Composition containing extract of Phellinus linteus GKPl for improving athletic performance and reducing exercise fatigue

Abstract

To provide a composition for improving exercise performance and reducing exercise fatigue, containing an extract of Phellinus linteus GKPl.SOLUTION: The present invention relates to a composition for improving exercise performance and reducing exercise fatigue, which contains an extract of Phellinus linteus GKPl. The composition contains the extract of Phellinus linteus GKPl as an active ingredient, the extract containing a hot water extract and / or an ethanol extract, thereby improving exercise duration and a glycogen content in the liver and muscles, reducing contents of lactic acid and urea nitrogen in blood after exercise, so as to improve exercise performance and reduce exercise fatigue.SELECTED DRAWING: Figure 5

Description

[Technical Field]

[0001] The present invention relates to a composition for improving exercise performance and reducing exercise fatigue, and more particularly to a composition for improving exercise performance and reducing exercise fatigue containing an extract of Phellinus linteus GKPl. [Background technology]

[0002] Increasingly, research is being conducted focusing on exercise science (e.g., exercise psychology, exercise physiology, exercise biomechanics, exercise nutrition, and traditional supportive therapies) to improve athletes' athletic performance in sports competitions. Factors that affect athletic performance include physical ability, athletic skill, vision, fatigue level, illness, athletic injury, exercise nutrition, psychological factors, and external factors. Among these, exercise nutrition not only plays an important role in building body composition and providing sufficient energy, but is also one of the most important keys to improving metabolic rate, body recovery speed, and athletes' immunity, and / or reducing the risk of athletic injury. Summary of the Invention [Problem to be solved by the invention]

[0003] In addition to the ratio and timing of intake of carbohydrates, lipids, proteins, vitamins, minerals, and fluids, athletic supplements (e.g., creatine, caffeine) can also improve athletic performance and / or reduce exercise fatigue. However, some athletic supplements contain active substances that may cause side effects and ultimately harm the overall health of athletes, and are therefore classified as prohibited substances. Therefore, the search for safe and effective active substances in athletic supplements has become an important direction of research.

[0004] Phellinus linteus is a medicinal fungus belonging to the Phellinaceae family. Phellinus linteus is highly safe and has antioxidant, anti-cancer, anti-dementia, anti-inflammatory, anti-allergic (e.g., allergic rhinitis, eczema, rheumatoid arthritis), liver protection, cardiovascular disease prevention, improved sleep quality, pain relief, uric acid suppression, and skin care properties. However, there has been little research into the effects of Phellinus linteus on improving exercise performance and reducing exercise fatigue. [Means for solving the problem]

[0005] Therefore, one aspect of the present invention provides a composition for improving exercise performance, which contains an extract of Meshimakobu (Phellinus linteus) GKP1, so as to reduce the urea nitrogen content in the blood of a subject after exercise.

[0006] Another aspect of the present invention provides an oral composition for reducing exercise fatigue, which contains an extract of Phellinus linteus GKP1, so as to reduce the urea nitrogen content in the blood after exercise.

[0007] According to the above aspect of the present invention, there is provided a composition for improving athletic performance containing an extract of Phellinus linteus GKPl, the composition comprising an extract of Phellinus linteus GKPl as an active ingredient, and the extract comprising a hot water extract and / or an ethanol extract, thereby improving glycogen content in the liver and / or muscle. The extract of Phellinus linteus GKPl was deposited at the Bioresource Collection and Research Center (BCRC) of the Food Industry Research and Development Institute (FIRDI), Taiwan on July 18, 2019, under the accession number BCRC 930210.

[0008] In one embodiment of the present invention, the hot water extract is obtained by subjecting mycelia of Phellinus linteus GKP1 to hot water extraction treatment with water at 80°C to 120°C for 20 to 40 minutes.

[0009] In one embodiment of the present invention, the ethanol extract is obtained by subjecting the mycelium of Phellinus linteus GKP1 to ethanol extraction.

[0010] In one embodiment of the invention, the composition is a pharmaceutical composition or a food composition.

[0011] In one embodiment of the present invention, the composition may optionally include pharmaceutically and food acceptable carriers, excipients and / or additives.

[0012] In one embodiment of the present invention, subjects administered with an extract of Phellinus linteus GKPl have increased exercise duration compared to control subjects not administered with an extract of Phellinus linteus GKPl, wherein both the control subjects and the subjects do not have muscle atrophy.

[0013] According to another aspect of the present invention, there is provided an oral composition for reducing exercise fatigue, comprising an extract of Phellinus linteus GKPl, which contains an effective dose of an extract of Phellinus linteus GKPl as an active ingredient, the extract comprising a hot water extract and / or an ethanol extract of Phellinus linteus GKPl, and the accession number of Phellinus linteus GKPl is BCRC 930210, thereby reducing the blood urea nitrogen content of a subject after exercise after continuous administration to the subject for at least 28 days.

[0014] In one embodiment of the present invention, when the subject is an adult, the effective dosage is 0.1 g / 60 kg body weight (bw) / day to 2.0 g / 60 kg bw / day.

[0015] In one embodiment of the present invention, when the subject is a mouse, the effective dosage is 0.1 g / kg bw / day to 0.5 g / kg bw / day.

[0016] In one embodiment of the present invention, the subjects administered with the extract of Phellinus linteus GKPl have a reduced blood lactic acid content after exercise compared to the control subjects not administered with the extract of Phellinus linteus GKPl, and both the control subjects and the subjects do not have muscle atrophy. [Effects of the Invention]

[0017] The extract of Phellinus linteus GKPl of the present invention can be used to improve the glycogen content in the liver and / or muscles and reduce the urea nitrogen content in the blood of a subject after exercise, thereby improving exercise performance and / or reducing exercise fatigue, and therefore can be used as an active ingredient to prepare various compositions. [Brief explanation of the drawings]

[0018] To make the above and other objects, features, advantages and embodiments of the present invention more comprehensible, reference is made to the accompanying drawings in which: [Figure 1] 1 is a bar graph showing the weighted swimming exhaustion time of mice in the control group and the experimental group shown in Table 1. [Figure 2] 1 is a bar graph showing the treadmill exhaustion times of mice in the control and experimental groups shown in Table 2. [Figure 3A] 1 is a bar graph showing the increase rate of lactate in the blood of mice in the control group and the experimental group shown in Table 3. [Figure 3B] 1 is a bar graph showing the lactate removal rate in the blood of mice in the control and experimental groups shown in Table 3. [Figure 4] 1 is a bar graph showing the urea nitrogen content in the blood of mice in the control and experimental groups shown in Table 4 before and after long-term swimming. [Figure 5] 1 is a bar graph of glycogen content in liver and muscle of mice in the control and experimental groups shown in Table 5. DETAILED DESCRIPTION OF THE INVENTION

[0019] If the definition or usage of a term in a cited reference is inconsistent with or contrary to the definition of that term herein, the definition in the cited reference shall prevail and the definition in this specification shall prevail. Also, unless otherwise defined by context, singular terms may include pluralities and plural terms may include the singular.

[0020] As described above, the present invention provides a composition for improving exercise performance and reducing exercise fatigue, which contains an extract of Phellinus linteus GKPI. The extract of Phellinus linteus GKPI can improve the glycogen content in the muscle and liver of a subject and reduce the lactic acid and urea nitrogen content in the subject's blood after exercise, thereby improving exercise performance and / or reducing exercise fatigue.

[0021] As used herein, a "subject" refers to a healthy individual. In some examples, the subject is a healthy athlete and / or other healthy individual with a training regimen. In some embodiments, the subject does not have a muscle disorder. In some embodiments, muscle disorders may include, but are not limited to, muscle atrophy and / or sarcopenia.

[0022] As used herein, "athletic performance" refers to the speed, strength, skill, and / or coordination exhibited by a subject during exercise. Different types of exercise may assess athletic performance in different ways. In some embodiments, athletic performance includes muscular endurance and / or cardiorespiratory endurance. In some embodiments, athletic performance may be assessed by exercise duration and / or glycogen content in the liver and muscles. In some embodiments, exercise duration may be assessed by weighted swimming exhaustion time and / or treadmill exhaustion time.

[0023] "Exercise fatigue" as used herein refers to the subject's level of fatigue during exercise, after exercise, and after resting after exercise. In some embodiments, exercise fatigue may be assessed by measuring the lactic acid and / or urea nitrogen content in the blood after exercise.

[0024] Glycogen, a polysaccharide formed by the dehydration and condensation of glucose, is one of the substances used by animals to store energy. It can be produced and stored in liver cells and muscle cells. When muscles cannot readily obtain sufficient energy from blood glucose, they directly decompose glycogen in the cells to provide the energy needed for muscle exercise. Furthermore, when blood glucose levels drop, liver cells also directly decompose glycogen in the cells to replenish blood glucose levels. Therefore, glycogen content in the liver and muscles can affect exercise endurance and improve athletic performance.

[0025] When the energy produced by aerobic respiration is insufficient to meet the energy needs of muscles, the muscles obtain sufficient energy through glycolysis, producing the by-products lactic acid and hydrogen ions. Lactic acid can be transported to the liver via the blood circulation, where it is removed by the liver through the Cori cycle and glucose can be produced. Hydrogen ions can also be removed through the above process. However, if the rate of lactic acid production exceeds the rate of removal, large amounts of lactic acid and hydrogen ions accumulate in the muscles, thereby lowering the muscle pH and affecting the energy transfer of enzymes, resulting in symptoms of exercise fatigue such as muscle pain, heaviness, and fever. Therefore, the amount of lactic acid in the blood can be used as an indicator of exercise fatigue.

[0026] The order in which the body uses energy is carbohydrates (e.g., glucose and / or glycogen), fat, and protein. Therefore, after prolonged exercise, if sufficient energy cannot be supplied through the breakdown of sugars or fats, the muscles will break down proteins and amino acids to generate energy, and the liver will produce urea nitrogen. Urea nitrogen is transported by the blood to the kidneys and then excreted from the body via urine. Therefore, the blood urea nitrogen (BUN) content can also be used as an indicator of exercise fatigue.

[0027] In some embodiments, the accumulation of lactate and / or urea nitrogen after exercise may be assessed by comparing the difference in the lactate and / or urea nitrogen content in the blood before and after exercise, respectively. In other embodiments, the removal of lactate from the blood after exercise and rest may be assessed by comparing the difference in the lactate content in the blood after a short period of exercise and rest, respectively.

[0028] It should be noted that the "exercise" described herein may be divided into aerobic exercise and anaerobic exercise. Aerobic exercise is moderate-intensity exercise that lasts for a long period of time and requires steady, regular muscle contraction and relaxation. Energy in aerobic exercise is primarily generated by the breakdown of sugars or fats through aerobic respiration. In some embodiments, aerobic exercise may include, but is not limited to, long-distance running, walking, cycling, swimming, aerobic boxing, and / or aerobic dance. Anaerobic exercise is high-intensity exercise that lasts for a short period of time and requires muscles to generate explosive force. Energy in anaerobic exercise is primarily generated by the breakdown of glycogen through glycolysis. In some embodiments, anaerobic exercise may include, but is not limited to, sprint running and / or weight training. However, in practice, both aerobic and anaerobic exercise obtain energy through aerobic respiration and glycolysis. In some embodiments, "prolonged exercise" described herein refers to sustained aerobic or anaerobic exercise until muscles need to produce energy from proteins and / or amino acids.

[0029] Experiments have shown that subjects administered with extracts of Phellinus linteus (GKPl) showed increased exercise duration and increased glycogen content in the liver and / or muscles compared to control subjects not administered with extracts of Phellinus linteus (GKPl), indicating that extracts of Phellinus linteus (GKPl) can reliably enhance exercise performance. In the above examples, subjects underwent exercise training for 1 to 10 days or longer periods. Furthermore, subjects administered with extracts of Phellinus linteus (GKPl) showed reduced blood lactic acid and / or urea nitrogen content after exercise compared to control subjects not administered with extracts of Phellinus linteus (GKPl), indicating that extracts of Phellinus linteus (GKPl) can reliably alleviate exercise fatigue.

[0030] The mycelium of Phellinus linteus GKPl (also referred to as strain GKPl) was deposited on July 18, 2019, with the Bioresource Collection and Research Center (BCRC) of the Food Industry Research and Development Institute (FIRDI), Taiwan, at No. 331, Food Road, East District, Hsinchu City, Taiwan 30062, under accession number BCRC 930210. Mycelium of strain GKPl was also deposited on November 12, 2020, with the Patent Microorganisms Depositary of the National Institute of Technology and Evaluation (NITE), Japan, at Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818, under accession number NITE BP-03321. It is further explained that in order to achieve the effects of improving exercise performance and / or reducing exercise fatigue, it is necessary to select a specific strain of Phellinus linteus. If the Phellinus linteus is not the GKP1 strain, it is not possible to expect the effects of improving exercise performance or reducing exercise fatigue.

[0031] The extract of the GKP1 strain may be subjected to a multi-step culture process to increase biomass. The multi-step culture process may be performed using known culture methods. In some embodiments, the multi-step culture process may optionally include a solid culture step, a liquid culture step, and a fermentation step.

[0032] Specifically, to obtain the first culture, the solid culture step may be performed on an extract of the GKP1 strain in a solid medium at 15°C to 30°C for 7 to 14 days. The components of the solid medium are not particularly limited and may be any known solid medium. In some embodiments, the solid medium may include, but is not limited to, a carbon source, a nitrogen source, and potato dextrose agar (PDA).

[0033] To obtain the second culture, the liquid culture step may be performed for 3 to 21 days for the first culture in a liquid medium at 15°C to 30°C, pH 2 to 6, and an agitation speed of 110 rpm to 130 rpm. The components of the liquid medium are not particularly limited and may be adjusted as needed. In some embodiments, the liquid medium may contain, but is not limited to, 1.00% to 3.00% by weight of a comprehensive carbon and nitrogen source, 1.00% to 4.00% by weight of sugars, 0.10% to 1.00% by weight of yeast extract, 0.10% to 1.00% by weight of peptone, 0.01% to 0.50% by weight of inorganic salts, and the balanced amount of water. In some embodiments, the comprehensive carbon and nitrogen source may be, for example, a grain (e.g., wheat flour) and / or a legume (e.g., soybean flour, mung bean flour, and / or cassia flour). In some embodiments, the sugars may include, but are not limited to, monosaccharides (e.g., glucose and / or fructose) and / or disaccharides (e.g., maltose and / or sucrose). In some embodiments, the inorganic salts may be, for example, phosphates (e.g., dipotassium hydrogen phosphate, potassium dihydrogen phosphate) and / or sulfates (e.g., magnesium sulfate and / or ferrous sulfate).

[0034] To obtain a fermentation product of the GKP1 strain containing GKP1 mycelia with increased biomass and growth activity (hereinafter abbreviated as GKP1 fermentation product), the fermentation step may be carried out for 3 to 21 days on the second culture at 15°C to 30°C, pH 2 to 6, and with an agitation rate of 50 rpm to 150 rpm in the fermentation medium. In some embodiments, the fermentation step may be carried out in a fermenter. In some embodiments, the tank pressure of the fermenter is 0.5 kg / cm. 2 ~1.0kg / cm 2 The aeration rate of the fermenter may be 1.0 vvm to 1.5 vvm, where vvm is the volume of air introduced / volume of fermentation medium / minute. The components of the fermentation medium are not particularly limited. In some embodiments, the components of the fermentation medium are the same as those of the liquid medium.

[0035] In some embodiments, the GKPl fermentation product may be optionally dried after the fermentation step to obtain a dried product of the GKPl mycelium (hereinafter referred to as a GKPl dried product). The drying may be performed by a known drying method such as freeze-drying, vacuum drying, or spray drying. In some embodiments, the volume weight ratio (unit: L:kg) of the GKPl fermentation product to the GKPl dried product may be, for example, 100:5 to 100:1.

[0036] The extraction process may be carried out using, for example, a known extraction method. In some embodiments, the GKPl extract may include, but is not limited to, a GKPl hot water extract and / or a GKPl ethanol extract. In some embodiments, the GKPl hot water extract may be obtained by subjecting a GKPl fermentation product and / or a GKPl dried product to a hot water extraction process. In some embodiments, the temperature of the hot water used in the hot water extraction process may be, for example, 80°C to 120°C. In some embodiments, the time period for the hot water extraction process may be, for example, 20 minutes to 40 minutes. In some embodiments, the volume ratio of the GKPl fermentation product and / or the GKPl dried product to the hot water may be, for example, 1 / 50 to 1 / 1, or 1 / 30 to 1 / 10, or 1 / 20.

[0037] In some embodiments, the GKPl ethanol extract may be obtained by subjecting a GKPl fermentation product and / or a dried GKPl product to an ethanol extraction treatment. In some embodiments, the ethanol extraction time may be, for example, 50 to 70 minutes. In some embodiments, the weight ratio of the GKPl fermentation product and / or the dried GKPl product to ethanol may be, for example, 1 / 50 to 1 / 1, or 1 / 30 to 1 / 10, or 1 / 20. In some specific examples, the ethanol extraction treatment may optionally include an ultrasonic step. In some embodiments, the GKPl hot water extract and / or the GKPl ethanol extract may further undergo a drying step to obtain a GKPl fermentation dried product.

[0038] It should be noted that the mycelium of Phellinus linteus GKPl, its fermented product and its dried product all contain active substances that improve exercise performance and reduce exercise fatigue, and the extraction process can only increase the concentration of these active substances, and is not intended to limit the present invention. In other words, the fermented product of the mycelium of Phellinus linteus GKPl and its dried product of the present invention should also have the effect of improving exercise performance and reducing exercise fatigue.

[0039] It should be noted that the extract of Phellinus linteus GKPl described in the present invention has not undergone further purification and therefore contains multiple unknown and inseparable components, among which may be active substances that improve exercise performance and alleviate exercise fatigue. Furthermore, although the present invention demonstrates that the extract of Phellinus linteus GKPl can achieve the effects of improving exercise performance and alleviating exercise fatigue, the extract of Phellinus linteus GKPl contains multiple active substances, and it cannot be reasonably expected that a single substance among them, and / or an extract excluding the unknown and inseparable components, can still achieve the effects of improving exercise performance and alleviating exercise fatigue. Therefore, the extract of Phellinus linteus GKPl described in the present invention cannot be defined by components, properties, or parameters, i.e., it may be impossible or impractical to define the present invention by components, properties, or parameters.

[0040] The extract of the GKPl strain has the effect of improving exercise performance and / or reducing exercise fatigue, and therefore may be used in compositions for improving exercise performance and / or reducing exercise fatigue. In some embodiments, the composition may contain, but is not limited to, a hot water extract and / or an ethanol extract of the GKPl strain as an active ingredient. In another embodiment, the active ingredient of the composition consists of a hot water extract and an ethanol extract of the GKPl strain. In some embodiments, the weight ratio of the hot water extract and the ethanol extract is not particularly limited. In one specific example, the weight ratio of the hot water extract and the ethanol extract is 5:1 to 1:5, or 3:1 to 1:3, or 1:1.

[0041] The composition for improving exercise performance and / or reducing exercise fatigue may be, for example, an oral composition. In some embodiments, the composition may be, for example, a pharmaceutical composition or a food composition. Suitable examples of pharmaceutical compositions include, but are not limited to, pharmaceuticals. Suitable examples of food compositions include, but are not limited to, general foods, health foods, beverages, nutritional supplements, exercise supplements, dairy products, and feed. In some embodiments, the oral composition may optionally contain pharmaceutically and food-acceptable carriers, excipients, and / or additives. In another example, the oral composition may be in the form of, but is not limited to, a powder, tablet, granules, suppository, microcapsule, ampoule, liquid spray, or embolic agent. In some embodiments, the composition comprises an active ingredient (comprising a hot water extract and an ethanol extract of the GKPl strain) and other pharmaceutically and food-acceptable carriers, excipients, and / or additives.

[0042] When administered, an oral composition containing an extract of the GKPl strain may be continuously administered to a subject for a certain period of time. The effective dosage and administration period vary depending on the subject and are not particularly limited. For example, if the subject is an adult, the effective dosage of the extract of the GKPl strain may be, for example, 0.1 g / 60 kg body weight (bw) / day to 2.0 g / 60 kg bw / day, preferably 0.5 g / 60 kg bw / day to 1.5 g / 60 kg bw / day, and more preferably 1.0 g / 60 kg bw / day. In another example, if the subject is a mouse, the effective dosage of the extract of the GKPl strain may be, for example, 0.1 g / kg bw / day to 0.5 g / kg bw / day, preferably 0.1 g / kg bw / day to 0.3 g / kg bw / day, and more preferably 0.15 g / kg bw / day to 0.25 g / kg bw / day. In another example, the administration period may be, for example, at least 28 consecutive days, or may be longer or shorter.

[0043] The following examples are provided to illustrate the application of the present invention, but are not intended to limit the scope of the present invention. Those skilled in the art may make various changes and modifications without departing from the spirit and scope of the present invention. Example 1: Preparation of extract of GKPl 1.1 Origin of the Phellinus linteus strain GKPl

[0044] The accession number of the mycelium of the Phellinus linteus strain GKP1 used in this example is BCRC 930210 or NITE BP-03321. The strain GKP1 is a mycelium isolated from the fruiting body of wild Phellinus linteus in China. For the microbiological properties and cultivation method of Phellinus linteus GKP1, please refer to Taiwan Patent (Publication No.: TW I729928 B), Taiwan Patent (Publication No.: TW I766394 B), and Japanese Patent Application (Publication No.: JP 2022067043 A), which are incorporated herein by reference. 1.2 Preparation of extract of Phellinus linteus strain GKPl

[0045] The GKPl mycelium was inoculated into potato dextrose agar medium and cultured solid for 7 days at 25°C to obtain a first culture. The first culture was then cultured in a liquid medium at 25°C, pH 4.5, and a shaking speed of 120 rpm for 14 days to obtain a second culture. The liquid medium contained 1.0% by weight of a comprehensive carbon and nitrogen source, 1.0% by weight of sugars, 0.3% by weight of yeast extract, 0.3% by weight of peptone, and 0.1% by weight of inorganic salts. The specific components of the comprehensive carbon and nitrogen source, sugars, and inorganic salts are well known to those skilled in the art and can be adjusted as needed; therefore, further explanation is omitted here.

[0046] Next, at 25°C, pH 5, vibration speed of 80 rpm, and 0.5 kg / cm 2 The second culture was subjected to a 14-day fermentation step using the above culture medium at a tank pressure of 1000 kJ / L and an air aeration rate of 1.0 vvm, thereby obtaining a fermented product of the GKPl mycelium. The fermented product of the GKPl mycelium was then freeze-dried to obtain a freeze-dried powder of the fermented product of the GKPl mycelium (hereinafter referred to as GKPl freeze-dried powder). In this example, 3 kg of GKPl freeze-dried powder could be obtained from 100 L of the fermented product of the GKPl mycelium.

[0047] Next, a hot water extract and an ethanol extract of the GKPl strain (hereinafter referred to as the GKPl hot water extract and the GKPl ethanol extract, respectively) were prepared. The GKPl hot water extract was obtained by adding the GKPl freeze-dried powder to distilled water and then subjecting it to hot water extraction at 100°C for 30 minutes, where the volume ratio of the GKPl freeze-dried powder to distilled water was 1 / 20. The GKPl ethanol extract was obtained by adding the GKPl freeze-dried powder to ethanol and then subjecting it to ethanol extraction at 25°C for 60 minutes and an ultrasonic step, where the weight ratio of the GKPl freeze-dried powder to ethanol was 1 / 20. The GKPl hot water extract and the GKPl ethanol extract were mixed in a 1:1 weight ratio to obtain the GKPl strain extract (hereinafter referred to as the GKPl extract). The GKPl strain extract was further subjected to a freeze-drying step to obtain the GKPl freeze-dried powder. Example 2: Evaluation of the effect of GKPl extract in improving exercise performance and reducing exercise fatigue 2.1 Experimental animals

[0048] In the following example, 12 6-week-old ICR male mice were divided into a control group and a Phellinus linteus group, with 6 mice in each group. From days 1 to 28, the mice were orally administered water (control group) or GKPl extract (experimental group) twice daily at 9:00 AM. The mice's weights, food intake, and water intake were observed and recorded. From days 29 to 37, a weighted swimming exhaustion experiment (unit: minutes) and a treadmill exhaustion experiment (unit: minutes) were conducted. The change ratio of blood lactate content before and after short-term swimming and after rest, urea nitrogen content (unit: mg / dL) after long-term swimming and after rest, and glycogen content in the liver and muscle (mg / g liver or mg / g muscle) were measured. 2.2 Weighted swimming exhaustion experiment

[0049] On the 31st day, a weighted swimming exhaustion experiment was conducted. Prior to the experiment, the mice were fasted for 12 hours. Thirty minutes before the experiment, the mice were given water (control group) and GKPl extract (experimental group) twice. During the experiment, a lead sheet weighing 5% of the mouse's body weight was fastened to the base of the mouse's tail, and the mouse was placed in a tank and forced to swim. The water temperature in the tank was maintained at 27±1°C, and the experiment was conducted with individual swimming to avoid any influence on the experimental results due to pushing during group swimming.

[0050] The mouse's limbs must be kept moving throughout the entire experiment. If the mouse floats in the water and does not move its limbs, a stirring rod is used nearby to stir it. The experiment is terminated when the mouse loses coordination and is unable to return to the surface within 7 seconds (i.e., swims to exhaustion). The time from the start to the end of the experiment is calculated as the mouse's weight-applied swimming exhaustion time (i.e., duration). The experimental results are recorded in Table 1.

[0051] [Table 1]

[0052] Please refer to Figure 1, which is a bar graph of the weighted swimming exhaustion time of mice in the control group and the experimental group shown in Table 1, where the horizontal axis represents the group and the vertical axis represents the weighted swimming exhaustion time (unit: minutes), and the figure number "***" indicates a statistically significant difference (p<0.001) compared to the control group.

[0053] As shown in Figure 1, the weighted swimming exhaustion time of the mice in the experimental group was significantly longer than that of the control group (an increase of approximately 78.89%), which indicates that GKPl extract can reliably increase the exercise duration of the mice and has the effect of improving their exercise performance. 2.3 Treadmill exhaustion experiment

[0054] On the 31st day, a treadmill exhaustion experiment was conducted. Prior to the experiment, the mice were fasted for 12 hours. Thirty minutes before the experiment, the mice were administered water (control group) and GKPl extract (experimental group) twice. During the experiment, the mice were placed on the treadmill belt, which had a 5° gradient, with the lower end being the lower end. An electric shock grid was installed at the end of the treadmill. When the mouse stopped running and a certain part of its body came into contact with the electric shock grid, the mouse was shocked. At the beginning of the experiment, the treadmill speed was set to 10 m / min. After the experiment lasted for 5 minutes, the speed was increased by 2 m / min. The experiment was terminated when the mouse exhausted. The criteria for mouse exhaustion were that the mouse received multiple electric shocks or received electric shocks for 5 consecutive seconds. The experimental results are recorded in Table 2.

[0055] [Table 2]

[0056] Referring to Figure 2, which is a bar graph of the treadmill exhaustion times of mice in the control and experimental groups shown in Table 2, where the horizontal axis represents the group and the vertical axis represents the treadmill exhaustion time (unit: minutes), and the figure number "***" indicates a statistically significant difference (p<0.001) compared to the control group.

[0057] As shown in Figure 2, the treadmill exhaustion time of the mice in the experimental group was significantly longer than that of the control group (an increase of approximately 62.42%), indicating that GKPl extract can reliably increase the exercise duration of the mice and has the effect of improving their exercise performance. 2.4 Blood lactate increase rate and blood lactate removal rate after rest

[0058] On day 35, the increase in blood lactate after a short exercise session and the removal of lactate after a rest session were measured. The mice were first administered water (control group) and GKPl extract (experimental group) twice. After 30 minutes, the first blood sample was taken to obtain a blood sample before swimming. The mice were then allowed to swim for 10 minutes in a 30°C water bath. A second blood sample was taken to obtain a blood sample after swimming. After a 20-minute rest, a third blood sample was taken to obtain a blood sample after swimming and resting.

[0059] The lactate content in blood samples taken before swimming, after swimming, and after swimming and resting was measured to obtain the lactate content in blood before swimming, after swimming, and after swimming and resting. The lactate content in blood was measured using a fully automated biochemistry analyzer (name: HITACHI 7060; manufacturer: Hitachi, Tokyo, Japan).

[0060] The lactate increase rate in the blood after swimming was calculated by calculating the ratio of the difference between the lactate content in the blood after swimming and the lactate content in the blood before swimming to the lactate content in the blood before swimming.The lactate removal rate in the blood after resting was calculated by calculating the ratio of the difference between the lactate content in the blood after swimming and the lactate content in the blood after swimming and resting to the lactate content in the blood after swimming.The experimental results are recorded in Table 3.

[0061] [Table 3]

[0062] Referring to Figures 3A and 3B, these are bar graphs of the blood lactate increase rate (Figure 3A) and removal rate (Figure 3B) of mice in the control and experimental groups shown in Table 3, respectively, where the horizontal axis represents the group, and the vertical axis represents the blood lactate increase rate (Figure 3A, no units) and blood lactate removal rate (Figure 3B, no units), respectively, and the figure number "*" indicates a statistically significant difference (p<0.05) compared to the control group.

[0063] As shown in Figure 3A, the blood lactate increase rate of mice in the experimental group was significantly lower than that of the control group, indicating that GKPl extract can reduce blood lactate accumulation during exercise.Next, as shown in Figure 3B, the blood lactate removal rate of mice in the experimental group after rest was significantly higher than that of the control group, indicating that GKPl extract can improve lactate removal during rest and effectively alleviate exercise fatigue. 2.5 Blood urea nitrogen content before and after long-term swimming

[0064] On day 37, the blood urea nitrogen content of the mice was measured before and after the long-distance swimming. The mice were first administered water (control group) and GKPl extract (experimental group) twice, and 30 minutes later, blood was collected to obtain a blood sample before the long-distance swimming. The mice were then allowed to swim in a water tank at 30°C for 60 minutes, after which blood was collected again to obtain a blood sample after the long-distance swimming.

[0065] The urea nitrogen content of the blood samples taken before and after the long swim was measured to obtain the urea nitrogen content of the blood before and after the long swim. The experimental results are recorded in Table 4.

[0066] [Table 4]

[0067] Referring to Figure 4, it is a bar graph of the urea nitrogen content in the blood of mice in the control group and the experimental group shown in Table 4 before and after long-distance swimming, where the horizontal axis represents, from left to right, before and after long-distance swimming, and the vertical axis represents the urea nitrogen content in the blood (unit: mg / dL), with the straight bars 410 and 420 representing the control group and the experimental group, respectively, and the figure number "*" indicates a statistically significant difference (p<0.05) compared to the control group.

[0068] As shown in Figure 4, there was no statistically significant difference in the urea nitrogen content in the blood of mice in the experimental and control groups before long-term swimming (i.e., during normal activity), indicating that the GKPl extract does not affect the protein and / or amino acid metabolism of mice during normal activity. However, after long-term swimming, the urea nitrogen content in the blood of mice in the experimental group was significantly lower than that of the control group, indicating that the GKPl extract can reliably reduce the accumulation of urea nitrogen and has the effect of effectively alleviating exercise fatigue. 2.6 Glycogen content in liver and muscle

[0069] On day 39, the glycogen content in the livers and muscles of the mice was analyzed. First, the mice were administered water (control group) and GKPl extract (experimental group) twice, and then sacrificed 30 minutes later. The livers and hind leg crural muscles of the mice were removed, washed, dried, and weighed. Next, tissues from the same locations on the liver and hind leg crural muscles were removed and homogenized using a tissue homogenizer (Bullet Blender®; manufacturer: Next Advance, Cambridge, Massachusetts, USA) with 10% glacial perchloric acid (5 times the tissue volume) to obtain a tissue homogenate.

[0070] The tissue homogenate was centrifuged at 12,000 x g for 15 minutes at 4°C to obtain the supernatant. 30 μL of the supernatant was mixed with 200 μL of iodine-potassium iodide reagent for 10 minutes to react the glycogen in the supernatant with the iodine-potassium iodide, producing a brown substance. The absorbance of the supernatant was measured at 460 nm using an ELISA (Tecan Infinite M200; manufacturer: Tecan Austria, Salzburg, Austria). After creating a standard curve using standard glycogen (purchased from Sigma), the glycogen content (mg / g liver or mg / g muscle) in the liver and muscle tissues of each group of mice was calculated. The results are recorded in Table 5.

[0071] [Table 5]

[0072] Referring to Figure 5, which is a bar graph of glycogen content in the liver and muscle of mice in the control and experimental groups shown in Table 5, in which the horizontal axis represents the site, from left to right being liver and muscle, and the vertical axis represents glycogen content (units: mg / g liver, mg / g muscle), bar 510 and bar 520 represent the control group and the experimental group, respectively, and the figure number "*" indicates a statistically significant difference (p<0.05) compared to the control group.

[0073] As shown in Figure 5, after several days of training, the glycogen content in the liver and muscle of the mice in the experimental group was significantly higher than that in the control group, indicating that GKPl extract can reliably increase the glycogen content in the liver and muscle, thereby improving exercise performance.

[0074] Therefore, the above-mentioned specific culture methods, extraction methods, dosage amounts, dosage forms, experimental animals, and evaluation methods are used only to exemplify the compositions for improving exercise performance and reducing exercise fatigue containing the Phellinus linteus GKPl extract of the present invention. However, as will be understood by those skilled in the art, other culture methods, extraction methods, dosage amounts, dosage forms, experimental animals, and evaluation methods may be applied to compositions for improving exercise performance and reducing exercise fatigue without departing from the spirit and scope of the present invention, and are not limited to the above. For example, the multiple culture step treatment is used only to improve the biomass and biological activity of the Phellinus linteus GKPl extract, and the Phellinus linteus GKPl extract obtained by multiple culture step treatments under known conditions should also have the effect of improving exercise performance and reducing exercise fatigue.

[0075] As can be seen from the above examples, the composition for improving exercise performance and reducing exercise fatigue containing the extract of Phellinus linteus GKPl of the present invention has the advantage that by using the extract of a specific strain of Phellinus linteus GKPl, it can improve exercise endurance, glycogen content in the liver and muscles, and reduce the content of lactic acid and urea nitrogen in the blood after exercise, and therefore it can be used as an active ingredient in the composition for improving exercise performance and reducing exercise fatigue.

[0076] While the present invention has been disclosed above in terms of several specific embodiments, it should be understood that various modifications, changes, and substitutions can be made to the above-described invention, and that in some cases some features of the embodiments of the present invention may be employed while other features are not correspondingly used, without departing from the spirit and scope of the present invention. Therefore, the spirit and scope of the present invention and the claims should not be limited to the above-described exemplary embodiments. [Explanation of symbols]

[0077] 410, 420, 510, 520 straight rod [Deposit of biological materials] The Phellinus linteus was deposited at the Biological Resources Center (BCRC), Food Industry Development Research Institute, 331 Food Road, Hsinchu, Taiwan on July 18, 2019, with the accession number BCRC 930210.

Claims

1. A composition for increasing glycogen content in muscle, comprising an extract of Phellinus linteus GKP1, the composition comprising an extract of mycelia of Phellinus linteus GKP1 as an active ingredient, the extract comprising a hot water extract and an ethanol extract, the weight ratio of the hot water extract to the ethanol extract being 5:1 to 1:5, the ethanol extract being obtained by subjecting mycelia of Phellinus linteus GKP1 to ethanol extraction treatment, and the Phellinus linteus GKP1 was approved by the Bioresource Collection and Research Center of the Food Industry Research and Development Institute (FIRDI) in Taiwan on July 18, 2019. The composition for improving muscle glycogen content contains an extract of Phellinus linteus GKP1, which has been deposited at the British Cancer Center (BCRC) under accession number BCRC 930210, thereby improving muscle glycogen content in subjects who do not have muscle atrophy.

2. 2. The composition for increasing glycogen content in muscle containing the extract of Phellinus linteus GKP1 according to claim 1, wherein the hot water extract is obtained by subjecting the mycelium of Phellinus linteus GKP1 to hot water extraction treatment with water at 80°C to 120°C for 20 minutes to 40 minutes.

3. The composition for improving glycogen content in muscle containing the extract of Phellinus linteus GKP1 according to claim 1, wherein the subject has undergone exercise training for 1 to 10 days.

4. The composition for increasing glycogen content in muscle containing the extract of Phellinus linteus GKP1 according to claim 1, wherein the composition is a pharmaceutical composition or a food composition.

5. 2. The composition for increasing glycogen content in muscle containing the extract of Phellinus linteus GKP1 according to claim 1, wherein the composition further comprises a carrier, excipient and / or additive acceptable for pharmaceuticals and / or foods.

6. 2. The composition for improving glycogen content in muscle, comprising the extract of Phellinus linteus GKPl according to claim 1, wherein the exercise duration of the subjects administered with the extract of Phellinus linteus GKPl is increased compared to the control subjects not administered with the extract of Phellinus linteus GKPl, and the control subjects do not have muscle atrophy.

7. 2. A composition for improving glycogen content in muscle, comprising the extract of Phellinus linteus GKP1 according to claim 1, which reduces the urea nitrogen content in blood after exercise in subjects who have been administered the composition for at least 28 consecutive days.

8. 2. The composition for increasing glycogen content in muscle containing the extract of Phellinus linteus GKP1 according to claim 1, wherein the effective dosage is 0.1 g / 60 kg body weight (b.w.) / day to 2.0 g / 60 kg b.w. / day when the subject is an adult.

9. The composition for increasing glycogen content in muscle containing the extract of Phellinus linteus GKP1 according to claim 1, wherein the effective dosage is 0.1 g / kg b.w. / day to 0.5 g / kg b.w. / day when the subject is a mouse.

10. 2. The composition for increasing glycogen content in muscle, comprising the extract of Phellinus linteus GKPl according to claim 1, wherein the subjects administered with the extract of Phellinus linteus GKPl have a lower blood lactic acid content after exercise compared to the control subjects not administered with the extract of Phellinus linteus GKPl, and the control subjects do not have muscle atrophy.

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