WO2020203792A1 - Composition destinée à convertir un muscle squelettique en muscle lent - Google Patents

Composition destinée à convertir un muscle squelettique en muscle lent Download PDF

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Publication number
WO2020203792A1
WO2020203792A1 PCT/JP2020/014079 JP2020014079W WO2020203792A1 WO 2020203792 A1 WO2020203792 A1 WO 2020203792A1 JP 2020014079 W JP2020014079 W JP 2020014079W WO 2020203792 A1 WO2020203792 A1 WO 2020203792A1
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composition
active ingredient
bifidobacterium
muscle
cells
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PCT/JP2020/014079
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English (en)
Japanese (ja)
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一弥 戸田
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森永乳業株式会社
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Priority to JP2021512011A priority Critical patent/JP7261867B2/ja
Publication of WO2020203792A1 publication Critical patent/WO2020203792A1/fr
Priority to JP2023005552A priority patent/JP2023033532A/ja

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/745—Bifidobacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A61P21/06—Anabolic agents

Definitions

  • the present invention relates to a composition that can be used for a specific purpose, such as a composition for slowing skeletal muscle.
  • Muscles are the basis of daily activities such as standing, walking, and maintaining posture. Therefore, in order to maintain the quality of life, there is a desire to maintain muscle mass as much as possible and increase muscle mass if possible, regardless of age or sex. In addition, humans and athletes (for example, athletes, sports enthusiasts, athletes, etc.) who actively prefer exercise are required to increase their muscle mass as much as possible in order to improve their athletic performance.
  • QOL quality of life
  • flail a state between a healthy state and a state requiring nursing care
  • locomotive syndrome locomotive syndrome
  • Muscle is composed of bundles of muscle fibers. Muscle fibers are classified into slow muscle fibers (Type I) and fast muscle fibers (Type II) based on properties such as contraction tension, contraction speed, and fatigue tolerance. Muscles with many slow muscle fibers have the property of having low contraction force and contraction rate but are less likely to fatigue, and contribute to endurance. On the other hand, muscles with many fast muscle fibers have a large contraction force and contraction speed, but are prone to fatigue. In addition, muscles with many slow muscle fibers act as anti-gravity muscles. In patients with disuse muscular atrophy, the muscles change from slow muscles to fast muscles, and the fast muscles are more likely to atrophy due to inactivity than the slow muscles, which interferes with daily activities such as difficulty walking and bedridden. It leads to a state of atrophy. Maintaining or strengthening slow muscles can prevent or ameliorate lack of exercise and difficulty in activities of daily living by maintaining endurance and strength.
  • Ginger extract and the like are known as substances that can promote slowing of skeletal muscle (Patent Document 1).
  • Non-Patent Document 1 It has been reported that the administration of Bifidobacterium breve B-3 increases muscle mass.
  • An object of the present invention is to provide a composition that can be used for a specific purpose, such as a composition for slowing skeletal muscle.
  • One aspect of the present invention is a composition for slowing skeletal muscle, which contains the following component (A) as an active ingredient: (A) Bifidobacterium and / or the product of the bacterium.
  • One aspect of the present invention is an endurance-enhancing composition containing the following component (A) as an active ingredient: (A) Bifidobacterium and / or the product of the bacterium.
  • One aspect of the present invention is a composition for preventing, ameliorating, and / or treating symptoms associated with muscular atrophy and / or weakness, which comprises the following component (A) as an active ingredient: (A) Bifidobacterium and / or the product of the bacterium.
  • the composition is a pharmaceutical composition. It is preferable that the composition is a food and drink composition.
  • the component (A) is the bacterium.
  • the composition preferably contains the killed cells of the bacterium as the component (A).
  • the bacterium is Bifidobacterium breve.
  • the composition preferably comprises the bacterium Bifidobacterium breve FERM BP-11175.
  • One aspect of the present invention is a method of slowing skeletal muscle, which comprises the step of administering the following component (A) to a subject: (A) Bifidobacterium and / or the product of the bacterium.
  • One aspect of the invention is a method of preventing, ameliorating, and / or treating symptoms associated with muscular atrophy and / or weakness, comprising the step of administering the following component (A) to a subject: (A) Bifidobacterium and / or the product of the bacterium.
  • One aspect of the invention is the use of the following component (A) for the prevention, amelioration, and / or treatment of symptoms associated with muscular atrophy and / or weakness: (A) Bifidobacterium and / or the product of the bacterium.
  • One aspect of the present invention is the use of the following component (A) for slowing skeletal muscle: (A) Bifidobacterium and / or the product of the bacterium.
  • One aspect of the present invention is the use of the following component (A) for the production of the composition: (A) Bifidobacterium and / or the product of the bacterium.
  • Active ingredient In the present invention, the following ingredient (A) is used as an active ingredient: (A) Bifidobacterium and / or the product of the bacterium.
  • the above component (A) is also referred to as an "active ingredient".
  • the skeletal muscle By using the active ingredient, specifically, by administering the active ingredient to the subject, the skeletal muscle can be slowed down in the subject, that is, the effect of slowing the skeletal muscle in the subject is achieved. can get.
  • This effect is also called "skeletal muscle slowing effect”.
  • Skeletal muscles include trunk muscles (ie, trunk skeletal muscles) and limb muscles (ie, upper and lower limb skeletal muscles). Examples of the skeletal muscle of the lower limbs include the gastrocnemius muscle and the soleus muscle.
  • the skeletal muscle slowing effect may be obtained in all skeletal muscles (that is, whole body skeletal muscles) or in some skeletal muscles.
  • the skeletal muscle slowing effect may be obtained, for example, at a site having a high ratio of slow muscles, may be obtained at a site having a high ratio of fast muscles, or may be obtained at both of them.
  • the skeletal muscle slowing effect may be obtained, for example, at least in the gastrocnemius and / or soleus muscles.
  • “Skeletal muscle slowing” means an increase in the amount of slow muscle in skeletal muscle. In other words, "skeletal muscle slowing" means that the amount of slow muscle in the skeletal muscle is larger when the active ingredient is administered than when the active ingredient is not administered. “At the time of administration of the active ingredient” may mean after sufficient administration of the active ingredient, specifically, after administration of the active ingredient under the conditions described in the method of the present invention described later. May mean.
  • “When the active ingredient is not administered” may simply mean before administration of the active ingredient. However, “when the active ingredient is not administered” may mean, to be precise, a hypothetical condition that the same period as the administration period of the active ingredient has elapsed without administration of the active ingredient. That is, for example, when the amount of slow muscle in skeletal muscle is reduced by non-administration of the active ingredient, "skeletal muscle slowing" means that the degree of the reduction is reduced by administration of the active ingredient. Sufficiently, it is not necessary that the amount of slow muscle in the skeletal muscle is large after the administration of the active ingredient as compared with before the administration of the active ingredient. Such cases include cases where the onset or progression of slow muscle atrophy is alleviated by administration of the active ingredient.
  • the "amount of slow muscle in skeletal muscle” may mean the absolute amount of slow muscle in skeletal muscle, that is, the relative amount of slow muscle in skeletal muscle (that is, the amount of slow muscle relative to the amount of skeletal muscle). It may mean ratio) or both of them. That is, the use of the active ingredient may increase the absolute and / or relative amount of slow muscle. The use of the active ingredient may at least increase the relative amount of slow muscle. The total amount of skeletal muscle may increase as the amount of slow muscle increases. Examples of the amount of slow muscle include the number of slow muscle fibers, the cross-sectional area of slow muscle fibers, and the volume of slow muscle fibers.
  • slow muscle fiber means Type I muscle fiber.
  • Utilization of the active ingredient may increase, for example, one or more of these parameters. Utilization of the active ingredient may increase, for example, at least the number of slow muscle fibers.
  • the amount of slow muscle can be measured, for example, by a known method. Methods for measuring the amount of slow muscle include muscle biopsy, magnetic resonance imaging (MRI), and field tests.
  • the ratio of the number of slow muscle fibers to the number of muscle fibers in skeletal muscle can be measured, for example, by the method described in Examples.
  • the amount of slow muscle in skeletal muscle at the time of administration of the active ingredient is, for example, 101% or more, 103% or more, 105% or more, 110% or more, 115% or more when the non-administration of the active ingredient is taken as 100%. , 120% or more, 125% or more, or 130% or more.
  • slowing of skeletal muscle can be measured, for example, by improving fatigue tolerance, improving oxidase activity in skeletal muscle, or improving the number of mitochondria in skeletal muscle.
  • AMP-activated protein kinase activation of AMP-activated protein kinase
  • AMPK activation promoting effect Activation of AMPK may be promoted, especially in skeletal muscle.
  • a skeletal muscle slowing effect may be obtained by promoting the activation of AMPK.
  • Promoter activation of AMPK means an increase in the amount of active AMPK (ie, phosphorylated AMPK).
  • “promotion of AMPK activation” means that the amount of active AMPK is large when the active ingredient is administered as compared with when the active ingredient is not administered.
  • “Amount of activated AMPK” may mean a value standardized based on the total amount of AMPK, unless otherwise specified.
  • the amount of active AMPK can be measured, for example, by a known method.
  • Western blotting is an example of a method for measuring the amount of activated AMPK. Specifically, the amount of activated AMPK can be measured, for example, by the method described in Examples.
  • the amount of active AMPK at the time of administration of the active ingredient is, for example, 101% or more, 103% or more, 105% or more, 110% or more, 115% or more, 120 when the non-administration of the active ingredient is 100%. It may be% or more, 125% or more, or 130% or more.
  • the expression of the peroxisome growth factor activating receptor ⁇ -conjugated factor-1 ⁇ may be promoted in the subject by using the active ingredient, specifically by administering the active ingredient to the subject. That is, the effect of promoting the expression of PGC-1 ⁇ in the subject may be obtained. This effect is also referred to as "PGC-1 ⁇ expression promoting effect”. Expression of PGC-1 ⁇ may be promoted, especially in skeletal muscle. A skeletal muscle slowing effect may be obtained by promoting the expression of PGC-1 ⁇ . “Promotion of expression of PGC-1 ⁇ ” means an increase in the expression level of PGC-1 ⁇ .
  • the expression level of PGC-1 ⁇ means that the expression level of PGC-1 ⁇ is higher when the active ingredient is administered than when the active ingredient is not administered.
  • the "expression level of PGC-1 ⁇ ” may mean a value standardized based on the expression level of a housekeeping gene such as GAPDH.
  • the expression level of PGC-1 ⁇ can be measured, for example, by a known method for measuring the expression level of a gene. Examples of the method for measuring the expression level of a gene include a method for measuring the amount of mRNA and a method for measuring the amount of protein. Specific examples of the method for measuring the expression level of a gene include Northern hybridization, microarray, RT-PCR, RNA-seq, and Western blotting.
  • the expression level of PGC-1 ⁇ can be measured, for example, by the method described in Examples.
  • the expression level of PGC-1 ⁇ at the time of administration of the active ingredient is, for example, 101% or more, 103% or more, 105% or more, 110% or more, 115% or more, assuming that the non-administration of the active ingredient is 100%. It may be 120% or more, 125% or more, or 130% or more.
  • an effect based on slowing of skeletal muscle may be obtained in the subject.
  • the endurance of the subject can be improved by slowing down the skeletal muscle, for example. That is, as an effect based on the slowing of the skeletal muscle, there is an effect of improving the endurance of the subject (endurance improving effect).
  • “Endurance” means endurance for physical exercise. That is, “endurance” may specifically mean the ability to sustain physical activity. Physical exercise is not particularly limited as long as it is a movement that puts a load on muscles. Physical exercise includes sports, physical training, physical labor, and activities of daily living.
  • the muscle strength of the target can be improved by slowing down the skeletal muscle.
  • the muscle strength of the target can be improved. That is, as an effect based on the slowing of the skeletal muscle, there is an effect of improving the muscle strength of the target (muscle strength improving effect).
  • Skeletal muscle slowing is expected to prevent, ameliorate, and / or treat, for example, symptoms associated with muscle atrophy and / or weakness. That is, effects based on slowing skeletal muscle include the effect of preventing, ameliorating, and / or treating symptoms associated with muscle atrophy and / or weakness.
  • Muscle atrophy refers to loss of muscle mass. Muscle atrophy includes atrophy of skeletal muscle. Specific examples of muscular atrophy include atrophy of slow and / or fast muscles. Muscle atrophy includes, in particular, slow muscle atrophy. Muscle atrophy may, for example, reduce muscle strength. In other words, weakness may be due, for example, to muscle atrophy. Symptoms associated with muscle atrophy and / or weakness may or may not be disease.
  • Symptoms associated with muscular atrophy and / or weakness may develop or progress due to any cause, such as genetic factors, acquired factors, or aging. Symptoms associated with muscular atrophy and / or weakness include those caused by muscular atrophy and / or weakness and those associated with muscular atrophy and / or weakness. Symptoms related to muscular atrophy and / or muscle weakness include muscular atrophy itself and muscle weakness itself. Symptoms associated with muscular atrophy and / or weakness include, specifically, disused muscular atrophy, neurogenic muscular atrophy, and myogenic muscular atrophy. Disuse muscular atrophy includes age-related muscular atrophy (sarcopenia) and muscular atrophy due to insufficient muscle use. Insufficient use of muscles may be due to bedridden, weightlessness, weightless flight, body fixation, and the like.
  • Examples of body fixation include limb fixation performed in the treatment of injuries and the like.
  • As neurogenic muscular atrophy amyotrophic lateral sclerosis (ALS), primary lateral sclerosis (PLS), spinal muscular atrophy (SMA), Examples include spinal and bulbar muscular atrophy (SBMA), acute poliomyelitis (poliomyelitis), and Guillan-Barre syndrome (GBS).
  • Examples of myogenic muscular atrophy include muscular dystrophy and polymyositis (PM).
  • myasthenia gravis (MG), cachexia, frailty, locomotive syndrome (locomotive syndrome;) locomotive syndrome is also mentioned.
  • the active ingredient may or may not be used in combination with other techniques for obtaining the above-exemplified effects such as skeletal muscle slowing effect.
  • Such techniques include performing physical exercises other than activities of daily living.
  • Physical exercises other than activities of daily living include sports and physical training.
  • the present invention also provides the use of the active ingredient in applications associated with the above-exemplified effects such as skeletal muscle slowing effect. That is, the present invention uses, for example, the use of an active ingredient for slowing skeletal muscle, the use of an active ingredient for promoting the activation of AMPK, and the use of an active ingredient for promoting the expression of PGC-1 ⁇ .
  • Use of active ingredients for improving endurance Use of active ingredients for improving muscle strength, Active ingredients for prevention, improvement, and / or treatment of symptoms associated with muscle atrophy and / or muscle weakness May be offered for use.
  • the present invention also provides, for example, a composition for skeletal muscle slowing, a composition for promoting AMPK activation, a composition for promoting PGC-1 ⁇ expression, prevention and amelioration of symptoms related to muscle atrophy and / or muscle weakness. And / or the use of active ingredients for the production of compositions of the invention, such as therapeutic compositions, may be provided.
  • the present invention also provides an active ingredient for use in applications associated with the above-exemplified effects such as skeletal muscle slowing effect. That is, in the present invention, for example, an active ingredient for use in slowing skeletal muscle, an active ingredient for promoting activation of AMPK, an active ingredient for promoting expression of PGC-1 ⁇ , and endurance.
  • the active ingredient for use in improving muscle strength, the active ingredient for use in improving muscle strength, and the active ingredient for use in the prevention, amelioration, and / or treatment of symptoms associated with muscle atrophy and / or muscle weakness may be provided. ..
  • the present invention also provides, for example, a composition for skeletal muscle slowing, a composition for promoting AMPK activation, a composition for promoting PGC-1 ⁇ expression, prevention and amelioration of symptoms related to muscle atrophy and / or muscle weakness. And / or active ingredients for use in the production of the compositions of the invention, such as therapeutic compositions, may be provided.
  • the active ingredient may be used for therapeutic purposes or may be used for non-therapeutic purposes. That is, all of the above-exemplified effects may be obtained for therapeutic purposes or may be obtained for non-therapeutic purposes, unless otherwise specified.
  • the "therapeutic purpose” may mean, for example, including a treatment act on the human body for treatment, and may mean that it is performed as a medical act in particular.
  • the term "non-therapeutic purpose” may mean, for example, not including the act of treating the human body for treatment, and in particular, may mean that it is performed as a non-medical act.
  • Non-therapeutic purposes include purposes such as health promotion and beauty.
  • Prevention of symptoms may mean, for example, prevention or delay of the onset of symptoms, or reduction of the possibility of onset of symptoms.
  • Symptom improvement or “symptom treatment” may mean, for example, improvement of symptoms, prevention or delay of exacerbation of symptoms, or prevention or delay of progression of symptoms.
  • the term “improvement of symptoms” may be a general term for these events, which are not intended for treatment.
  • Treatment of symptoms may be a general term for these events and those for the purpose of treatment.
  • Bifidobacterium spp. are not particularly limited as long as a desired effect such as a skeletal muscle slowing effect can be obtained.
  • bacterium of the genus Bifidobacterium one kind of bacterium may be used, or two or more kinds of bacteria may be used in combination.
  • Bifidobacterium genus bacteria include Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium bifidum, and Bifidobacterium addressntis (Bifidobacterium addressuntis).
  • Bifidobacterium adolescentis Bifidobacterium angulatum, Bifidobacterium dentium, Bifidobacterium pseudocatenulatum, Bifidobacterium animalis ), Bifidobacterium pseudolongum, Bifidobacterium thermophilum, and the like.
  • Bifidobacterium spp. are particularly Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium bifidum, and Bifidobacterium adre.
  • Bifidobacterium adolescentis Bifidobacterium dentium, Bifidobacterium animalis, Bifidobacterium pseudolongum, Bifidobacterium thermophilum ).
  • Bifidobacterium genus bacteria include, more particularly, Bifidobacterium breve and Bifidobacterium longum.
  • Bifidobacterium breve is mentioned as a bacterium of the genus Bifidobacterium, and more particularly, Bifidobacterium breve.
  • Bifidobacterium longum includes Bifidobacterium longum subspecies longum (B. longum subsp. Longum) and Bifidobacterium longum subspecies infantis (B. longum subsp. Infantis). , Bifidobacterium longum subspecies swiss (B. longum subsp. Suis), etc., strains classified into any subspecies of Bifidobacterium longum are also included.
  • Bifidobacterium animalis also includes strains classified into any subspecies of Bifidobacterium animalis, such as Bifidobacterium animalis subspecies lactis (B. animalis subsp. Lactis). To.
  • Bifidobacterium pseudolongum includes Bifidobacterium pseudolongum subsp. Globosum and Bifidobacterium pseudolongum subspecies pseudolongum (B. pseudolongum subspecies globosum). Includes strains classified into any subspecies of bifidobacteria and pseudolongum, such as. Pseudolongum subsp. Pseudolongum).
  • Bifidobacterium breve specifically, M-16V (NITE BP-02622), MCC1274 (FERM BP-11175), ATCC 15700, B632 (DSM 24706), Bb99 (DSM 13692), ATCC 15698, DSM 24732, UCC2003, YIT4010, YIT4064, BBG-001, BR-03, C50, R0070 can be mentioned.
  • Bifidobacterium breve is particularly FERM BP-11175.
  • BB536 NITE BP-02621
  • ATCC 15697 ATCC 15707, ATCC 25962, ATCC 15702, ATCC 27533, M-63, BG7, DSM 24736, SBT 2928, NCC 490.
  • CNCM I-2170 NCC 2705 (CNCM I-2618).
  • Bifidobacterium Bifidum examples include ATCC 29521, OLB6378, and BF-1.
  • Specific examples of the Bifidobacterium addressntis include ATCC 15703.
  • Specific examples of Bifidobacterium dentium include DSM20436.
  • Specific examples of Bifidobacterium animalis include DSM10140, Bb-12, DN-173010, GCL2505, and CNCM I-3446.
  • Specific examples of the Bifidobacterium pseudolong gum include JCM 5820 and ATCC 25526.
  • Specific examples of the Bifidobacterium thermophyllum include ATCC 25525.
  • strains include, for example, American Type Culture Collection (ATCC, Address: 10801 University Boulevard Manassas, VA20110, United States of America), Belgian Coordinated Collections of Microorganisms (BCCM, Address: Rue de la Science) ), German Collection of Microorganisms and Cell Cultures (DSMZ, Address: Inhoffenstr.7B, D38124 Braunschweig, Germany), Japan Collection of Microorganisms (JCM, Postal Code: 305-0074, Address: 3-Takanodai, Ibaraki, Japan 1-1) Japan Collection of Microorganisms, Bioresource Research Center, Institute of Physical and Chemical Research) or the depositary institution where each strain was deposited.
  • ATCC American Type Culture Collection
  • BCCM Address: Rue de la Science
  • DSMZ German Collection of Microorganisms and Cell Cultures
  • JCM Postal Code: 305-0074, Address: 3-Takanodai, Ibaraki, Japan 1-1) Japan Collection of Microorganism
  • strain specified by the above-exemplified strain name is not limited to the strain itself that has been deposited or registered with a predetermined institution under the strain name (hereinafter, also referred to as "deposited strain” for convenience of explanation).
  • Substantially equivalent strains also referred to as “derivative strains” or “derivative strains” are also included. That is, for example, "Bifidobacterium breve FERM BP-11175" is not limited to the strain itself deposited with the above depositary institution with the deposit number of FERM BP-11175, and includes strains substantially equivalent thereto. Will be done.
  • the "strain substantially equivalent to the above-mentioned deposit strain” belongs to the same species as the above-mentioned deposit strain, and by its use, a desired effect such as a skeletal muscle slowing effect can be obtained, and the 16SrRNA thereof.
  • the base sequence of the gene has preferably 99.86% or more, more preferably 99.93% or more, still more preferably 100% identity with respect to the base sequence of the 16SrRNA gene of the depositary strain, and Preferably, it refers to a strain having the same mycological properties as the deposited strain.
  • a strain substantially equivalent to the above-mentioned deposited strain may be capable of obtaining a desired effect such as a skeletal muscle slowing effect equal to or higher than that of the deposited strain.
  • the “effect equal to or higher than that of the deposited stock” may mean an effect of 80% or more, 90% or more, 95% or more, or 100% or more of the effect obtained by using the deposited stock.
  • a strain substantially equivalent to the above-mentioned deposit strain may be, for example, a derivative strain having the deposit strain as a parent strain.
  • Derivative strains include strains bred from deposit stocks and strains naturally generated from deposit stocks. Examples of the breeding method include modification by a genetic engineering method and modification by mutation treatment.
  • Mutation treatment includes X-ray irradiation, ultraviolet irradiation, and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), ethylmethanesulfonate (EMS), and methylmethanesulfonate (MMS). ) And other treatments with a mutagen.
  • MNNG N-methyl-N'-nitro-N-nitrosoguanidine
  • EMS ethylmethanesulfonate
  • MMS methylmethanesulfonate
  • Bifidobacterium genus bacterium as an active ingredient may mean a bacterial cell of Bifidobacterium genus bacterium.
  • the "product of Bifidobacterium spp.” As an active ingredient may mean a component produced by culturing Bifidobacterium spp.
  • the product produced by the genus Bifidobacterium is not particularly limited as long as it can obtain a desired effect such as a skeletal muscle slowing effect.
  • the products produced by Bifidobacterium are the components that accumulate in the medium during culturing of the bacterium, the components that accumulate in the cells during culturing of the bacterium, and the components that make up the cells of the bacterium. Can be mentioned. Examples of the components constituting the bacterial cells include cell walls and peptidoglycan.
  • the bacterial cells of the genus Bifidobacterium themselves do not have to correspond to "products of the genus Bifidobacterium".
  • the active ingredient a commercially available product may be used, or an appropriately manufactured and obtained product may be used.
  • a commercially available product of the genus Bifidobacterium there is Bifidobacterium Breve FERM BP-11175 manufactured by Morinaga Milk Industry Co., Ltd.
  • the active ingredient can be easily obtained by culturing Bifidobacterium spp.
  • the culture method is not particularly limited as long as Bifidobacterium spp. Can grow.
  • the culturing method for example, a method usually used for culturing Bifidobacterium spp. Can be used as it is or after being appropriately modified.
  • the culture temperature may be, for example, 25 to 50 ° C, preferably 35 to 42 ° C.
  • the culture can be preferably carried out under anaerobic conditions, for example, while aerating an anaerobic gas such as carbon dioxide.
  • the culture can also be carried out under microaerobic conditions such as liquid static culture. Culturing can be carried out, for example, until Bifidobacterium spp. Grow to the desired degree.
  • the medium used for culturing is not particularly limited as long as Bifidobacterium spp. Can grow.
  • a medium usually used for culturing bifidobacteria can be used as it is or after being appropriately modified.
  • the medium may contain, for example, a carbon source, a nitrogen source, an inorganic salt, an organic component, a milk component, or a combination thereof.
  • a carbon source for example, sugars such as glucose, galactose, lactose, arabinose, mannose, sucrose, starch, starch hydrolyzate, and waste sugar honey can be used depending on the assimilation property.
  • ammonium salts such as ammonia, ammonium sulfate, ammonium chloride and ammonium nitrate and nitrates
  • inorganic salt for example, sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, ferrous sulfate and the like
  • organic component for example, peptone, soybean flour, defatted soybean meal, meat extract, yeast extract and the like can be used.
  • milk component for example, milk protein or the like can be used. Milk proteins include casein, whey, and their degradation products.
  • the active ingredient itself or a fraction containing it can be used without particular limitation. That is, as the bifidobacteria bacterium, a bacterial cell of the bifidobacteria genus or a fraction containing the same can be used without particular limitation.
  • the fraction containing a bacterial cell of the genus Bifidobacterium may contain a product of the genus Bifidobacterium in addition to the bacterial cell. Further, as the product produced by the bacterium of the genus Bifidobacterium, a component produced by culturing the bacterium of the genus Bifidobacterium or a fraction containing the same can be used without particular limitation.
  • the fraction containing a component produced by culturing a bifidobacteria bacterium may contain a bifidobacteria bacterium in addition to the component.
  • the active ingredient for example, the culture obtained by culturing may be used as it is, the cells or culture supernatant recovered from the culture may be used, or they may be appropriately treated and used. That is, specific examples of the active ingredient include a culture of a bacterium belonging to the genus Bifidobacterium, a bacterial cell recovered from the same culture, a culture supernatant recovered from the same culture, and a processed product thereof.
  • the treatment is not particularly limited as long as it does not impair a desired effect such as a skeletal muscle slowing effect.
  • Treatments include concentration, dilution, drying, sterilization, crushing and fractionation. These treatments may be carried out alone or in combination as appropriate. All of these treatments can be carried out, for example, by a known method. Drying can be carried out, for example, by freeze-drying.
  • the cells may be crushed and the crushed cells may be used as an active ingredient.
  • the method for crushing the cells include ultrasonic treatment and homogenization treatment.
  • the crushed cells may be used as an active ingredient as it is or after being appropriately subjected to the above-mentioned treatment.
  • a fraction containing the active ingredient may be fractionated from the culture, cells, culture supernatant, or a processed product thereof (for example, crushed cells) as described above and used as the active ingredient. .. Specifically, for example, fractions such as a supernatant fraction and a precipitate fraction may be fractionated from the crushed cell product and used as an active ingredient. Further, for example, the active ingredient may be fractionated (that is, separated and purified) from the culture, cells, culture supernatant, or a processed product thereof (for example, crushed cells) as described above. Separation and purification of the active ingredient may be carried out to the desired extent.
  • Separation and purification of the active ingredient can be carried out, for example, by a known method used for separation and purification of a compound. Such techniques include centrifugation, salting out, solvent extraction, gel filtration, HPLC, membrane separation. For solvent extraction, for example, a polar solvent, a non-polar solvent, or a mixed solvent thereof may be used.
  • a bacterium belonging to the genus Bifidobacterium may be used.
  • the active ingredient may be, in particular, a Bifidobacterium bacterium (specifically, a cell of a Bifidobacterium bacterium or a fraction containing the same).
  • a Bifidobacterium bacterium specifically, a cell of a Bifidobacterium bacterium or a fraction containing the same.
  • Specific examples of the bifidobacteria include cultures of bifidobacteria, bacterial cells recovered from the cultures, and processed products such as dilutions, concentrates, dried products, and sterilized products thereof. Can be mentioned.
  • the bacterial cell may be a live bacterial cell or a dead bacterial cell.
  • the cells may be used, in particular, in a form containing dead cells.
  • the bacterial cell may be composed of, for example, a dead bacterial cell, or may be a mixture of a dead bacterial cell and a live bacterial cell.
  • the ratio of dead cells in the cells may be, for example, 50% or more, 70% or more, 90% or more, 95% or more, 97% or more, or 99% or more based on the number of bacteria.
  • the killed cells can be prepared, for example, by subjecting the live cells to a sterilization treatment.
  • the sterilization treatment includes heat treatment, spray drying method (spray drying method), retort sterilization method, freeze sterilization method, UHT sterilization method, pressure sterilization method, high pressure steam sterilization method, dry heat sterilization method, distribution steam sterilization method, and electromagnetic waves.
  • heat treatment include heating at 70 to 100 ° C. for 10 to 60 minutes.
  • composition of the present invention is a composition containing the above-mentioned active ingredient.
  • the composition of the present invention is a composition containing a bacterium belonging to the genus Bifidobacterium and / or a product of the bacterium.
  • composition of the present invention can be administered to a subject and used. Specifically, the composition of the present invention can be administered to a subject in the manner described in the method of the present invention described later.
  • the composition of the present invention can be used, for example, to obtain the effects illustrated above.
  • the composition of the present invention by administering the composition of the present invention to a subject, the skeletal muscle can be slowed in the subject, that is, the skeletal muscle slowing effect. Is obtained. That is, the composition of the present invention may be a composition for slowing skeletal muscle.
  • the composition of the present invention may be a composition for promoting AMPK activation.
  • One aspect of the AMPK activation promoting composition may be, for example, a skeletal muscle slowing composition.
  • the expression of PGC-1 ⁇ may be promoted in the subject, that is, the expression of PGC-1 ⁇ .
  • a promoting effect may be obtained.
  • the composition of the present invention may be a composition for promoting PGC-1 ⁇ expression.
  • One aspect of the composition for promoting the expression of PGC-1 ⁇ may be, for example, a composition for slowing skeletal muscle.
  • the composition of the present invention may be a composition for improving endurance. Further, the composition of the present invention may be a composition for improving muscle strength. In addition, the composition of the present invention may be a composition for preventing, ameliorating, and / or treating symptoms associated with muscular atrophy and / or muscle weakness. Each of these compositions may be, for example, one aspect of a composition for slowing skeletal muscle.
  • composition of the present invention For the subject to whom the composition of the present invention is administered, the description of the subject to which the active ingredient is administered in the "method of the present invention" can be applied mutatis mutandis.
  • the composition of the present invention may be, for example, a food or drink composition, a pharmaceutical composition, or a feed composition.
  • the composition of the present invention may be, in particular, a food or drink composition or a pharmaceutical composition. That is, the present invention is, for example, a food or drink composition for the above-exemplified use such as for skeletal muscle slowing, a pharmaceutical composition for the above-exemplified use for skeletal muscle slowing, or for skeletal muscle slowing. Etc. may be provided for feed compositions for the above-exemplified uses.
  • composition of the present invention which is a food or drink composition, a pharmaceutical composition, or a feed composition
  • composition of the present invention may consist of an active ingredient or may contain an ingredient other than the active ingredient.
  • Ingredients other than the active ingredient are not particularly limited as long as they do not impair desired effects such as skeletal muscle slowing effect.
  • an acceptable one can be used depending on the usage mode of the composition of the present invention.
  • an ingredient used by blending in foods and drinks, pharmaceuticals, or feeds can be used.
  • Specific examples of the ingredients other than the active ingredient include ingredients such as those exemplified for the food and drink composition, the pharmaceutical composition, and the feed composition described later.
  • the ingredient other than the active ingredient one kind of ingredient may be used, or two or more kinds of ingredients may be used in combination.
  • components other than the active ingredient include components having a muscle-building effect and components that assist the muscle-building effect.
  • components include proteins such as whey protein, casein protein, soybean protein, and pea protein (pea protein), mixtures thereof, and degradation products thereof; such as leucine, valine, isoleucine, and glutamine.
  • Amino acids vitamins such as vitamin B6 and vitamin C; creatin; citric acid; fish oil.
  • the content and content ratio of each component (that is, the active ingredient and optionally other components) in the composition of the present invention are not particularly limited as long as a desired effect such as a skeletal muscle slowing effect can be obtained.
  • the content and content ratio of each component in the composition of the present invention are the type of active ingredient, the type of other components, the type of composition, the shape (dosage form), and the usage, the type of administration target, the age, and It can be set as appropriate according to various conditions such as health condition.
  • the content of the active ingredient in the composition of the present invention is, for example, 1 ⁇ 10 4 cells / g or more, 1 ⁇ 10 5 cells / g or more, and 1 ⁇ 10 6 cells / g in terms of the content of bacterial cells. It may be 1 ⁇ 10 7 cells / g or more, or 1 ⁇ 10 8 cells / g or more, and 10 13 cells / g or less, 10 12 cells / g or less, or 1 ⁇ 10 11 cells / g or less. It may be in the range of combinations thereof.
  • the content of the active ingredient in the composition of the present invention is, for example, 1 ⁇ 10 4 cells / mL or more, 1 ⁇ 10 5 cells / mL or more, and 1 ⁇ 10 6 cells in terms of the content of cells. / ML or more, 1 ⁇ 10 7 cells / mL or more, or 1 ⁇ 10 8 cells / mL or more, 10 13 cells / mL or less, 10 12 cells / mL or less, or 1 ⁇ 10 11 cells / mL It may be the following, or it may be a range of combinations thereof.
  • the content of the active ingredient in the composition of the present invention is, for example, 1 ⁇ 10 6 to 1 ⁇ 10 12 cells / g or 1 ⁇ 10 6 to 1 ⁇ in terms of the content of bacterial cells.
  • 10 12 cells / mL preferably 1 ⁇ 10 7 to 1 ⁇ 10 11 cells / g or 1 ⁇ 10 7 to 1 ⁇ 10 11 cells / mL, more preferably 1 ⁇ 10 8 to 1 ⁇ 10 10 cells / g or It may be 1 ⁇ 10 8 to 1 ⁇ 10 10 cells / mL.
  • “cells” may be read as “cfu”.
  • “Cfu” represents a colony forming unit.
  • the content of the active ingredient in the composition of the present invention is, for example, 0.01% (w / w) or more, 0.1% (w / w) or more, 1 in terms of the content of the original culture. % (W / w) or more, 5% (w / w) or more, or 10% (w / w) or more, 10,000% (w / w) or less, 5000% (w / w) or less, 1000% (w / w) or less, 500% (w / w) or less, 300% (w / w) or less, 200% (w / w) or less, 150% (w / w) or less, 100% (w / w / w) or less, 70% (w / w) or less, 50% (w / w) or less, 30% (w / w) or less, 10% (w / w) or less, 5% (w / w) or less, or It may be 1% (w
  • the content of the active ingredient in the composition of the present invention is, for example, 1 to 10000% (w / w), preferably 10 to 1000% (w) in terms of the content of the original culture. / W) may be used. That is, for example, the composition of the present invention may contain a culture of a bacterium belonging to the genus Bifidobacterium or a product thereof (culture supernatant, etc.) in a content as exemplified above.
  • the "original culture” refers to a culture in which a concentration change such as concentration or dilution does not occur after the culture, and specifically, the culture immediately after the culture (that is, the culture of Bifidobacterium spp. ) May be.
  • Such cultures are, for example, 1 ⁇ 10 7 to 1 ⁇ 10 11 cfu / mL, preferably 1 ⁇ 10 8 to 1 ⁇ 10 10 cfu for Bifidobacterium using the medium as exemplified above.
  • the culture may be cultured so as to be / mL.
  • a content exceeding 100% (w / w) means that the culture or a derivative thereof (culture supernatant, etc.) is concentrated and contained in the composition of the present invention. That is, for example, a content of 1000% (w / w) means that the culture or a derivative thereof (culture supernatant, etc.) is concentrated 10 times and contained in the composition of the present invention.
  • the content of the active ingredient in the composition of the present invention can be set so that, for example, the dose of the active ingredient described later can be obtained. That is, the dose of the composition of the present invention is, for example, 1 ⁇ 10 6 to 1 ⁇ 10 12 cells / kg body weight / day, preferably 1 ⁇ 10 in terms of the dose of the active ingredient (cell-based). It may be 7 to 1 ⁇ 10 11 cells / kg body weight / day, more preferably 1 ⁇ 10 8 to 1 ⁇ 10 10 cells / kg body weight / day.
  • the dose of the composition of the present invention is, for example, 1 ⁇ 10 7 to 1 ⁇ 10 14 cells / day / individual, preferably 1 ⁇ 10 8 in terms of the dose of the active ingredient (cell-based). It may be ⁇ 1 ⁇ 10 13 cells / day / individual, more preferably 1 ⁇ 10 9 to 1 ⁇ 10 12 cells / day / individual. When the active ingredient is a viable cell, "cells" may be read as "cfu”.
  • the dose of the composition of the present invention is, for example, 0.01 to 100 mL / kg body weight / day, preferably 0.1 to 10 mL / day in terms of the dose of the active ingredient (based on the original culture). It may be kg body weight / day.
  • the shape of the composition of the present invention is not particularly limited. As the shape of the composition of the present invention, an acceptable one can be adopted depending on the usage mode of the composition of the present invention. Specific examples of the shape of the composition of the present invention include shapes such as those exemplified for the food and drink composition, the pharmaceutical composition, and the feed composition described later.
  • the food and drink composition of the present invention is not particularly limited as long as it contains an active ingredient.
  • the food and drink composition of the present invention may be provided in any form such as liquid, paste-like, gel-like solid, and powder.
  • the food and drink composition may be, for example, the food and drink itself, or may be a material used for manufacturing the food and drink.
  • Such ingredients include seasonings, food additives and other food and beverage ingredients.
  • Specific examples of food and drink compositions include wheat flour products, instant foods, processed agricultural products, processed marine products, processed livestock products, dairy products (fermented milk, cheese, prepared milk powder for childcare, etc.), fats and oils, and basic seasonings. , Complex seasonings, frozen foods, confectionery, beverages, and other commercially available foods and drinks.
  • food and drink compositions specifically, health foods, functional foods, enteric nutritional foods, special purpose foods, health functional foods (foods for specified health use, nutritional functional foods, foods with functional claims, etc.), Food supplements and non-medicinal products are also included.
  • the food and drink composition may be, for example, a supplement, and specifically, a tablet-shaped supplement.
  • the food and drink composition of the present invention can be produced by combining the active ingredient with other ingredients.
  • the operation of combining the active ingredient with other ingredients is also called "addition of the active ingredient".
  • the method for producing the food and drink composition of the present invention is not particularly limited.
  • the food and drink composition of the present invention can be produced, for example, by the same method as for ordinary food and drink, using the same raw materials as for ordinary food and drink, except that the active ingredient is added.
  • the addition of the active ingredient may be carried out at any stage of the manufacturing process of the food or drink composition.
  • the addition of the active ingredient may be carried out, for example, during or after the production of the food or drink composition. That is, for example, the food or drink composition of the present invention may be obtained by adding an active ingredient to a food or drink prepared in advance.
  • a food or drink composition may be produced through a fermentation step using the added active ingredient (particularly Bifidobacterium spp.).
  • the food and drink composition produced through the fermentation step include fermented products such as lactic acid bacteria beverages and fermented milk. That is, the active ingredient (particularly Bifidobacterium spp.) May be used, for example, as a starter for the production of fermented products. The active ingredient can also be added later to the fermented product produced.
  • another food or drink composition can be produced by using the food or drink composition of the present invention. That is, for example, when the food and drink composition of the present invention is provided as a material used for producing food and drink such as seasonings, food additives, and other food and drink raw materials, the present invention is used as a raw material for food and drink. Another food and drink composition can be produced by adding the food and drink composition. Another food or drink composition produced in this manner is also an aspect of the food or drink composition of the present invention. Regarding the addition of the food and drink composition of the present invention, the description regarding the addition of the active ingredient in the production of the food and drink composition can be applied mutatis mutandis.
  • the food and drink composition of the present invention is used for slowing skeletal muscle, promoting AMPK activation, promoting PGC-1 ⁇ expression, preventing, ameliorating, and / or treating symptoms associated with muscular atrophy and / or muscle weakness. It is possible to provide and sell as food and drink with indications such as (including health use).
  • the food and drink composition of the present invention is intended to be ingested by "athletes”, “those who want to gain muscle”, “those who are concerned about muscle weakness due to lack of exercise", and "age-related decline”. It is possible to provide and sell by displaying "people who are concerned about muscles", “people who are concerned about muscle strength that declines with age”, “people who want to improve flail", etc.
  • Display includes all actions for informing the consumer of the use, and if the expression is such that the use can be recalled or analogized, the purpose of the display, the content of the display, and the display are displayed. Regardless of the object, medium, etc., all fall under “display”. In particular, the display is preferably performed in such a way that the consumer can directly recognize the application.
  • the product or the packaging of the product according to the food or drink composition of the present invention in which the above-mentioned use is described, is transferred, transferred, displayed for transfer or delivery, and imported.
  • the above-mentioned uses are described in the acts, advertisements, price lists or transaction documents related to the products, displayed or distributed, or the above-mentioned uses are described in the information containing these and provided by an electromagnetic (Internet, etc.) method. Acts, etc. can be mentioned.
  • Examples of the display include labeling on packaging, containers, catalogs, pamphlets, promotional materials such as POPs at sales sites, and other documents.
  • a display approved by the government or the like for example, a display obtained based on various systems established by the government and performed in a manner based on such approval
  • labeling health foods, functional foods, enteral nutrition foods, special purpose foods, health functional foods (for example, specified health foods, nutritional functional foods, functional foods), nutritional supplements, pharmaceutical departments Labeling as a foreign product can be mentioned.
  • Examples of the labeling approved by the government, etc. include the labeling approved by the Consumer Affairs Agency. Labels approved by the Consumer Affairs Agency include those approved by foods for specified health uses and similar systems.
  • the labeling approved by the Consumer Affairs Agency includes labeling as a food for specified health use, labeling as a food for specified health use with conditions, labeling indicating that it affects the structure and function of the body, and reduction of disease risk. Labeling, labeling of functionality based on scientific grounds, etc., and more specifically, Cabinet Office Ordinance on permission of labeling for special purposes stipulated in the Health Promotion Law (March 31, 2001) Labeling as food for specified health use (particularly labeling for health use) and similar labeling specified in Cabinet Office Ordinance No. 57) of Japan can be mentioned.
  • the content of the active ingredient in the food and drink composition of the present invention may be, for example, in the above range. That is, the content of the active ingredient in the food or drink composition of the present invention is specifically, for example, 1 ⁇ 10 6 to 1 ⁇ 10 12 cells / g or 1 ⁇ 10 in terms of the content of bacterial cells. 6 to 1 ⁇ 10 12 cells / mL, preferably 1 ⁇ 10 7 to 1 ⁇ 10 11 cells / g or 1 ⁇ 10 7 to 1 ⁇ 10 11 cells / mL, more preferably 1 ⁇ 10 8 to 1 ⁇ 10 10 It may be cells / g or 1 ⁇ 10 8 to 1 ⁇ 10 10 cells / mL.
  • the content of the active ingredient in the food and drink composition of the present invention is specifically, for example, 1 to 10000% (w / w), preferably 10 to 1 in terms of the content of the original culture. It may be 1000% (w / w).
  • the dose of the food or drink composition of the present invention may be, for example, in the above range. That is, the dose of the food or drink composition of the present invention is specifically converted into, for example, the dose of the active ingredient (based on the amount of bacterial cells), and is 1 ⁇ 10 6 to 1 ⁇ 10 12. It may be cells / kg body weight / day, preferably 1 ⁇ 10 7 to 1 ⁇ 10 11 cells / kg body weight / day, more preferably 1 ⁇ 10 8 to 1 ⁇ 10 10 cells / kg body weight / day. In addition, the dose of the food or drink composition of the present invention is specifically converted into, for example, the dose of the active ingredient (based on the amount of bacterial cells) and 1 ⁇ 10 7 to 1 ⁇ 10 14.
  • the dose of the food or drink composition of the present invention is, specifically, 0.01 to 100 mL / kg in terms of the dose of the active ingredient (based on the amount of the original culture). It may be body weight / day, preferably 0.1-10 mL / kg body weight / day.
  • the pharmaceutical composition of the present invention is not particularly limited as long as it contains an active ingredient.
  • the active ingredient may be used as it is, or the active ingredient may be appropriately formulated and used.
  • the dosage form of the pharmaceutical composition of the present invention is not particularly limited.
  • the dosage form of the pharmaceutical composition of the present invention can be appropriately selected depending on various conditions such as the administration method.
  • the pharmaceutical composition of the present invention may be for oral administration or parenteral administration.
  • the pharmaceutical composition of the present invention may be, in particular, for oral administration.
  • specific dosage forms include solid preparations such as powders, granules, tablets and capsules, and liquid preparations such as solutions, syrups, suspensions and emulsions.
  • specific dosage forms include suppositories and ointments.
  • Physiologically acceptable additives can be used in the formulation. Additives include various organic and inorganic components. Specific examples of the additive include excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents, diluents, surfactants, and solvents. These additives can be appropriately selected depending on various conditions such as dosage form.
  • excipients include sugar derivatives such as lactose, sucrose, glucose, mannit, and sorbit; starch derivatives such as corn starch, horse bell starch, ⁇ -starch, dextrin, and carboxymethyl starch; crystalline cellulose, hydroxypropyl cellulose, etc.
  • Cellulose derivatives such as hydroxypropylmethyl cellulose, carboxymethyl cellulose, carboxymethyl cellulose calcium; gum arabic; dextran; purulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, magnesium aluminometasilicate; phosphate derivatives such as calcium phosphate; carbonic acid Carbonated carbonate derivatives such as calcium; sulfate derivatives such as calcium sulfate can be mentioned.
  • binder examples include gelatin; polyvinylpyrrolidone; macrogol, etc., in addition to the above-mentioned excipients.
  • disintegrant examples include, in addition to the above-mentioned excipients, chemically modified starch or cellulose derivatives such as croscarmellose sodium, carboxymethyl starch sodium, and crosslinked polyvinylpyrrolidone.
  • lubricant examples include talc; stearic acid; metal stearic acid salts such as calcium stearate and magnesium stearate; colloidal silica; waxes such as pea gum and gay wax; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid.
  • Sodium carboxylic acid salts such as sodium benzoate; sulfates such as sodium sulfate; leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acids such as silicic acid anhydride and silicate hydrate; starch derivatives and the like. Be done.
  • the stabilizer examples include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol and phenylethyl alcohol; benzalkonium chloride; acetic acid anhydride; and sorbic acid.
  • flavoring and flavoring agent examples include sweeteners, acidulants, and flavors.
  • the content of the active ingredient in the pharmaceutical composition of the present invention may be, for example, in the above range. That is, specifically, the content of the active ingredient in the pharmaceutical composition of the present invention is, for example, 1 ⁇ 10 6 to 1 ⁇ 10 12 cells / g or 1 ⁇ 10 6 in terms of the content of bacterial cells. ⁇ 1 ⁇ 10 12 cells / mL, preferably 1 ⁇ 10 7 to 1 ⁇ 10 11 cells / g or 1 ⁇ 10 7 to 1 ⁇ 10 11 cells / mL, more preferably 1 ⁇ 10 8 to 1 ⁇ 10 10 cells. It may be / g or 1 ⁇ 10 8 to 1 ⁇ 10 10 cells / mL.
  • the active ingredient When the active ingredient is a viable cell, “cells” may be read as “cfu”. Further, specifically, the content of the active ingredient in the pharmaceutical composition of the present invention is, for example, 1 to 10000% (w / w), preferably 10 to 1000, in terms of the content of the original culture. It may be% (w / w).
  • the dose of the pharmaceutical composition of the present invention may be, for example, in the above range. That is, specifically, the dose of the pharmaceutical composition of the present invention is converted into, for example, the dose of the active ingredient (based on the amount of bacterial cells) and 1 ⁇ 10 6 to 1 ⁇ 10 12 cells. It may be / kg body weight / day, preferably 1 ⁇ 10 7 to 1 ⁇ 10 11 cells / kg body weight / day, more preferably 1 ⁇ 10 8 to 1 ⁇ 10 10 cells / kg body weight / day. Further, the dose of the pharmaceutical composition of the present invention is specifically converted into, for example, the dose of the active ingredient (based on the amount of bacterial cells) and 1 ⁇ 10 7 to 1 ⁇ 10 14 cells.
  • the dose of the pharmaceutical composition of the present invention is, for example, 0.01 to 100 mL / kg body weight in terms of the dose of the active ingredient (based on the amount of the original culture). It may be / day, preferably 0.1-10 mL / kg body weight / day.
  • the feed composition of the present invention is not particularly limited as long as it contains an active ingredient.
  • Examples of the feed composition include pet food and livestock feed.
  • the feed of the present invention may be provided in any form such as powder, granule, crumble, pellet, cube, paste and liquid.
  • the feed composition of the present invention can be produced by combining the active ingredient with other ingredients.
  • the operation of combining the active ingredient with other ingredients is also called "addition of the active ingredient".
  • the method for producing the feed composition of the present invention is not particularly limited.
  • the feed composition of the present invention can be produced, for example, by the same method as a normal feed using the same raw materials as a normal feed, except that an active ingredient is added.
  • the addition of the active ingredient may be carried out at any stage of the production process of the feed composition.
  • the addition of the active ingredient may be carried out, for example, during or after the production of the feed composition. That is, for example, the feed composition of the present invention may be obtained by adding an active ingredient to a feed prepared in advance.
  • the feed composition may be produced through a fermentation step using the added active ingredient (particularly Bifidobacterium spp.). Silage is mentioned as a feed composition produced through a fermentation step.
  • Method of the present invention is a method including a step of administering the above-mentioned active ingredient to a subject. This process is also referred to as "administration process”.
  • the method of the present invention is a method including a step of administering a bacterium belonging to the genus Bifidobacterium and / or a product of the bacterium to a subject.
  • the method of the present invention specifically, by administering the active ingredient to a subject, skeletal muscle can be slowed in the subject, that is, a skeletal muscle slowing effect can be obtained. That is, the method of the present invention may be a method of slowing down skeletal muscle.
  • the activation of AMPK may be promoted in the subject, that is, the AMPK activation promoting effect may be obtained. That is, the method of the present invention may be a method of promoting activation of AMPK.
  • One aspect of the method of promoting the activation of AMPK may be, for example, a method of slowing down skeletal muscle.
  • the method of the present invention specifically, by administering the active ingredient to a subject, the expression of PGC-1 ⁇ may be promoted in the subject, that is, the PGC-1 ⁇ expression promoting effect may be obtained. That is, the method of the present invention may be a method of promoting the expression of PGC-1 ⁇ .
  • One aspect of the method of promoting the expression of PGC-1 ⁇ may be, for example, a method of slowing down skeletal muscle.
  • the method of the present invention may be a method for improving endurance. Further, the method of the present invention may be a method for improving muscle strength. The method of the present invention may also be a method of preventing, ameliorating, and / or treating symptoms associated with muscular atrophy and / or weakness. Each of these methods may be, for example, one aspect of a method of slowing skeletal muscle.
  • administering the active ingredient to the subject may be synonymous with “making the subject ingest the active ingredient”.
  • Ingestion may be voluntary (free intake) or compulsory (forced intake). That is, specifically, the administration step may be, for example, a step of blending the active ingredient with food or drink or feed and supplying it to the subject, thereby allowing the subject to freely ingest the active ingredient.
  • the administration may be oral administration or parenteral administration.
  • the administration may usually be oral administration.
  • Parenteral administration includes tube administration and rectal administration.
  • the administration mode of the active ingredient (for example, administration target, administration time, administration period, number of administrations, dose, and other conditions relating to administration) is not particularly limited as long as a desired effect such as a skeletal muscle slowing effect can be obtained. ..
  • the administration mode of the active ingredient can be appropriately set according to various conditions such as the type of the active ingredient, the type of the subject to be administered, the age, and the health condition.
  • the subject to which the active ingredient is administered is not particularly limited as long as a desired effect such as a skeletal muscle slowing effect can be obtained.
  • Mammals are examples of subjects to which the active ingredient is administered. Mammals include humans and pets. Examples of pets include dogs and cats. Mammals include, in particular, humans. The mammal may be male or female.
  • the human conditions under which the active ingredient is administered are not particularly limited.
  • the active ingredient may be administered to any human who desires a desired effect, such as a skeletal muscle slowing effect.
  • the human may be, for example, an infant, a child, an adult, a middle-aged person, an elderly person, or any other age group.
  • the human may be, for example, a healthy person or an unhealthy person. Unhealthy individuals include those who exhibit symptoms associated with muscle atrophy and / or weakness. Humans may or may not be those who perform physical exercises other than daily activities, such as athletes and sports enthusiasts.
  • the dose of the active ingredient is, for example, 1 ⁇ 10 6 cells / kg body weight / day or more, 1 ⁇ 10 7 cells / kg body weight / day or more, or 1 ⁇ 10 8 cells / day in terms of the dose of cells. It may be kg body weight / day or more, 1 ⁇ 10 12 cells / kg body weight / day or less, 1 ⁇ 10 11 cells / kg body weight / day or less, or 1 ⁇ 10 10 cells / kg body weight / day or less. It may be a range of combinations thereof.
  • the dose of the active ingredient is, for example, 1 ⁇ 10 6 to 1 ⁇ 10 12 cells / kg body weight / day, preferably 1 ⁇ 10 7 to 1 ⁇ 10 in terms of the dose of bacterial cells. It may be 11 cells / kg body weight / day, more preferably 1 ⁇ 10 8 to 1 ⁇ 10 10 cells / kg body weight / day.
  • “cells” may be read as "cfu”.
  • the dose of the active ingredient is, for example, 1 ⁇ 10 7 cells / day / individual or more, 1 ⁇ 10 8 cells / day / individual or more, or 1 ⁇ 10 9 cells / day / day in terms of the dose of cells. It may be more than an individual, 1 ⁇ 10 14 cells / day / individual or less, 1 ⁇ 10 13 cells / day / individual or less, or 1 ⁇ 10 12 cells / day / individual or less, and a combination thereof. It may be in the range of. Specifically, the dose of the active ingredient is, for example, 1 ⁇ 10 7 to 1 ⁇ 10 14 cells / day / individual, preferably 1 ⁇ 10 8 to 1 ⁇ 10 13 in terms of the dose of bacterial cells. It may be cells / day / individual, more preferably 1 ⁇ 10 9 to 1 ⁇ 10 12 cells / day / individual. When the active ingredient is a viable cell, "cells" may be read as "cfu".
  • the dose of the active ingredient may be, for example, 0.01 mL / kg bw / day or more, or 0.1 mL / kg bw / day or more, or 100 mL / kg in terms of the dose of the original culture. It may be body weight / day or less, or 10 mL / kg body weight / day or less, or a combination thereof. Specifically, the dose of the active ingredient is, for example, 0.01 to 100 mL / kg body weight / day, preferably 0.1 to 10 mL / kg body weight / day in terms of the dose of the original culture. There may be. That is, for example, a culture of a bacterium belonging to the genus Bifidobacterium or a product thereof (culture supernatant, etc.) may be administered at a dose as exemplified above.
  • the administration period of the active ingredient may be, for example, 4 weeks or longer, preferably 8 weeks or longer, and more preferably 12 weeks or longer.
  • the active ingredient may be administered once per day, for example, or may be administered in multiple divided doses per day.
  • the active ingredient may be administered, for example, daily or once every few days.
  • the active ingredient may be administered daily, in particular.
  • the dose of the active ingredient at each dose may or may not be constant.
  • the active ingredient may be administered to the subject as it is, or may be prepared as a composition such as a food or drink composition, a pharmaceutical composition, a feed composition, etc. containing the active ingredient and administered to the subject.
  • the description of the composition of the present invention can be applied mutatis mutandis to the composition containing the active ingredient.
  • the active ingredient may be administered alone or in combination with other ingredients.
  • other components include pharmaceuticals and pharmaceutical compositions, foods and drinks and food and drink compositions, feeds and feed compositions, and components contained therein. These other components may or may not be used for the above-exemplified uses such as for slowing skeletal muscle.
  • the active ingredient can also be administered to a subject using, for example, the composition of the present invention (that is, by administering the composition of the present invention). That is, one aspect of the method of the present invention may be a method including a step of administering the composition of the present invention to a subject. That is, the "administration of the active ingredient” also includes the administration of the composition of the present invention.
  • the administration mode of the composition of the present invention (for example, administration subject, administration time, administration period, number of administrations, dose, and other conditions relating to administration) is as long as a desired effect such as a skeletal muscle slowing effect can be obtained. There are no particular restrictions.
  • the administration mode of the composition of the present invention includes the type and content of the active ingredient, the type and content of other ingredients, the type and shape (dosage form) of the composition, the type of administration target, the age, and the health condition. It can be set as appropriate according to various conditions.
  • the description regarding the administration mode of the active ingredient as described above can also be applied mutatis mutandis when the composition of the present invention is administered to a subject. That is, the composition of the present invention can be administered to, for example, a subject as exemplified above.
  • the dose of the composition of the present invention can be set so as to obtain, for example, the dose of the active ingredient as exemplified above.
  • the composition of the present invention may be administered alone, and may be administered with other components such as pharmaceuticals, pharmaceutical compositions, foods and drinks, food and drink compositions, feeds and feed compositions, and components contained therein. It may be administered in combination.
  • Example 10 Preparation of administration product> The freeze-dried powder of Bifidobacterium breve FERM BP-11 175 with saline and diluted to a concentration of 5 ⁇ 10 8 cfu / mL, to obtain viable cells administered products. A viable cell-administered product of Bifidobacterium Breve FERM BP-11175 was heated at 90 ° C. for 30 minutes to obtain a heat-sterilized product.
  • the experimental animals were allowed to freely ingest Lab MR stock feed (manufactured by Northan Industries) and tap water after purchase at the age of 7 weeks. After acclimatization for 1 week, the experimental animals were controlled (5 mL / kg body weight / day of physiological saline) and live Bifidobacterium cells (2 mL / day / day of live cells) (bacteria).
  • Bifidobacterium heat sterilizer administration group (2 mL / day / animal in heat sterilizer / animal (1 ⁇ 10 9 cells / animal)
  • oral administration of the administered product using an oral sonde was continued once a day for 4 weeks in each mouse.
  • the experimental animals were treated under sevoflurane anesthesia to remove the soleus and gastrocnemius muscles.
  • the gastrocnemius muscle was used to measure the proportion of slow muscle fibers by the following procedure. That is, the excised gastrocnemius muscle was sliced, and slow muscle fibers were stained by immunohistochemical staining using Anti-SERCA2 ATPase antibody (Abcam). For each group, three visual fields of the central part of the medial gastrocnemius muscle were observed under a microscope, and the staining rate (slow muscle fiber ratio) was measured for a total of 450 fibers.
  • Figure 1 shows the proportion of slow muscle fibers in each group.
  • the slow muscle fiber staining images of each group are shown in FIGS. 2 to 4.
  • dark areas are slow muscle fibers and bright areas are fast muscle fibers.
  • Administration of Bifidobacterium breve FERM BP-11175 increased the proportion of slow muscle fibers compared to the control.
  • a marked increase in the proportion of slow muscle fibers was confirmed in the heat-sterilized body-administered group than in the live-cell-administered group.
  • the slow muscle fiber ratio increased by 3.80% in the live cell administration group
  • the slow muscle fiber ratio increased by 12.70% in the heat sterilized body administration group.
  • the soleus muscle is a signal pathway involved in slowing skeletal muscle (Zhang et al., Int J Biol Sci, 13 (9), 1152-1162 (2017); Suwa et al., J. Appl. It was used for the evaluation of Physiol, 95, 960-968 (2003).
  • AMPK activated AMP-activated protein kinase
  • PPC-1 ⁇ peroxysome growth factor activating receptor ⁇ -conjugating factor-1 ⁇
  • RNA was extracted from a part of the excised soleus muscle using RNeasy Mini Kit (Qiagen), and a complementary DNA (cDNA) library was prepared by PrimeScript RT Reagent Kit (Takara Bio Inc.).
  • cDNA complementary DNA
  • SYBR Premix Ex Taq2 (Takara Bio Inc.)
  • PCR was performed to quantify the expression level of PGC-1 ⁇ at the mRNA level. At this time, the expression level of PGC-1 ⁇ was standardized based on the expression level of GAPDH and calculated as a relative value with respect to the control.
  • the expression level of PGC-1 ⁇ is shown in FIG. Administration of Bifidobacterium breve FERM BP-11175 increased the expression level of PGC-1 ⁇ compared with the control. In particular, a marked increase in the expression level of PGC-1 ⁇ was confirmed in the heat-sterilized body-administered group than in the live-cell-administered group. Specifically, the expression level of PGC-1 ⁇ increased 1.31 times in the viable cell administration group and 1.90 times in the heat sterilized body as compared with the control. ..
  • Protein was extracted from a part of the excised soleus muscle using RIPA buffer (CST), and the amount of phosphorylated AMPK (active AMPK) was quantified by Western blotting. At this time, the phosphorylated AMPK amount was standardized based on the total AMPK amount and calculated as a relative value with respect to the control.
  • CST RIPA buffer
  • the amount of phosphorylated AMPK is shown in FIG.
  • Administration of Bifidobacterium Breve FERM BP-11175 increased the amount of active AMPK compared to the control. Specifically, the amount of active AMPK increased 1.42 times in the viable cell administration group and 1.48 times the amount of active AMPK in the heat sterilized body administration group as compared with the control.
  • compositions that can be used for a specific purpose, such as a composition for slowing skeletal muscle.

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Abstract

L'invention concerne une composition, par exemple une composition destinée à convertir un muscle squelettique en muscle lent, pouvant être utilisée à des fins spécifiques. L'invention concerne plus particulièrement une composition contenant comme principe actif des bactéries Bifidobacterium.
PCT/JP2020/014079 2019-03-29 2020-03-27 Composition destinée à convertir un muscle squelettique en muscle lent WO2020203792A1 (fr)

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JP7475155B2 (ja) 2020-02-14 2024-04-26 森永乳業株式会社 持久力向上用組成物

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115105534A (zh) * 2022-07-06 2022-09-27 中国农业大学 动物双歧杆菌a6在治疗肌少症中的应用
CN115105534B (zh) * 2022-07-06 2024-01-26 中国农业大学 动物双歧杆菌a6在治疗肌少症中的应用

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