGC1α and PGC1β are proteins that regulate mitochondrial synthesis and function, and are known to regulate muscle function by participating in energy production through oxidative phosphorylation and mitochondrial biogenesis. However, in the present invention, the improvement in muscle anti-fatigue, exercise performance, muscle mass increase, and OXPHOS increase by sialylligosaccharide may not be due to the expression regulation mechanism of PGC1α and PGC1β.

[0067] The composition of the present invention may further comprise human milk oligosaccharide, if necessary.

[0068] The human breast milk derived oligosaccharide may be, for example, any one selected from among lacto-N-tetraose, lacto-N-neotetraose, 2'-fucosyllactose, 3-fucosyllactose, sialyllacto-N-tetraose a, sialyllacto-N-tetraose b, sialyllacto-N-neotetraose c, sialyllacto-N-neotetraose d and lacto-N-fucopentaose, or a combination thereof.

[0069] The composition of the present invention may contain various additives, such as isotonic agents, excipients, diluents, thickeners, stabilizers, buffers, and preservatives, depending on the form of use. The blending amounts of these additives may be appropriately set depending on the form of use of the composition of the present invention.

[0070] The composition of the present invention can be implemented in various composition forms, such as, for example, a pharmaceutical composition, a quasi-drug composition, a feed (feed additive) composition, etc.

[0071] The pharmaceutical composition may further comprise a pharmaceutically acceptable carrier.

[0072] The pharmaceutically acceptable carrier means any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. Examples of pharmaceutically acceptable carriers include water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, and the like, and mixtures thereof. In many cases, it will be desirable to include isotonic agents, such as sugars, polyhydric alcohols, such as mannitol, sorbitol, or sodium chloride, in the composition. Additional examples of pharmaceutically acceptable substances include wetting agents or minimal amounts of auxiliary substances, such as wetting or emulsifying agents, preservatives, or buffers, which extend or enhance the shelf life or efficacy of the antibody.

[0073] The compositions of the present invention may take various forms, for example, liquid, semi-solid and solid dosage forms, for example, liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes and suppositories. The preferred form will depend on the intended mode of use. The compositions of the present invention may take the form of suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulating agents, for example, suspending, stabilizing and / or dispersing agents. Alternatively, the active ingredient may be in powder form for use with a suitable vehicle, for example, a sterile, pyrogen-free substance, prior to use.

[0074] In addition, the composition of the present invention can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, or sterile injection solutions, respectively, according to conventional methods. Specifically, when formulating, it can be prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants that are commonly used. Solid preparations for oral administration include, but are not limited to, tablets, pills, powders, granules, and capsules. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. In addition to liquids for oral administration and liquid paraffin, it can be prepared by adding various excipients such as wetting agents, sweeteners, fragrances, and preservatives. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solutions and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include withepsol, macrosol, Tween 61, cocoa butter, laurin, and glycerogelatin.

[0075] The term “quasi-drug” used in the present invention means an article other than a device, machine or apparatus used for the purpose of diagnosing, treating, alleviating, managing or preventing a disease of a human or animal, and an article other than a device, machine or apparatus used for the purpose of exerting a pharmacological effect on the structure and function of a human or animal, and may include, but is not limited to, an oral preparation, and the method of formulation, dosage, method of use, components, etc. of a quasi-drug may be appropriately selected from conventional techniques known in the art.

[0076] In addition to the above ingredients, the above-mentioned pharmaceutical composition may further include a pharmaceutically acceptable carrier, excipient, or diluent, as needed. The pharmaceutically acceptable carrier, excipient, or diluent is not limited as long as it does not impair the effects of the present invention, and may include, for example, fillers, bulking agents, binders, wetting agents, disintegrants, surfactants, lubricants, sweeteners, fragrances, preservatives, etc.

[0077] The above feed composition can be formulated in the form of a conventional feed and can include known feed ingredients. In addition, it can also be used as a feed additive added in the form of an additive to the feed being used. The feed additive of the present invention corresponds to a supplementary feed under the Feed Management Act, and may additionally include mineral preparations such as sodium bicarbonate (sodium bicarbonate), bentonite, magnesium oxide, and complex minerals; mineral preparations that are trace minerals such as zinc, copper, cobalt, and selenium; vitamins such as carotene, vitamin E, vitamins A, D, E, nicotinic acid, and vitamin B complex; protected amino acids such as methionine and lysate; protected fatty acids such as fatty acid calcium salts; live bacteria such as probiotics (lactic acid bacteria), yeast cultures, and mold fermentations; and yeast agents.

[0078] The above feed or feed additive can be applied to a number of animal diets including mammals, poultry and fish.

[0079]

[0080] The present invention relates to a food composition for improving muscle function comprising sialyloligosaccharide or a food-wise acceptable salt thereof.

[0081] The mechanism of the above-mentioned sialyl oligosaccharide and improvement of muscle function is as described above.

[0082] The food composition of the present invention may include conventional food additives, and may include food raw materials usable as food described in the standards and specifications for food commonly used in food manufacturing ('Food Code'), and food additives described in the Food Additive Code.

[0083] The items listed in the above food additive code include, but are not limited to, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, high-molecular-weight pigment, and guar gum; and mixed preparations such as sodium L-glutamate preparations, alkaline agents added to noodles, preservative preparations, and tar color preparations.

[0084] The food composition of the present invention can be manufactured and processed into food in the form of tablets, capsules, powders, granules, liquids, pills, etc. for the purpose of improving muscle function.

[0085] A food composition in tablet form can be prepared by mixing sialyl oligosaccharides with excipients, binders, disintegrants, and other additives, granulating the mixture using a conventional method, adding a lubricant, etc., and then compression molding, or by directly compression molding the mixture. In addition, the health functional food in tablet form can contain a flavoring agent, etc., as needed.

[0086] Among the food compositions in capsule form, hard capsules can be manufactured by filling a mixture of the sialyl oligosaccharide of the present invention with additives such as excipients into a conventional hard capsule, and soft capsules can be manufactured by filling a mixture of the sialyl oligosaccharide of the present invention with additives such as excipients into a capsule base such as gelatin. The soft capsules may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative, etc., as necessary.

[0087] The food composition in the form of a ring can be prepared by molding a mixture of the sialyl oligosaccharide of the present invention and an excipient, a binder, a disintegrant, etc., using a conventionally known method, and, if necessary, can be coated with white sugar or another coating agent, or the surface can be coated with a material such as starch or talc.

[0088] A granular food composition can be manufactured into a granular form by mixing a mixture of the sialyl oligosaccharide excipient, binder, disintegrant, etc. of the present invention using a conventionally known method, and may contain a flavoring agent, a flavoring agent, etc., as needed.

[0089] The above food composition may include beverages, meat, chocolate, food, confectionery, pizza, ramen, other noodles, gum, candy, ice cream, alcoholic beverages, vitamin complexes, and health supplements.

[0090]

[0091] Hereinafter, the present invention will be described in detail by way of examples to specifically explain the present invention.

[0092]

[0093] 1. Confirmation of the efficacy of 6'-Sialyllactose on exercise capacity

[0094] 1.1 Experimental method

[0095] Four-week-old male C57BL / 6J mice (Central Laboratory Animal Co., Ltd.) were acclimatized for one week under conditions of temperature 23±2℃ and humidity 50±10%, and food and water were provided ad libitum. The study was approved by the Animal Experiment Ethics Committee of Chungnam National University.

[0096] In order to measure changes in exercise performance according to the administration of 6'-sialyllactose, first, mice acclimatized in the above process were randomly selected, and 8 mice were selected for the experimental group and 8 mice for the control group. Then, 6'-sialyllactose 100 mg / kg / day or saline (control group) was orally administered 5 times a week for 12 weeks. Body weight and food intake were measured once a week.

[0097] Grip strength was measured at weeks 3 and 10, an exhaustion test was performed at week 10, and a simple exhaustion test was performed at week 12. The recovery group was observed. Afterwards, autopsies were performed to determine muscle weight and biochemical analyses.

[0098]

[0099] 1.2 Changes in weight and diet

[0100] After 12 weeks of administration of 6'-sialyllactose or saline, there was no difference in body weight between the experimental and control groups (Fig. 1A). This was also confirmed by weight gain (Fig. 1B). To confirm the difference in diet, the same amount of food was provided, and the remaining food amount was measured one week later to determine the actual food intake and the change in the amount (Fig. 1C). As a result of checking body weight and food intake over 12 weeks, it was confirmed that there was no change in body weight and food intake between the experimental and control groups.

[0101]

[0102] 1.3 Assessment of exercise performance through exhaustion treadmill exercise and grip strength

[0103] To assess exercise capacity, exhaustion treadmill exercise was performed after 10 weeks of administration. The treadmill used was manufactured in-house, and exercise was performed at a 16° incline with an electrical gap of less than 4 times per minute to ensure that the electrical stimulation was not too strong. After acclimatizing for 3 minutes at a speed of 16.67 m / min, the speed was increased by 1.67 m / min every 3 minutes until exhaustion was reached, and the exercise capacity of each group of mice was assessed.

[0104] As a result of measuring the total treadmill distance, it was confirmed that the total treadmill distance increased statistically significantly (P<0.05) in the 6'-sialyllactose administration group compared to the control group (Fig. 2A).

[0105] The time until exhaustion was measured at this time also achieved statistical significance (P<0.05) (Fig. 2B). Converting this to total work volume, calculated by multiplying body weight and total distance traveled, also achieved statistical significance (P<0.01), confirming that the 6'-sialyllactose group was able to travel a greater distance (Fig. 2C).

[0106] When grip strength was measured and the rate of increase was confirmed at weeks 3 and 10, it was confirmed that the increase was 12.3% greater in the 6'-sialyllactose group than in the control group (Fig. 3).

[0107] When these results are combined, it can be seen that the group that took 6'-sialyllactose was able to run longer distances and better than the control group.

[0108]

[0109] 1.4 Changes in blood lactate and glucose concentrations before and after the exhaustion experiment

[0110] After 12 weeks of administration, a small amount of blood was collected from the tail vein to measure the blood lactate and glucose concentrations of the experimental and control groups in a stable state. Lactate concentration was measured using an Arkray lactatepro portable lactate meter, and glucose concentration was measured using an Accu-Chek blood glucose meter.

[0111] In order to check the blood lactate and glucose concentrations before and after exercise, the mice were allowed to acclimate at a speed of 16.67 m / min for 3 minutes, and then the speed was increased by 1.67 m / min every 3 minutes for a total of 20 minutes. After running on the treadmill, a small amount of blood was collected from the tail vein of the mice before exercise, immediately after exercise, and 5 and 10 minutes after exercise, and the blood lactate and glucose concentrations were measured using a measuring strip.

[0112] As a result, it was confirmed that the blood lactate and glucose concentrations in the experimental group were statistically significantly lower than those in the control group without exercise (Fig. 4).

[0113] The blood lactate concentration of the experimental group was statistically significantly lower than that of the control group not only before exercise (Rest) but also immediately after exercise and 10 minutes after exercise (Figure 5A).

[0114] Through this, it was confirmed that the experimental group that took 6'-sialyllactose maintained a lower blood lactate concentration than the control group that did not take it, and that the blood lactate concentration, a fatigue substance, was significantly lower even immediately after intense exercise.

[0115] Additionally, in the control group that did not take 6'-sialyllactose, there was no statistically significant change in the glucose concentration before and after exercise, but in the experimental group, the blood glucose concentration, which was low when not exercising, rose sharply immediately after exercise. This increased with statistical significance compared to the control group at 5 and 10 minutes (Fig. 5B).

[0116] The blood glucose concentration of the actual experimental group was statistically significantly increased 5 and 10 minutes after exercise compared to before exercise (Figure 5B).

[0117] In summary, it was confirmed that the experimental group that took 6'-sialyllactose had the effect of quickly increasing the required glucose and keeping the lactic acid concentration that induces fatigue low, thereby preventing the accumulation of fatigue substances.

[0118]

[0119] 1.5 Changes in muscle weight

[0120] The soleus muscle is a powerful muscle located at the back of the leg (calf). It is closely related to the gastrocnemius muscle. It is essential for walking, running, and maintaining balance. It is also a major factor influencing walking speed in older adults who meet the diagnostic criteria for sarcopenia. The soleus muscle has a higher proportion of slow-twitch (slow-twitch) fibers than many other muscles, making it highly fatigue-resistant. However, it can become easily fatigued if damaged.

[0121] The gastrocnemius is a muscle frequently used in sprinting and jumping. It participates in knee flexion and extension, and acts as an agonist or antagonist of the hamstring muscles. It is a key muscle in osteoarthritis. Furthermore, the gastrocnemius is prone to thigh spasms, and is prone to contractions when fatigued.

[0122] The weights of the soleus and gastrocnemius muscles were measured during sacrifice of the experimental and control groups that received oral 6'-sialyllactose for 12 weeks.

[0123] As a result, it was confirmed that the gastrocnemius muscle increased by 6.1% in the experimental group compared to the control group, and the soleus muscle increased by 17.9% in the experimental group compared to the control group, which was a statistically significant increase (P<0.05) (Fig. 6).

[0124]

[0125] 1.6. Changes in muscle fiber diameter

[0126] Fiber diameter size of the gastrocnemius muscle was measured using H&E staining, and the distribution ratio was calculated (Fig. 7A). In the 6'-sialyllactose group, the fiber diameter size ratio tended to shift toward larger sizes compared to the control group (Fig. 7B). In addition, the mean Min. Ferret Diameter was statistically significantly (P<0.0001) increased in the 6'-sialyllactose group (Fig. 7C).

[0127]

[0128] 1.7 Biochemical changes in muscles

[0129] (1) Changes in mitochondrial oxidative phosphorylation

[0130] The gastrocnemius muscles of the experimental group that took 6'-sialyllactose for 12 weeks and the control group that took only saline were isolated, and cell lysates were prepared using T-PER Tissue Protein Extraction Reagent buffer. Proteins were isolated and quantified using Quick Start™ Bradford Protein Assay (Bio-rad). Equal amounts of protein were loaded onto 4-20% Mini-PROTEAN®TGX™ precast protein gels (Bio-rad), subjected to SDS-PAGE, transferred to PVDF membranes, blocked, and then subjected to anti-OXPHOS complexes reaction. After overnight reaction, secondary antibody reaction was performed and detection was performed with ECL reagent (Bio-rad).

[0131] As a result, it was confirmed that almost all of the mitochondrial energy metabolism of the group that took 6'-sialyllactose was activated statistically significantly more than the control group (Fig. 8). It was found that 6'-sialyllactose significantly increased Complex V (CV-ATP5A), the final complex of oxidative phosphorylation that produces ATP from ADP, Complex IV, the complex just before producing water using oxygen, Complex I, the first step of oxidative phosphorylation, and Complex III, the second step.

[0132] (2) PGC1α and PGC1β

[0133] It was confirmed that there was no statistically significant difference in the amount of PGC1α and PGC1β proteins in the group administered 6'-sialyllactose compared to the control group (Fig. 9).

[0134] Through this, it was confirmed that the increase in muscle mass, decrease in lactic acid, and increase in exercise performance induced by 6'-sialyllactose did not go through PGC1α or PGC1β.

[0135]

[0136] 2. Confirmation of exercise performance efficacy according to sialyllactose composition

[0137] 2.1 Experimental method

[0138] Four-week-old male C57BL / 6J mice (Central Laboratory Animal Co., Ltd.) were acclimatized for one week under conditions of temperature 23±2℃ and humidity 50±10%, and food and water were provided ad libitum. The study was approved by the Animal Experiment Ethics Committee of Pohang University of Science and Technology.

[0139] In order to measure changes in exercise performance according to the administration of sialyllactose, first, the mice acclimatized in the above process were randomly selected, and 12 mice were selected for the experimental group and 12 for the control group. Then, 100 mg / kg / day of 3'-sialyllactose, 100 mg / kg / day of 6'-sialyllactose, 100 mg / kg / day of a mixture of 3'-sialyllactose and 6'-sialyllactose (1:3), or saline (control group) were orally administered three times a week for 9 weeks. Body weight and food intake were measured once a week.

[0140] Grip strength was measured and an exhaustion test was conducted in the 8th week, and a simple exhaustion test was conducted in the 9th week, and observations were conducted on the recovery group.

[0141]

[0142] 2.2 Changes in weight and diet

[0143] After 9 weeks of administration of sialyllactose or saline, there was no difference in body weight between the experimental and control groups (Fig. 10A). This was also confirmed by weight gain (Fig. 10B). To confirm the difference in diet, the same amount of food was provided, and the remaining food amount was measured one week later to determine the actual food intake and confirm the change (Fig. 10C). As a result of checking the food intake for 9 weeks, a statistically significant difference was confirmed between the 6'-sialyllactose group and the control group.

[0144]

[0145] 2.3 Evaluation of exercise performance through exhaustion treadmill exercise and grip strength

[0146] To determine exercise capacity, exhaustion treadmill and grip strength were measured after 8 weeks of administration.

[0147] The treadmill used in the exhaustion treadmill was self-made, and exercise was performed at a level where the electrical stimulation was not strong by setting the electrical gap to less than 4 times per minute at a 16° incline. After acclimating for 3 minutes at a speed of 16.67 m / min, the speed was increased by 1.67 m / min every 3 minutes until exhaustion was achieved, and the exercise ability of each group of mice was determined.

[0148] As a result of measuring the total treadmill distance, it was confirmed that the total treadmill distance increased statistically significantly in the 3'-sialyllactose only group, the 6'-sialyllactose only group, and the 3'-sialyllactose and 6'-sialyllactose mixture group compared to the control group (Fig. 11A).

[0149] The results of measuring the time until exhaustion at this time also secured statistical significance (Fig. 11B).

[0150] A maximal voluntary force testing system was used to measure the grip strength of the mouse forelimbs at week 8. As a result, statistically significant increases were observed in the 3'-sialyllactose alone, 6'-sialyllactose alone, and 3'-sialyllactose and 6'-sialyllactose mixture groups at week 8 (Fig. 11C).

[0151]

[0152] 2.4 Changes in blood lactate and glucose concentrations before and after the exhaustion experiment

[0153] In order to confirm muscle fatigue before and after the exhaustion experiment, changes in blood lactate concentration were checked. In order to measure the blood lactate concentration of the experimental and control groups before and after exercise after 9 weeks of administration, the speed was adjusted to 16.67 m / min for 3 minutes, and then the speed was increased by 1.67 m / min every 3 minutes for a total performance time of 20 minutes. After that, a small amount of blood was collected from the tail vein of the mice before exercise, immediately after exercise, and 5, 10, and 15 minutes after exercise. Lactate concentration was measured using an Arkray lactatepro portable lactate meter, and glucose concentration was measured using an Accu-Chek blood glucose meter.

[0154] As a result, in the control group, the blood lactate concentration increased rapidly immediately after exercise (0 min) and then maintained a similar concentration until the 15-minute recovery period. In the group that took 3'-sialyllactose alone or 6'-sialyllactose alone, the lactate concentration increased immediately after exercise and gradually decreased at 5 and 10 minutes of the recovery period, and at 15 minutes of the recovery period, it decreased to a level similar to before exercise (Rest). On the other hand, in the group that took a mixture of 3'-sialyllactose and 6'-sialyllactose, the lactate concentration did not increase significantly immediately after exercise and showed a level similar to before exercise (Rest), which was confirmed to continue until the 15-minute recovery period (Fig. 12A).

[0155] In the case of blood glucose, the blood glucose concentration in the group taking the mixture of 3'-sialyllactose and 6'-sialyllactose was found to be maintained at a significantly higher level from immediately after exercise (0 min) to the 15 min recovery period compared to the control group (Figure 12B).

[0156] When 3'-sialyllactose alone, 6'-sialyllactose alone, and a mixture of 3'-sialyllactose and 6'-sialyllactose were administered to mice, it was confirmed that there was no difference in exercise performance between single administration and mixed administration. Meanwhile, in the case of blood lactate concentration and glucose concentration, it was confirmed that mixed administration of sialyllactose was more effective in lowering the blood lactate concentration, a fatigue substance immediately after exercise, and also increasing blood glucose, an energy source, and improving exercise performance than administration of sialyllactose alone.

Claims

1. A composition for improving muscle function comprising sialyloligosaccharide or a derivative thereof.

2. In claim 1, the sialyl oligosaccharide is 3'-sialyllactose, 6'-sialyllactose, sialyllacto-N-tetraose a (LSTa), sialyllacto-N-tetraose b (LSTb), sialyllacto-N-neotetraose c (LSTc), sialyllacto-N-neotetraose d (LSTd), disialyllacto-N-tetraose (DSLT), disialyllacto-N-neotetraose (DSLNnT), or disialyllactose (DSLNnT). A composition for improving muscle function (DSL).

3. A composition for improving muscle function according to claim 1, wherein the muscle function is anti-fatigue, muscle endurance or exercise performance ability.

4. A composition for improving muscle function according to claim 1, wherein the improvement in muscle function is due to a decrease in blood lactic acid concentration, an increase in blood glucose concentration, an increase in muscle mass, or an increase in the diameter of muscle fibers.

5. A composition for improving muscle function according to claim 1, wherein the improvement in muscle function is due to an increase in the expression of mitochondrial complex Ⅰ (NADH:ubiquinone oxidoreductase), mitochondrial complex Ⅲ (ubiquinol-cytochrome c reductase), mitochondrial complex Ⅳ (Cytochrome c oxidase), or mitochondrial complex Ⅴ (ATP synthase).

6. A composition for improving muscle function according to claim 1, wherein the improvement in muscle function is not due to a mechanism for regulating the expression of PGC1α and PGC1β.

7. A food composition for improving muscle function comprising sialyloligosaccharide or a food-related acceptable salt thereof.

8. In claim 7, the sialyl oligosaccharide is 3'-sialyllactose, 6'-sialyllactose, sialyllacto-N-tetraose a (LSTa), sialyllacto-N-tetraose b (LSTb), sialyllacto-N-neotetraose c (LSTc), sialyllacto-N-neotetraose d (LSTd), disialyllacto-N-tetraose (DSLT), disialyllacto-N-neotetraose (DSLNnT), or disialyllactose (DSLNnT). A food composition for improving muscle function (DSL).

9. A food composition for improving muscle function according to claim 7, wherein the muscle function is anti-fatigue, muscle endurance or exercise performance ability.

10. A food composition for improving muscle function according to claim 7, wherein the improvement in muscle function is due to a decrease in blood lactic acid concentration, an increase in blood glucose concentration, an increase in muscle mass, or an increase in the diameter of muscle fibers.

11. A food composition for improving muscle function according to claim 7, wherein the improvement in muscle function is due to an increase in the expression of mitochondrial complex Ⅰ (NADH:ubiquinone oxidoreductase), mitochondrial complex Ⅲ (ubiquinol-cytochrome c reductase), mitochondrial complex Ⅳ (Cytochrome c oxidase), or mitochondrial complex Ⅴ (ATP synthase).

12. A food composition for improving muscle function according to claim 7, wherein the improvement in muscle function is not due to a mechanism for regulating the expression of PGC1α and PGC1β.

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