WO2023075357A1 - Souche de lactobacillus crispatus et composition pour la prévention ou le traitement de vaginite la comprenant - Google Patents

Souche de lactobacillus crispatus et composition pour la prévention ou le traitement de vaginite la comprenant Download PDF

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WO2023075357A1
WO2023075357A1 PCT/KR2022/016340 KR2022016340W WO2023075357A1 WO 2023075357 A1 WO2023075357 A1 WO 2023075357A1 KR 2022016340 W KR2022016340 W KR 2022016340W WO 2023075357 A1 WO2023075357 A1 WO 2023075357A1
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lactobacillus crispatus
strain
vaginitis
strains
culture
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PCT/KR2022/016340
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English (en)
Korean (ko)
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이성희
김영후
박재완
은수현
이동훈
전우진
한치영
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일동제약(주)
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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
    • C12N1/205—Bacterial isolates
    • 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
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A61P15/02—Drugs for genital or sexual disorders; Contraceptives for disorders of the vagina
    • 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
    • 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
    • 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
    • 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 novel Lactobacillus genus strain, a culture thereof, and a composition for treating vaginitis containing the same, and specifically, a novel Lactobacillus crispatus strain isolated from the vagina of a healthy woman, a culture thereof, It relates to a pharmaceutical composition and a health functional food composition for the prevention or treatment of vaginitis comprising.
  • vagina Under normal conditions, a woman's vagina is a slightly acidic environment due to many lactic acid bacteria inside the vagina, which plays a role in inhibiting the proliferation of pathogenic and harmful microorganisms.
  • the slightly acidic environment in the vagina is caused by external factors such as excessive vaginal washing, long-term administration of antibiotics or contraceptives, disruption of hormone balance due to pregnancy, childbirth, menopause, etc. If changes occur, it can be broken, which is known to cause vaginitis that causes discharge, itching, and pain.
  • Vaginitis is one of the common female gynecological diseases and is often accompanied by symptoms such as itching, burning, unpleasant odor and abnormal vaginal discharge.
  • BV bacterial vaginosis
  • VVC vulvovaginal candidiasis
  • Trichomoniasis 15 -20%)
  • Chlamydia or gonorrhea mainly causes cervicitis and is a major causative agent of pelvic inflammatory disease, and vaginal secretion may increase due to an increase in purulent secretion.
  • Bacterial vaginosis or Trichomonas vaginitis increase the risk of premature amniotic membrane rupture and preterm labor obstetrically, and are associated with an increase in postoperative complications such as infections. appropriate treatment is essential.
  • Drugs mainly used in the treatment of vaginitis include sulfonamides, antibiotics, disinfectants by chemical synthesis, etc., but the use of these drugs is fatal not only to harmful bacteria but also to beneficial lactic acid bacteria.
  • the use of antibiotics is an inevitable cause of antibiotic resistance.
  • chemically synthesized disinfectants such as povidone-iodine, may cause hypersensitivity symptoms such as iodine or itching, burning sensation, and dermatitis due to mucosal damage, and hypothyroidism due to increased iodine levels (Int J Environ Res Public Health. 2019 Oct; 16(20): 3859.).
  • the normal vaginal flora is mostly aerobic and consists of an average of six different bacteria, the most common of which is Lactobacillus, which secretes lactic acid and hydrogen peroxide.
  • the vaginal microflora is determined by various factors that affect the survival of bacteria, including the acidity of the vagina and nutrients necessary for the metabolism of bacteria.
  • vaginal acidity is less than 4.5 and is maintained mainly by lactic acid produced by lactic acid bacteria.
  • Vaginal epithelium is induced by estrogen and is rich in glycogen, which is decomposed into monosaccharides or lactic acid by lactic acid bacteria.
  • the inventors of the present application completed the present invention by isolating a novel strain of the genus Lactobacillus for the prevention or treatment of vaginitis, and confirming the effect of preventing or treating vaginitis thereby.
  • An object of the present invention is to recognize the problems of the prior art mentioned above, and to provide a novel strain of the genus Lactobacillus that can be used for the treatment of vaginitis.
  • An object of the present invention is to provide a culture of a strain of the genus Lactobacillus.
  • An object of the present invention is to provide a composition for preventing or treating vaginitis comprising a Lactobacillus genus strain or a culture thereof.
  • the present invention provides a Lactobacillus crispatus ID-B02 strain of Accession No. KCTC 14466BP.
  • the present invention also provides a Lactobacillus crispatus ID-B05 strain of accession number KCTC 14469BP.
  • the present invention also provides a Lactobacillus crispatus ID-A07 strain of Accession No. KCTC 14470BP.
  • the present invention also provides two or more Lactobacillus crispatus strains selected from the group consisting of:
  • the present invention also provides a culture of two or more Lactobacillus crispatus strains selected from the group consisting of:
  • the present invention also provides a pharmaceutical composition for preventing or treating vaginitis comprising the Lactobacillus crispatus strain or culture thereof.
  • the present invention further provides a health functional food composition for preventing or treating vaginitis comprising the Lactobacillus crispatus strain or culture thereof.
  • Figure 1 shows the results of culturing strains using glucose and glycogen as carbon sources.
  • Vaginitis includes vaginitis and atrophic vaginitis caused by infection with harmful microorganisms, and bacterial, fungal and protozoal vaginitis caused by infection with harmful microorganisms account for most of them. Therefore, it is important to identify and suppress pathogenic microorganisms that cause bacterial, fungal and protozoal vaginitis. Most vaginitis treatments currently on the market are composed of chemicals such as antibiotics, and have excellent short-term effects, but have a high possibility of recurrence.
  • the inventors of the present application isolated beneficial bacteria from vaginal samples of healthy women, and then selected bacteria that can show high viability in the vaginal environment and inhibit vaginitis-causing bacteria, and then selected three novel Lactobacillus crispatus strains. was derived.
  • the main bacterial energy source in the female vagina is glycogen present in the vaginal mucosa, and accordingly, glycogen utilization was confirmed.
  • the ability to generate hydrogen peroxide was confirmed to reveal the antibacterial mechanism related to the ability to compete/inhibit harmful bacteria.
  • the present invention provides the following novel Lactobacillus crispatus strain in one aspect:
  • the present invention provides a culture of the following novel Lactobacillus crispatus strain.
  • the present invention provides two or more Lactobacillus crispatus strains selected from the group consisting of:
  • the present invention provides a culture of two or more Lactobacillus crispatus strains selected from the group consisting of the following.
  • Lactobacillus crispatus Three strains of Lactobacillus crispatus according to the present invention were deposited with the Korea Research Institute of Bioscience and Biotechnology as of February 04, 2021.
  • the Lactobacillus crispatus strain of accession number KCTC 14466BP is described as Lactobacillus crispatus ID-B02
  • the Lactobacillus crispatus strain of accession number KCTC 14469BP is described as Lactobacillus crispatus ID-B05
  • the Lactobacillus crispatus strain of accession number KCTC 14470BP is described as Lactobacillus crispatus ID-A07.
  • the present invention provides a culture of two or more Lactobacillus crispatus strains selected from the group consisting of:
  • culture may mean that each of the three strains or a strain containing one or more of the three strains is cultured in a culture medium or culture medium.
  • the culture may or may not contain each of the three strains or one or more of the three strains.
  • the culture may be a liquid or solid formulation, but is not limited thereto.
  • Various forms of the culture may be included, for example, a concentrate, dried product or extract of the culture.
  • the extract may be extracted using water, an organic solvent, and the like, for example, water, lower alcohol having 1 to 4 carbon atoms, hexane, chloroform, ethyl acetate, or a mixed solvent thereof.
  • the present invention provides a composition for preventing or treating vaginitis comprising the Lactobacillus crispatus strain or culture thereof.
  • a composition for preventing or treating vaginitis comprising the Lactobacillus crispatus strain or culture thereof.
  • Various forms of the culture may be included, for example, a concentrate, dried product or extract of the culture.
  • the strains may be transported in pharmaceutically acceptable carriers such as colloidal suspensions, powders, saline solutions, lipids, liposomes, microspheres, or nanospheres. They may be complexed with or associated with the delivery vehicle and are known in the art such as lipids, liposomes, microparticles, gold, nanoparticles, polymers, condensation reagents, polysaccharides, polyamino acids, dendrimers, saponins, adsorption enhancing substances or fatty acids. It can be delivered in vivo using known delivery systems.
  • pharmaceutically acceptable carriers such as colloidal suspensions, powders, saline solutions, lipids, liposomes, microspheres, or nanospheres. They may be complexed with or associated with the delivery vehicle and are known in the art such as lipids, liposomes, microparticles, gold, nanoparticles, polymers, condensation reagents, polysaccharides, polyamino acids, dendrimers, saponin
  • the composition for preventing or treating vaginitis may be a pharmaceutical composition.
  • the present invention provides a pharmaceutical composition for preventing or treating vaginitis comprising the Lactobacillus crispatus strain or culture thereof.
  • the vaginitis may be induced by at least one microorganism selected from the group consisting of bacterial, fungal and protozoal microorganisms.
  • the pharmaceutical composition may be prepared in unit dosage form by formulation using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily performed by those skilled in the art, or It can be prepared by incorporating into a dose container.
  • the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet, capsule, or gel (eg, hydrogel), and may additionally contain a dispersing agent or a stabilizer. there is.
  • Pharmaceutically acceptable carriers include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia, gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrol, commonly used in formulations. lidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but are not limited thereto.
  • lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, and the like may be further included.
  • the pharmaceutical composition can be administered orally or parenterally and can be used in the form of a general pharmaceutical preparation. That is, the pharmaceutical composition of the present invention can be administered in various oral and parenteral formulations at the time of actual clinical administration.
  • commonly used fillers, extenders, binders, wetting agents, disintegrants It is prepared using excipients.
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations are prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc. do.
  • lubricants such as magnesium styrate and talc are also used.
  • Liquid formulations for oral administration include suspensions, internal solutions, emulsions, syrups, etc.
  • various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included.
  • Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories.
  • Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents.
  • Witepsol Macrogol, Tween 61, cacao butter, laurin paper, glycerol, gelatin, and the like may be used.
  • the concentration of the active ingredient included in the composition may be determined in consideration of the purpose of treatment, the condition of the patient, the required period, and the like, and is not limited to a specific range of concentration.
  • the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount.
  • 'pharmaceutically effective amount means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is based on the type, severity, activity of the drug, drug It may be determined according to factors including sensitivity to , 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 pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent, or in combination with other therapeutic agents, and may be administered simultaneously, separately, or sequentially with conventional therapeutic agents, and may be administered single or multiple times. It is important to administer the amount that can obtain the maximum effect with the minimum amount without side effects in consideration of all the above factors, which can be easily determined by those skilled in the art.
  • the effective amount may vary depending on the patient's age, sex, condition, weight, absorption rate, inactivation rate, excretion rate, disease type, concomitant drug, administration route, severity of vaginitis, gender, weight, age, etc. may increase or decrease accordingly.
  • composition for preventing or treating vaginitis may be a quasi-drug composition.
  • Quasi-drug refers to items that are less active than pharmaceuticals among items used for the purpose of diagnosing, treating, improving, mitigating, treating or preventing human or animal diseases. Products used for the treatment or prevention of diseases in humans and animals, products with minor or no direct action on the human body, etc. may be included.
  • the composition for preventing or treating vaginitis may be a health functional food composition.
  • the present invention provides a health functional food composition for preventing or treating vaginitis comprising the Lactobacillus crispatus strain or culture thereof.
  • the vaginitis may be induced by at least one microorganism selected from the group consisting of bacterial, fungal and protozoal microorganisms.
  • the health functional food composition When the composition is used as a food additive, the health functional food composition may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to conventional methods.
  • the amount of the active ingredient can be appropriately used depending on the purpose of its use (prevention or improvement).
  • health functional food there is no particular limitation on the type of health functional food.
  • foods to which the health functional food composition can be added include meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea There are drinks, alcoholic beverages, vitamin complexes, and the like, and may include all health foods in a conventional sense.
  • the health functional food composition may be prepared as a food, particularly a functional food.
  • the functional food of the present invention includes components commonly added during food preparation, and may include, for example, proteins, carbohydrates, fats, nutrients, and seasonings.
  • natural carbohydrates or flavoring agents may be included as additional ingredients in addition to active ingredients.
  • the natural carbohydrates are monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, etc.), oligosaccharides, polysaccharides (eg, dextrins, cyclodextrins, etc.) or sugar alcohols (eg, maltose, sucrose, etc.) , xylitol, sorbitol, erythritol, etc.) are preferred.
  • natural flavoring agents eg, thaumatin, stevia extract, etc.
  • synthetic flavoring agents eg, saccharin, aspartame, etc.
  • various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonic acid A carbonation agent used in beverages may be further included.
  • composition for preventing or treating vaginitis may be a food composition.
  • composition for preventing or treating vaginitis may be a parenteral composition.
  • composition for preventing or treating vaginitis may be a detergent composition.
  • the composition for preventing or treating vaginitis may be a cosmetic composition.
  • the cosmetic composition may be prepared in general emulsified and solubilized formulations.
  • cosmetic lotion such as softening lotion or nourishing lotion
  • emulsions such as facial lotion and body lotion
  • creams such as nourishing creams, moisture creams, and eye creams; essence; cosmetic ointment; spray; gel; pack; sunscreen; makeup base; foundations such as liquid type, solid type or spray type; powder
  • makeup removers such as cleansing creams, cleansing lotions, and cleansing oils
  • it may be formulated as a cleanser such as a cleansing foam, soap, body wash, etc., but is not limited thereto.
  • the composition for preventing or treating vaginitis may be a composition for external application. It can be used in appropriate combination with common ingredients formulated in the composition for external application for skin, for example, antibiotics, binders, disintegrants, diluents, lubricants, stabilizers, preservatives, or flavoring agents.
  • the skin external preparation may be formulated as an ointment, patch, gel, cream or spray, but is not limited thereto.
  • composition for preventing or treating vaginitis may be a composition for inhibiting the growth of at least one microorganism selected from the group consisting of bacterial, fungal and protozoal microorganisms.
  • the present invention provides a medical device for preventing or treating vaginitis comprising the Lactobacillus crispatus strain or culture thereof.
  • the medical devices are gynecological medical devices for vaginitis, such as colposcopes, gynecological selfchecking colposcopes, vaginal dilators, vaginal speculums, and pelvic inflammatory diseases. It may include a treatment device, a gynecological irrigator, or an antibacterial device for external gynecological use, but is not limited thereto.
  • the present invention provides a disinfectant product for preventing or treating vaginitis containing the Lactobacillus crispatus strain or culture thereof.
  • the above disinfection products are gynecological disinfection products for vaginitis, such as mucosa disinfectant, mucosal disinfection ointment, gynecological disinfection protection pad, gynecological disinfection tissue, and antibacterial gel for gynecological external use. , antibacterial ointment for gynecological external use, or gynecological antiseptic, but is not limited thereto.
  • the present invention provides a feminine cleanser for preventing or treating vaginitis comprising the Lactobacillus crispatus strain or culture thereof.
  • a total of 285 strains were isolated from vaginal fluid samples taken from healthy subjects at Eulji University Hospital. Then, from the vaginal-derived microbial library, bacteria expected to be related to vaginitis prevention and treatment were selected through microbiome analysis. The selection criterion was to ensure novelty and originality as much as possible by considering it as the same strain if it was derived from the same sample regardless of the separation conditions (media, culture method, etc.). The isolated strain was prepared as a glycerol stock and stored at -80 °C.
  • Example 2 Identification of 16S rDNA of the three strains selected in the present invention
  • the analyzed base sequence is identified using the Nucleotide BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi) database of NCBI (National Center for Biotechnology Information) using the homology of the base sequence.
  • ID-B02, ID-B05, and ID-A07 strains showed 99%, 100%, and 99% homology with the Lactobacillus crispatus ATCC 33820 (Accession No. NR_041800) strain, respectively, and all three strains were Lactobacillus crispatus. (Lactobacillus crispatus). Novel Lactobacillus crispatus ID-A07, ID-B02 and ID-B05 strains according to the present invention were deposited with the Korea Center for Biological Resources (Table 1).
  • test strains were inoculated into MRS (de Man, Rogosa and Sharpe) medium, incubated at 37°C for 24 hours, and then plated on Brucella agar medium containing TMB (tetramethyl-benzidine) and horseradish peroxidase at 37°C. were cultured anaerobically for 2 days.
  • MRS de Man, Rogosa and Sharpe
  • TMB tetramethyl-benzidine
  • test strains were inoculated into NYC III (New York City III) medium, incubated at 37 ° C for 24 hours, washed twice with PBS (Phosphate-buffered saline), suspended in NYC III medium, and 0.5% glycogen was used as a carbon source. Inoculated in the included NYC III medium and cultured for 48 hours, absorbance was measured at 600 nm and shown in Table 3.
  • NYC III New York City III
  • PBS Phosphate-buffered saline
  • Example 5 Inhibiting the growth of vaginitis-related harmful microorganisms
  • GV Gardnerella vaginalis
  • CA Candida albicans
  • PBS phosphate-buffered saline
  • Lactobacillus crispatus strains AB70 (accession number: KCTC12976BP) and SNUV220 (accession number: KCTC18374P), selected in Example 2 for glycogen utilization and vaginal harmful microorganisms Inhibitory efficacy was compared.
  • As representative harmful microorganisms related to vaginitis Gardnerella vaginalis (GV), Candida albicans (CA), and Trichomonas vaginalis (TV) were used.
  • GV Gardnerella vaginalis
  • CA Candida albicans
  • TV Trichomonas vaginalis
  • the test strains showed relatively poor glycogen utilization and ability to inhibit harmful microorganisms in the previous experiment.
  • Lactobacillus crispatus V22 strain which was not selected by the above method, was used.
  • each strain was inoculated into a medium supplemented with 0.5% glucose and 0.5% and 1% glycogen as carbon sources, and cultured at 37°C for 18 hours. .
  • the absorbance of each culture end solution was measured, the absorbance of the culture solution without a carbon source was subtracted, and then the proliferation rate by the corresponding carbon source for each strain was compared.
  • the selected strains showed a higher growth rate in a medium containing 1% glycogen than in a medium containing 0.5% glucose, whereas SNUV220 did not. Therefore, it can be seen that the strains selected in the present invention are superior to the previously patented strains in terms of their ability to use glycogen as a carbon source. This excellent glycogen availability is advantageous for the formation of colonies by proliferating through the glycogen present in the vagina, so it seems to have a high vaginal survival rate.
  • Example 8-2 Inhibition of vaginitis-related harmful microorganisms
  • each strain was inoculated and cultured in MRS medium containing 1% glycogen as a carbon source, and the supernatant was collected by centrifugation. Harmful microorganisms were cultured in a medium containing 50% of the culture supernatant of the strain to confirm the growth inhibitory ability.
  • the types of harmful microorganisms used in the test, culture conditions, and growth measurement methods are shown in Table 6 below.
  • V22 which is a dropout strain that was not selected in the present invention, it showed the lowest inhibition rate against all harmful microorganisms.
  • SNUV220 showed a higher inhibition rate than AB70 in the culture of all harmful microorganisms, and all three selected strains showed higher inhibition rates than these two patented strains.
  • TV the growth was completely inhibited when the supernatant of the selected strain was treated, which seems to have a high overall inhibition rate because TV has sensitive culture conditions. However, it seems sufficient to compare the growth inhibition ability between strains.
  • the selected strains of the present invention have better glycogen utilization ability and vaginal harmful microorganism inhibition ability than the previously invented Lactobacillus crispatus strains under conditions where glycogen can be used as a carbon source, such as in the vagina.
  • three novel Lactobacillus crispatus strains are classified based on glycogen availability, which is a major bacterial energy source in the vagina, antibacterial activity against harmful microorganisms, and whether the growth of harmful bacteria can be inhibited in the vaginal environment. was isolated, and the isolated strain can inhibit vaginitis-causing bacteria to exhibit the antibacterial effect required for the prevention or treatment of vaginitis.

Abstract

La présente invention concerne une nouvelle souche de Lactobacillus sp., une culture de celle-ci et une composition pour traiter la vaginite la comprenant et, en particulier, une nouvelle souche Lactobacillus crispatus isolée à partir d'un échantillon vaginal d'une femme en bonne santé, une culture de celle-ci et une composition pharmaceutique et une composition d'aliments fonctionnels pour la santé pour la prévention ou le traitement de la vaginite la comprenant. Selon la présente invention, trois types de nouvelles souches de Lactobacillus crispatus ont été isolées sur la base de leur capacité à utiliser le glycogène (source d'énergie bactérienne majeure dans le vagin), de leur activité antibactérienne contre les micro-organismes nuisibles et de leur capacité à inhiber les bactéries nuisibles dans l'environnement vaginal. Les souches isolées suppriment les bactéries responsables de la vaginite, présentant ainsi l'effet antibactérien souhaité nécessaire à la prévention ou au traitement de la vaginite.
PCT/KR2022/016340 2021-10-26 2022-10-25 Souche de lactobacillus crispatus et composition pour la prévention ou le traitement de vaginite la comprenant WO2023075357A1 (fr)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0872231A1 (fr) * 1997-04-14 1998-10-21 Dr. A. Tosi Farmaceutici S.R.L. Compositions pharmaceutiques à base de lactobacilles pour l'administration transmucosale
US6372209B1 (en) * 1997-04-11 2002-04-16 Gyne Logix, Inc. Vaginal lactobacillus medicant
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EP0872231A1 (fr) * 1997-04-14 1998-10-21 Dr. A. Tosi Farmaceutici S.R.L. Compositions pharmaceutiques à base de lactobacilles pour l'administration transmucosale
KR20100079078A (ko) * 2008-12-30 2010-07-08 전남대학교산학협력단 락토바실러스 퍼멘툼 Ｎｏ．１９６９ 균주를 유효성분으로 포함하는 세균성질염치료용 질내 투여용 약학조성물
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