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WO2023277636A1 - Composition anti-âge contenant une souche de lactobacillus sp. et une substance à base de plantes médicinales pour thérapie combinée - Google Patents

Composition anti-âge contenant une souche de lactobacillus sp. et une substance à base de plantes médicinales pour thérapie combinée Download PDF

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Publication number
WO2023277636A1
WO2023277636A1 PCT/KR2022/009477 KR2022009477W WO2023277636A1 WO 2023277636 A1 WO2023277636 A1 WO 2023277636A1 KR 2022009477 W KR2022009477 W KR 2022009477W WO 2023277636 A1 WO2023277636 A1 WO 2023277636A1
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Prior art keywords
lactobacillus
active material
active
strain
muscle
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English (en)
Korean (ko)
Inventor
양보기
장명호
이한성
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GI Biome Inc
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GI Biome Inc
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Priority claimed from KR1020220080567A external-priority patent/KR20230005057A/ko
Publication of WO2023277636A1 publication Critical patent/WO2023277636A1/fr
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A61K31/525—Isoalloxazines, e.g. riboflavins, vitamin B2
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/733—Fructosans, e.g. inulin
    • 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
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/07—Basidiomycota, e.g. Cryptococcus
    • A61K36/074—Ganoderma
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
    • A61K36/258—Panax (ginseng)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/894—Dioscoreaceae (Yam family)
    • A61K36/8945—Dioscorea, e.g. yam, Chinese yam or water yam
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
    • A61K36/8994—Coix (Job's tears)
    • 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
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • It relates to an anti-aging composition in which a strain of the genus Lactobacillus and herbal medicine are co-administered.
  • Korean Registered Patent Nos. 10-2223248 and 10-0950164 disclose pharmaceutical compositions for the prevention and treatment of sarcopenia or degenerative brain diseases containing herbal medicines as active ingredients, including herbal preparations for the treatment of aging-related diseases. Plans for its use are being actively proposed. However, the effect has not reached a level sufficient to be commercially successful. There is a need for continuous research on new strains and therapeutic agents with excellent anti-aging effects.
  • Another aspect is an anti-aging or antioxidant composition
  • a first active material containing a Lactobacillus sp. strain or a culture thereof as an active ingredient and a second active material containing oriental medicines in combination. is to provide
  • compositions for preventing, improving or treating aging-related diseases comprising a first active material including a Lactobacillus sp. strain or a culture thereof and a second active material including oriental medicine as an active ingredient. is to provide
  • Another aspect is the prevention of aging-related diseases in which a first active material containing a Lactobacillus sp. strain or a culture thereof as an active ingredient and a second active material containing herbal medicines are administered in combination. , to provide a composition for improvement or treatment.
  • Lactobacillus ( Lactobacillus sp. )
  • a first active substance containing a strain or a culture thereof and a second active substance containing herbal medicine health for prevention or improvement of muscle-related diseases or obesity, including as an active ingredient a first active substance containing a strain or a culture thereof and a second active substance containing herbal medicine health to provide functional foods.
  • Another aspect is a muscle-related disease or obesity in which a first active material containing a Lactobacillus sp. strain or a culture thereof as an active ingredient and a second active material containing herbal medicines are administered in combination.
  • a first active material containing a Lactobacillus sp. strain or a culture thereof as an active ingredient and a second active material containing herbal medicines are administered in combination.
  • Another aspect is a muscle-related disease or obesity in which a first active material containing a Lactobacillus sp. strain or a culture thereof as an active ingredient and a second active material containing herbal medicines are administered in combination.
  • compositions comprising, as active ingredients, a first active material including a Lactobacillus sp. strain or a culture thereof and a second active material including herbal medicine for use in the preparation of an anti-aging composition; Or to provide a use of a composition comprising the first active material as an active ingredient and the second active material being co-administered.
  • Another aspect includes administering an effective amount of a first active material comprising a Lactobacillus sp. strain or a culture thereof and a second active material comprising an effective amount of herbal medicine to a subject in need thereof It is to provide a method for preventing or treating muscle-related diseases.
  • Lactobacillus for the production of health functional food for preventing or improving muscle-related diseases ( Lactobacillus sp. )
  • a first active material comprising a strain or a culture thereof and a second active material comprising a herbal medicine
  • a composition containing as an active ingredient Alternatively, it is to provide a use of a composition comprising the first active material as an active ingredient and co-administered with the second active material.
  • Another aspect includes administering an effective amount of a first active material comprising a Lactobacillus sp. strain or a culture thereof and a second active material comprising an effective amount of herbal medicine to a subject in need thereof It is to provide a method for preventing or treating obesity.
  • Another aspect is a first active material comprising a Lactobacillus sp. strain or a culture thereof for the production of health functional food for preventing or improving obesity, and a second active material comprising herbal medicine as an active ingredient.
  • a composition comprising; Alternatively, it is to provide a use of a composition comprising the first active material as an active ingredient and co-administered with the second active material.
  • Another aspect is a first active material comprising a Lactobacillus sp. strain or a culture thereof for the preparation of a pharmaceutical formulation for preventing or treating obesity and a second active material comprising herbal medicine as an active ingredient.
  • Health functional food composition comprising a; Alternatively, it is to provide a use of a composition comprising the first active material as an active ingredient and co-administered with the second active material.
  • One aspect includes a first active material including a Lactobacillus sp. strain or a culture thereof and a second active material including herbal medicine as an active ingredient;
  • a first active material including a Lactobacillus sp. strain or a culture thereof and a second active material including herbal medicine as an active ingredient;
  • an anti-aging composition, an antioxidant composition, or a composition for preventing, improving, or treating aging-related diseases comprising the first active material as an active ingredient and co-administered with the second active material is provided.
  • the first active material is Lactobacillus sakei , Lactobacillus acidophilus , Lactobacillus casei , Lactobacillus gasseri , Lactobacillus paracasei , Lactobacillus plantarum , Lactobacillus ferementum , Lactobacillus rhamnosus , Lactobacillus reuteri , Lactobacillus delbru It may be at least one selected from the group consisting of Lactobacillus delbrueckii , Lactobacillus johnsonii , and Lactobacillus helveticus .
  • the anti-aging may be at least one from the group consisting of anti-aging of skeletal muscle cells, anti-aging of nerve cells, anti-aging of skin cells, anti-aging of immune cells, and anti-aging of the intestinal microbial environment.
  • the pharmaceutical composition for preventing or treating muscle-related diseases may reduce at least one or more from the group consisting of joint stiffness, muscle loss, power loss, speed loss, balance loss, endurance loss, and agility loss. there is.
  • the pharmaceutical composition for preventing or treating muscle-related diseases increases exercise performance, restores muscle coordination, mobility and gait, increases muscle mass and strength, or increases the strength of an elderly adult. It may be to inhibit muscle strength loss.
  • the muscle-related disease is sarcopenia, geriatric sarcopenia, muscular atrophy, muscular dystrophy, disuse muscle atrophy, motor neuron disease , inflammatory myopathy, neuromuscular junction disease, endocrine myopathy, muscle degeneration, muscle stiffness, amyotrophic sclerosis, myasthenia gravis, myositis, muscle calcification, muscle ossification, one or more selected from the group consisting of muscle weakness-related diseases and cachexia it could be
  • the pharmaceutical composition for preventing or treating obesity is from the group consisting of inhibiting fat accumulation in white adipose tissue, reducing metabolic inflammation in adipose tissue, reducing insulin resistance, and improving intestinal microbial composition. At least one selected may be selected.
  • the composition may be administered in combination with the first active material, the second active material, and the third active material simultaneously, sequentially or in reverse order.
  • the composition includes a first oral preparation containing the first active material, a second oral preparation containing a second active material, and a third oral preparation containing a third active material, wherein the The first, second, and third oral preparations may be administered orally.
  • the strain is freeze-dried, and may be contained in an amount of 10 3 to 10 16 CFU/g.
  • the aging-related diseases such as alleviating the symptoms of muscle weakness induced by aging or reducing the change in the intestinal microbial environment induced by aging, It can be usefully used for prevention or treatment.
  • 1 is a graph showing the change in evenness of the flora according to the combined administration of mixed strains and herbal medicines according to one embodiment.
  • Figure 2 is a graph showing the results of intestinal flora principal coordinates analysis (PCoA) according to the combined administration of mixed strains and herbal medicines according to one embodiment.
  • PCoA principal coordinates analysis
  • Figure 3 is a graph showing the frequency difference between groups of major strains in the Firmicutes phylum according to mixed strains and combined administration of herbal medicines according to one embodiment.
  • Figure 4 is a graph showing the frequency difference between groups of major strains in the Bacteroidetes phylum according to mixed strains and combined administration of herbal medicines according to one embodiment.
  • 5 is a graph showing changes in the Firmicutes/Bacteroidetes ratio according to administration of a mixed strain composition according to an embodiment.
  • One aspect includes a first active material including a Lactobacillus sp. strain or a culture thereof and a second active material including herbal medicine as an active ingredient;
  • a first active material including a Lactobacillus sp. strain or a culture thereof and a second active material including herbal medicine as an active ingredient;
  • an anti-aging composition comprising the first active material as an active ingredient and administered in combination with the second active material or a composition for preventing, improving or treating aging-related diseases is provided.
  • the composition further comprises a third active substance including inulin or vitamins; Alternatively, the third active material may be additionally co-administered.
  • the Lactobacillus is an aerobic or facultative anaerobic gram-positive bacillus microorganism widely distributed in nature, and the strain of the genus Lactobacillus is not limited as long as it is a strain of the genus Lactobacillus known in the art. .
  • Lactobacillus genus refers to the old Lactobacillus genus ( Lactobacillus sp . ) means.
  • the strain is Lactobacillus fermentum GB102 belonging to the genus Lactobacillus sp. deposited as KCTC 14105BP, deposited with accession number KCTC 14106BP Lactobacillus sp. It may be Lactobacillus fermentum GB103 strain belonging to or Lactobacillus plantarum GB104 strain belonging to Lactobacillus sp. deposited as KCTC 14107BP.
  • the first active material may further include a Bifidobacterium animalis sub sp. Lactis strain.
  • the term "culture” may be used interchangeably with “culture supernatant”, “conditioned culture medium” or “conditioned medium”, and may supply nutrients so that strains of the genus Lactobacillus can grow and survive in vitro. It may mean the entire medium including the strain obtained by culturing the strain in a medium for a certain period of time, its metabolites, extra nutrients, and the like.
  • the culture solution may mean a culture solution obtained by removing the cells from the cell culture solution obtained by culturing the strain.
  • the medium may be selected from known liquid medium or solid medium, and may be, for example, MRS liquid medium, GAM liquid medium, MRS agar medium, GAM agar medium, and BL agar medium, but is not limited thereto.
  • the second active material may be one or more selected from the group consisting of yam, adlay, red ginseng, ganoderma lucidum, and combinations thereof.
  • the term "herbal medicine” is defined in the Pharmaceutical Affairs Act as raw materials used to prepare herbal medicines or herbal preparations. Herbal medicines may have different medicinal properties depending on the processing method, so the processing method and manufacturing method are important.
  • the herbal medicine may be yam, adlay, red ginseng or ganoderma lucidum, but is not limited thereto. At this time, the herbal medicine may be used in the form of powder or extract.
  • yam Dioscorea japonica
  • Dioscoeaceae a perennial vine plant of the genus Dioscoeaceae , and about 650 species of 10 genera are currently known, but about 10 of these are used as food resources. It grows mainly in mountainous areas and is distributed in Korea and Japan.
  • the rhizome contains amylase, betasin, mucin, mucilage, allantoin, saponin, tannin, polyphenol, etc., and antioxidant, anti-inflammatory, and anticancer effects have been reported.
  • the term "coix lachryma-jobi var. mayuen” is a plant belonging to the Poaceae family and is mainly cultivated in Southeast Asia. When used as a herbal medicine, it is called Uiyiin, and it is used for edema, bladder stones, and rheumatism because it has diuretic, analgesic, and tonic effects. In addition, the raw leaves are used as a substitute for tea, and the root is used to treat jaundice and neuralgia. Recently, it has been known to have anti-cancer effects.
  • red ginseng is steamed roots of ginseng ( Panax ginseng CA Meyer ). Anti-aging, anti-fatigue and anti-stress effects, blood circulation, immune function, prevention of osteoporosis, treatment of anemia, treatment of male infertility, improvement of hypertension and diabetes, prevention of cancer, etc. are known.
  • the red ginseng may be in liquid or powder form.
  • Ganoderma lucidum refers to Ganoderma lucidum (Leyss. ex. Fr) Karst, which is also called Seoncho, Auspicious Mushroom, and Jeokji. It is rich in minerals such as calcium and potassium and contains a lot of unsaturated fatty acids, so it is known to be effective in preventing adult diseases, removing phlegm, and relieving asthma. In addition, the effect of suppressing the growth of cancer, lowering cholesterol content in blood vessels, and lowering blood pressure is also known.
  • the herbal medicine of the present invention may be a mixture of one or more kinds.
  • the herbal medicine may be a mixture of two kinds of herbal medicine.
  • the herbal medicine may be a mixture of yam and adlay.
  • the herbal medicine may be a mixture of yam and red ginseng.
  • the herbal medicine may be a mixture of yam and ganoderma lucidum.
  • the herbal medicine may be a mixture of adlay and red ginseng.
  • the herbal medicine may be a mixture of adlay and ganoderma lucidum.
  • the herbal medicine may be a mixture of red ginseng and ganoderma lucidum.
  • the herbal medicine may be a mixture of three kinds of herbal medicine.
  • the herbal medicine may be a mixture of yam, adlay and red ginseng.
  • the herbal medicine may be a mixture of yam, adlay, and ganoderma lucidum.
  • the herbal medicine may be a mixture of yam, red ginseng and ganoderma lucidum.
  • the herbal medicine may be a mixture of adlay, red ginseng and ganoderma lucidum.
  • the herbal medicine may be a mixture of four kinds of herbal medicine.
  • the four kinds of herbal medicines of the yam, adlay, red ginseng, and ganoderma lucidum may be mixed in a predetermined ratio.
  • the third active material may be one or more selected from the group consisting of inulin, vitamin B2, and combinations thereof.
  • inullin is a polysaccharide contained in many root vegetables and is a type of dietary fiber known as fructan. It is used by some plants as a means of storing energy. After ingestion, inulin is not decomposed by digestive enzymes in the body, but is fermented by intestinal microorganisms to promote bowel function and then discharged out of the body. Effects such as suppression of postprandial rise in blood sugar, improvement of cholesterol and relief of constipation are known.
  • vitamin B2 is referred to as riboflavin or lactoflavin, and is a type of water-soluble vitamin. It is contained in liver, egg yolk, spinach, milk, etc., and deficiency can cause symptoms such as growth retardation, premature aging, pseudomembranitis, dermatitis, alopecia, sore throat, and gastrointestinal disorders.
  • the term "included as an active ingredient” means that a strain of the genus Lactobacillus, an endoplasmic reticulum derived from the strain, a lysate of the strain, a culture medium, or an extract of a culture medium thereof are added, and various drugs are used for drug delivery and stabilization. It means that it is formulated in various forms by adding ingredients as subcomponents.
  • the term "therapeutically effective amount” refers to a method of the present invention or herbal medicine for the method and use of the present invention that elicits a biological or medical response or a desired therapeutic effect in a patient to be obtained by a researcher, doctor, or other clinician. And the amount of the pharmaceutical composition containing herbal medicine for use.
  • a therapeutically effective amount of an herbal medicine may vary depending on factors such as the disease state, age, sex, and weight of the subject, and the ability of the herbal medicine to elicit a desired response in the subject.
  • a therapeutically effective amount is also one in which the therapeutically beneficial effects outweigh any toxic or detrimental effects.
  • the first active material may include a strain of the genus Lactobacillus alone as an active ingredient, or may include one or more pharmaceutically acceptable carriers, excipients, or diluents.
  • the second active material may include herbal medicine alone as an active ingredient, or may include one or more pharmaceutically acceptable carriers, excipients, or diluents.
  • the first active material is Lactobacillus ferementum and Lactobacillus plantarum
  • the second active material is yam, adlay, red ginseng and ganoderma lucidum
  • the active material additionally includes a Bifidobacterium animalis sub sp. Lactis strain and a Lactobacillus acidophilus strain, and additionally contains inulin and vitamins as the third active material it could be
  • Oxidative damage underlies the pathophysiological mechanism that causes sarcopenia (and other senile diseases), and interventions to enhance endogenous antioxidant defenses (eg, administration of antioxidants) result in inhibition of aging.
  • sarcopenia and other senile diseases
  • interventions to enhance endogenous antioxidant defenses eg, administration of antioxidants
  • resveratrol a polyphenolic compound contained in red wine
  • oxidative stress has been shown to increase bone turnover by causing an imbalance between cells responsible for bone formation and resorption.
  • combined administration of mixed strains of the genus Lactobacillus and/or herbal medicines can improve muscle strength decreased due to aging. Specifically, in one embodiment of the present invention, it was confirmed that the grip strength increased in the group administered with the mixed strain and/or herbal medicine mixture to aged mice.
  • anti-aging includes delaying or preventing aging of a cell or object, or converting a senescent cell into a younger cell.
  • the anti-aging may be one or more selected from the group consisting of anti-aging of skeletal muscle cells, anti-aging of nerve cells, anti-aging of skin cells, anti-aging of immune cells, and anti-aging of the intestinal microbial environment. there is.
  • aging-related diseases may be muscle aging-related diseases (eg, sarcopenia) or obesity.
  • the term “combination therapy” or “combined administration” or “in combination” refers to any form of simultaneous or concurrent treatment using at least two separate therapeutic agents.
  • the components of the combination therapy may be administered simultaneously, sequentially or in any order.
  • the ingredients can be administered in a suitable manner, in different doses or at different frequencies of administration or via different routes.
  • the combined administration may be the simultaneous administration of a strain of the genus Lactobacillus and herbal medicine, or the administration of a strain of the genus Lactobacillus followed by administration of herbal medicine.
  • Combination therapy according to the present invention is such that efficacy, measured by, for example, the degree of response, rate of response, time to disease progression or survival time, is the efficacy that can be obtained by administering one or the other of the components of the combination therapy at conventional doses. More therapeutically superior can be defined as capable of providing a synergistic effect.
  • the efficacy of combination therapy is synergistic if the efficacy is therapeutically superior to the efficacy obtained using each of the above alone.
  • administered simultaneously is not particularly limited and means that the components of the combination therapy are administered substantially simultaneously, eg as a mixture or in an immediately succeeding sequence.
  • the term "sequentially administered” is not particularly limited, and means that the components of the combination therapy are not administered simultaneously, but are administered one by one or in groups with a specific time interval between administrations.
  • the time interval may be the same or different between administration of each of the components of the combination therapy, and may be selected, for example, from the range of 2 minutes to 96 hours, 1 day to 7 days or 1 week, 2 weeks or 3 weeks. .
  • the time interval between administrations may range from several minutes to several hours, for example from 2 minutes to 72 hours, from 30 minutes to 24 hours, or from 1 to 12 hours. Additional examples include time intervals ranging from 24 to 96 hours, 12 to 36 hours, 8 to 24 hours, and 6 to 12 hours.
  • the pharmaceutical composition further comprises a third active substance including inulin or vitamins; Alternatively, the third active material may be additionally co-administered.
  • first active substance In the pharmaceutical composition, “first active substance”, “second active substance”, “third active substance”, “anti-aging” and “combined administration” are as described above.
  • prevention may refer to any action that suppresses or delays the onset of a disease state of a subject by administration of a pharmaceutical composition according to one aspect.
  • treatment may refer to any activity in which symptoms of a disease state of an individual are improved or beneficially changed by administration of a pharmaceutical composition according to one aspect.
  • muscle aging refers to the gradual weakening of muscle density and function as mitochondria in muscle fibers lose their activity or the number of mitochondria decreases due to aging, including sarcopenia. it means to
  • muscle aging-related disease refers to diseases caused by changes in the state of muscles and abnormalities caused by aging.
  • muscle loss due to aging is also referred to as age-related sarcopenia. do.
  • the muscle-related disease is sarcopenia, geriatric sarcopenia, atony, muscular atrophy, muscular dystrophy, disuse muscle atrophy, movement motor neuron disease, inflammatory myopathy, neuromuscular junction disease, endocrine myopathy, muscle degeneration, myoclonus, amyotrophic sclerosis, myasthenia gravis, myositis, muscle calcification, muscle ossification, muscle weakness-related diseases and cachexia It may, but is not limited thereto.
  • the above muscle aging-related diseases may be accompanied by muscle inflammation due to trauma, and the composition of the present invention has an effect of alleviating or improving muscle inflammation.
  • the obesity is caused by not only weight gain, but also overeating, excessive drinking and bulimia, hypertension, diabetes, increased plasma insulin concentration, insulin resistance, hyperlipidemia, metabolic syndrome, insulin resistance syndrome, obesity-related gastroesophageal reflux, arteriosclerosis, hypercholesterolemia, uric acid It can lead to obesity-related diseases such as hyperemia, cardiac hypertrophy and left ventricular hypertrophy, lipodystrophy, non-alcoholic steatohepatitis, cardiovascular disease or polycystic ovary syndrome. Therefore, the composition can be used for prevention or treatment of not only obesity but also obesity-related diseases. In addition, the composition can be used for subjects who have a desire to lose weight even if they are not obese.
  • the obesity may be due to various causes.
  • the cause may be a high-fat diet, reduced exercise, genetics, psychological factors, endocrine abnormalities, metabolic abnormalities, social and environmental factors.
  • the obesity may be induced by a high-fat diet.
  • the composition according to one embodiment may include 0.001% by weight to 80% by weight of Lactobacillus strains based on the total weight of the composition.
  • the dosage of the Lactobacillus genus strain may be 0.01 mg to 10,000 mg, 0.1 mg to 1000 mg, 1 mg to 100 mg, 0.01 mg to 1000 mg, 0.01 mg to 100 mg, 0.01 mg to 10 mg, or 0.01 mg to 1 mg.
  • 1X10 3 to 1X10 16 CFU/g of live or dead cells may be divided and administered once or several times.
  • the dosage may be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate and reaction sensitivity, and those skilled in the art can Dosage can be appropriately adjusted in consideration of these factors.
  • the number of administrations can be once or twice or more within the range of clinically acceptable side effects, and administration can be performed at one or two or more sites.
  • the same dosage per kg (body weight) as for humans is administered, or the above-described administration is performed at the volume ratio (eg, average value) of the organ (heart, etc.) between the target animal and the human.
  • the amount converted into the amount can be administered.
  • Possible routes of administration include oral, sublingual, parenteral (eg, subcutaneous, intramuscular, intraarterial, intraperitoneal, intrathecal, or intravenous), rectal, topical (including transdermal), inhalation, and injection, or implantable devices. or the insertion of substances.
  • animals to be treated according to one embodiment include humans and other target mammals, and specifically include humans, monkeys, mice, rats, rabbits, sheep, cows, dogs, horses, and pigs.
  • the composition includes a killed dried strain, and can be administered at 1 g to 10 g, 0.5 g to 1.5 g, 2.5 g to 3.5 g, or 4.5 g to 5.5 g once a day, once to 3 times a day. Can be administered twice.
  • a pharmaceutical composition according to one embodiment may include a pharmaceutically acceptable carrier and/or additives.
  • a pharmaceutically acceptable carrier and/or additives for example, sterile water, physiological saline, common buffers (phosphoric acid, citric acid, other organic acids, etc.), stabilizers, salts, antioxidants (ascorbic acid, etc.), surfactants, suspending agents, tonicity agents, or preservatives, etc. can do.
  • it may also include combining organic substances such as biopolymers, inorganic substances such as hydroxyapatite, specifically collagen matrices, polylactic acid polymers or copolymers, polyethylene glycol polymers or copolymers, and chemical derivatives thereof.
  • Lactobacillus genus bacteria may be dissolved or dispersed in a pharmaceutically acceptable carrier, or frozen in a dissolved or dispersed solution state. .
  • the dosage form may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or in the form of a powder, granule, tablet or capsule.
  • the pharmaceutical composition is administered in a pharmaceutically effective amount.
  • pharmaceutically effective amount means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level depends on the type, severity, drug activity, It may be determined according to factors including sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, drugs used concurrently, and other factors well known in the medical field.
  • the composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or in multiple doses. Considering all of the above factors, it is important to administer the amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.
  • Another aspect includes a first active material including a Lactobacillus sp. strain or a culture thereof and a second active material including herbal medicine as an active ingredient; Or it provides a health functional food for preventing or improving muscle-related diseases or obesity, which contains the first active substance as an active ingredient and is co-administered with the second active substance.
  • Another aspect includes a first active material including a Lactobacillus sp. strain or a culture thereof and a second active material including herbal medicine as an active ingredient; Or it provides a food composition for preventing or improving muscle-related diseases or obesity, which contains the first active substance as an active ingredient and is co-administered with the second active substance.
  • the health functional food further comprises a third active substance including inulin or vitamins; Alternatively, the third active material may be additionally co-administered.
  • first active substance In the health functional food, "first active substance”, “second active substance”, “third active substance”, “anti-aging”, “muscle-related disease”, “metabolic disease”, “obesity” and “combination” Administration "is as described above.
  • the health functional food may further include a food-acceptable carrier.
  • the term "acceptable in food science” means that it exhibits characteristics that are not toxic to cells or humans exposed to the compound.
  • the term “improvement” may refer to any activity that at least reduces a parameter related to the condition being treated, eg, the severity of a symptom.
  • the health functional food may be used before or after the onset of the disease, simultaneously with or separately from a drug for treatment, for the prevention or improvement of cancer.
  • the active ingredient may be added to food as it is or used together with other food or food ingredients, and may be appropriately used according to conventional methods.
  • the mixing amount of the active ingredient can be suitably determined depending on the purpose of its use (for prevention or improvement).
  • the health functional food may be added in an amount of about 15% by weight or less, more specifically about 10% by weight or less, based on the raw material.
  • the amount may be less than the above range.
  • carrier examples include lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium phosphate, calcium silicate, and microcrystalline cellulose.
  • natural flavoring agents thaumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)
  • synthetic flavoring agents sacharin, aspartame, etc.
  • the ratio of the natural carbohydrates may be appropriately determined by a person skilled in the art.
  • the health functional food is various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and salts thereof , alginic acid and its salts, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like.
  • vitamins, minerals electrophilic acids
  • flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.
  • pectic acid and salts thereof alginic acid and its salts
  • organic acids protective colloidal thickeners
  • pH regulators pH regulators
  • stabilizers stabilizers
  • preservatives glycerin
  • alcohol carbonating agents used in carbonated beverages, and the like.
  • the health functional food may be provided in a mixture with conventionally known health functional food for preventing or improving muscle-related diseases or metabolic diseases or other existing health functional foods, and preventing or improving other muscle-related diseases or metabolic diseases.
  • Functional health food for use may be conventionally known health functional food for preventing or improving metabolic diseases, existing health functional food, or newly developed health functional food.
  • the health functional food includes other health functional foods having an effect of preventing or improving muscle-related diseases or metabolic diseases, it is important to mix the amount that can obtain the maximum effect with the minimum amount without side effects, which is known to those skilled in the art. can be easily determined by
  • the food composition for preventing or improving muscle-related diseases and obesity includes all types of functional foods, nutritional supplements, health foods, and food additives,
  • the food composition of can be prepared in various forms according to conventional methods known in the art.
  • compositions herein may be considered food supplements.
  • Food supplements also known as dietary supplements or nutritional supplements, can be considered another particular pharmaceutical product. It is intended to supplement the diet and is intended to provide nutrients or beneficial ingredients that may not be consumed or in sufficient quantities in a normal diet.
  • Most dietary supplements are considered foods, but sometimes they are considered drugs, natural health products or nutraceutical products.
  • food supplements include health functional foods. Food supplements are usually sold over the counter without a prescription. When dietary supplements take the form of pills or capsules, they contain the same excipients used in pharmaceuticals. However, dietary supplements may also be taken in the form of foods fortified with some nutrients (eg infant formula).
  • a composition of the present invention is a food supplement.
  • compositions according to the present invention may be administered as such or mixed with suitable edible liquids or solids or administered as tablets, pills, capsules, lozenges, granules, powders ), suspensions, sachets, syrups, or may be freeze-dried in unit dose form. It may also be in the form of monodoses of the lyophilized composition to be mixed in a separate liquid container provided with it prior to administration.
  • the composition of the present invention may be included in various edible foods and foods, such as milk products in the case of infants.
  • the term "edible product” includes, in its broadest sense, any product in any form capable of being consumed by an animal (e.g., a product received by the sense organs). products that can be imported).
  • the term “food product” is understood to be an edible product that supplies nutritional support to the body. Foods of particular interest are food supplements and infant formulas.
  • the food preferably contains oatmeal gruel, lactic acid fermented foods, resistant starch, dietary fibers, carbohydrates, proteins and glycated proteins. (carrier material) such as (glycosylated proteins).
  • the bacterial cells of the present invention are homogenized with other ingredients such as cereals or powdered milk to constitute an infant formula.
  • Another aspect includes a first active material including a Lactobacillus sp. strain or a culture thereof and a second active material including herbal medicine as an active ingredient; Or, it provides a feed composition for preventing or improving muscle-related diseases or obesity, which includes the first active material as an active ingredient and is co-administered with the second active material.
  • the feed composition may further include one or more strains selected from the group consisting of Bifidobacterium animalis lactis strain and Lactobacillus acidophilus strain, or a culture thereof.
  • the composition may further include inulin and vitamin B2.
  • the Lactobacillus genus strain, culture, herbal medicine, inulin, vitamin B2, aging-related diseases, prevention and treatment are the same as described above.
  • the feed composition for preventing or improving aging-related diseases can be prepared by adding the strain or its culture and herbal medicine in an appropriate effective concentration range according to various feed manufacturing methods known in the art, and preventing or improving aging-related diseases. It can be used as a feed additive composition for this purpose.
  • the “feed” may mean any natural or artificial diet, meal, etc., or component of the meal, intended for or suitable for consumption by an animal.
  • the type of feed is not particularly limited, and feeds commonly used in the art may be used.
  • Non-limiting examples of the feed include vegetable feeds such as grains, root fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal or grain by-products: proteins, fats, and animal feeds such as oils and fats, mineral oils, fats and oils, unicellular proteins, zooplankton, or food.
  • Lactobacillus fermentum Lactobacillus fermentum
  • Lactobacillus plantarum Lactobacillus plantarum
  • the Lactobacillus fermentum strain and the Lactobacillus plantarum strain of the present invention were isolated from vaginal samples of healthy women who visited a hospital for the purpose of health examination. Specifically, vaginal internal samples were taken with a cotton swab, pre-inoculated on a Rogosa SL (MRS) plate medium, and cultured for 48 hours in an anaerobic chamber at 37°C. When bacterial colonies grew, single colonies were subcultured onto fresh MRS plates for pure isolation. After pure separation, strain culture was performed using MRS medium.
  • MRS Rogosa SL
  • pancreatic lipolysis enzyme lipase
  • pancreatic lipolytic enzyme Specifically, the ability to inhibit the activity of pancreatic lipolytic enzyme was obtained by diluting the strain of 1-1 to a concentration of 0.1 mg/mL, followed by 0.167 mM p-nitrophenylpalmitate (PNP; Sigma, USA) solution, 0.061 M Tris-HCl buffer (pH 8.5), put it on a plate with 0.3 mg/mL lipase solution, reacted at 25°C for 10 minutes, and then measured absorbance at 405 nm.
  • PNP p-nitrophenylpalmitate
  • 3T3-L1 cell viability of the strain obtained in 1-1 was measured using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method.
  • 3T3-L1 cells were dispensed into a 96-well plate at a concentration of 16 ⁇ 10 4 cells/well, and the culture medium was removed after 24 hours of culture.
  • lactic acid bacteria samples diluted by concentration 100, 1,000 ⁇ g/mL
  • 20 ⁇ L of a 5 mg/mL MTT (Sigma, USA) solution was added, and incubated at 37° C. for 4 hours.
  • the supernatant was removed, 200 ⁇ L of dimethyl sulfoxide (DMSO) was added, and absorbance was measured at 546 nm.
  • DMSO dimethyl sulfoxide
  • Lactobacillus fermentum GB102 (hereinafter referred to as 'GB102') and Lactobacillus fermentum GB103 showed low cytotoxicity and inhibitory effect on fat cell accumulation.
  • 'GB103' Lactobacillus plantarum GB104
  • 'GB104' Lactobacillus plantarum GB104
  • Lactobacillus fermentum was changed to Limosilactobacillus fermentum
  • the strain name was changed to Lactobacillus plantarum
  • Lactobacillus plantarum was changed to Lactiplantibacillus plantarum . name has been changed
  • the modified strain names of the existing strains were interchangeably described.
  • 16S rRNA gene sequences were analyzed.
  • the 16S rRNA gene sequence obtained through PCR using 27F and 1492R primers targeting the bacterial 16S rRNA gene was analyzed by the Sanger sequencing method, and Lactobacillus fermentum GB102, GB103 and Lactobacillus plantarum were analyzed.
  • the 16s rRNA sequences of GB104 are shown as Sequence Listings 1, 2 and 3, respectively.
  • GB104 is a strain belonging to Lactiplantibacillus plantarum through comparison of genome sequence average similarity (Average Nucleotide Identity, ANI) values after genome decoding.
  • ANI Average Nucleotide Identity
  • Lactobacillus fermentum Lactobacillus fermentum
  • next-generation sequencing technology NGS
  • bioinformatics technology were used to completely decipher the genome sequence of the strain and include it in the genome.
  • the functions of the genes were inferred.
  • specificity of the strain was confirmed through comparative analysis with the completely translated genome sequence of the same species.
  • This strain was cultured for 4 hours in anaerobic conditions at 37 ° C in MRS liquid medium, and genomic DNA was extracted from the culture using the MG Genomic DNA purification kit (MGMED, Inc., Korea).
  • Sequence analysis was performed using PacBio RS II equipment to obtain long read data with an average length of 10 kb or more, and sequence length of less than 500 bp with high accuracy to complement long read data with low accuracy Sequence analysis was performed using NovaSeq 6000 equipment to produce short read data.
  • the long lead data was assembled into a highly complete GB102 and GB103 genome draft using the SMRT Analysis server's HGAP2 pipeline. SNP and InDel errors that may exist in the assembled draft genome sequence were corrected through long read data and short sequence data.
  • CDS coding sequences
  • rRNA and tRNA were predicted using the RFAM tool.
  • Predicted CDSs were predicted by performing a homology search (applying the BLAST algorithm) based on the open UniProt database, GenBank nr database, Subsystem database, PFAM database, and COG database.
  • the two strains GB102 and GB103 showed an ANI value of 95% or more compared to the Lactobacillus fermentum standard strain, proving that they belong to the Lactobacillus fermentum species (Table 2).
  • the GB102 and GB103 strains are new species that have not been previously isolated and reported because they do not match 100% with the standard strain.
  • strains GB102 and GB103 were also identified as different strains due to the large genomic distance.
  • the present inventors named the GB102 and GB103 strains as “ Lactobacillus fermentum GB102” (Accession No.: KCTC 14105BP) and “ Lactobacillus fermentum GB103” (Accession No.: KCTC 14106BP) and named them as Korea Each was deposited at the Korean collection for type cultures (KCTC) at the Research Institute of Bioscience and Biotechnology on January 14, 2020.
  • sequence analysis was performed using PacBio RS II equipment, and fragment sequence data with a sequence length of less than 500 bp with high accuracy was produced to supplement long read data with low accuracy.
  • sequence analysis was performed using a NovaSeq 6000 instrument.
  • the long lead data was assembled into a highly complete GB104 genome draft using the SMRT Analysis server's HGAP2 pipeline. SNP and InDel errors that may exist in the assembled draft genome sequence were corrected through long read data and short sequence data.
  • CDSs were predicted from the completed genome sequence using the Prodigal program, and rRNA and tRNA were predicted using the RFAM tool.
  • Predicted CDSs were predicted by performing a homology search (applying the BLAST algorithm) based on the open UniProt database, GenBank nr database, Subsystem database, PFAM database, and COG database.
  • the genome of GB104 is summarized in Table 3.
  • this strain possessed the plantaricin gene group, which is a group of bacteriocin biosynthetic genes related to antibacterial activity found in Lactobacillus plantarum .
  • plantaricin gene group which is a group of bacteriocin biosynthetic genes related to antibacterial activity found in Lactobacillus plantarum .
  • it can be seen that it is a new species that has not been previously isolated and reported because it does not match 100% with the standard strain and reference strain.
  • GB104 bacteria as " Lactobacillus plantarum GB104" (Accession Number: KCTC 14107BP) and submitted it to the Korean cell line bank (Korean collection for type cultures, KCTC), Korea Research Institute of Bioscience and Biotechnology, on January 2020. Deposited on the 14th of the month.
  • DPPH free radical scavenging activity was evaluated in order to confirm the antioxidant ability of materials containing GB102, GB103 and GB104 strains as active ingredients or strains and auxiliary ingredients (vitamin B2 and red ginseng).
  • the DPPH solution was prepared to be a 0.2 mM solution in 100% methanol.
  • strains GB102, GB103, and GB104 and Lacticaseibacillus rhamnosus GG (formerly Lactobacillus rhamnosus GG , positive control) strains were inoculated into MRS broth and incubated at 37° C. for 16 hours. Thereafter, the strain culture medium was centrifuged (3600 rpm, 15 minutes) to obtain cultured lactic acid bacteria, washed twice with 1X PBS, and a mixture of three lactic acid bacteria of GB102, GB103, and GB104 5x10 9 CFU / mL concentration samples were prepared.
  • a comparative strain of Lacticaseibacillus rhamnosus GG ( Lacticaseibacillus rhamnosus GG) was also prepared at a concentration of 5x10 9 CFU/ml.
  • samples were prepared with vitamin B2 and red ginseng capsule contents at a concentration of 625 ug/mL, and L-ascorbic acid (positive control) was prepared at a concentration of 12.5 ug/mL.
  • DPPH radical scavenging activity was calculated by the following method.
  • OD control Absorbance of PBS
  • OD sample Absorbance of sample
  • Lacticaseibacillus rhamnosus GG Lacticaseibacillus rhamnosus GG 1x10 9 CFU is about 26.2%
  • L-ascorbic acid L-ascorbic acid
  • 125 ug additives showed about 14%
  • GB102, GB103, GB104 three lactic acid bacteria mixed 1x10 9 CFU was about 44.4%
  • GB102, GB103, GB104 three lactic acid bacteria mixed 1x10 9 CFU + additives 125 ug had about 56.2% efficacy.
  • the mixture of three lactic acid bacteria, GB102, GB103, and GB104 had superior antioxidant efficacy compared to the positive control lactic acid bacteria, Lacticaseibacillus rhamnosus GG.
  • the natural process of aging makes the body more susceptible to oxidative damage.
  • the anti-aging effect is mostly related to its antioxidant properties and free radical scavenging ability.
  • oxidative damage has been proposed as one of the major causes of skeletal muscle decline that occurs with aging.
  • the identification of free radicals as accelerators of the aging process may mean that their inhibition may limit the detrimental modifications they exert on organisms (particularly skeletal muscle). In other words, if molecules with antioxidant capacity are able to counteract oxidative damage, this could play an important role in preventing the development of aging-related conditions, including the neutralization process.
  • Oxidative damage underlies the pathophysiological mechanism that causes sarcopenia (and other senile diseases), and interventions to enhance endogenous antioxidant defenses (eg, administration of antioxidants) result in inhibition of aging.
  • sarcopenia and other senile diseases
  • interventions to enhance endogenous antioxidant defenses eg, administration of antioxidants
  • resveratrol a polyphenolic compound contained in red wine
  • oxidative stress has been shown to increase bone turnover by causing an imbalance between cells responsible for bone formation and resorption.
  • DPPH free radical scavenging activity was evaluated to confirm the antioxidant capacity of each of GB102, GB103 and GB104 strains.
  • a DPPH 0.2 mM solution was prepared using DPPH (Alfa Aesar) reagent in 100% methanol.
  • DPPH Alfa Aesar
  • GB102, GB103, GB104 strains and Lacticaseibacillus rhamnosus GG (positive control) strains were inoculated into MRS broth and incubated at 37°C for 16 hours. Thereafter, the strain culture was centrifuged (3600 rpm, 15 minutes) to obtain cultured lactic acid bacteria, washed twice with 1X PBS, and a sample of GB102, GB103, and GB104 mixed 5x10 9 CFU / ml concentration of three lactic acid bacteria was prepared.
  • Lacticaseibacillus rhamnosus GG Lacticaseibacillus rhamnosus GG Comparative strains were also prepared at a concentration of 5x10 9 CFU/ml. 1x10 9 CFU (200 ul) of GB102, GB103, and GB104 lactic acid bacteria, respectively, and 1x10 9 CFU (200 ul) of a mixture of three lactic acid bacteria were mixed with 200 ul of a 0.2 mM DPPH solution and reacted in a dark room at room temperature for 30 minutes.
  • the auxiliary ingredients co-administered with the mixed strains included herbal medicines, inulin, and vitamin B2, and Andong yam, adlay, red ginseng powder, and ganoderma lucidum were used as herbal medicines.
  • the total content of herbal medicines (Andong yam, adlay, red ginseng powder, ganoderma lucidum), inulin, and vitamin B2 was 70.20 mg, and supplementary materials were administered at a total dose of 100.5 mg per mouse, including 30.30 mg of cryopreservative.
  • Experimental group 1 is a mixed strain consisting of Lactobacillus fermentum GB102 and GB103 strains (Accession No. KTCT 14105BP, KCTC 14106BP) and Lactobacillus plantarum GB104 strain (Accession No. KCTC 14107BP).
  • #7 lactic acid bacteria was orally administered.
  • the mixed strain #7 lactic acid bacteria was administered at a dose of 5X10 8 CFU per mouse, and to maintain the guaranteed number of bacteria, Bifidobacterium animalis lactis ( Bifidobacterium animalis sub sp.
  • Lactis strain and Lactobacillus Acidophilus ( Lactobacillus acidophilus ) was administered together.
  • Experimental group 2 (G2) was orally administered a herbal medicine mixture consisting of Andong yam, adlay, red ginseng powder, and ganoderma lucidum, and auxiliary ingredients including inulin and vitamin B2. Sub-materials were administered at a dose of 100.5 mg per mouse as shown in Table 6 above.
  • experimental group 3 (G3), mixed strains and supplementary materials were administered orally.
  • Mice fed a normal chow diet (NCD) were set as a control. Each control group and experimental group used 15 mice.
  • the experimental conditions of the above experimental group and control group are summarized in Table 7 below.
  • grip strength was measured by holding the wire part of the Grip Strength device with the front two legs of the mouse using the Grip Strength device to confirm the improvement in muscle strength, and the results are shown in the table. 8.
  • DGF-8/myostatin DuoSet (R&D systems) ELISA was used, and detailed experimental methods were performed based on the protocol provided by the manufacturer. Briefly, the analysis method was commonly described, the capture antibody of the product was incubated in a 96-well flat immunoplate (SPL) overnight at 4 ° C the day before, and then washed three times using 0.05% Tween 20 in PBS wash buffer the next day. Thereafter, blocking is performed at room temperature for 1 hour using 1% BSA in PBS sample diluent and washing is performed three times using wash buffer.
  • SPL 96-well flat immunoplate
  • Group PBS mixed strains supplementary material Mixed Strains + supplementary material Mean myostatin (ng/mL) 4689.8 ⁇ 364.7 4439.7 ⁇ 251.2 4363.2 ⁇ 200.6 3721.8 ⁇ 173.2
  • mice A total of three groups were formed to analyze changes in the intestinal flora, anti-aging and anti-obesity activities induced by the combined administration of lactic acid bacteria mixed strains and/or herbal medicines in aged mice.
  • a group (15 mice) orally administered with phosphate-buffered saline (PBS) to 16-month-old C57BL/6 aging mice for 32 weeks and a group of 10-week-old C57BL/6 young mice (10 mice) were set as the control group.
  • PBS phosphate-buffered saline
  • mice 10-week-old C57BL/6 young mice
  • the amplified product was purified using AMPure XP beads (Beckman). Then, large-capacity sequencing data was produced from the purified products using the MiSeq (Illumina) platform. Denoising and paired sequence assembly of large-capacity sequences produced using the DADA2 program were performed. Sequence assembly using the published Mothur pipeline (Schloss PD et al. 2009. Introducing mothur: Open-source, platform-independent, community-supported software for describing and comparing microbial communities. Applied and Environmental Microbiology 75:7537-7541) OTUs clustering and biodiversity analysis of the sequence data were performed. The results are shown in Figures 1 to 5.
  • the Firmicutes/Bacteroidetes ratio which is known as an obesity-related indicator of intestinal flora
  • the combined administration of the mixed strain of lactic acid bacteria and herbal medicines according to one embodiment changes the intestinal flora of the aging mouse to be similar to that of the young mouse, thereby preventing or treating aging-related diseases such as obesity caused by the intestinal flora of the aging mouse. It means what you can give.
  • the mixed strain of lactic acid bacteria and herbal medicines were administered to aged mice, it was found that the ratio tended to be lower than that of the aging mouse control group.

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Abstract

La présente invention concerne une composition anti-âge adaptée pour administrer en association une souche de Lactobacillus sp. et une substance à base de plantes médicinales. Selon un aspect de l'invention, une composition anti-âge conçue pour administrer une souche de Lactobacillus sp. et une substance à base de plantes médicinales associées peut être avantageusement utilisée pour prévenir ou traiter des maladies liées au vieillissement en atténuant les symptômes de faiblesse musculaire provoqués par le vieillissement ou en réduisant le changement induit par le vieillissement dans l'environnement microbien intestinal.
PCT/KR2022/009477 2021-06-30 2022-06-30 Composition anti-âge contenant une souche de lactobacillus sp. et une substance à base de plantes médicinales pour thérapie combinée Ceased WO2023277636A1 (fr)

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