Novel lacticaseibacillus rhamnosus strain and use thereof

Abstract

The present invention relates to a novel Lacticaseibacillus rhamnosus HLL-002 strain exhibiting anti-inflammatory and anti-allergic activities and, more specifically, provides: a Lacticaseibacillus rhamnosus HLL-002 strain; a probiotic composition comprising same; a health functional food composition comprising the composition; and a pharmaceutical composition for preventing or treating allergic diseases, comprising the strain.

Description

Novel Lacticazeibacillus rhamnosus strain and its use

[0001] This application claims priority to Republic of Korea Patent Application No. 10-2024-0092018, filed July 11, 2024, the entire disclosure of which is incorporated herein by reference.

[0002] The present invention relates to a novel Lacticazei Bacillus rhamnosus HLL-002 strain exhibiting anti-inflammatory and anti-allergic activity, and more specifically, to a Lacticazei Bacillus rhamnosus HLL-002 strain; a probiotic composition comprising the same; a health functional food composition comprising the composition; and a pharmaceutical composition for preventing or treating allergic diseases comprising the strain.

[0003] Probiotics are microorganisms with antimicrobial and enzymatic activity that help maintain intestinal microbial balance, as well as their products. Furthermore, probiotics are defined as single or complex strains of live bacteria that are administered to humans or animals in the form of dried cells or fermented products to improve intestinal flora.

[0004] Examples of such probiotics include Bacillus species (Bacillus sp.) with excellent digestive enzyme production capabilities (amylase, protease, lipase, cellulase, phosphatase), Lacticase Bacillus species that produce lactic acid, and photosynthetic bacteria that metabolize odor-causing substances (ammonia, hydrogen sulfide, amines, etc.) remaining in livestock feces to prevent odor production.

[0005] Bacillus and Lacticazei Bacillus species, in particular, are known as highly useful probiotics because strains produce a variety of antimicrobial substances. These lactic acid bacteria produce antimicrobial peptides called bacteriocins, which have antimicrobial mechanisms unrelated to antibiotic resistance. Bacteriocins exhibit polymorphic characteristics, significantly varying in molecular weight, biochemical properties, and antimicrobial spectrum and mechanisms against hosts.

[0006] Meanwhile, allergy is a biochemical phenomenon that exhibits a specific and altered hypersensitivity reaction to a foreign substance (antigen, allergen). When the foreign substance causes symptoms, it is called an allergen, and the disease that results is called an allergic disease. Allergy is a pathological process of the body that appears as a result of the antigen-antibody reaction that is the cause of the occurrence, and is generally classified into four types based on the time required to cause the reaction and the type of complement involvement.

[0007] Among allergic diseases, atopic dermatitis (AD) is a chronic, recurrent skin inflammatory disease that occurs in approximately 20% of newborns and children in Korea and can persist into adulthood. Atopic dermatitis is an inflammatory skin disease associated with local infiltration of T lymphocytes producing IL-4 and IL-5, similar to asthma or allergic rhinitis. IL-4 enhances the development of the T helper 2 (Th2) phenotype, resulting in overproduction of immunoglobulins (Ig) and eosinophilia, and increases serum IgE levels.

[0008] Furthermore, since atopic dermatitis is often accompanied by other allergic diseases such as food allergies, asthma, and allergic rhinitis, active control and treatment in childhood is very important. Various treatments are emerging to treat or prevent allergic diseases including atopic dermatitis, but no effective treatment method exists yet. Some drug-based treatments are known, but resistance develops even after short-term use of the medications for treatment, and long-term use can cause significant side effects. Therefore, drug treatment for allergic diseases is recently being avoided. Currently, there is no treatment with absolute and certain efficacy, so short-term treatment is mainly focused on direct irritant symptoms such as itching and redness of the skin other than allergy. Long-term control treatment without side effects is necessary.

[0009] Accordingly, although patent documents 1 and others have disclosed novel strains that exhibit atopic dermatitis treatment effects, strains that exhibit excellent effects in improving allergy symptoms have not yet been discovered, and research to find strains that exhibit the above effects is ongoing in numerous research institutions.

[0010] [Prior Art Literature]

[0011] [Patent Document]

[0012] (Patent Document 0001) Republic of Korea Patent No. 10-2448946

[0013] Against this backdrop, the inventors of the present invention have identified a novel strain with excellent anti-inflammatory activity, anti-allergic activity, and atopic dermatitis treatment effect, and have confirmed the excellent therapeutic effect of the strain, taking note that the health-promoting effect of probiotics is strain-specific rather than a general characteristic of the genus and species, thereby completing the present invention.

[0014] Accordingly, the purpose of the present invention is to provide a Lacticazeibacillus rhamnosus HLL-002 strain deposited under the accession number KCTC15928BP.

[0015] In addition, another object of the present invention is to provide a probiotic composition comprising the aforementioned strain.

[0016] In addition, another object of the present invention is to provide a health functional food composition comprising the aforementioned composition.

[0017] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating allergic diseases comprising the aforementioned strain.

[0018] To solve the above-described problem, the Lacticaseibacillus rhamnosus HLL-002 strain deposited under accession number KCTC15928BP is provided.

[0019] Additionally, the strain may have a 16s rRNA sequence represented by sequence number 1.

[0020] At this time, the strain may have anti-inflammatory or anti-allergic activity.

[0021] At this time, the strain may reduce the secretion of inflammatory cytokines.

[0022] At this time, the inflammatory cytokine may be IL-6 or IL-8.

[0023] At this time, the strain may reduce the level of IgE or ovalbumin-specific IgE.

[0024] Additionally, the present invention provides a probiotic composition comprising at least one selected from the group consisting of the aforementioned strain, strain lysate, strain culture, strain culture concentrate, strain culture extract, and strain dried product.

[0025] At this time, the composition may be anti-inflammatory or anti-allergic.

[0026] Additionally, the present invention provides a health functional food composition comprising the probiotic composition described above.

[0027] Furthermore, the present invention provides a pharmaceutical composition for preventing or treating allergic diseases, comprising at least one selected from the group consisting of the aforementioned strain, strain lysate, strain culture, strain culture concentrate, strain culture extract, and strain dried product.

[0028] At this time, the allergic disease in the composition may be atopic dermatitis.

[0029] At this time, the composition may suppress transdermal moisture loss.

[0030] The novel strain Lacticazeibacillus rhamnosus HLL-002 provided in the present invention or a culture thereof exhibits anti-inflammatory activity by reducing IL-6 and IL-8 levels, reduces transepidermal water loss and clinical scores in the skin, and reduces IgE and ovalbumin-specific IgE in the blood, thereby exhibiting anti-allergic activity and an effect of preventing or treating atopic dermatitis. Therefore, it can be used for anti-inflammatory or anti-allergic purposes, and has the potential to be expanded to a therapeutic agent for preventing or improving allergic diseases, inflammatory diseases, and autoimmune diseases, and thus the novel strain of the present invention has high utility value.

[0031] Figure 1 shows the results of the genome comparison of L. rhamnosusHLL-002 and L. rhamnosusDSM20021 in the form of a pan-genome.

[0032] Figures 2a to 2z and 3a to 3d show different genes in L. rhamnosusHLL-002 and L. rhamnosusDSM20021.

[0033] Figure 4 shows the results of confirming the effect of reducing IL-6 and IL-8 levels in cultures of L. rhamnosusHLL-002 and L. rhamnosusLcr35 strains after inducing inflammation by administering LPS to caco2 cells.

[0034] Figure 5 shows an experimental design diagram for creating an animal model by administering ovalbumin to mice to confirm the atopic dermatitis treatment effect of L. rhamnosusHLL-002.

[0035] Figure 6 shows the results of confirming changes in skin lesions by administration of L. rhamnosus HLL-002 strain and L. rhamnosus GG (LGG) strain in an atopic dermatitis animal model.

[0036] Figure 7 shows the results of confirming changes in transepidermal water loss (TEWL) and skin lesion clinical scores by administration of L. rhamnosus HLL-002 strain and L. rhamnosus GG strain in an atopic dermatitis animal model.

[0037] Figure 8 shows the results of confirming changes in serum total IgE and OVA-specific IgE levels following administration of L. rhamnosus HLL-002 strain and L. rhamnosus GG strain in an atopic dermatitis animal model.

[0038] Hereinafter, the present invention will be described in more detail.

[0039] Meanwhile, each description and embodiment disclosed herein can also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed herein fall within the scope of the present invention. Furthermore, the scope of the present invention is not limited by the specific descriptions described below.

[0040] Furthermore, those skilled in the art will recognize or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments of the invention described in this application. Furthermore, such equivalents are intended to be encompassed by the present invention.

[0041] As described above, the present inventors confirmed that the Lacticaseibacillus rhamnosus HLL-002 strain deposited under accession number KCTC15928BP exhibited a significant atopic dermatitis therapeutic effect in an atopic dermatitis animal model.

[0042] In a specific embodiment of the present invention, the genotypes of Lacticazei Bacillus rhamnosus HLL-002 and Lacticazei Bacillus rhamnosus DSM20021 were compared, and as a result, as shown in FIGS. 1, 2A to 2Z, and 3A to 3D, it was confirmed that it is a new strain with a different genotype from DSM20021.

[0043] Accordingly, the first aspect of the present invention relates to the Lacticaseibacillus rhamnosus HLL-002 strain deposited under accession number KCTC15928BP.

[0044] In the present invention, the strain may have a 16s rRNA sequence represented by sequence number 1.

[0045] In a specific embodiment of the present invention, the anti-inflammatory effect of the Lacticazei Bacillus rhamnosus HLL-002 strain was confirmed. As a result, as shown in Fig. 4, it was confirmed that the HLL-002 strain exhibited an anti-inflammatory effect by reducing the expression levels of IL-6 and IL-8, and it was confirmed that the effect was superior to that of the Lacticazei Bacillus rhamnosus Lcr35 strain, which is currently used as a treatment for atopic dermatitis.

[0046] Therefore, in the present invention, the strain may have anti-inflammatory or anti-allergic activity.

[0047] The term "anti-inflammatory" as used herein means ameliorating (alleviating symptoms), treating, inhibiting or delaying the onset of an inflammatory disease as defined below.

[0048] The term "inflammatory disease" as used herein can be defined as a pathological symptom caused by an inflammatory response that is specific to a local or systemic biological defense response to external physical or chemical stimuli, infection by external infectious agents such as bacteria, fungi, viruses, and various allergens, or autoimmunity. This inflammatory response involves a series of complex physiological reactions such as activation of various inflammatory mediators and enzymes related to immune cells (e.g., iNOS, COX-2, etc.), secretion of inflammatory mediators (e.g., secretion of NO, TNF-α, IL-6, etc.), fluid infiltration, cell migration, tissue destruction, etc., and is externally manifested by symptoms such as erythema, pain, edema, fever, and decline or loss of specific bodily functions. Since the above inflammatory disease may be acute, chronic, ulcerative, allergic, or necrotic, as long as any disease is included in the above definition of inflammatory disease, it does not matter whether it is acute, chronic, ulcerative, allergic, or necrotic. Specifically, the inflammatory diseases may include asthma, allergic and non-allergic rhinitis, chronic and acute rhinitis, chronic and acute gastritis or enteritis, ulcerative gastritis, acute and chronic nephritis, acute and chronic hepatitis, chronic obstructive pulmonary disease, pulmonary fibrosis, irritable bowel syndrome, inflammatory pain, migraine, headache, back pain, fibromyalgia, myofascial disease, viral infection (e.g., hepatitis C infection), bacterial infection, fungal infection, burns, wounds due to surgical or dental work, prostaglandin E hyperactivity syndrome, atherosclerosis, gout, arthritis, rheumatoid arthritis, ankylosing spondylitis, Hodgkin's disease, pancreatitis, conjunctivitis, iritis, scleritis, uveitis, dermatitis (including atopic dermatitis), eczema, multiple sclerosis, etc.

[0049] The term "anti-allergic" as used herein means ameliorating (alleviating symptoms), treating, and preventing (inhibiting or delaying the onset) allergic diseases as defined below.

[0050] The term "allergic disease" as used herein refers to a disease that may occur due to an abnormality in an immune response, and such allergic diseases may include atopic dermatitis, allergic asthma, allergic rhinitis, allergic conjunctivitis, allergic dermatitis, allergic contact dermatitis, food allergy, etc.

[0051] In the present invention, the strain may reduce secretion of inflammatory cytokines.

[0052] In the present invention, the inflammatory cytokine may be IL-6 or IL-8.

[0053] In a specific embodiment of the present invention, as shown in FIG. 5, an atopic dermatitis animal model was created, and the atopic dermatitis therapeutic effect of Lacticazei Bacillus rhamnosus HLL-002 was confirmed. As a result, as shown in FIG. 6, it was confirmed that atopic dermatitis lesions were alleviated by the HLL-002 strain, and it was confirmed through FIG. 7 that TEWL and clinical scores were reduced. In addition, it was confirmed through FIG. 8 that serum total IgE and OVA-specific IgE were reduced by the HLL-002 strain.

[0054] Therefore, in the present invention, the strain may reduce the level of IgE or ovalbumin-specific IgE.

[0055] In addition, the second aspect of the present invention relates to a probiotic composition comprising at least one selected from the group consisting of the strain, strain lysate, strain culture, strain culture concentrate, strain culture extract, and strain dried product.

[0056] Since the effect of the strain included in the probiotic composition of the present invention is the same as that of the monarch of the first aspect, its description is omitted.

[0057] The term "probiotics" used in this invention refers to live microorganisms that, when consumed in sufficient amounts, can have a beneficial effect on health. Most known probiotics are lactic acid bacteria, which, upon reaching the intestines, can perform beneficial functions such as breaking down intestinal food and synthesizing vitamins.

[0058] The probiotic composition of the present invention may be in powder form, and in the case of powder form, it may be coated in the form of a capsule or may not be coated. In addition, the composition may be used in the form of oral administration preparations such as tablets, hard or soft capsules, liquid preparations, suspensions, etc., and these preparations may be prepared using acceptable conventional carriers, for example, in the case of oral administration preparations, excipients, binders, disintegrants, lubricants, solubilizers, suspending agents, preservatives, or bulking agents.

[0059] The term "strain lysate" used in the present invention means a product obtained by culturing a strain and mechanically or chemically disrupting the strain, and may include all products that have undergone additional processes such as extraction, dilution, concentration, and purification therefrom. "Strain culture" may refer to a medium (solid, liquid, etc.) containing a strain, the medium (solid, liquid, etc.) from which the strain is separated after culturing the strain, or the supernatant thereof. "Strain culture concentrate" refers to a product that is purified from a strain culture through ultrafiltration, ammonium sulfate treatment, column purification, concentration, etc., or a culture concentrate obtained through ultrafiltration, concentration, etc. "Strain culture extract" means a product extracted from the culture solution or the concentrate thereof, and may include an extract, a dilution or concentrate of the extract, a dried product obtained by drying the extract, or a preparation or purified product thereof, and a fraction obtained by fractionating the extract. The dried product may include a freeze-dried product.

[0060] In the present invention, the composition may be anti-inflammatory or anti-allergy.

[0061] In addition, the third aspect of the present invention relates to a health functional food composition comprising the probiotic composition.

[0062] The composition and effect of the effective ingredient included in the health functional food composition of the present invention are the same as the composition and effect of the effective ingredient included in the probiotic composition of the second aspect, and therefore, description thereof is omitted.

[0063] The term "food" used in the present invention includes meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, health functional foods, and health foods, and includes all foods in the conventional sense.

[0064] The term "health functional food" used in the present invention means a food manufactured and processed using raw materials or ingredients having functionality useful to the human body according to Act No. 6727 on Health Functional Foods.

[0065] The food composition according to the present invention can be manufactured in various forms according to conventional methods known in the art. General foods include, but are not limited to, beverages (including alcoholic beverages), fruits and processed foods thereof (e.g., canned fruits, bottled fruits, jams, marmalades, etc.), fish, meats and processed foods thereof (e.g., ham, sausages, corned beef, etc.), breads and noodles (e.g., udon, buckwheat noodles, ramen, spaghetti, macaroni, etc.), fruit juices, various drinks, cookies, taffy, dairy products (e.g., butter, cheese, etc.), edible plant oils, margarine, vegetable proteins, retort foods, frozen foods, various seasonings (e.g., soybean paste, soy sauce, sauces, etc.), etc., which can be manufactured by adding the novel strain of the present invention to them. In addition, nutritional supplements include, but are not limited to, capsules, tablets, pills, etc., which can be manufactured by adding the novel strain of the present invention to them. Furthermore, health functional foods are not limited thereto, but for example, the novel strain of the present invention itself can be manufactured into tea, juice, and drinks, and consumed (health drinks) by liquefying, granulating, encapsulating, or powdering. Furthermore, to use the novel strain of the present invention as a food additive, it can be manufactured into a powder or concentrate form.

[0066] When the novel strain of the present invention is used as a health beverage, the health beverage composition may contain various flavoring agents or natural carbohydrates as additional ingredients, similar to conventional beverages. The natural carbohydrates described above may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol, and erythritol. The sweetener may be a natural sweetener such as thaumatin and stevia extract; or a synthetic sweetener such as saccharin and aspartame. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 mL of the composition of the present invention.

[0067] In addition, the novel strain of the present invention may be contained as an effective ingredient of a health functional food composition, and the amount thereof is not particularly limited to an amount effective to achieve an allergic disease prevention or improvement effect, but is preferably 0.01 to 100 wt% based on the total weight of the entire composition. The health functional food composition of the present invention may be prepared by mixing the novel strain with other active ingredients known to be effective against allergic diseases.

[0068] In addition to the above, the health functional food of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonating agents. In addition, the health functional food of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, or vegetable drinks. These ingredients may be used independently or in combination.

[0069] In addition, the fourth aspect of the present invention relates to a pharmaceutical composition for preventing or treating allergic diseases, comprising at least one selected from the group consisting of the strain, strain lysate, strain culture, strain culture concentrate, strain culture extract, and strain dried product.

[0070] The composition and effect of the active ingredient included in the pharmaceutical composition of the present invention are the same as the composition and effect of the active ingredient included in the probiotic composition of the second aspect, and therefore, description thereof is omitted.

[0071] The term 'prevention' used in the present invention means any action that suppresses or delays the onset of an allergic disease or related disease due to the novel strain of the present invention.

[0072] The term 'treatment' used in the present invention means any action that improves or benefits parameters related to an allergy or related disease, such as the degree of symptoms, by the novel strain of the present invention.

[0073] In the present invention, the allergic disease may be an allergic disease mediated by IgE, including bronchial asthma, allergic rhinitis, atopic dermatitis, allergic conjunctivitis, allergic otitis media, urticaria, and anaphylactic shock, or an allergic disease mediated by T cells sensitized to an antigen (or allergen), including contact hypersensitivity, allergic contact dermatitis, bacterial allergy, fungal allergy, viral allergy, drug allergy, thyroiditis, and allergic encephalitis, and preferably an allergic disease mediated by IgE, and most preferably atopic dermatitis.

[0074] The pharmaceutical composition of the present invention may be administered orally or parenterally in various dosage forms. When formulating the composition, it may be prepared using one or more buffers (e.g., saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g., EDTA or glutathione), fillers, bulking agents, binders, adjuvants (e.g., aluminum hydroxide), suspending agents, thickening agents, wetting agents, disintegrating agents, or surfactants, diluents, or excipients.

[0075] Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid dosage forms are prepared by mixing one or more compounds with at least one excipient, such as starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol maltitol, cellulose, methyl cellulose, sodium carboxymethylcellulose, and hydroxypropylmethyl-cellulose or gelatin. For example, tablets or sugar-coated tablets can be obtained by mixing an active ingredient with a solid excipient, grinding the mixture, adding a suitable auxiliary agent, and then processing the mixture into a granule mixture.

[0076] In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, or preservatives may be included. In addition, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as disintegrants in some cases, and anticoagulants, flavoring agents, emulsifiers, solubilizers, dispersants, flavoring agents, antioxidants, packaging agents, pigments, and preservatives may be additionally included.

[0077] Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, or suppositories. Non-aqueous solutions and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include witepsol, macrogol, Tween 61, cocoa butter, laurin, glycerol, and gelatin.

[0078] The composition of the present invention can be administered orally or parenterally, and can be formulated in the form of an injection for parenteral administration into the abdominal cavity, rectum, vein, muscle, or subcutaneously; or a nasal inhaler according to a method known in the art.

[0079] In the case of the above injection, it must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. Examples of suitable carriers for the injection include, but are not limited to, solvents or dispersion media including water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), mixtures thereof, and / or vegetable oils. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine, or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol, and 5% dextrose. In order to protect the injection from microbial contamination, various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, and thimerosal may be additionally included. In addition, the injection may in most cases additionally include isotonic agents such as sugars or sodium chloride.

[0080] For inhalation administration, the compounds used according to the present invention may conveniently be delivered in the form of an aerosol spray from a pressurized pack or nebulizer using a suitable propellant, such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or another suitable gas. For pressurized aerosols, the dosage unit may be determined by providing a valve to deliver a metered amount. For example, gelatin capsules and cartridges for use in inhalers or insufflators may be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch.

[0081] The pharmaceutical composition of the present invention may be a pharmaceutical composition or a quasi-drug composition.

[0082] The term "quasi-drug" used in the present invention refers to products that have a milder effect than pharmaceutical products among products used for the purpose of diagnosing, treating, improving, alleviating, managing or preventing diseases of humans or animals. For example, according to the Pharmaceutical Affairs Act, quasi-drugs are products excluding products used for pharmaceutical purposes, and include products used for treating or preventing diseases of humans or animals, products that have a mild effect on the human body or do not act directly, etc.

[0083] The above-mentioned pharmaceutical composition of the present invention can be manufactured in a formulation selected from the group consisting of body cleanser, disinfectant, detergent, kitchen detergent, cleaning detergent, toothpaste, mouthwash, wet tissue, detergent, soap, hand wash, hair cleanser, hair softener, humidifier filler, mask, ointment, and filter filler, but is not limited thereto.

[0084] The composition of the present invention is administered in a pharmaceutically effective amount. A pharmaceutically effective amount refers to an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment. The effective dosage level can be determined based on the type and severity of the patient's disease, the activity and sensitivity of the drug to the drug, the time of administration, the route of administration and excretion rate, the duration of treatment, concomitant drugs, and other factors well known in the medical field. The composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. That is, the total effective amount of the composition of the present invention can be administered to a patient as a single dose, or can be administered as a fractionated treatment protocol in which multiple doses are administered over a long period of time. It is important to consider all of the above factors and administer an amount that achieves the maximum effect with the minimum amount without causing side effects, and this can be easily determined by those skilled in the art.

[0085] The dosage of the pharmaceutical composition of the present invention may vary depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of the disease.

[0086] In addition, the fifth aspect of the present invention relates to a method for preventing, improving or treating an allergic disease, comprising a step of administering to a subject in need thereof at least one selected from the group consisting of the strain, strain lysate, strain culture, strain culture concentrate, strain culture extract and strain dried product.

[0087] The term "subject" as used herein includes, but is not limited to, any animal (e.g., human, horse, pig, rabbit, dog, sheep, goat, non-human primate, cow, cat, guinea pig, or rodent). This term does not indicate a specific age or gender. Thus, it is intended to include adult / mature and newborn subjects, as well as fetuses, whether female or male.

[0088] In addition, the sixth aspect of the present invention relates to a composition for use in preventing, improving or treating allergic diseases, comprising as an effective ingredient at least one selected from the group consisting of the strain, strain lysate, strain culture, strain culture concentrate, strain culture extract and strain dried product.

[0089] The above composition may be a pharmaceutical composition, a probiotic composition, or a health functional food composition.

[0090] Furthermore, the seventh aspect of the present invention relates to the use of a composition comprising at least one selected from the group consisting of the strain, strain lysate, strain culture, strain culture concentrate, strain culture extract, and strain dried product as an active ingredient for the manufacture of a pharmaceutical, probiotic, or health functional food for preventing, improving, or treating allergic diseases.

[0091] The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

[0092] Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention.

[0093] <Example 1>

[0094] 1. Genotype comparison of L. rhamnosus HLL-002 and L. rhamnosus DSM20021.

[0095] Lactobacillus rhamnosus is a representative intestinal probiotic known to have an intestinal inflammation-relieving effect. L. rhamnosus HLL-002, extracted from a normal Korean child (normal child without a history of allergic disease, family history, or inflammatory disease), has the same 16s rRNA sequence as L. rhamnosus DSM20021, but the whole genome comparative analysis confirmed that it is a new strain with a different genotype from DSM20021. The 16s rRNA sequence of L. rhamnosus HLL-002 is as shown in SEQ ID NO: 1 below, and the results of the genome comparison between the two strains are shown in the pan-genome format in Fig. 1. The presence of a gene is indicated in blue, and the absence is indicated in red, indicating that some genes exist differently in each strain. As shown in Figs. 2a to 2z and 3a to 3d, approximately 700 genes were identified that showed actual differences.

[0096] SEQ ID NO: 1: 16S rRNA (5' CGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATGACTAGTTGCCAGCATTTAGTTGGGCACTCTAGTAAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATCGGATGGTACAACGAGTT GCGAGACCGCGAGGTCAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGACTGTAGGCTGCAACTCGCCTACACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTTGTAACACCCGAAGCCGG 3')

[0097] <Example 2>

[0098] 2. Confirmation of the anti-inflammatory effect of L. rhamnosus HLL-002 strain

[0099] Changes in IL-6 and IL-8 in Caco2 cells by L. rhamnosus HLL-002 strain culture were confirmed using the following method. Caco2 cells were cultured at 1.0x10 in a 60π cell culture dish. 5 The Caco2 cells were cultured in Dulbecco's modified Eagle's minimal essential medium (DMEM) supplemented with 10% FBS (Fetal Bovine Serum) and 1x penicillin-streptomycin for 24 hours. The cultured Caco2 cells were stimulated with LPS, an inflammatory substance, and then treated with L. rhamonsus HLL-002 strain culture and L. rhamnosus Lcr35 strain culture, which is used as a treatment for atopic dermatitis. The cells were then harvested and the gene expression levels of IL-6 and IL-8 were measured using RqPCR.

[0100] As a result, as shown in Fig. 4, when comparing the L. rhamonsusHLL-002 strain culture treatment group with the L. rhamnosusLcr35 strain culture treatment group used as an atopic dermatitis treatment agent, it was confirmed that the anti-inflammatory effect was excellent, confirming its value as a probiotic for controlling inflammation.

[0101] <Example 3>

[0102] 3. Confirmation of the atopic dermatitis treatment effect of L. rhamnosus HLL-002 strain.

[0103] 3-1. Design of an animal model experiment for atopic dermatitis

[0104] To determine the efficacy of the L. rhamonsus HLL-002 strain in atopic dermatitis, an antigen-induced atopic dermatitis animal model was created. Atopic dermatitis was induced through skin sensitization by applying the antigen, ovalbumin (OVA), to the backs of mice. L. rhamonsus HLL-002 was administered orally once daily, starting 14 days before skin sensitization and continuing until the end of the experiment (Fig. 5). L. rhamonsus GG, a commonly used probiotic, served as a positive control.

[0105] 3-2. Confirmation of skin lesion changes due to administration of L. rhamnosus HLL-002 strain in an atopic dermatitis animal model.

[0106] After the completion of antigen sensitization, skin lesions were observed in mice, and it was confirmed that atopic dermatitis lesions were more alleviated in the L. rhamonsusHLL-002 administration group compared to the atopic dermatitis group (AD) and the L. rhamnosusGG group (Fig. 6).

[0107] 3-3. Confirmation of transepidermal water loss and skin lesion clinical scores following administration of L. rhamnosus HLL-002 strain in an atopic dermatitis animal model.

[0108] After antigen sensitization, transepidermal water loss (TEWL), a clinical indicator of atopic dermatitis, and clinical scores within the skin were determined in mice. TEWL was measured using a water evaporation meter, and clinical scores for skin lesions were scored as 0 (no symptoms), 1 (mild), 2 (severe), and 3 (severe) for erythema, redness, and scaling. Compared to the atopic dermatitis (AD) group, the L. rhamnosus HLL-002 administration group showed a decrease in TEWL and clinical scores, and was confirmed to be more effective than the existing L. rhamnosus GG group (Fig. 7).

[0109] 3-4. Confirmation of total serum IgE concentration and OVA-specific IgE concentration by administration of L. rhamnosus HLL-002 strain in an animal model of atopic dermatitis

[0110] To confirm changes in the systemic immune response induced by L. rhamonsusHLL-002 administration, serum total IgE and ovalbumin-specific IgE (OVA-specific IgE) were measured. Total IgE and OVA-specific IgE were measured using the ELISA technique. As shown in Fig. 8, serum total IgE and OVA-specific IgE were confirmed to be reduced in the L. rhamonsusHLL-002-administered group compared to the atopic dermatitis (AD) group.

[0111] The present invention is an invention carried out through the following tasks.

[0112] [National Research and Development Project Supporting This Invention]

[0113] [Project ID] 1711132878

[0114] [Assignment Number] 2017M3A9F3043834

[0115] [Ministry Name] Ministry of Science and ICT

[0116] [Name of Project Management (Specialist) Institution] National Research Foundation of Korea

[0117] [Research Project Name] Bio-Medical Technology Development (R&D)

[0118] [Research Project Name] Discovery of Intestinal Microbiome-Based Pharmabiotics through Immune Mechanism Research in Atopic Dermatitis

[0119] [Name of Project Performing Organization] Ulsan University

[0120] Research Period: August 8, 2017 - May 31, 2022

[0121] [National Research and Development Project Supporting This Invention]

[0122] [Project ID] 1711199653

[0123] [Assignment Number] 2021M3A9I4023974

[0124] [Ministry Name] Ministry of Science and ICT

[0125] [Name of Project Management (Specialist) Institution] National Research Foundation of Korea

[0126] [Research Project Name] Biomedical Technology Development (Support for Research on Next-Generation Promising General-Purpose Biotechnology)

[0127] [Research Project Name] Development of a Microbiome Function Verification Platform Based on a Gut-Mimicking Host Cell-Gut Microbiota Co-Culture System

[0128] [Project Performing Organization Name] Hallym University

[0129] Research Period: April 1, 2021 - December 31, 2025

[0130] [Accession number]

[0131] Name of depositor: Korea Research Institute of Bioscience and Biotechnology, Biological Resource Center (KCTC)

[0132] Accession number: KCTC15928BP

[0133] Date of acceptance: 20240610

[0134]

Claims

1. Lacticaseibacillus rhamnosus HLL-002 strain deposited under accession number KCTC15928BP.

2. A strain according to claim 1, wherein the strain has a 16s rRNA sequence represented by sequence number 1.

3. A strain according to claim 1, wherein the strain has anti-inflammatory or anti-allergic activity.

4. A strain according to claim 1, wherein the strain reduces secretion of inflammatory cytokines.

5. A strain according to claim 4, wherein the inflammatory cytokine is IL-6 or IL-8.

6. A strain according to claim 1, wherein the strain reduces the level of IgE or ovalbumin-specific IgE.

7. A probiotic composition comprising at least one selected from the group consisting of a strain, strain lysate, strain culture, strain culture concentrate, strain culture extract, and strain dried product of any one of claims 1 to 6.

8. A probiotic composition according to claim 7, wherein the composition is for anti-inflammatory or anti-allergy purposes.

9. A health functional food composition comprising the probiotic composition of Article 7.

10. A pharmaceutical composition for preventing or treating allergic diseases, comprising at least one selected from the group consisting of a strain, strain lysate, strain culture, strain culture concentrate, strain culture extract, and strain dried product of any one of claims 1 to 6.

11. A pharmaceutical composition for preventing or treating an allergic disease, wherein the allergic disease is atopic dermatitis in the 10th paragraph.

12. A pharmaceutical composition for preventing or treating allergic diseases, wherein the composition suppresses transepidermal moisture loss in accordance with claim 11.

13. A method for preventing or treating an allergic disease, comprising administering to a subject in need thereof at least one selected from the group consisting of a strain, strain lysate, strain culture, strain culture concentrate, strain culture extract, and strain dried product of any one of claims 1 to 6.

14. A method according to claim 13, wherein the allergic disease is atopic dermatitis.

15. A method according to claim 13, wherein the composition inhibits transepidermal moisture loss.

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