Feed, antifungal or pharmaceutical composition comprising the strain of Lactobacillus crispatus KBL693

Abstract

"Lactobacillus crispatus kbl693 strain, food composition, feed or antifungal, external skin preparation, medical plaster, pharmaceutical composition and cosmetic composition comprising said strain and therapeutic use of said composition." The present invention relates to a Lactobacillus crispatus kbl693 strain and its use. The Lactobacillus crispatus kbl693 strain (accession no. kctc 13519bp) according to the present invention attenuates allergic cell reactions, significantly improves the symptoms of atopic dermatitis and exhibits anti-inflammatory and antifungal effects. Thus, the single strain alone can achieve all the objectives of alleviating atopic dermatitis and other allergic diseases and improving inflammatory and autoimmune diseases, thus finding advantageous applications as a probiotic substance.Furthermore, based on its antifungal activity, the strain can be advantageously used in an external skin preparation against various skin diseases caused by fungi, and in a cosmetic composition and a functional plaster for the relief of sensitive skin.

Description

1 / 42 “FEED, ANTIFUNGAL OR PHARMACEUTICAL COMPOSITION COMPRISING THE LACTOBACILLUS CRISPATUS KBL693 STRAIN” FIELD OF THE INVENTION

[001] The present invention relates to a strain of Lactobacillus crispatus KBL693 and its use. More specifically, the present invention relates to a healthy functional food composition having at least one effect selected from the group consisting of relief of allergic symptoms, relief of inflammatory symptoms, and immunoregulation; an antifungal composition; and a pharmaceutical composition for the treatment of at least one disease selected from the group consisting of allergic diseases including atopic dermatitis and / or inflammatory or autoimmune diseases, comprising at least one selected from the group consisting of a novel probiotic strain of Lactobacillus crispatus KBL693, cultures of said strain, lysates of said strain, and extracts of said strain. FUNDAMENTALS OF THE INVENTION

[002] Probiotics refer to microorganisms and their resulting products that have antimicrobial and enzymatic activities to help balance intestinal microorganisms. Furthermore, probiotics are also defined as live bacteria in the form of one or more strains to improve intestinal flora when provided to humans or animals in the form of dried cells or fermentation products. Probiotics must inhabit the human intestine, be non-pathogenic and non-toxic, and survive long enough to reach the intestine. Additionally, probiotics must maintain viability and activity until consumed in delivered food, be sensitive to antibiotics used to prevent infection, and not have antibiotic-resistant plasmids. Moreover, probiotics must be resistant to acids, enzymes, and bile in the intestinal environment.

[003] These probiotics may include, for example, Bacillus sp., which has an excellent capacity to produce digestive enzymes, such as amylase, protease, Petition 870260058402, dated 06 / 16 / 2026, page 11 / 106 2 / 42 lipase, cellulase, and phosphatase, Lactobacillus sp. producing lactic acid, and photosynthetic bacteria preventing odor by using odor-causing substances (such as ammonia, hydrogen sulfide, and amines) that remain in animal feces during the metabolic process. Recently, probiotics have been reported to have several health-enhancing effects, including improved gut health, and therefore stand out as key therapeutic substances that can replace existing compound-based therapeutic agents.

[004] Meanwhile, allergy is a biochemical phenomenon that exhibits a unique and altered response to a foreign substance (antigen, allergen). The foreign substance that causes the symptoms is called an allergen, while the diseases resulting from these symptoms are called allergic diseases. Allergy is a pathological process in the living body resulting from the antigen-antibody reaction. In general, there are four types of allergy depending on the triggering period of the reaction and the involvement of complement. Type 1, among these, is the anaphylactic type (immediate type) in which the target organs are mainly digestive organs, skin and lungs, and common symptoms include gastrointestinal allergy, urticaria, atrophodermatitis, allergic rhinitis and bronchial asthma, etc.The pathological mechanism of Type 1 is known as follows: when antigens come into contact with IgE antibodies attached to the surface of mast cells and basophilic leukocytes, the target cells are activated to secrete chemical transmitters, such as histamine, leukotriene, and PAF, and then blood vessels and smooth muscles are constricted. This mechanism can often be combined with Type 4 (delayed type). In other words, this anaphylaxis and allergic reaction can arise due to a variety of alterations in mast cells, etc. Mast cell activation, which leads to degranulation, is caused by the binding of antigen, anti-IgE, lectin, etc. to Fc receptors, stimulation by anaphylatoxin, etc., or other drugs such as calcium ionophore, compound 48 / 80, codeine, and hormones. Petition 870260058402, dated 06 / 16 / 2026, page 12 / 106 3 / 42 synthetic adrenocorticotropic.

[005] Mast cells and basophilic leukocytes in the blood are known as the body's main cells for causing many allergic diseases, such as allergic rhinitis, allergic dermatitis, asthma, food allergy, and anaphylactic shock. These cells have receptors (FcRI) on their surfaces for IgE, which is an allergy-causing antibody, and the cells are stimulated by allergy-causing substances (antigen, allergen) to secrete their own various allergy-causing substances out of the cells (Kim K et al., Eur J Pharmacol, 581: 191-203, 2008).

[006] Among allergic diseases, atopic dermatitis, widely known to the public, is a chronic recurrent skin disease that affects newborns or children and can persist into adulthood. Like asthma or allergic rhinitis, atopic dermatitis is an inflammatory skin disease associated with local infiltration of T lymphocytes that produce IL-4 and IL-5. IL-4, well known to the public, controls the development of the T helper 2 (Th2) phenotype, resulting in overproduction of immunoglobulins (Ig) and eosinophilia, and increased serum IgE levels. 80 to 90% of individuals with positive skin tests for food and inhalant allergens presented with atopic dermatitis.

[007] There are different treatments to treat or prevent allergic diseases and atopic dermatitis, but no effective treatment has yet been found. Some drug-based treatments are known, but even short-term administration of the drug for treatment would develop a tolerance, and long-term administration can cause serious side effects, and therefore such drug-based treatments for allergic diseases and atopic dermatitis have recently been avoided. Under these circumstances, without the treatment having any obvious and absolute effect, the irritating symptoms, such as itching and redness of the skin, in addition to allergy, often do not improve. Petition 870260058402, dated 06 / 16 / 2026, page 13 / 106 4 / 42

[008] Under these circumstances, the present inventors dedicated themselves to studies of probiotics to find a way to replace drug-based treatments for allergic diseases, including atopic dermatitis, which do not have satisfactory treatments. Then the present invention was completed by confirming that a new strain of Lactobacillus crispatus showed excellent therapeutic effects in allergic diseases, such as atopic dermatitis, and also confirming that said strain also showed superior effects in anti-inflammatory, immunoregulatory and antifungal activities. SUMMARY OF THE INVENTION

[009] The objective of the present invention is to provide a new strain showing excellent effects in relieving allergic symptoms, such as atopic dermatitis, relieving inflammatory and immunoregulatory symptoms, and a variety of uses thereof.

[010] In order to achieve the objective, the present invention provides the Lactobacillus crispatus KBL693 strain (KCTC Accession No. 13519BP).

[011] In addition, the present invention provides a food composition or food additive composition comprising at least one selected from the group consisting of said strain, cultures of said strain, lysates of said strain and extracts of said strain.

[012] The present invention also provides a feed composition or feed additive composition comprising at least one selected from the group consisting of said strain, cultures of said strain, lysates of said strain and extracts of said strain.

[013] The present invention also provides an antifungal composition, such as an anti-dandruff composition, comprising at least one selected from the group consisting of said strain, cultures of said strain, lysates of said strain and extracts of said strain. Petition 870260058402, dated 06 / 16 / 2026, page 14 / 106 5 / 42

[014] The present invention also provides a pharmaceutical composition for the treatment of allergic diseases, such as atopic dermatitis, and / or inflammatory or autoimmune diseases, comprising at least one selected from the group consisting of said strain, cultures of said strain, lysates of said strain and extracts of said strain.

[015] The present invention also provides a method for treating allergic diseases, including atopic dermatitis, and / or inflammatory or autoimmune diseases, comprising administering at least one selected from the group consisting of said strain, cultures of said strain, lysates of said strain, and extracts of said strain to an individual in need of treatment.

[016] The present invention also provides a composition comprising at least one selected from the group consisting of said strain, cultures of said strain, lysates of said strain and extracts of said strain, for use in the prevention or treatment of allergic diseases, including atopic dermatitis, and / or inflammatory or autoimmune diseases.

[017] The present invention also provides the use of a composition for preparing a preventive or therapeutic drug for allergic diseases, including atopic dermatitis, and / or inflammatory or autoimmune diseases, comprising at least one selected from the group consisting of said strain, cultures of said strain, lysates of said strain and extracts of said strain.

[018] The present invention also provides a cosmetic composition comprising at least one selected from the group consisting of said strain, cultures of said strain, lysates of said strain and extracts of said strain.

[019] The present invention also provides a cosmetic plaster or a medical plaster comprising at least one selected from the group consisting of said strain, cultures of said strain, lysates of said strain and extracts of said strain. Petition 870260058402, dated 06 / 16 / 2026, p. 15 / 106 6 / 42 BRIEF DESCRIPTION OF THE DRAWINGS

[020] Figure 1 illustrates the result of comparing the inhibitory effect on histamine secretion by treating various Lactobacillus strains, including the Lactobacillus crispatus KBL693 strain, and the antihistamine ketotifen, after inducing histamine production by antigen-antibody reaction in RBL 2H3 cell lines.

[021] Figure 2 illustrates the result that confirms the inhibitory effect of IL-4 expression by several Lactobacillus strains, including the Lactobacillus crispatus KBL693 strain, after inducing an allergic reaction in EL4 cell lines.

[022] Figure 3 illustrates the result that confirms the inhibitory effect of IL-5 expression by several Lactobacillus strains, including the Lactobacillus crispatus KBL693 strain, after inducing an allergic reaction in EL4 cell lines.

[023] Figure 4 illustrates the result that confirms an inhibitory effect on IgE secretion by several Lactobacillus strains, including the Lactobacillus crispatus KBL693 strain, after induction of an allergic reaction in U266B1 cell lines.

[024] Figure 5 illustrates the result of observing the remarkable effect of increasing the amount of anti-inflammatory cytokine IL-10 secretion by treating the KBL693 strain, when treated with various Lactobacillus strains after inducing an inflammatory reaction in RAW 264.7 cell lines.

[025] Figure 6 illustrates the result that confirms the remarkable immunoregulatory and anti-inflammatory effect of treating the KBL693 strain with an IL10 / TNF-α value when treated with various Lactobacillus strains after inducing an inflammatory reaction in RAW 264.7 cell lines.

[026] Figure 7 illustrates the result that confirms the remarkable immunoregulatory and anti-inflammatory effect of treating the KBL693 strain with an IL-10 / IL-6 value, when treated with various Lactobacillus strains after inducing an inflammatory reaction in RAW 264.7 cell lines. Petition 870260058402, dated 06 / 16 / 2026, page 16 / 106 7 / 42

[027] Figure 8 illustrates the result of a spot test that confirms the antifungal activity of the Lactobacillus crispatus KBL693 strain.

[028] Figure 9 illustrates the result that confirms the effect of reducing dermatitis scores by oral administration of the Lactobacillus crispatus KBL693 strain to mouse models in which atopic dermatitis was induced.

[029] Figure 10 illustrates the result that confirms the itch-relieving effect of oral administration of the Lactobacillus crispatus KBL693 strain to mouse models in which atopic dermatitis was induced.

[030] Figure 11 illustrates the result that confirms the effect of reducing ear thickness by oral administration of the Lactobacillus crispatus KBL693 strain to mouse models in which atopic dermatitis was induced.

[031] Figure 12 illustrates the result that confirms the skin thickness reduction effect of oral administration of the Lactobacillus crispatus KBL693 strain to mouse models in which atopic dermatitis was induced.

[032] Figure 13 (A) depicts the experimental design using ovalbumin (OVA)-induced asthma mouse models, and Figure 13 (B) illustrates the observation of lung tissues collected from each group of the normal control group (Control-PBS), control group with induced asthma (OVA-PBS), and test group (OVA-LC206) by an optical microscope after H&E staining.

[033] Figure 14 illustrates the result of determining the expression level of total IL-5 and total IL-13 in lung tissues collected from each normal control group (Control-PBS), control group with induced asthma (OVA-PBS) and test group (OVA-KBL693).

[034] Figure 15 illustrates the result that confirms the effect of reducing airway hyperresponsiveness (AHR) by the Lactobacillus crispatus KBL693 strain administered to mouse models with house dust mite-induced asthma (HDM). Petition 870260058402, dated 06 / 16 / 2026, page 17 / 106 8 / 42

[035] Figure 16 illustrates the result that confirms the effect of the reduction of eosinophils, CD4 IL-5+ T cells and CD4 IL-13+ T cells by the administration of the Lactobacillus crispatus KBL693 strain. DETAILED DESCRIPTION

[036] Unless otherwise defined, all technical and scientific terms used in this specification mean the same as understood by a person skilled in the art (“those skilled in the art”). In general, the nomenclature used in this specification is well known in the art and commonly used.

[037] The present invention has discovered an anti-allergic effect of microorganisms derived from the human body, and has selected the Lactobacillus crispatus KBL693 strain (Accession No. KCTC 13519BP) having excellent allergy-inhibiting effects. Analysis of the 16S rDNA of said strain demonstrates that said strain is a novel strain that has never been known to the public.

[038] According to one embodiment of the present invention, the present invention relates to a novel probiotic strain of Lactobacillus crispatus KBL693 (KCTC Accession No. 13519BP), and said strain is characterized by comprising the 16S rDNA sequence of SEQ ID NO: 1. < SEQ ID NO: 1 > 16S rDNA sequence of the Lactobacillus crispatus strain KBL693 (KCTC Accession No. 13519BP) CGCGGGGTGGCGCGAGCTATAATGCAGTCGAGCGAGCGGAACTAACA GATTTACTTCGGTAATGACGTTAGGAAAGCGAGCGGCGGATGGGTGAGTAACA CGTGGGGAACCTGCCCCATAGTCTGGGATACCACTTGGAAACAGGTGCTAATA CCGGATAAGAAAGCAGATCGCATGATCAGCTTTTAAAAGGCGGCGTAAGCTGTC GCTATGGGATGGCCCCGCGGTGCATTAGCTAGTTGGTAAGGTAAAGGCTTACC AAGGCGATGATGCATAGCCGAGTTGAGAGACTGATCGGCCACATTGGGACTGA GACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGA Petition 870260058402, dated 06 / 16 / 2026, p. 18 / 106 9 / 42 CGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAA AGCTCTGTTGTTGGTGAAGAAGGATAGAGGTAGTAACTGGCCTTTATTTGACGG TAATCAACCAGAAAGTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTA GGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGAAGA ATAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGGAACTGCATCGGAAACTGT TTTTCTTGAGTGCAGAAGAGGAGAGTGGAACTCCATGTGTAGCGGTGGAATGC GTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTCTCTGGTCTGCAACTG ACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTC CATGCCGTAAACGATGAGTGCTAAGTGTTGGGAGGTTTCCGCCTCTCAGTGCTG CAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACT CAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGA AGCAACGCGAAGAACCTTACCAGGTCTTGACATCTAGTGCCATTTGTAGAGATA CAAAGTTCCCTTCGGGGACGCTAAGACAGGTGGTGCATGGCTGTCGTCAGCTC GTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTTATTAGT TGCCAGCATTAAGTTGGGCACTCTAATGAGACTGCCGGTGACAAACCGGAGGA AGGTGGGGATGACGTCAAGTCATCATGCCCCTTATGACCTGGGCTACACACGT GCTACAATGGGCAGTACAACGAGAAGCGAGCCTGCGAAGGCAAGCGAATCTCT GAAAGCTGTTCTCAGTTCGGACTGCAGTCTGCAACTCGACTGCACGAAGCTGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTG TACACACCGCCCGTCACACACCATGGGAGTCTGCAATGCCCAAAGCCGGTGGCCT AACCTTCGGAAGGAGCCGTCTAAGTAGACAATGTGCA

[039] Subsequently, the present invention conducted experiments regarding the efficacy of said strain and thus verified that this strain has an excellent inhibitory effect on allergies, such as atopic dermatitis, has immunoregulatory properties, relieves the inflammatory reaction, and has antifungal activity. Furthermore, the inventors confirmed that said effects can be provided not only in the condition of live bacteria, but also under low-temperature sterilization or Petition 870260058402, dated 06 / 16 / 2026, p. 19 / 106 10 / 42 high-temperature sterilization.

[040] Consequently, in another embodiment of the present invention, the present invention relates to a food composition or food additive composition comprising at least one selected from the group consisting of the Lactobacillus crispatus KBL693 strain (KCTC Accession No. 13519BP), a cell component of said strain, cultures of said strain, lysates of said strain and extracts of said strain.

[041] The composition can be characterized as being a healthy functional food composition having at least one effect selected from the group consisting of relief of allergic symptoms, such as atopic dermatitis, relief of inflammatory symptoms, and immunoregulation.

[042] The said food composition or food additive composition can be easily used as an effective food for the relief of allergic symptoms, such as atopic dermatitis, relief of inflammatory symptoms, and / or immunoregulation, and for the prevention thereof, for example, as main ingredients or minor ingredients of foods, food additives, composition of healthy functional foods or functional drinks, but not limited to them.

[043] The term “food composition” refers to a natural or artificial product comprising at least one nutrient and, more preferably, refers to a product that has been made edible through certain processing, generally encompassing all foods, food additives, functional health foods and functional beverages.

[044] The food that may comprise said food composition according to the present invention as an additive may include, for example, different types of foods, beverages, chewing gum, tea, vitamin complex or functional food. In addition, the food of the present invention includes special nutritional foods (e.g., modified milk, foods for newborns / babies), Petition 870260058402, dated 06 / 16 / 2026, page 20 / 106 11 / 42 Processed meat products, fish meat products, tofu, muk, noodles (e.g., ramen, Asian noodles), bakery products, health food supplements, seasoning products (e.g., soy sauce, soy paste, red pepper paste, mixed paste), sauces, confectionery (e.g., snacks), sweets, chocolates, chewing gum, ice cream, dairy products (e.g., fermented milk, cheese), other processed foods, kimchi, savory foods (e.g., different types of kimchi, pickled foods), beverages (e.g., fruit juice, vegetable juice, soy milk, fermented beverages), and natural seasonings (e.g., ramen broth powder), but not limited to these. The said foods, beverages, or food additives may be prepared in conventional ways.

[045] The term “functional health food” is a group of foods to which value is added so that its function is exercised and expressed for a predetermined purpose, using physical, biochemical or bioengineering techniques, or a processed food designed so that the in vivo adjustment functions of the relevant food composition, such as rhythm adjustment in prophylaxis, disease prevention and disease recovery, are sufficiently expressed. Such functional food may comprise food supplement additives that are scientifically acceptable in food terms and may additionally comprise suitable carriers, excipients and diluents, which are normally used in its manufacture.

[046] The term “functional beverages”, as used in the present invention, refers collectively to beverage products for relieving thirst or for enhancing taste. There is no particular limitation to this, except that, as essential ingredients in the indicated proportion, a composition for the relief of allergic symptoms, such as atopic dermatitis, relief of inflammatory symptoms, and / or immunoregulation and prevention thereof must be included in the beverages, and various flavoring agents. Petition 870260058402, dated 06 / 16 / 2026, p. 21 / 106 12 / 42 or natural carbohydrates may be contained in them as additional ingredients, just like in regular drinks.

[047] In addition to the above, the food comprising the food composition or food additive composition according to the present invention may contain various nutrients, vitamins, minerals (electrolyte), flavoring agents, such as synthetic flavoring agents and natural flavoring agents, coloring agents and fillers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickening agents, pH control agents, stabilizing agents, preservatives, glycerin, alcohol, carbonating agents as used in carbonated beverages and the like, and each of the above ingredients may be used alone or in combination with each other.

[048] In food comprising the food composition according to the present invention, the composition of the present invention may comprise an amount of 0.001% by weight to 100% by weight and, preferably, 1% by weight to 99% by weight, based on the total weight of the food; in the case of beverages, it may comprise an amount of 0.001 g to 10 g, and preferably 0.01 g to 1 g, in relation to 100 ml. For long-term intake for health and hygiene purposes or for health monitoring purposes, however, the amount may be below the range mentioned above; and since the effective ingredients do not present problems in terms of safety profile, they may be used in an amount above the range and are not limited to the quantity range mentioned above.

[049] The food composition according to the present invention may comprise the Lactobacillus crispatus KBL693 strain alone or in combination with an acceptable carrier, or may be prepared in the form of a composition suitable for human or animal consumption. That is, the Petition 870260058402, dated 06 / 16 / 2026, p. 22 / 106 13 / 42 composition may be added to food that does not comprise probiotic bacteria or some probiotic bacteria. For example, the microorganisms that may be used in combination with the strain according to the present invention in the preparation of the food of the present invention must be suitable for consumption by humans or animals, and must have probiotic activities to inhibit harmful pathogenic bacteria or to improve the balance of microorganisms in the intestinal tract of mammals after ingestion, but not limited to that. These probiotic microorganisms may include, for example, yeast, such as Saccharomyces, Candida, Pichia or Torulopsis, fungi such as Aspergillus, Rhizopus, Mucor, or Penicillium, and bacteria belonging to the genera Lactobacillus, Bifidobacterium, Leuconostoc, Lactococcus, Bacillus, Streptococcus, Propionibacterium, Enterococcus, or Pediococcus. Specifically suitable probiotic microorganisms may include, for example, Saccharomyces cerevisiae, Bacillus coagulans, Bacillus licheniformis, Bacillus subtilis, Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium longum, Enterococcus faecium, Enterococcus faecalis, Lactobacillus acidophilus, Lactobacillus alimentarius, Lactobacillus casei, Lactobacillus curvatus, Lactobacillus delbruckii, Lactobacillus johnsonii, Lactobacillus farciminus, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus rhamnosus, Lactobacillus reuteri, Lactobacillus sakei, Lactococcus lactis, or Pediococcus acidilactici. Preferably, the food composition according to the present invention may further comprise a mixture of probiotic microorganisms with excellent probiotic activities and superior anti-allergic, anti-inflammatory and / or immunoregulatory activities to further enhance its effects.The carriers that can be included in the food composition of the present invention may include, for example, extenders, high-fiber additives, encapsulating agents, and lipids, which are widely known in the art. The Lactobacillus crispatus KBL693 strain in the present invention may be... Petition 870260058402, dated 06 / 16 / 2026, p. 23 / 106 14 / 42 in lyophilized or encapsulated form, or in the form of culture suspensions or dry powders.

[050] The composition of the present invention can also be supplied in the form of a food additive comprising said strain or a feed comprising the same.

[051] The feed additive of the present invention may be in the form of a dry or liquid formulation, and may also comprise other non-pathogenic microorganisms in addition to said strain of Lactobacillus crispatus KBL693. Microorganisms that can be added to feed additives may include, for example, Bacillus subtilis, which can produce protease, lipase, and sugar conversion enzymes; a strain of Lactobacillus, which has physiological activity and degradability of organic compounds under anaerobic conditions, such as in the stomach of cows; filamentous fungi such as Aspergillus oryzae, which show effects on increasing animal weight, milk production, and feed digestibility (Slyter, LLJ Animal Sci. 1976, 43, 910-926); and yeasts such as Saccharomyces cerevisiae (Johnson, D. E. et al., J. Anim. Sci., 1983, 56, 735-739; Williams, PEV et al., 1990, 211).

[052] The feed additive of the present invention may further comprise at least one enzymatic agent in addition to said Lactobacillus crispatus KBL693 strain. The additional enzymatic agents may be in dry or liquid form, and may include, for example, steatolytic enzymes, such as lipase, phytase to produce phosphate and inositol phosphate by degradation of phytic acid, amylase, i.e., an enzyme to hydrolyze the α-1,4-glycoside linkage included, for example, in starch and glycogen, phosphatase, i.e., an enzyme to hydrolyze organic phosphoric acid ester, carboxymethyl cellulase to degrade cellulose, xylase to degrade xylose, maltase to hydrolyze maltose into two glucose molecules, and sugar-producing enzymes such as invertase to produce a glucose-fructose mixture by hydrolysis of sucrose. Petition 870260058402, dated 06 / 16 / 2026, page 24 / 106 15 / 42

[053] In using the Lactobacillus crispatus KBL693 strain of the present invention as feed additives, raw feed ingredients, such as peanuts, peas, beets, pulp, grain by-products, powdered animal viscera and powdered fishmeal, including various grains and soy protein, may be used. They may be processed or unprocessed, and may be used without limitation. Processing may include, but is not limited to, a process in which the raw feed ingredients are loaded and compressed under pressure against a given outlet and, for proteins, extrusion by which the proteins are degenerated to increase availability may preferably be used. Extrusion denatures the proteins by means of heat treatment to destroy antienzymatic factors, which is advantageous.Furthermore, for soy proteins, their digestibility can be improved through extrusion to inactivate antinutrients, such as a trypsin inhibitor, one of the protease inhibitors present in soy. Additionally, extrusion can promote improved protease digestibility, increasing the nutritional value of soy proteins.

[054] According to another embodiment of the present invention, the present invention relates to an antifungal composition comprising at least one selected from the group consisting of said strain, cultures of said strain, lysates of said strain and extracts of said strain.

[055] The composition can be characterized by showing antifungal activity against, but not limited to, Malassezia furfur, Malassezia globosa and Malassezia restricta selected from the group consisting of, but not limited to, the same.

[056] The composition may be a composition to prevent, relieve or treat seborrheic dermatitis or dandruff, and said seborrheic dermatitis may be on the scalp.

[057] In addition, the said composition may be a composition for preventing, relieving or treating urticaria, skin rash, tinea corporis, tinea cruris or tinea pedis, Petition 870260058402, dated 06 / 16 / 2026, page 25 / 106 16 / 42 due to fungal infection.

[058] The said strain, and cultures of the said strain, lysates of the said strain and extracts of the said strain may comprise an amount of 0.1% by weight to 50% by weight, based on the total weight of the composition.

[059] The said antifungal composition may be a pharmaceutical composition, a cosmetic composition or a food composition, and may also be an external skin preparation.

[060] The said cosmetic composition may be supplied in all dosage forms suitable for topical application, for example, in the form of a liquid, oil-in-water emulsion, water-in-oil emulsion, suspension, solid, gel, powder, paste, foam or aerosol. The said compositions of the above dosage forms may be prepared by conventional methods used in the art.

[061] In addition to the above ingredients, said composition may comprise other ingredients in an amount that does not impair the main effect, preferably in an amount that provides a synergistic effect on the main effect. The composition according to the present invention may comprise a substance selected from the group consisting of vitamins, polypeptides, polysaccharides and sphingolipids. Furthermore, the cosmetic composition of the present invention may comprise a moisturizer, an emollient, a surfactant, a UV absorber, a preservative, a sterilizer, an antioxidant, a pH adjusting agent, organic and inorganic pigments, a flavoring agent, a cooling agent or an antiperspirant.The percentage of the combination of said ingredients may be selected by those skilled in the art within a range so as not to impair the purpose and effect of the present invention, and may be in a range of 0.01% by weight to 5% by weight and, specifically, from 0.01% by weight to 3% by weight, based on the total weight of the composition.

[062] According to the above modality, the antifungal composition of Petition 870260058402, dated 06 / 16 / 2026, page 26 / 106 17 / 42 The present invention may be an external preparation for the skin, such as a cream, ointment, shampoo or treatment.

[063] According to another embodiment of the present invention, the present invention relates to a pharmaceutical composition for the treatment or prevention of allergic diseases, including atopic dermatitis, and / or inflammatory or autoimmune diseases, comprising at least one selected from the group consisting of a cellular component of the Lactobacillus crispatus KBL693 strain, cultures of said strain, lysates of said strain and extracts of said strain.

[064] The pharmaceutical composition of the present invention may be provided in the form of a live bacterial cell component, dried strain, cultures of said strain, lysates of said strain, or a composition in combination with a pharmaceutically acceptable carrier or medium. The carriers or media that may be used herein may include a solvent, a dispersant, a coating, an enhancer, a controlled-release formulation (i.e., sustained-release formulation), or at least one inert excipient including starch, polyol, granules, microfine cellulose, microcrystalline cellulose, such as Celphere, Celphere microspheres, diluent, lubricant, binder, disintegrant. The tablet of the above composition may, if desired, be coated by a standard aqueous or non-aqueous technique.Examples of pharmaceutically acceptable carriers and excipients for use as pharmaceutically acceptable inert carriers, and said additional ingredients may include, for example, a binder, a filler, a disintegrant, a lubricant, an antimicrobial agent and a coating agent.

[065] In addition, the pharmaceutical composition of the present invention can be used as an external preparation comprising a dosage form selected from the group consisting of ointments, creams, pastes, liquids and solutions for cutaneous application, glycerogelatins, liniments, powders for cutaneous application, aerosols and plasters. Petition 870260058402, dated 06 / 16 / 2026, page 27 / 106 18 / 42

[066] In the present invention, said allergic diseases refer to conditions associated with the expression of IL-4 or IL-5 and may include, for example, eczema, allergic asthma, atopic dermatitis, allergic rhinitis, allergic conjunctivitis, urticaria or anaphylaxis. Preferably, the present invention may be characterized by the fact that the diseases are selected from the group consisting of infantile eczema, allergic asthma, allergic rhinitis, atopic dermatitis, allergic conjunctivitis and food allergy, but not limited to these.

[067] In the present invention, said inflammatory or autoimmune diseases, or symptoms that can be relieved by immunoregulation, are those that can be treated, relieved or prevented by promoting the expression or secretion of IL10, which is an anti-inflammatory immunoregulatory cytokine, and may include, for example, rheumatoid arthritis, rheumatic fever, lupus, systemic scleroderma, atopic dermatitis, psoriasis, psoriatic arthritis, asthma, Guillain-Barré syndrome, myasthenia gravis, dermatomyositis, polymyositis, multiple sclerosis, autoimmune encephalomyelitis, polyarteritis nodosa, Hashimoto's thyroiditis, temporal arteritis, juvenile diabetes, alopecia areata, pemphigus, aphthous stomatitis, autoimmune hemolytic anemia, Wegener's granulomatosis, Sjögren's syndrome, Addison's disease, Crohn's disease, Behcet's disease, edema, conjunctivitis, periodontitis, rhinitis, otitis media, chronic sinusitis, pharyngolaryngitis, tonsillitis, bronchitis, pneumonia, gastric ulcer, gastritis, colitis, gout, eczema,acne, contact dermatitis, seborrheic dermatitis, ankylosing spondylitis, fibromyalgia, osteoarthritis, periarthritis of the shoulder, tendinitis, tenosynovitis, myositis, hepatitis, cystitis, nephritis, sepsis, vasculitis, bursitis and similar conditions, but not limited to these.

[068] The term 'treat', unless otherwise indicated, refers to reversing or alleviating the diseases or conditions used with said term or one or more symptoms thereof, inhibiting their progression or preventing them. The term 'treatment', as used in the present invention, refers to an act of Petition 870260058402, dated 06 / 16 / 2026, p. 28 / 106 19 / 42 'treat' as defined above. Consequently, the treatment or therapeutic regimen of a disease in mammals may include one or more of the following: (1) Inhibit the growth of the disease, that is, inhibit its development (2) Prevent the spread of the disease (3) Relieve the disease (4) Prevent the recurrence of the disease and (5) Palliate the symptoms of the disease

[069] A composition of the present invention for preventing or treating allergic diseases, such as atopic dermatitis, and / or inflammatory or autoimmune diseases, may comprise a pharmaceutically effective amount of the Lactobacillus crispatus KBL693 strain alone or in combination with at least one of pharmaceutically acceptable carriers, excipients or diluents.

[070] In the present invention, the term “effective amount” means an amount that is high enough to provide a desired effect, but low enough to prevent serious side effects under medical evaluation. The amount of microorganisms administered to the body by the composition of the present invention can be appropriately adjusted considering the route of administration and the target of administration.

[071] The composition of the present invention can be administered to an individual one or more times per day. Unit dosage means physically discrete units suitable for unit administration to human individuals and other mammals, and each unit comprises a suitable pharmaceutical carrier and a predetermined amount of the Lactobacillus crispatus KBL693 strain of the present invention to provide 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 dosage of the composition of the present invention is 0.001 g ± 10 g, and preferably 0.01 g ± 5 g per dose. Petition 870260058402, dated 06 / 16 / 2026, page 29 / 106 20 / 42 The pharmaceutically effective amount of the microorganism of the present invention is 0.01 g ± 10 g / day. However, the dosage varies depending on the severity of the patient's disease and the effective microorganisms and auxiliary ingredients used in conjunction. Furthermore, the total daily dosage can be divided into several doses and administered continuously as needed. Consequently, the dosage ranges above do not limit the scope of the present invention in any way.

[072] Furthermore, the term “pharmaceutically acceptable”, as used above, refers to a composition that is physiologically acceptable and does not cause an allergic reaction, such as gastrointestinal upset, dizziness or a similar reaction when administered to a human being.

[073] A composition of the present invention can be formulated using methods known in the art so that rapid, sustained, or delayed release of the active ingredients, after administration to a mammal, can be provided. Dosage forms can be powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatin capsules, sterile injection solutions, or sterile powders. Furthermore, the composition of the present invention for preventing or treating allergic diseases, such as atopic dermatitis, and / or inflammatory or autoimmune diseases, can be administered by various routes, including oral, transdermal, subcutaneous, intravenous, or intramuscular administration. The dosage of the active ingredients can be appropriately selected depending on various factors, such as the route of administration, the patient's age, sex, weight, and severity.The composition of the present invention for the treatment of allergic diseases, such as atopic dermatitis, and / or inflammatory or autoimmune diseases, can be administered in combination with a known compound having the effect of preventing, relieving or treating the relevant symptoms.

[074] The pharmaceutical composition of the present invention, in particular, can be supplied in an oral unit dosage form of a formulation with Petition 870260058402, dated 06 / 16 / 2026, p. 30 / 106 21 / 42 Enteric coating. The term “enteric coating,” as used herein, comprises any known pharmaceutically acceptable coating that can remain in the stomach without being degraded by gastric acid and can disintegrate sufficiently in the intestinal tract to release active ingredients therein. The “enteric coating” of the present invention refers to a coating that can be maintained for 2 hours or more when an artificial gastric juice, such as a pH 1 HCl solution, comes into contact with it at 36°C to 38°C and subsequently can degrade, preferably in an artificial intestinal juice, such as a pH 6.8 KH2PO4 buffer solution, within 30 minutes.

[075] The enteric coating of the present invention is coated onto a core in an amount of about 16 mg to 30 mg, preferably 16 mg to 20 mg or 25 mg or less. When the thickness of the enteric coating of the present invention is 5 μm to 100 μm, and preferably 20 μm to 80 μm, satisfactory results can be obtained as an enteric coating. The enteric coating material can be suitably selected from known polymeric materials. Suitable polymeric materials are listed in a number of known documents [L. Lachman et al., The Theory and Practice of Industrial Pharmacy, 3rd ed., 1986, pp. 365–373; H. Sucker et al., Pharmazeutische Technologie, Thieme, 1991, pp. 355–359; Hagers Handbuchder pharmazeutischen Praxis, 4th ed., Vol. 7, p. 739 to 742 and 766 to 778, (SpringerVerlag, 1971); and Remington's Pharmaceutical Sciences, 13th ed., p. 1689 to 1691 (Mack Publ., Co.[ , 1970)], and cellulose ester derivatives, cellulose ethers, an acrylic resin-methylacrylate copolymer, and a copolymer of maleic acid and phthalic acid derivatives may be included.

[076] The enteric coating of the present invention can be prepared using a conventional enteric coating method in which an enteric coating solution is sprayed onto a core. Suitable solvents Petition 870260058402, dated 06 / 16 / 2026, page 31 / 106 The solvents used for the enteric coating process are alcohols such as ethanol, ketones such as acetone, halogenated hydrocarbon solvents such as dichloromethane (CH2Cl2), and mixed solvents of these solvents. A softener such as di-n-butyl phthalate or triacetin is added to the coating solution in a ratio of 1:approximately 0.05 to approximately 0.3 (coating material:softener). It is suitable to carry out the spraying process continuously, and the amount of spraying can be adjusted according to the coating conditions. The spray pressure can be adjusted in various ways, and satisfactory results can generally be obtained with a spray pressure of approximately 1 bar to approximately 1.5 bar.

[077] In another embodiment of the present invention, the present invention provides the use of said strain or said composition to prevent or treat allergic diseases, such as atopic dermatitis, and / or inflammatory or autoimmune diseases, and the use of said strain or said composition to prepare a therapeutic agent for the above diseases.

[078] Specifically, the present invention relates to a composition comprising at least one selected from the group consisting of said strain, cultures of said strain, lysates of said strain and extracts of said strain, for use in the prevention or treatment of allergic diseases including atopic dermatitis, and / or inflammatory or autoimmune diseases.

[079] The present invention also relates to the use of a composition to prepare a preventive or therapeutic drug for allergic diseases, including atopic dermatitis, and / or inflammatory or autoimmune diseases, comprising at least one selected from the group consisting of said strain, cultures of said strain, lysates of said strain and extracts of said strain.

[080] The term 'prevention', as used in this document, is associated with avoiding, delaying, stopping or preventing diseases or reducing them. Petition 870260058402, dated 06 / 16 / 2026, p. 32 / 106 23 / 42

[081] The term 'treatment', as used in this document, is associated with the care of an individual suffering from illnesses, in order to improve, cure or reduce the symptoms of the illnesses or reduce or stop the progression of the illnesses.

[082] In another embodiment of the present invention, the present invention provides a method for preventing or treating allergic diseases, including atopic dermatitis, and / or inflammatory or autoimmune diseases, comprising administering a pharmaceutically effective amount of the strain or said composition to an individual in need of prevention or treatment of said diseases, or in need of relief from said diseases.

[083] Specifically, the present invention provides a method for treating allergic diseases, including atopic dermatitis, and / or inflammatory or autoimmune diseases, comprising administering a therapeutically effective amount of at least one selected from the group consisting of said strain, cultures of said strain, lysates of said strain and extracts of said strain to an individual in need of treatment.

[084] Since the pharmaceutical composition used for the method to prevent or treat said diseases, and the method of administration thereof, have been described above, overlapping contents between the composition and the method will be omitted in this document to avoid undue complexity of the present specification.

[085] Meanwhile, the said individual to whom the composition for preventing or treating said diseases may be administered includes all animals, including human beings. For example, the individual may be an animal, such as a dog, cat or mouse.

[086] In another embodiment of the present invention, the present invention relates to a cosmetic composition comprising a pharmaceutically effective amount of at least one component selected from the group consisting of the component Petition 870260058402, dated 06 / 16 / 2026, p. 33 / 106 24 / 42 cellular strain of Lactobacillus crispatus KBL693, cultures of said strain, lysates of said strain, and extracts of said strain.

[087] A cosmetic composition may be characterized by its function of relieving at least one sensitive skin condition selected from the group consisting of skin allergy, urticaria, atopic dermatitis, psoriasis, fungal infections and eczema, but not limited to these.

[088] The term 'cosmetic composition', as used herein, refers to a composition comprising at least one selected from the group consisting of the cellular component of the Lactobacillus crispatus KBL693 strain, cultures of said strain, lysates of said strain and extracts of said strain, and may assume any type of dosage form. For example, cosmetics prepared using said cosmetic composition may include creams, sachets, lotions, essences, toners, bases and makeup bases, and may be marketed in any of the dosage forms listed above to achieve the objective of the present invention, but not limited to these. The ingredients comprised in the cosmetic composition of the present invention include those commonly used in cosmetic compositions, in addition to the above ingredients, for example, conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances, and carriers.

[089] In another embodiment of the present invention, the present invention relates to a functional plaster comprising at least one selected from the group consisting of the cellular component of the Lactobacillus crispatus KBL693 strain, cultures of said strain, lysates of said strain and extracts of said strain, and said functional plaster can be used for cosmetic or medical purposes.

[090] The plaster can be characterized by its function of relieving at least one sensitive skin condition selected from the group consisting of skin allergy, skin urticaria, atopic dermatitis, psoriasis, fungal infections and Petition 870260058402, dated 06 / 16 / 2026, page 34 / 106 25 / 42 eczema, but not limited to these.

[091] In the present invention, the patch is typically a small adhesive bandage containing the substances to be delivered, and the bandage may assume a variety of forms. The simplest form is a single patch body comprising a reservoir containing the substances to be delivered over a support. The reservoir is typically formed from a cosmetically or pharmaceutically acceptable pressure-sensitive adhesive, but in some cases it may also be formed from non-adhesive materials provided with a thin adhesive layer suitable for the skin contact surface. The rate at which the substances to be delivered are administered from the patch to the individual using the patch may be altered because the permeability of the skin to the substances to be delivered generally depends on the individual and the location of the skin, which may be easily selected by those skilled in the art.

[092] The present invention will now be described in more detail by means of examples. These examples are only for illustrative purposes of the present invention, and it will be evident to those skilled in the art that the scope of the present invention should not be interpreted as being limited by these examples.

[093] Example 1. Relief and treatment effects of KBL693 for allergic reactions resulting from inhibition of histamine secretion in basophils

[094] It has been reported that, in an allergic reaction, histamine is expressed in tissues, causing an inflammatory reaction, and suppression of histamine secretion leads to relief of allergic symptoms by blocking the histamine reaction. The present invention attempted to select probiotic strains that may exhibit effects in relieving and treating allergic reactions by inhibiting histamine secretion. To this end, a total of eight Lactobacillus strains derived from the human vagina were evaluated for their ability to suppress histamine secretion. The ability to suppress histamine secretion was confirmed by inducing Petition 870260058402, dated 06 / 16 / 2026, page 35 / 106 26 / 42 degranulation after culture of the RBL-2H3 cell line and then measurement of the activity of β-hexosaminidase co-secreted with histamine, using a colorimetric reaction with a substrate. 1-1. Incubation of RBL-2H3 cell lines

[095] RBL-2H3 cells (ATCC NO. CRL-2256) were cultured in DMEM medium supplemented with 10% FBS (fetal bovine serum), penicillin (100 μg / mL), and streptomycin (100 μg / mL) at 37°C under 5% CO2, and then subcultured once every three days. RBL-2H3 cells were seeded in a 6-well plate with a concentration of 1 χ¹⁰⁶ cells / well, then cultured for 3 hours and subsequently treated with IgE (0.5 μg / mL) to incubate for 16–20 hours. 1-2. Incubation of strains and preparation of the culture solution

[096] The Lactobacillus crispatus strains to be used were cultured in MRS medium supplemented with 0.5% cysteine, activated by means of a total of two subcultures at 18-hour intervals, and then used in experiments. The resulting culture solution was centrifuged at 15,000 χ g for 3 minutes to collect the supernatant. 1-3. Inducing degranulation after treatment of strains

[097] After removing the medium from RBL-2H3 cells that had been previously seeded in a 6-well plate, the cells were washed twice with 1 mL of Siraganian buffer (pH 7.2). Subsequently, the cells were treated with 120 μL of the previously prepared bacterial culture solution or the positive control group ketotifen (20 μg / mL) per well, incubated for 20 minutes in a 5% CO2 incubator at 37°C, followed by treatment with 60 μL of antigen (DNP-HAS, 1 μg / mL) to induce degranulation by the antigen-antibody reaction. As a negative control, degranulation was induced after treatment with 120 μL of Siraganian buffer. The reaction was carried out in a 5% CO2 incubator at 37°C for 20 minutes, and then the supernatant was collected. Petition 870260058402, dated 06 / 16 / 2026, page 36 / 106 27 / 42 -4. Identifying the color reaction

[098] To identify β-hexosaminidase activity, 25 μL of the supernatant collected in Example 1-3 were transferred to each well of a 96-well plate, and then 25 μL of each substrate (p-nitrophenyl N-acetyl-D-glucosaminidase) were added, followed by a reaction at 37°C in a 5% CO2 incubator for 90 minutes. Then, 200 μL of stopping solution (Na2CO3 / NaHCO3) were added to stop the reaction, and the absorbance was measured at 405 nm. The absorbance when treated with each type of Lactobacillus crispatus was compared to the negative control group and converted to a percentage to show the level of degranulation inhibition.

[099] As a result, as can be seen in Figure 1 and Table 1, ten types of Lactobacillus crispatus strains, including strain KBL693, showed significantly lower amounts of β-hexosaminidase secretion compared to the negative control group, and nine types showed even lower amounts of β-hexosaminidase secretion than the commercially available antihistamine ketotifen, a positive control. As a result, Lactobacillus crispatus strains, including strain KBL693, were found to exhibit excellent degranulation prevention activity to effectively alleviate allergic symptoms caused by excessive histamine secretion. Table 1 Species Strain under treatment or treatment drug Β-hexosaminidase (modulation) - (Negative control) Siraganian buffer 1.0 - (Positive control) Ketotifen 0.2569 Lactobacillus crispatus KBL693 0.2140 Lactobacillus crispatus KBL694 0.2063 Lactobacillus crispatus KBL692 0.1904 Lactobacillus crispatus KBL702 0.2065 Lactobacillus crispatus KBL706 0.1943 Lactobacillus crispatus KBL707 0.2008 Petition 870260058402, dated 06 / 16 / 2026, p. 37 / 106 28 / 42 Lactobacillus crispatus KBL709 0.2128 Lactobacillus crispatus KBL696 0.2331 Lactobacillus crispatus KBL708 0.2464 Lactobacillus crispatus SNUV272-1 0.3632 Lactobacillus fermentum SNUV417 0.9284 Lactobacillus jensenii SNUV221 0.7088 Lactobacillus vaginalis SNUV351 1.0336

[0100] Example 2. Relief and treatment effects of KBL693 for allergic reactions resulting from inhibition of Th2-type cytokines in T cells

[0101] It has been reported that cytokines related to type 2 helper T cells (Th2), such as IL-4 and IL-5, contribute to chronic allergic reactions by increasing Th2-related immune responses and increasing IgE production (Passante E, Inflamm. Res. 2009). The present invention further attempted to compare the effects on the inhibition of chronic allergic reactions between Lactobacillus crispatus strains showing an excellent effect on the inhibition of histamine secretion in Example 1. To this end, the effect of inhibiting IL-4 and IL-5 secretion was tested as follows, using the EL4 cell line, which is a mouse T cell line. 2-1. Incubation of EL-4 cell lines

[0102] EL4 cells (ATCC NO. TIB-39) were cultured in DMEM medium supplemented with 10% FBS, penicillin (100 μg / mL) and streptomycin (100 μg / mL) at 37°C under 5% CO2, and then subcultured once every three days. EL4 cells were seeded in a 24-well plate at a concentration of 4 x 10⁵ cells / well, then cultured for 16–20 hours to induce allergic reactions, and treated with the strains. 2-2. Incubation of strains and recovery

[0103] A total of nine Lactobacillus crispatus strains, including KBL693 (KBL693, KBL709, KBL702, KBL696, KBL708, KBL707, KBL692, KBL694, and KBL706), were cultured in MRS medium supplemented with 0.5% cysteine ​​and activated through a total of two subcultures at 18-minute intervals. Petition 870260058402, dated 06 / 16 / 2026, p. 38 / 106 The resulting culture solution was centrifuged at 15,000 χ²g for 3 minutes, and the precipitate was washed with PBS (Phosphate-Buffered Saline) buffer. The strains were stained for 15 minutes with SYTO9 and PI using the LIVE / DEAD™ BacLight™ Bacterial Count and Viability Kit for flow cytometry (Thermo Fisher Scientific, USA). Then, the contained microspheres were added, and the number of live stained bacteria was calculated using a flow cytometry assay. 2-3. Treatment of strains and measurement of IL-4 and IL-5 secretion levels after inducing allergic reactions.

[0104] In order to induce allergic reactions in EL4 cells previously seeded in a 24-well plate, each well was treated with 100 μL of PMA (20 ng / mL) and ionomycin (1 μg / mL). Then, 300 μL of the previously prepared strains were treated at a cell-to-strain ratio of 1:10. After incubation for 24 hours in a 5% CO2 incubator at 37°C, the supernatant was collected, and the mouse IL-4 ELISA set (Cat NO. 555232, BD OptEIA™) and the mouse IL-5 ELISA set (Cat NO. 555236, BD OptEIA™) were used to measure the amount of secreted IL-4 and IL-5, according to the manufacturer's method.

[0105] As a result, as shown in Figure 2 and Figure 3, those Lactobacillus crispatus strains generally showed effects in inhibiting the secretion of IL-4 and IL-5, and especially in the KBL693 strain, among those, the lowest amount of IL-4 secretion was found, and the second lowest amount of IL-5 secretion was found. Consequently, it was discovered that KBL693 can provide therapeutic and preventive effects in allergies by inhibiting the secretion of Th2-type cytokines that mediate an allergic reaction.

[0106] Example 3. Effects of KBL693 in the relief and treatment of allergic reactions resulting from IgE inhibition

[0107] Immunoglobulin E (IgE) is a type of immunoglobulin known for Petition 870260058402, dated 06 / 16 / 2026, p. 39 / 106 30 / 42 being involved in the development of allergic diseases. In general, the total amount of IgE in serum is measured as one of the most prioritized methods for diagnosing an allergic disease. Thus, the present invention attempted to verify the inhibitory effect of IgE secretion by KBL693 using the U266B1 cell line, which is a human B cell line. 3-1. Incubation of U266B1 cell lines

[0108] U266B1 cells (ATCC NO. TIB-196) were cultured in RPMI1640 medium supplemented with 10% FBS, penicillin (100 μg / mL), and streptomycin (100 μg / mL) at 37°C under 5% CO2, and then subcultured once every three days. U266B1 cells were seeded in a 24-well plate at a concentration of 1 x 10⁵ cells / well, then cultured for 16–20 hours to induce inflammatory reactions, and then treated with the strains. 3-2. Incubation of strains and recovery

[0109] The strains to be used in this experiment were prepared in the same manner as in Example 2-2. 3-3. Treatment of strains and measurement of IgE secretion levels after inducing allergic reactions.

[0110] In order to induce allergic reactions in U266B1 cells previously seeded in a 24-well plate, each well was treated with 100 μL of LPS (10 μg / mL) and IL-4 (5 ng / mL). Then, 300 μL of the previously prepared strains were treated at a cell-to-strain ratio of 1:10. After incubation for 48 hours in a 5% CO2 incubator at 37°C, the supernatant was collected, and the human IgE ELISA kit (Cat. No. E88-108, Bethyl Laboratories) was used for measurement, according to the manufacturer's method.

[0111] As a result, as shown in Figure 4, it was confirmed that the groups treated with seven strains of Lactobacillus crispatus, including KBL693, showed effects in inhibiting IgE secretion, compared to the group of Petition 870260058402, dated 06 / 16 / 2026, page 40 / 106 31 / 42 negative control. The present experiment showed that KBL693 can provide therapeutic and preventive effects in allergic diseases by inhibiting IgE, one of the main factors involved in allergic reactions.

[0112] Example 4. Analysis of immunoregulatory and inflammation-inhibiting effects of KBL693

[0113] The immunoregulatory and inflammation-inhibiting effects of KBL693 were also verified, in addition to its antiallergic efficacy. To this end, the relationship between IL-10, a representative cytokine that has an immunoregulatory function, and TNF-α and IL-6, which are the cytokines that are the main indicators of an inflammatory reaction (IL-10 / TNF-α, IL-10 / IL-6), was measured using macrophages, which play a crucial role in the inflammatory reaction. 4-1. Incubation of RAW264.7 cell lines

[0114] RAW264.7 cells (ATCC NO. TIB-71) were cultured in DMEM medium supplemented with 10% FBS, penicillin (100 μg / mL), and streptomycin (100 μg / mL) at 37°C under 5% CO2, and then subcultured once every three days. The incubated RAW264.7 cells were seeded in a 24-well plate at a concentration of 1 χ¹⁰⁵ cells / well, then cultured for 16–20 hours, and subsequently used for Example 4–3. 4-2. Incubation of strains and sample preparation

[0115] The culture solutions were corrected with the same number of live bacteria and then prepared as three samples: live bacteria, pasteurization, and heat death. The samples for live bacteria, pasteurization, and heat death were prepared according to the same method as Example 2-2.

[0116] Next, the sample for pasteurization was reacted at 70°C for 30 minutes and prepared by the same centrifugation process. The heat-kill sample was sterilized at 121°C for 15 minutes and prepared using the same process. As a negative control group, MRS medium supplemented with 0.5% cysteine ​​was used. Petition 870260058402, dated 06 / 16 / 2026, page 41 / 106 32 / 42 was used. 4-3. Treatment of strains and measurement of the amount of inflammatory cytokines secreted after inducing inflammatory reactions.

[0117] In order to induce inflammatory reactions in RAW264.7 cells previously seeded in a 24-well plate, each well was treated with 500 μL of LPS (20 ng / mL). Next, 300 μL of the previously prepared strains were treated at a cell-to-strain ratio of 1:10 and incubated for 24 hours in a 5% CO2 incubator at 37°C. The supernatant in the cultured cell-strain mixture was collected, and Mouse TNF ELISA Set II (Cat No. 558534, BD OptEIA™), Mouse IL-6 ELISA Set (Cat No. 555240, BD OptEIA™), and Mouse IL-10 ELISA Set (Cat No. 555252, BD OptEIA™) were used to measure the quantity of each cytokine, according to the manufacturer's method.

[0118] As a result, as shown in Figure 5, it was confirmed that KBL693, among the Lactobacillus crispatus strains, specifically induced IL-10 secretion and therefore showed excellent effects in terms of immunoregulation and suppression of the inflammatory reaction. As shown in Figures 6 and 7, based on the ability to secrete IL-10 corrected with the pro-inflammatory cytokines TNF-α and IL-6, it was confirmed that the KBL693 treatment group showed a significantly improved inflammation-inhibiting effect and immunoregulatory capacity compared to the control group. Consequently, it was found that KBL693 has immunoregulatory and inflammation-inhibiting activities through IL-10 secretion.Furthermore, these effects were similarly confirmed not only in live bacteria, but also in pasteurized or heat-killed dead bodies (see Table 2 and Table 3), indicating that KBL693 can be used to regulate immune reactions and suppress inflammatory reactions in various forms, such as dead bacteria. Petition 870260058402, dated 06 / 16 / 2026, page 42 / 106 33 / 42 Table 2 IL-10 / TNF-a Live Heat-dead Pasteurized Control -1.7574022 (ND) -1.6268471 (ND) -1.9978721 (ND) KBL693 4.22079378 3.82025311 4.97474621 ND: Not Determined Table 3 IL-10 / IL-6 Live Heat-dead Pasteurized Control -3.5789041 (ND) -4.2268549 (ND) -4.712413 (ND) KBL693 4.95082596 4.9303055 6.46882207 ND: Not Determined

[0119] Example 5. Analysis of the antifungal effects of KBL693

[0120] The antifungal effect of KBL693 was confirmed by a spot test method. Approximately 1% of KBL693 was inoculated into MRS liquid medium and then incubated in an incubator at 37°C for approximately 24 hours under anaerobic conditions for stationary culture. The culture solution in which the cells were incubated was stained onto MRS solid medium, which was prepared under anaerobic conditions, in a quantity of 10 μL at a time, and then incubated at 37°C under anaerobic conditions for approximately 24 hours. Malassezia furfur KCTC 7545, a fungal microorganism, was prepared by inoculating it into mYPG liquid medium, which was prepared under aerobic conditions, at a rate of 1% and followed by incubation in an incubator at 37°C for approximately 24 to 48 hours. The soft mYPG medium to be used for antifungal efficacy evaluation was prepared with the components shown in Table 4, and 2.5 mL of the prepared culture medium were inoculated with the culture solution in which 500 μL of M. furfur were incubated.2.5 mL of mYPG medium inoculated with M. furfur were poured onto MRS medium stained with KBL693 and dried for 1 hour. The dried medium was incubated under aerobic conditions in an incubator at 37°C for 24 to 48 hours. When a clear zone was identified in the culture medium, antifungal activity was determined by measuring the length of the outer side of the bacteria. Petition 870260058402, dated 06 / 16 / 2026, p. 43 / 106 34 / 42 of lactic acid stained up to the clear zone. The clear zone is a part where fungal growth was inhibited, and the antifungal activity caused by lactic acid bacteria was determined along the length up to the clear zone.

[0121] As a result of three repeated experiments, clear zones were identified at intervals of approximately 4.33 mm in the area stained with KBL693 (Figure 8), indicating that KBL693 can effectively suppress the growth of fungal microorganisms. Table 4 Ingredients Proportion (g / 1) Malt extract 5.0 Peptone 10.0 Glucose 20.0 Tween 40 1.0 Tween 80 1.0 Agar 4.5

[0122] Example 6. Effects of KBL693 in relieving atopic conditions

[0123] Based on a comprehensive review of the in vitro screening test results, animal tests were conducted selecting Lactobacillus crispatus KBL692, KBL693, and KBL702, which had the most excellent immunoregulatory effect. In order to verify the effect on atopic relief among the allergy-enhancing effects of KBL693, the NC / Nga mouse model, an animal model of atopic skin disease, was used.

[0124] After dividing the NC / Nga mice into groups of five mice, the back of each mouse was shaved from the lower ear to the upper tail and the mice were left for 24 hours. Then, 200 μL of 1% DNCB (dinitrochlorobenzene) solution (acetone: olive oil = 3:1) was applied twice weekly to the shaved area to induce atopic dermatitis. From the third week of dermatitis induction, 200 μL of PBS was administered orally to the mice in the control group daily; the strain Petition 870260058402, dated 06 / 16 / 2026, p. 44 / 106 35 / 42 of the cultured test sample was centrifuged, washed by dilution with PBS and recovery, and then prepared so that at least 2 χ¹⁰⁹ CFU could be added to 200 μL of PBS, which was administered orally to mice in the test group at 200 μL / day. Meanwhile, 200 μL of dexamethasone (60 μg / mL) was administered to mice in the positive control group. Then, during three weeks of bacterial administration, the dermatitis scores of the mice in the control and test groups were measured weekly, and in the 3rd week after bacterial administration, the scratch time and skin thickness of the mouse were measured. 6-1. Dermatitis score assessment

[0125] To assess DNCB-induced skin lesions, the dermatitis score was measured using the following method. Skin conditions were monitored by taking photographs for 3 weeks at one-week intervals starting from the 3rd week after the strain was administered. Four indicators of dryness, edema, erythema / hemorrhage, and skin erosion / excoriation were checked. A condition without lesions was classified as 0 points, a mild condition as 1 point, a moderate condition as 2 points, and a severe condition as 3 points, and the total score was evaluated.

[0126] As a result, as shown in Figure 9, the DNCB-induced dermatitis score was significantly reduced in the group dosed with KBL693, compared to the control (negative) group where atopic dermatitis was induced and the groups dosed with other Lactobacillus crispatus strains. As a result, the treatment effect of atopic dermatitis according to the administration of KBL693 was verified. 6-2. Itch-relieving effects

[0127] In order to verify the itch-relieving effect of administering KBL693 in mouse models suffering from dermatitis Petition 870260058402, dated 06 / 16 / 2026, p. 45 / 106 In the DNCB-induced atopic dermatitis study (36 / 42), the itch time was measured using a video recording of the mouse models for 10 minutes after 3 weeks of strain administration.

[0128] As a result, as can be seen in Figure 10, it appears that the itching time was significantly reduced in the test group in which KBL693 was administered, compared with the control (negative) group in which atopic dermatitis was induced and the groups dosed with other strains of Lactobacillus crispatus, which confirmed that the itching symptoms of atopic dermatitis were quite relieved with the administration of KBL693. 6-3. Decrease in skin thickness

[0129] In order to verify the itch-relieving effect after administration of KBL693 to mouse models suffering from DNCB-induced atopic dermatitis, ear thickness and dorsal skin thickness of the mice were measured with calipers three weeks after the strain was administered, and relief of the edema symptom due to atopic dermatitis was observed.

[0130] As a result, as can be seen in Figures 11 and 12, it was observed that in the test group dosed with KBL693 and in the positive control group, the thicknesses of the ear and dorsal skin were significantly reduced. In Figures 11 and 12, 693-7, 693-8 and 693-9, respectively, signify the test groups in which KBL693 was prepared so that at least 2 χ 107, 2 χ 108 and 2 χ 109 CFU could be added to 200 μL of PBS, which were administered orally to the mice in the test group at 200 μL / day.

[0131] Example 7. Effects of KBL693 on the treatment and prevention of asthma in mouse models of ovalbumin (OVA)-induced asthma.

[0132] In order to verify the effect of KBL693 (LC206) on allergic asthma, mouse models of OVA-induced asthma were prepared as follows, a histopathological examination was conducted and the expression level of IL-5 and IL-15 was evaluated. 870260058402, 16 / 06 / 2026, p. 46 / 106 37 / 42 13. 7-1. Preparation of mouse models of OVA-induced asthma

[0133] Five-week-old Balb / c mice with an average weight of about 15 g were acclimatized for one week, and then mice that showed no abnormalities on basic physical examination were selected. As shown in Figure 13 (A), the mice to be used were classified into a normal control group (Control-PBS; no OVA administered or inhaled; n = 8), an asthma-induced control group (OVA-PBS; OVA administered or inhaled; n = 9), and a test group (OVA-LC206; n = 8). Before the mice were sacrificed on day 31 from the start of the experiment, 200 μL of PBS were administered orally to the normal control group and the asthma-induced control group daily, and 200 μL of KBL693 diluted in PBS to at least 2 χ 109CFU / 0.2 mL were administered orally to the test group daily.

[0134] Sensitization was performed by intraperitoneal injection of 200 μL of a phosphate buffer (pH 7.4) in which 2 mg of aluminum hydroxide (Imject™ Alum Adjuvant, Thermofisher) and 20 μg of OVA (Ovalbumin, Sigma-Aldrich) were discontinued, to mice one week later (D7) and three weeks later (D21) after acclimation for one week, respectively. From day 28 to day 30 from the start of the experiment, 1% OVA was inhaled into the lungs via intranasal administration (three provocations). Pentobarbital was administered 24 hours after the last provocation (D31), and then a bronchial incision was made to collect lung tissue samples. 7-2. Histopathological Examination

[0135] Infiltration of inflammatory cells consisting of eosinophils, neutrophils, and macrophages can be observed in the bronchi where the antigen is treated. Thus, in order to verify the effect of KBL693 on asthma, an examination was performed. Petition 870260058402, dated 06 / 16 / 2026, page 47 / 106 38 / 42 Histopathological examination of lung tissue samples from each group obtained in Example 71. The lung tissue samples obtained in Example 7-1 were prepared as 4 μm thick tissue sections using a conventional formalin-fixed paraffin-embedded (FFPE) tissue section procedure and then H&E staining was conducted with hematoxylin and Eosin Y (Eosin Y; ThermoShandon, Pittsburgh, PA). After H&E staining, tissue sections from each group were observed with a light microscope (Figure 13(B)).

[0136] As a result, as shown in Figure 13 (B), in the control group of induced asthma (OVA-PBS), many inflammatory cells, including eosinophils, were infiltrated around the bronchioles, and hyperproliferated epithelial cells and thickened bronchial smooth muscle were also found. On the other hand, in the test group dosed with KBL693 (OVA-LC206), the infiltration of inflammatory cells including eosinophils was significantly reduced, the thickness of the bronchial tissue was reduced, and the epithelial cells were hardly damaged. As a result, the therapeutic effect of KBL693 in OVA-induced allergic asthma was confirmed. 7-3. Total IL-5 and total IL-13 expression level assay

[0137] A flow cytometer (LSRFortessa X-20, BD) was used to assess immunocytes in lungs from lung tissue samples collected from each group. To stain CD4+ IL-5+ T cells, CD4+ IL-13+ T cells, antibodies to various markers (CD45 Anti-Mouse, BioLegend; CD3ε Anti-Mouse, BD; IL-5 Anti-Mouse / Human, BioLegend; IL-13 Anti-Mouse, Invitrogen) were used. IL-5+ and IL-13+ cells were determined by counting IL-5 or IL-13-producing cells among lymphocytes having CD45 and CDε as markers (Figure 14).

[0138] As a result, as shown in Figure 14, it was found that the levels of IL-5 and IL-13 in the asthma-induced control group mice Petition 870260058402, dated 06 / 16 / 2026, page 48 / 106 39 / 42 (OVA-PBS) significantly increased compared to the normal control group (PBS), while total IL-5 and total IL-13 levels in mice in the KBL693-dosed test group (OVA-KBL693) were significantly decreased compared to the asthma-induced control group. Consequently, KBL693 of the present invention was found to provide therapeutic and preventive effects in allergic asthma by inhibiting the secretion of IL-5 and IL-13, which are Th2-type cytokines that mediate allergic reactions.

[0139] Example 8. Effects of KBL693 on the treatment and prevention of asthma in house dust mite-induced asthma (HDM) models

[0140] House dust mites are the allergen that is the main cause of extrinsic asthma. In order to verify the effect of KBL693 on allergic asthma, mouse models with HDM-induced asthma were prepared as follows, airway hypersensitivity was assessed, and the ratio of eosinophils to CD45+ cells, the ratio of CD4 IL-5+ T cells to CD4+ T cells, and the ratio of CD4 IL-13+ T cells to CD4+ T cells were determined.

[0141] 8-1. Preparation of mouse models with HDM-induced asthma

[0142] Five-week-old Balb / c mice with an average weight of approximately 15 g were acclimatized for one week, and then mice that showed no abnormalities on basic physical examination were selected. The mice to be used were classified into a normal control group (Control-PBS; no HDM administered), an asthma-induced control group (HDM-PBS; HDM administered), and a test group (HDM-KBL693), and five mice from each group were used to assess airway hyperresponsiveness (AHR), and eight mice from each group were used to assess lung immunocytes.

[0143] Sensitization was performed by intranasal administration of 50 μL of Petition 870260058402, dated 06 / 16 / 2026, page 49 / 106 40 / 42 mice were given a phosphate buffer (pH 7.4) in which 10 μg of HDM (Greer) was suspended, one week after (D7) acclimation for one week. One week after sensitization, 50 μL of a phosphate buffer in which 10 μg of HDM was suspended was inhaled into the lungs by intranasal administration for five days (D14 ~ D18) (five provocations). Pentobarbital was administered 24 hours after the last provocation (D19), and then airway hyperresponsiveness (AHR) was assessed (Example 8-2), and a bronchial incision was made to collect lung tissue samples. 8-2. AHR Assessment

[0144] A mouse anesthetized by administration of pentobarbital was connected to an AHR (FinePoint Resistance and Compliance, DSIBuxco) meter, and various concentrations of methacholine dissolved in PBS (0, 5, 10, 20, and 40 mg / mL) were administered. The AHR value was then calculated by measuring the volume of air passing through the airways (Figure 15).

[0145] As a result, as shown in Figure 15, it was confirmed that as the methacholine concentration increased, the AHR (Rl) increased slowly in the normal control group (CTRL), while the AHR increased rapidly in the asthma-induced control group (HDM). On the other hand, in the test group dosed with KBL693 (HDM + KBL693), the AHR was significantly reduced compared to the asthma-induced group (HDM) at all methacholine concentrations. This difference was more obvious when administered with high methacholine concentrations than with low methacholine concentrations. Consequently, it was found that KBL693 can effectively inhibit asthma-causing AHR and therefore can be used effectively for the treatment and prevention of allergic asthma.

[0146] 8-3. Determination of the proportion of eosinophils in CD45+ cells, the proportion of IL-5+ CD4 T cells in CD4+ T cells and the proportion of CD4 T cells Petition 870260058402, dated 06 / 16 / 2026, page 50 / 106 41 / 42 IL-13+ on CD4+ T cells

[0147] A flow cytometer (LSRFortessa X-20, BD) was used to assess immunocytes in the lungs. To stain eosinophils and CD4+IL-5+ T cells, CD4+IL-13+ T cells, antibodies to various markers (CD45 Anti-Mouse, BioLegend; Siglec-F Anti-Mouse, BD; CD11b Anti-Mouse, BD; CD3ε Anti-Mouse, BD; TCRβ Anti-Mouse, BioLegend; CD4 Anti-Mouse, BioLegend; IL-5 Anti-Mouse / Human, BioLegend; IL-13 Anti-Mouse, Invitrogen) were used. Eosinophils were determined by counting Siglec-f+CD11b+ cells, among cells expressing CD45, a common leukocyte marker, and CD4+IL-5+ T cells. CD4+IL-13+ T cells were determined by counting IL-5 or IL-13 producing cells, among CD4+ T cells having CDε, TCRβ, and CD4 as markers (Figure 16).

[0148] As a result, as shown in Figure 16, it was confirmed that, in the case of the asthma-induced control group (HDM), the proportions of immunocytes, i.e., eosinophils, CD4 IL-5+ T cells, and CD4 IL-13+ T cells, were all significantly increased compared to the normal control group (PBS), while in the case of the test group dosed with KBL693 (HDM + KBL693), the proportions of eosinophils, CD4 IL-5+ T cells, and CD4 IL-13+ T cells were all significantly reduced compared to the asthma-induced control group. Consequently, it was found that KBL693 can provide therapeutic and preventive effects in allergic asthma by inhibiting inflammatory cells, i.e., eosinophils, CD4 IL-5+ T cells, and CD4 IL-13+ T cells.

[0149] Specific aspects of the present invention have been described in detail above, and it is obvious to those skilled in the art that these specific aspects are only preferred embodiments, and the scope of the present invention is not limited by them. Therefore, the scope of the present invention is substantially defined by the following claims, with equivalents to the claims Petition 870260058402, dated 06 / 16 / 2026, page 51 / 106 42 / 42

[0150] Name of the depository organization: Korea Bioscience & Biotechnology Research Institute Access Number: KCTC13519BP Access Date: 20180427 Industrial Applicability

[0151] The Lactobacillus crispatus KBL693 strain (KCTC Accession No. 13519BP) according to the present invention attenuates allergic cell reactions, significantly improves the symptoms of atopic dermatitis, and exhibits anti-inflammatory, immunoregulatory, and antifungal effects. Thus, the single strain alone can achieve all the objectives of alleviating allergic diseases and improving inflammatory and autoimmune diseases, thus finding advantageous applications as a probiotic substance. Furthermore, the strain, based on its antifungal activity, can be advantageously used in an external skin preparation against various skin diseases caused by fungi, and in a cosmetic composition and a functional plaster for the relief of sensitive skin. Free Text from Sequence Listing Attached electronic file Petition 870260058402, dated 06 / 16 / 2026, page 52 / 106

Claims

1 / 2 CLAIMS 1. Feed composition presented in enteric-coated dosage form, CHARACTERIZED in that it comprises (i) at least one selected from the group consisting of the Lactobacillus crispatus KBL693 strain with KCTC Accession No. 13519BP, cultures of said strain and lysates of said strain and (ii) phosphate-buffered saline (PBS) solution in an enteric-coated core.

2. Antifungal composition presented in enteric-coated dosage form CHARACTERIZED in that it comprises (i) at least one selected from the group consisting of the Lactobacillus crispatus KBL693 strain with KCTC Accession No. 13519BP, cultures of said strain and lysates of said strain and (ii) PBS in an enteric-coated core.

3. Pharmaceutical composition for the treatment of allergic diseases, inflammatory diseases or autoimmune diseases presented in enteric-coated dosage form, CHARACTERIZED in that it comprises (i) at least one selected from the group consisting of the Lactobacillus crispatus KBL693 strain with KCTC Accession No. 13519BP, cultures of said strain and lysates of said strain and (ii) PBS in an enteric-coated core.

4. Pharmaceutical composition, according to claim 3, CHARACTERIZED in that said allergic diseases are selected from the group consisting of eczema, allergic asthma, allergic rhinitis, atopic dermatitis, allergic conjunctivitis and food allergy.

5. Pharmaceutical composition, according to claim 3, CHARACTERIZED in that said inflammatory or autoimmune diseases are selected from the group consisting of rheumatoid arthritis, rheumatic fever, lupus, systemic scleroderma, atopic dermatitis, psoriasis, psoriatic arthritis, asthma, Guillain-Barré syndrome, myasthenia gravis, dermatomyositis, polymyositis, sclerosis. Petition 870260058402, dated 06 / 16 / 2026, page 1.53 / 106 2 / 2 multiple, autoimmune encephalomyelitis, polyarteritis nodosa, Hashimoto's thyroiditis, temporal arteritis, juvenile diabetes, alopecia areata, pemphigus, aphthous stomatitis, autoimmune hemolytic anemia, Wegener's granulomatosis, Sjögren's syndrome, Addison's disease, Crohn's disease, Behçet's disease, edema, conjunctivitis, periodontitis, rhinitis, otitis media, chronic sinusitis, pharyngolaryngitis, tonsillitis, bronchitis, pneumonia, gastric ulcer, gastritis, colitis, gout, eczema, acne, contact dermatitis, seborrheic dermatitis, ankylosing spondylitis, fibromyalgia, osteoarthritis, shoulder periarthritis, tendinitis, tenosynovitis, myositis, hepatitis, cystitis, nephritis, sepsis, vasculitis, and bursitis. Petition 870260058402, dated 06 / 16 / 2026, pp. 54 / 106.

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