Composition for improving muscle quality

Abstract

Provided are technique for improving muscle quality and technique related thereto. The present invention improves muscle quality by using a choline compound such as sn-Glycero-3-phosphocholine (αGPC) or citicoline.

Description

Composition for improving muscle quality

[0001] The present invention relates to a technique for improving muscle quality and related techniques. In particular, the present invention may relate to a technique for slow-twitch muscle formation and related techniques.

[0002] Muscles are composed of bundles of muscle fibers. Muscle fibers are classified into slow-twitch fibers (Type I muscle fibers) and fast-twitch fibers (Type II muscle fibers). Compared to fast-twitch fibers, slow-twitch fibers have lower contractile force and contraction speed, but are less prone to fatigue, contributing to endurance. Therefore, increasing the amount of slow-twitch fibers in muscles (i.e., converting muscles to slow-twitch fibers) is expected to result in health benefits such as improved endurance. Furthermore, slow-twitch fibers have the property of consuming more energy than fast-twitch fibers. Therefore, increasing the amount of slow-twitch fibers in muscles (i.e., converting muscles to slow-twitch fibers) is expected to increase energy consumption, thereby resulting in health benefits such as weight loss.

[0003] A technique for promoting lipid metabolism using sn-glycero-3-phosphocholine (αGPC) has been reported (Patent Document 1).

[0004] A technique for improving the mental state and body composition of mammals using Igirin has been reported (Patent Document 2). This document lists citicoline as an example of an ingredient that may be used in combination with Igirin.

[0005] A technique for improving mitochondrial energy production and its efficiency using an NAD+ precursor and an ATP booster has been reported (Patent Document 3). This document lists nutrients that protect mitochondria, such as citicoline, as examples of ingredients that may be used in combination with the NAD+ precursor and the ATP booster.

[0006] Patent Publication No. 2014-051459US2018-0071273US2014-0134283

[0007] An object of the present invention is to provide a technique for improving muscle quality and related techniques.

[0008] As a result of intensive research to solve the above problems, the present inventors have found that muscle quality can be improved by using choline compounds such as sn-glycero-3-phosphocholine (αGPC) and citicoline, and have completed the present invention.

[0009] That is, the present invention can be exemplified as follows: [1] A composition for improving muscle quality, containing a cholinergic compound. [2] A composition for obtaining an effect based on muscle quality improvement, containing a cholinergic compound. [3] The composition, wherein the effect based on muscle quality improvement is one or more effects selected from the group consisting of an effect of increasing the amount of mitochondria in muscle, an effect of improving body composition, an effect of improving endurance, an effect of improving metabolism, an effect of improving balance ability, an effect of improving muscle strength, an effect of improving walking speed, an effect of toning the body, an effect of improving basic physical fitness, an effect of improving training effects, an effect of improving rehabilitation effects, an effect of regulating the body temperature of a subject, an effect of regulating the blood glucose level of a subject, an effect of regulating the blood lipid concentration of a subject, and an effect of preventing, ameliorating, and / or treating undesirable muscle-related symptoms. [4] The composition, wherein the symptom is one or more symptoms selected from the group consisting of disuse muscle atrophy, neurogenic muscle atrophy, myogenic muscle atrophy, myasthenia gravis, cachexia, frailty, musculoskeletal syndrome, metabolic syndrome, decreased basic physical strength, decreased walking speed, lower back pain, stiff shoulders, headache, hyperglycemia, hyperlipidemia, vascular disorder, hypothermia, and sensitivity to cold. [5] The composition, wherein the improvement of muscle quality is muscle slow-twitch muscle contraction. [6] The composition, wherein the cholinergic compound is one or more components selected from the group consisting of sn-glycero-3-phosphocholine, citicoline, choline, phosphate choline, phosphocholine, phosphatidylcholine, sphingomyelin, and serine. [7] The composition, wherein the cholinergic compound is one or more components selected from the group consisting of sn-glycero-3-phosphocholine, citicoline, and serine. [8] The composition, wherein the composition is a pharmaceutical composition. [9] The composition, wherein the composition is a food composition.

[10] The composition, wherein the content of the cholinergic compound is 100 ppm (w / w) to 99.9% (w / w).

[11] The composition, wherein the composition is used in combination with physical exercise.

[12] The composition, wherein, when used in combination with physical exercise, muscle strength is improved earlier in a subject to which the composition is administered, compared to when the composition is not used in combination with physical exercise.

[13] A method for improving muscle quality in a subject, comprising the step of administering a cholinergic compound to the subject.

[14] A method for obtaining an effect based on muscle quality improvement in a subject, comprising the step of administering a cholinergic compound to the subject.

[15] The method, wherein the effect based on muscle quality improvement is one or more effects selected from the group consisting of an effect of increasing the amount of mitochondria in muscle, an effect of improving body composition, an effect of improving endurance, an effect of improving metabolism, an effect of improving balance ability, an effect of improving muscle strength, an effect of improving walking speed, an effect of toning the body, an effect of improving basic physical fitness, an effect of improving training effects, an effect of improving rehabilitation effects, an effect of regulating the body temperature of a subject, an effect of regulating the blood glucose level of a subject, an effect of regulating the blood lipid concentration of a subject, and an effect of preventing, ameliorating, and / or treating an undesirable symptom related to muscle.

[16] The method, wherein the symptom is one or more symptoms selected from the group consisting of disuse muscle atrophy, neurogenic muscle atrophy, myogenic muscle atrophy, myasthenia gravis, cachexia, frailty, musculoskeletal syndrome, metabolic syndrome, decreased basic physical strength, decreased walking speed, lower back pain, stiff shoulders, headache, hyperglycemia, hyperlipidemia, vascular disorder, hypothermia, and sensitivity to cold.

[17] The method, wherein the dose of the cholinergic compound is 0.01 mg / kg body weight / day or more.

[18] The method, wherein the dose of the cholinergic compound is 1000 mg / kg body weight / day or less.

[19] The method, wherein the method is used in conjunction with physical exercise.

[20] The method, wherein, when used in conjunction with physical exercise, muscle strength improves earlier in the subject compared to when not used in conjunction with physical exercise.

[21] A method for improving muscle quality in cultured meat, comprising a step of culturing animal cells in the presence of a cholinergic compound.

[22] A method for producing cultured meat, comprising a step of culturing animal cells in the presence of a choline-based compound.

[23] The method, wherein the cultured meat produced is cultured meat with improved muscle quality.

[24] The method, wherein the improvement in muscle quality is conversion to slow-twitch muscle.

[25] The method, wherein the conversion to slow-twitch muscle improves the texture and / or flavor of the cultured meat.

[26] The method, wherein the improvement in texture is improvement in the water retention capacity and / or juiciness of the cultured meat.

[27] The method described above, wherein the improved flavor is an improvement in the taste of the cultured meat due to an increase in the amount of free amino acids.

[28] The method described above, wherein the choline-based compound is one or more components selected from the group consisting of sn-glycero-3-phosphocholine, citicoline, choline, phosphate choline, phosphocholine, phosphatidylcholine, sphingomyelin, and serine.

[29] The method described above, wherein the choline-based compound is one or more components selected from the group consisting of sn-glycero-3-phosphocholine, citicoline, and serine.

[0010] Figure showing changes in Myh7 gene expression levels with citicoline administration. The Myh7 gene expression levels were expressed as relative values ​​(mean ± standard deviation) with the control group's Myh7 gene expression level set to 1 for 24, 48, and 72 hours after citicoline administration. Significance tests were performed using a two-group test (*: p<0.05). Figure showing changes in Myh7 gene expression levels with αGPC administration. The Myh7 gene expression levels were expressed as relative values ​​(mean ± standard deviation) with the control group's Myh7 gene expression level set to 1 for 24, 48, and 72 hours after citicoline administration. Significance tests were performed using a two-group test (*: p<0.05). Photograph of an animal grip strength tester. Figure showing the muscle strength of mice 2 days after the start of feeding the donor diet. Results are expressed as mean ± standard deviation. Significance tests were performed using a Dunnett's test (*: p<0.05). This figure shows the muscle strength of mice 8 days after the start of donor diet feeding. Results are shown as mean ± standard deviation. Significance was tested using Dunnett's test (*: p<0.05). This figure shows the change in Myh7 gene expression level due to serine supplementation. The Myh7 gene expression level 24 hours after serine supplementation is shown as a relative value (mean ± standard deviation), with the Myh7 gene expression level in the control group set at 1. Significance was tested using Dunnett's test (*: p<0.05). This figure shows the change in fat weight per body weight of mice after the start of donor diet feeding. Results are shown as mean values. This figure shows the fat weight per body weight of mice 26 days after the start of donor diet feeding. Results are shown as mean ± standard deviation. A multiple comparison test against the control group (Group 2) was performed using Dunnett's test to test for significance (*: p<0.05). This figure shows the change in muscle weight per body weight of mice after the start of donor diet feeding. Results are shown as mean values. Figure showing muscle weight per body weight of mice on day 26 after the start of feeding the donated feed. Results are shown as mean values ​​± standard deviation. To test for significance, a multiple comparison test was performed against the control group (Group 2) using Dunnett's test (*: p<0.05). Figure showing citrate synthase (CS) activity in the gastrocnemius muscle of mice on day 26 after the start of feeding the donated feed. Results are shown as mean values ​​± standard deviation.To test for significance, a multiple comparison test was performed using Dunnett's test against the control group (Group 2). Figure showing the change in muscle strength over time in mice after the start of feeding the donor diet. Results are shown as the mean. Figure showing the muscle strength of mice on day 4 after the start of feeding the donor diet. Results are shown as the mean ± standard deviation. To test for significance, a multiple comparison test was performed using Dunnett's test against the control group (Group 2) (**: p<0.01). Figure showing the muscle strength of mice on day 25 after the start of feeding the donor diet. Results are shown as the mean ± standard deviation. To test for significance, a multiple comparison test was performed using Dunnett's test against the control group (Group 2) (**: p<0.01).

[0011] The present invention will be described in detail below.

[0012] <1> Active ingredient In the present invention, a cholinergic compound is used as an active ingredient. The cholinergic compound is also referred to as an "active ingredient."

[0013] By utilizing an active ingredient, muscle quality can be improved, i.e., a muscle quality improving effect can be obtained. This effect is also referred to as a "muscle quality improving effect." Use of an active ingredient can be achieved by administering the active ingredient to a subject or by culturing animal cells in the presence of the active ingredient. That is, for example, by administering the active ingredient to a subject, muscle quality in the subject can be improved, i.e., a muscle quality improving effect can be obtained. Furthermore, for example, by culturing animal cells in the presence of the active ingredient, muscle quality can be improved in muscle tissue formed by the cultured cells (e.g., cultured meat), i.e., a muscle quality improving effect can be obtained. The term "improving muscle quality in cultured meat" can be used interchangeably or equivalently with the terms "improving the meat quality of cultured meat" or "controlling the meat quality of cultured meat." Examples of muscles include skeletal muscles. Examples of skeletal muscles include trunk muscles (i.e., skeletal muscles of the trunk) and limb muscles (i.e., skeletal muscles of the upper and lower limbs). Examples of skeletal muscles include, in particular, skeletal muscles involved in maintaining posture. Examples of skeletal muscles involved in maintaining posture include trunk muscles such as the cervical flexors, cervical extensors, erector spinae, abdominal muscles, and iliopsoas, as well as lower limb skeletal muscles such as the gluteus maximus, quadriceps femoris, biceps femoris, semitendinosus, semimembranosus, tibialis anterior, gastrocnemius, and soleus. The muscle quality improving effect may be obtained in one muscle, or in two or more muscles. For example, the muscle quality improving effect may be obtained in a region with a high proportion of slow-twitch muscles, a region with a high proportion of fast-twitch muscles, or both.

[0014] An example of an improvement in muscle quality is the conversion of muscle to slow-twitch muscle. That is, an example of the muscle quality improvement effect is the conversion of muscle to slow-twitch muscle. The effect of converting muscle to slow-twitch muscle is also referred to as the "slow-twitch conversion effect." "Conversion of muscle to slow-twitch muscle" means an increase in the amount of slow-twitch muscle in muscle. "Amount of slow-twitch muscle in muscle" may mean the absolute amount of slow-twitch muscle in muscle, the relative amount of slow-twitch muscle in muscle (i.e., the ratio of the amount of slow-twitch muscle to the muscle mass), or both. That is, the use of an active ingredient may increase the absolute amount and / or relative amount of slow-twitch muscle in muscle. The use of an active ingredient may at least increase the relative amount of slow-twitch muscle in muscle. The total muscle mass may increase with an increase in the amount of slow-twitch muscle in muscle. The amount of slow-twitch muscle includes the number of slow-twitch muscle fibers, the cross-sectional area of ​​slow-twitch muscle fibers, and the volume of slow-twitch muscle fibers. That is, for example, the relative amount of slow muscle fibers in a muscle includes the ratio of the number of slow muscle fibers to the number of muscle fibers in the muscle, the ratio of the cross-sectional area of ​​slow muscle fibers to the cross-sectional area of ​​the muscle, and the ratio of the volume of slow muscle fibers to the muscle volume. "Slow muscle fiber" refers to Type I muscle fiber. The use of an active ingredient may increase, for example, one or more of the parameters considered to be the amount of slow muscle fibers, as exemplified above. The use of an active ingredient may increase, decrease, or remain unchanged the amount of fast muscle fibers in the muscle. "The amount of fast muscle fibers in a muscle" may refer to the absolute amount of fast muscle fibers in the muscle, the relative amount of fast muscle fibers in the muscle (i.e., the ratio of the amount of fast muscle fibers to the amount of muscle), or both. In one aspect, the use of an active ingredient may increase the absolute amount of fast muscle fibers in the muscle in addition to increasing the absolute and / or relative amount of slow muscle fibers in the muscle. In one aspect, the use of an active ingredient may increase the absolute amount of fast muscle fibers in the muscle in addition to increasing the absolute amount of slow muscle fibers in the muscle. In one embodiment, the use of an active ingredient may decrease the relative amount of fast-twitch muscle fibers in a muscle while increasing the relative amount of slow-twitch muscle fibers in the muscle. The amount of fast-twitch muscle fibers includes the number of fast-twitch muscle fibers, the cross-sectional area of ​​fast-twitch muscle fibers, and the volume of fast-twitch muscle fibers. "Fast-twitch muscle fibers" refers to Type II muscle fibers. Type II muscle fibers include Type IIa muscle fibers, Type IIb muscle fibers, and Type IIx muscle fibers.

[0015] Improvements in muscle quality include improvements in meat texture and meat flavor. That is, muscle quality improvement effects also include improvements in meat texture and meat flavor. Improvements in meat texture and meat flavor may both be obtained, for example, in edible meat (e.g., meat from animals for meat production or cultured meat for consumption). Improvements in meat texture and meat flavor may both be achieved, for example, by converting muscle into slow-twitch muscle. Improvements in meat texture include improvements in meat water retention and juiciness. Improvements in meat flavor include improvements in meat taste due to increased amounts of free amino acids. For example, it has been reported that the proportion of slow-twitch muscle in pork shows a positive correlation with the desirable flavor of pork (Y.K. Kang, et al., Meat Sci., 89, 384 (2011); A. Karlsson, et al., J. Anim. Sci., 71, 930 (1993)). It has also been reported that increasing the ratio of slow-twitch muscle fibers in pork improves its water retention and juiciness (YK Kang, et al., Meat Sci., 89, 384 (2011); GD Kim, et al., J. Anim. Sci., 91, 5525 (2013)). It has also been reported that slow-twitch muscle fibers contain more free amino acids than fast-twitch muscle fibers, which affect the flavor of beef (D. Mashima, et al., Anim. Sci. J., 90, 604 (2019)).

[0016] The muscle quality improving effect can be confirmed by comparing muscle quality when the active ingredient is not used with that when the active ingredient is used. For example, the slow-twitch muscle-improving effect can be confirmed by comparing the amount of slow-twitch muscle in the muscle when the active ingredient is not used with that when the active ingredient is used. That is, if muscle quality is improved when the active ingredient is used compared to when the active ingredient is not used (e.g., the amount of slow-twitch muscle in the muscle is greater), it can be determined that a muscle quality improving effect (e.g., a slow-twitch muscle-improving effect) has been achieved. "When the active ingredient is used" may mean, for example, a time point after the active ingredient is used in the manner described in the method of the present invention below. "When the active ingredient is not used" may simply mean, for example, a time point before the active ingredient is used. That is, for example, if muscle quality is improved after the active ingredient is used compared to before the active ingredient is used (e.g., the amount of slow-twitch muscle in the muscle is greater), it can be determined that a muscle quality improving effect (e.g., a slow-twitch muscle-improving effect) has been achieved. Furthermore, "when the active ingredient is not used" may more accurately mean, for example, a time point after a period of time has passed without the active ingredient being used (e.g., the same period as the period during which the active ingredient was used). Therefore, for example, in cases where it is assumed that muscle quality will deteriorate over time (e.g., the amount of slow-twitch muscle in the muscle will decrease), "improving muscle quality through the use of an active ingredient" (e.g., "the use of an active ingredient converts the muscle to slow-twitch muscle") means that the deterioration of muscle quality (e.g., the decrease in the amount of slow-twitch muscle in the muscle) is less than expected due to the use of the active ingredient, and does not require that muscle quality be improved after the use of the active ingredient compared to before the use of the active ingredient (e.g., the amount of slow-twitch muscle in the muscle is greater). In other words, "improving muscle quality" (e.g., "converting the muscle to slow-twitch muscle") also includes a reduction in the deterioration of muscle quality (e.g., the decrease in the amount of slow-twitch muscle in the muscle). Examples of cases where muscle quality is assumed to deteriorate over time (e.g., the amount of slow-twitch muscle in the muscle) include when the subject has a factor that can cause a deterioration of muscle quality (e.g., a decrease in the amount of slow-twitch muscle in the muscle), such as disease or aging. When an active ingredient is administered to a subject for use, the condition of the subject after a period of time has passed without the use of the active ingredient (e.g., muscle quality and the amount of slow-twitch muscle in the muscle) can be inferred by taking into account various conditions such as the age and health condition of the subject.When an active ingredient is used in the culture of animal cells, the state of muscle tissue formed by the cultured cells after a period of time without the active ingredient (e.g., muscle quality and the amount of slow-twitch muscle in the muscle) can be confirmed, for example, by culturing the animal cells in the absence of the active ingredient. The amount of slow-twitch muscle can be measured, for example, by known methods. Methods for measuring the amount of slow-twitch muscle include muscle biopsy, magnetic resonance imaging (MRI), and field tests. The amount of slow-twitch muscle in the muscle when the active ingredient is used may be, for example, 101% or more, 103% or more, 105% or more, 110% or more, 115% or more, 120% or more, 125% or more, or 130% or more, assuming the amount of slow-twitch muscle in the muscle without the active ingredient is 100%. Furthermore, the slow-twitch muscle-forming effect can be confirmed, for example, using the expression of the Myh7 gene in the muscle as an indicator. The Myh7 gene is a gene highly expressed in slow-twitch muscle fibers. In other words, it can be determined that the slow-twitch muscle-promoting effect has been achieved when the expression level of the Myh7 gene in the muscle is higher compared to when the active ingredient is not used. Furthermore, the effects of improving the texture and flavor of meat can both be confirmed, for example, by sensory evaluation by a specialist panel.

[0017] Furthermore, by utilizing the active ingredient, an effect based on improvement of muscle quality (e.g., slow-twitch muscle) may be obtained. For example, by administering the active ingredient to a subject, an effect based on improvement of muscle quality (e.g., slow-twitch muscle) may be obtained in the subject.

[0018] It is expected that improving muscle quality (e.g., slow-twitch muscle) can increase the amount of mitochondria in muscle. In other words, an effect based on improving muscle quality (e.g., slow-twitch muscle) includes increasing the amount of mitochondria in muscle. The amount of mitochondria in muscle can be measured, for example, by measuring the amount of mitochondrial DNA in muscle or the activity of citrate synthase (CS) in muscle. For example, an increase in the amount of mitochondrial DNA in muscle or CS activity in muscle can be considered to indicate an increase in the amount of mitochondria in muscle. CS activity can be measured using a CS activity measurement kit, such as the Citrate Synthase Assay Kit ab239712 (Abcam).

[0019] Improving muscle quality (e.g., slow-twitch muscle conversion) is expected to improve the body composition of a subject. Improving muscle quality (e.g., slow-twitch muscle conversion) is also expected to improve the endurance of a subject. Improving muscle quality (e.g., slow-twitch muscle conversion) is also expected to improve the metabolism of a subject, such as basal metabolism. Specifically, for example, if the amount of mitochondria in muscle increases with an increase in the amount of slow-twitch muscle, it is expected to improve the endurance and / or metabolism of a subject. Improving muscle quality (e.g., slow-twitch muscle conversion) is also expected to improve the balance ability of a subject. Improving muscle quality (e.g., slow-twitch muscle conversion) is also expected to improve the muscle strength of a subject. Specifically, for example, if the total muscle mass increases with an increase in the amount of slow-twitch muscle, it is expected to improve the muscle strength of a subject. Improving muscle quality (e.g., slow-twitch muscle conversion) is also expected to improve the walking speed of a subject. Specifically, for example, if the total muscle mass increases with an increase in the amount of slow-twitch muscle, it is expected to improve the walking speed of a subject. That is, effects based on improved muscle quality (e.g., slow-twitch muscle) include improved body composition, improved endurance, improved metabolism, improved balance, improved muscle strength, and improved walking speed. Improvements in body composition include increased muscle mass and decreased fat mass. The term "muscle mass" in body composition may refer to the absolute amount of muscle, the relative amount of muscle (i.e., the ratio of muscle weight to body weight), or both. The term "fat mass" in body composition may refer to the absolute amount of fat, the relative amount of fat (i.e., the ratio of fat weight to body weight), or both. Body composition (e.g., muscle mass and fat mass) can be measured using a body composition analyzer such as the EchoMRI-100 body composition analyzer (manufactured by Nippon Raytec Co., Ltd.). "Balance ability" may refer to the ability to maintain or recover posture.

[0020] In one embodiment, the effects of increasing the amount of mitochondria in muscles, improving body composition, improving endurance, improving metabolism, toning the body, and improving basic physical strength can all be effects based on improved energy metabolism. In one embodiment, the effects of improving balance ability, improving muscle strength, improving walking speed, improving training effects, and improving rehabilitation effects can all be effects based on improved muscle strength.

[0021] Improving muscle quality (e.g., slow-twitch muscle) is expected to improve body composition and / or metabolism, thereby, for example, toning a subject's body. Improving muscle quality (e.g., slow-twitch muscle) is expected to improve endurance, metabolism, and / or muscle strength, thereby improving the basic physical fitness of subjects, such as those who are physically inactive. Specifically, for example, when the amount of mitochondria in muscle increases with an increase in the amount of slow-twitch muscle, it is expected that the subject's endurance, metabolism, and / or muscle strength will improve, thereby improving the subject's basic physical fitness. Improving muscle quality (e.g., slow-twitch muscle) is expected to improve endurance, metabolism, and / or muscle strength, thereby improving the training effectiveness of subjects, such as exercise enthusiasts and athletes. Improving muscle quality (e.g., slow-twitch muscle) is expected to improve muscle strength and / or walking speed, thereby improving the rehabilitation effectiveness of subjects, such as those with mobility disorders, such as gait disorders. Furthermore, improving muscle quality (e.g., slow-twitch muscle formation), particularly through improved metabolism, is expected to enable, for example, regulating a subject's body temperature (e.g., maintaining or increasing body temperature). Improving muscle quality (e.g., slow-twitch muscle formation), particularly through improved metabolism, is expected to enable, for example, regulating a subject's blood glucose level (e.g., lowering blood glucose levels or suppressing increases in blood glucose levels). Improving muscle quality (e.g., slow-twitch muscle formation), particularly through improved metabolism, is expected to enable, for example, regulating a subject's blood lipid levels (e.g., lowering blood lipid levels or suppressing increases in blood lipid levels). That is, effects based on improving muscle quality (e.g., slow-twitch muscle formation) include toning the body, improving basic physical strength, improving training effects, improving rehabilitation effects, regulating a subject's body temperature, regulating a subject's blood glucose levels, and regulating a subject's blood lipid levels. "Training" may refer to physical training. "Rehabilitation" may refer to rehabilitation of physical functions.

[0022] It is expected that improving muscle quality (e.g., slow-twitching of muscles) will enable, for example, the prevention, improvement, and / or treatment of undesirable muscle-related symptoms in a subject. That is, effects based on improving muscle quality (e.g., slow-twitching of muscles) include the prevention, improvement, and / or treatment of undesirable muscle-related symptoms. Undesirable muscle-related symptoms may or may not be diseases. Undesirable muscle-related symptoms may develop or progress due to any of genetic factors, acquired factors, aging, etc.

[0023] Undesirable muscle-related symptoms include symptoms related to muscle atrophy and / or muscle weakness. "Muscle atrophy" may refer to a decrease in absolute muscle mass. Muscle atrophy includes atrophy of skeletal muscles. Specific examples of muscle atrophy include atrophy of slow-twitch and / or fast-twitch muscles. Muscle atrophy particularly includes atrophy of slow-twitch muscles. Muscle atrophy may result in, for example, a decrease in muscle strength. In other words, muscle weakness may be caused by, for example, muscle atrophy. Symptoms related to muscle atrophy and / or muscle weakness include symptoms caused by muscle atrophy and / or muscle weakness and symptoms accompanied by muscle atrophy and / or muscle weakness. Note that symptoms related to muscle atrophy and / or muscle weakness also include muscle atrophy itself and muscle weakness itself. Specific examples of undesirable muscle-related symptoms (e.g., symptoms related to muscle atrophy and / or muscle weakness) include disuse muscle atrophy, neurogenic muscle atrophy, and myogenic muscle atrophy. Disuse muscle atrophy includes sarcopenia (muscle atrophy due to aging) and muscle atrophy due to lack of muscle use. Muscle atrophy due to lack of use can be caused by being bedridden, inactivity, weightless flight, or immobilization. Immobilization can include limb immobilization for injury treatment. Neurogenic muscle atrophy includes amyotrophic lateral sclerosis (ALS), primary lateral sclerosis (PLS), spinal muscular atrophy (SMA), spinal and bulbar muscular atrophy (SBMA), poliomyelitis (polio), and Guillain-Barré syndrome (GBS). Myogenic muscle atrophy includes muscular dystrophy and polymyositis (PM).Specific examples of undesirable muscle-related symptoms (e.g., symptoms associated with muscle atrophy and / or muscle weakness) include myasthenia gravis (MG), cachexia, frailty, locomotive syndrome, and metabolic syndrome. Specific examples of undesirable muscle-related symptoms (e.g., symptoms associated with muscle atrophy and / or muscle weakness) include a decrease in basic physical fitness and a decrease in walking speed. A decrease in basic physical fitness or a decrease in walking speed may be due to, for example, aging or lack of muscle use.

[0024] Undesirable muscle-related symptoms also include posture-related symptoms, which include symptoms caused by poor posture. Undesirable muscle-related symptoms (e.g., posture-related symptoms) include lower back pain, stiff shoulders, and headaches.

[0025] Muscle-related undesirable symptoms also include symptoms related to mitochondrial function or metabolism. Mitochondrial function or metabolism-related symptoms include symptoms caused by insufficient or decreased mitochondrial function and symptoms caused by insufficient or decreased metabolism. Muscle-related undesirable symptoms (e.g., symptoms related to mitochondrial function or metabolism) include hyperglycemia, hyperlipidemia, vascular disorders, hypothermia, sensitivity to cold, and metabolic syndrome. Vascular disorders include arteriosclerosis. Prevention, amelioration, and / or treatment of vascular disorders may be, for example, prevention, amelioration, and / or treatment of hyperglycemia and / or hyperlipidemia. Prevention, amelioration, and / or treatment of vascular disorders may, for example, improve blood flow.

[0026] By improving muscle quality (e.g., slow-twitch muscle), for example, one of the effects based on improving muscle quality (e.g., slow-twitch muscle) as exemplified above may be obtained, or two or more of the effects based on improving muscle quality (e.g., slow-twitch muscle) as exemplified above may be obtained.

[0027] The active ingredient may or may not be used in combination with other techniques that can achieve the effects exemplified above, such as the muscle quality improving effect. Such techniques include physical exercise. That is, for example, the subject to which the active ingredient is administered may perform physical exercise. Physical exercise includes sports and training. Specific examples of physical exercise include aerobic exercise such as walking, jogging, yoga, and swimming, and anaerobic exercise (muscle training) such as weight training, squats, and push-ups. By using the active ingredient in combination with such other techniques, the effects of the active ingredient (e.g., the effects exemplified above, such as the muscle quality improving effect) may be further enhanced compared to when the active ingredient is used alone. Furthermore, by using the active ingredient in combination with such other techniques, the effects of such other techniques (e.g., the effects exemplified above, such as the muscle quality improving effect) may be further enhanced compared to when such other techniques are used alone. By using the active ingredient in combination with such other techniques, compared to when the active ingredient is used alone, for example, the effect of improving muscle strength, the effect of improving balance ability, the effect of improving training effect, the effect of improving rehabilitation effect, and / or the effect of improving walking speed may be further enhanced.In addition, by using the active ingredient in combination with such other techniques, compared to when the active ingredient is used alone, for example, muscle strength may be improved earlier.In addition, by using the active ingredient in combination with such other techniques, compared to when the active ingredient is used alone, for example, muscle strength may be maintained at a high level.

[0028] The active ingredient may be used for therapeutic or non-therapeutic purposes. That is, unless otherwise specified, the effects exemplified above, such as the muscle quality improving effect, may be obtained for therapeutic or non-therapeutic purposes.

[0029] "Therapeutic purposes" may refer to a concept including, for example, medical procedures, and specifically, may refer to a concept including treatment of the human body through therapy.

[0030] "Non-therapeutic purposes" may refer to, for example, a concept that does not include medical procedures, and specifically, a concept that does not include treatment of the human body through therapy. Non-therapeutic purposes include health promotion purposes.

[0031] "Prevention of symptoms" may mean, for example, preventing or delaying the onset of symptoms, or reducing the likelihood of onset of symptoms. "Amelioration of symptoms" or "treatment of symptoms" may mean, for example, improving symptoms, preventing or delaying worsening of symptoms, or preventing or delaying progression of symptoms. "Amelioration of symptoms" may particularly refer to these events collectively, but not for therapeutic purposes. "Treatment of symptoms" may particularly refer to these events collectively, but for therapeutic purposes.

[0032] "Cholinergic compounds" collectively refers to choline and its peripheral metabolites. Examples of cholineergic compounds include sn-glycero-3-phosphocholine (αGPC), citicoline, choline, choline phosphate, phosphocholine, phosphatidylcholine, sphingomyelin, and serine. Examples of cholineergic compounds in particular include αGPC, citicoline, and serine. Examples of cholineergic compounds in particular include αGPC. As the cholineergic compound, one type of component may be used, or two or more types of components may be used in combination.

[0033] Unless otherwise specified, serine may be in the D- or L-form, or a combination thereof. The ratio of the D- to L-form in the combination is not particularly limited. The ratio of the D- or L-form in the combination may be, for example, 20 to 80%, 30 to 70%, 40 to 60%, or 45 to 55% by molar ratio. Serine may particularly be in the L-form. Note that when D- or L-serine is selected, it is sufficient to use D- or L-serine as the active ingredient, and this does not preclude the use of L- or D-serine in combination.

[0034] When an active ingredient can form a salt, the active ingredient may be used in its free form, its salt, or a combination thereof. That is, unless otherwise specified, the term "active ingredient" may refer to the active ingredient in its free form, its salt, or a combination thereof. "Free form" means a form in which no salt is formed. Also, when an active ingredient can form a hydrate, the active ingredient may be used in its non-hydrate form, its hydrate, or a combination thereof. That is, the term "active ingredient" (e.g., "active ingredient in its free form" or "salt of the active ingredient") may include both the non-hydrate and the hydrate unless otherwise specified. The active ingredient may be in any form, such as an ion, when used.

[0035] The salt is not particularly limited as long as it is acceptable depending on the mode of use of the active ingredient.For example, when the active ingredient is orally ingested, the salt is not particularly limited as long as it is orally ingestible.For example, salts of acidic groups such as carboxyl groups include ammonium salts, salts with alkali metals such as sodium and potassium, salts with alkaline earth metals such as calcium and magnesium, aluminum salts, zinc salts, salts with organic amines such as triethylamine, ethanolamine, morpholine, pyrrolidine, piperidine, piperazine, and dicyclohexylamine, and salts with basic amino acids such as arginine and lysine. Specific examples of salts with basic groups such as amino groups include salts with inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, and hydrobromic acid, salts with organic carboxylic acids such as acetic acid, citric acid, benzoic acid, maleic acid, fumaric acid, tartaric acid, succinic acid, tannic acid, butyric acid, hybenzic acid, pamoic acid, enanthic acid, decanoic acid, teoclic acid, salicylic acid, lactic acid, oxalic acid, mandelic acid, malic acid, methylmalonic acid, and adipic acid, and salts with organic sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid. One type of salt may be used, or two or more types of salts may be used in combination.

[0036] The active ingredient may be a commercially available product or may be obtained by appropriate manufacturing. The method for manufacturing the active ingredient is not particularly limited. The active ingredient can be manufactured, for example, by chemical synthesis, enzymatic reaction, fermentation, extraction, or a combination thereof. The active ingredient may be purified to a desired degree or not. That is, the active ingredient may be a purified product, or a material containing the active ingredient. For example, the active ingredient may contain 1% (w / w) or more, 5% (w / w) or more, 10% (w / w) or more, 30% (w / w) or more, 50% (w / w) or more, 70% (w / w) or more, 90% (w / w) or more, or 95% (w / w) or more of the active ingredient.

[0037] When a material containing an active ingredient is used, the amount of the active ingredient (e.g., content (concentration) or amount used) is calculated based on the amount of the active ingredient itself in the material. When the active ingredient forms a salt or hydrate, the amount of the active ingredient (e.g., content (concentration) or amount used) is calculated based on the mass of the salt or hydrate converted to the mass of an equimolar amount of the free, non-hydrate form.

[0038] <2> Composition of the Present Invention The composition of the present invention is a composition containing an active ingredient (i.e., a cholinergic compound).

[0039] The composition of the present invention can be used, for example, by administering it to a subject. The composition of the present invention (e.g., the composition of the present invention administered to a subject) may be, for example, a pharmaceutical composition, a food composition, or a feed composition. The compositions of the present invention that are pharmaceutical compositions, food compositions, or feed compositions are also referred to as the "pharmaceutical composition of the present invention," the "food composition of the present invention," and the "feed composition of the present invention," respectively. Specifically, the composition of the present invention can be administered to a subject, for example, in the manner described in the method of the present invention described below. The description of the subject to which the active ingredient in the "method of the present invention" is administered can be applied mutatis mutandis to the subject to which the composition of the present invention is administered. The composition of the present invention may be administered to a subject, for example, to obtain the effects exemplified above, such as the muscle quality improvement effect.

[0040] As described above, the active ingredient may or may not be used in combination with other techniques that can achieve the effects exemplified above, such as the muscle quality improving effect. Thus, the composition of the present invention may be used in combination with such techniques. For example, the composition of the present invention may be used in combination with physical exercise. That is, for example, the subject to whom the composition of the present invention is administered may perform physical exercise.

[0041] "Pharmaceutical composition" may refer to a composition used for medical purposes, specifically a composition administered to a subject as a pharmaceutical. The term "pharmaceutical composition" may be used interchangeably or equivalently with the term "medicine." The pharmaceutical composition may be for oral administration or parenteral administration.

[0042] "Food composition" may mean a composition used for food applications, and specifically may mean a composition consumed by humans. A food composition may be, for example, a food or a food additive. "Food additive" may mean a composition added to food for use. A food additive may be, for example, added when the food is consumed. A food additive may be, for example, used in the production of food as a food ingredient. Food also includes beverages. Food also includes seasonings. Food is not limited to general foods, but also includes so-called health foods or medical foods such as nutritional supplements, nutritionally functional foods, and foods for specified health uses.

[0043] The term "feed composition" may refer to a composition used for feed purposes, specifically a composition to be consumed by organisms other than humans. The feed composition may be, for example, a feed or a feed additive. The term "feed additive" may refer to a composition to be added to feed. The feed additive may be, for example, added when feeding the feed. The feed additive may be, for example, used as a feed ingredient in the production of feed.

[0044] The composition of the present invention can be used, for example, for culturing animal cells. That is, the composition of the present invention may be, for example, a composition for animal cell culture. A composition of the present invention that is a composition for animal cell culture is also referred to as a "composition for animal cell culture of the present invention." Specifically, the composition of the present invention can be used, for example, for culturing animal cells in the manner described in the method of the present invention described below. The description of the animal cells in the "method of the present invention" can be applied mutatis mutandis to the animal cells cultured using the composition of the present invention. The composition of the present invention may be used, for example, for culturing animal cells in order to obtain the effects exemplified above, such as the muscle quality improvement effect.

[0045] The composition for animal cell culture may be, for example, a culture medium. Examples of the culture medium include a basal medium, a feed medium, and a perfusion medium. The term "basal medium" may refer to the medium used at the start of culture. The basal medium is also called an "initial medium." The term "fed-batch medium" may refer to the medium supplied to a culture system after the start of culture in fed-batch culture. The term "perfusion medium" may refer to the medium supplied to a culture system after the start of culture in continuous culture (not limited to perfusion culture).

[0046] The composition for animal cell culture may be, for example, a medium additive. The term "medium additive" may refer to a composition that is added to a medium before use. Examples of media to which the medium additive is added include basal medium, feed medium, and perfusion medium.

[0047] By utilizing the composition of the present invention, muscle quality can be improved (e.g., muscle can be made slow-twitch), i.e., a muscle quality improving effect (e.g., a slow-twitch effect) can be obtained. For example, by administering the composition of the present invention to a subject, a muscle quality improving effect (e.g., a slow-twitch effect) can be obtained in the subject. Furthermore, for example, by culturing animal cells using the composition of the present invention, a muscle quality improving effect (e.g., a slow-twitch effect) can be obtained in muscle tissue (e.g., cultured meat) formed by the cultured cells. In other words, the composition of the present invention (e.g., a food composition, a pharmaceutical composition, a feed composition, or a composition for animal cell culture) can be a composition for improving muscle quality (e.g., a composition for slow-twitch muscle).

[0048] Furthermore, by using the composition of the present invention, an effect based on improved muscle quality (e.g., slow-twitch muscle formation) may be obtained. For example, by administering the composition of the present invention to a subject, an effect based on improved muscle quality (e.g., slow-twitch muscle formation) may be obtained in the subject. That is, the composition of the present invention may be a composition for obtaining an effect based on improved muscle quality (e.g., slow-twitch muscle formation). A composition for obtaining an effect based on improved muscle quality (e.g., slow-twitch muscle formation) may be an example of a composition for improving muscle quality (e.g., a composition for slow-twitch muscle formation). Examples of compositions for obtaining an effect based on improved muscle quality (e.g., slow-twitch muscle formation) include compositions for increasing the amount of mitochondria in muscle, compositions for improving body composition, compositions for improving endurance, compositions for improving metabolism, compositions for improving balance ability, compositions for improving muscle strength, compositions for improving walking speed, compositions for toning the body, compositions for improving basic physical fitness, compositions for improving training effects, compositions for improving rehabilitation effects, compositions for regulating body temperature, compositions for regulating blood glucose levels, compositions for regulating blood lipid levels, and compositions for preventing, ameliorating, and / or treating undesirable muscle-related symptoms.

[0049] The composition of the present invention may consist of an active ingredient, or may contain ingredients other than the active ingredient. Ingredients other than the active ingredient are also referred to as "additional ingredients." As the additional ingredient, one type of ingredient may be used, or two or more types of ingredients may be used in combination.

[0050] The additional component is not particularly limited as long as it does not impair the object of the present invention. The additional component can be appropriately selected depending on various conditions, such as the type of the composition of the present invention and the mode of use. Examples of the additional component include components to be added to pharmaceuticals, foods, feeds, and culture media.

[0051] The composition of the present invention can be prepared, for example, by appropriately mixing the active ingredient and any additional ingredients.

[0052] The composition of the present invention may be formulated as appropriate. For formulation, additives (e.g., physiologically and / or pharmacologically acceptable additives) may be used as appropriate. Examples of additives include excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents, diluents, surfactants, and solvents. The additives can be selected as appropriate depending on various conditions, such as the type and mode of use of the composition of the present invention.

[0053] Examples of excipients include sugars or derivatives thereof such as lactose, sucrose, glucose, mannitol, and sorbitol; starches or derivatives thereof such as corn starch, potato starch, α-starch, dextrin, and carboxymethyl starch; celluloses or derivatives thereof such as crystalline cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, carboxymethyl cellulose, and calcium carboxymethyl cellulose; polysaccharides such as gum arabic, dextran, and pullulan; silicates such as light anhydrous silicic acid, synthetic aluminum silicate, and magnesium aluminometasilicate; phosphates such as calcium phosphate; carbonates such as calcium carbonate; and sulfates such as calcium sulfate.

[0054] Examples of binders include gelatin, polyvinylpyrrolidone, and macrogol in addition to the above-mentioned excipients.

[0055] Disintegrants include, in addition to the above-mentioned excipients, croscarmellose sodium, carboxymethyl starch sodium, and cross-linked polyvinylpyrrolidone.

[0056] Examples of lubricants include talc; stearic acid; stearates such as calcium stearate and magnesium stearate; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; and waxes such as carnauba wax.

[0057] Examples of stabilizers include parahydroxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; and sorbic acid.

[0058] Flavoring agents include sweeteners, acidulants, and fragrances.

[0059] The form of the composition of the present invention is not particularly limited. The form of the composition of the present invention can be appropriately selected depending on various conditions, such as the type of composition of the present invention and the mode of use. The composition of the present invention may be in any form, such as powder, granules, flakes, tablets, capsules, paste, or liquid. For example, the dosage form of the pharmaceutical composition for oral administration includes solid formulations such as powders, granules, tablets, and capsules, and liquid formulations such as solutions, syrups, suspensions, and emulsions. Furthermore, for parenteral administration, the dosage form of the pharmaceutical composition includes solid formulations such as suppositories and ointments, and liquid formulations such as injections.

[0060] The content and content ratio of each component (i.e., active component and optional additional component) in the composition of the present invention are not particularly limited as long as the desired effect, such as muscle quality improvement effect, is obtained. The content and content ratio of each component in the composition of the present invention can be appropriately set depending on various conditions, such as the type of composition of the present invention and the mode of use.

[0061] The content of the active ingredient in the composition of the present invention is more than 0% (w / w) and not more than 100% (w / w). The content of the active ingredient in the composition of the present invention may be, for example, 100 ppm (w / w) or more, 200 ppm (w / w) or more, 500 ppm (w / w) or more, 1000 ppm (w / w) or more, 2000 ppm (w / w) or more, 5000 ppm (w / w) or more, 1% (w / w) or more, 2% (w / w) or more, 5% (w / w) or more, 10% (w / w) or more, 20% (w / w) or more, 30% (w / w) or more, 50% (w / w) or more, or 70% (w / w) or more, and may be 100% (w / w) or less, less than 100% (w / w), 99.9% (w / w) or less, 90% (w / w) or less, 70% (w / w) or less, 50% (w / w) or less, 30% (w / w) or less, or 70% (w / w) or more. % (w / w) or less, 20% (w / w) or less, 10% (w / w) or less, 5% (w / w) or less, 2% (w / w) or less, 1% (w / w) or less, 5000 ppm (w / w) or less, 2000 ppm (w / w) or less, 1000 ppm (w / w) or less, or 500 ppm (w / w) or less, or any compatible combination thereof. The content of the active ingredient in the composition of the present invention may be, specifically, for example, 100 ppm (w / w) to 1000 ppm (w / w), 1000 ppm (w / w) to 1% (w / w), 1% (w / w) to 10% (w / w), 10% (w / w) to 30% (w / w), 30% (w / w) to 50% (w / w), 50% (w / w) to 70% (w / w), or 70% (w / w) to 99.9% (w / w).Specific examples of the content of the active ingredient in the composition of the present invention include 100 ppm (w / w) to 99.9% (w / w), 200 ppm (w / w) to 99.9% (w / w), 500 ppm (w / w) to 99.9% (w / w), 1000 ppm (w / w) to 99.9% (w / w), 1% (w / w) to 99.9% (w / w), 100 ppm (w / w) to 50% (w / w), 100 ppm (w / w) to 10% (w / w), 100 ppm (w / w) to 1% (w / w), 1000 ppm (w / w) to 50% (w / w), 1000 ppm (w / w) to 10% (w / w), 1000 ppm (w / w) to 1% (w / w), 1 % (w / w) to 50% (w / w), or 1% (w / w) to 10% (w / w). When the composition of the present invention contains two or more cholinergic compounds (i.e., two or more active ingredients), the contents of those two or more cholinergic compounds in the composition of the present invention may be set independently or in total within the range of the content of the active ingredients in the composition of the present invention exemplified above (provided that the total content of those two or more cholinergic compounds in the composition of the present invention is 100% (w / w) or less). When the composition of the present invention contains two or more cholinergic compounds, the "content of the active ingredients in the composition of the present invention" means the total content of those two or more cholinergic compounds in the composition of the present invention, unless otherwise specified.

[0062] Furthermore, the content of each component (i.e., the active component and any additional component) in the composition of the present invention can be set, for example, so as to obtain the amount of each component to be administered to a subject in the method of the present invention described below.

[0063] Furthermore, the content of each component (i.e., the active component and any additional component) in the composition of the present invention can be set, for example, so as to obtain the concentration of each component in the medium in the method of the present invention described below.

[0064] The components (i.e., the active ingredient and optional additional ingredients) contained in the composition of the present invention may be mixed together and contained in the composition of the present invention, or may be contained separately or in any combination. For example, the composition of the present invention may be provided as a set of components each packaged separately. In such a case, the components contained in the set can be used together as appropriate when used.

[0065] The composition of the present invention may be provided as a product labeled with the above-mentioned uses, such as improving muscle quality, slow-twitch muscle, increasing the amount of mitochondria in muscle, improving body composition, improving endurance, improving metabolism, improving balance, improving muscle strength, improving walking speed, toning the body, improving basic physical fitness, improving training effects, improving rehabilitation effects, regulating body temperature, regulating blood glucose levels, regulating blood lipid levels, and preventing, improving, and / or treating undesirable muscle-related symptoms. "Indication of use" includes all actions for informing consumers of the use. "Indication of use" is not limited to direct indication of the use, but also includes displaying a phrase that can remind or lead consumers to infer the use. Examples of phrases that can remind or lead consumers to infer the above-mentioned uses include phrases related to the intended recipient, such as "athletes," "sports enthusiasts," "people who are not getting enough exercise," "people concerned about muscle weakness," and "people concerned about endurance."

[0066] <3> Method of the Present Invention The method of the present invention is a method comprising a step of utilizing an active ingredient (i.e., a cholinergic compound).

[0067] The active ingredient can be used, for example, by administering it to a subject. That is, the method of the present invention may be, for example, a method comprising a step of administering the active ingredient (i.e., a cholinergic compound) to a subject. This step is also referred to as an "administration step."

[0068] The active ingredient can be used, for example, in culturing animal cells. That is, the method of the present invention may be, for example, a method comprising a step of culturing animal cells in the presence of the active ingredient (i.e., a cholinergic compound). This step is also referred to as a "culturing step." Furthermore, the method of the present invention may be, for example, a method of culturing animal cells.

[0069] The method of the present invention, specifically by utilizing the active ingredient, can improve muscle quality (e.g., convert muscle to slow-twitch muscle), i.e., a muscle quality improving effect (e.g., a slow-twitch muscle conversion effect) can be obtained. For example, the method of the present invention, specifically by administering the active ingredient to a subject, can achieve a muscle quality improving effect (e.g., a slow-twitch muscle conversion effect) in the subject. Furthermore, for example, by culturing animal cells using the active ingredient, a muscle quality improving effect (e.g., a slow-twitch muscle conversion effect) can be achieved in muscle tissue (e.g., cultured meat) formed by the cultured cells. In other words, the method of the present invention can be a method of improving muscle quality (e.g., a method of converting muscle to slow-twitch muscle).

[0070] Furthermore, the method of the present invention, specifically by utilizing the active ingredient, may provide an effect based on improved muscle quality (e.g., slow-twitch muscle). For example, the method of the present invention, specifically by administering the active ingredient to a subject, may provide an effect based on improved muscle quality (e.g., slow-twitch muscle). That is, the method of the present invention may be a method for obtaining an effect based on improved muscle quality (e.g., slow-twitch muscle). A method for obtaining an effect based on improved muscle quality (e.g., slow-twitch muscle) may be an example of a method for improving muscle quality (e.g., a method for slow-twitch muscle). Examples of methods for obtaining an effect based on improved muscle quality (e.g., slow-twitch muscle) include methods for increasing the amount of mitochondria in muscle, improving body composition, improving endurance, improving metabolism, improving balance ability, improving muscle strength, improving walking speed, toning the body, improving basic physical fitness, improving training effects, improving rehabilitation effects, regulating body temperature, regulating blood glucose levels, regulating blood lipid concentrations, and methods for preventing, ameliorating, and / or treating undesirable muscle-related symptoms.

[0071] Muscle tissue may also be produced by culturing animal cells. That is, muscle tissue may be produced by forming muscle tissue from cultured cells. Examples of muscle tissue include cultured meat. That is, the method of the present invention (e.g., a method of culturing animal cells) may be a method for producing muscle tissue (e.g., cultured meat). Cultured meat may consist of muscle tissue, or may contain other tissues. Examples of other tissues include adipose tissue. The muscle tissue (e.g., cultured meat) produced may be muscle tissue (e.g., cultured meat) with improved muscle quality.

[0072] The term "administering an active ingredient to a subject" can be used interchangeably or equivalently with the term "allowing a subject to ingest an active ingredient." Intake of the active ingredient can be voluntary (i.e., free intake) or forced (i.e., gavage). That is, the administration step can be, for example, a step of providing the active ingredient (which can be incorporated into food or feed) to the subject, thereby allowing the subject to ingest the active ingredient ad libitum. Administration can be oral or parenteral. Administration can particularly be oral. Parenteral administration includes injection, tube administration, rectal administration, and intranasal administration.

[0073] The mode of administration of the active ingredient (e.g., the subject, the timing of administration, the duration of administration, the number of administrations, the dosage, and other conditions related to administration) is not particularly limited as long as the desired effect, such as the muscle quality improvement effect, is obtained. The mode of administration of the active ingredient can be appropriately determined depending on various conditions, such as the type, age, and health condition of the subject.

[0074] The subject to which the active ingredient is administered is not particularly limited as long as the desired effects, such as muscle quality improvement, are obtained. Examples of subjects to which the active ingredient is administered include mammals. Examples of mammals include primates such as humans, monkeys, and chimpanzees, rodents such as mice, rats, hamsters, and guinea pigs, and various other mammals such as rabbits, horses, cows, sheep, goats, pigs, dogs, and cats. Mammals include humans in particular. The conditions of the subject to which the active ingredient is administered (e.g., gender, age, health status, and other conditions related to the subject) are not particularly limited as long as the desired effects, such as muscle quality improvement, are obtained. The subject to which the active ingredient is administered may be either a male or a female subject. The subject to which the active ingredient is administered may be of any age, such as an infant, child, adult, middle-aged or elderly person, or elderly person. The subject to which the active ingredient is administered may be, for example, a healthy subject or an unhealthy subject. An unhealthy subject may be a subject exhibiting undesirable muscle-related symptoms. In the case of humans, the subjects to which the active ingredient is administered may or may not be, for example, athletes, sports enthusiasts, or other people who perform physical exercise other than daily activities.

[0075] The dosage of the active ingredient may be, for example, 0.01 mg / kg body weight / day or more, 0.02 mg / kg body weight / day or more, 0.05 mg / kg body weight / day or more, 0.1 mg / kg body weight / day or more, 0.2 mg / kg body weight / day or more, 0.5 mg / kg body weight / day or more, 1 mg / kg body weight / day or more, 2 mg / kg body weight / day or more, 5 mg / kg body weight / day or more, 10 mg / kg body weight / day or more, 20 mg / kg body weight / day or more, 50 mg / kg body weight / day or more, 100 mg / kg body weight / day or more, 200 mg / kg body weight / day or more, or 500 mg / kg body weight / day or more, and may be 1000 mg / kg body weight / day or less, 500 mg / kg body weight / day or less, 200 mg / kg body weight / day or less, 100 mg / kg body weight / day or less, 50 mg / kg body weight / day or less, 2 ... The dose may be less than or equal to 5 mg / kg body weight / day, less than or equal to 2 mg / kg body weight / day, less than or equal to 1 mg / kg body weight / day, less than or equal to 0.5 mg / kg body weight / day, less than or equal to 0.2 mg / kg body weight / day, less than or equal to 0.1 mg / kg body weight / day, less than or equal to 0.05 mg / kg body weight / day, or less than or equal to 0.02 mg / kg body weight / day, or any compatible combination thereof. The dosage of the active ingredient may be, for example, 0.01 to 0.02 mg / kg body weight / day, 0.02 to 0.05 mg / kg body weight / day, 0.05 to 0.1 mg / kg body weight / day, 0.1 to 0.2 mg / kg body weight / day, 0.2 to 0.5 mg / kg body weight / day, 0.5 to 1 mg / kg body weight / day, 1 to 2 mg / kg body weight / day, 2 to 5 mg / kg body weight / day, 5 to 10 mg / kg body weight / day, 10 to 20 mg / kg body weight / day, 20 to 50 mg / kg body weight / day, 50 to 100 mg / kg body weight / day, 100 to 200 mg / kg body weight / day, 200 to 500 mg / kg body weight / day, or 500 to 1000 mg / kg body weight / day. The dosage of the active ingredient may be specifically, for example, 0.01 to 1000 mg / kg body weight / day, 0.05 to 200 mg / kg body weight / day, or 0.2 to 50 mg / kg body weight / day.When two or more cholinergic compounds (i.e., two or more active ingredients) are administered, the dosages of the two or more cholinergic compounds may be set independently or in total within the dosage range of the active ingredient exemplified above. When two or more cholinergic compounds are administered, the "dosage of active ingredients" refers to the total dosage of the two or more cholinergic compounds, unless otherwise specified.

[0076] The administration period of the active ingredient may be, for example, 1 day or more, 3 days or more, 1 week or more, 2 weeks or more, 4 weeks or more, 2 months or more, 3 months or more, 4 months or more, 6 months or more, 9 months or more, or 12 months or more, or 10 years or less, 5 years or less, 1 year or less, or 6 months or less, or any compatible combination thereof. The active ingredient may be administered, for example, throughout the subject's lifetime or for a portion of the subject's lifetime. The active ingredient may be administered, for example, at least until a desired effect, such as a muscle quality improvement effect, is achieved. The active ingredient may be administered, for example, daily or once every few days. The active ingredient may particularly be administered daily. The dosage of the active ingredient at each administration may be constant or not.

[0077] For example, the active ingredient may be administered to a subject directly, or may be prepared as a composition containing the active ingredient, such as a pharmaceutical composition, food composition, or feed composition, and then administered to a subject. The description of the composition of the present invention applies mutatis mutandis to compositions containing the active ingredient. The active ingredient may be administered alone or in combination with additional ingredients. Examples of additional ingredients include pharmaceuticals, foods, feeds, and ingredients contained therein. Furthermore, the description of the additional ingredients contained in the composition of the present invention applies mutatis mutandis to additional ingredients used in the methods of the present invention.

[0078] The active ingredient can also be administered to a subject, for example, using the composition of the present invention (specifically, by administering the composition of the present invention to the subject). That is, one embodiment of the method of the present invention may be a method comprising administering the composition of the present invention to a subject. That is, "administration of an active ingredient" also encompasses administration of the composition of the present invention. The administration mode of the composition of the present invention (e.g., the subject, administration time, administration period, number of administrations, dosage, and other administration conditions) is not particularly limited as long as the desired effect, such as muscle quality improvement, is obtained. The administration mode of the composition of the present invention can be appropriately determined depending on various conditions, such as the type, age, and health condition of the subject. The description of the administration mode of the active ingredient can be applied mutatis mutandis to the administration mode of the composition of the present invention. That is, the composition of the present invention may be administered to, for example, subjects such as those exemplified above. Furthermore, the dosage of the composition of the present invention may be determined, for example, so as to obtain the dosage of the active ingredient as exemplified above. Furthermore, the composition of the present invention may be administered alone or in combination with additional components.

[0079] As described above, the active ingredient may or may not be used in combination with other techniques that can achieve the effects exemplified above, such as the muscle quality improving effect. Therefore, the method of the present invention may be used in combination with such techniques. In other words, such techniques may be implemented in the method of the present invention. The method of the present invention may be used in combination with, for example, physical exercise. In other words, in the method of the present invention, for example, physical exercise may be implemented, that is, the subject to whom the active ingredient is administered may perform physical exercise.

[0080] The animal cells are not particularly limited as long as they can form muscle tissue through culture. "Animal cells capable of forming muscle tissue through culture" may mean that the animal cells can differentiate through culture to form muscle tissue, specifically, that the animal cells can differentiate into myotube cells through culture to form muscle tissue. Examples of animal cells include stem cells such as mesenchymal stem cells, embryonic stem cells (ES cells), and induced pluripotent stem cells (iPS cells), as well as myoblasts. Examples of animals include mammals. Examples of mammals include primates such as humans, monkeys, and chimpanzees, rodents such as mice, rats, hamsters, and guinea pigs, and various other mammals such as rabbits, horses, cows, sheep, goats, pigs, dogs, and cats. When producing cultured meat for consumption through culture, mammals particularly include animals used for meat production, such as cows and pigs.

[0081] The animal cells are cultured in the presence of an active ingredient. "The animal cells are cultured in the presence of an active ingredient" may mean, for example, that the medium used for culturing the animal cells contains the active ingredient. "The animal cells are cultured in the presence of an active ingredient" may mean, for example, that the active ingredient is supplied to the culture system during the culturing of the animal cells.

[0082] Animal cell culture may be carried out using, for example, the composition of the present invention. By culturing animal cells using the composition of the present invention, the animal cell culture can be carried out in the presence of an active ingredient. The composition of the present invention may be used, for example, as a culture medium for culturing animal cells. That is, for example, when the composition of the present invention is a culture medium, animal cells may be cultured in the composition of the present invention (i.e., the culture medium). That is, "culturing animal cells using the composition of the present invention" may mean, for example, culturing animal cells in the composition of the present invention, which is a culture medium. Furthermore, the composition of the present invention may be used, for example, as a culture medium additive for culturing animal cells. That is, for example, when the composition of the present invention is a culture medium additive, the composition of the present invention (i.e., the culture medium additive) may be added to a culture medium, and animal cells may be cultured in the culture medium to which the composition of the present invention has been added. That is, "culturing animal cells using the composition of the present invention" may mean, for example, adding the composition of the present invention, which is a culture medium additive, to a culture medium, and culturing animal cells in the culture medium to which the composition of the present invention has been added.

[0083] The medium composition and culture conditions are not particularly limited as long as the culture is performed in the presence of an active ingredient and muscle tissue is formed by the cultured cells. The medium composition and culture conditions can be appropriately set depending on various conditions, such as the type of animal cells. Except for the presence of an active ingredient, the culture can be performed using, for example, a conventional medium and conventional conditions used for the formation of muscle tissue by differentiation of animal cells (e.g., the production of cultured meat), either as is or with appropriate modifications.

[0084] Culture can be performed by batch culture, fed-batch culture, continuous culture, or a combination thereof. Continuous cultures include perfusion culture and chemostat culture. The medium used at the start of culture is also called the "initial medium" or "basal medium." The medium supplied to a culture system (e.g., the initial medium) in fed-batch culture is also called the "feed medium." The medium supplied to a culture system (e.g., the initial medium) in continuous culture (not limited to perfusion culture) is also called the "perfusion medium." Supplying a feed medium or perfusion medium to a culture system in fed-batch or continuous culture is also simply called "medium supply." Medium supply may be performed throughout the entire culture period or only for a portion of the culture period. Medium supply may be performed continuously or intermittently. During culture (particularly continuous culture such as perfusion culture), the culture medium may be withdrawn. The culture medium may be withdrawn throughout the entire culture period, or only during a portion of the culture period. The culture medium may be withdrawn continuously or intermittently. The culture medium may be withdrawn and the medium may be supplied simultaneously, or not simultaneously. Culturing may be carried out three-dimensionally, for example, using a scaffold that matches the shape of the muscle tissue to be formed (e.g., cultivated meat).

[0085] The culture can be carried out, for example, using a liquid medium.

[0086] The media used for culture, for example, the basal medium, the feed medium, and the perfusion medium, can be selected independently.

[0087] The medium used for the culture may be a commercially available medium or an appropriately prepared medium.

[0088] The medium used for the culture includes a medium containing components essential for culturing animal cells (for example, a carbon source, a nitrogen source, inorganic salts, etc.).

[0089] Specific examples of media used for culture include Dulbecco's Modified Eagle's Medium (DMEM), Ham's Nutrient Mixture F12, DMEM / F12, McCoy's 5A medium, Minimum Essential Medium (MEM), Eagle's Minimum Essential Medium (EMEM), alpha Modified Eagle's Minimum Essential Medium (αMEM), Roswell Park Memorial Institute (RPMI) 1640 medium, Iscove's Modified Dulbecco's Medium (IMDM), MCDB131 medium, William's Medium E, and Fischer's Medium.

[0090] Furthermore, specific examples of media used for culture (for example, media particularly used for culturing stem cells (particularly pluripotent stem cells)) include STEMPRO (registered trademark) hESC SFM medium (Life Technologies), mTeSR1 medium (STEMCELL Technologies), TeSR2 medium (STEMCELL Technologies), TeSR-E8 medium (STEMCELL Technologies), Essential 8 medium (Life Technologies), HEScGRO (trademark) Serum-Free Medium for hES cells (Millipore), PluriSTEM (trademark) Human ES / iPS Medium (EMD Millipore), NutriStem (registered trademark) hESC XF medium (Biological Industries Israel Beit-Haemek), NutriStem (trademark) XF / FF Culture Medium (Stemgent), AF NutriStem (registered trademark) hESC XF medium (Biological Industries Israel Examples of suitable medium include hESF9 medium, hESF-FX medium, CDM medium, DEF-CS 500 Xeno-Free 3D Spheroid Culture Medium (Cellartis), and StemFlex medium (Thermo Fisher Scientific).

[0091] Other commercially available media include animal cell culture media such as CELLiST Basal Media BASAL3, BASAL4P, and BASAL10 (Ajinomoto Co., Inc.), Opti-MEM (Thermo Fisher Scientific), RPMI 1640 (Thermo Fisher Scientific), CD293 (Thermo Fisher Scientific), CHO-S-SFMII (Thermo Fisher Scientific), CHO-SF (Sigma-Aldrich), EX-CELL CD CHO (Sigma-Aldrich), EX-CELL™302 (Sigma-Aldrich), IS CHO-CD (Irvine Scientific), and IS CHO-CDXP (Irvine Scientific).

[0092] The media exemplified above may be used for culture by adding, for example, an active ingredient.

[0093] The medium used for culture may be, for example, the composition of the present invention (specifically, the composition of the present invention that is a medium). That is, when the composition of the present invention is a medium, the composition of the present invention may be used for culture as is, or may be appropriately prepared as a liquid medium of a desired composition. For example, the composition of the present invention may be diluted with an aqueous medium such as water or an aqueous buffer solution to prepare a liquid medium and use it for culture. The composition of the present invention may be used as one or more media selected from a basal medium, a feed medium, and a perfusion medium. That is, for example, in the case of fed-batch culture, the composition of the present invention may be used as one or both of the basal medium and the feed medium. Furthermore, for example, in the case of continuous culture, the composition of the present invention may be used as one or both of the basal medium and the perfusion medium.

[0094] Furthermore, the medium used for culture may be, for example, a medium supplemented with the composition of the present invention (specifically, the composition of the present invention as a medium additive). The medium to which the composition of the present invention is added may be a commercially available medium or an appropriately prepared medium. The composition of the present invention may be added to one or more media selected from a basal medium, a feed medium, and a perfusion medium. That is, for example, in the case of fed-batch culture, the composition of the present invention may be added to one or both of the basal medium and the feed medium. Furthermore, for example, in the case of continuous culture, the composition of the present invention may be added to one or both of the basal medium and the perfusion medium.

[0095] The medium may contain various medium components. Examples of medium components include carbon sources, amino acid sources, peptides, proteins, vitamins, fatty acids, lipids, inorganic components, pH buffers, growth factors, cytokines, hormones, cell adhesion factors, extracellular matrix components, serum, antibiotics, and gene expression inducers. Any of these medium components may be essential or effective for, for example, the survival or proliferation of animal cells. Any of these medium components may be pre-contained in the medium exemplified above, or may be added to the medium exemplified above.

[0096] Examples of carbon sources include sugars such as glucose, fructose, sucrose, and maltose.

[0097] The amino acid source may include amino acids. In addition, both peptides and proteins may be examples of amino acid sources. The amino acids may include glycine, alanine, valine, leucine, isoleucine, cysteine, methionine, phenylalanine, tyrosine, tryptophan, histidine, lysine, arginine, serine, threonine, aspartic acid, glutamic acid, asparagine, glutamine, proline, and ornithine. The amino acids may be, for example, L-form.

[0098] Examples of peptides include dipeptides and tripeptides. Specific examples of peptides include glycylglycylglycine and soybean peptide. The descriptions of amino acids also apply mutatis mutandis to the amino acids that make up peptides.

[0099] Examples of proteins include albumin and transferrin.

[0100] Vitamins include vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B12, vitamin C, vitamin D, vitamin E, and vitamin K, and precursors thereof.

[0101] Fatty acids include oleic acid, arachidonic acid, and linoleic acid.

[0102] Lipids include cholesterol.

[0103] Examples of inorganic components include sodium, potassium, calcium, magnesium, phosphorus, and various trace elements (e.g., Co, Cu, F, Fe, Mn, Mo, Ni, Se, Si, Ni, Bi, V, and Zn).Specific examples of inorganic components include inorganic salts such as sodium chloride, potassium chloride, calcium chloride, magnesium sulfate, and sodium dihydrogen phosphate.

[0104] Examples of pH buffers include sodium bicarbonate, phosphate, N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), and N-[tris(hydroxymethyl)methyl]glycine (Tricine).

[0105] Examples of growth factors include fibroblast growth factor (FGF), hepatocyte growth factor (HGF), epidermal growth factor (EGF), transforming growth factor (TGF)-α, transforming growth factor (TGF)-β, vascular endothelial growth factor (VEGF), activin A, and insulin-like growth factor-1 (IGF-1). Examples of FGF include basic fibroblast growth factor (bFGF). Examples of growth factors include bFGF in particular. That is, at least bFGF may be used as a medium component. That is, the medium may contain at least bFGF. Examples of bFGF include bovine basic fibroblast growth factor (bbFGF). "Bovine basic fibroblast growth factor (bbFGF)" may refer to bovine-derived bFGF. The term "bovine" in reference to bbFGF may refer to organisms of the genus Bos. Bos organisms include Bos taurus. bbFGF is not limited to bFGF found in organisms of the genus Bos, and may be a modified form thereof.

[0106] Cytokines include interleukins.

[0107] Hormones include dexamethasone, hydrocortisone, estradiol, progesterone, glucagon, and insulin.

[0108] Examples of cell adhesion factors or extracellular matrix components include Type I collagen, Type II collagen, fibronectin, laminin, poly-L-lysine, and poly-D-lysine.

[0109] Antibiotics include amphotericin B, kanamycin, gentamicin, streptomycin, and penicillin.

[0110] Various components, such as active ingredients, may be contained in the initial medium, feed medium, perfusion medium, or a combination thereof. That is, various components, such as active ingredients, may be supplied to the medium alone or in any combination during the culture process. These components may be supplied once, multiple times, or continuously. The compositions (e.g., the types and / or concentrations of the components) of the initial medium, feed medium, and perfusion medium may or may not be the same. That is, the types of components contained in the initial medium may or may not be the same as the types of components contained in the feed medium or perfusion medium. Furthermore, the concentrations of the components contained in the initial medium may or may not be the same as the concentrations of the components contained in the feed medium or perfusion medium. For example, when a feed medium is used for perfusion culture, the compositions of the initial medium and the feed medium may be the same. Furthermore, two or more feed media or perfusion media with different compositions (e.g., the types and / or concentrations of the components contained) may be used. For example, when the feed medium or perfusion medium is supplied intermittently multiple times, the composition of the feed medium or perfusion medium may or may not be the same for each supply. Furthermore, various components such as active ingredients may be supplied to the medium in a form not contained in the feed medium or perfusion medium, such as a powder. For example, various components such as active ingredients may be supplied to the medium by adding the composition of the present invention.

[0111] The seeding amount of animal cells at the start of culture is, for example, 1 × 10 in terms of viable cell number. 3 cells / mL or more, 1×10 4 cells / mL or more, 1×10 5 cells / mL or more, 1×10 6 cells / mL or more, or 1×10 7 cells / mL or more, but not more than 1 x 10 8 cells / mL or less, 1×10 7 cells / mL or less, 1×106 cells / mL or less, 1×10 5 cells / mL or less, or 1 x 10 4 The seeding amount of animal cells at the start of culture may be, for example, 1 × 10 cells / mL or less in terms of the number of viable cells. 3 ~1×10 4 cells / mL, 1 × 10 4 ~1×10 5 cells / mL, 1 × 10 5 ~1×10 6 cells / mL, 1 × 10 6 ~1×10 7 cells / mL, or 1 x 10 7 ~1×10 8 The seeding amount of animal cells at the start of culture may be, for example, 1 × 10 cells / mL in terms of the number of viable cells. 3 ~1×10 8 cells / mL, 1 × 10 4 ~1×10 7 cells / mL, or 1 x 10 5 ~1×10 6 The number of viable cells may be measured using, for example, a viable cell autoanalyzer, Vi-CELL TM Measurement can be performed using XR (Beckman Coulter).

[0112] The culture may be carried out under, for example, 5 to 15% CO ２ CO etc. ２The cultivation may be carried out under a pH-containing atmosphere. The pH of the medium may be, for example, near neutral. "Near neutral" may mean, for example, pH 6 to 8, pH 6.5 to 7.5, or pH 6.8 to 7.2. The pH of the medium can be adjusted as needed during cultivation. The pH of the medium can be adjusted using various alkaline or acidic substances such as ammonia gas, ammonia water, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, magnesium carbonate, sodium hydroxide, potassium hydroxide, calcium hydroxide, and magnesium hydroxide. The cultivation temperature may be, for example, 30 to 38°C. The culture period may be, for example, 0.5 days or more, 1 day or more, 2 days or more, 3 days or more, 4 days or more, 5 days or more, 6 days or more, 7 days or more, 8 days or more, 9 days or more, 10 days or more, 12 days or more, 15 days or more, or 20 days or more, or 60 days or less, 50 days or less, 40 days or less, 30 days or less, 25 days or less, 20 days or less, 15 days or less, 12 days or less, 10 days or less, 9 days or less, 8 days or less, or 7 days or less, or any compatible combination thereof. Specific examples of the culture period may be, for example, 1 to 60 days, 3 to 25 days, or 5 to 20 days. Culture may be continued, for example, until muscle tissue (e.g., cultured meat) has been formed to a desired extent.

[0113] The concentration of the active ingredient in the medium may be, for example, 0.01 mM or more, 0.02 mM or more, 0.05 mM or more, 0.1 mM or more, 0.2 mM or more, 0.5 mM or more, 1 mM or more, 2 mM or more, 5 mM or more, 10 mM or more, 15 mM or more, 20 mM or more, or 25 mM or more, or 30 mM or less, 25 mM or less, 20 mM or less, 15 mM or less, 10 mM or less, 5 mM or less, 2 mM or less, 1 mM or less, 0.5 mM or less, 0.2 mM or less, 0.1 mM or less, 0.05 mM or less, or 0.02 mM or less, or a compatible combination thereof. The concentration of the active ingredient in the medium may be, for example, 0.01 to 0.02 mM, 0.02 to 0.05 mM, 0.05 to 0.1 mM, 0.1 to 0.2 mM, 0.2 to 0.5 mM, 0.5 to 1 mM, 1 to 2 mM, 2 to 5 mM, 5 to 10 mM, 10 to 15 mM, 15 to 20 mM, 20 to 25 mM, or 25 to 30 mM. The concentration of the active ingredient in the medium may be, for example, 0.01 to 30 mM, 0.02 to 15 mM, or 0.05 to 5 mM. When the medium contains two or more cholinergic compounds (i.e., two or more active ingredients), the concentrations of the two or more cholinergic compounds in the medium may be set independently or in total within the range of the active ingredient concentration in the medium exemplified above. In addition, when the medium contains two or more cholinergic compounds, the "concentration of the active ingredient in the medium" means the total concentration of those two or more cholinergic compounds in the medium, unless otherwise specified.

[0114] Each of the various components, such as the active ingredient, may be contained in the medium throughout the entire culture period, or only during a portion of the culture period. In other words, "culture is carried out in a medium containing a certain component" or "a certain component is contained in the medium during culture" means that the component is contained in the medium during at least a portion of the culture period, and does not necessarily have to be contained in the medium throughout the entire culture period. Each of the various components, such as the active ingredient, may be contained in the medium at the start of culture, or may be supplied to the medium after the start of culture. Furthermore, each of the various components, such as the active ingredient, may be contained in the medium at the start of culture, and further supplied to the medium after the start of culture (e.g., after the active ingredient has been consumed).

[0115] Each of the various components, such as the active ingredient, may be contained in the medium at the concentrations exemplified above throughout the entire culture period, or only during a portion of the culture period. That is, "culture is carried out in a medium containing a certain component at a certain concentration," "a certain component is contained in the medium at a certain concentration during culture," or "the concentration of a certain component in the medium during culture is a certain concentration" means that the concentration of the component in the medium is within the range for at least a portion of the culture period, but does not necessarily need to be within the range for the entire culture period. Each of the various components, such as the active ingredient, may be contained in the medium at the concentrations exemplified above, for example, at the start of culture, or may be supplied to the medium after the start of culture to achieve the concentrations exemplified above. Furthermore, each of the various components, such as the active ingredient, may be contained in the medium at the concentrations exemplified above at the start of culture, and then further supplied to the medium after the start of culture (e.g., after the component has been consumed) to achieve the concentrations exemplified above.

[0116] The length of the "partial culture period" is not particularly limited as long as the desired effect, such as a muscle quality improvement effect, is obtained. The length of the "partial culture period" can be appropriately set depending on various conditions, such as the type of animal cells and the length of the culture period. The "partial culture period" may be, for example, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 95% or more, 97% or more, or 99% or more of the total culture period. Furthermore, the "partial culture period" may be, for example, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 95% or more, 97% or more, or 99% or more of the total culture period after the animal cells have differentiated into myotubes. Furthermore, a "partial period" may be, for example, a period of 0.5 days or more, 1 day or more, 2 days or more, 3 days or more, 4 days or more, 5 days or more, 6 days or more, 7 days or more, 8 days or more, 9 days or more, 10 days or more, 12 days or more, or 15 days or more.

[0117] Furthermore, the concentrations of various components, such as active ingredients, in the culture medium may be set to the concentrations exemplified above, for example, as average values ​​over a specific period during culture. That is, "culture is performed in a medium containing a certain component at a certain concentration," "a certain component is contained in the medium at a certain concentration during culture," or "the concentration of a certain component in the medium during culture is a certain concentration" may mean that the average concentration of the component in the medium over a specific period during culture falls within the range of concentrations. The "average concentration of a certain component in the medium over a specific period during culture" is not particularly limited as long as it allows for understanding fluctuations in the concentration of the component over a specific period during culture. For example, it may mean the average concentration of the component in the medium measured every 60 minutes, every 30 minutes, every 20 minutes, or every 10 minutes over a specific period during culture. The "specific period during culture" may refer to the entire culture period or a portion of the culture period. The "partial culture period" is as described above.

[0118] Each of the various components, such as the active ingredient, may be supplied to the medium throughout the entire culture period, or may be supplied to the medium only during a portion of the culture period. The "part of the culture period" is as described above. Each of the various components, such as the active ingredient, may be supplied to the medium continuously or intermittently, for example. Each of the various components, such as the active ingredient, may be supplied to the medium every day, or every few days, for example.

[0119] The concentrations of various components, such as active ingredients, in the feed medium or perfusion medium may be within the range of the concentrations of the components in the media exemplified above, or may be, for example, 1-fold or more, 1.1-fold or more, 1.3-fold or more, 1.5-fold or more, 2-fold or more, 3-fold or more, 5-fold or more, 7-fold or more, 10-fold or more, 15-fold or more, or 20-fold or more, or 100-fold or less, 70-fold or less, 50-fold or less, 30-fold or less, 20-fold or less, 15-fold or less, 10-fold or less, 7-fold or less, 5-fold or less, 3-fold or less, 2-fold or less, or any combination thereof that is consistent therewith. The concentrations of various components such as active ingredients in the feed medium or perfusion medium may be, for example, 1 to 2 times, 1.1 to 2 times, 1.3 to 2 times, 1.5 to 2 times, 2 to 3 times, 3 to 5 times, 5 to 7 times, 7 to 10 times, 10 to 15 times, 15 to 20 times, 20 to 30 times, 20 to 50 times, 20 to 70 times, or 20 to 100 times the concentrations of the components in the above-exemplified media. The concentrations of various components such as active ingredients in the feed medium or perfusion medium may be, for example, 1 to 100 times, 2 to 50 times, or 5 to 20 times the concentrations of the components in the above-exemplified media.

[0120] The concentrations of any of the components, including the active ingredient, can be measured by known methods used to detect or identify compounds, such as HPLC, UPLC, LC / MS, GC / MS, and NMR.

[0121] By culturing animal cells as described above, muscle tissue (e.g., cultured meat) can be obtained.

[0122] <4> Use of Active Ingredient The present invention discloses the use of the active ingredient in the applications exemplified above. That is, the present invention discloses, for example, the use of the active ingredient for improving muscle quality (e.g., slow-twitch muscle formation) or for obtaining an effect based on improving muscle quality (e.g., slow-twitch muscle formation). The present invention also discloses, for example, the use of the active ingredient for producing a composition for improving muscle quality (e.g., a composition for slow-twitch muscle formation) or for producing a composition for obtaining an effect based on improving muscle quality (e.g., slow-twitch muscle formation).

[0123] The present invention also discloses active ingredients for use in the above-exemplified applications. That is, the present invention discloses, for example, active ingredients for use in improving muscle quality (e.g., slow-twitch muscle formation) and active ingredients for use in obtaining effects based on improving muscle quality (e.g., slow-twitch muscle formation). The present invention also discloses, for example, active ingredients for use in producing a composition for improving muscle quality (e.g., a composition for slow-twitch muscle formation) and active ingredients for use in producing a composition for obtaining effects based on improving muscle quality (e.g., slow-twitch muscle formation).

[0124] The present invention will now be described in more detail with reference to the following non-limiting examples.

[0125] Example 1 Evaluation of the muscle quality improving effect of citicoline or αGPC in vitro using mouse cells In this example, citicoline or αGPC was exposed to a C2C12 cell line differentiated into myotubes, and the muscle quality improving effect of citicoline or αGPC was evaluated using changes in the expression of the Myosin heavy chain 7 (Myh7) gene as an index.

[0126] Induction of differentiation into myotubes: The C2C12 cell line (DS Pharma Biomedical Co., Ltd.), a mouse C3H skeletal muscle myoblast line, was used as myoblasts. The myoblasts were seeded on a plate with a culture area of ​​3.8 cm. 2 12-well plate, 0.16 x 10 per well 5Myoblasts were seeded onto the cells. They were cultured at 37°C in the presence of 5 (v / v)% carbon dioxide. After confirming that the cells had proliferated to approximately 90% of the seeding area, the medium was changed to a differentiation-inducing medium (DMEM medium containing 2 (w / v)% horse serum, 1 (w / v)% penicillin-streptomycin, and 1 (w / v)% glutamine) to induce differentiation into myotubes.

[0127] Evaluation of Myh7 gene expression enhancement by citicoline or αGPC. After confirming myotube differentiation, citicoline or αGPC was added to the differentiation-inducing medium at a concentration of 1 mM. Cells were harvested 24, 48, and 72 hours after addition of citicoline or αGPC, RNA was extracted, and cyclophilin A and Myh7 gene expression was confirmed using real-time quantitative PCR. As a control, cells were harvested from culture medium without citicoline or αGPC and the expression of cyclophilin A and Myh7 genes was confirmed. The cyclophilin A gene was used as an internal control gene. The Myh7 gene is highly expressed in slow-twitch muscle fibers and was used as an indicator of muscle slow-twitch muscle development. Primers for the cyclophilin A gene (SEQ ID NO: 1 and 2) were used. Primers for the Myh7 gene (SEQ ID NO: 3 and 4) were used. The expression level of the Myh7 gene was normalized to the expression level of the cyclophilin A gene.

[0128] Results The results are shown in Figures 1 and 2. In the figures, the expression levels of the Myh7 gene are shown relative to the control group, with the Myh7 gene expression level set at 1 for 24, 48, and 72 hours after the addition of citicoline or αGPC. 72 hours after the addition of 1 mM citicoline, Myh7 gene expression in myotubes was significantly elevated (Figure 1). Furthermore, 48 and 72 hours after the addition of 1 mM αGPC, Myh7 gene expression in myotubes was significantly elevated (Figure 2). These results suggest that both citicoline and αGPC have the effect of improving muscle quality (specifically, slowing down muscle fibers).

[0129] Example 2: Evaluation of the Muscle Quality-Improvement Effect of αGPC in Vivo Using Mice. Using C57BL / 6J mice, the effects of αGPC on changes in muscle quality and muscle function induced by reducing the cage size and restricting activity were examined. Mice were divided into groups to ensure uniform body weight, grip strength, and fat mass. Wide or narrow cages were used. Wide cages measured 220 mm wide x 320 mm long x 165 mm high, while narrow cages measured 110 mm wide x 160 mm long x 130 mm high. The experimental diets were fed to the control group housed in the wide cages, which received a diet containing 20% ​​casein and 7% soybean oil. The groups housed in the narrow cages received either a diet containing 20% ​​casein and 7% lard or a diet containing 20% ​​casein, 7% lard, and 0.2% αGPC. The narrow cage groups were divided into two groups: those with and without exercise using a running wheel. The group configurations are shown in Table 1.

[0130]

[0131] Muscle strength measurement Muscle strength is measured using an animal grip strength measuring device (Chatillon) (Figure 3). The mouse's forelimbs are held in the area from the end of the handle of the grip strength measuring device, and the experimenter slowly pulls the mouse's tail until the mouse lets go. The number at which the mouse lets go is recorded for five consecutive times, and the average of these is calculated as the individual's muscle strength.

[0132] Confirmation of slow-twitch muscle in mouse muscles (1) Confirmation of expression of Myh7 gene, which is highly expressed in slow-twitch muscle fibers, and quantification of mitochondrial DNA. The expression level of the slow-twitch muscle fiber type gene (Myh7) and the amount of mitochondrial DNA present in the gastrocnemius and soleus muscles of mice were confirmed using real-time quantitative PCR.

[0133] (2) Confirmation of Myh7 protein expression, which is highly expressed in slow-twitch muscle fibers. Proteins were extracted from mouse gastrocnemius and soleus muscles and quantified using Western blotting. Specifically, proteins were extracted using RIPA (Radio-Immunoprecipitation Assay) buffer, and the sample adjusted to a protein concentration of 12 μg / 10 μL was used for electrophoresis. After electrophoresis, the sample was transferred to a membrane, and GAPDH and Myh7 proteins were labeled with antibodies. The expression levels of GAPDH and Myh7 proteins were measured using chemiluminescence imaging. The expression level of Myh7 protein was normalized to that of GAPDH protein.

[0134] (3) Confirmation of changes in the number of slow-twitch muscle fibers Frozen tissue sections of mouse gastrocnemius and soleus muscles were prepared, and muscle fiber types were visualized using antibodies against slow-twitch and fast-twitch muscle fibers. The number of muscle fiber types in the prepared sections was counted, and the proportion of slow-twitch muscle fiber types was calculated.

[0135] Confirm improvement in blood sugar levels: Measure diurnal fluctuations in blood sugar levels or blood HbA1c.

[0136] Confirmation of prevention of vascular disorders Measure the expression of genes (ICAM, VCAM, MCP-1, E-selectin) that are markers of vascular disorders (e.g., arteriosclerosis) in the arterial arcuate artery.

[0137] Measurement of body temperature The body surface temperature of the mice is measured during the test period.

[0138] Results: It is expected that administration of αGPC in the non-exercise group will result in increased muscle strength, slow-twitch muscle contraction, increased mitochondrial mass, decreased blood glucose levels or HbA1c, decreased expression of vascular disorder markers, and / or increased body temperature. In other words, it is expected that αGPC will cause slow-twitch muscle contraction in skeletal muscle, thereby improving metabolism. Furthermore, it is expected that improved metabolism will improve blood glucose levels, prevent vascular disorders, and / or increase body temperature.

[0139] In the exercise group, it is expected that exercise-induced increases in muscle strength, slow-twitch muscle contraction, increased mitochondrial mass, decreased blood glucose levels or HbA1c, decreased expression of vascular disorder markers, and / or increased body temperature will be confirmed, and that administration of αGPC will enhance these effects. In other words, it is expected that αGPC will promote exercise-induced slow-twitch muscle contraction, thereby enhancing the metabolic improvement effect of exercise. Furthermore, it is expected that the enhancement of metabolic improvement caused by exercise by αGPC will enhance the blood glucose improvement effect, vascular disorder prevention effect, and / or body temperature increase effect of exercise.

[0140] Example 3: Evaluation of the Muscle Quality-Improvement Effect of Citicoline in Vivo in Mice. The effects of citicoline on muscle quality and function changes induced by reducing the cage size and restricting activity for 30 days were examined using C57BL / 6J mice. Mice were divided into groups with uniform body weight, grip strength, and fat mass. Wide or narrow cages were used. Wide cages measured 220 mm wide x 320 mm long x 165 mm high, while narrow cages measured 110 mm wide x 160 mm long x 130 mm high. The experimental diets were a control group housed in wide cages, fed a diet containing 20% ​​casein and 7% soybean oil, and a diet containing 20% ​​casein and 7% lard or a diet containing 20% ​​casein, 7% lard, and 0.3% citicoline. The narrow cage groups were divided into two groups: one with and one without exercise using a running wheel. The group composition is shown in Table 2. The wheel exercise was performed for five consecutive days, with two days off.

[0141]

[0142] Muscle strength was measured on days 2 and 8 after the start of feeding the donated diet. Muscle strength was measured using an animal grip strength measuring device (Chatillon) (Figure 3). The mouse's forelimbs were held in the grip strength measuring device's handle, which was one-third of the way up, and the experimenter slowly pulled the mouse's tail until the mouse released its grip. The number at which the mouse released its grip was recorded five consecutive times, and the average of these was calculated as the individual's muscle strength.

[0143] Results The results are shown in Figures 4 and 5. On both days 2 and 8 after the start of donor diet feeding, the muscle strength of mice (Group 2) whose cages were narrowed and whose activity levels were restricted for 30 days (Group 1) was reduced compared to control mice (Group 1). Furthermore, on both days 2 and 8 after the start of donor diet feeding, the muscle strength of mice (Group 3) who were subjected to a treadmill exercise program while their activity levels were restricted tended to be higher compared to Group 2. Furthermore, on both days 2 and 8 after the start of donor diet feeding, the muscle strength of mice (Group 4) who were subjected to a treadmill exercise program while their activity levels were restricted and who were given citicoline was significantly higher compared to Group 2. These results suggest that the muscle strength-enhancing effect of exercise is further enhanced by the administration of citicoline.

[0144] Example 4 Evaluation of the muscle quality-improving effect of serine in vitro using mouse cells In this example, serine was exposed to a myofibroblastic C2C12 cell line, and the muscle quality-improving effect of serine was evaluated using changes in the expression of the myosin heavy chain 7 (Myh7) gene as an indicator.

[0145] Evaluation of serine-induced enhancement of Myh7 gene expression. The mouse C3H skeletal muscle myoblast line, C2C12 (DS Pharma Biomedical Co., Ltd.), was used as myoblasts. The myoblasts were seeded on a plate with a culture area of ​​3.8 cm. 2 12-well plate, 0.16 x 10 per well 5Myoblasts were seeded onto the cells. They were cultured at 37°C in the presence of 5% (v / v) carbon dioxide. After confirming cell proliferation at approximately 90% per seeding area, stimulation medium was added. Because serine is naturally contained in DMEM medium, the stimulation medium was prepared by diluting the amino acid concentration of the medium by 1:5 and adding serine to a final concentration of 1, 3, or 10 mM. Cells were harvested 24 hours after addition of the stimulation medium, RNA was extracted, and the expression of cyclophilin A and Myh7 genes was confirmed using real-time quantitative PCR. As a control, cells were harvested from a sample supplemented with DMEM medium diluted by 1:5 instead of the stimulation medium, and the expression of cyclophilin A and Myh7 genes was confirmed. The cyclophilin A gene was used as an internal control gene. The Myh7 gene is highly expressed in slow-twitch muscle fibers and was used as an indicator of muscle slow-twitch muscle development. Primers with sequence numbers 1 and 2 were used for cyclophilin A gene expression. The primers used for the Myh7 gene were those of SEQ ID NOs: 3 and 4. The expression level of the Myh7 gene was normalized with the expression level of the cyclophilin A gene.

[0146] Results The results are shown in Figure 6. In the figure, the expression level of the Myh7 gene is shown as a relative value, with the Myh7 gene expression level in the control group set at 1. 24 hours after serine addition, Myh7 gene expression increased in a serine concentration-dependent manner. In particular, 24 hours after the addition of 10 mM serine, a significant increase in Myh7 gene expression was observed compared to the control group. This suggests that serine has the effect of improving muscle quality (specifically, converting muscles into slow-twitch muscles).

[0147] Example 5: Evaluation of the Muscle Quality-Improving Effect of αGPC in Vivo Using Mice. The effects of αGPC on muscle quality and function changes induced by restricting activity and reducing cage space were examined using C57BL / 6J mice. Mice were divided into groups to ensure uniform body weight, grip strength, and fat mass. Wide or narrow cages were used. Wide cages measured 220 mm wide x 320 mm long x 165 mm high, while narrow cages measured 110 mm wide x 160 mm long x 130 mm high. The experimental diets were a control group housed in wide cages, fed a diet containing 20% ​​casein and 7% soybean oil, and a diet containing 20% ​​casein and 7% lard or a diet containing 20% ​​casein, 7% lard, and 1% αGPC. The narrow cage group was divided into two groups: one with and one without exercise using a running wheel. The group composition is shown in Table 3.

[0148]

[0149] Body composition measurements were performed using the EchoMRI-100 body composition analyzer (Nippon Raytec Corporation) to measure fat mass and muscle mass per body weight.

[0150] The results of fat mass measurements per body weight are shown in Figures 7 and 8. Compared with Group 2, all groups except Group 2 maintained a lower percentage of body fat per body weight (Figure 7). On Day 26 of diet feeding, Group 3 (aGPC-only) also had a lower percentage of body fat per body weight compared with Group 2, and exercise intensity-dependent fat mass was reduced in Groups 4 (3m / min exercise only) and 6 (6m / min exercise only) (Figure 8). Furthermore, on Day 26 of diet feeding, the combination of exercise and aGPC further reduced fat mass in an exercise intensity-dependent manner, as shown in Groups 5 (3m / min exercise + aGPC) and 7 (6m / min exercise + aGPC) (Figure 8).

[0151] The results of muscle weight measurements per unit of body weight are shown in Figures 9 and 10. Compared with Group 2, all groups except Group 2 maintained a higher muscle mass per unit of body weight (Figure 9). On Day 26 of diet feeding, Group 3 (aGPC-only) also had a higher muscle mass per unit of body weight compared with Group 2, and Groups 4 (3 m / min exercise only) and 6 (6 m / min exercise only) showed exercise intensity-dependent increases in muscle weight (Figure 10). Furthermore, on Day 26 of diet feeding, the combination of exercise and aGPC further increased muscle mass in an exercise intensity-dependent manner, as shown in Group 5 (3 m / min exercise + aGPC) and Group 7 (6 m / min exercise + aGPC) (Figure 10).

[0152] Both fat mass reduction and muscle mass increase are indicators of improved body composition. Furthermore, both fat mass reduction and muscle mass increase can also be indicators of toning. These results demonstrate that aGPC administration can improve body composition and tone the body, and that combining aGPC administration with exercise further enhances these effects.

[0153] Measurement of citrate synthase (CS) activity On the final day of the study (day 26 after the start of diet feeding), the gastrocnemius muscles were collected and citrate synthase (CS) activity in the gastrocnemius muscles was measured using the Citrate Synthase Assay Kit ab239712 (Abcam). Citrate synthase (CS) is involved in mitochondrial ATP production.

[0154] The results are shown in Figure 11. Compared with Group 2, CS activity increased in Group 3 (aGPC administration only), Group 5 (3 m / min exercise + aGPC), and Group 7 (6 m / min exercise + aGPC). On the other hand, compared with Group 2, no change in CS activity was observed in Group 4 (3 m / min exercise only) and Group 6 (6 m / min exercise only).

[0155] Increased CS activity is an indicator of increased mitochondrial mass. Increased CS activity can also be an indicator of improved endurance, metabolism, and basic fitness. In other words, the above results show that administration of aGPC has the effect of increasing mitochondrial mass, improving endurance, metabolism, and basic fitness.

[0156] Muscle strength measurements were performed on days 4 and 25 after the start of feeding the provided diet. Muscle strength was measured using an animal grip strength measuring device (Chatillon) (Figure 3). The mouse's forelimbs were held in the area from the end of the handle of the grip strength measuring device, and the experimenter slowly pulled the mouse's tail until the mouse released the grip. The number at which the mouse released the grip was recorded for five consecutive times, and the average of these was calculated as the individual's muscle strength.

[0157] The results are shown in Figures 12-14. Compared with Group 2, Group 7 (6 m / min exercise + aGPC) maintained higher muscle strength from Day 4 onward (Figure 12). Furthermore, Groups 5 (3 m / min exercise + aGPC) and 6 (6 m / min exercise only) showed increased muscle strength from Day 11 onward (Figure 12). Furthermore, Group 3 (aGPC only) showed increased muscle strength from Day 18 onward (Figure 12). Furthermore, Group 7 (6 m / min exercise + aGPC) maintained high muscle strength from Days 4 to 25 onward (Figure 14). Furthermore, on Day 25 onward, muscle strength increased in Groups 3 (aGPC only), 5 (3 m / min exercise + aGPC), 6 (6 m / min exercise only), and 7 (6 m / min exercise + aGPC) compared with Group 2 (Figure 14).

[0158] The above results demonstrate that administration of aGPC can improve muscle strength, and that combining aGPC with exercise can improve muscle strength early. In particular, the combination of 6m / min exercise with aGPC maintained high muscle strength. Furthermore, improved muscle strength can also be an indicator of improved balance, training, rehabilitation, and walking speed. In other words, administration of aGPC alone or in combination with exercise is expected to improve muscle strength, balance, training, rehabilitation, and walking speed.

[0159] Regulating Body Temperature in Subjects Body fat and mitochondrial inner membrane proteins (UCPs) are closely involved in the regulation of lipid metabolism (Edward T, et al. New Advances in Adaptive Thermogenesis: UCP1 and Beyond (2019). Cell Metabolism 29, 27-37.). Furthermore, among UCPs, UCP1, present in brown adipose tissue, regulates body temperature through the regulation of reactive oxygen species (Saijo S, et al. Nardilysin in adipocytes regulates UCP1 expression and body temperature homeostasis (2022). Scientific Reports 12, 3449.). Based on these reports, it is possible that the improvement in body fat caused by αGPC is accompanied by an increase in UCP expression. In particular, it is speculated that an increase in UCP1 expression in brown adipose tissue may have the effect of regulating body temperature through the regulation of reactive oxygen species. Additionally, total energy metabolism and total energy expenditure divided by basal metabolic rate (PAL) are closely related to skeletal muscle mass (Itokawa Yoshinori et al., General Theory of Nutrition, 3rd Revised Edition, Nanzando (2006) 141-164; Susan B, et al., Energy Requirements and Aging (2005). Public Health Nutrition: 8, 1028-1036). It is presumed that the increase in skeletal muscle mass due to αGPC administration increases total energy expenditure and PAL, thereby regulating the body's heat production (body temperature).

[0160] Effect on regulating blood sugar levels in subjects Improving muscle quality leads to an improvement in muscle insulin resistance (Karla E, et al. Role of Skeletal Muscle in Insulin Resistance and Glucose Uptake (2021). Compr Physiol. 10, 785-809.). Improving muscle insulin resistance normalizes glucose uptake in muscles. From these reports, it is inferred that improving muscle quality through αGPC administration can have the effect of regulating blood sugar levels.

[0161] The effect of regulating blood lipid levels in subjects The improvement in body fat due to αGPC administration is thought to be the result of the suppression of body fat accumulation due to improved lipid metabolism. Therefore, it is speculated that αGPC administration has the effect of improving lipid clearance in the blood.

[0162] According to the present invention, a technique for improving muscle quality and related techniques can be provided.

[0163] <Explanation of Sequence Listing> SEQ ID NOS: 1 to 4: Primers

Claims

1. A composition for improving muscle quality, containing a choline-based compound.

2. A composition containing a cholinergic compound for obtaining effects based on improving muscle quality.

3. The composition according to claim 2, wherein the effect based on improving muscle quality is one or more effects selected from the group consisting of an effect of increasing the amount of mitochondria in muscle, an effect of improving body composition, an effect of improving endurance, an effect of improving metabolism, an effect of improving balance ability, an effect of improving muscle strength, an effect of improving walking speed, an effect of toning the body, an effect of improving basic physical fitness, an effect of improving training effects, an effect of improving rehabilitation effects, an effect of regulating the body temperature of a subject, an effect of regulating the blood glucose level of a subject, an effect of regulating the blood lipid concentration of a subject, and an effect of preventing, improving, and / or treating undesirable symptoms related to muscles.

4. The composition of claim 3, wherein the symptom is one or more symptoms selected from the group consisting of disuse muscle atrophy, neurogenic muscle atrophy, myogenic muscle atrophy, myasthenia gravis, cachexia, frailty, musculoskeletal syndrome, metabolic syndrome, decreased basic physical strength, decreased walking speed, lower back pain, stiff shoulders, headache, hyperglycemia, hyperlipidemia, vascular disorder, hypothermia, and sensitivity to cold.

5. The composition according to any one of claims 1 to 4, wherein the improvement of muscle quality is slow-twitch muscle formation.

6. The composition according to any one of claims 1 to 4, wherein the choline-based compound is one or more components selected from the group consisting of sn-glycero-3-phosphocholine, citicoline, choline, choline phosphate, phosphocholine, phosphatidylcholine, sphingomyelin, and serine.

7. The composition according to any one of claims 1 to 4, wherein the choline-based compound is one or more components selected from the group consisting of sn-glycero-3-phosphocholine, citicoline, and serine.

8. The composition according to any one of claims 1 to 4, which is a pharmaceutical composition.

9. The composition according to any one of claims 1 to 4, which is a food composition.

10. The method according to any one of claims 1 to 4, wherein the content of the choline compound is 100 ppm (w / w) to 99.9% (w / w). The composition according to any one of claims 1 to 4.

11. The composition according to any one of claims 1 to 4, which is used in conjunction with performing physical exercise.

12. The composition according to any one of claims 1 to 4, wherein when administered in combination with physical exercise, muscle strength improves earlier in a subject to which the composition is administered compared to when the composition is not administered in combination with physical exercise.

13. A method for improving muscle quality in a subject, comprising administering a cholinergic compound to the subject.

14. A method for obtaining an effect based on improving muscle quality in a subject, comprising a step of administering a cholinergic compound to the subject.

15. The method of claim 14, wherein the effect based on improving muscle quality is one or more effects selected from the group consisting of an effect of increasing the amount of mitochondria in muscle, an effect of improving body composition, an effect of improving endurance, an effect of improving metabolism, an effect of improving balance ability, an effect of improving muscle strength, an effect of improving walking speed, an effect of toning the body, an effect of improving basic physical fitness, an effect of improving training effects, an effect of improving rehabilitation effects, an effect of regulating the body temperature of a subject, an effect of regulating the blood glucose level of a subject, an effect of regulating the blood lipid concentration of a subject, and an effect of preventing, improving, and / or treating undesirable symptoms related to muscles.

16. The method of claim 15, wherein the symptom is one or more symptoms selected from the group consisting of disuse muscle atrophy, neurogenic muscle atrophy, myogenic muscle atrophy, myasthenia gravis, cachexia, frailty, musculoskeletal syndrome, metabolic syndrome, decreased basic physical strength, decreased walking speed, lower back pain, stiff shoulders, headache, hyperglycemia, hyperlipidemia, vascular disorder, hypothermia, and sensitivity to cold.

17. The dosage of the cholinergic compound according to any one of claims 13 to 16 is 0.01 mg / kg body weight / day or more.

10. The method according to any one of claims 1 to 9.

18. The method according to any one of claims 13 to 16, wherein the dosage of the cholinergic compound is 1000 mg / kg body weight / day or less.

10. The method according to any one of claims 1 to 9.

19. The method according to any one of claims 13 to 16, which is used in conjunction with performing physical exercise.

20. The method according to any one of claims 13 to 16, wherein when combined with physical exercise, muscle strength improves earlier in the subject than when combined with no physical exercise.

21. A method for improving muscle quality in cultured meat, comprising culturing animal cells in the presence of a choline-based compound.

22. A method for producing cultured meat, comprising culturing animal cells in the presence of a choline-based compound.

23. 23. The method of claim 22, wherein the cultured meat produced is cultured meat with improved muscle quality.

24. The method according to any one of claims 13 to 16, wherein the muscle quality improvement is muscle slow-twitching.

25. 25. The method of claim 24, wherein the slow-twitch muscle fiber conversion improves the texture of the cultured meat and / or the flavor of the cultured meat.

26. 26. The method of claim 25, wherein the improved texture is improved water retention of the cultured meat and / or improved juiciness of the cultured meat.

27. The method of claim 25, wherein the improved flavor is an improvement in the taste of the cultured meat due to an increase in the amount of free amino acids.

28. The method according to any one of claims 13 to 16, wherein the choline-based compound is one or more components selected from the group consisting of sn-glycero-3-phosphocholine, citicoline, choline, choline phosphate, phosphocholine, phosphatidylcholine, sphingomyelin, and serine.

29. The method according to any one of claims 13 to 16, wherein the choline-based compound is one or more components selected from the group consisting of sn-glycero-3-phosphocholine, citicoline, and serine.