[go: up one dir, main page]

WO2019103198A1 - Souche de lactobacillus fermentum kbl 375 et son utilisation - Google Patents

Souche de lactobacillus fermentum kbl 375 et son utilisation Download PDF

Info

Publication number
WO2019103198A1
WO2019103198A1 PCT/KR2017/013498 KR2017013498W WO2019103198A1 WO 2019103198 A1 WO2019103198 A1 WO 2019103198A1 KR 2017013498 W KR2017013498 W KR 2017013498W WO 2019103198 A1 WO2019103198 A1 WO 2019103198A1
Authority
WO
WIPO (PCT)
Prior art keywords
strain
lactobacillus
kbl
present
lactobacillus fermentum
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/KR2017/013498
Other languages
English (en)
Korean (ko)
Inventor
고광표
김운기
이준철
이진우
장유진
고우리
이기욱
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kobiolabs Inc
Original Assignee
Kobiolabs Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kobiolabs Inc filed Critical Kobiolabs Inc
Publication of WO2019103198A1 publication Critical patent/WO2019103198A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • 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
    • 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
    • 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
    • C12N1/205—Bacterial isolates
    • 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
    • A23V2200/32—Foods, ingredients or supplements having a functional effect on health having an effect on the health of the digestive tract
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/143—Fermentum
    • 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

  • the present invention relates to a lactobacillus fermentum KBL 375 strain and a use thereof, and more particularly to a lactobacillus fermentum KBL 375 strain which is a novel probiotic derived from a human gastrointestinal tract, a strain of the strain, a culture of the strain A composition for health functional food for improving intestinal health, and a pharmaceutical composition for the treatment or prevention of intestinal diseases, which contain an effective amount of at least one member selected from the group consisting of the above-mentioned disruption of said strain and the extract of said strain.
  • Probiotics refer to microorganisms having antimicrobial activity and enzymatic activity that aid intestinal microbial balance and products produced by the microorganisms.
  • probiotics are defined as live cells in the form of single or complex strains that are supplied to humans or animals in the form of dried cells or fermented products to improve intestinal microflora.
  • the characteristics that probiotics should possess are those that survive on the way to the human intestine as a habitat, non-pathogenic, non-toxic, and enteric.
  • they should maintain viability and activity prior to consumption in the transfer medium, be sensitive to antibiotics used to prevent infection, and not have plasmids resistant to antibiotics.
  • resistance to acids, enzymes and bile in the intestinal environment should be established.
  • probiotics include Bacillus sp . , Lactobacillus sp . which produces lactic acid, amylase, protease, lipase, cellulase and phosphatase, Photosynthetic bacteria that use substances (ammonia, hydrogen sulfide, amines, etc.) in the metabolic process to prevent odor induction are examples.
  • Bacillus and Lactobacillus species are known as very useful probiotics because of the existence of various strains producing various antimicrobial substances.
  • These lactic acid bacteria produce an antimicrobial peptide called bacteriocin, which has an antimicrobial mechanism that is not related to the antibiotic resistance mechanism.
  • Bacteriocin has polymorphic characteristics with remarkable differences in molecular weight, biochemical properties, antimicrobial range and mechanism to the host. Klaenhammer is defined as a protein or protein complex with direct antimicrobial activity against bacteriophage-producing bacteria and closely related species (Klaenhammer, TR. Biochimie 70: 337-379, 1998).
  • IBS is characterized by abdominal pain and / or discomfort associated with altered bowel movements or bowel habits, and these symptoms are not explained by structural or biochemical abnormalities.
  • the feeling of urgency, abdominal distension and incomplete bowel movement are also common symptoms of IBS.
  • the disease is classified as a functional gastrointestinal disorder including functional bloating, non-atrial chest pain, non-ulcer dyspepsia, and chronic constipation or diarrhea (Longstreth GH et al., 2006).
  • IBS has a profound effect on morbidity and quality of life, beyond abdominal pain and discomfort, since related symptoms act on both patient well-being and normal functional aspects (Dean BB et al., 2005).
  • WO 96/29083 and EP 554418 disclose Lactobacillus plantarum 299v (DSM 6595) and Lactobacillus casei ssp. Lactobacillus casei ssp . rhamnosus ) 271 (DSM 6594).
  • EP 415941 discloses a process for preparing a nutritional composition which comprises treating an oat gruel with an enzyme before mixing with lactobacilli.
  • U.S. Patent No. 7195906 discloses a Bifidobacterium strain isolated from a human gastrointestinal tract that has been resected and cleansed to treat inflammatory diseases, particularly gastrointestinal inflammatory actions such as IBD and IBS.
  • strains showing an excellent effect for improving intestinal health including irritable bowel syndrome have not yet been found, and researches for finding strains showing the above effects have continued at a number of research institutes.
  • the inventors of the present invention focused on the fact that the health promotion effect of probiotics is strain-specific rather than general characteristics of genus and species (Report of a joint FAO / WHO working group on drafting guidelines the evaluation of probiotics in food, London Ontario, Canada, 2002), by screening various strains of the human digestive tract and vagina derived was identified a novel strain outstanding have immunomodulatory effects, the intestinal health improvement of the strains in vitro and all in vivo, thereby completing the present invention.
  • the present invention provides a lactobacillus perfume KBL 375 (Accession No. KCTC 13381BP) strain.
  • the present invention also provides a composition comprising an effective amount of at least one member selected from the group consisting of a bacterium of Lactobacillus perfumant KBL 375 (Accession No. KCTC 13381BP), a culture of said strain, a lysate of said strain and an extract of said strain to provide.
  • a composition comprising an effective amount of at least one member selected from the group consisting of a bacterium of Lactobacillus perfumant KBL 375 (Accession No. KCTC 13381BP), a culture of said strain, a lysate of said strain and an extract of said strain to provide.
  • the present invention also provides a method for the treatment and / or prophylaxis of intestinal diseases comprising enterococci, which comprise an effective amount of at least one selected from the group consisting of strains of Lactobacillus perfumant KBL 375 (Accession No. KCTC 13381BP), the culture of the strain, the lysate of the strain, Or a pharmaceutically acceptable salt thereof.
  • enterococci which comprise an effective amount of at least one selected from the group consisting of strains of Lactobacillus perfumant KBL 375 (Accession No. KCTC 13381BP), the culture of the strain, the lysate of the strain, Or a pharmaceutically acceptable salt thereof.
  • FIG. 2 shows the result of confirming the immunoregulatory function of Lactobacillus fermentum KBL 375 strain according to the present invention using PBMC.
  • FIG. 3 shows the results of comparing the immunoregulatory function of the Lactobacillus perfumant KBL 375 strain according to the present invention with that of the other lactobacillus fermentum strains, KCTC 5467 strain and RAW 264.7 cell line.
  • FIG. 4 shows the result of comparing the tight junction synergistic effect of the Lactobacillus perfumant KBL 375 strain of the present invention with that of other Lactobacillus fermentum strains, SNUV 417 strain and Caco-2 cell line.
  • FIG. 5 shows the result of administering Lactobacillus perfumant KBL 375 strain of the present invention to enteritis-induced mouse models and observing changes in body weight and colon length.
  • Lactobacillus perfumant KBL 375 strain of the present invention is a graph comparing the growth rate of Lactobacillus perfumant KBL 375 strain of the present invention with that of Lactobacillus fermentum KCTC5049 and Lactobacillus fermentum KCTC5467, which are different Lactobacillus fermentum strains, from De Man, Rogosa and Sharpe (MRS) This is a result.
  • Lactobacillus perfumant KBL 375 (Accession No. KCTC 13381BP) showed particularly excellent effects on immune regulation
  • KCTC 13381BP Lactobacillus perfumant KBL 375
  • the present invention relates to a strain of lactobacillus fermentum KBL 375 (Accession No. KCTC 13381BP) which is a novel probiotic in a viewpoint, characterized in that the strain comprises the 16s rDNA sequence shown in SEQ ID NO: 1 do.
  • Lactobacillus perfume KBL 375 strain had immunosuppressive effect as well as intimate tightening effect of intestinal wall, and when lactobacillus fermentum KBL 375 strain was administered to an enteritis-induced animal model, Weight loss and colon length reduction were significantly improved.
  • the present invention relates to a method for the production of an effective amount of at least one compound selected from the group consisting of Lactobacillus perfume KBL 375 (Accession No. KCTC 13381BP) strain, a culture of the strain, a lysate of the strain and an extract of the strain ≪ / RTI >
  • the composition may be a food or food additive composition, but the present invention is not limited thereto.
  • foods that are effective for improving intestinal health or preventing intestinal diseases for example, main ingredients, additives, food additives, Or as a functional beverage.
  • &quot means a natural product or a processed product containing one or more nutrients, preferably a state of being able to be eaten directly through a certain degree of processing, Additives, health functional foods and functional beverages.
  • Foods to which the food composition according to the present invention can be added include, for example, various foods, beverages, gums, tea, vitamin complexes, and functional foods.
  • the food may include special nutritive foods (e.g., crude oil, spirits, baby food, etc.), meat products, fish meat products, tofu, mackerel, noodles (Such as soy sauce, soybean paste, kochujang, mixed potatoes), sauces, confectionery (eg, snacks), candies, chocolate, gums, ice cream, milk products (eg, fermented milk, cheese, But are not limited to, pickled foods (various kinds of kimchi, pickles, etc.), beverages (e.g., fruit drinks, vegetable beverages, beverages, fermented beverages and the like) and natural seasonings (e.g.
  • the food, beverage or food additive may be prepared by a conventional production method.
  • the above-mentioned health functional food refers to a food group to which the function of the food is added to a specific purpose by physical, biochemical, and biotechnological techniques, and to control the bio-defense rhythm of the food composition, Quot; means a food which is processed by being designed so as to sufficiently express the body controlling function related to the living body.
  • the functional food may include a food-acceptable food-aid additive, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of functional foods.
  • the functional beverage refers to a generic term for eliminating thirst or for enjoying a taste, and includes a composition for improving or preventing symptoms of intestinal diseases as an essential ingredient at a specified ratio, And may contain various flavoring agents or natural carbohydrates as additional ingredients such as ordinary beverages.
  • the food containing the composition for food for improving or preventing symptoms of intestinal diseases according to the present invention may contain flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, And carbonating agents used in fillers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, And these components can be used independently or in combination.
  • flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, And carbonating agents used in fillers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, And these components can be used independently or in combination.
  • the amount of the composition according to the present invention may be 0.001% by weight to 100% by weight, preferably 1% by weight to 99% by weight, And may be contained in a ratio of 0.001 g to 10 g, preferably 0.01 g to 1 g, based on 100 ml in the case of beverages.
  • 0.001 g to 10 g preferably 0.01 g to 1 g, based on 100 ml in the case of beverages.
  • the active ingredient has no problem in terms of safety, it can be used in an amount of more than the above range, so it is not limited to the above range.
  • the food composition of the present invention may be prepared in the form of a composition suitable for human or animal ingestion, or the Lactobacillus perfume KBL 375 strain may be added to an independent or acceptable carrier. That is, it can be added to foods that do not contain other probiotic bacteria and foods that already contain some probiotic bacteria.
  • other microorganisms that can be used in combination with the strain of the present invention are suitable for human or animal ingestion and can inhibit pathogenic harmful bacteria upon ingestion or improve the microorganism balance in mammalian gut Those having probiotic activity, and are not particularly limited.
  • probiotic microorganism Saccharomyces in MRS (Saccharomyces), Candida (Candida), blood teeth (Pichia) and sat rulrop cis yeast (yeast), Aspergillus (Aspergillus) containing (Torulopsis), Rhizopus Perth ( Rhizopus), myuko (Mucor), penicillin (Penicillium) fungi and Lactobacillus bacteria (Lactobacillus), Bifidobacterium (Bifidobacterium), current Kono Stock (Leuconostoc), Lactococcus (Lactococcus), Bacillus (Bacillus), such as, Streptococcus Bacteria belonging to the genera Streptococcus , Propionibacterium , Enterococcus , Pediococcus , and the like.
  • probiotic microorganisms include, but are not limited to, Saccharomyces cerevisiae , Bacillus coagulans , Bacillus licheniformis , Bacillus subtilis , Bifidobacterium bifidum), Bifidobacterium Infante Tees (Bifidobacterium infantis), Bifidobacterium rongeom (Bifidobacterium longum), Enterococcus passive help (Enterococcus faecium), Enterococcus faecalis (Enterococcus faecalis), Lactobacillus ash also filler's (Lactobacillus acidophilus, Lactobacillus alimentarius, Lactobacillus casei , Lactobacillus curvatus , Lactobacillus delbruckii , Lactobacillus johnsonii , Lactobacillus spp ., Lactobacillus farc
  • the effect of the present invention can further be enhanced by additionally incorporating a probiotic microorganism mixed bacterium having an excellent probiotic activity and an excellent immunity activity and anticancer activity in the food composition of the present invention.
  • a probiotic microorganism mixed bacterium having an excellent probiotic activity and an excellent immunity activity and anticancer activity
  • carriers which can be used in the food composition of the present invention include extender, high fiber additive, encapsulating agent, lipid and the like, and examples of such carriers are well known in the art.
  • the Lactobacillus perfume KBL 375 of the present invention may be in lyophilized or encapsulated form or in the form of a culture suspension or a dry powder.
  • composition of the present invention may also be provided in the form of an animal feed additive containing the strain or animal feed containing it.
  • the animal feed additive of the present invention may be in the form of a dry or liquid preparation, and may further contain other non-pathogenic microorganisms other than Lactobacillus perfume KBL 375.
  • microorganisms which can be added include lactic acid bacteria such as Bacillus subtilis capable of producing protease, lipolytic enzyme and glycosyltransferase, lactose with physiological activity and organic decomposition in anaerobic conditions such as bovine Lactobacillus strains, such as Aspergillus oryzae (Slyter, LLJ Animal Sci. 1976, 43), which increase the body weight of livestock and increase milk yield and increase digestion and uptake of feed.
  • yeasts such as Saccharomyces cerevisiae (Johnson, D. E et al. J. Anim. Sci., 1983, 56, 735-739; Williams, PEV et al, 1990, 211) may be used.
  • the animal feed additive of the present invention may further comprise at least one enzyme preparation in addition to the above-mentioned Lactobacillus perfume KBL 375.
  • the added enzyme preparation can be either dry or liquid.
  • Enzyme agents include lipase such as lipase, phytase which decomposes phytic acid to make phosphate and inositol phosphate, Amylase which is an enzyme which hydrolyzes an ⁇ -1,4-glycoside bond contained in glycogen and the like, phosphatase which is an enzyme which hydrolyzes an organic phosphate ester, cellulose which is an enzyme which hydrolyzes an organic phosphate ester, Carboxymethylcellulase which degrades xylose, xylase which degrades xylose, maltase which hydrolyzes maltose into glucose with glucose and maltase which hydrolyzes maltose into glucose Sugar-producing enzymes such as invertase and the like which
  • the lactobacillus fermentum KBL 375 strain of the present invention can be used as an animal feed additive in various feedstuffs such as peanut, pea, beet, pulp, grain by-products, animal powder, fish meal, And they can be used without limitation, either unprocessed or processed.
  • the processing is not necessarily limited to this.
  • a process in which a feed material is filled with a constant outlet under pressure in the state of being filled with feed, and in the case of a protein, extrusion is used in which denaturation and availability are increased .
  • Extrusion is advantageous in that it modifies the protein through heat treatment and destroys the antioxidant factor.
  • soy protein it is possible to improve protein digestibility through extrusion and to inactivate antitrophic factors such as trypsin inhibitor, which is one of protease inhibitors present in soybean, And the nutritional value of soy protein can be increased.
  • the present invention provides an effective amount of at least one selected from the group consisting of Lactobacillus fermentum KBL 375 (Deposit No. KCTC 13381BP) strain, a culture of the strain, a lysate of the strain and an extract of the strain And to a pharmaceutical composition for the treatment or prevention of intestinal diseases.
  • the pharmaceutical composition of the present invention may be provided as a composition that can be combined with cells of a live cell, a form of a dry strain, a culture of the strain, a lysate of the strain, or a pharmaceutically acceptable carrier or medium thereof.
  • the carrier or medium employed may be selected from the group consisting of a solvent, a dispersing agent, a coating, an absorption promoting agent, a controlled release agent (i.
  • the tablet formulations of the disclosed compositions may be coated by standard aqueous or non-aqueous techniques.
  • excipients, fillers, disintegrants, lubricants, antimicrobial agents, and coatings include, but are not limited to, pharmaceutically acceptable carriers and excipients for use as pharmaceutically acceptable inert carriers and examples of such additional ingredients It is not.
  • the enteric disease is selected from the group consisting of abdominal distension, abdominal discomfort, infectious diarrhea caused by pathogenic microorganisms, gastroenteritis, inflammatory bowel disease, neurodegenerative inflammatory syndrome, irritable bowel syndrome, small intestinal microorganisms, And includes diseases caused by normal barrier function impairment of the intestines.
  • IBD Inflammatory Bowel Disease
  • Ulcerative colitis affects only the large intestine. Inflammation and ulcers in ulcerative colitis are limited to the mucosal and submucosal layers, the two innermost layers of the four layers of the colon. In Crohn's disease, inflammation and ulcers can spread through all layers of the barrier in both the small intestine and the large intestine.
  • IBS Irritable Bowel Syndrome
  • the therapeutic or therapeutic treatment of intestinal disorders in mammals thus may include one or more of the following:
  • composition for the prevention or treatment of intestinal diseases may comprise a pharmaceutically effective amount of Lactobacillus perfumant KBL 375 (Accession No. KCTC 13381BP) alone or in combination with one or more pharmaceutically acceptable carriers, excipients or diluents .
  • the term "effective amount (or an effective amount)" means an amount which is sufficiently high to deliver a desired effect but which is sufficiently small to sufficiently prevent serious side effects within the medical judgment range.
  • the amount of the microorganism to be administered into the body by the composition of the present invention can be appropriately adjusted in consideration of the route of administration and the subject to be administered.
  • the composition of the present invention may be administered to the subject once or more daily.
  • Unit dose refers to units physically discrete suitable for unit administration for human subjects and other mammals and each unit contains a suitable pharmaceutical carrier and contains a predetermined amount of the microorganism of the invention which exhibits a therapeutic effect .
  • the dosage unit for oral administration to an adult patient preferably contains 0.001 g or more of the microorganism of the present invention, and the oral dose of the composition of the present invention is 0.001 to 10 g, preferably 0.01 to 5 g, at one time.
  • the pharmaceutically effective amount of the microorganism of the present invention is 0.01 to 10 g / day.
  • the dosage varies depending on the severity of the enteric disease of the patient and the microorganism and auxiliary active ingredient used.
  • the total daily dose may be divided into several doses and administered sequentially as necessary. Thus, the dosage ranges do not limit the scope of the invention in any way.
  • pharmaceutically acceptable refers to a composition that is physiologically acceptable and does not normally cause an allergic reaction such as gastrointestinal disorder, dizziness, or the like when administered to humans.
  • compositions of the present invention may be formulated using methods known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal.
  • the formulations may be in the form of powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatine capsules, sterile injectable solutions, sterile powders.
  • composition for preventing or treating intestinal diseases according to the present invention may be administered through various routes including oral, transdermal, subcutaneous, intravenous, or muscular, and the dose of the active ingredient may vary depending on the administration route,
  • the composition for preventing or treating gastrointestinal diseases according to the present invention may be appropriately selected according to various factors such as body weight and the severity of the patient and the composition for preventing or treating gastrointestinal diseases according to the present invention may be used in combination with a known compound having an effect of preventing, Lt; / RTI >
  • enteric coating includes all known pharmaceutically acceptable coatings which are not degraded by stomach acid to maintain the coating, but which are sufficiently degraded in the small intestine so that the active ingredient can be released into the small intestine do.
  • enteric coating " of the present invention remains intact for at least 2 hours when contacting an artificial juice, such as a HCl solution at pH 1, at 36 ° C to 38 ° C, and is preferably subsequently maintained at a pH of 6.8, such as KH 2 PO 4 buffer Refers to a coating that decomposes within 30 minutes of the juice.
  • the enteric coating of the present invention is coated on one core in an amount of about 16 to 30, preferably 16 to 20 or 25 mg or less.
  • the thickness of the enteric coating of the present invention is 5 to 100 mu m, preferably 20 to 80 mu m, satisfactory results are obtained as an enteric coating.
  • Materials for enteric coatings are appropriately selected from known polymeric materials. Suitable polymeric materials are described in numerous publications (L. Lachman et al., The Theory and Practice of Industrial Pharmacy, 3rd ed., 1986, pp. 365-373; H. Sucker et al., Pharmazeutician Technologie, Thieme, 1991, pp. 355-359 Hager Handbuchter pharmazeuticiantechnik, 4th Edition, Vol.7, pp.
  • the enteric coating of the present invention can be prepared using conventional enteric coating methods in which an enteric coating solution is sprayed onto the core.
  • Suitable solvents for use in enteric coating processes include alcohols such as ethanol, ketones such as acetone, halogenated hydrocarbon solvents such as dichloromethane (CH2Cl2), and mixed solvents of these solvents may also be used.
  • a softener such as di (di) n-butyl phthalate or triacetin is added to the coating solution in a ratio of from about 0.05 to about 0.3 (coating material to softener) per coat. It is appropriate to carry out the spraying process continuously and it is possible to adjust the spraying amount in consideration of the conditions of the coating.
  • the spraying pressure can be varied and generally satisfactory results are obtained with spray pressures of from about 1 to about 1.5 bar.
  • the present invention provides the use of the strain or composition for use in the prevention or treatment of intestinal diseases and the use of the strain or composition for the preparation of a therapeutic agent for intestinal diseases.
  • the term " prevention " relates to averting, delaying, impeding or hindering the disease.
  • the term " treatment " relates to caring for a subject afflicted with a disease in order to ameliorate, cure, or reduce the symptoms of the disease or to reduce or stop the progression of the disease.
  • the present invention provides a method for preventing or treating enteric diseases, comprising the step of administering a pharmaceutically effective amount of the above strain or composition to a subject in need of prevention or treatment of bowel disease or improvement of intestinal health.
  • the individual to which the composition for preventing or treating bowel disease can be administered includes all animals.
  • it may be an animal such as a dog, a cat, or a mouse.
  • a THP-1 cell line, a leukemic mononuclear cell, and a peripheral blood mononuclear cell (PBMC) were screened for a probiotic strain having an immunoregulatory function.
  • a strain derived from a human gut or vagina was added to each of the above two cell lines to a ratio of 1: 100, and inflammation of the cytokine IL-6, which is a main index of the inflammatory response, (IL-10 / IL-6) of the cytokine IL-10, which is the main indicator of the regulation of IL-10.
  • Lactobacillus gasseri For screening, two kinds of Lactobacillus gasseri , one Lactobacillus reuteri , five Lactobacillus rhamnosus , two Lactobacillus fermentum , and two Lactobacillus paracase , (Lactobacillus paracasei) 4 species, Lactobacillus salivarius (Lactobacillus salivarius) 4 species, Lactobacillus Planta Room (Lactobacilus plantarum ), two Lactobacillus acidophilus strains and two Lactococcus lactis strains were used as a total of 23 strains
  • Lactobacillus fermentum KBL 375 (KCTC 13381BP) showed an IL-10 / IL-6 value of 4.94 in the THP-1 cell line and an IL-10 / IL-6 value of 7.79 in the peripheral blood mononuclear cell Effect.
  • Table 1 shows the results of comparing IL-10 / IL-6 values between two Lactobacillus fermentum strains among the 23 probiotic strains used in the present screening.
  • Lactobacillus fermentum KBL 375 strain showed a very high IL-10 / IL-6 value, and the strain was predicted to be a highly immunomodulatory strain. Lactobacillus fermentum KBL 375 And further experiments were conducted using strains.
  • the THP-1 cell line was dispensed into each well of a 24-well plate (1 ⁇ 10 5 cells) and matured into macrophages. Lactobacillus fermentum KBL 375 strain was added 3 hours after the culture medium was replaced Each well was dispensed at a rate of 1 ⁇ 10 7 cells on the basis of the viable cell count.
  • LPS was treated at a concentration of 1 ⁇ g / ml instead of Lactobacillus fermentum KBL 375 strain under the same conditions to induce an inflammatory reaction. After 24 hours, the amount of each cytokine was measured according to the manufacturer's method using a BD cytometric Bead Array (CBA) human inflammation kit (Cat No. 551811).
  • CBA BD cytometric Bead Array
  • PBMC cells were dispensed into each well of a 24-well plate at 5 ⁇ 10 5 cells, and then Lactobacillus perfumant KBL 375 was added at a concentration of 5 ⁇ 10 6 cells per viable cell count.
  • Escherichia coli O157 (H7 EC4115) isolated from the intestine was added to the wells at a concentration of 5 ⁇ 10 6 cells per well or LPS was added at a concentration of 500 ng / ml.
  • the PBMC cell group prepared under the above conditions was cultured for 5 days, and then the cells were collected.
  • the synthesized cDNA was subjected to real-time PCR using a Rotor-Gene® Q (Qiagen) kit and a Rotor-Gene SYBR Green PCR kit (Qiagen) bet, Th2 marker gene, GATA3, Th17 marker gene RORrt, and Immune regulatory T cell (Treg cell) marker gene, FOXP3 mRNA expression.
  • sequences of the primers for confirming each gene expression were forward 5'-CCC CAA GGA ATT GAC AGT TG-3 '(reverse) 5'-GGG AAA CTA AAG CTC ACA AAC-3' , 5'-CTG CAA TGC CTG TGG GCT C-3 '(reverse) 5'-GAC TGC AGG GAC TCT CGC T-3' for amplification of GATA3, 5'- AAG ACT CAT CGC CAA AGC AT-3 '(reverse) 5'-TCC ACA TGC TGG CTA CAC A-3', 5'-TCA AGC ACT GCC AGG CG-3 'reverse for FOXP3 amplification 5'-CAG GAG CCC TTG TCG GAT-3 'was used.
  • the Lactobacillus perfume KBL375 strain significantly decreased the expression levels of T-bet, GATA3 and RORrt genes, which are effector cell markers of T cells, compared with E. coli, while Foxp3 mRNA was significantly increased.
  • RAW 264.7 cell lines were each subdivided into each well of a 24-well plate at a rate of 1 ⁇ 10 5 cells.
  • LPS was treated at a concentration of 2 ng / ml to induce an inflammatory reaction
  • KCTC 5467 a representative subspecies of Lactobacillus fermentum Or Lactobacillus fermentum KBL 375 was added to each well at a rate of 1 ⁇ 10 6 cells per well and then cultured for 24 hours to determine the concentration of TNF and IL-10 in an ELISA kit (Mouse TNF ELISA Set II No. 558534, BD OptEIATM), and Mouse IL-10 ELISA Set (Cat No. 555252, OptEIATM).
  • the transepithelial / transendothelial electrical resistance (TEER) assay was performed to confirm the effect of Lactobacillus fermentum KBL 375 (KCTC 13381BP) on the intestinal wall adhesion tightening effect.
  • Lactobacillus fermentum SNUV 417 strain derived from human vagina was used as a comparative experimental group.
  • the Caco-2 (ATCC® HTB-37 TM) cell line for the TEER assay contains 20% FBS, 1% non-essential amino acids solution, 1% HEPES, 1.5% sodium bicarbonate solution, 50 ⁇ g / ml gentamicin, penicillin-streptomycin at 37 ° C and 5% CO 2 .
  • Caco-2 cells were dispensed at a rate of 3 ⁇ 10 4 cells / well in a 12-well plate, and the medium was changed every 2 days. Then, the medium was changed to a medium without FBS and antibiotics on the 6th day. On the 7th day, the plate was kept at room temperature for at least 20 minutes. The temperature of the plate was adjusted to room temperature, and the TEER was measured using a VOM resistance meter (World Precision Instruments, Sarasota, FL, USA). Then, the Lactobacillus fermentum KBL 375 strain of the present invention and the Lactobacillus fermentum SNUV 417 strain of the comparative experiment group were all washed with PBS and then released into MEM.
  • FACS FACS was used to perform live / dead cell counting and the strain was treated with Caco-2 cells so as to have a cell count of 3 ⁇ 10 7 cells / well.
  • Lactobacillus fermentum KBL 375 strain As a result, as shown in FIG. 4, it was confirmed that the cells treated with Lactobacillus fermentum KBL 375 strain were significantly increased compared with the control and comparative experimental group, Lactobacillus perfumant SNUV 417 strain. This indicates that the Lactobacillus perfume KBL 375 strain has an effect of enhancing tight junctions in the intestinal wall and can be used for the treatment of irritable bowel syndrome.
  • Lactobacillus fermentum KBL 375 (KCTC 13381BP) strain is in vivo And the effect of improving the bowel function was also confirmed.
  • C57BL / 6 mice were divided into 10 clusters of each, and 2% DSS was dissolved in tap water, and drinking water for 9 days to induce enteritis.
  • 200 ⁇ l of PBS was orally administered to the control mice daily
  • Lactobacillus fermentum KBL 375 strain was diluted to 2 ⁇ 10 10 CFU / ml in the test mice and then orally administered in 200 ⁇ l each day. Thereafter, body weight changes of control and experimental mice were measured daily for 9 days during which DSS was induced and enteritis was induced. On the 9th day after feeding DSS, mice were autopsied and colon length was measured.
  • Example 5 Productivity Assay of Lactobacillus fermentum KBL 375 strain
  • Lactobacillus fermentum KBL 375 (KCTC 13381BP)
  • two representative subspecies L. fermentum KCTC5049, L. fermentum KCTC5467
  • Lactobacillus fermentum were registered in the BRC.
  • FIG. 6 when Lactobacillus fermentum strains were cultured in De Man, Rogosa and Sharpe (MRS) medium, respectively, Lactobacillus perfumant KBL 375 strain had a faster growth rate than the other two Lactobacillus fermentum strains I could confirm.
  • the specific growth rate of Lactobacillus fermentum KBL375 was 0.756, which was 1.1 to 1.3 times faster than that of other Lactobacillus fermentum (Table 2).
  • Lactobacillus fermentum KBL 375 and Lactobacillus fermentum KCTC5467 were required to have a doubling time of 0.91 hours, 1.16 hours and 1.02 hours, respectively. (Table 2). Therefore, it was confirmed that Lactobacillus fermentum KBL 375 was superior to other Lactobacillus fermentum strains reported above by more than 10%.
  • Lactobacillus fermentum KBL375 Lactobacillus fermentum KCTC5049 and Lactobacillus fermentum KCTC5467
  • Strains Specific growth rate (h -1 ) Doubling time (h) Character L. fermentum KBL375 0.756 0.916 Isolated from feces adult L. fermentum KCTC5049 0.677 1.163 Isolated from saliva L. fermentum KCTC5467 0.596 1.024 Isolated from intestine
  • lactobacillus fermentum KBL 375 strain according to the present invention is excellent in immunomodulating function, excellent in reinforcing effect on intestinal wall adhesion, and is effective in treating enteritis by inhibiting weight loss and reduction of colon length due to enteritis , And as a probiotic material for improving intestinal health function.
  • Lactobacillus fermentum KBL 375 strain according to the present invention has an advantage of being industrially advantageous because of its excellent productivity because of its rapid growth rate compared with similar strains during culturing.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Food Science & Technology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Nutrition Science (AREA)
  • Polymers & Plastics (AREA)
  • Molecular Biology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

La présente invention concerne une souche de Lactobacillus fermentum KBL 375, qui est une nouvelle souche probiotique issue du tractus digestif humain, et une utilisation de la souche pour l'amélioration de la santé intestinale et le traitement ou la prévention de l'entéropathie. La souche de Lactobacillus fermentum KBL 375 selon la présente invention présente une excellente activité immunorégulatrice et un effet d'amélioration remarquable sur les jonctions serrées de la paroi intestinale et supprime la perte de poids et la diminution de la longueur du côlon induites par l'entérite pour exercer un effet thérapeutique sur l'entérite et peut être utilisée utilement en tant que matériau probiotique pour améliorer les fonctions associées à la santé intestinale.
PCT/KR2017/013498 2017-11-24 2017-11-24 Souche de lactobacillus fermentum kbl 375 et son utilisation Ceased WO2019103198A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020170158315A KR102024883B1 (ko) 2017-11-24 2017-11-24 락토바실러스 퍼멘텀 kbl 375 균주 및 그 용도
KR10-2017-0158315 2017-11-24

Publications (1)

Publication Number Publication Date
WO2019103198A1 true WO2019103198A1 (fr) 2019-05-31

Family

ID=66630636

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2017/013498 Ceased WO2019103198A1 (fr) 2017-11-24 2017-11-24 Souche de lactobacillus fermentum kbl 375 et son utilisation

Country Status (2)

Country Link
KR (1) KR102024883B1 (fr)
WO (1) WO2019103198A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3818985A1 (fr) 2019-11-06 2021-05-12 Sanprobi Spolka Z Ograniczona Odpowiedzialnoscia Spolka Lactobacillus fermentum pl9 et son application
CN116004455A (zh) * 2022-12-28 2023-04-25 广西爱生生命科技有限公司 一株具有分泌5-羟色胺功能的发酵乳杆菌菌株a21196及其应用
EP4332214A4 (fr) * 2021-04-27 2024-11-06 Atogen Co., Ltd. Nouvelle souche de lactobacillus fermentum atg-v5, ou composition pour renforcer l'immunité la comprenant

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115135750B (zh) * 2019-12-31 2024-12-24 Gi生物群系公司 发酵乳杆菌菌株及含有其的用于预防或治疗代谢疾病的组合物
KR102273157B1 (ko) * 2020-07-20 2021-07-05 (주)비알컴퍼니 소화 촉진 및 혈당 상승 억제 효과를 갖는 유기농 밀면 및 이의 제조방법
WO2022124871A1 (fr) * 2020-12-11 2022-06-16 주식회사 바이오뱅크힐링 Souche de lactobacillus sp. , souche d'eubacterium sp., ou souche de coprococcus sp., vésicules dérivées de ces dernières, et utilisation anti-inflammatoire et antibactérienne correspondante
KR102296287B1 (ko) * 2020-12-11 2021-09-01 주식회사 바이오뱅크힐링 락토바실러스 퍼멘텀 균주, 및 그의 유래의 소포체 및 그의 항염증 및 항균 용도
KR20230026116A (ko) * 2021-08-17 2023-02-24 순천향대학교 산학협력단 락토바실러스 퍼멘툼 균주 및 이를 포함하는 장염 예방 또는 치료용 약학적 조성물
KR20240046081A (ko) * 2022-09-30 2024-04-08 주식회사 에이치이엠파마 락토바실러스 퍼멘텀 hem1036 균주를 유효성분으로 포함하는 암 치료 부작용 개선용 조성물
KR20250152751A (ko) * 2024-04-16 2025-10-24 주식회사 위바이옴 락토바실러스 퍼멘텀 균주의 면역 증진 용도

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20030064030A (ko) * 2002-01-25 2003-07-31 주식회사 푸코 위액 및 담즙산에 대한 내성이 높아 장내 생존력이 우수한한국인 분변 유래 유용 프로바이오틱 생균 및 그의 용도
US20070134220A1 (en) * 2003-04-02 2007-06-14 Axcan Pharma S.A. Lactobacillus fermentum strain and uses thereof
KR101452234B1 (ko) * 2013-07-22 2014-10-22 (주) 피엘바이오 장수마을 성인으로부터 분리한 배변활동에 도움을 주는 신규한 유산균 락토바실러스 퍼멘텀
EP2707476B1 (fr) * 2011-05-06 2015-07-08 Biotmicrogen, S.L. Souche de lactobacillus fermentum cect 7472 ayant des propriétés probiotiques
WO2017047968A1 (fr) * 2015-09-15 2017-03-23 경희대학교 산학협력단 Nouveau lactobacillus présentant diverses fonctions et son utilisation

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SE463796B (sv) 1988-03-09 1991-01-28 Carl Erik Albertsson Foerfarande foer framstaellning av en naeringskomposition och daervid framstaelld naeringskomposition
SE469875C (sv) 1991-07-25 1997-04-14 Probi Ab Stam av tarmkoloniserande Lactobacillus samt komposition för profylax eller behandling av infektioner i magtarmkanalen
SE9501056D0 (sv) 1995-03-23 1995-03-23 Probi Ab Epithelial adherent lactobacilli
ID29150A (id) 1999-01-15 2001-08-02 Entpr Ireland Cs Penggunaan lactobacillus salivarius

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20030064030A (ko) * 2002-01-25 2003-07-31 주식회사 푸코 위액 및 담즙산에 대한 내성이 높아 장내 생존력이 우수한한국인 분변 유래 유용 프로바이오틱 생균 및 그의 용도
US20070134220A1 (en) * 2003-04-02 2007-06-14 Axcan Pharma S.A. Lactobacillus fermentum strain and uses thereof
EP2707476B1 (fr) * 2011-05-06 2015-07-08 Biotmicrogen, S.L. Souche de lactobacillus fermentum cect 7472 ayant des propriétés probiotiques
KR101452234B1 (ko) * 2013-07-22 2014-10-22 (주) 피엘바이오 장수마을 성인으로부터 분리한 배변활동에 도움을 주는 신규한 유산균 락토바실러스 퍼멘텀
WO2017047968A1 (fr) * 2015-09-15 2017-03-23 경희대학교 산학협력단 Nouveau lactobacillus présentant diverses fonctions et son utilisation

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
DATABASE Nucleotide 27 April 2008 (2008-04-27), "Lactobacillus sp. KLDS 1.0710 16S Ribosomal RNA Gene , Partial Sequence", XP055618796, retrieved from NCBI Database accession no. EU600914.1 *
PARK, JONG-SU ET AL.: "Characterizatoin of Lactobacillus Fermentum PL 9988 Isolated from Healthy Elderly Korean in a Longevity Village", J. MICROBIOL. BIOTECHNOL., vol. 25, no. 9, 2015, pages 1510 - 1518, XP055618794 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3818985A1 (fr) 2019-11-06 2021-05-12 Sanprobi Spolka Z Ograniczona Odpowiedzialnoscia Spolka Lactobacillus fermentum pl9 et son application
EP4332214A4 (fr) * 2021-04-27 2024-11-06 Atogen Co., Ltd. Nouvelle souche de lactobacillus fermentum atg-v5, ou composition pour renforcer l'immunité la comprenant
CN116004455A (zh) * 2022-12-28 2023-04-25 广西爱生生命科技有限公司 一株具有分泌5-羟色胺功能的发酵乳杆菌菌株a21196及其应用

Also Published As

Publication number Publication date
KR102024883B1 (ko) 2019-09-24
KR20190060271A (ko) 2019-06-03

Similar Documents

Publication Publication Date Title
US11992511B2 (en) Lactobacillus paracasei strain and use thereof
WO2019103198A1 (fr) Souche de lactobacillus fermentum kbl 375 et son utilisation
JP5081242B2 (ja) 人間母乳から分離したプロバイオティック活性、及び体重増加抑制活性を有する乳酸菌
US10597740B2 (en) Bifidobacterium longum CBT BG7 strain for promotion of growth and nutraceutical composition for promotion of growth containing the same
WO2016186243A1 (fr) Nouvelle souche de bifidobacterium et composition alimentaire fonctionnelle la contenant pour stimuler la croissance
KR20240102112A (ko) 대장염 예방 또는 치료활성 및 장내 유해균에 항균활성을 지닌 락토바실러스 및 그의 용도
JP2008054556A (ja) 免疫賦活作用及び／又はアレルギー抑制作用を有し、且つ胃液耐性を有する新規乳酸菌
WO2023058801A1 (fr) Composition pour soulager, prévenir ou traiter un trouble intestinal, comprenant une souche de lactobacillus acidophilus kbl402 ou kbl409
RU2790297C2 (ru) Штамм lactobacillus paracasei и его применение
RU2790297C9 (ru) Штамм lactobacillus paracasei и его применение
HK40040941A (en) Lactobacillus paracasei strain and use thereof
BR122024010958A2 (pt) Cepa de lactobacillus paracasei kbl385, composições alimentar, de aditivo alimentar e farmacêuticas, bem como usos de uma composição compreendendo a cepa para prevenir ou tratar doenças intestinais, doenças alérgicas, doenças autoimunes ou doenças inflamatórias

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17933123

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 17933123

Country of ref document: EP

Kind code of ref document: A1