WO2021194011A1 - Probiotiques pour prévenir ou traiter la maladie d'alzheimer, et composition alimentaire les comprenant pour prévenir ou traiter une maladie buccale - Google Patents

Probiotiques pour prévenir ou traiter la maladie d'alzheimer, et composition alimentaire les comprenant pour prévenir ou traiter une maladie buccale Download PDF

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WO2021194011A1
WO2021194011A1 PCT/KR2020/007649 KR2020007649W WO2021194011A1 WO 2021194011 A1 WO2021194011 A1 WO 2021194011A1 KR 2020007649 W KR2020007649 W KR 2020007649W WO 2021194011 A1 WO2021194011 A1 WO 2021194011A1
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strain
oral
composition
smfm2016
alveolar bone
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Korean (ko)
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윤요한
최경희
최유경
박은영
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숙명여자대학교산학협력단
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • 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
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/312—Foods, ingredients or supplements having a functional effect on health having an effect on dental health
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus

Definitions

  • Lactobacillus curvatus Lactobacillus curvatus
  • SMFM2016-NK strain accession number: KCTC14138BP
  • oral disease prevention or improvement food composition comprising the strain, oral disease prevention or treatment pharmaceutical composition, probiotics, and feed to the composition.
  • Oral disease refers to all diseases that can occur in the oral cavity, including teeth.
  • Common oral diseases include dental caries, that is, periodontal diseases such as tooth decay, periodontitis, and gingivitis. This can be.
  • Periodontal disease includes inflammatory diseases that damage soft tissues such as periodontal ligaments and gingiva surrounding teeth and hard tissues such as alveolar bone. plays a role In alveolar bone, bone formation by osteoblasts and bone resorption by osteoclasts are metabolized. When bone resorption exceeds bone formation due to various causes, the alveolar bone lowers, exposing the tooth roots, and eventually supporting the teeth. cannot be performed. This loss of alveolar bone can also be caused by gingivitis and periodontitis.
  • Gingivitis is an inflammation limited to the soft tissues of the gums and is a disease with relatively light and recoverable symptoms. It causes inflammation and bone loss of the periodontal ligament.
  • Such oral disease or periodontal disease is not only a disease that can occur most often in the adult population, regardless of domestic or foreign, but also a disease of public health interest that requires considerable cost in terms of personal medical care and social management system.
  • the number of periodontal disease patients in Korea between 2012 and 2016 increased from 7.07 million in 2012 to 11.07 million in 2016, an increase of about 56.6% in four years, which corresponds to an average annual growth rate of 12%. Therefore, various surgical treatment methods exist and have been developed for the treatment of such periodontal disease, but for individuals, surgical treatment is not only psychologically reluctant but also carries a huge cost burden, There is a problem that continuous treatment at an appropriate time is difficult because there are cases where it is performed. Therefore, for oral diseases including periodontal disease, it can be said that not only surgical treatment, but also establishment of personal oral hygiene and prevention through this is more important.
  • Lactobacillus curvatus SMFM2016-NK strain decreases the number of osteoclasts, increases the number of osteoblasts, decreases the volume of cancellous bone loss, increases the volume of alveolar bone, reduces the number of inflammatory cells, and harmful effects in the oral cavity
  • the present invention was completed by confirming that it exhibits excellent antibacterial activity against bacteria.
  • probiotics reached about $33 billion (about KRW 36 trillion) in 2015, and is expected to increase by 7% every year to reach about $ 47 billion (about KRW 53 trillion) by 2020.
  • probiotics are highly likely to be used not only as health functional foods using them, but also as quasi-drugs and medicines for oral care and oral disease management, the significance of the present invention is also very large.
  • the present invention aims to solve the above problems and other problems related thereto.
  • An exemplary object of the present invention is to provide a newly isolated and identified Lactobacillus curvatus SMFM2016-NK strain (Accession No.: KCTC14138BP) from kimchi.
  • Another exemplary object of the present invention is to provide a food composition for preventing or improving oral diseases, including the Lactobacillus curbatus SMFM2016-NK strain, its culture, concentrate, dry matter or lysate.
  • Another exemplary object of the present invention is to provide a pharmaceutical composition for preventing or treating oral diseases, including the strain, its culture, concentrate, dry matter or lysate.
  • Another exemplary object of the present invention is to provide a probiotic comprising the above strain, a culture, concentrate, dried product or lysate thereof.
  • Another exemplary object of the present invention is to provide a quasi-drug composition comprising the strain, its culture, concentrate, dried product or lysate.
  • Lactobacillus curvatus (Lactobacillus curvatus) SMFM2016-NK strain.
  • the strain was isolated from kimchi and was deposited at the Korea Institute of Biotechnology and Biotechnology Biological Resources Center (KCTC) on February 17, 2020, and was given an accession number KCTC 14138BP.
  • the strain includes the genomic characteristics of Table 7, FIGS. 5 and 6 and the 16s rDNA sequence shown in SEQ ID NO: 1, newly isolated and identified by the present inventors who confirmed that there is no identical strain as a result of genomic analysis is a strain This corresponds to a probiotic strain, is harmless to the human body, and can be used without side effects.
  • the strain can exhibit the effect of reducing the number of osteoclasts, increasing the number of osteoblasts, decreasing the volume of cancellous bone loss, increasing the volume of alveolar bone, and reducing the number of inflammatory cells in oral teeth or periodontal tissue, etc.
  • Intermedia Prevotella intermedia
  • Fusobacterium nucleatum Fusobacterium nucleatum
  • Aggregatibacter actinomycetemcomitans Aggregatibacter actinomycetemcomitans
  • It may be one that exhibits antibacterial activity against harmful bacteria in the oral cavity.
  • Lactobacillus curbatus SMFM2016-NK strain (Accession No.: KCTC14138BP), its culture, concentrate, dry matter or lysate, including oral disease prevention or improvement food A composition is provided.
  • Lactobacillus ( Lactobacillus ) refers to a microorganism of the genus Gram-positive bacillus of aerobic or facultative anaerobic nature widely distributed in nature, and the description of the Lactobacillus curvatus SMFM2016-NK strain is as described above.
  • the term "culture” refers to a medium containing by-products and strains generated through the ingestion of nutrients and metabolism by culturing the strain in a medium, and the culture of the Lactobacillus curvatus SMFM2016-NK strain. It could mean water.
  • the food composition of the present invention may include a culture of the above strain as well as a concentrate, a dried product, or a lysate thereof.
  • oral disease refers to all diseases that can occur in the oral cavity, including teeth, specifically, a decrease in the number of osteoclasts in the oral tissue, an increase in the number of osteoblasts, a decrease in the volume of cancellous bone loss, and alveolar bone. Includes all diseases that can be prevented, ameliorated and/or treated due to an increase in volume, a decrease in the number of inflammatory cells, and an antibacterial effect on harmful oral bacteria.
  • the oral disease is alveolar bone breakage, alveolar bone osteoporosis, alveolar bone osteomalacia, alveolar bone osteopenia, dental caries, gingivitis ) and periodontitis (periodontitis) may be one or more diseases selected from the group consisting of, but is not particularly limited thereto.
  • the food composition of the present invention includes the Lactobacillus curbatus SMFM2016-NK strain, its culture, concentrate, dry matter or lysate, the number of osteoclasts in the oral cavity or periodontal tissue, etc. decreases, the number of osteoblasts increases, cancellous bone Loss volume reduction, alveolar bone volume increase, and inflammatory cell count reduction effects can be equally represented, and Prevotella intermedia , Fusobacterium nucleatum ) or Aggregatibacter actinomycetemcomy Antibacterial activity against harmful bacteria in the oral cavity, such as Aggregatibacter actinomycetemcomitans, may also be exhibited in the same manner.
  • the strain decreases the number of osteoclasts, increases the number of osteoblasts, decreases the volume of cancellous bone loss, increases the volume of alveolar bone and reduces the number of inflammatory cells, and the specific use thereof as long as it can exhibit antibacterial activity against harmful bacteria in the oral cavity , may be included in any amount (effective amount) depending on the formulation and purpose of mixing, and a typical effective amount will be determined within the range of 0.001 wt % to 50.0 wt % based on the total weight of the composition.
  • the "effective amount” refers to the amount of the active ingredient contained in the composition of the present invention that can exhibit the intended functional and pharmacological effects according to the composition of the present invention to a mammal, preferably a human, to be applied during the administration period by a medical professional or the like. say Such effective amounts can be determined empirically within the ordinary ability of one of ordinary skill in the art.
  • Subjects to which the composition of the present invention can be applied are mammals and humans, particularly preferably humans.
  • the food may include a health functional (sexual) food.
  • the term "health functional food” refers to a food manufactured and processed using raw materials or ingredients having useful functions in the human body.
  • the “functionality” refers to obtaining useful effects for health purposes such as regulating nutrients or physiological actions with respect to the structure and function of the human body.
  • the health functional food of the present invention can be prepared by a method commonly used in the art, and at the time of manufacture, it can be prepared by adding raw materials and components commonly added in the art.
  • the formulation of the health functional food is also recognized as a health functional food, it can be prepared without limitation.
  • the food composition of the present invention can be prepared in various forms, and unlike general drugs, there is no side effect that may occur when taking the drug for a long time using food as a raw material.
  • the food composition of the present invention can be prepared in any form, and specifically, health functional food preparations such as tablets, capsules, pills, granules, liquids, powders, pieces, pastes, syrups, gels, jellies, bars, etc.; It may be one or more formulations selected from the group consisting of beverages, gums and candies, but is not particularly limited thereto.
  • the food composition of the present invention may contain food additives in addition to the active ingredient.
  • Food additives can be generally understood as substances that are mixed or infiltrated with food in manufacturing, processing, or preserving food, and their safety must be ensured as they are consumed daily and for a long period of time with food.
  • the food additives are divided into sweeteners, flavoring agents, preservatives, emulsifiers, acidulants, thickeners, etc. in terms of function, and are not particularly limited as long as the food composition of the present invention meets the intended purpose.
  • the food composition of the present invention may contain physiologically active substances or minerals known in the art for the purpose of functional and nutritional supplementation and guaranteed stability as a food additive in addition to the food additives.
  • the physiologically active substances or minerals are not particularly limited as long as the food composition of the present invention meets the intended purpose.
  • the food additives as described above may be included in an effective amount to achieve the purpose of the addition according to the type of product, and with respect to other food additives that may be included in the food composition of the present invention, the food additives of each country Or you can refer to the Food Additives Ordinance.
  • the term “improvement” refers to the effect of alleviating the symptoms of oral disease or periodontal disease by applying the composition comprising the Lactobacillus curbatus SMFM2016-NK strain of the present invention
  • prevention means any action that suppresses or delays oral disease or periodontal disease-related symptoms by administration of the composition of the present invention.
  • Lactobacillus curvatus Lactobacillus curvatus
  • SMFM2016-NK strain Accession No.: KCTC14138BP
  • Lactobacillus, Lactobacillus curvatus SMFM2016-NK strain, culture and oral disease are the same as described above.
  • the pharmaceutical composition of the present invention includes the Lactobacillus curbatus SMFM2016-NK strain, its culture, concentrate, dry matter or lysate, the number of osteoclasts in the oral cavity or periodontal tissue, etc. decreases, the number of osteoblasts increases, bone loss volume reduction, and equally it represents an increase in bone volume and inflammatory cell count reduction, freebo telra intermediate (Prevotella intermedia), Pew jobak Te Leeum New Clegg Atum (Fusobacterium nucleatum) or Agde Leganes tee bakteo solution Tino My setem Antibacterial activity against harmful bacteria in the oral cavity, such as Aggregatibacter actinomycetemcomitans, may also be exhibited in the same manner.
  • the pharmaceutical composition of the present invention is a pharmaceutically acceptable carrier, excipient or diluent commonly used in the preparation of pharmaceutical compositions in addition to the Lactobacillus curbatus SMFM2016-NK strain, its culture, concentrate, dried product or lysate.
  • the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil may be used, but is not particularly limited thereto.
  • the pharmaceutical composition of the present invention is selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, internal solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solvents and freeze-drying agents according to a conventional method, respectively. It may have any one dosage form, and the dosage form may be in various forms, either oral or parenteral. In the case of formulation, it may be prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants, but is not limited thereto. Specifically, tablets, pills, powders, granules, capsules, etc.
  • the solid preparation may be used in the solid preparation for oral administration, and the solid preparation includes at least one or more excipients such as starch, calcium carbonate, sucrose or lactose, Gelatin or the like may be used.
  • excipients such as starch, calcium carbonate, sucrose or lactose, Gelatin or the like
  • lubricants such as magnesium stearate and talc may be used in addition to simple excipients, but the present invention is not limited thereto.
  • liquid formulations for oral administration suspensions, internal solutions, emulsions, syrups, etc. may be used, and various excipients such as wetting agents, sweetening agents, fragrances, preservatives, etc.
  • a sterile aqueous solution, a non-aqueous solution, a suspension, an emulsion, a freeze-dried formulation, or a suppository may be used as a formulation for parenteral administration.
  • the non-aqueous solvent and suspending agent may include, but are not limited to, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.
  • the optimal amount and dosing interval of individual administration of the pharmaceutical composition of the present invention will be determined by the nature and extent of oral disease or periodontal disease, dosage form, route and site, and the age and health condition of the subject, and the doctor will ultimately use it. It will be appreciated by those skilled in the art that appropriate dosing will be determined. Such dosing may be repeated as often as appropriate, and the dosage and frequency may be altered or reduced according to normal clinical practice.
  • the administration route of the composition of the present invention may be administered through any general route as long as the object of the present invention can be achieved.
  • the pharmaceutical composition of the present invention may be administered intraperitoneally, intravenously, subcutaneously, intradermally, orally, but is not limited thereto.
  • treatment refers to any action in which the symptoms of oral disease or periodontal disease are improved or beneficially changed by administration of the composition of the present invention.
  • Lactobacillus curvatus SMFM2016-NK strain (Accession No.: KCTC14138BP), its culture, concentrate, dry matter or lysate, including probiotics.
  • the term Lactobacillus, Lactobacillus curvatus SMFM2016-NK strain and culture are as described above.
  • the term "probiotics” refers to living microorganisms that provide beneficial effects in the body.
  • the microflora in the oral cavity is improved through the interaction of microorganisms and furthermore, oral disease or periodontal disease is prevented, improved and treated Microorganisms that can achieve the purpose of
  • the strain used as the probiotic for the purpose of the present invention is the Lactobacillus curbatus SMFM2016-NK strain, and may further include other probiotic strains as long as they are effective in preventing, improving and treating oral diseases or periodontal diseases.
  • the probiotics containing the Lactobacillus curbatus SMFM2016-NK strain of the present invention are characterized in that they are used for preventing and improving oral diseases, and the strain reduces the number of osteoclasts in teeth or periodontal tissues in the oral cavity, and the number of osteoblasts. increase, may indicate bone loss volume reduction, increased bone volume and inflammatory cell count reduction, freebo telra intermediate (Prevotella intermedia), Pew jobak Te Leeum New Clegg Atum (Fusobacterium nucleatum) or Agde Leganes tee bakteo solution Tino My setem It may exhibit antibacterial activity against harmful bacteria in the oral cavity, such as Aggregatibacter actinomycetemcomitans.
  • the probiotics of the present invention may be used in the form of Lactobacillus curbatus SMFM2016-NK strain, its culture, concentrate, dried product or crushed product, including powder, fermented milk, granules, capsules, pills, gums, films, etc. It is not particularly limited.
  • Another aspect of the present invention for achieving the above object provides a quasi-drug composition
  • a quasi-drug composition comprising the Lactobacillus curvatus SMFM2016-NK strain (Accession No.: KCTC14138BP), its culture, concentrate, dry matter or lysate.
  • Lactobacillus, Lactobacillus curvatus SMFM2016-NK strain, culture and oral disease are the same as described above.
  • the quasi-drug composition of the present invention includes the Lactobacillus curbatus SMFM2016-NK strain, its culture, concentrate, dry matter, or lysate, the number of osteoclasts in the oral cavity or periodontal tissue, etc. decreases, the number of osteoblasts increases, cancellous bone Loss volume reduction, alveolar bone volume increase, and inflammatory cell count reduction effects can be equally represented, and Prevotella intermedia , Fusobacterium nucleatum ) or Aggregatibacter actinomycetemcomy Antibacterial activity against harmful bacteria in the oral cavity, such as Aggregatibacter actinomycetemcomitans, may also be exhibited in the same manner.
  • the quasi-drug composition of the present invention may include a quasi-drug for oral use.
  • the quasi-drug composition of the present invention may include ingredients commonly used in the composition for oral quasi-drugs, such as abrasives, wetting agents, binders, It may include a foaming agent, a sweetener, a preservative, an active ingredient, a flavoring agent, a colorant, a solvent, a brightener, a solubilizer, or a pH adjuster.
  • the quasi-drug composition of the present invention may be prepared in any formulation conventionally prepared in the art, and specifically, toothpaste, mouthwash, gum, candy, oral spray, oral ointment, oral varnish, It may be one or more formulations selected from the group consisting of oral film and gum massage cream, but is not particularly limited thereto.
  • the quasi-drug composition of the present invention when it is a toothpaste formulation, it may include a wetting agent, abrasive agent, binder, foaming agent, flavoring agent, sweetener, coloring agent, preservative, active ingredient, solvent, pH adjusting agent, and the like.
  • the Lactobacillus curvatus SMFM2016-NK strain (Accession No.: KCTC14138BP) of the present invention is a probiotic strain newly isolated and identified from kimchi, and decreases the number of osteoclasts, increases the number of osteoblasts, decreases the volume of cancellous bone loss, and the volume of alveolar bone. It can be usefully used as a composition for prevention, improvement, and treatment of oral diseases because it has an effect of increasing, reducing the number of inflammatory cells, and exhibiting excellent antibacterial activity against harmful bacteria in the oral cavity.
  • H&E histopathological hematozylin-eosin
  • Figure 4 shows the cell morphological SEM image of the Lactobacillus curbatus SMFM2016-NK strain of the present invention.
  • FIG. 6 shows the analysis results of the unweighted pair group method with arithmetic mean (UPGMA) tree based on the average sequence similarity (ANI %) of the Lactobacillus curbatus SMFM2016-NK strain of the present invention and other Lactobacillus sp. strains of the present invention.
  • UGMA unweighted pair group method with arithmetic mean
  • ANI average sequence similarity
  • Example 1 Lactobacillus curvatus Isolation and identification of SMFM2016-NK strain
  • kimchi collected in Seoul was placed in a sterile filter bag, 100 mL of 0.1% buffered peptone water (BPW) was added, and then homogenized for 1 minute using a pummeler (Bag mixer 400; Interscience Co., Paris, France). 100 ⁇ L of the homogenized solution was taken and spread on MRS agar (Becton, Dickinson and Company, Sparks, MD, USA), and incubated at 35° C. for 24 hours. The isolated strain was inoculated in MRS broth (BD, USA) medium, then cultured at 35°C for 24 hours, and a glycerol stock containing 20% glycerol was prepared and stored at -80°C before use.
  • BPW buffered peptone water
  • the isolated strain was identified through 16s rRNA sequencing. After pure separation of the isolates, the identification was requested to Bionics (Seoul) on MRS agar plate. 16s rRNA sequencing was performed using universal primers 518F (5'-CCAGCAGCCGCGGTAATACG-3') and 800R (5'-TACCAGGGTATCTAATCC-3'). The sequence information was as shown in the attached SEQ ID NO: 1.
  • Example 3 Oral disease animal model
  • Antibiotics (1 mg/mL sulfamethoxazole and 200 ⁇ g/mL trimethoprim in water) were put into a drinking water bottle and ingested for 3 days from the day following the end of the acclimatization period of the experimental animals (Sprague-Dawley rat, male, 6-week-old).
  • Zoletil (1 mg/mL) and Lumpun were mixed in a 4:1 (v/v) ratio and administered intraperitoneally to experimental animals using a syringe at a dose of 1 mL/kg.
  • the lower right first molar was ligated to the boundary between the gum and periodontal area using sterile nylon. After ligation, sterile nylon was pushed to be buried in the gums, and oral administration was performed while maintaining the ligation for 28 days (FIG. 1).
  • lactic acid bacteria glycerol stock solution 100 ⁇ L was inoculated into a 50 mL tube containing 45 mL of MRS broth (Becton, Dickinson and Company), and incubated at 35 °C for 18-24 hours.
  • SD periodontitis-induced Sprague-Dawley
  • Example 4 Micro-CT analysis and cancellous bone loss volume analysis
  • micro-CT was measured by setting a predetermined periodontitis-induced site, and the volume of loss of trabecular tissue was measured.
  • the periodontitis-induced group showed a statistically significant loss of cancellous bone volume compared to the normal group, confirming normal periodontitis induction, and periodontitis in the L. curvatus SMFM2016-NK treatment group. It was confirmed that the volume of cancellous bone loss was reduced by 11.6% compared to the induced group (FIG. 2).
  • Histopathological examination was performed after H&E staining using specimens of experimental animals measured by Micro-CT. Histomorphological changes were scored based on the following criteria, and inflammatory cell numbers, osteoblast cell numbers, osteoclast cell numbers, and alveolar process volumes were examined. Items were measured (Table 3).
  • the histomorphological change score was significantly reduced in the L. curvatus SMFM2016-NK treatment group after periodontitis induction compared to the periodontitis-inducing group, which was the L. curvatus of the present invention. This indicates that inflammatory cell infiltration in the gingival and periodontal ligaments was reduced, and alveolar bone resorption and cementum destruction were significantly reduced when the SMFM2016-NK strain was treated (FIG. 3).
  • curvatus SMFM2016-NK treatment group after periodontitis induction Periodontitis+ L. curvatus SMFM2016-NK+ statistically significant, compared to normal group by pairwise t-test ( p ⁇ 0.05)* statistically significant, compared to negative control group by pairwise t-test ( p ⁇ 0.05)
  • Example 6 Lactobacillus curvatus Characterization of SMFM2016-NK strain
  • the strain of the present invention was cultured in MRS broth (BD, USA) at 35° C. for 12 hours.
  • the bacteria were subcultured in MRS broth containing slide glass (0.5x0.5 cm) so that the bacteria grew attached to the slide glass surface.
  • a 1.8% glutaldehyde solution was dropped on a piece of glass slide and washed with sterile distilled water. After dropping a 2% osmium tetroxide (Sigma, St. Louis, MO, USA) solution, the reaction was carried out in the dark at room temperature.
  • HMDS hexamethyldisilazane
  • FE-SEM field emission scanning electrochemical microscopy
  • DNA link (Seoul) was requested to analyze the genome.
  • a 20 kb library was constructed using the Pacific Biosciences system (Pacific Biosciences, Menlo Park, CA, USA) sequencing platform, and de novo assembly was performed based on the HGAP.3 protocol.
  • the strain was identified as a strain consisting of a total of 4 contigs, and as a result of confirming each contig using the BLAST program based on the NCBI GenBank database, one chromosome (2,139,352 bp, GC content 41.97) %) and the genome with three plasmids.
  • the chromosome was confirmed to be most similar to L. curvatus.
  • the genetic characteristics (location of gene, coding region, function of gene) of the chromosome of the corresponding strain were analyzed, and structural characteristics ( The number of genes, their location) was confirmed with the following DNA plot. Specifically, it was confirmed that the L. curvatus SMFM2016-NK chromosome consists of 2,307 coding sequences (CDS), 18 rRNA, 67 tRNA, and 8,500 transcripts (FIG. 5).

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Abstract

La présente invention concerne une souche de Lactobacillus curvatus SMFM2016-NK (numéro d'enregistrement : KCTC14138BP), et une composition alimentaire pour la prévention ou le traitement de maladie buccale, une composition pharmaceutique pour la prévention ou le traitement de maladie buccale, des probiotiques, et une composition d'aliment pour animaux, comprenant chacune la souche. La souche de Lactobacillus curvatus SMFM2016-NK (numéro d'enregistrement : KCTC14138BP) selon la présente invention, qui est une souche probiotique nouvellement isolée et identifiée à partir de kimchi, présente l'effet de diminuer le nombre d'ostéoclastes, augmenter le nombre d'ostéoblastes, diminuer le volume de perte de l'os spongieux, augmenter le volume de l'os alvéolaire, et diminuer le nombre de cellules inflammatoires, et une excellente activité antibactérienne contre les bactéries nuisibles dans la bouche et, peut être ainsi utilisée de manière avantageuse dans une composition pour la prévention, le soulagement et le traitement d'une maladie buccale.
PCT/KR2020/007649 2020-03-23 2020-06-12 Probiotiques pour prévenir ou traiter la maladie d'alzheimer, et composition alimentaire les comprenant pour prévenir ou traiter une maladie buccale WO2021194011A1 (fr)

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KR102488905B1 (ko) * 2021-12-17 2023-01-18 주식회사 메디오젠 신규한 락토바실러스 커바투스 균주 및 이의 용도
KR102573674B1 (ko) * 2021-12-28 2023-09-01 한국식품연구원 라티락토바실러스 커바투스 WiKim0140를 포함하는 염증성 질환의 예방, 개선 또는 치료용 조성물
KR20240115991A (ko) * 2023-01-19 2024-07-29 재단법인 대구경북첨단의료산업진흥재단 골 질환 예방 또는 치료를 위한 신규 유산균 및 이의 배양액

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