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WO2019026930A1 - Food composition for increasing muscle mass, muscle mass increasing agent, therapeutic agent for locomotive syndrome, therapeutic agent for sarcopenia, exercise function maintenance agent, muscle atrophy-related gene expression inhibitor, muscle protein degradation inhibitor, muscle atrophy prevention agent, muscle synthesis-related gene expression promoter, and muscle synthesis promoter - Google Patents

Food composition for increasing muscle mass, muscle mass increasing agent, therapeutic agent for locomotive syndrome, therapeutic agent for sarcopenia, exercise function maintenance agent, muscle atrophy-related gene expression inhibitor, muscle protein degradation inhibitor, muscle atrophy prevention agent, muscle synthesis-related gene expression promoter, and muscle synthesis promoter Download PDF

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Publication number
WO2019026930A1
WO2019026930A1 PCT/JP2018/028756 JP2018028756W WO2019026930A1 WO 2019026930 A1 WO2019026930 A1 WO 2019026930A1 JP 2018028756 W JP2018028756 W JP 2018028756W WO 2019026930 A1 WO2019026930 A1 WO 2019026930A1
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WIPO (PCT)
Prior art keywords
muscle
euglena
agent
related gene
active ingredient
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Ceased
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PCT/JP2018/028756
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French (fr)
Japanese (ja)
Inventor
綾香 中島
鈴木 健吾
立花 宏文
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kyushu University NUC
Euglena Co Ltd
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Kyushu University NUC
Euglena Co Ltd
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Priority claimed from JP2017254319A external-priority patent/JP6524204B2/en
Application filed by Kyushu University NUC, Euglena Co Ltd filed Critical Kyushu University NUC
Priority to CN201880049823.XA priority Critical patent/CN111031812A/en
Publication of WO2019026930A1 publication Critical patent/WO2019026930A1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/02Algae
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00Drugs for disorders of the muscular or neuromuscular system

Definitions

  • the present invention provides a food composition for increasing muscle mass, an agent for increasing muscle mass, an agent for treating locomotive syndrome, an agent for treating sarcopenia and a maintenance agent for motor function, an agent for suppressing muscle atrophy-related gene expression, an agent for suppressing muscle protein degradation, an agent for preventing muscle atrophy
  • the present invention relates to a muscle synthesis related gene expression promoter and a muscle synthesis promoter.
  • Non-Patent Document 1 According to the Japanese Orthopedic Association, locomotive syndrome is a disorder in one or more of bone, joints, cartilage, intervertebral disc, muscles such as intervertebral disc, muscles, and functions such as "stand up” or "walk” are reduced. It is defined that it is a state (nonpatent literature 2).
  • Sarcopenia refers to muscle weakness associated with loss of muscle mass due to aging, illness, lack of exercise, malnutrition, etc., and decrease in physical function
  • locomotive syndrome refers to exercise of muscles, bones, etc. due to aging or lack of exercise It points to the state where troubles occurred in the part of the vessel and daily life became difficult.
  • locomotive syndrome is a concept that includes general motor symptoms
  • sarcopenia can be said to be a symptom that focuses on muscle mass, muscle strength, and physical function among the motor organs.
  • Euglena (genus: Euglena, Japanese name: Midorimusi) is attracting attention as a biological resource which is considered to be promising for food, feed, fuel and the like.
  • Euglena is a supplement for the well-balanced intake of many types of nutrients, with 59 types of nutrients corresponding to most of the nutrients necessary for human beings to live including vitamins, minerals, amino acids, unsaturated fatty acids, etc. It has been suggested that it could be used as a food source in poor areas where it can not consume necessary nutrients.
  • Euglena is a primary producer of the food chain, and it is a large-scale culture because it is difficult to prey on predators and culture conditions such as light, temperature conditions, and agitation speed are higher than other microorganisms.
  • our extensive research has established a large-scale culture technique and opened the way to large-scale supply of Euglena-derived substances such as Euglena and Paramylon extracted from Euglena.
  • Euglena is a unique organism that has the property of carrying out flagellum movement and at the same time carries out photosynthesis with chloroplast as a plant, and it is expected that Euglena itself and substances derived from Euglena have many functions. ing. Therefore, it is desirable to develop a method of using Euglena and Euglena-derived substances that can be supplied in large quantities.
  • the present invention has been made in view of the above problems, and the object of the present invention is to be able to be taken continuously continuously, to effectively increase muscle mass, to treat locomotive syndrome and sarcopenia, Food composition for increasing muscle mass that can be prevented and improved, muscle mass increasing agent, locomotive syndrome therapeutic agent, sarcopenia therapeutic agent, motor function maintenance agent, muscle atrophy related gene expression inhibitor, muscle protein degradation inhibitor, An object of the present invention is to provide a muscle atrophy preventive agent, a muscle synthesis related gene expression promoter and a muscle synthesis promoter.
  • the present inventors have an effect of increasing muscle mass in Euglena, an action of suppressing muscle atrophy, an action of suppressing degradation of muscle proteins, and an action of promoting muscle synthesis.
  • the present invention has been completed.
  • the above-mentioned subject is solved by the food composition for increasing muscle mass characterized by containing an Euglena-derived substance as an active ingredient.
  • the Euglena-derived substance is preferably Euglena.
  • it may be a muscle mass increasing food composition used to increase the amount of quadriceps.
  • the above-mentioned problems are solved by a muscle mass increasing agent characterized by containing an Euglena-derived substance as an active ingredient.
  • a food composition containing an Euglena-derived substance as an active ingredient for sarcopenia prevention, sarcopenia improvement, locomotive syndrome prevention or locomotive syndrome improvement.
  • a locomotive syndrome preventive agent, a locomotive syndrome therapeutic agent, a sarcopenia preventive agent, a sarcopenia therapeutic agent, a motor function maintenance agent exercise comprising the euglena-derived substance as an active ingredient. It is solved by the function improving agent.
  • the subject contains an Euglena-derived substance as an active ingredient, and is for suppressing muscle atrophy-related gene expression, for suppressing muscle protein degradation, for preventing muscle atrophy, for promoting muscle synthesis-related gene expression or It is solved by a food composition for promoting synthesis.
  • a muscle atrophy-related gene expression inhibitor characterized by containing an Euglena-derived substance as an active ingredient, a muscle protein degradation inhibitor, a muscle atrophy preventive agent, a muscle synthesis related gene expression It is solved by the promoter and the muscle synthesis promoter.
  • the agent for preventing atrophy, muscle synthesis related gene expression promoter and muscle synthesis promoter can be provided.
  • the synthetic related gene expression promoter and muscle synthesis promoter have no reports of adverse effects and have Euglena as the active ingredient with a level of safety that conforms to the Food Sanitation Law. It can be taken.
  • Experiment 1 it is a graph which shows a body weight change when each test meal is continuously fed for 4 weeks.
  • 6 is a graph showing the amount of food consumed by the test food in Test 1. It is a graph which shows the liver weight at the end of test 1. It is a graph which shows quadriceps femoris muscle weight at the end of test 1 (Tukey's test, * p ⁇ 0.05 vs. control group).
  • a muscle mass increasing food composition containing an Euglena-derived substance as an active ingredient, a muscle mass increasing agent, a locomotive syndrome therapeutic agent, a sarcopenia therapeutic agent, a motor function maintenance agent, a muscle atrophy-related gene expression inhibitor, a muscle
  • the present invention relates to a protein degradation inhibitor, a muscle atrophy preventive agent, a muscle synthesis related gene expression promoter and a muscle synthesis promoter.
  • locomotive syndrome (locomotive syndrome) is characterized by the ability to “stand up” or “walk” due to injury to one or more of bone, joint, cartilage, intervertebral disc, muscle, or muscle It is defined as being in a state of decline (Non-Patent Document 2).
  • Locomotive syndrome may be abbreviated as Locomo, or may be called locomotive syndrome, motor organ syndrome or the like.
  • diagnostic criteria for locomotive syndrome will be described (Noriko Yoshimura, “Clinical diagnostic value and prevalence of locomotive syndrome", Japanese Geriatrics Journal 52, 4 350-353 (2015)).
  • the locomotion test which is one of diagnostic criteria for locomotive syndrome, comprises (1) rising test, (2) two-step test, and (3) locomo 25.
  • Stand-up test is a test of measuring the leg strength based on whether one leg or both legs can stand up by preparing a stand of four heights of 10 cm, 20 cm, 30 cm and 40 cm.
  • Second In the two-step test walk two steps with as much as possible, measure the stride for two steps, and divide by the height to calculate a two-step value. Evaluate walking ability including leg strength, balance ability, flexibility etc. of lower limbs by 2-step value.
  • Locomo 25 evaluates with a 25-item questionnaire about whether or not there was body pain or difficulty in daily life in the past one month.
  • locomotion degree 1 The clinical judgment value of locomotion degree 1 is as follows. 1) Rise test: Can not get up from a height of 40 cm with either one leg 2) 2 step test: 2 step value less than 1.3 3) Locomo 25: Locomo 25 result is 7 points or more Above 1)- If any one of the above 3) applies, it is determined as "Locomor 1", and it is determined that the mobile function has started to deteriorate.
  • locomotion degree 2 The clinical judgment value of locomotion degree 2 is as follows. 1) Rise test: Can not stand from height of 20 cm with both legs 2) 2 step test: 2 step value less than 1.1 3) Locomo 25: Locomo 25 result is 16 points or more Any of the above 1) to 3) If any one of the cases is true, it is judged as "Locomor 2", and it is judged that the deterioration of the mobile function is progressing.
  • Sarcopenia is a syndrome characterized by atrophy of skeletal muscle with age, accompanied by loss of skeletal muscle mass and muscle strength or loss of physical function.
  • sarcopenia many diagnostic criteria that differ depending on the country, region, etc. have been proposed, but it is considered as a sarcopenia when muscle loss is an essential item, and it falls under either muscle strength or physical ability "Sarcopenia's diagnosis, pathophysiology and treatment" Journal of Japan Geriatrics Journal Vol. 52, No. 4, 343-349 (2015)).
  • the limb skeletal muscle mass is the muscle mass of the limb divided by the height (m) squared
  • DXA dual energy X-ray absorptiometry
  • the walking speed of 10m is less than 0.8 m / sec, or grip strength in men Sarcopenia is diagnosed if it is less than 30 kg and less than 20 kg in women.
  • the procedure of diagnosis is to measure the walking speed first and measure limb skeletal muscle mass if it is less than 0.8 m / s, and it is 7.23 kg / m 2 or less for men and 5.67 kg / m for women. If it is m 2 or less, sarcopenia is diagnosed. If it is high, it is not diagnosed as sarcopenia. If walking speed is faster than 0.8 m / sec, grip strength is measured and if it is higher than 30 kg for males and 20 kg for females it is not diagnosed as sarcopenia, if it is higher, limb skeletal muscle mass is measured Sarcopenia if below the diagnostic threshold, not sarcopenia if above the diagnostic threshold.
  • the diagnostic criteria of the EWGSOP are the criteria developed by the working group in Europe, and because there is a difference in physique and physical function between Europeans and Asians, the working group for sarcopenia in Asia, ASIAN working Group FOR SARCPOPENIA (AWGS) Has established diagnostic criteria for Asians.
  • the AWGS diagnostic criteria differ from the EWGSOP diagnostic criteria and the criteria for grip strength and muscle mass.
  • the standard of grip strength diagnosed with sarcopenia is less than 26 kg for men and less than 18 kg for women, and the criteria for muscle mass are 7 kg / m 2 or less for men (DXA value, BIA value), 5.4 kg for women 2 or less (DXA value), 5.7 kg / m 2 or less (BIA value).
  • the diagnostic criteria in the EWGSOP are based on the elderly of Western Americans, and Japanese and European Americans have differences in physique and lifestyle even with the elderly, so the simplified criteria of sarcopenia suitable for the elderly of Japanese A draft has been prepared by the National Longevity Medical Research Center Long-Term Longitudinal Epidemiological Study on Aging (NILS-LSA).
  • BMI values body mass index, height
  • a diagnosis of sarcopenia is made if the ratio of body weight to the square is an index representing body size) less than 18.5, or the lower leg circumference is less than 30 cm.
  • Normal when walking speed and grip strength are above standard values. Even if the walking speed and grip strength are below the standard value, BMI and lower leg circumference are above the standard value, it is diagnosed as a vulnerable elderly person but not sarcopenia.
  • Euglena includes all microorganisms classified into Euglena (Euglena) according to the zoological and botanical classification, variants thereof, and variants thereof.
  • the microorganisms of Euglena (Euglena) are, in zoology, the protozoan (Protozoa) dinoflagellate (Mastigophorea), and the Euglenoidina (Euglenida) belonging to the plant dinoflagellate (Phytomastigophorea) It is a microorganism belonging to the eye (Euglenoidina).
  • microorganisms of the genus Euglena belong to the order Euglenales belonging to the family Euglenophyceae of the family Agromorpha (Euglenophyta) in botany.
  • Euglena gracilis E. gracilis
  • E. gracilis E. gracilis
  • E. gracilis Z strain E. gracilis (E. gracilis) NIES-49 strain, etc.
  • Euglena ⁇ Mutant strain SM-ZK of E. gracilis Z strain chloroplast-deficient strain
  • Bacillaris, ⁇ -1 derived from gene mutant strains such as chloroplast mutant strains of these species , 3-glucanase, Euglena intermedia, Euglena piride, and other Euglenas, such as Astaia longa.
  • the genus Euglena is widely distributed in fresh water such as ponds and marshes, and may be used separately from them, or any Euglena isolated previously may be used.
  • the genus Euglena includes all its variants. Further, among these mutant strains, those obtained by genetic methods such as recombination, transduction, transformation and the like are also contained.
  • a culture solution for example, a culture solution to which nutrients such as nitrogen source, phosphorus source, and minerals have been added, for example, modified Cramer-Myers medium ((NH 4 ) 2 HPO 4 1.0 g / L , KH 2 PO 4 1.0 g / L, MgSO 4 ⁇ 7 H 2 O 0.2 g / l, CaCl 2 ⁇ 2 H 2 O 0.02 g / l, Fe 2 (SO 2 ) 3 ⁇ 7 H 2 O 3 mg / l, MnCl 2 ⁇ 4 H 2 O 1.8 mg / l, CoSO 4 ⁇ 7 H 2 O 1.5 mg / l, ZnSO 4 ⁇ 7 H 2 O 0.4 mg / l, Na 2 MoO 4 ⁇ 2 H 2 O 0.2 mg / l, CuSO 4 ⁇ 5H 2 O 0.02g / l, thiamine hydrochloride (vitamin B1) 0.1mg / l, thiamine hydrochloride (vitamin B1)
  • (NH 4 ) 2 HPO 4 can also be converted to (NH 4 ) 2 SO 4 or NH 3 aq.
  • well-known Hutner's medium and Koren-Hutner's medium prepared based on the description of Euglena Physiology and Biochemistry (Kitaoka Shozaburo, ed., Inc. publication center) may be used.
  • the pH of the culture solution is preferably 2 or more, and the upper limit thereof is preferably 6 or less, more preferably 4.5 or less.
  • photosynthetic microorganisms can be more preferentially grown than other microorganisms, so that contamination can be suppressed.
  • the culture of Euglena cells may be performed by an open pond method using sunlight directly, a method of sending sunlight collected by a light collecting device by an optical fiber or the like and irradiating it in a culture tank and utilizing it for photosynthesis.
  • culture of Euglena cells may be performed using, for example, a fed-batch method, but culture using a flask culture or a fermenter, batch culture method, half-batch culture method (feed-batch culture method), continuous culture method (perfusion It may be carried out by any liquid culture method such as culture method).
  • the separation of Euglena cells is performed, for example, by centrifugation, filtration or simple sedimentation of the culture fluid.
  • Euglena live cells separated by centrifugation, filtration, sedimentation or the like can be used as Euglena algal cells as they are.
  • Euglena living cells can be used as they are after harvesting from a culture tank, but washing with water or saline is preferred. Also, it may be used in the form of a dispersion in which Euglena algal cells are dispersed in a liquid such as water.
  • a mechanically treated product of algal cells obtained by performing ultrasonic irradiation treatment or mechanical treatment such as homogenization may be used as the euglena algal cells.
  • “Euglena aqueous solvent extract” means an extract extracted from Euglena using an aqueous solvent, and in particular, using water as an aqueous solvent, several seconds to several seconds at 5 ° C. to 600 ° C. It is preferred to use a water extract or a hot water extract of Euglena extracted for 10 hours.
  • the water used for the extraction does not necessarily have to be distilled water, pure water, or ultrapure water, and may be, for example, tap water or an impurity, but contains a component that hinders the extraction of the active ingredient. No water is preferred.
  • water extract means an extract with water of 0 to 50 ° C. (excluding 0 ° C.).
  • water means water of 0 to 50 ° C. (excluding 0 ° C.).
  • the temperature of water is not particularly limited as long as the active ingredient can be sufficiently extracted without affecting the active ingredient, but preferably 1 to 40 ° C., more preferably 5 to 35 ° C. Particularly preferred is 10 to 30 ° C.
  • hot water extract means an extract with water at a temperature higher than 50 ° C., and can also be called “hot water extract”.
  • hot water means water having a temperature higher than 50 ° C., and is a concept including “hot water”, including water in a boiling state. Further, it is not limited to hot water in a liquid state, and includes hot water in a gaseous state and in a supercritical state.
  • the temperature of the hot water is not particularly limited as long as the active ingredient can be sufficiently extracted without affecting the active ingredient, but is preferably higher than 50 ° C. and 120 ° C. or less, more preferably 50 It is higher than ° C and not higher than 100 ° C.
  • the pH of water used for extraction is not particularly limited as long as the active ingredient can be extracted sufficiently without affecting the active ingredient, but the pH is preferably 4 to 10, more preferably 5 to 9 The pH is particularly preferably 6 to 8.
  • water is used alone as the aqueous solvent, but the active ingredient can be sufficiently extracted without affecting the active ingredient, and one solvent can be generally used for extraction. Or you may select and use 2 or more types.
  • water, alcohol, glycols etc. can be mentioned, it is not limited to this.
  • alcohols include ethanol, methanol, n-propanol and isopropanol.
  • glycols include butylene glycol and propylene glycol.
  • Other aqueous solvents include acetone and the like. These solvents may be used alone or as an aqueous solution, and may be used as any two or more mixed solvents.
  • the temperature of the aqueous solvent used for extraction is, for example, 0 ° C. or higher, and is not particularly limited as long as it does not affect the active ingredient.
  • an aqueous solvent in a boiling or supercritical state can be used, it is preferable to use an aqueous solvent at 5 ° C. to 600 ° C., and it is more preferable to use an aqueous solvent at 10 ° C. to 200 ° C. Therefore, the aqueous solvent for extraction also includes an aqueous solvent in a boiling state or a supercritical state.
  • the amount of aqueous solvent used for extraction is preferably an amount capable of sufficiently dissolving the water-soluble active ingredient contained in Euglena.
  • the extraction method is also not particularly limited, and, for example, the extraction can be performed by the method described below, but it is not limited thereto, and a common extraction method can be freely selected and used.
  • Euglena water extraction can be carried out by the usual method as shown below, but is not limited thereto.
  • Euglena tissue and water are placed in a container and allowed to stand for a predetermined time while stirring or shaking as appropriate, and the obtained extract can be used as it is as a water extract.
  • the supernatant obtained by centrifuging such extract can be used as a water extract.
  • such extract or supernatant may be concentrated, dried to remove water and used as a water extract.
  • the water extraction may be performed by adding a small amount, for example, 10% by mass or less of an alcohol, preferably ethanol, to the water in order to increase the extraction efficiency and shorten the extraction time.
  • the extraction time in the case of water extraction is not particularly limited as long as it is a time during which the active ingredient is extracted, and can be appropriately set according to the temperature of extraction within a range of several seconds to several tens of hours.
  • Extraction with hot water can be carried out by a commonly used method as shown below, but it is not limited thereto. Extraction is carried out by heating Euglana after introducing it into a commonly used extractor together with water. In the case of extraction using boiling water or water in the supercritical state, it is necessary to use an extractor capable of withstanding the water vapor pressure.
  • the pressure at the time of extraction can be set to 1 to 5000 atm, preferably 60 to 400 atm. In the case of extraction under high temperature and pressure, if the extraction time is too long, the active ingredient may be decomposed or a chemical reaction may occur. Therefore, when extraction is performed under high temperature and high pressure, the extraction time is preferably short, for example, 3 minutes or less, more preferably 1 minute or less, and particularly preferably 30 seconds or less.
  • the extracted Euglena extract can be used as it is as an active ingredient of the muscle mass increasing agent according to the present embodiment, the extract can be further separated by an appropriate separation means (eg, distribution extraction, gel filtration method, silica gel chromatography It is also possible to fractionate the highly active fraction by chromatography, reverse phase or normal phase high performance liquid chromatography, etc.).
  • the Euglena extract or a fraction thereof may be concentrated and dried to remove an aqueous solvent, which may be used as an aqueous solvent extract.
  • the muscle mass increasing agent according to the present embodiment is a muscle mass increasing agent (muscle mass increasing agent) containing an Euglena-derived substance as an active ingredient.
  • the “Euglena-derived substance” includes Euglena algal cells such as Euglena live cells and dried algal cells of Euglena, as well as aqueous solvent extracts of Euglena, processed products of Euglena algal bodies, and the like.
  • muscle mass increaser refers to an agent that increases muscle mass (muscle mass) and is synonymous with “muscle mass increaser”.
  • the increase in muscle mass can be evaluated, for example, by measuring the muscle mass using a muscle mass meter or the like.
  • the muscle targeted for increase in muscle mass is not particularly limited, but skeletal muscle is preferred.
  • the skeletal muscle includes, for example, chest-crest mastoid muscle, pectoralis major muscle, minor pectoral muscle, anterior serratus muscle, subclavian muscle, rectus abdominis muscle, external oblique muscle, internal oblique muscle, transverse abdominal muscle, hippocampus muscle, Cap muscles, latissimus dorsi, erector spinae muscles, elevated shoulder muscles, rhomboids, deltoids, detrusors, supraspinatus, infraspinatus, subscapularis, great fornix, brachial muscles, biceps brachii , Brachial muscle, brachial muscle, triceps brachial muscle, elbow muscle, circular pronation muscle, square pronation muscle, pronation muscle, ulnar carpal flexor muscle, dorsal carpal flexor muscle, long palm muscle, superficial flexor muscle
  • the quadriceps muscles are involved in the extension of the knee joint and the flexion of the hip joint, specifically, the action of stretching and standing the knee, the walking action, and the running action. Therefore, when the muscle mass of the quadriceps femoris increases, movements such as standing up, walking and running improve, and locomotive syndrome and sarcopenia can be prevented and improved.
  • the euglena-derived substance can be used as a muscle enhancing agent since it has an action to increase muscle mass.
  • the "muscle-enhancing agent” refers to an agent that strengthens muscles, and muscle strengthening can be evaluated by, for example, muscle mass and muscle strength.
  • “Strengthening muscles” means increasing muscle mass and improving muscle strength. An increase in muscle mass is determined, for example, from measurement of muscle mass, and an improvement in muscle strength is determined, for example, from measurement of muscle strength by a muscle meter or the like.
  • the muscle mass increaser contains an Euglena-derived substance as an active ingredient, and is an agent for preventing locomotive syndrome for preventing or treating locomotive syndrome and sarcopenia, an agent for treating locomotive syndrome, an agent for preventing sarcopenia, and a treatment for sarcopenia Can also be used.
  • “To treat locomotive syndrome” and “to treat sarcopenia” refer to the symptoms of locomotive syndrome and sarcopenia compared to the condition before taking the euglena-derived substance which is the active ingredient of the muscle mass increasing agent of this embodiment. It means to reduce.
  • the muscle mass increaser according to the present embodiment contains an Euglena-derived substance as an active ingredient, and may be used also as a motor function maintenance agent for maintaining the motor function or a motor function improvement agent for improving the motor function. it can. “Maintaining the motor function” means “stand up” or “walk” as compared with the state to be compared, for example, the state before taking the euglena-derived substance which is the active ingredient of the muscle mass increasing agent of the present embodiment. It means that motor function such as is maintained.
  • “Improvement of motor function” means “stand up” or “walk” as compared with the condition to be compared, for example, the condition before taking the euglena-derived substance which is the active ingredient of the muscle mass increasing agent of the present embodiment. Say that motor function such as is improved.
  • the Euglena-derived substance has an action of suppressing the expression of a muscle atrophy-related gene, it can be used as a muscle atrophy-related gene expression inhibitor, a muscle atrophy preventive agent, and a muscle protein degradation inhibitor.
  • muscle atrophy-related genes whose expression is suppressed by euglena-derived substances include, for example, ubiquitin ligase genes (Cbl-b (SEQ ID NO: 1), Atrogin-1 (SEQ ID NO: 2), MuRF1 (SEQ ID NO: 3)) Is illustrated.
  • Muscle synthesis related gene expression promoter and muscle synthesis promoter Euglena-derived substances have the effect of promoting the expression of muscle synthesis-related genes, and thus can be used as muscle synthesis-related gene expression promoters and muscle synthesis promoters.
  • a muscle synthesis related gene whose expression is promoted by the euglena-derived substance, for example, the gene of S6K (SEQ ID NO: 5) which promotes the synthesis of muscle protein is exemplified.
  • the muscle mass increasing agent containing the Euglena-derived substance according to the embodiment as an active ingredient is administered to a patient with decreased muscle mass or muscle strength, or a patient who has a definitive diagnosis of locomotive syndrome or sarcopenia.
  • the muscle mass increasing agent containing the Euglena-derived substance of the present embodiment as an active ingredient is constituted as a food composition such as a pharmaceutical composition, a health food, etc. Prophylactically administered to people with locomotive syndrome or sarcopenia reserve forces, such as those who are likely to suffer from locomotive syndrome or sarcopenia.
  • Euglena-derived substances such as Euglena algae can be taken as food and have no side effects, so they can be administered even before a definitive diagnosis of locomotive syndrome or sarcopenia.
  • the muscle mass increasing food composition containing the Euglena-derived substance according to the present embodiment as an active ingredient, and the target for administering the muscle mass increasing agent are limited to persons having the above condition or condition or animals other than human beings Absent.
  • a muscle composition for increasing muscle mass and an agent for increasing muscle mass which contain an Euglena-derived substance as an active ingredient, can be administered to humans of 40 years of age or older, particularly to elderly people of 60 or 65 years of age. People over the age of 40, especially elderly people over the age of 60 or 65, tend to lose muscle mass and muscle strength with age, but with the Euglena-derived substance, the muscle mass increasing action Can reduce muscle mass loss and muscle strength loss.
  • the food composition for increasing muscle mass and muscle mass increasing agent of the present embodiment since muscle mass can be increased, prevention and treatment of diseases associated with decrease in muscle mass and muscle strength, bed rest state It is expected to be applied to the prevention of injuries and the prevention of injuries caused by falls.
  • the muscle mass increasing agent containing the Euglena-derived substance according to the present embodiment as an active ingredient may be added to a pharmacologically acceptable additive and used as a composition such as a food composition, a pharmaceutical composition, etc. it can.
  • the Euglena-derived substance of the present embodiment can also be used for food.
  • food compositions for preventing, muscle synthesis-related gene expression promotion, or muscle synthesis promotion use various amounts of food materials as an effective amount of an euglena-derived substance capable of effectively exerting the intended action. By blending, it is possible to provide a food composition having the effect.
  • the present invention can provide a food composition of food labeled as for increasing muscle mass and the like in the field of food.
  • the food composition include, in addition to general foods, food for specified health use, nutritive function food, functional display food, food for hospital patients, supplement and the like. It can also be used as a food additive.
  • Examples of the food composition include seasonings, processed meat products, processed agricultural products, beverages (soft drinks, alcoholic drinks, alcoholic drinks, carbonated drinks, milk drinks, fruit drinks, tea, coffee, nutritional drinks, etc.), powdered drinks (powders) Juices, powdered soups, etc., concentrated beverages, confectionery (candy (nodice), cookies, biscuits, gums, gummi, chocolate, etc.), bread, cereals, etc. can be mentioned. Further, in the case of food for specified health use, nutritive function food, functional display food and the like, it may be in the form of capsules, troches, syrups, granules, powders and the like.
  • the food for specified health use is a food containing a health functional ingredient that affects physiological functions etc., and can be displayed as being suitable for a specific use of health with the permission of the Commissioner of the Consumer Agency. is there.
  • the present invention as a specific health application, "helps to suppress the decrease in muscle mass and muscle strength necessary to live an independent daily life”, “helps to improve walking ability”, “maintenance of muscle mass and muscle strength”, “Support for maintenance of muscle mass and muscle strength”, “maintenance of walking ability that declines with age in middle-aged and elderly people”, “Supporting the ability to create muscles that help maintain muscles that decline with aging,” etc.
  • the nutritive function food is a food used for supplementation of nutritional components (vitamins, minerals), and displays the function of the nutritional components.
  • the amount of nutritional component contained in the daily intake standard amount is necessary for the amount of nutritional component contained in the daily intake standard amount to be within the defined upper limit value and lower limit value, and not only the display of nutritional function but also alert display etc.
  • the functional labeling food is a food which displays scientific basis-based functionality at the responsibility of the business operator. Information on the basis of safety and functionality etc. was sent to the Secretary General of the Consumer Agency before sales.
  • the food composition according to the present embodiment in addition to the Euglena-derived substance, it is possible to freely select and mix one or two or more kinds of components that can be used for the food composition.
  • all additives which can usually be used in the food field can be contained, such as various seasonings, preservatives, emulsifiers, stabilizers, flavors, coloring agents, preservatives, pH adjusters and the like.
  • HMB 3-hydroxy-3-methylbutyrate
  • the muscle mass increase agent of the present embodiment locomotive syndrome preventive agent, locomotive syndrome therapeutic agent, sarcopenia preventive agent, sarcopenia therapeutic agent, motor function maintenance agent, motor function improvement agent, muscle atrophy related gene expression inhibitor, muscle protein degradation inhibitor
  • a muscle atrophy preventive agent, a muscle synthesis related gene expression promoter and a muscle synthesis promoter are used together with a pharmaceutically acceptable carrier or additive together with an amount of an euglena-derived substance capable of effectively exerting the intended action.
  • a pharmaceutical composition having the above-mentioned action is provided.
  • the pharmaceutical composition may be a pharmaceutical or quasi-drug.
  • the pharmaceutical composition may be applied internally or externally.
  • the pharmaceutical composition is used in the form of an internal preparation, intravenous injection, subcutaneous injection, intradermal injection, injection such as intramuscular injection and / or intraperitoneal injection, transmucosal application, transdermal application and the like.
  • the dosage form of the pharmaceutical composition can be appropriately set depending on the form of application, for example, solid preparations such as tablets, granules, capsules, powders, powders, etc., liquid preparations such as solutions, suspensions, etc.
  • Semisolid agents such as ointments or gels are included.
  • one or more pharmaceutically acceptable additives can be freely selected and contained.
  • excipients for example, excipients, binders, disintegrants, surfactants, preservatives, coloring agents, flavoring agents, fragrances, stabilizers, preservatives
  • DDS drug delivery system
  • HMB 3-hydroxy-3-methylbutyrate
  • the muscle mass increasing agent of the present embodiment may be orally administered by a powder, capsule, tablet, granule, solution, syrup or the like.
  • the dose and administration form of the muscle mass increasing agent of the present embodiment may be appropriately selected depending on the subject, the pathological condition, the progress of the condition, and other conditions.
  • Euglena when Euglena is selected as an Euglena-derived substance and orally administered for the purpose of obtaining a muscle mass increasing effect for human (adult), Euglena is generally 1 to 5000 mg by dry weight per day, preferably It is recommended to administer continuously once or twice a day (morning and evening), and at a rate of 5 times a week or more, so as to be about 100 to 3000 mg, more preferably about 500 to 2000 mg.
  • Euglena powder was used as an Euglena-derived substance, and the influence of continuous intake of Euglena powder on muscle mass and muscle-related genes was examined.
  • Example 1 As a muscle mass increasing agent, Euglena gracilis powder (Euglena algal, manufactured by Euglena Co., Ltd.) which is a substance derived from Euglena was used.
  • Study 1 aims to clarify the effect of euglena intake on muscle mass in mice that are not disease models.
  • the quadriceps femoris muscle and liver after intake for 4 weeks were used as samples.
  • the control group was fed with CE-2 (normal diet), and the Euglena group was fed with CE-2 (Euglena diet) containing 2% Euglena (Table 4).
  • CE-2 normal diet
  • CE-2 Euglena diet
  • Table 4 After a 4-week feeding period, quadriceps muscles and liver were collected from the mice and organ weights were measured.
  • the quadriceps muscles are involved in the extension of the knee joint and the flexion of the hip joint, specifically, the action of stretching and standing the knee, the walking action, and the running action.
  • Euglena as an euglena-derived substance has an action of increasing the muscle mass of the quadriceps femoris muscle, and can improve movements such as standing up, walking, running and the like, thereby preventing and / or locomotive syndrome and sarcopenia. It has been found that it can be used to improve.
  • mice Male, 12 weeks old, 32 animals are divided into groups so that the average body weight is equal, and the normal rearing group (-), tail suspension group (C), tail suspension + Euglena low concentration group (EL) Tail suspension + Euglena high concentration group (EH).
  • Water is used for the normal rearing group and tail suspension group, and the euglena is 40 mg / mouse or 80 mg / mouse for tail suspension + low Euglena concentration (EL) and tail suspension + high euglena concentration (EH), respectively. It was orally administered once a day.
  • the tail suspension test was started 7 days after the start of administration, and 10 days after the start of the tail suspension test, it was sacrificed under isoflurane anesthesia.
  • RNA was extracted by abdominal aortic blood collection, and each muscle of liver, soleus muscle, plantar muscle, gastrocnemius muscle, and quadriceps femoris muscle was removed and weighed. After extracting RNA from the plantar and quadriceps muscles with Tri reagent, cDNA is synthesized, and each gene (Cbl-b (SEQ ID No. 1), Atrogin-1 (SEQ ID No. 2), MuRF1 (SEQ ID No.) is synthesized by real-time PCR. 3) The expression levels of 4E-BP1 (SEQ ID NO: 4) and RPS6KB1 (SEQ ID NO: 5) were measured. ⁇ -actin was used as an internal standard.
  • FIGS. The results of muscle weight measurements are shown in FIGS. As shown in FIG. 6, a decrease in muscle weight was observed in the tail suspension compared to the normal rearing group, and it was confirmed that the tail suspension induced disuse muscle atrophy. As shown in FIG. 7, in the plantar and quadriceps muscles, Euglena consumption inhibited the decrease in muscle weight. Euglena contains many components, which may have contributed to the inhibition of muscle protein degradation or the promotion of synthesis.
  • Euglena can be used to suppress the decrease in muscle weight of the plantar and quadriceps muscles.
  • Euglena can be used as a therapeutic agent or prophylactic agent for locomotive syndrome or sarcopenia, because reduction of muscle mass of plantar muscles and quadriceps leads to development of locomotive syndrome and sarcopenia.
  • the plantar muscles and quadriceps muscles are muscles that are strongly associated with motor function, it is possible to use Euglena as a motor function maintenance agent or a motor function improvement agent.
  • Euglena was selected from the ubiquitin ligase genes (Cbl-b (SEQ ID NO: 1), Atrogin-1 (SEQ ID NO: 2), MuRF1 (SEQ ID NO: 3)) involved in the degradation of muscle proteins. It has been found that it can be used as a muscle atrophy-related gene expression inhibitor for suppressing the expression of In addition, it was found that Euglena can be used as a muscle protein degradation inhibitor that inhibits the degradation of muscle protein and a muscle atrophy preventive agent that prevents muscle atrophy.
  • Cbl-b SEQ ID NO: 1
  • Atrogin-1 SEQ ID NO: 2
  • MuRF1 SEQ ID NO: 3
  • Euglena can be used as a muscle synthesis related gene expression promoter that promotes the expression of muscle synthesis related genes. Moreover, it turned out that it is possible to use Euglena as a muscle synthesis promoter which promotes muscle synthesis.

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Abstract

The present invention provides a food composition for increasing muscle mass, muscle mass increasing agent, therapeutic agent for locomotive syndrome, therapeutic agent for sarcopenia, exercise function maintenance agent, muscle atrophy-related gene expression inhibitor, muscle protein degradation inhibitor, muscle atrophy prevention agent, muscle synthesis-related gene expression promoter, and muscle synthesis promoter. A food composition for increasing muscle mass, muscle mass increasing agent, therapeutic agent for locomotive syndrome, therapeutic agent for sarcopenia, exercise function maintenance agent, muscle atrophy-related gene expression inhibitor, muscle protein degradation inhibitor, muscle atrophy prevention agent, muscle synthesis-related gene expression promoter, and muscle synthesis promoter, characterized in containing a Euglena-derived substance as an active ingredient.

Description

筋量増加用食品組成物、筋量増加剤、ロコモティブシンドローム治療剤、サルコペニア治療剤、運動機能維持剤、筋萎縮関連遺伝子発現抑制剤、筋タンパク質分解抑制剤、筋萎縮予防剤、筋合成関連遺伝子発現促進剤及び筋合成促進剤Food composition for increasing muscle mass, muscle mass increasing agent, locomotive syndrome therapeutic agent, sarcopenia therapeutic agent, motor function maintenance agent, muscle atrophy related gene expression inhibitor, muscle protein degradation inhibitor, muscle atrophy preventive agent, muscle synthesis related gene Expression promoter and muscle synthesis promoter

 本発明は、筋量増加用食品組成物、筋量増加剤、ロコモティブシンドローム治療剤、サルコペニア治療剤及び運動機能維持剤、筋萎縮関連遺伝子発現抑制剤、筋タンパク質分解抑制剤、筋萎縮予防剤、筋合成関連遺伝子発現促進剤及び筋合成促進剤に関する。 The present invention provides a food composition for increasing muscle mass, an agent for increasing muscle mass, an agent for treating locomotive syndrome, an agent for treating sarcopenia and a maintenance agent for motor function, an agent for suppressing muscle atrophy-related gene expression, an agent for suppressing muscle protein degradation, an agent for preventing muscle atrophy The present invention relates to a muscle synthesis related gene expression promoter and a muscle synthesis promoter.

 近年、加齢や運動不足に伴って生じる筋力の低下、筋肉量の減少が関連して生ずる症状である「サルコペニア」や「ロコモティブシンドローム」が大きな注目を集めている。
 サルコペニアとは、European Working Group on Sarcopenia in Older People(EWGSOP)によれば、筋肉量の低下に加え、筋肉の質の低下、すなわち筋力の低下あるいは身体的機能の低下がみられるものであると定義されている(非特許文献1)。
 ロコモティブシンドロームとは、日本整形外科学会によれば、骨、関節、軟骨、椎間板、筋肉といった運動器のいずれか、あるいは複数に障害が起こり、「立つ」、「歩く」といった機能が低下している状態であると定義されている(非特許文献2)。
In recent years, “sarcopenia” and “locomotive syndrome”, which are symptoms resulting from a decrease in muscle strength and a decrease in muscle mass caused by aging and lack of exercise, have attracted much attention.
Sarcopenia, according to the European Working Group on Sarcopenia in Older People (EWGSOP), is defined as a decrease in muscle quality, that is, a decrease in muscle quality, that is, a decrease in muscle strength or a decrease in physical function. (Non-Patent Document 1).
According to the Japanese Orthopedic Association, locomotive syndrome is a disorder in one or more of bone, joints, cartilage, intervertebral disc, muscles such as intervertebral disc, muscles, and functions such as "stand up" or "walk" are reduced. It is defined that it is a state (nonpatent literature 2).

 サルコペニアは加齢、疾病、運動不足、栄養不良などにより筋肉量が減少することに伴う筋力低下、身体的機能の低下を指し、ロコモティブシンドロームは、加齢や運動不足などにより筋肉、骨等の運動器の部位に支障をきたし、日常生活が困難になった状態を指すものである。つまり、ロコモティブシンドロームが運動器全般の症状を含む概念であるのに対し、サルコペニアはその運動器の中でも筋肉量、筋力、身体的機能に着目した症状であるといえる。 Sarcopenia refers to muscle weakness associated with loss of muscle mass due to aging, illness, lack of exercise, malnutrition, etc., and decrease in physical function, and locomotive syndrome refers to exercise of muscles, bones, etc. due to aging or lack of exercise It points to the state where troubles occurred in the part of the vessel and daily life became difficult. In other words, while locomotive syndrome is a concept that includes general motor symptoms, sarcopenia can be said to be a symptom that focuses on muscle mass, muscle strength, and physical function among the motor organs.

 ロコモティブシンドロームやサルコペニアを予防、改善する方法の一つとして、筋肉の増強がある。筋肉を増強するための筋肉増強剤として、ステロイドホルモンが知られているが、循環器疾患など副作用が生じることが報告されており、天然物由来の副作用のない素材の探索が行われている。例えば、タラ目に属する魚の魚肉タンパク質(特許文献1)や、リンゴ果汁圧搾残滓およびローズマリー抽出物(特許文献2)に筋肉量を増加させる作用があることが報告されている。 One of the ways to prevent and improve locomotive syndrome and sarcopenia is muscle strengthening. Although a steroid hormone is known as a muscle enhancing agent for enhancing muscles, side effects such as cardiovascular disease are reported to occur, and a search for materials having no side effects derived from natural products is being conducted. For example, it has been reported that the fish meat protein of a fish belonging to the order Coleoptera (Patent Document 1), and apple juice squeezed residue and rosemary extract (Patent Document 2) have an effect of increasing muscle mass.

 一方で、食糧、飼料、燃料等としての利用が有望視されている生物資源として、ユーグレナ(属名:Euglena、和名:ミドリムシ)が注目されている。
 ユーグレナは、ビタミン,ミネラル,アミノ酸,不飽和脂肪酸など、人間が生きていくために必要な栄養素の大半に該当する59種類もの栄養素を備え、多種類の栄養素をバランスよく摂取するためのサプリメントとしての利用や、必要な栄養素を摂取できない貧困地域での食糧供給源としての利用の可能性が提案されている。
On the other hand, Euglena (genus: Euglena, Japanese name: Midorimusi) is attracting attention as a biological resource which is considered to be promising for food, feed, fuel and the like.
Euglena is a supplement for the well-balanced intake of many types of nutrients, with 59 types of nutrients corresponding to most of the nutrients necessary for human beings to live including vitamins, minerals, amino acids, unsaturated fatty acids, etc. It has been suggested that it could be used as a food source in poor areas where it can not consume necessary nutrients.

 ユーグレナは、食物連鎖の第一次生産者に位置し、捕食者により捕食されることや、光、温度条件、撹拌速度などの培養条件が他の微生物に比べて難しいなどの理由から、大量培養が難しいとされてきたが、近年、本発明者らの鋭意研究によって、大量培養技術が確立され、ユーグレナ及びユーグレナから抽出されるパラミロン等、ユーグレナ由来物質の大量供給の途が開かれた。
 ユーグレナは、鞭毛運動をする動物的性質をもちながら、同時に植物として葉緑体を持ち光合成を行うユニークな生物であり、ユーグレナ自体やユーグレナ由来の物質に、多くの機能性があることが期待されている。
 そのため、大量供給可能となったユーグレナ及びユーグレナ由来物質の利用法の開発が望まれている。
Euglena is a primary producer of the food chain, and it is a large-scale culture because it is difficult to prey on predators and culture conditions such as light, temperature conditions, and agitation speed are higher than other microorganisms. In recent years, our extensive research has established a large-scale culture technique and opened the way to large-scale supply of Euglena-derived substances such as Euglena and Paramylon extracted from Euglena.
Euglena is a unique organism that has the property of carrying out flagellum movement and at the same time carries out photosynthesis with chloroplast as a plant, and it is expected that Euglena itself and substances derived from Euglena have many functions. ing.
Therefore, it is desirable to develop a method of using Euglena and Euglena-derived substances that can be supplied in large quantities.

特許第6109909号公報Patent No. 6109909 gazette 特許第5884112号公報Patent No. 5884112 gazette

神崎恒一、「サルコペニアの定義と診断方法」、日本医事新報、No.4677、pp.22-26(2013)Kozaki Koichi, "Defining and diagnostic methods of sarcopenia", Japanese Medical Journal, No. 4677, pp. 22-26 (2013) 公益社団法人日本整形外科学会、「ロコモティブシンドローム」、ロコモパンフレット2015年度版Japan Orthopedic Association, "Locomotive Syndrome", Locomo Brochure 2015 Edition

 本発明は、上記の課題に鑑みてなされたものであり、本発明の目的は、日常的に継続して摂取することができ、効果的に筋肉量を増加させ、ロコモティブシンドロームやサルコペニアの治療、予防、改善をすることが可能な筋量増加用食品組成物、筋量増加剤、ロコモティブシンドローム治療剤、サルコペニア治療剤、運動機能維持剤、筋萎縮関連遺伝子発現抑制剤、筋タンパク質分解抑制剤、筋萎縮予防剤、筋合成関連遺伝子発現促進剤及び筋合成促進剤を提供することにある。 The present invention has been made in view of the above problems, and the object of the present invention is to be able to be taken continuously continuously, to effectively increase muscle mass, to treat locomotive syndrome and sarcopenia, Food composition for increasing muscle mass that can be prevented and improved, muscle mass increasing agent, locomotive syndrome therapeutic agent, sarcopenia therapeutic agent, motor function maintenance agent, muscle atrophy related gene expression inhibitor, muscle protein degradation inhibitor, An object of the present invention is to provide a muscle atrophy preventive agent, a muscle synthesis related gene expression promoter and a muscle synthesis promoter.

 本発明者らは、上記課題を解決すべく鋭意研究した結果、ユーグレナに筋肉量を増加させる作用、筋萎縮を抑制する作用、筋タンパク質の分解を抑制する作用、筋合成を促進する作用があることを見出し、本発明を完成するに至った。 As a result of earnest studies to solve the above problems, the present inventors have an effect of increasing muscle mass in Euglena, an action of suppressing muscle atrophy, an action of suppressing degradation of muscle proteins, and an action of promoting muscle synthesis. The present invention has been completed.

 従って、前記課題は、本発明によれば、ユーグレナ由来物質を有効成分として含有することを特徴とする筋量増加用食品組成物により解決される。
 このとき、前記ユーグレナ由来物質がユーグレナであるとよい。
 このとき、大腿四頭筋の量を増加させるために用いられる筋量増加用食品組成物であるとよい。
 また、前記課題は、本発明によれば、ユーグレナ由来物質を有効成分として含有することを特徴とする筋量増加剤により解決される。
Therefore, according to the present invention, the above-mentioned subject is solved by the food composition for increasing muscle mass characterized by containing an Euglena-derived substance as an active ingredient.
At this time, the Euglena-derived substance is preferably Euglena.
At this time, it may be a muscle mass increasing food composition used to increase the amount of quadriceps.
Further, according to the present invention, the above-mentioned problems are solved by a muscle mass increasing agent characterized by containing an Euglena-derived substance as an active ingredient.

 また、前記課題は、本発明によれば、ユーグレナ由来物質を有効成分として含有し、サルコペニア予防用、サルコペニア改善用、ロコモティブシンドローム予防用又はロコモティブシンドローム改善用の食品組成物により解決される。
 また、前記課題は、本発明によれば、ユーグレナ由来物質を有効成分として含有することを特徴とするロコモティブシンドローム予防剤、ロコモティブシンドローム治療剤、サルコペニア予防剤、サルコペニア治療剤、運動機能維持剤及び運動機能改善剤により解決される。
Further, according to the present invention, the above problems are solved by a food composition containing an Euglena-derived substance as an active ingredient, for sarcopenia prevention, sarcopenia improvement, locomotive syndrome prevention or locomotive syndrome improvement.
Further, according to the present invention, according to the present invention, a locomotive syndrome preventive agent, a locomotive syndrome therapeutic agent, a sarcopenia preventive agent, a sarcopenia therapeutic agent, a motor function maintenance agent, exercise comprising the euglena-derived substance as an active ingredient. It is solved by the function improving agent.

 また、前記課題は、本発明によれば、ユーグレナ由来物質を有効成分として含有し、筋萎縮関連遺伝子発現抑制用、筋タンパク質分解抑制用、筋萎縮予防用、筋合成関連遺伝子発現促進用又は筋合成促進用の食品組成物により解決される。
 また、前記課題は、本発明によれば、ユーグレナ由来物質を有効成分として含有することを特徴とする筋萎縮関連遺伝子発現抑制剤、筋タンパク質分解抑制剤、筋萎縮予防剤、筋合成関連遺伝子発現促進剤及び筋合成促進剤により解決される。
Further, according to the present invention, the subject contains an Euglena-derived substance as an active ingredient, and is for suppressing muscle atrophy-related gene expression, for suppressing muscle protein degradation, for preventing muscle atrophy, for promoting muscle synthesis-related gene expression or It is solved by a food composition for promoting synthesis.
Further, according to the present invention, according to the present invention, a muscle atrophy-related gene expression inhibitor characterized by containing an Euglena-derived substance as an active ingredient, a muscle protein degradation inhibitor, a muscle atrophy preventive agent, a muscle synthesis related gene expression It is solved by the promoter and the muscle synthesis promoter.

 本発明によれば、新規な筋量増加用食品組成物、筋量増加剤、ロコモティブシンドローム治療剤、サルコペニア治療剤、運動機能維持剤、筋萎縮関連遺伝子発現抑制剤、筋タンパク質分解抑制剤、筋萎縮予防剤、筋合成関連遺伝子発現促進剤及び筋合成促進剤を提供することができる。
 本発明の筋量増加用食品組成物、筋量増加剤、ロコモティブシンドローム治療剤、サルコペニア治療剤、運動機能維持剤、筋萎縮関連遺伝子発現抑制剤、筋タンパク質分解抑制剤、筋萎縮予防剤、筋合成関連遺伝子発現促進剤及び筋合成促進剤は、これまでに副作用の報告がなく、食品衛生法に合致する水準の安全性を備えたユーグレナを有効成分としているため、長期間の継続投与および継続摂取が可能である。
According to the present invention, a novel food composition for increasing muscle mass, muscle mass increasing agent, locomotive syndrome therapeutic agent, sarcopenia therapeutic agent, motor function maintenance agent, muscle atrophy related gene expression inhibitor, muscle protein degradation inhibitor, muscle The agent for preventing atrophy, muscle synthesis related gene expression promoter and muscle synthesis promoter can be provided.
The food composition for increasing muscle mass of the present invention, muscle mass increasing agent, locomotive syndrome therapeutic agent, sarcopenia therapeutic agent, motor function maintenance agent, muscle atrophy related gene expression inhibitor, muscle protein degradation inhibitor, muscle atrophy preventive agent, muscle The synthetic related gene expression promoter and muscle synthesis promoter have no reports of adverse effects and have Euglena as the active ingredient with a level of safety that conforms to the Food Sanitation Law. It can be taken.

試験1において、各試験食を継続して4週間摂食させた時の体重変化を示すグラフである。In Experiment 1, it is a graph which shows a body weight change when each test meal is continuously fed for 4 weeks. 試験1における試験食の摂食量を示すグラフである。6 is a graph showing the amount of food consumed by the test food in Test 1. 試験1終了時における肝臓重量を示すグラフである。It is a graph which shows the liver weight at the end of test 1. 試験1終了時における大腿四頭筋重量を示すグラフである(Tukey‘s test、*p<0.05 vs.コントロール群)。It is a graph which shows quadriceps femoris muscle weight at the end of test 1 (Tukey's test, * p <0.05 vs. control group). 試験2試験終了時における体重及び肝臓重量を示すグラフである(Data are means ±SE,n=6,Tukey’s test,**p<0.01 vs.通常飼育群(-))。Test 2 A graph showing body weight and liver weight at the end of the test (Data are means ± SE, n = 6, Tukey's test, ** p <0.01 vs. ordinary rearing group (−)). 試験2終了後の各筋肉重量を示すグラフである(Data are means ±SE,n=6,Tukey’stest,**p<0.01,***p<0.001 vs.通常飼育群(-))。It is a graph which shows each muscle weight after the completion of examination 2 (Data are means ± SE, n = 6, Tukey's test, ** p <0.01, *** p <0.001 vs. ordinary rearing group ( -)). 試験2終了後の通常飼育群(-)に対する各筋肉重量の割合を示すグラフである(Data are means ±SE,n=6,Dunnett multiple test,†p<0.05,††p<0.01 vs.尾懸垂群(C))。It is a graph which shows the ratio of each muscle weight to the normal breeding group (-) after the completion of test 2 (Data are means ± SE, n = 6, Dunnett multiple test, p p <0.05, † p <0. 01 vs. tail suspension group (C)). 試験2終了後の各筋肉における筋萎縮関連遺伝子の発現量を示すグラフである(Data are means ±SE,n=6,Tukey’s test,*p<0.05,**p<0.01,***p<0.001 vs.尾懸垂群(C))。It is a graph which shows the expression level of the muscle atrophy related gene in each muscle after the completion of test 2 (Data are means ± SE, n = 6, Tukey's test, * p <0.05, ** p <0.01 , *** p <0.001 vs. tail suspension group (C)). 筋タンパク質の分解(筋萎縮)のメカニズムを示す説明図である。It is explanatory drawing which shows the mechanism of decomposition | disassembly of muscle protein (muscle atrophy). 試験2終了後の各筋肉における筋合成関連遺伝子の発現量を示すグラフである(Data are means ±SE,n=6,Tukey’s test,#p<0.05 vs.TS(Cを除いた3群比較:Dunnett multiple test,†p<0.05,††p<0.01 vs.TS))。It is a graph which shows the expression level of the muscle synthesis related gene in each muscle after the end of test 2 (Data are means ± SE, n = 6, Tukey's test, # p <0.05 vs. TS (C is excluded) Three group comparison: Dunnett multiple test, p p <0.05, † p <0.01 vs. TS)). 筋タンパク質の合成(筋合成)のメカニズムを示す説明図である。It is an explanatory view showing a mechanism of muscle protein synthesis (muscle synthesis).

 以下、本発明の実施形態について、図1乃至11を参照しながら説明する。
 本実施形態は、ユーグレナ由来物質を有効成分として含有する筋量増加用食品組成物、筋量増加剤、ロコモティブシンドローム治療剤、サルコペニア治療剤、運動機能維持剤、筋萎縮関連遺伝子発現抑制剤、筋タンパク質分解抑制剤、筋萎縮予防剤、筋合成関連遺伝子発現促進剤及び筋合成促進剤に関するものである。
Hereinafter, embodiments of the present invention will be described with reference to FIGS.
In this embodiment, a muscle mass increasing food composition containing an Euglena-derived substance as an active ingredient, a muscle mass increasing agent, a locomotive syndrome therapeutic agent, a sarcopenia therapeutic agent, a motor function maintenance agent, a muscle atrophy-related gene expression inhibitor, a muscle The present invention relates to a protein degradation inhibitor, a muscle atrophy preventive agent, a muscle synthesis related gene expression promoter and a muscle synthesis promoter.

<ロコモティブシンドローム>
 ロコモティブシンドローム(locomotive syndrome)とは、日本整形外科学会によれば、骨、関節、軟骨、椎間板、筋肉といった運動器のいずれか、あるいは複数に障害が起こり、「立つ」、「歩く」といった機能が低下している状態であると定義されている(非特許文献2)。
 ロコモティブシンドロームは、ロコモと略されたり、ロコモティブ症候群、運動器症候群などと呼ばれることもある。
 以下、ロコモティブシンドロームの診断基準について説明する(吉村典子、「ロコモティブシンドロームの臨床診断値と有病率」、日本老年医学会雑誌52巻4号、350-353頁(2015))。
<Locomotive syndrome>
According to the Japanese Orthopedic Association, locomotive syndrome (locomotive syndrome) is characterized by the ability to “stand up” or “walk” due to injury to one or more of bone, joint, cartilage, intervertebral disc, muscle, or muscle It is defined as being in a state of decline (Non-Patent Document 2).
Locomotive syndrome may be abbreviated as Locomo, or may be called locomotive syndrome, motor organ syndrome or the like.
Hereinafter, diagnostic criteria for locomotive syndrome will be described (Noriko Yoshimura, "Clinical diagnostic value and prevalence of locomotive syndrome", Japanese Geriatrics Journal 52, 4 350-353 (2015)).

 (ロコモ度テスト)
 ロコモティブシンドロームの診断基準の一つとなるロコモ度テストは、(1)立ち上がりテスト、(2)2ステップテスト、(3)ロコモ25からなる。
 (1)立ち上がりテストは、10cm、20cm、30cm、40cmの4つの高さの台を準備して、片脚または両脚で立ち上がれるかどうかで脚力を測るテストである。
 (2)2ステップテストは、できるかぎり大股で2歩歩き、2歩分の歩幅を測定し、身長で除して2ステップ値を算出する。2ステップ値により,下肢の筋力、バランス能力、柔軟性などを含めた歩行能力を評価する。
 (3)ロコモ25は、過去1カ月の間に体の痛みや日常生活の困難がなかったかどうかについて25項目の問診票で評価する。
(Locomo test)
The locomotion test, which is one of diagnostic criteria for locomotive syndrome, comprises (1) rising test, (2) two-step test, and (3) locomo 25.
(1) Stand-up test is a test of measuring the leg strength based on whether one leg or both legs can stand up by preparing a stand of four heights of 10 cm, 20 cm, 30 cm and 40 cm.
(2) In the two-step test, walk two steps with as much as possible, measure the stride for two steps, and divide by the height to calculate a two-step value. Evaluate walking ability including leg strength, balance ability, flexibility etc. of lower limbs by 2-step value.
(3) Locomo 25 evaluates with a 25-item questionnaire about whether or not there was body pain or difficulty in daily life in the past one month.

 ロコモ度テストによる判断は、ロコモ度1とロコモ度2の2つからなる。
 ロコモ度1の臨床判断値は以下の通りである。
1)立ち上がりテスト:どちらか一方の片脚で40cmの高さから立ち上がれない
2)2ステップテスト:2ステップ値が1.3未満
3)ロコモ25:ロコモ25の結果が7点以上
 上記1)~3)のいずれかひとつでもあてはまる場合は「ロコモ度1」と判定され、移動機能の低下が始まっている状態と判断される。
The judgment based on the locomotion test consists of 2 locomotions 1 and 2.
The clinical judgment value of locomotion degree 1 is as follows.
1) Rise test: Can not get up from a height of 40 cm with either one leg 2) 2 step test: 2 step value less than 1.3 3) Locomo 25: Locomo 25 result is 7 points or more Above 1)- If any one of the above 3) applies, it is determined as "Locomor 1", and it is determined that the mobile function has started to deteriorate.

 ロコモ度2の臨床判断値は以下の通りである。
1)立ち上がりテスト:両脚で20cmの高さから立ち上がれない
2)2ステップテスト:2ステップ値が1.1未満
3)ロコモ25:ロコモ25の結果が16点以上
 上記1)~3)のいずれかひとつでもあてはまる場合は「ロコモ度2」と判定され、移動機能の低下が進行している状態と判断される。
The clinical judgment value of locomotion degree 2 is as follows.
1) Rise test: Can not stand from height of 20 cm with both legs 2) 2 step test: 2 step value less than 1.1 3) Locomo 25: Locomo 25 result is 16 points or more Any of the above 1) to 3) If any one of the cases is true, it is judged as "Locomor 2", and it is judged that the deterioration of the mobile function is progressing.

<サルコペニア>
 サルコペニア(sarcopenia)とは、加齢に伴い骨格筋が萎縮し、骨格筋量及び骨格筋力の低下または身体機能の低下を伴うことを特徴とする症候群である。
 サルコペニアは、国や地域などによって異なる多く診断基準が提唱されているが、筋肉量の低下を必須項目とし、筋力または身体能力の低下のいずれかに該当した場合にサルコペニアと診断される(葛谷雅文、「サルコペニアの診断・病態・治療」日本老年医学会雑誌52巻4号、343-349頁(2015))。
 以下、サルコペニアの診断基準について説明する(公益財団法人長寿科学振興財団、健康長寿ネット、サルコペニアの診断、URL:https://www.tyojyu.or.jp/net/byouki/sarcopenia/shindan.html)。
<Sarcopenia>
Sarcopenia is a syndrome characterized by atrophy of skeletal muscle with age, accompanied by loss of skeletal muscle mass and muscle strength or loss of physical function.
For sarcopenia, many diagnostic criteria that differ depending on the country, region, etc. have been proposed, but it is considered as a sarcopenia when muscle loss is an essential item, and it falls under either muscle strength or physical ability "Sarcopenia's diagnosis, pathophysiology and treatment" Journal of Japan Geriatrics Journal Vol. 52, No. 4, 343-349 (2015)).
The following is a description of the diagnostic criteria for sarcopenia (The Foundation for Longevity Science Promotion, Health Longevity Net, Diagnosis of sarcopenia, URL: https://www.tyojyu.or.jp/net/byouki/sarcopenia/shindan.html) .

(EWGSOPによる診断基準)
 EWGSOPによれば、四肢骨格筋量の低下があることに加えて身体機能(歩行速度)の低下または、筋力(握力)の低下がある場合にサルコペニアと診断される。
 具体的には、四肢骨格筋量(ALM)は四肢の筋肉量を身長(m)の2乗で割った、二重エネルギーX線吸収測定法(DXA)の値(kg/m)が男性では7.23kg/m以下の場合、女性では5.67kg/m以下であることが必須の条件であり、さらに10mの歩行速度が0.8m/秒未満の場合、あるいは握力が男性では30kg未満、女性では20kg未満の場合にサルコペニアと診断される。
(Diagnostic criteria by EWGSOP)
According to the EWGSOP, if there is a decrease in physical function (walking speed) or a decrease in muscle strength (grip strength) in addition to a decrease in limb skeletal muscle mass, a diagnosis of sarcopenia is made.
Specifically, the limb skeletal muscle mass (ALM) is the muscle mass of the limb divided by the height (m) squared, and the dual energy X-ray absorptiometry (DXA) value (kg / m 2 ) is male in the case of 7.23kg / m 2 or less, in women is an essential condition to be less 5.67kg / m 2, further when the walking speed of 10m is less than 0.8 m / sec, or grip strength in men Sarcopenia is diagnosed if it is less than 30 kg and less than 20 kg in women.

 診断の手順は、最初に歩行速度を測定して0.8m/秒以下の場合は四肢骨格筋量を測定して、診断基準の男性で7.23kg/m以下、女性で5.67kg/m以下の場合はサルコペニア、高値の場合はサルコペニアではないと診断される。
 歩行速度が0.8m/秒より早かった場合には、握力を測定し、男性で30kg、女性で20kgよりも高値の場合はサルコペニアではないと診断され、高値の場合は四肢骨格筋量を測定し、診断基準値以下の場合はサルコペニア、診断基準値以上の場合はサルコペニアではないと診断される。
The procedure of diagnosis is to measure the walking speed first and measure limb skeletal muscle mass if it is less than 0.8 m / s, and it is 7.23 kg / m 2 or less for men and 5.67 kg / m for women. If it is m 2 or less, sarcopenia is diagnosed. If it is high, it is not diagnosed as sarcopenia.
If walking speed is faster than 0.8 m / sec, grip strength is measured and if it is higher than 30 kg for males and 20 kg for females it is not diagnosed as sarcopenia, if it is higher, limb skeletal muscle mass is measured Sarcopenia if below the diagnostic threshold, not sarcopenia if above the diagnostic threshold.

(アジア人の診断基準)
 EWGSOPの診断基準は、ヨーロッパのワーキンググループによって作られた基準であり、ヨーロッパ人とアジア人では体格や身体機能に違いがあるため、アジアのサルコペニアについてのワーキンググループ、ASIAN working Group FOR SARCOPENIA(AWGS)によってアジア人向けの診断基準が作られている。AWGSによる診断基準では、EWGSOPの診断基準と握力、筋肉量の基準が異なっている。
(Diagnostic criteria for Asians)
The diagnostic criteria of the EWGSOP are the criteria developed by the working group in Europe, and because there is a difference in physique and physical function between Europeans and Asians, the working group for sarcopenia in Asia, ASIAN working Group FOR SARCPOPENIA (AWGS) Has established diagnostic criteria for Asians. The AWGS diagnostic criteria differ from the EWGSOP diagnostic criteria and the criteria for grip strength and muscle mass.

 サルコペニアと診断される握力の基準は、男性で26kg未満、女性で18kg未満であり、筋肉量の基準は、男性で7kg/m以下(DXA値,BIA値)、女性で5.4kg/m以下(DXA値),5.7kg/m以下(BIA値)とされている。 The standard of grip strength diagnosed with sarcopenia is less than 26 kg for men and less than 18 kg for women, and the criteria for muscle mass are 7 kg / m 2 or less for men (DXA value, BIA value), 5.4 kg for women 2 or less (DXA value), 5.7 kg / m 2 or less (BIA value).

(日本人の診断基準)
 EWGSOPの診断基準は欧米人の高齢者を基準としており、日本人と欧米人では、高齢者であっても体格や生活習慣の違いがあるため、日本人の高齢者に合ったサルコペニアの簡易基準案が国立長寿医療研究センター・老化に関する長期縦断疫学研究(NILS-LSA)によって作成されている。
(Japanese diagnostic criteria)
The diagnostic criteria in the EWGSOP are based on the elderly of Western Americans, and Japanese and European Americans have differences in physique and lifestyle even with the elderly, so the simplified criteria of sarcopenia suitable for the elderly of Japanese A draft has been prepared by the National Longevity Medical Research Center Long-Term Longitudinal Epidemiological Study on Aging (NILS-LSA).

 NILS-LSAによる診断基準では、65歳以上の高齢者で、歩行速度が1m/秒未満、もしくは握力が男性で25kg未満、女性で20kg未満である場合で、さらにBMI値(ボディマス指数、身長の二乗に対する体重の比で体格を表す指数)が18.5未満、もしくは下腿囲が30cm未満の場合にサルコペニアと診断される。
 歩行速度、握力が基準値以上であった場合は正常。歩行速度、握力が基準値以下でもBMI、下腿囲が基準値以上であれば脆弱高齢者であるがサルコペニアではないと診断される。
According to the NILS-LSA diagnostic criteria, BMI values (body mass index, height) for elderly people over the age of 65 with walking speeds less than 1 m / s, or grips less than 25 kg for men and less than 20 kg for women. A diagnosis of sarcopenia is made if the ratio of body weight to the square is an index representing body size) less than 18.5, or the lower leg circumference is less than 30 cm.
Normal when walking speed and grip strength are above standard values. Even if the walking speed and grip strength are below the standard value, BMI and lower leg circumference are above the standard value, it is diagnosed as a vulnerable elderly person but not sarcopenia.

<ユーグレナ>
 本実施形態において、「ユーグレナ」とは、動物学や植物学の分類でユーグレナ属(Euglena)に分類される微生物、その変種、その変異種のすべてを含む。
 ここで、ユーグレナ属(Euglena)の微生物とは、動物学では原生動物門(Protozoa)の鞭毛虫綱(Mastigophorea)、植物鞭毛虫亜綱(Phytomastigophorea)に属するミドリムシ目(Euglenida)のユーグレノイディナ亜目(Euglenoidina)に属する微生物である。一方、ユーグレナ属の微生物は、植物学ではミドリムシ植物門(Euglenophyta)のミドリムシ藻類綱(Euglenophyceae)に属するミドリムシ目(Euglenales)に属している。
<Euglena>
In the present embodiment, "Euglena" includes all microorganisms classified into Euglena (Euglena) according to the zoological and botanical classification, variants thereof, and variants thereof.
Here, the microorganisms of Euglena (Euglena) are, in zoology, the protozoan (Protozoa) dinoflagellate (Mastigophorea), and the Euglenoidina (Euglenida) belonging to the plant dinoflagellate (Phytomastigophorea) It is a microorganism belonging to the eye (Euglenoidina). On the other hand, microorganisms of the genus Euglena belong to the order Euglenales belonging to the family Euglenophyceae of the family Agromorpha (Euglenophyta) in botany.

 ユーグレナ属の微生物としては、具体的には、Euglena acus、Euglena anabaena、Euglena caudata、Euglena chadefaudii、Euglena deses、Euglena gracilis、Euglena granulata、Euglena intermedia、Euglena mutabilis、Euglena oxyuris、Euglena proxima、Euglena spirogyra、Euglena viridis、Euglena vermiformis、Euglena intermedia, Euglena pirideなどが挙げられる。
 ユーグレナ細胞としては、ユーグレナ・グラシリス(E. gracilis)、特に、ユーグレナ・グラシリス(E. gracilis)Z株、ユーグレナ・グラシリス(E. gracilis)NIES-49株などを用いることができるが、そのほか、ユーグレナ・グラシリス(E. gracilis)Z株の変異株SM-ZK株(葉緑体欠損株)や変種のvar. bacillaris、これらの種の葉緑体の変異株等の遺伝子変異株由来のβ-1,3-グルカナーゼ、Euglena intermedia, Euglena piride、及びその他のユーグレナ類、例えばAstaia longaであってもよい。
Specifically, Euglena acus, Euglena anabaena, Euglena caudata, Euglena chadefaudii, Euglena deses, Euglena gracilis, Euglena granulata, Euglena mutulalis, Euglena muturilis, Euglena oxiuris, Euglena smith, as microorganisms of the genus Euglena. , Euglena vermiformis, Euglena intermedia, Euglena piride and the like.
As Euglena cells, Euglena gracilis (E. gracilis), in particular, E. gracilis (E. gracilis) Z strain, E. gracilis (E. gracilis) NIES-49 strain, etc. can be used, but in addition, Euglena · Mutant strain SM-ZK of E. gracilis Z strain (chloroplast-deficient strain) or variant var. Bacillaris, β-1 derived from gene mutant strains such as chloroplast mutant strains of these species , 3-glucanase, Euglena intermedia, Euglena piride, and other Euglenas, such as Astaia longa.

 ユーグレナ属は、池や沼などの淡水中に広く分布しており、これらから分離して使用しても良く、また、既に単離されている任意のユーグレナ属を使用してもよい。
 ユーグレナ属は、その全ての変異株を包含する。また、これらの変異株の中には、遺伝的方法、たとえば組換え、形質導入、形質転換等により得られたものも含有される。
The genus Euglena is widely distributed in fresh water such as ponds and marshes, and may be used separately from them, or any Euglena isolated previously may be used.
The genus Euglena includes all its variants. Further, among these mutant strains, those obtained by genetic methods such as recombination, transduction, transformation and the like are also contained.

 ユーグレナ細胞の培養において、培養液としては、例えば、窒素源,リン源,ミネラルなどの栄養塩類を添加した培養液、例えば、改変Cramer-Myers培地((NHHPO 1.0g/L,KHPO 1.0g/L,MgSO・7HO 0.2g/l,CaCl・2HO 0.02g/l,Fe(SO・7HO 3mg/l,MnCl・4HO 1.8mg/l,CoSO・7HO 1.5mg/l,ZnSO・7HO 0.4mg/l,NaMoO・2HO 0.2mg/l,CuSO・5HO 0.02g/l,チアミン塩酸塩(ビタミンB1) 0.1mg/l,シアノコバラミン(ビタミンB12)、(pH3.5))を用いることができる。なお、(NHHPOは、(NHSOやNHaqに変換することも可能である。また、そのほか、ユーグレナ 生理と生化学(北岡正三郎編、(株)学会出版センター)の記載に基づき調製される公知のHutner培地,Koren-Hutner培地を用いてもよい。 In the culture of Euglena cells, as a culture solution, for example, a culture solution to which nutrients such as nitrogen source, phosphorus source, and minerals have been added, for example, modified Cramer-Myers medium ((NH 4 ) 2 HPO 4 1.0 g / L , KH 2 PO 4 1.0 g / L, MgSO 4 · 7 H 2 O 0.2 g / l, CaCl 2 · 2 H 2 O 0.02 g / l, Fe 2 (SO 2 ) 3 · 7 H 2 O 3 mg / l, MnCl 2 · 4 H 2 O 1.8 mg / l, CoSO 4 · 7 H 2 O 1.5 mg / l, ZnSO 4 · 7 H 2 O 0.4 mg / l, Na 2 MoO 4 · 2 H 2 O 0.2 mg / l, CuSO 4 · 5H 2 O 0.02g / l, thiamine hydrochloride (vitamin B1) 0.1mg / l, cyanocobalamin (vitamin B12), can be used (pH 3.5)). (NH 4 ) 2 HPO 4 can also be converted to (NH 4 ) 2 SO 4 or NH 3 aq. In addition, well-known Hutner's medium and Koren-Hutner's medium prepared based on the description of Euglena Physiology and Biochemistry (Kitaoka Shozaburo, ed., Inc. publication center) may be used.

 培養液のpHは好ましくは2以上、また、その上限は、好ましくは6以下、より好ましくは4.5以下である。pHを酸性側にすることにより、光合成微生物は他の微生物よりも優勢に生育することができるため、コンタミネーションを抑制できる。
 ユーグレナ細胞の培養は、太陽光を直接利用するオープンポンド方式、集光装置で集光した太陽光を光ファイバー等で送り、培養槽で照射させ光合成に利用する集光方式等により行っても良い。
 また、ユーグレナ細胞の培養は、例えば供給バッチ法を用いて行われ得るが、フラスコ培養や発酵槽を用いた培養、回分培養法、半回分培養法(流加培養法)、連続培養法(灌流培養法)等、いずれの液体培養法により行っても良い。
 ユーグレナ細胞の分離は、例えば培養液の遠心分離,濾過又は単純な沈降によって行われる。
The pH of the culture solution is preferably 2 or more, and the upper limit thereof is preferably 6 or less, more preferably 4.5 or less. By setting the pH to the acidic side, photosynthetic microorganisms can be more preferentially grown than other microorganisms, so that contamination can be suppressed.
The culture of Euglena cells may be performed by an open pond method using sunlight directly, a method of sending sunlight collected by a light collecting device by an optical fiber or the like and irradiating it in a culture tank and utilizing it for photosynthesis.
In addition, culture of Euglena cells may be performed using, for example, a fed-batch method, but culture using a flask culture or a fermenter, batch culture method, half-batch culture method (feed-batch culture method), continuous culture method (perfusion It may be carried out by any liquid culture method such as culture method).
The separation of Euglena cells is performed, for example, by centrifugation, filtration or simple sedimentation of the culture fluid.

(ユーグレナ藻体)
 本実施形態では、ユーグレナ藻体として、遠心分離,濾過又は沈降等によって分離したユーグレナ生細胞をそのまま用いることができる。ユーグレナ生細胞は、培養槽から収穫後そのままの状態で使用することもできるが、水若しくは生理食塩水で洗浄するのが好ましい。また、ユーグレナ藻体が水などの液体に分散した分散液の状態で用いてもよい。本実施形態において、ユーグレナ生細胞を凍結乾燥処理やスプレー乾燥処理して得たユーグレナの乾燥藻体をユーグレナ藻体として用いると好適である。
 更に、ユーグレナ生細胞を超音波照射処理や、ホモゲナイズ等の機械処理を行うことにより得た藻体の機械的処理物をユーグレナ藻体として用いてもよい。また、機械的処理物に乾燥処理を施した機械的処理物乾燥物をユーグレナ藻体として用いてもよい。
(Euglena algae)
In this embodiment, Euglena live cells separated by centrifugation, filtration, sedimentation or the like can be used as Euglena algal cells as they are. Euglena living cells can be used as they are after harvesting from a culture tank, but washing with water or saline is preferred. Also, it may be used in the form of a dispersion in which Euglena algal cells are dispersed in a liquid such as water. In the present embodiment, it is preferable to use the dried algal cells of Euglena obtained by freeze-drying treatment or spray-drying treatment of Euglena live cells as Euglena algal cells.
Furthermore, a mechanically treated product of algal cells obtained by performing ultrasonic irradiation treatment or mechanical treatment such as homogenization may be used as the euglena algal cells. Moreover, you may use the mechanical-process dried material which performed the dry process to the mechanical process as an Euglena algal body.

(ユーグレナ水性溶媒抽出物)
 本実施形態において、「ユーグレナ水性溶媒抽出物」とは、水性溶媒を用いてユーグレナから抽出される抽出物を意味し、特に、水性溶媒として水を用い、5℃~600℃で、数秒~数十時間抽出したユーグレナの水抽出物又は熱水抽出物を用いることが好ましい。
 抽出に使用する水は、必ずしも蒸留水や、純水、又は超純水である必要はなく、例えば、水道水や不純物を含むものであってもよいが、活性成分の抽出を妨げる成分を含まない水が好ましい。
(Euglena aqueous solvent extract)
In the present embodiment, “Euglena aqueous solvent extract” means an extract extracted from Euglena using an aqueous solvent, and in particular, using water as an aqueous solvent, several seconds to several seconds at 5 ° C. to 600 ° C. It is preferred to use a water extract or a hot water extract of Euglena extracted for 10 hours.
The water used for the extraction does not necessarily have to be distilled water, pure water, or ultrapure water, and may be, for example, tap water or an impurity, but contains a component that hinders the extraction of the active ingredient. No water is preferred.

 本実施形態において、「水抽出物」とは、0~50℃(0℃を除く。)の水による抽出物を意味する。
 ここで、「水」とは、0~50℃(0℃を除く。)の水を意味する。
 水の温度は、活性成分に影響を与えずに、活性成分を十分に抽出できる範囲内であれば特に限定されるものではないが、好ましくは1~40℃、より好ましくは5~35℃、特に好ましくは10~30℃である。
In the present embodiment, “water extract” means an extract with water of 0 to 50 ° C. (excluding 0 ° C.).
Here, “water” means water of 0 to 50 ° C. (excluding 0 ° C.).
The temperature of water is not particularly limited as long as the active ingredient can be sufficiently extracted without affecting the active ingredient, but preferably 1 to 40 ° C., more preferably 5 to 35 ° C. Particularly preferred is 10 to 30 ° C.

 本実施形態において、「熱水抽出物」とは、50℃よりも高い温度の水による抽出物を意味し、「温水抽出物」とも呼ぶことができる。
 ここで、「熱水」とは、50℃よりも高温の水を意味し、「熱湯」も含む概念であり、沸騰状態にある水も含まれる。また、液体状態の熱水に限定されることなく、気体状態及び超臨界状態の熱水も含まれる。
 熱水の温度は、活性成分に影響を与えずに、活性成分を十分に抽出できる範囲内であれば特に限定されるものではないが、好ましくは50℃より高く120℃以下、より好ましくは50℃より高く100℃以下である。
In the present embodiment, “hot water extract” means an extract with water at a temperature higher than 50 ° C., and can also be called “hot water extract”.
Here, “hot water” means water having a temperature higher than 50 ° C., and is a concept including “hot water”, including water in a boiling state. Further, it is not limited to hot water in a liquid state, and includes hot water in a gaseous state and in a supercritical state.
The temperature of the hot water is not particularly limited as long as the active ingredient can be sufficiently extracted without affecting the active ingredient, but is preferably higher than 50 ° C. and 120 ° C. or less, more preferably 50 It is higher than ° C and not higher than 100 ° C.

 抽出に使用する水のpHは、活性成分に影響を与えずに、活性成分を十分抽出できる範囲内であれば特に限定されるものではないが、好ましくはpH4~10、より好ましくはpH5~9、特に好ましくはpH6~8であるとよい。 The pH of water used for extraction is not particularly limited as long as the active ingredient can be extracted sufficiently without affecting the active ingredient, but the pH is preferably 4 to 10, more preferably 5 to 9 The pH is particularly preferably 6 to 8.

 なお、本実施形態では、水性溶媒として、水を単独で用いるが、活性成分に影響を与えずに、活性成分を十分抽出できるものであって、通常、抽出に用いることができる溶媒を1種または2種以上選択して用いてもよい。例えば、水、アルコール類、グリコール類などを挙げることができるが、これに限定されるものではない。アルコール類としては、エタノール、メタノール、n-プロパノール、イソプロパノール等が挙げられる。グリコール類としては、ブチレングリコール及びプロピレングリコール等が挙げられる。その他の水性溶媒としては、アセトン等が挙げられる。これらの溶媒は単独或いは水溶液として用いても良く、任意の2種または3種以上の混合溶媒として用いてもよい。 In the present embodiment, water is used alone as the aqueous solvent, but the active ingredient can be sufficiently extracted without affecting the active ingredient, and one solvent can be generally used for extraction. Or you may select and use 2 or more types. For example, although water, alcohol, glycols etc. can be mentioned, it is not limited to this. Examples of alcohols include ethanol, methanol, n-propanol and isopropanol. Examples of glycols include butylene glycol and propylene glycol. Other aqueous solvents include acetone and the like. These solvents may be used alone or as an aqueous solution, and may be used as any two or more mixed solvents.

 抽出に用いる水性溶媒の温度は、例えば、0℃以上であり、活性成分に影響を与えないのであれば特に限定されることはない。沸騰状態又は超臨界状態にある水性溶媒を使用することもできるが、5℃~600℃の水性溶媒を使用するのが好ましく、10℃~200℃の水性溶媒を使用するのがより好ましい。
 したがって、抽出用の水性溶媒とは、沸騰状態や超臨界状態にある水性溶媒も含むものである。抽出に使用する水性溶媒の量は、ユーグレナ中に含まれる水溶性活性成分を十分に溶解することができる量であることが好ましい。
The temperature of the aqueous solvent used for extraction is, for example, 0 ° C. or higher, and is not particularly limited as long as it does not affect the active ingredient. Although an aqueous solvent in a boiling or supercritical state can be used, it is preferable to use an aqueous solvent at 5 ° C. to 600 ° C., and it is more preferable to use an aqueous solvent at 10 ° C. to 200 ° C.
Therefore, the aqueous solvent for extraction also includes an aqueous solvent in a boiling state or a supercritical state. The amount of aqueous solvent used for extraction is preferably an amount capable of sufficiently dissolving the water-soluble active ingredient contained in Euglena.

 抽出方法も特に限定されず、例えば、以下に示す方法により抽出を行うことができるが、これに限定されることなく、通常の抽出方法を自由に選択して用いることができる。例えば、ユーグレナの藻体乾燥粉末を水性溶媒に所定時間浸漬した後に遠心分離又は濾過する方法、ユーグレナの藻体乾燥粉末を水性溶媒に加えて震盪して均一に分散させた後に遠心分離又は濾過する方法、などが挙げられる。
 また、抽出を促進するために、ユーグレナを添加後の水性溶媒を加熱することも可能である。
The extraction method is also not particularly limited, and, for example, the extraction can be performed by the method described below, but it is not limited thereto, and a common extraction method can be freely selected and used. For example, a method of immersing dried algal body powder of Euglena in an aqueous solvent for a predetermined time and then centrifuging or filtering it, or adding a dried powder of algal body Euglena to an aqueous solvent and shaking it uniformly for centrifugation or filtering Methods, etc.
It is also possible to heat the aqueous solvent after addition of Euglena to facilitate the extraction.

 ユーグレナの水抽出は、以下に示すような通常の方法で行うことができるが、これに限定されるものではない。例えば、ユーグレナ組織及び水を容器に入れ、適宜攪拌又は震盪しながら所定時間静置し、得られた抽出液は、そのまま水抽出物として使用可能である。また、例えば、そのような抽出液を遠心して得られる上清を水抽出物として使用することもできる。また、そのような抽出液又は上清を濃縮、乾燥して水分を除去し、これを水抽出物として使用することもできる。水抽出は、抽出効率を上げて抽出時間を短縮するために、水に、少量、例えば、10質量%以下のアルコール、好ましくはエタノールを添加して行ってもよい。
 水抽出を行う場合の抽出時間は、活性成分が抽出される時間であれば特に限定されず、数秒~数十時間の範囲で、抽出の温度に応じて適宜設定することができる。
Euglena water extraction can be carried out by the usual method as shown below, but is not limited thereto. For example, Euglena tissue and water are placed in a container and allowed to stand for a predetermined time while stirring or shaking as appropriate, and the obtained extract can be used as it is as a water extract. Also, for example, the supernatant obtained by centrifuging such extract can be used as a water extract. Alternatively, such extract or supernatant may be concentrated, dried to remove water and used as a water extract. The water extraction may be performed by adding a small amount, for example, 10% by mass or less of an alcohol, preferably ethanol, to the water in order to increase the extraction efficiency and shorten the extraction time.
The extraction time in the case of water extraction is not particularly limited as long as it is a time during which the active ingredient is extracted, and can be appropriately set according to the temperature of extraction within a range of several seconds to several tens of hours.

 熱水による抽出は、以下に示すような、通常用いられている方法で行なうことができるが、これに限定されるものではない。ユーグレナを、通常用いられる抽出器に水とともに導入した後に、加熱することで抽出を行う。沸騰水または超臨界状態にある水を使用して抽出する場合には、水の蒸気圧に耐え得る抽出器を使用する必要がある。抽出時の圧力は1~5000気圧に設定することができ、60~400気圧に設定するのが好ましい。
 高温高圧下で抽出を行なう場合には、抽出時間が長す過ぎると活性成分が分解したり、化学反応を起こすことがある。従って、高温高圧下で抽出を行なうときには、抽出時間を短時間、例えば、3分以内とするのが好ましく、1分以内とするのがより好ましく、30秒以内とすることが特に好ましい。
Extraction with hot water can be carried out by a commonly used method as shown below, but it is not limited thereto. Extraction is carried out by heating Euglana after introducing it into a commonly used extractor together with water. In the case of extraction using boiling water or water in the supercritical state, it is necessary to use an extractor capable of withstanding the water vapor pressure. The pressure at the time of extraction can be set to 1 to 5000 atm, preferably 60 to 400 atm.
In the case of extraction under high temperature and pressure, if the extraction time is too long, the active ingredient may be decomposed or a chemical reaction may occur. Therefore, when extraction is performed under high temperature and high pressure, the extraction time is preferably short, for example, 3 minutes or less, more preferably 1 minute or less, and particularly preferably 30 seconds or less.

 抽出したユーグレナ抽出物は、そのままでも本実施形態に係る筋量増加剤の有効成分として用いることができるが、当該抽出物を更に、適当な分離手段(例えば、分配抽出、ゲル濾過法、シリカゲルクロマトグラフィー、逆相若しくは順相の高速液体クロマトグラフィーなど)により活性の高い画分を分画して用いることも可能である。
 また、ユーグレナ抽出物やその画分を、濃縮、乾燥して水性溶媒を除去し、これを水性溶媒抽出物として使用することもできる。
Although the extracted Euglena extract can be used as it is as an active ingredient of the muscle mass increasing agent according to the present embodiment, the extract can be further separated by an appropriate separation means (eg, distribution extraction, gel filtration method, silica gel chromatography It is also possible to fractionate the highly active fraction by chromatography, reverse phase or normal phase high performance liquid chromatography, etc.).
Alternatively, the Euglena extract or a fraction thereof may be concentrated and dried to remove an aqueous solvent, which may be used as an aqueous solvent extract.

<筋量増加剤>
 本実施形態に係る筋量増加剤は、ユーグレナ由来物質を有効成分とする筋量増加剤(筋肉量増加剤)である。
 「ユーグレナ由来物質」には、ユーグレナ生細胞やユーグレナの乾燥藻体などのユーグレナ藻体の他、ユーグレナの水性溶媒抽出物、ユーグレナ藻体の加工品等が含まれる。
<Muscle increaser>
The muscle mass increasing agent according to the present embodiment is a muscle mass increasing agent (muscle mass increasing agent) containing an Euglena-derived substance as an active ingredient.
The “Euglena-derived substance” includes Euglena algal cells such as Euglena live cells and dried algal cells of Euglena, as well as aqueous solvent extracts of Euglena, processed products of Euglena algal bodies, and the like.

 「筋量増加剤」とは、筋量(筋肉量)を増加させる剤のことをいい、「筋肉量増加剤」と同義である。筋量の増加は、例えば、筋量計などにより筋肉量を測定することで評価することができる。 The “muscle mass increaser” refers to an agent that increases muscle mass (muscle mass) and is synonymous with “muscle mass increaser”. The increase in muscle mass can be evaluated, for example, by measuring the muscle mass using a muscle mass meter or the like.

 筋量の増加の対象となる筋は特に限定されないが、骨格筋が好ましい。骨格筋としては、例えば、胸鎖乳突筋、大胸筋、小胸筋、前鋸筋、鎖骨下筋、腹直筋、外腹斜筋、内腹斜筋、腹横筋、腰方形筋、僧帽筋、広背筋、脊柱起立筋、肩甲挙筋、菱形筋、三角筋、小円筋、棘上筋、棘下筋、肩甲下筋、大円筋、烏口腕筋、上腕二頭筋、上腕筋、腕橈骨筋、上腕三頭筋、肘筋、円回内筋、方形回内筋、回外筋、尺側手根屈筋、橈側手根屈筋、長掌筋、浅指屈筋、深指屈筋、長母指屈筋、長橈側手根伸筋、短橈側手根伸筋、尺側手根伸筋、指伸筋、示指伸筋、小指伸筋、長母指伸筋、短拇指伸筋、長母指外転筋、中様筋(4筋)、掌側骨間筋(3筋)、背側骨間筋(4筋)、小指外転筋、短小指屈筋、小指対立筋、短掌筋、母指内転筋、短拇指屈筋、母指対立筋、短拇指外転筋、大腿直筋、外側公筋、中間公筋、内側広筋、腸骨筋、大腰筋、小腰筋、縫工筋、恥骨筋、大腿筋膜張筋、大殿筋、大腿二頭筋、大腿四頭筋、半腱様筋、半膜様筋、中殿筋、小殿筋、薄筋、長内転筋、短内転筋、大内転筋、深層外旋六筋、腓腹筋、ヒラメ筋、膝窩筋、後脛骨筋、長趾屈筋、長母趾屈筋、足底筋、前脛骨筋、長腓骨筋、短腓骨筋、第3腓骨筋、長母趾伸筋、長趾伸筋などが挙げられる。 The muscle targeted for increase in muscle mass is not particularly limited, but skeletal muscle is preferred. The skeletal muscle includes, for example, chest-crest mastoid muscle, pectoralis major muscle, minor pectoral muscle, anterior serratus muscle, subclavian muscle, rectus abdominis muscle, external oblique muscle, internal oblique muscle, transverse abdominal muscle, hippocampus muscle, Cap muscles, latissimus dorsi, erector spinae muscles, elevated shoulder muscles, rhomboids, deltoids, detrusors, supraspinatus, infraspinatus, subscapularis, great fornix, brachial muscles, biceps brachii , Brachial muscle, brachial muscle, triceps brachial muscle, elbow muscle, circular pronation muscle, square pronation muscle, pronation muscle, ulnar carpal flexor muscle, dorsal carpal flexor muscle, long palm muscle, superficial flexor muscle, deep Flexor digitus longus, flexor longus thumb, extensor carpi lateralis extensors, extensor carpi lateralis carposi, ulnar carpal extensor, finger extensor, index finger extensor, finger extensor, long thumb extensor Muscle, long forefinger abductor, medial muscle (4), palmar interosseous muscle (3 for), dorsal interosseous muscle (4 for), small finger abductor, short minor flexor, minor finger alternative muscle, Short palm muscle, thumb adductor, short finger flexor, thumb alternative muscle, short finger abductor, rectus femoris, lateral public muscle, middle public muscle, medial spread , Iliac muscle, large lumbar muscle, psoas muscle, gastrocnemius muscle, pubic muscle, thigh fascia tension muscle, gluteus maximus muscle, biceps femoris muscle, quadriceps femoris muscle, semi tendon muscle, semimembranoid muscle, middle Gluteus muscle, gluteus maximus muscle, thin muscle, long adductor, short abductor, large abductor, extracorporeal external rotatory muscle, gastrocnemius, soleus muscle, popliteal muscle, posterior tibial muscle, long flexor, long mother Flexors, plantar muscles, tibialis anterior, long peroneal muscle, short peroneal muscle, third peroneal muscle, long toe extensor, long toe extensor, etc.

 大腿四頭筋は、膝関節の伸展や股関節の屈曲、具体的には、膝を伸ばして立ち上がる動作、歩行動作、走行動作に関与している。従って、大腿四頭筋の筋量が増加すると、立ち上がり、歩行、走行などの動作が向上し、ロコモティブシンドロームやサルコペニアを予防、改善することができる。 The quadriceps muscles are involved in the extension of the knee joint and the flexion of the hip joint, specifically, the action of stretching and standing the knee, the walking action, and the running action. Therefore, when the muscle mass of the quadriceps femoris increases, movements such as standing up, walking and running improve, and locomotive syndrome and sarcopenia can be prevented and improved.

 ユーグレナ由来物質は、筋肉量を増加させる作用を有しているため筋肉増強剤として用いることができる。「筋肉増強剤」とは、筋肉を増強する剤のことをいい、筋肉の増強は、例えば、筋肉量、筋力によって評価することができる。
 「筋肉を増強する」とは、筋肉量が増加することや、筋力が向上することをいう。筋肉量の増加は、例えば、筋肉量の測定から判断され、筋力の向上は、例えば、筋力計などによる筋力の測定から判断される。
The euglena-derived substance can be used as a muscle enhancing agent since it has an action to increase muscle mass. The "muscle-enhancing agent" refers to an agent that strengthens muscles, and muscle strengthening can be evaluated by, for example, muscle mass and muscle strength.
"Strengthening muscles" means increasing muscle mass and improving muscle strength. An increase in muscle mass is determined, for example, from measurement of muscle mass, and an improvement in muscle strength is determined, for example, from measurement of muscle strength by a muscle meter or the like.

<ロコモティブシンドローム予防剤、ロコモティブシンドローム治療剤、サルコペニア予防剤、サルコペニア治療剤>
 本実施形態に係る筋量増加剤は、ユーグレナ由来物質を有効成分として含有し、ロコモティブシンドロームやサルコペニアを予防又は治療するためのロコモティブシンドローム予防剤、ロコモティブシンドローム治療剤、サルコペニア予防剤、サルコペニア治療剤としても用いることができる。
 「ロコモティブシンドロームを治療する」、「サルコペニアを治療する」とは、本実施形態の筋量増加剤の有効成分であるユーグレナ由来物質を摂取する前の状態と比べて、ロコモティブシンドロームやサルコペニアの症状が軽減することをいう。
<Locomotive syndrome prophylactic agent, locomotive syndrome therapeutic agent, sarcopenia prophylactic agent, sarcopenia therapeutic agent>
The muscle mass increaser according to the present embodiment contains an Euglena-derived substance as an active ingredient, and is an agent for preventing locomotive syndrome for preventing or treating locomotive syndrome and sarcopenia, an agent for treating locomotive syndrome, an agent for preventing sarcopenia, and a treatment for sarcopenia Can also be used.
“To treat locomotive syndrome” and “to treat sarcopenia” refer to the symptoms of locomotive syndrome and sarcopenia compared to the condition before taking the euglena-derived substance which is the active ingredient of the muscle mass increasing agent of this embodiment. It means to reduce.

<運動機能維持剤、運動機能改善剤>
 本実施形態に係る筋量増加剤は、ユーグレナ由来物質を有効成分として含有し、運動機能を維持するための運動機能維持剤や、運動機能を改善するための運動機能改善剤としても用いることができる。
 「運動機能を維持する」とは、対比される状態、例えば、本実施形態の筋量増加剤の有効成分であるユーグレナ由来物質を摂取する前の状態と比べて、「立つ」、「歩く」等の運動機能が維持されることをいう。
 「運動機能を改善する」とは、対比される状態、例えば、本実施形態の筋量増加剤の有効成分であるユーグレナ由来物質を摂取する前の状態と比べて、「立つ」、「歩く」等の運動機能が改善されることをいう。
<Motor function maintenance agent, motor function improvement agent>
The muscle mass increaser according to the present embodiment contains an Euglena-derived substance as an active ingredient, and may be used also as a motor function maintenance agent for maintaining the motor function or a motor function improvement agent for improving the motor function. it can.
“Maintaining the motor function” means “stand up” or “walk” as compared with the state to be compared, for example, the state before taking the euglena-derived substance which is the active ingredient of the muscle mass increasing agent of the present embodiment. It means that motor function such as is maintained.
“Improvement of motor function” means “stand up” or “walk” as compared with the condition to be compared, for example, the condition before taking the euglena-derived substance which is the active ingredient of the muscle mass increasing agent of the present embodiment. Say that motor function such as is improved.

 <筋萎縮関連遺伝子発現抑制剤、筋萎縮予防剤及び筋タンパク質分解抑制剤>
 ユーグレナ由来物質は、筋萎縮関連遺伝子の発現を抑制する作用を有しているため、筋萎縮関連遺伝子発現抑制剤、筋萎縮予防剤、筋タンパク質分解抑制剤として用いることが可能である。
 ユーグレナ由来物質によって発現が抑制される筋萎縮関連遺伝子の例としては、例えば、ユビキチンリガーゼの遺伝子(Cbl-b(配列番号1),Atrogin-1(配列番号2),MuRF1(配列番号3))が例示される。
<A muscle atrophy-related gene expression inhibitor, muscle atrophy preventive agent and muscle protein degradation inhibitor>
Since the Euglena-derived substance has an action of suppressing the expression of a muscle atrophy-related gene, it can be used as a muscle atrophy-related gene expression inhibitor, a muscle atrophy preventive agent, and a muscle protein degradation inhibitor.
Examples of muscle atrophy-related genes whose expression is suppressed by euglena-derived substances include, for example, ubiquitin ligase genes (Cbl-b (SEQ ID NO: 1), Atrogin-1 (SEQ ID NO: 2), MuRF1 (SEQ ID NO: 3)) Is illustrated.

 <筋合成関連遺伝子発現促進剤及び筋合成促進剤>
 ユーグレナ由来物質は、筋合成関連遺伝子の発現を促進する作用を有しているため、筋合成関連遺伝子発現促進剤や筋合成促進剤として用いることが可能である。
 ユーグレナ由来物質によって発現が促進される筋合成関連遺伝子の例としては、例えば、筋タンパク質の合成を促進するS6Kの遺伝子(配列番号5)が例示される。
Muscle synthesis related gene expression promoter and muscle synthesis promoter
Euglena-derived substances have the effect of promoting the expression of muscle synthesis-related genes, and thus can be used as muscle synthesis-related gene expression promoters and muscle synthesis promoters.
As an example of a muscle synthesis related gene whose expression is promoted by the euglena-derived substance, for example, the gene of S6K (SEQ ID NO: 5) which promotes the synthesis of muscle protein is exemplified.

<用途>
 実施形態に係るユーグレナ由来物質を有効成分として含有する筋量増加剤は、筋肉量の減少や筋力が低下している患者や、ロコモティブシンドロームやサルコペニアの確定診断を受けた患者に投与される。
 また、本実施形態のユーグレナ由来物質を有効成分として含有する筋量増加剤は、医薬組成物、健康食品等の食品組成物として構成され、筋肉量の減少や筋力の低下を自覚した者や、ロコモティブシンドロームやサルコペニアに罹患する可能性の高い者等、ロコモティブシンドロームやサルコペニアの予備軍の者に、予防的に投与される。
<Use>
The muscle mass increasing agent containing the Euglena-derived substance according to the embodiment as an active ingredient is administered to a patient with decreased muscle mass or muscle strength, or a patient who has a definitive diagnosis of locomotive syndrome or sarcopenia.
Moreover, the muscle mass increasing agent containing the Euglena-derived substance of the present embodiment as an active ingredient is constituted as a food composition such as a pharmaceutical composition, a health food, etc. Prophylactically administered to people with locomotive syndrome or sarcopenia reserve forces, such as those who are likely to suffer from locomotive syndrome or sarcopenia.

 ユーグレナ藻体などのユーグレナ由来物質は、食品としても摂取可能で副作用がないため、ロコモティブシンドロームやサルコペニアの確定診断を受ける前であっても、投与可能である。 Euglena-derived substances such as Euglena algae can be taken as food and have no side effects, so they can be administered even before a definitive diagnosis of locomotive syndrome or sarcopenia.

 一般的に、筋肉量の減少や筋力の低下、ロコモティブシンドロームやサルコペニアは、生活習慣、加齢、食生活、運動不足など、様々な要因によって引き起こされることが知られている。
 よって、本実施形態のユーグレナ由来物質を有効成分として含有する筋量増加用食品組成物、筋量増加剤を、生活習慣、加齢、食生活、運動不足により、筋肉量の減少や筋力の低下する可能性の高い人、ロコモティブシンドロームやサルコペニアに罹患する可能性の高い環境にある人、例えば、生活習慣や食生活の乱れた家庭の人に対して、長期間継続投与できる。
Generally, it is known that muscle loss and muscle weakness, locomotive syndrome and sarcopenia are caused by various factors such as lifestyle, aging, diet and lack of exercise.
Therefore, the muscle mass increasing food composition containing the Euglena-derived substance of the present embodiment as an active ingredient, a muscle mass increasing agent, a decrease in muscle mass and a decrease in muscle strength due to lifestyle, aging, diet and lack of exercise. It can be administered continuously for a long period of time to people who are likely to be ill, those who are likely to suffer from locomotive syndrome or sarcopenia, for example, people at home whose lifestyle or diet has been disturbed.

 本実施形態に係るユーグレナ由来物質を有効成分として含有する筋量増加用食品組成物、筋量増加剤を投与する対象は、上記症状又は状態の者や、ヒト以外の動物に限定されるものではない。 The muscle mass increasing food composition containing the Euglena-derived substance according to the present embodiment as an active ingredient, and the target for administering the muscle mass increasing agent are limited to persons having the above condition or condition or animals other than human beings Absent.

 また、40歳以降の年齢のヒトに、特に60歳または65歳以上の高齢者にユーグレナ由来物質を有効成分として含有する筋量増加用食品組成物、筋量増加剤を投与することができる。
 40歳以上のヒト、特に60歳または65歳以上の高齢者は、加齢によって筋肉量が減少したり、筋力が低下したりする傾向があるが、ユーグレナ由来物質が備える、筋肉量増加作用により、筋肉量の減少や筋力の低下を抑制することができる。
In addition, a muscle composition for increasing muscle mass and an agent for increasing muscle mass, which contain an Euglena-derived substance as an active ingredient, can be administered to humans of 40 years of age or older, particularly to elderly people of 60 or 65 years of age.
People over the age of 40, especially elderly people over the age of 60 or 65, tend to lose muscle mass and muscle strength with age, but with the Euglena-derived substance, the muscle mass increasing action Can reduce muscle mass loss and muscle strength loss.

 本実施形態の筋量増加用食品組成物及び筋量増加剤によれば、筋肉量を増加させることができるため、筋肉量の減少や筋力の低下が関連する疾病の予防・治療や、寝たきり状態の防止、転倒による怪我の防止への応用が期待される。 According to the food composition for increasing muscle mass and muscle mass increasing agent of the present embodiment, since muscle mass can be increased, prevention and treatment of diseases associated with decrease in muscle mass and muscle strength, bed rest state It is expected to be applied to the prevention of injuries and the prevention of injuries caused by falls.

 また、本実施形態に係るユーグレナ由来物質を有効成分として含有する筋量増加剤は、薬理学的に許容され得る添加剤を加え、食品組成物、医薬組成物等の組成物等として用いることができる。 In addition, the muscle mass increasing agent containing the Euglena-derived substance according to the present embodiment as an active ingredient may be added to a pharmacologically acceptable additive and used as a composition such as a food composition, a pharmaceutical composition, etc. it can.

  (食品組成物)
 また、本実施形態のユーグレナ由来物質は、食品にも用いることが可能である。
 本実施形態の筋量増加用食品組成物、サルコペニア予防用、サルコペニア改善用、ロコモティブシンドローム予防用又はロコモティブシンドローム改善用の食品組成物、筋萎縮関連遺伝子発現抑制用、筋タンパク質分解抑制用、筋萎縮予防用、筋合成関連遺伝子発現促進用又は筋合成促進用の食品組成物は、食品の分野では、目的となる作用を有効に発揮できる有効な量のユーグレナ由来物質を食品素材として、各種食品に配合することにより、当該作用を有する食品組成物を提供することができる。
 すなわち、本発明は、食品の分野において、筋量増加用等と表示された食品の食品組成物を提供することができる。当該食品組成物としては、一般の食品のほか、特定保健用食品、栄養機能食品、機能性表示食品、病院患者用食品、サプリメント等が挙げられる。また、食品添加物として用いることもできる。
 当該食品組成物としては、例えば、調味料、畜肉加工品、農産加工品、飲料(清涼飲料、アルコール飲料、炭酸飲料、乳飲料、果汁飲料、茶、コーヒー、栄養ドリンク等)、粉末飲料(粉末ジュース、粉末スープ等)、濃縮飲料、菓子類(キャンディ(のど飴)、クッキー、ビスケット、ガム、グミ、チョコレート等)、パン、シリアル等が挙げられる。また、特定保健用食品、栄養機能食品、機能性表示食品等の場合、カプセル、トローチ、シロップ、顆粒、粉末等の形状であっても良い。
(Food composition)
In addition, the Euglena-derived substance of the present embodiment can also be used for food.
Food composition for increasing muscle mass of the present embodiment, for preventing sarcopenia, for improving sarcopenia, food composition for preventing locomotive syndrome or for improving locomotive syndrome, for suppressing muscle atrophy-related gene expression, for suppressing muscle protein degradation, muscle atrophy In the food field, food compositions for preventing, muscle synthesis-related gene expression promotion, or muscle synthesis promotion use various amounts of food materials as an effective amount of an euglena-derived substance capable of effectively exerting the intended action. By blending, it is possible to provide a food composition having the effect.
That is, the present invention can provide a food composition of food labeled as for increasing muscle mass and the like in the field of food. Examples of the food composition include, in addition to general foods, food for specified health use, nutritive function food, functional display food, food for hospital patients, supplement and the like. It can also be used as a food additive.
Examples of the food composition include seasonings, processed meat products, processed agricultural products, beverages (soft drinks, alcoholic drinks, alcoholic drinks, carbonated drinks, milk drinks, fruit drinks, tea, coffee, nutritional drinks, etc.), powdered drinks (powders) Juices, powdered soups, etc., concentrated beverages, confectionery (candy (nodice), cookies, biscuits, gums, gummi, chocolate, etc.), bread, cereals, etc. can be mentioned. Further, in the case of food for specified health use, nutritive function food, functional display food and the like, it may be in the form of capsules, troches, syrups, granules, powders and the like.

 ここで特定保健用食品とは、生理学的機能等に影響を与える保健機能成分を含む食品であって、消費者庁長官の許可を得て特定の保健の用途に適する旨を表示可能なものである。本発明においては、特定の保健用途として「自立した日常生活を送る上で必要な筋肉量及び筋力の低下抑制に役立つ」、「歩行能力の改善に役立つ」、「筋肉量や筋力の維持」、「筋肉量や筋力の維持をサポート」、「中高年齢者において加齢により衰える歩行能力の維持」、「加齢によって衰える筋肉の維持に役立つ筋肉をつくる力をサポート」、などと表示して販売される食品となる。
 また栄養機能食品とは、栄養成分(ビタミン、ミネラル)の補給のために利用される食品であって、栄養成分の機能を表示するものである。栄養機能食品として販売するためには、一日当たりの摂取目安量に含まれる栄養成分量が定められた上限値、下限値の範囲内にある必要があり、栄養機能表示だけでなく注意喚起表示等もする必要がある。
 また機能性表示食品とは、事業者の責任において、科学的根拠に基づいた機能性を表示した食品である。販売前に安全性及び機能性の根拠に関する情報などが消費者庁長官へ届け出られたものである。
Here, the food for specified health use is a food containing a health functional ingredient that affects physiological functions etc., and can be displayed as being suitable for a specific use of health with the permission of the Commissioner of the Consumer Agency. is there. In the present invention, as a specific health application, "helps to suppress the decrease in muscle mass and muscle strength necessary to live an independent daily life", "helps to improve walking ability", "maintenance of muscle mass and muscle strength", "Support for maintenance of muscle mass and muscle strength", "maintenance of walking ability that declines with age in middle-aged and elderly people", "Supporting the ability to create muscles that help maintain muscles that decline with aging," etc. Be the food to be
Also, the nutritive function food is a food used for supplementation of nutritional components (vitamins, minerals), and displays the function of the nutritional components. In order to sell it as a nutritionally functional food, it is necessary for the amount of nutritional component contained in the daily intake standard amount to be within the defined upper limit value and lower limit value, and not only the display of nutritional function but also alert display etc. You also need to
In addition, the functional labeling food is a food which displays scientific basis-based functionality at the responsibility of the business operator. Information on the basis of safety and functionality etc. was sent to the Secretary General of the Consumer Agency before sales.

 本実施形態に係る食品組成物には、ユーグレナ由来物質に加え、通常食品組成物に用いることができる成分を、1種または2種以上自由に選択して配合することが可能である。例えば、各種調味料、保存剤、乳化剤、安定剤、香料、着色剤、防腐剤、pH調整剤などの、食品分野で通常使用し得る全ての添加剤を含有させることができる。 In the food composition according to the present embodiment, in addition to the Euglena-derived substance, it is possible to freely select and mix one or two or more kinds of components that can be used for the food composition. For example, all additives which can usually be used in the food field can be contained, such as various seasonings, preservatives, emulsifiers, stabilizers, flavors, coloring agents, preservatives, pH adjusters and the like.

 本実施形態に係る食品組成物には、ユーグレナ由来物質以外に、筋肉量増加作用があることが知られている3-ヒドロキシ-3-メチルブチレート(HMB)、ブラックジンジャー由来5,7-ジメトキシフラボン、ロイシン40%配合必須アミノ酸、イミダゾールペプチド、CoQ10などの物質を1種以上添加することも可能である。 In the food composition according to the present embodiment, 3-hydroxy-3-methylbutyrate (HMB), which is known to have an action to increase muscle mass, in addition to euglena-derived substances It is also possible to add one or more substances such as flavone, leucine 40% combination essential amino acid, imidazole peptide, CoQ10 and the like.

  (医薬組成物)
 本実施形態に係るユーグレナ由来物質を有効成分として含有する筋量増加剤、ロコモティブシンドローム予防剤、ロコモティブシンドローム治療剤、サルコペニア予防剤、サルコペニア治療剤、運動機能維持剤、運動機能改善剤、筋萎縮関連遺伝子発現抑制剤及び筋タンパク質分解抑制剤、筋萎縮予防用、筋合成関連遺伝子発現促進剤、筋合成促進剤は、医薬組成物として利用することができる。
 本実施形態の筋量増加剤、ロコモティブシンドローム予防剤、ロコモティブシンドローム治療剤、サルコペニア予防剤、サルコペニア治療剤、運動機能維持剤、運動機能改善剤、筋萎縮関連遺伝子発現抑制剤、筋タンパク質分解抑制剤、筋萎縮予防剤、筋合成関連遺伝子発現促進剤、筋合成促進剤は、医薬の分野では、目的となる作用を有効に発揮できる量のユーグレナ由来物質と共に、薬学的に許容される担体や添加剤を配合することにより、当該作用を有する医薬組成物が提供される。当該医薬組成物は、医薬品であっても医薬部外品であってもよい。
 当該医薬組成物は、内用的に適用されても、また外用的に適用されても良い。従って、当該医薬組成物は、内服剤、静脈注射、皮下注射、皮内注射、筋肉注射及び/又は腹腔内注射等の注射剤、経粘膜適用剤、経皮適用剤等の製剤形態で使用することができる。
 当該医薬組成物の剤型としては、適用の形態により、適当に設定できるが、例えば、錠剤、顆粒剤、カプセル剤、粉末剤、散剤などの固形製剤、液剤、懸濁剤などの液状製剤、軟膏剤、またはゲル剤等の半固形剤が挙げられる。
(Pharmaceutical composition)
A muscle mass increasing agent containing the Euglena-derived substance according to the present embodiment as an active ingredient, a locomotive syndrome preventive agent, a locomotive syndrome therapeutic agent, a sarcopenia preventive agent, a sarcopenia therapeutic agent, a motor function maintenance agent, a motor function improving agent, muscle atrophy related Gene expression inhibitors and muscle protein degradation inhibitors, for preventing muscle atrophy, muscle synthesis related gene expression promoters, and muscle synthesis promoters can be used as pharmaceutical compositions.
The muscle mass increase agent of the present embodiment, locomotive syndrome preventive agent, locomotive syndrome therapeutic agent, sarcopenia preventive agent, sarcopenia therapeutic agent, motor function maintenance agent, motor function improvement agent, muscle atrophy related gene expression inhibitor, muscle protein degradation inhibitor In the pharmaceutical field, a muscle atrophy preventive agent, a muscle synthesis related gene expression promoter and a muscle synthesis promoter are used together with a pharmaceutically acceptable carrier or additive together with an amount of an euglena-derived substance capable of effectively exerting the intended action. By incorporating an agent, a pharmaceutical composition having the above-mentioned action is provided. The pharmaceutical composition may be a pharmaceutical or quasi-drug.
The pharmaceutical composition may be applied internally or externally. Therefore, the pharmaceutical composition is used in the form of an internal preparation, intravenous injection, subcutaneous injection, intradermal injection, injection such as intramuscular injection and / or intraperitoneal injection, transmucosal application, transdermal application and the like. be able to.
The dosage form of the pharmaceutical composition can be appropriately set depending on the form of application, for example, solid preparations such as tablets, granules, capsules, powders, powders, etc., liquid preparations such as solutions, suspensions, etc. Semisolid agents such as ointments or gels are included.

 本実施形態に係る医薬組成物には、薬学的に許容される添加剤を1種または2種以上自由に選択して含有させることができる。
 例えば、本実施形態に係る医薬組成物を経口剤に適用させる場合、例えば、賦形剤、結合剤、崩壊剤、界面活性剤、保存剤、着色剤、矯味剤、香料、安定化剤、防腐剤、酸化防止剤等の、医薬製剤の分野で通常使用し得る全ての添加剤を含有させることができる。また、ドラックデリバリーシステム(DDS)を利用して、徐放性製剤等にすることもできる。
In the pharmaceutical composition according to the present embodiment, one or more pharmaceutically acceptable additives can be freely selected and contained.
For example, when the pharmaceutical composition according to the present embodiment is applied to an oral preparation, for example, excipients, binders, disintegrants, surfactants, preservatives, coloring agents, flavoring agents, fragrances, stabilizers, preservatives All additives which can usually be used in the field of pharmaceutical preparations such as agents, antioxidants and the like can be contained. In addition, a drug delivery system (DDS) can be used to make a sustained release preparation or the like.

 本実施形態に係る医薬組成物には、ユーグレナ由来物質以外に、筋肉量増加作用があることが知られている3-ヒドロキシ-3-メチルブチレート(HMB)、ブラックジンジャー由来5,7-ジメトキシフラボン、ロイシン40%配合必須アミノ酸、イミダゾールペプチド、CoQ10などの物質を1種以上添加することも可能である。 In the pharmaceutical composition according to the present embodiment, 3-hydroxy-3-methylbutyrate (HMB), which is known to have an action to increase muscle mass other than Euglena-derived substances, 5,7-dimethoxycin derived from black ginger It is also possible to add one or more substances such as flavone, leucine 40% combination essential amino acid, imidazole peptide, CoQ10 and the like.

<用法・用量>
 本実施形態の筋量増加剤の用法としては、例えば、粉末剤、カプセル剤、錠剤、顆粒、液剤又はシロップ等によって経口投与すると良い。
 本実施形態の筋量増加剤の投与量や投与形態は、対象、病態やその進行状況、その他の条件によって適宜選択すればよい。例えば、ユーグレナ由来物質として、ユーグレナを選択し、ヒト(成人)を対象に筋肉量増加効果を得ることを目的として経口投与する場合には、一般に、ユーグレナを乾燥重量で1日当たり1~5000mg、好ましくは100~3000mg、さらに好ましくは500~2000mg程度となるように、1日に1~2回程度(朝と晩)、週に5回以上となる割合で継続的に投与するとよい。
<Usage / Dose>
As a usage of the muscle mass increasing agent of the present embodiment, for example, it may be orally administered by a powder, capsule, tablet, granule, solution, syrup or the like.
The dose and administration form of the muscle mass increasing agent of the present embodiment may be appropriately selected depending on the subject, the pathological condition, the progress of the condition, and other conditions. For example, when Euglena is selected as an Euglena-derived substance and orally administered for the purpose of obtaining a muscle mass increasing effect for human (adult), Euglena is generally 1 to 5000 mg by dry weight per day, preferably It is recommended to administer continuously once or twice a day (morning and evening), and at a rate of 5 times a week or more, so as to be about 100 to 3000 mg, more preferably about 500 to 2000 mg.

 以下、具体的実施例に基づいて本発明を具体的に説明するが、本発明はこれらに限定されるものではない。
 以下の実施例では、ユーグレナ由来物質としてユーグレナ粉末を用い、ユーグレナ粉末の継続摂取が筋肉量や筋関連遺伝子に与える影響の検討を行った。
Hereinafter, the present invention will be specifically described based on specific examples, but the present invention is not limited to these.
In the following examples, Euglena powder was used as an Euglena-derived substance, and the influence of continuous intake of Euglena powder on muscle mass and muscle-related genes was examined.

<実施例1>
 筋量増加剤として、ユーグレナ由来物質であるユーグレナ・グラシリス粉末(ユーグレナ藻体、(株)ユーグレナ製)を用いた。
Example 1
As a muscle mass increasing agent, Euglena gracilis powder (Euglena algal, manufactured by Euglena Co., Ltd.) which is a substance derived from Euglena was used.

<試験1 ユーグレナ粉末の継続摂取が筋肉量に与える影響の検討>
 試験1では、疾患モデルでないマウスにおいて、ユーグレナの摂取が筋肉量へ与える影響を明らかにすることを目的としている。4週間摂取後の大腿四頭筋および肝臓をサンプルとした。
<Test 1 Examination of the influence of continued intake of Euglena powder on muscle mass>
Study 1 aims to clarify the effect of euglena intake on muscle mass in mice that are not disease models. The quadriceps femoris muscle and liver after intake for 4 weeks were used as samples.

(試験方法)
・飼料調製
 CE-2食(日本クレアより購入)中にユーグレナ粉末を2%含むよう混合し、ユーグレナ食を調製した。CE-2食の栄養成分を表1乃至3に示す。
(Test method)
Preparation of feed Euglena diet was prepared by mixing 2% Euglena powder in a CE-2 diet (purchased from CLEA Japan). The nutritional components of the CE-2 diet are shown in Tables 1 to 3.

Figure JPOXMLDOC01-appb-T000001
Figure JPOXMLDOC01-appb-T000001

Figure JPOXMLDOC01-appb-T000002
Figure JPOXMLDOC01-appb-T000002

Figure JPOXMLDOC01-appb-T000003
Figure JPOXMLDOC01-appb-T000003

・摂食試験
 12週齢のC57BL/6J雄性マウスを平均体重が等しくなるよう3群に群分けし、それぞれコントロール群、ユーグレナ群とした(n=6)。コントロール群にはCE-2(通常食)、ユーグレナ群には2%ユーグレナ含有CE-2(ユーグレナ食)を摂食させた(表4)。4週間の摂食期間経過後、マウスから大腿四頭筋および肝臓を採取し、臓器重量を測定した。
Feeding test 12-week-old C57BL / 6J male mice were divided into three groups so that the average body weights would be equal, and used as a control group and an Euglena group (n = 6). The control group was fed with CE-2 (normal diet), and the Euglena group was fed with CE-2 (Euglena diet) containing 2% Euglena (Table 4). After a 4-week feeding period, quadriceps muscles and liver were collected from the mice and organ weights were measured.

Figure JPOXMLDOC01-appb-T000004
Figure JPOXMLDOC01-appb-T000004

(試験1の結果)
 結果を図1乃至4に示す。試験摂食時の体重変化(図1)および摂食量(図2)に有意な差は見られなかった。また、肝臓重量において各群間における有意な差は見られなかったが(図3)、大腿四頭筋重量においてコントロール群に対してユーグレナ群の筋重量が有意に高値を示した(図4)。
 したがって、ユーグレナ由来物質としてのユーグレナに筋肉量を増加させる作用があり、筋量増加剤や筋量増加用食品組成物として用いることができることがわかった。
(Result of test 1)
The results are shown in FIGS. There were no significant differences in body weight change (FIG. 1) and food intake (FIG. 2) at the time of the study. In addition, although there was no significant difference between the groups in liver weight (Fig. 3), the muscle weight of the Euglena group was significantly higher than that of the control group in the quadriceps femoris muscle weight (Fig. 4) .
Therefore, it was found that Euglena as an Euglena-derived substance has an action of increasing muscle mass, and can be used as a muscle mass increasing agent or a food composition for increasing muscle mass.

 また、大腿四頭筋は、膝関節の伸展や股関節の屈曲、具体的には、膝を伸ばして立ち上がる動作、歩行動作、走行動作に関与している。
 ユーグレナ由来物質としてのユーグレナは、大腿四頭筋の筋肉量を増加させる作用を有しており、立ち上がり、歩行、走行などの動作を向上させることができるため、ロコモティブシンドロームやサルコペニアを予防及び/又は改善するために用いることができることがわかった。
The quadriceps muscles are involved in the extension of the knee joint and the flexion of the hip joint, specifically, the action of stretching and standing the knee, the walking action, and the running action.
Euglena as an euglena-derived substance has an action of increasing the muscle mass of the quadriceps femoris muscle, and can improve movements such as standing up, walking, running and the like, thereby preventing and / or locomotive syndrome and sarcopenia. It has been found that it can be used to improve.

<試験2 ユーグレナ摂取による筋萎縮抑制作用に関する検討>
 試験2では、ユーグレナを摂取することで筋萎縮へ与える影響を明らかにすることを目的としている。
<Test 2 Examination about the muscle atrophy suppression effect by Euglena intake>
The purpose of Study 2 is to clarify the effect of taking Euglena on muscle atrophy.

(試験方法)
 C57BL/6マウス(雄性、12週齢、32匹)を平均体重が等しくなるよう群分けし、それぞれ通常飼育群(-)、尾懸垂群(C)、尾懸垂+ユーグレナ低濃度群(EL)、尾懸垂+ユーグレナ高濃度群(EH)とした。
 通常飼育群および尾懸垂群には水を、尾懸垂+ユーグレナ低濃度群(EL)および尾懸垂+ユーグレナ高濃度群(EH)にはユーグレナをそれぞれ40mg/mouseまたは80mg/mouseとなるようゾンデにより1日1回経口投与した。
 投与開始から7日後、尾懸垂試験を開始し、尾懸垂試験開始から10日後、イソフルラン麻酔下で屠殺した。腹部大動脈採血により血液を採取し、肝臓、ヒラメ筋、足底筋、腓腹筋、大腿四頭筋の各筋肉を摘出し、重量を測定した。
 足底筋および大腿四頭筋からTri reagentによりRNAを抽出後、cDNAを合成し、リアルタイムPCRにより各遺伝子(Cbl-b(配列番号1),Atrogin-1(配列番号2),MuRF1(配列番号3),4E-BP1(配列番号4),RPS6KB1(配列番号5))の発現量を測定した。内部標準としてβ-actinを用いた。
(Test method)
C57BL / 6 mice (male, 12 weeks old, 32 animals) are divided into groups so that the average body weight is equal, and the normal rearing group (-), tail suspension group (C), tail suspension + Euglena low concentration group (EL) Tail suspension + Euglena high concentration group (EH).
Water is used for the normal rearing group and tail suspension group, and the euglena is 40 mg / mouse or 80 mg / mouse for tail suspension + low Euglena concentration (EL) and tail suspension + high euglena concentration (EH), respectively. It was orally administered once a day.
The tail suspension test was started 7 days after the start of administration, and 10 days after the start of the tail suspension test, it was sacrificed under isoflurane anesthesia. Blood was collected by abdominal aortic blood collection, and each muscle of liver, soleus muscle, plantar muscle, gastrocnemius muscle, and quadriceps femoris muscle was removed and weighed.
After extracting RNA from the plantar and quadriceps muscles with Tri reagent, cDNA is synthesized, and each gene (Cbl-b (SEQ ID No. 1), Atrogin-1 (SEQ ID No. 2), MuRF1 (SEQ ID No.) is synthesized by real-time PCR. 3) The expression levels of 4E-BP1 (SEQ ID NO: 4) and RPS6KB1 (SEQ ID NO: 5) were measured. Β-actin was used as an internal standard.

(群分け)
 群分けは、以下のとおりである(各群n=8)。
・通常飼育群(-):尾懸垂無し、dHO投与
・尾懸垂群(C):尾懸垂有り、dHO投与
・尾懸垂+ユーグレナ低濃度群(EL):尾懸垂有り、ユーグレナ40mg/mouse/day(1%摂食相当)投与
・尾懸垂+ユーグレナ高濃度群(EH):尾懸垂有り、ユーグレナ80mg/mouse/day(2%摂食相当)投与
(Grouping)
Grouping is as follows (each group n = 8).
-Normal rearing group (-): no tail suspension, dH 2 O administration-tail suspension group (C): tail suspension, dH 2 O administration, tail suspension + Euglena low concentration group (EL): tail suspension, Euglena 40 mg / Mouse / day (1% feeding equivalent) administration · tail suspension + Euglena high concentration group (EH): tail suspension, Euglena 80 mg / mouse / day (2% feeding equivalent) administration

(試験2の結果)
(A.体重および肝臓重量測定)
 体重および肝臓重量測定の結果を図5に示す。
 試験終了時の体重は通常飼育群(-)と比較して、尾懸垂群(C)ならびに尾懸垂+ユーグレナ低濃度群(EL)に有意な減少がみられた。また、肝臓重量においても体重と同じ挙動を示した。
(Result of trial 2)
(A. Weight and liver weight measurement)
The results of body weight and liver weight measurements are shown in FIG.
Body weight at the end of the study was significantly reduced in the tail suspension group (C) and in the tail suspension + low Euglena concentration group (EL) compared to the normal rearing group (-). In addition, liver weight showed the same behavior as body weight.

(B.筋重量測定)
 筋重量測定の結果を図6及び7に示す。
 図6に示すように、通常飼育群と比較し、尾懸垂において筋重量の低下が見られたことから、尾懸垂により廃用性筋萎縮が誘導されたことが確認された。
 図7に示すように、足底筋および大腿四頭筋においてユーグレナの摂取により筋重量の低下が抑制された。ユーグレナには多くの成分が含まれており、筋タンパク質の分解阻害あるいは合成促進に寄与した可能性が考えられる。
(B. muscle weight measurement)
The results of muscle weight measurements are shown in FIGS.
As shown in FIG. 6, a decrease in muscle weight was observed in the tail suspension compared to the normal rearing group, and it was confirmed that the tail suspension induced disuse muscle atrophy.
As shown in FIG. 7, in the plantar and quadriceps muscles, Euglena consumption inhibited the decrease in muscle weight. Euglena contains many components, which may have contributed to the inhibition of muscle protein degradation or the promotion of synthesis.

 筋重量測定の結果から、ユーグレナを足底筋や大腿四頭筋の筋重量の低下を抑制するために用いることが可能であることがわかった。足底筋や大腿四頭筋の筋量が低下すると、ロコモティブシンドロームやサルコペニアの発症につながるため、ユーグレナをロコモティブシンドロームやサルコペニアの治療剤や予防剤として用いることが可能である。
 また、足底筋や大腿四頭筋は運動機能と強く関連する筋肉であるため、ユーグレナを運動機能維持剤や運動機能改善剤として用いることが可能である。
From the results of muscle weight measurement, it was found that Euglena can be used to suppress the decrease in muscle weight of the plantar and quadriceps muscles. Euglena can be used as a therapeutic agent or prophylactic agent for locomotive syndrome or sarcopenia, because reduction of muscle mass of plantar muscles and quadriceps leads to development of locomotive syndrome and sarcopenia.
In addition, since the plantar muscles and quadriceps muscles are muscles that are strongly associated with motor function, it is possible to use Euglena as a motor function maintenance agent or a motor function improvement agent.

(C.筋萎縮関連遺伝子発現量測定)
 筋萎縮関連遺伝子発現量測定の結果を図8に示す。
 図8に示すように、足底筋においてユーグレナの摂取により筋タンパク質の分解に関わるユビキチンリガーゼの遺伝子(Cbl-b(配列番号1),Atrogin-1(配列番号2),MuRF1(配列番号3)、図9)の発現を抑制する傾向が見られた。
 一方、大腿四頭筋においてユーグレナの摂取によりCbl-b(配列番号1)およびAtrogin-1(配列番号2)の遺伝子発現を抑制する傾向は見られなかった。以上の結果より、筋肉の部位によりユーグレナによる効能に差がみられる可能性が示された。
(C. Measurement of muscle atrophy-related gene expression level)
The result of muscle atrophy-related gene expression level measurement is shown in FIG.
As shown in FIG. 8, ubiquitin ligase genes (Cbl-b (SEQ ID NO: 1), Atrogin-1 (SEQ ID NO: 2), MuRF1 (SEQ ID NO: 3) involved in the degradation of muscle proteins by ingestion of euglena in plantar muscles) , Tend to suppress the expression of FIG.
On the other hand, there was no tendency to suppress the gene expression of Cbl-b (SEQ ID NO: 1) and Atrogin-1 (SEQ ID NO: 2) by ingestion of euglena in the quadriceps femoris muscle. From the above results, it has been shown that there may be differences in the efficacy of Euglena depending on the muscle site.

 筋萎縮関連遺伝子発現量測定の結果から、ユーグレナを、筋タンパク質の分解に関わるユビキチンリガーゼの遺伝子(Cbl-b(配列番号1),Atrogin-1(配列番号2),MuRF1(配列番号3))が発現することを抑制するための筋萎縮関連遺伝子発現抑制剤として用いることができることがわかった。
 また、ユーグレナを、筋タンパク質の分解を抑制する筋タンパク質分解抑制剤や、筋萎縮を予防する筋萎縮予防剤として用いることが可能であることがわかった。
From the results of measurement of muscle atrophy-related gene expression levels, Euglena was selected from the ubiquitin ligase genes (Cbl-b (SEQ ID NO: 1), Atrogin-1 (SEQ ID NO: 2), MuRF1 (SEQ ID NO: 3)) involved in the degradation of muscle proteins. It has been found that it can be used as a muscle atrophy-related gene expression inhibitor for suppressing the expression of
In addition, it was found that Euglena can be used as a muscle protein degradation inhibitor that inhibits the degradation of muscle protein and a muscle atrophy preventive agent that prevents muscle atrophy.

(D.筋合成関連遺伝子発現量測定)
 筋合成関連遺伝子発現量測定の結果を図10に示す。
 図10に示すように、足底筋と大腿四頭筋においてユーグレナの摂取により筋タンパク質の合成阻害因子である4E-BP1(配列番号4)(図11)の発現を抑制し、筋タンパク質の合成を促進するS6K(RPS6KB1、配列番号5)の遺伝子(図11)の発現を促進する傾向が見られた。ユーグレナは筋タンパク質の分解を抑制するとともに、筋合成を促進することで筋萎縮改善作用を発揮することが示唆された。また、ユーグレナには数多くの成分が含まれていることから、その作用点は複数存在し、組み合わさることで効果を発揮している可能性が考えられる。
(D. Muscle synthesis related gene expression level measurement)
The result of muscle synthesis related gene expression level measurement is shown in FIG.
As shown in FIG. 10, in the plantar and quadriceps muscles, the expression of 4E-BP1 (SEQ ID NO: 4) (FIG. 11), a muscle protein synthesis inhibitor, is suppressed by intake of euglena and the synthesis of muscle protein There was a tendency to promote the expression of the gene for S6K (RPS6KB1, SEQ ID NO: 5) which promotes It has been suggested that Euglena suppresses muscle protein degradation and promotes muscle atrophy by promoting muscle synthesis. In addition, since Euglena contains many components, there may be a plurality of action points, which may be effective when combined.

 筋合成関連遺伝子発現量測定の結果から、ユーグレナを、筋合成関連遺伝子の発現を促進する筋合成関連遺伝子発現促進剤として用いることが可能であることがわかった。
 また、ユーグレナを、筋合成を促進する筋合成促進剤として用いることが可能であることがわかった。
From the results of measurement of muscle synthesis related gene expression levels, it was found that Euglena can be used as a muscle synthesis related gene expression promoter that promotes the expression of muscle synthesis related genes.
Moreover, it turned out that it is possible to use Euglena as a muscle synthesis promoter which promotes muscle synthesis.

Claims (17)

 ユーグレナ由来物質を有効成分として含有することを特徴とする筋量増加用食品組成物。 A muscle composition for increasing muscle mass, which comprises an Euglena-derived substance as an active ingredient.  前記ユーグレナ由来物質がユーグレナであることを特徴とする請求項1に記載の筋量増加用食品組成物。 The food composition for increasing muscle mass according to claim 1, wherein the Euglena-derived substance is Euglena.  大腿四頭筋の量を増加させるために用いられることを特徴とする請求項1又は2に記載の筋量増加用食品組成物。 The food composition for increasing muscle mass according to claim 1 or 2, which is used to increase the amount of quadriceps.  ユーグレナ由来物質を有効成分として含有することを特徴とする筋量増加剤。 A muscle mass increasing agent comprising an Euglena-derived substance as an active ingredient.  ユーグレナ由来物質を有効成分として含有し、
 サルコペニア予防用、サルコペニア改善用、ロコモティブシンドローム予防用又はロコモティブシンドローム改善用の食品組成物。
Contains an Euglena-derived substance as an active ingredient,
A food composition for preventing sarcopenia, for improving sarcopenia, for preventing locomotive syndrome or for improving locomotive syndrome.
 ユーグレナ由来物質を有効成分として含有することを特徴とするロコモティブシンドローム予防剤。 A locomotive syndrome preventive agent comprising an Euglena-derived substance as an active ingredient.  ユーグレナ由来物質を有効成分として含有することを特徴とするロコモティブシンドローム治療剤。 A locomotive syndrome therapeutic agent characterized by containing an Euglena-derived substance as an active ingredient.  ユーグレナ由来物質を有効成分として含有することを特徴とするサルコペニア予防剤。 An agent for preventing sarcopenia, which comprises an Euglena-derived substance as an active ingredient.  ユーグレナ由来物質を有効成分として含有することを特徴とするサルコペニア治療剤。 A sarcopenia therapeutic agent comprising an Euglena-derived substance as an active ingredient.  ユーグレナ由来物質を有効成分として含有することを特徴とする運動機能維持剤。 A motor function maintenance agent characterized by containing an Euglena-derived substance as an active ingredient.  ユーグレナ由来物質を有効成分として含有することを特徴とする運動機能改善剤。 A motor function improving agent comprising an Euglena-derived substance as an active ingredient.  ユーグレナ由来物質を有効成分として含有し、
 筋萎縮関連遺伝子発現抑制用、筋タンパク質分解抑制用、筋萎縮予防用、筋合成関連遺伝子発現促進用又は筋合成促進用の食品組成物。
Contains an Euglena-derived substance as an active ingredient,
A food composition for suppressing muscle atrophy-related gene expression, for suppressing muscle protein degradation, for preventing muscle atrophy, for promoting muscle synthesis-related gene expression, or for promoting muscle synthesis.
 ユーグレナ由来物質を有効成分として含有し、
 筋萎縮関連遺伝子の発現を抑制するために用いられることを特徴とする筋萎縮関連遺伝子発現抑制剤。
Contains an Euglena-derived substance as an active ingredient,
A muscle atrophy-related gene expression inhibitor, which is used for suppressing the expression of a muscle atrophy-related gene.
 ユーグレナ由来物質を有効成分として含有し、
 筋タンパク質の分解を抑制するために用いられることを特徴とする筋タンパク質分解抑制剤。
Contains an Euglena-derived substance as an active ingredient,
A muscle protein degradation inhibitor, which is used to suppress the degradation of muscle protein.
 ユーグレナ由来物質を有効成分として含有し、
 筋肉の萎縮を予防するために用いられることを特徴とする筋萎縮予防剤。
Contains an Euglena-derived substance as an active ingredient,
An agent for preventing muscle atrophy, which is used to prevent muscle atrophy.
 ユーグレナ由来物質を有効成分として含有し、
 筋合成関連遺伝子の発現を促進するために用いられることを特徴とする筋合成関連遺伝子発現促進剤。
Contains an Euglena-derived substance as an active ingredient,
A muscle synthesis related gene expression promoter which is used to promote the expression of a muscle synthesis related gene.
 ユーグレナ由来物質を有効成分として含有し、
 筋合成を促進するために用いられることを特徴とする筋合成促進剤。
Contains an Euglena-derived substance as an active ingredient,
A muscle synthesis promoter characterized in that it is used to promote muscle synthesis.
PCT/JP2018/028756 2017-08-01 2018-07-31 Food composition for increasing muscle mass, muscle mass increasing agent, therapeutic agent for locomotive syndrome, therapeutic agent for sarcopenia, exercise function maintenance agent, muscle atrophy-related gene expression inhibitor, muscle protein degradation inhibitor, muscle atrophy prevention agent, muscle synthesis-related gene expression promoter, and muscle synthesis promoter Ceased WO2019026930A1 (en)

Priority Applications (1)

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CN201880049823.XA CN111031812A (en) 2017-08-01 2018-07-31 Food composition for increasing muscle mass, muscle mass increasing agent, therapeutic agent for dyskinesia syndrome, agent for treating sarcopenia, agent for maintaining motor function, agent for inhibiting expression of gene related to muscular atrophy, agent for inhibiting decomposition of muscle protein, agent for preventing muscular atrophy, agent for promoting expression of gene related to muscular synthesis, and agent for promoting muscle synthesis

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JP2017-149413 2017-08-01
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EP4209579A4 (en) * 2020-09-02 2025-01-08 Nissin Foods Holdings Co., Ltd. BIFIDOBACTERIUM TO INHIBIT THE EXPRESSION OF THE MUSCLE ATROPHY GENE

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EP4209579A4 (en) * 2020-09-02 2025-01-08 Nissin Foods Holdings Co., Ltd. BIFIDOBACTERIUM TO INHIBIT THE EXPRESSION OF THE MUSCLE ATROPHY GENE

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