WO2022177411A1 - Pharmaceutical composition for prevention or treatment of cancer comprising weissella cibaria lbml2 strain as active ingredient - Google Patents

Pharmaceutical composition for prevention or treatment of cancer comprising weissella cibaria lbml2 strain as active ingredient Download PDF

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WO2022177411A1
WO2022177411A1 PCT/KR2022/002604 KR2022002604W WO2022177411A1 WO 2022177411 A1 WO2022177411 A1 WO 2022177411A1 KR 2022002604 W KR2022002604 W KR 2022002604W WO 2022177411 A1 WO2022177411 A1 WO 2022177411A1
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cancer
lbml2
strain
composition
weissella cibaria
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PCT/KR2022/002604
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French (fr)
Korean (ko)
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진화섭
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주식회사 리스큐어바이오사이언시스
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Publication of WO2022177411A1 publication Critical patent/WO2022177411A1/en

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

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  • the present invention relates to a composition for preventing or treating cancer comprising a novel Weissella cibaria strain, specifically Weissella cibaria LBML2 (Weissella cibaria LBML2, accession number KCCM12948P) strain or a culture thereof as an active ingredient.
  • a novel Weissella cibaria strain specifically Weissella cibaria LBML2 (Weissella cibaria LBML2, accession number KCCM12948P) strain or a culture thereof as an active ingredient.
  • Cancer has the highest mortality rate worldwide and is the second most common cause of death in Western societies after cardiovascular disease.
  • the consumption of high-fat diets has become common due to the westernization of diets, and the incidence of colorectal cancer, breast cancer, and prostate cancer continues to increase due to the rapid increase in environmental pollutants and the increase in alcohol consumption.
  • Lung cancer is increasing due to an increase in the number of cancer cells and air pollution.
  • lactic acid bacteria are widely distributed in the oral cavity, intestine, vagina, feces, and fermented foods such as kimchi of humans and animals, and are closely related to the health of humans and animals. Lactobacillus has various health-promoting effects such as bowel action, suppression of harmful bacteria, immune regulation, lowering blood cholesterol, and anti-cancer.
  • Korea Patent No. 10-2009742 discloses the anticancer activity of the Weissella civaria strain, but only describes specific examples for the preventive or therapeutic effect on colorectal cancer, and an anticancer therapeutic agent using the novel strain. Or, specific experimental examples or examples have not been described for other carcinomas.
  • Another object of the present invention is to provide a food composition or food additive composition for the prevention or improvement of cancer comprising the Weissella civaria LBML2 strain as an active ingredient.
  • Another object of the present invention is to prevent cancer in livestock containing the Weissella civaria LBML2 strain as an active ingredient.
  • the present inventors have endeavored to find a lactic acid bacteria strain that exhibits cancer prevention and treatment effects, and as a result, a novel Weissella civaria strain having anticancer activity in various carcinomas has been isolated and identified to complete the present invention.
  • the Weissella cibaria LBML2 strain or a pharmaceutical composition for preventing or treating cancer comprising a culture thereof as an active ingredient is provided.
  • the present invention provides a food composition or food additive composition for the prevention or improvement of cancer comprising a Weissella civaria LBML2 strain or a culture thereof as an active ingredient.
  • the present invention provides a feed composition or feed additive composition for preventing or improving cancer of livestock comprising the Weissella civaria LBML2 strain or a culture thereof as an active ingredient.
  • the Weissella cibaria LBML2 strain or a culture thereof according to the present invention exhibits excellent anticancer activity against various carcinomas, it can be usefully used as a composition for use such as treatment, prevention or improvement of cancer in humans or animals. .
  • FIG. 1 is a diagram illustrating a graph measuring a change in the volume of tumor cells over time after intravenous injection of a composition according to an embodiment of the present invention into MC-38 cell transplanted mice.
  • FIG. 2 is a diagram illustrating a graph measuring a change in the volume of tumor cells over time after intravenous injection of a composition according to an embodiment of the present invention into LLC-1 cell-transplanted mice.
  • FIG. 3 is a diagram illustrating a graph measuring the change in the volume of tumor cells over time after intravenous injection of a composition according to an embodiment of the present invention into B16F10 cell-transplanted mice.
  • FIG. 4 is a diagram showing a graph measuring the expression level of the inflammatory cytokine TNF- ⁇ by co-culturing the composition according to an embodiment of the present invention in immature dendritic cells obtained from mice.
  • FIG. 5 is a diagram showing a graph measured using a CCK8 assay whether dendritic cells matured by treatment with the composition according to an embodiment of the present invention can induce T cell division.
  • composition comprising the Weissella cibaria LBML2 strain or a culture thereof of the present invention as an active ingredient has a preventive or therapeutic effect on cancer, and can be used as a pharmaceutical composition.
  • the Weissella civaria LBML2 strain of the present invention is a lactic acid bacteria strain.
  • the Weissella civaria LBML2 strain is a probiotic, and has a general intestinal effect and immune enhancing effect of lactic acid bacteria. It is a well-known fact that the lactic acid bacteria of the Weissella genus have an intestinal effect and immune-enhancing effect.
  • the Wasella civaria strain may be a Wasella civaria LBML2 strain, and as a result of 16S rRNA sequencing for identification and classification of microorganisms through an embodiment of the present invention, SEQ ID NO: 1 of (SEQ ID NO: 1) It has been shown to have a nucleic acid sequence.
  • the microorganism of the present invention having the nucleic acid sequence of SEQ ID NO: 1 was named Weissella civaria LBML2, and was deposited at the Korea Microorganism Conservation Center on February 03, 2021 (Accession No. KCCM12948P).
  • the Wasella civaria LBML2 strain can be used by inoculating 0.1 to 10% in a liquid medium and culturing at 25 to 37° C. for 4 to 48 hours.
  • the culture method is preferably a stationary culture method, but is not limited thereto.
  • 'probiotics' is understood to mean 'living microorganisms that have a beneficial effect on the health of the host by improving the intestinal microbial environment of the host in the gastrointestinal tract of animals including humans'.
  • Probiotics are living microorganisms with probiotic activity, and when fed to humans or animals in the form of single or complex strains, in the form of dried cells or fermentation products, it can have a beneficial effect on the intestinal flora of the host.
  • the cancers include colorectal cancer, lung cancer, black tumor, bladder cancer, breast cancer, thyroid cancer, parathyroid cancer, throat cancer, laryngeal cancer, esophageal cancer, pancreatic cancer, stomach cancer, tongue cancer, skin cancer, brain tumor, uterine cancer, double gallbladder cancer, oral cancer, kidney cancer, liver cancer, colon cancer, It may be any one cancer selected from the group consisting of rectal cancer and blood cancer, but is not limited thereto.
  • the Weissella civaria LBML2 strain included in the composition according to the present invention may exist as a live cell or a dead cell, and may also exist in a dried or lyophilized form.
  • the culture of the Weissella civaria LBML2 strain may be an active ingredient, and the culture may include a live cell culture medium or a dead cell supernatant.
  • Forms of lactic acid bacteria suitable for inclusion in various compositions and methods of formulation are well known to those skilled in the art.
  • composition may be administered orally or parenterally.
  • parenteral administration intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, endothelial administration, topical administration, intranasal administration, intrapulmonary administration and rectal administration, etc. may be administered, preferably by intravenous injection. can be administered, but is not limited thereto.
  • a suitable dosage of the composition may be variously prescribed according to factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity of the patient.
  • the pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant includes an excipient, bog Release, sweetener, binder, coating agent, swelling agent, lubricant, lubricant or flavoring agent and the like can be used.
  • the pharmaceutical composition may be preferably formulated as a pharmaceutical composition by including one or more pharmaceutically acceptable carriers in addition to the active ingredients described above for administration.
  • the active ingredient may be combined with an orally, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like.
  • suitable binders, lubricants, disintegrants and color-developers may also be included in the mixture.
  • suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tracacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate. ate, sodium acetate, sodium chloride, and the like.
  • Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum, and the like.
  • acceptable pharmaceutical carriers which are sterile and biocompatible, include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostats may be added as needed.
  • diluents such as an aqueous solution, suspension, emulsion, etc., pills, capsules, granules or tablets.
  • composition comprising the Weissella cibaria LBML2 strain or a culture thereof of the present invention as an active ingredient may be used as a food composition or food additive composition for preventing or improving cancer.
  • the food composition may be in the form of a health functional food.
  • Health functional food means food manufactured and processed using raw materials or ingredients useful for the human body in accordance with the Health Functional Food Act (Article 3, No. 1), and “functionality” means It refers to obtaining useful effects for health purposes such as regulating nutrients or physiological effects on the structure and function of the human body (Article 2).
  • the food composition may additionally contain food additives, and the suitability as a "food additive" is determined according to the general rules and general test methods of the Food Additives Code approved by the Ministry of Food and Drug Safety, unless otherwise specified. and criteria.
  • Food Additives Codex for example, chemical synthetic products such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as depigmentation, licorice extract, crystalline cellulose, and guar gum, L- Mixed preparations such as sodium glutamate preparation, noodle-added alkali preparation, preservative preparation, and tar color preparation can be mentioned.
  • chemical synthetic products such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid
  • natural additives such as depigmentation, licorice extract, crystalline cellulose, and guar gum
  • L- Mixed preparations such as sodium glutamate preparation, noodle-added alkali preparation, preservative preparation, and tar color preparation can be mentioned.
  • Foods containing the active ingredient of the present invention include bread, rice cakes, dried fruits, candies, chocolates, chewing gum, confectionery products such as jams, ice cream products, ice cream products, ice cream products such as ice cream powder milk, low-fat milk, lactose-decomposed milk, Processed milk, goat milk, fermented milk, buttermilk, concentrated milk, milk cream, butter oil, natural cheese, processed cheese, milk powder, milk products such as whey Processed meat products, processed eggs, meat products such as hamburgers Fish cakes, ham, Fish and meat products such as sausage and bacon processed fish and meat products Ramen, dried noodles, raw noodles, fried noodles, luxurious dried noodles, improved soft noodles, frozen noodles, pasta, etc.
  • the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, It may include a carbonation agent used in carbonated beverages, and the like.
  • the composition of the present invention may include fruit for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination.
  • the beverage composition including the active ingredient of the present invention is not particularly limited in other ingredients, and may contain various flavoring agents or natural carbohydrates as additional ingredients like a conventional beverage.
  • natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.); disaccharides, (eg maltose, sucrose, etc.); and conventional sugars such as polysaccharides (eg, dextrin, cyclodextrin, etc.), and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • natural flavoring agents taumatine, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. .
  • Weissella cibaria LBML2 (Weissella cibaria LBML2) strain of the present invention or a composition comprising a culture thereof as an active ingredient may be used as a feed composition or feed additive composition for preventing or improving cancer in livestock.
  • the composition When the composition is prepared as a feed additive, the composition may be 20 to 90% highly concentrated or may be prepared in powder or granular form.
  • the feed additives include organic acids such as citric acid, humic acid, adipic acid, lactic acid, and malic acid, or phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polyphosphate), polyphenol, catechin, alpha-tocopherol, rosemary Extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, any one or more of natural antioxidants such as phytic acid may be further included.
  • the composition When prepared as a feed, the composition may be formulated in the form of a conventional feed, and may include common feed ingredients together.
  • the feed and feed additives include grains such as milled or crushed wheat, oats, barley, corn and rice; plant protein feeds, such as feeds based on rape, soybean, and sunflower; animal protein feeds such as blood meal, meat meal, bone meal and fish meal; Sugar and dairy products, for example, may further include dry ingredients made of various powdered milk and whey powder, and may further include nutritional supplements, digestion and absorption enhancers, growth promoters, and the like.
  • the feed additive may be administered to the animal alone or in combination with other feed additives in an edible carrier.
  • the feed additive can be easily administered to the animal as a top dressing, directly mixing them with animal feed, or in an oral formulation separate from the feed.
  • a pharmaceutically acceptable edible carrier as well known in the art to prepare an immediate release or sustained release formulation.
  • Such edible carriers may be solid or liquid, for example cornstarch, lactose, sucrose, soybean flakes, peanut oil, olive oil, sesame oil and propylene glycol.
  • the feed additive may be a tablet, capsule, powder, troche or sugar-containing tablet or top dressing in microdispersed form.
  • the feed additive may be in the form of a gelatin soft capsule, or a syrup or suspension, emulsion, or solution.
  • the feed and feed additives may contain adjuvants, for example, preservatives, stabilizers, wetting or emulsifying agents, solution accelerators, and the like.
  • the feed additive may be used by being added to animal feed by immersion, spraying, or mixing.
  • the feed or feed additive of the present invention can be applied to a number of animal diets including mammals, poultry and fish.
  • Pigs, cattle, horses, sheep, rabbits, goats, rodents and experimental rodents as the mammals, as well as rats, hamsters, and guinea pigs can be used for pets (eg, dogs, cats), etc., and as the poultry, chicken, It can also be used for turkey, duck, geese, pheasant, and quail, and the like, and may be used for carp, crucian carp and trout as the fish, but is not limited thereto.
  • the confirmed nucleotide sequence was analyzed for similarity using the Basic Local Alignment Search Tool (BLAST) of the National Center for Biotechnology information (NCBI, www.ncbi.nlm.nih.gov), and through this result, Weissella civaria LBML2 ( Weissella cibaria LBML2) strain was named and deposited at the Korea Microorganism Conservation Center on February 03, 2021.
  • BLAST Basic Local Alignment Search Tool
  • the Weissella cibaria LBML2 (Weissella cibaria LBML2) strain was inoculated with 0.1% in 30ml of a liquid medium and cultured at 37°C for 18 hours. After incubation, centrifugation was performed at 3500 rpm for 10 minutes, and the cells were washed three times with PBS solution and then the remaining medium components were removed.
  • the experimental animals used in the experiment were supplied with 6-week-old male C57bl/6 mice, and were bred during the experiment period after a stabilization period of 1 week in an animal breeding room in an SPF environment with room temperature of 20 ⁇ 2°C and humidity of 55 ⁇ 15%. did.
  • As the feed a normal pellet feed without antibiotics was supplied, and water was provided at any time.
  • Observation of tumor size change was carried out by measuring the volume (mm 3 ) of the tumor using the formula of long axis * short axis 2 /2.
  • MC38 colorectal cancer cells were purchased from Kerafast, and 10% FBS (fetal bovine serum) and 1% antibiotics (penicillin/streptomycin) were added to DMEM medium and cultured at 5% CO 2 , 37°C conditions.
  • FBS fetal bovine serum
  • antibiotics penicillin/streptomycin
  • LLC1 lung cancer cells were purchased from ATCC, and 10% FBS (fetal bovine serum) and 1% antibiotics (penicillin/streptomycin) were added to DMEM medium and cultured at 5% CO 2 , 37°C conditions.
  • FBS fetal bovine serum
  • antibiotics penicillin/streptomycin
  • B16F10 melanoma cells were purchased from ATCC, and 10% FBS (fetal bovine serum) and 1% antibiotics (penicillin/streptomycin) were added to DMEM medium and cultured at 5% CO 2 , 37°C conditions.
  • FBS fetal bovine serum
  • antibiotics penicillin/streptomycin
  • 6-week-old C57bl/6 mice (18-21g) were used in the experiment for the production of the MC-38 cell transplantation animal model.
  • Cultured colon cancer cells MC-38 were harvested (harvest) of 5x10 5 cells, resuspended in 50 ⁇ l of PBS, and injected subcutaneously into the right thigh of the mouse.
  • LLC1 6-week-old C57bl/6 mice (18-21g) were used in the experiment for the production of LLC1 cell transplantation animal models.
  • Cultured colorectal cancer cells LLC1 were harvested (harvest) of 5x10 5 cells, resuspended in 50 ⁇ l PBS, and injected subcutaneously into the right thigh of the mouse.
  • 6-week-old C57bl/6 mice (18-21g) were used in the experiment for the production of the B16F10 melanoma cell transplantation animal model.
  • the cultured colorectal cancer cells B16F10 were harvested (harvest) of 5x10 5 cells, resuspended in 50 ⁇ l of PBS, and injected subcutaneously into the right thigh of the mouse.
  • Weissella cibaria LBML2 (Weissella cibaria LBML2) strain 1x10 9 /100 ul was intravenously injected into the tail of MC-38 colorectal cancer cell-transplanted mice in which the tumor was formed in a volume of about 80-100 mm 3 , and PBS was administered to the negative control group. did. Thereafter, the results of measuring the volume change of the colorectal cancer tumor size of the MC-38 colorectal cancer cell transplanted mice over time are shown in FIG. 1 .
  • Weissella cibaria LBML2 (Weissella cibaria LBML2) strain 1x10 9 /100 ul was intravenously injected into the tail of LLC1 lung cancer cell transplantation mice in which the tumor was formed in a volume of about 80-100 mm 3 , and PBS was administered to the negative control group. Thereafter, the results of measuring the volume change of the lung cancer tumor size of LLC1 lung cancer cell transplantation mice over time are shown in FIG. 2 .
  • Weissella cibaria LBML2 (Weissella cibaria LBML2) strain 1x10 9 /100 ul was intravenously injected into the tail of B16F10 melanoma cell-transplanted mice in which the tumor was formed in a volume of about 80-100 mm 3 , and PBS was administered to the negative control group. Thereafter, the results of measuring the volume change of the melanoma tumor size of the B16F10 melanoma cell transplanted mice over time are shown in FIG. 3 .
  • Example 5 Immature Dendritic Cells maturation Confirmation of activation of induced and differentiated dendritic cells
  • Monocytes were isolated from the bone marrow cells of the mouse femur and tibia by centrifugation using the Ficoll gradient method.
  • the isolated monocytes were prepared in a 6-well plate at a concentration of 2 x 10 6 /well, GM-CSF 20 ng/ml, IL-4 in a culture medium (RPMI) containing Fetal Bovine Serum (FBS) 10 ng/ml were treated together. After treatment, the cells were cultured for 6 days, and when 3 days had elapsed, they were replaced with fresh medium and cytokines to obtain differentiated immature dendritic cells.
  • RPMI culture medium
  • FBS Fetal Bovine Serum
  • the obtained immature dendritic cells (Immature DC) were co-cultured with Weissella cibaria LBML2 strain, a secondary stimulator, for 24 hours for differentiation into mature dendritic cells (mature DC). It was confirmed by measuring the expression of TNFa. The results of measuring the expression of TNFa are shown in FIG. 4 .
  • Example 5-1 To determine whether the dendritic cells matured by the Weissella civaria LBML2 strain in Example 5-1 can induce T cell division, the matured dendritic cells and T cells were co-cultured for 3 days, then CCK8 assay 5 shows the results of measuring the activity and proliferation of T cells using
  • dendritic cells matured by the Weissella civaria LBML2 strain significantly increased the division of T cells, thereby confirming that the anticancer activity would increase by the activated T cells.

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Abstract

The present invention relates to a composition for the prevention or treatment of cancer, comprising a novel Weissella cibaria strain, particularly a Weissella cibaria LBML2 (accession number: KCCM12948P) strain or a culture thereof as an active ingredient. The Weissella cibaria LBML2 strain or the culture thereof according to the present invention exhibits excellent anticancer activity against various carcinomas, and thus can be effectively used as a composition for use in the treatment, prevention, amelioration or the like of cancer in humans or animals.

Description

와이셀라 시바리아 LBML2 균주를 유효성분으로 포함하는 암의 예방 또는 치료용 약학 조성물A pharmaceutical composition for preventing or treating cancer comprising the Wasella civaria LBML2 strain as an active ingredient
본 발명은 신규한 와이셀라 시바리아 균주, 구체적으로 와이셀라 시바리아 LBML2(Weissella cibaria LBML2, 기탁번호 KCCM12948P) 균주 또는 이의 배양물을 유효성분으로 포함하는 암의 예방 또는 치료용 조성물에 관한 것이다.The present invention relates to a composition for preventing or treating cancer comprising a novel Weissella cibaria strain, specifically Weissella cibaria LBML2 (Weissella cibaria LBML2, accession number KCCM12948P) strain or a culture thereof as an active ingredient.
암은 세계적으로 높은 사망률을 보이며, 서구 사회에서는 심혈관 질환 다음으로 가장 일반적인 사망 원인이다. 특히, 식생활이 서구화되어 고지방식의 섭취가 일반화되고, 환경 오염 물질의 급격한 증가, 음주량의 증가 등으로 대장암, 유방암, 전립선암 등이 지속적으로 증가하는 추세에 있으며, 인구의 고령화와 더불어 흡연 인구의 증가 및 대기 오염으로 인해 폐암이 증가하고 있는 실정이다. 이러한 실정에서 암의 조기 예방 및 치료를 가능하게 하여 인간 건강의 증진, 건강한 삶의 질 향상 및 인류 보건 증진에 기여할 수 있는 항암 물질의 창출이 절실히 요구되고 있다.Cancer has the highest mortality rate worldwide and is the second most common cause of death in Western societies after cardiovascular disease. In particular, the consumption of high-fat diets has become common due to the westernization of diets, and the incidence of colorectal cancer, breast cancer, and prostate cancer continues to increase due to the rapid increase in environmental pollutants and the increase in alcohol consumption. Lung cancer is increasing due to an increase in the number of cancer cells and air pollution. In this situation, there is an urgent need for the creation of anticancer substances that can contribute to the promotion of human health, the improvement of healthy life quality, and the promotion of human health by enabling the early prevention and treatment of cancer.
한편, 유산균은 인간과 동물의 구강, 장, 질, 분변, 그리고 김치와 같은 발효식품 등에 널리 분포하면서 사람과 동물의 건강과 밀접한 관련을 맺고 있다. 유산균은 정장작용, 유해균 억제, 면역조절, 혈중 콜레스테롤 저하, 항암 등 다양한 건강증진 효과를 나타내고 있다.On the other hand, lactic acid bacteria are widely distributed in the oral cavity, intestine, vagina, feces, and fermented foods such as kimchi of humans and animals, and are closely related to the health of humans and animals. Lactobacillus has various health-promoting effects such as bowel action, suppression of harmful bacteria, immune regulation, lowering blood cholesterol, and anti-cancer.
현재, 한국등록특허 제10-2009742호에서는 와이셀라 시바리아 균주의 항암 활성을 개시하고 있으나, 대장암에 대한 예방 또는 치료 효과에 대해서만 구체적 실시예를 기재하고 있을 뿐, 신규한 균주를 이용한 항암 치료제 또는 이 외의 암종에 대해서는 구체적인 실험예나 실시예가 기재된 바 없다.Currently, Korea Patent No. 10-2009742 discloses the anticancer activity of the Weissella civaria strain, but only describes specific examples for the preventive or therapeutic effect on colorectal cancer, and an anticancer therapeutic agent using the novel strain. Or, specific experimental examples or examples have not been described for other carcinomas.
본 발명의 목적은 신규한 와이셀라 시바리아 LBML2 균주를 유효성분으로 포함하는 암의 예방 또는 치료용 약학 조성물을 제공하기 위한 것이다.It is an object of the present invention to provide a pharmaceutical composition for preventing or treating cancer comprising a novel Weissella cibaria LBML2 strain as an active ingredient.
또한, 본 발명의 다른 목적은 와이셀라 시바리아 LBML2 균주를 유효성분으로 포함하는 암의 예방 또는 개선용 식품 조성물 또는 식품 첨가제 조성물을 제공하기 위한 것이다.In addition, another object of the present invention is to provide a food composition or food additive composition for the prevention or improvement of cancer comprising the Weissella civaria LBML2 strain as an active ingredient.
또한, 본 발명의 또 다른 목적은 와이셀라 시바리아 LBML2 균주를 유효성분으로 포함하는 가축의 암 예방 또In addition, another object of the present invention is to prevent cancer in livestock containing the Weissella civaria LBML2 strain as an active ingredient.
이에, 본 발명자들은 암의 예방 및 치료 효과를 나타내는 유산균 균주를 찾고자 노력한 결과, 여러 암종에 항암 활성을 갖는 신규한 와이셀라 시바리아 균주를 분리, 동정하여 본 발명을 완성하게 되었다. Accordingly, the present inventors have endeavored to find a lactic acid bacteria strain that exhibits cancer prevention and treatment effects, and as a result, a novel Weissella civaria strain having anticancer activity in various carcinomas has been isolated and identified to complete the present invention.
상기 목적을 달성하기 위하여, 와이셀라 시바리아 LBML2 균주 또는 이의 배양물을 유효성분으로 포함하는 암의 예방 또는 치료용 약학 조성물을 제공한다.In order to achieve the above object, the Weissella cibaria LBML2 strain or a pharmaceutical composition for preventing or treating cancer comprising a culture thereof as an active ingredient is provided.
또한, 본 발명은 와이셀라 시바리아 LBML2 균주 또는 이의 배양물을 유효성분으로 포함하는 암의 예방 또는 개선용 식품 조성물 또는 식품 첨가제 조성물을 제공한다.In addition, the present invention provides a food composition or food additive composition for the prevention or improvement of cancer comprising a Weissella civaria LBML2 strain or a culture thereof as an active ingredient.
또한, 본 발명은 와이셀라 시바리아 LBML2 균주 또는 이의 배양물을 유효성분으로 포함하는 가축의 암 예방 또는 개선용 사료 조성물 또는 사료 첨가제 조성물을 제공한다.In addition, the present invention provides a feed composition or feed additive composition for preventing or improving cancer of livestock comprising the Weissella civaria LBML2 strain or a culture thereof as an active ingredient.
본 발명에 따른 와이셀라 시바리아 LBML2 균주 또는 이의 배양물은 여러 암종에 대해 우수한 항암 활성을 나타내므로, 사람 또는 동물의 암의 치료, 예방 또는 개선 등의 용도를 위한 조성물로서 유용하게 이용될 수 있다.Since the Weissella cibaria LBML2 strain or a culture thereof according to the present invention exhibits excellent anticancer activity against various carcinomas, it can be usefully used as a composition for use such as treatment, prevention or improvement of cancer in humans or animals. .
도 1은 본 발명의 일 실시예에 따른 조성물을 MC-38 세포 이식 마우스에 정맥 주사한 후 시간이 경과함에 따라 종양 세포의 부피 변화를 측정한 그래프를 나타낸 도이다.1 is a diagram illustrating a graph measuring a change in the volume of tumor cells over time after intravenous injection of a composition according to an embodiment of the present invention into MC-38 cell transplanted mice.
도 2는 본 발명의 일 실시예에 따른 조성물을 LLC-1 세포 이식 마우스에 정맥 주사한 후 시간이 경과함에 따라 종양 세포의 부피 변화를 측정한 그래프를 나타낸 도이다.2 is a diagram illustrating a graph measuring a change in the volume of tumor cells over time after intravenous injection of a composition according to an embodiment of the present invention into LLC-1 cell-transplanted mice.
도 3은 본 발명의 일 실시예에 따른 조성물을 B16F10 세포 이식 마우스에 정맥 주사한 후 시간이 경과함에 따라 종양 세포의 부피 변화를 측정한 그래프를 나타낸 도이다.3 is a diagram illustrating a graph measuring the change in the volume of tumor cells over time after intravenous injection of a composition according to an embodiment of the present invention into B16F10 cell-transplanted mice.
도 4는 본 발명의 일 실시예에 따른 조성물을 마우스로부터 수득한 미성숙 수지상 세포에 공동배양하여 염증성 사이토카인 TNF-α 발현 정도를 측정한 그래프를 나타낸 도이다.4 is a diagram showing a graph measuring the expression level of the inflammatory cytokine TNF-α by co-culturing the composition according to an embodiment of the present invention in immature dendritic cells obtained from mice.
도 5는 본 발명의 일 실시예에 따른 조성물을 처리하여 성숙화된 수지상 세포가 T 세포의 분열을 유도할 수 있는지 CCK8 assay를 이용하여 측정한 그래프를 나타낸 도이다.5 is a diagram showing a graph measured using a CCK8 assay whether dendritic cells matured by treatment with the composition according to an embodiment of the present invention can induce T cell division.
본 발명의 와이셀라 시바리아 LBML2(Weissella cibaria LBML2) 균주 또는 이의 배양물을 유효성분으로 포함하는 조성물은 암의 예방 또는 치료 효과를 가지며, 약제학적 조성물로 이용될 수 있다.The composition comprising the Weissella cibaria LBML2 strain or a culture thereof of the present invention as an active ingredient has a preventive or therapeutic effect on cancer, and can be used as a pharmaceutical composition.
본 발명의 와이셀라 시바리아 LBML2 균주는 유산균 균주이다. 상기 와이셀라 시바리아 LBML2 균주는 프로바이오틱스로서, 유산균의 일반적인 정장 효과 및 면역 증강 효과를 갖는다. 와이셀라 속의 유산균이 정장 효과 면역 증강 효과를 갖는 것은 잘 알려진 사실이다.The Weissella civaria LBML2 strain of the present invention is a lactic acid bacteria strain. The Weissella civaria LBML2 strain is a probiotic, and has a general intestinal effect and immune enhancing effect of lactic acid bacteria. It is a well-known fact that the lactic acid bacteria of the Weissella genus have an intestinal effect and immune-enhancing effect.
상기 와이셀라 시바리아 균주는 와이셀라 시바리아 LBML2 균주일 수 있으며, 본 발명의 실시예를 통해 미생물의 동정 및 분류를 위한 16S rRNA 염기서열 분석 결과, 서열번호 1의(SEQ ID NO: 1)의 핵산서열을 갖는 것으로 나타났다.The Wasella civaria strain may be a Wasella civaria LBML2 strain, and as a result of 16S rRNA sequencing for identification and classification of microorganisms through an embodiment of the present invention, SEQ ID NO: 1 of (SEQ ID NO: 1) It has been shown to have a nucleic acid sequence.
상기 서열번호 1의 핵산서열을 갖는 본 발명의 미생물을 와이셀라 시바리아 LBML2로 명명하였으며, 한국미생물보존센터에 2021년 02월 03일자로 기탁하였다(기탁번호 KCCM12948P).The microorganism of the present invention having the nucleic acid sequence of SEQ ID NO: 1 was named Weissella civaria LBML2, and was deposited at the Korea Microorganism Conservation Center on February 03, 2021 (Accession No. KCCM12948P).
상기 와이셀라 시바리아 LBML2 균주는 액체배지에 0.1 내지 10% 접종하여 25 내지 37℃에서 4시간 내지 48시간 동안 배양하여 이용할 수 있다. The Wasella civaria LBML2 strain can be used by inoculating 0.1 to 10% in a liquid medium and culturing at 25 to 37° C. for 4 to 48 hours.
상기 배양의 방법은 정치배양 방법이 바람직하나, 이에 한정되지 않는다. The culture method is preferably a stationary culture method, but is not limited thereto.
본 발명에 있어서, '프로바이오틱스(probiotics)'는 사람을 포함한 동물의 위장관 내에서 숙주의 장내 미생물 환경을 개선하여 숙주의 건강에 유익한 영향을 주는 살아있는 미생물'이라는 의미로 이해된다. 프로바이오틱스는 프로바이오틱 활성을 갖는 살아있는 미생물로 단일 또는 복합균주 형태로 사람이나 동물에 건조된 세포 형태나 발효산물 형태로 급여될 경우, 숙주의 장내 균총에 유익한 영향을 미칠 수 있다.In the present invention, 'probiotics' is understood to mean 'living microorganisms that have a beneficial effect on the health of the host by improving the intestinal microbial environment of the host in the gastrointestinal tract of animals including humans'. Probiotics are living microorganisms with probiotic activity, and when fed to humans or animals in the form of single or complex strains, in the form of dried cells or fermentation products, it can have a beneficial effect on the intestinal flora of the host.
상기 암은 대장암, 폐암, 흑색종양, 방광암, 유방암, 갑상선암, 부갑상선암, 인후암, 후두암, 식도암, 췌장암, 위암, 설암, 피부암, 뇌종양, 자궁암, 두담낭암, 구강암, 신장암, 간암, 결장암, 직장암, 및 혈액암으로 이루어지는 군으로부터 선택되는 어느 하나의 암일 수 있으며, 이에 한정되지 않는다.The cancers include colorectal cancer, lung cancer, black tumor, bladder cancer, breast cancer, thyroid cancer, parathyroid cancer, throat cancer, laryngeal cancer, esophageal cancer, pancreatic cancer, stomach cancer, tongue cancer, skin cancer, brain tumor, uterine cancer, double gallbladder cancer, oral cancer, kidney cancer, liver cancer, colon cancer, It may be any one cancer selected from the group consisting of rectal cancer and blood cancer, but is not limited thereto.
본 발명에 따른 조성물에 포함되는 와이셀라 시바리아 LBML2 균주는 생균체 또는 사균체로서 존재할 수 있으며, 또한 건조 또는 동결건조된 형태로 존재할 수도 있다. 또한, 와이셀라 시바리아 LBML2 균주의 배양물이 유효성분일 수 있으며, 상기 배양물에는 생균 배양액 또는 사균 상등액이 포함될 수 있다. 다양한 조성물 내에 포함시키기 적합한 유산균의 형태 및 제제화 방법은 당업자에게 잘 알려져 있다.The Weissella civaria LBML2 strain included in the composition according to the present invention may exist as a live cell or a dead cell, and may also exist in a dried or lyophilized form. In addition, the culture of the Weissella civaria LBML2 strain may be an active ingredient, and the culture may include a live cell culture medium or a dead cell supernatant. Forms of lactic acid bacteria suitable for inclusion in various compositions and methods of formulation are well known to those skilled in the art.
상기 조성물은 경구 또는 비경구로 투여할 수 있다. 비경구 투여인 경우에는 정맥 내 주입, 피하 주입, 근육 주입, 복강 주입, 내피 투여, 국소 투여, 비내 투여, 폐 내 투여 및 직장 내 투여 등으로 투여할 수 있으며, 바람직하게는 정맥 내 주입방법으로 투여할 수 있으나, 이에 한정되지 않는다.The composition may be administered orally or parenterally. In the case of parenteral administration, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, endothelial administration, topical administration, intranasal administration, intrapulmonary administration and rectal administration, etc. may be administered, preferably by intravenous injection. can be administered, but is not limited thereto.
상기 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여시간, 투여경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다.A suitable dosage of the composition may be variously prescribed according to factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity of the patient.
본 발명의 조성물이 약제학적 조성물로 활용될 경우, 본 발명의 약제학적 조성물은 상기 유효성분 이외에 약제학적으로 적합하고 생리학적으로 허용되는 보조제를 사용하여 제조될 수 있으며, 상기 보조제로는 부형제, 붕해제, 감미제, 결합제, 피복제, 팽창제, 윤활제, 활택제 또는 향미제 등을 사용할 수 있다.When the composition of the present invention is utilized as a pharmaceutical composition, the pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant includes an excipient, bog Release, sweetener, binder, coating agent, swelling agent, lubricant, lubricant or flavoring agent and the like can be used.
상기 약제학적 조성물은 투여를 위해서 상기 기재한 유효성분 이외에 추가로 약제학적으로 허용가능한 담체를 1종 이상 포함하여 약제학적 조성물로 바람직하게 제제화할 수 있다.The pharmaceutical composition may be preferably formulated as a pharmaceutical composition by including one or more pharmaceutically acceptable carriers in addition to the active ingredients described above for administration.
예를 들어, 정제 또는 캡슐제의 형태로의 제제화를 위해, 유효성분은 에탄올, 글리세롤, 물 등과 같은 경구, 무독성의 약제학적으로 허용가능한 불활성 담체와 결합될 수 있다. 또한, 원하거나 필요한 경우, 적합한 결합제, 윤활제, 붕해제 및 발색제 또한 혼합물로 포함될 수 있다. 적합한 결합제는 이에 제한되는 것은 아니나, 녹말, 젤라틴, 글루코스 또는 베타-락토오스와 같은 천연당, 옥수수감미제, 아카시아, 트래커캔스 또는 소듐올레이트와 같은 천연 및 합성검, 소듐 스테아레이트, 마그네슘 스테아레이트, 소듐 벤조에이트, 소듐아세테이트, 소듐클로라이드 등을 포함한다. 붕해제는 이에 제한되는 것은 아니나, 녹말, 메틸셀룰로스, 아가, 벤토니트, 잔탄검 등을 포함한다. 액상 용액으로 제제화되는 조성물에 있어서 허용가능한 약제학적 담체로는, 멸균 및 생체에 적합한 것으로서, 식염수, 멸균수, 링거액, 완충식염수, 알부민 주사 용액, 덱스트로즈 용액, 말토덱스트린 용액, 글리세롤, 에탄올 및 이들 성분 중 1 성분이상을 혼합하여 사용할 수 있으며, 필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가할 수 있다. 또한 희석제, 분산제, 계면활성제, 결합제 및 윤활제를 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화할 수 있다.For example, for formulation in the form of a tablet or capsule, the active ingredient may be combined with an orally, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or required, suitable binders, lubricants, disintegrants and color-developers may also be included in the mixture. Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tracacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate. ate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum, and the like. In the composition formulated as a liquid solution, acceptable pharmaceutical carriers, which are sterile and biocompatible, include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostats may be added as needed. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to form an injectable formulation such as an aqueous solution, suspension, emulsion, etc., pills, capsules, granules or tablets.
나아가 해당 분야의 적절한 방법으로 Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA에 개시되어 있는 방법을 이용하여 각 질환에 따라 또는 성분에 따라 바람직하게 제제화할 수 있다.Furthermore, it can be preferably formulated according to each disease or component using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA by an appropriate method in the field.
본 발명의 와이셀라 시바리아 LBML2(Weissella cibaria LBML2) 균주 또는 이의 배양물을 유효성분으로 포함하는 조성물은 암의 예방 또는 개선용 식품 조성물 또는 식품 첨가제 조성물로 이용될 수 있다.The composition comprising the Weissella cibaria LBML2 strain or a culture thereof of the present invention as an active ingredient may be used as a food composition or food additive composition for preventing or improving cancer.
상기 식품 조성물은 건강기능식품의 형태일 수 있다.The food composition may be in the form of a health functional food.
상기 "건강기능식품"은 건강기능식품에 관한 법률에 따라 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미(제3조 제1호)하며, "기능성"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻는 것(동조 제2호)을 의미한다.The above "health functional food" means food manufactured and processed using raw materials or ingredients useful for the human body in accordance with the Health Functional Food Act (Article 3, No. 1), and "functionality" means It refers to obtaining useful effects for health purposes such as regulating nutrients or physiological effects on the structure and function of the human body (Article 2).
상기 식품 조성물은 식품 첨가물을 추가로 포함할 수 있으며, "식품첨가물"로서의 적합여부는 다른 규정이 없는 한 식품의약품안전처에 승인된 식품첨가물공전의 총칙 및 일반시험법 등에 따라 해당 품목에 관한 규격 및 기준에 의하여 판정한다.The food composition may additionally contain food additives, and the suitability as a "food additive" is determined according to the general rules and general test methods of the Food Additives Code approved by the Ministry of Food and Drug Safety, unless otherwise specified. and criteria.
상기 "식품첨가물공전"에 수재된 품목으로 예를 들어, 케톤류, 글리신, 구연산칼륨, 니코틴산, 계피산 등의 화학적 합성품, 감색소, 감초추출물, 결정셀롤로오스, 구아검 등의 천연첨가물, L-글루타민산나트륨제제, 면류첨가알칼리제, 보존료제제, 타르색소제제 등의 혼합 제제류 들을 들 수 있다.As items listed in the "Food Additives Codex", for example, chemical synthetic products such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as depigmentation, licorice extract, crystalline cellulose, and guar gum, L- Mixed preparations such as sodium glutamate preparation, noodle-added alkali preparation, preservative preparation, and tar color preparation can be mentioned.
본 발명의 유효성분이 포함된 식품으로는 빵, 떡류, 건과류, 캔디류, 초콜릿류, 츄잉껌, 쨈류와 같은 과자류 아이스크림류, 빙과류, 아이스크림 분말류와 같은 아이스크림 제품류 우유류, 저지방 우유류, 유당분해 우유, 가공유류, 산양유, 발효유류, 버터유류, 농축유류, 유크림류, 버터유, 자연치즈, 가공치즈, 분유류, 유청류와 같은 유가공품류 식육가공품, 알가공품, 햄버거와 같은 식육제품류 어묵, 햄, 소세지, 베이컨 등의 어육가공품과 같은 어육제품류 라면류, 건면류, 생면류, 유탕면류, 호화건먼류, 개량숙면류, 냉동면류, 파스타류와 같은 면류 과실음료, 채소류음료, 탄산음료, 두유류, 요구르트 등의 유산균음료, 혼합음료와 같은 음료 간장, 된장, 고추장, 춘장, 청국장, 혼합장, 식초, 소스류, 토마토케첩, 카레, 드레싱과 같은 조미식품 마가린, 쇼트닝 및 피자를 들 수 있으나, 이에 한정되는 것은 아니다.Foods containing the active ingredient of the present invention include bread, rice cakes, dried fruits, candies, chocolates, chewing gum, confectionery products such as jams, ice cream products, ice cream products, ice cream products such as ice cream powder milk, low-fat milk, lactose-decomposed milk, Processed milk, goat milk, fermented milk, buttermilk, concentrated milk, milk cream, butter oil, natural cheese, processed cheese, milk powder, milk products such as whey Processed meat products, processed eggs, meat products such as hamburgers Fish cakes, ham, Fish and meat products such as sausage and bacon processed fish and meat products Ramen, dried noodles, raw noodles, fried noodles, luxurious dried noodles, improved soft noodles, frozen noodles, pasta, etc. Fruit drinks, vegetable drinks, carbonated drinks, soy milk, Lactic acid bacteria drinks such as yogurt, beverages such as mixed drinks Soy sauce, soybean paste, gochujang, chunjang, cheonggukjang, mixed soy sauce, vinegar, sauces, tomato ketchup, curry, seasoning foods such as dressings, margarine, shortening and pizza, but limited to this it's not going to be
상기 외에 본 발명의 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 포함할 수 있다. 그 밖에 본 발명의 조성물은 천연 과일주스, 과일주스 음료 및 야채 음료의 제조를 위한 과육을 포함할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, It may include a carbonation agent used in carbonated beverages, and the like. In addition, the composition of the present invention may include fruit for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination.
본 발명의 유효성분을 포함한 음료 조성물은 다른 성분에는 특별한 제한이 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가성분으로 함유할 수 있다. 상술한 천연 탄수화물의 예는 모노사카라이드, (예를 들어, 포도당, 과당 등); 디사카라이드, (예를 들어 말토스, 슈크로스 등); 및 폴리사카라이드, (예를 들어 덱스트린, 시클로덱스트린 등)과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 것 이외의 향미제로서 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진 등) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다.The beverage composition including the active ingredient of the present invention is not particularly limited in other ingredients, and may contain various flavoring agents or natural carbohydrates as additional ingredients like a conventional beverage. Examples of the aforementioned natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.); disaccharides, (eg maltose, sucrose, etc.); and conventional sugars such as polysaccharides (eg, dextrin, cyclodextrin, etc.), and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatine, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. .
또한, 본 발명의 와이셀라 시바리아 LBML2(Weissella cibaria LBML2) 균주 또는 이의 배양물을 유효성분으로 포함하는 조성물은 가축의 암 예방 또는 개선용 사료 조성물 또는 사료 첨가제 조성물로 이용될 수 있다.In addition, the Weissella cibaria LBML2 (Weissella cibaria LBML2) strain of the present invention or a composition comprising a culture thereof as an active ingredient may be used as a feed composition or feed additive composition for preventing or improving cancer in livestock.
상기 조성물이 사료 첨가제로서 제조될 경우, 상기 조성물은 20 내지 90% 고농축액이거나 분말 또는 과립 형태로 제조될 수 있다. 상기 사료 첨가제는 구연산, 후말산, 아디픽산, 젖산, 사과산등의 유기산이나 인산 나트륨, 인산 칼륨, 산성 피로인산염, 폴리인산염(중합인산염) 등의 인산염이나, 폴리페놀, 카테킨, 알파-토코페롤, 로즈마리 추출물, 비타민 C, 녹차 추출물, 감초 추출물, 키토산, 탄닌산, 피틴산 등의 천연 항산화제 중 어느 하나 또는 하나 이상을 추가로 포함할 수 있다. 사료로서 제조될 경우, 상기 조성물은 통상의 사료 형태로 제제화될 수 있으며, 통상의 사료 성분을 함께 포함할 수 있다.When the composition is prepared as a feed additive, the composition may be 20 to 90% highly concentrated or may be prepared in powder or granular form. The feed additives include organic acids such as citric acid, humic acid, adipic acid, lactic acid, and malic acid, or phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polyphosphate), polyphenol, catechin, alpha-tocopherol, rosemary Extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, any one or more of natural antioxidants such as phytic acid may be further included. When prepared as a feed, the composition may be formulated in the form of a conventional feed, and may include common feed ingredients together.
상기 사료 및 사료 첨가제는 곡물, 예를 들면 분쇄 또는 파쇄된 밀, 귀리, 보리, 옥수수 및 쌀; 식물성 단백질 사료, 예를 들면 평지, 콩, 및 해바라기를 주성분으로 하는 사료; 동물성 단백질 사료, 예를 들면 혈분, 육분, 골분 및 생선분; 당분 및 유제품, 예를 들면 각종 분유 및 유장 분말로 이루어지는 건조성분 등을 더 포함할 수 있으며, 이외에도 영양보충제, 소화 및 흡수향상제, 성장촉진제 등을 더 포함할 수 있다.The feed and feed additives include grains such as milled or crushed wheat, oats, barley, corn and rice; plant protein feeds, such as feeds based on rape, soybean, and sunflower; animal protein feeds such as blood meal, meat meal, bone meal and fish meal; Sugar and dairy products, for example, may further include dry ingredients made of various powdered milk and whey powder, and may further include nutritional supplements, digestion and absorption enhancers, growth promoters, and the like.
상기 사료 첨가제는 동물에게 단독으로 투여하거나 식용 담체 중에서 다른 사료 첨가제와 조합하여 투여할 수도 있다. 또한, 상기 사료 첨가제는 탑드레싱으로서 또는 이들을 동물사료에 직접 혼합하거나 또는 사료와 별도의 경구제형으로 용이하게 동물에게 투여할 수 있다. 상기 사료 첨가제를 동물사료와 별도로 투여할 경우, 당해 기술분야에 잘 알려진 바와 같이 약제학적으로 허용 가능한 식용 담체와 조합하여, 즉시 방출 또는 서방성 제형으로 제조할 수 있다. 이러한 식용 담체는 고체 또는 액체, 예를 들어 옥수수전분, 락토오스, 수크로오스, 콩플레이크, 땅콩유, 올리브유, 참깨유 및 프로필렌글리콜일 수 있다. 고체 담체가 사용될 경우, 사료 첨가제는 정제, 캡슐제, 산제, 트로키제 또는 함당정제 또는 미분산성 형태의 탑 드레싱일 수 있다. 액체 담체가 사용될 경우, 사료 첨가제는 젤라틴 연질 캡슐제, 또는 시럽제나 현탁액, 에멀젼제, 또는 용액제의 제형일 수 있다.The feed additive may be administered to the animal alone or in combination with other feed additives in an edible carrier. In addition, the feed additive can be easily administered to the animal as a top dressing, directly mixing them with animal feed, or in an oral formulation separate from the feed. When the feed additive is administered separately from animal feed, it can be combined with a pharmaceutically acceptable edible carrier as well known in the art to prepare an immediate release or sustained release formulation. Such edible carriers may be solid or liquid, for example cornstarch, lactose, sucrose, soybean flakes, peanut oil, olive oil, sesame oil and propylene glycol. When a solid carrier is used, the feed additive may be a tablet, capsule, powder, troche or sugar-containing tablet or top dressing in microdispersed form. When a liquid carrier is used, the feed additive may be in the form of a gelatin soft capsule, or a syrup or suspension, emulsion, or solution.
또한, 상기 사료 및 사료 첨가제는 보조제, 예를 들어 보존제, 안정화제, 습윤제 또는 유화제, 용액 촉진제 등을 함유할 수 있다. 상기 사료 첨가제는 침주, 분무 또는 혼합하여 동물의 사료에 첨가하여 이용될 수 있다.In addition, the feed and feed additives may contain adjuvants, for example, preservatives, stabilizers, wetting or emulsifying agents, solution accelerators, and the like. The feed additive may be used by being added to animal feed by immersion, spraying, or mixing.
본 발명의 사료 또는 사료 첨가제는 포유류, 가금 및 어류를 포함하는 다수의 동물 식이에 적용할 수 있다.The feed or feed additive of the present invention can be applied to a number of animal diets including mammals, poultry and fish.
상기 포유류로서 돼지, 소, 말, 양, 토끼, 염소, 설치동물 및 실험용 설치동물인 쥐, 햄스터, 기니피그 뿐만 아니라, 애완동물(예: 개, 고양이) 등에게 사용할 수 있으며, 상기 가금류로서 닭, 칠면조, 오리, 거위, 꿩, 및 메추라기 등에도 사용할 수 있고, 상기 어류로서 잉어, 붕어 및 송어 등에 이용될 수 있으나, 이에 한정되는 것은 아니다.Pigs, cattle, horses, sheep, rabbits, goats, rodents and experimental rodents as the mammals, as well as rats, hamsters, and guinea pigs, can be used for pets (eg, dogs, cats), etc., and as the poultry, chicken, It can also be used for turkey, duck, geese, pheasant, and quail, and the like, and may be used for carp, crucian carp and trout as the fish, but is not limited thereto.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로서, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .
실시예Example 1. 균주의 분리 및 동정 1. Isolation and identification of strains
여러 종류의 김치 시료를 파쇄한 후, 김치액을 PBS(phosphate buffered saline)로 연속희석(serial dilution, 103~108)하여 1차 젖산균 선택 배지에 도말한 후 30℃ 또는 37℃에서 각각 24시간 배양하였다. 형성된 콜로니를 단일 콜로니로 분리한 뒤, 류코노스톡 선택 배지인 Phenylethyl alcohol sucrose agar 배지에 선상 도말하여 20℃에서 24시간 배양하였다. 배양 후, 형성된 단일 콜로니를 수거하여 액체배지에 넣고 배양하였다. 24시간 배양한 배양액으로부터 genomic DNA를 추출하였다. 16S rDNA 염기서열을 확인하기 위해, 얻어진 genomic DNA 산물은 마크로젠에 분석 의뢰하였다. 확인한 염기서열을 National Center for Biotechnology information (NCBI, www.ncbi.nlm.nih.gov)의 Basic Local Alignment Search Tool (BLAST)을 이용하여 유사도 분석하였고, 이 결과를 통해 최종적으로 와이셀라 시바리아 LBML2(Weissella cibaria LBML2) 균주로 명명하고, 한국미생물 보존센터에 2021년 2월 03일자로 기탁하였다.After crushing various types of kimchi samples, serial dilution (103~108) of the kimchi solution with PBS (phosphate buffered saline) is applied to the primary lactic acid bacteria selection medium, and then cultured at 30°C or 37°C for 24 hours, respectively. did. After the formed colonies were separated into single colonies, they were streaked on a Phenylethyl alcohol sucrose agar medium, which is a leukonostok selective medium, and cultured at 20° C. for 24 hours. After culturing, the formed single colonies were collected and cultured in a liquid medium. Genomic DNA was extracted from the culture medium cultured for 24 hours. To confirm the 16S rDNA base sequence, the obtained genomic DNA product was requested for analysis by Macrogen. The confirmed nucleotide sequence was analyzed for similarity using the Basic Local Alignment Search Tool (BLAST) of the National Center for Biotechnology information (NCBI, www.ncbi.nlm.nih.gov), and through this result, Weissella civaria LBML2 ( Weissella cibaria LBML2) strain was named and deposited at the Korea Microorganism Conservation Center on February 03, 2021.
실시예Example 2. 미생물 균주의 배양 2. Cultivation of microbial strains
상기 와이셀라 시바리아 LBML2 (Weissella cibaria LBML2) 균주를 액체배지 30ml에 0.1% 접종하여 37℃에서 18시간 동안 배양하였다. 배양 후, 3500 rpm에서 10분 동안 원심분리하고, 균체는 PBS 용액으로 3회 세척한 후 남아 있는 배지 성분을 제거하여 수득하였다.The Weissella cibaria LBML2 (Weissella cibaria LBML2) strain was inoculated with 0.1% in 30ml of a liquid medium and cultured at 37°C for 18 hours. After incubation, centrifugation was performed at 3500 rpm for 10 minutes, and the cells were washed three times with PBS solution and then the remaining medium components were removed.
실시예Example 3. 실험동물 (마우스)의 조건 3. Conditions of experimental animals (mouse)
실험에 사용된 실험동물은 수컷 6주령의 C57bl/6 마우스를 공급받았고, 실내 온도 20±2℃ 습도 55±15%가 유지되는 SPF 환경의 동물 사육실에서 1주일의 안정화 기간을 거쳐 실험기간 동안 사육하였다. 사료는 항생제가 첨가되지 않은 일반적인 팰렛 사료를 공급하였고, 물은 수시로 섭취할 수 있게 하였다. 종양 크기 변화의 관찰은 장축*단축2/2의 식을 사용하여 종양의 부피(mm3)를 측정하여 진행하였다.The experimental animals used in the experiment were supplied with 6-week-old male C57bl/6 mice, and were bred during the experiment period after a stabilization period of 1 week in an animal breeding room in an SPF environment with room temperature of 20±2℃ and humidity of 55±15%. did. As the feed, a normal pellet feed without antibiotics was supplied, and water was provided at any time. Observation of tumor size change was carried out by measuring the volume (mm 3 ) of the tumor using the formula of long axis * short axis 2 /2.
실시예Example 4. 4. 암 세포cancer cells 이식 마우스 모델에서의 항종양 효과 분석 Analysis of antitumor effect in transplanted mouse model
4-1. 세포배양4-1. cell culture
MC38 대장암 세포는 Kerafast에서 구입하여 사용하였으며, DMEM 배지에 10% FBS(fetal bovine serum)와 1% 항생제(penicillin/streptomycin)를 첨가하여 5% CO2, 37℃ 조건에서 배양하였다.MC38 colorectal cancer cells were purchased from Kerafast, and 10% FBS (fetal bovine serum) and 1% antibiotics (penicillin/streptomycin) were added to DMEM medium and cultured at 5% CO 2 , 37°C conditions.
LLC1 폐암 세포는 ATCC에서 구입하여 사용하였으며, DMEM 배지에 10% FBS(fetal bovine serum)와 1% 항생제(penicillin/streptomycin)를 첨가하여 5% CO2, 37℃ 조건에서 배양하였다.LLC1 lung cancer cells were purchased from ATCC, and 10% FBS (fetal bovine serum) and 1% antibiotics (penicillin/streptomycin) were added to DMEM medium and cultured at 5% CO 2 , 37°C conditions.
B16F10 흑색종 세포는 ATCC에서 구입하여 사용하였으며, DMEM 배지에 10% FBS(fetal bovine serum)와 1% 항생제(penicillin/streptomycin)를 첨가하여 5% CO2, 37℃ 조건에서 배양하였다.B16F10 melanoma cells were purchased from ATCC, and 10% FBS (fetal bovine serum) and 1% antibiotics (penicillin/streptomycin) were added to DMEM medium and cultured at 5% CO 2 , 37°C conditions.
4-2. 4-2. 암 세포cancer cells 이식 마우스 모델 제작 Construction of an implanted mouse model
MC-38 세포 이식 동물모델 제작을 위해서 6주령 C57bl/6 마우스(18-21g)를 실험에 이용하였다. 배양한 대장암 세포 MC-38은 5x105 cell을 수확(harvest)한 후, 50㎕의 PBS에서 재부유하여 마우스의 오른쪽 대퇴부 피하에 주입하였다.6-week-old C57bl/6 mice (18-21g) were used in the experiment for the production of the MC-38 cell transplantation animal model. Cultured colon cancer cells MC-38 were harvested (harvest) of 5x10 5 cells, resuspended in 50 μl of PBS, and injected subcutaneously into the right thigh of the mouse.
LLC1 세포 이식 동물모델 제작을 위해서 6주령 C57bl/6 마우스(18-21g)를 실험에 이용하였다. 배양한 대장암 세포 LLC1은 5x105 cell을 수확(harvest)한 후, 50㎕의 PBS에서 재부유하여 마우스의 오른쪽 대퇴부 피하에 주입하였다.6-week-old C57bl/6 mice (18-21g) were used in the experiment for the production of LLC1 cell transplantation animal models. Cultured colorectal cancer cells LLC1 were harvested (harvest) of 5x10 5 cells, resuspended in 50 μl PBS, and injected subcutaneously into the right thigh of the mouse.
B16F10 흑색종 세포 이식 동물모델 제작을 위해서 6주령 C57bl/6 마우스(18-21g)를 실험에 이용하였다. 배양한 대장암 세포 B16F10은 5x105 cell을 수확(harvest)한 후, 50㎕의 PBS에서 재부유하여 마우스의 오른쪽 대퇴부 피하에 주입하였다.6-week-old C57bl/6 mice (18-21g) were used in the experiment for the production of the B16F10 melanoma cell transplantation animal model. The cultured colorectal cancer cells B16F10 were harvested (harvest) of 5x10 5 cells, resuspended in 50 μl of PBS, and injected subcutaneously into the right thigh of the mouse.
4-3. 항종양 효과 분석4-3. Anti-tumor effect analysis
종양이 약 80~100mm3의 부피로 형성된 MC-38 대장암 세포 이식 마우스의 꼬리에 와이셀라 시바리아 LBML2 (Weissella cibaria LBML2) 균주1x109/100 ul를 정맥 내 주사하고, 음성 대조군에는 PBS를 투여하였다. 이후, 시간 경과에 따라 MC-38 대장암 세포 이식 마우스의 대장암 종양 크기의 부피 변화를 측정한 결과를 도 1에 나타내었다.Weissella cibaria LBML2 (Weissella cibaria LBML2) strain 1x10 9 /100 ul was intravenously injected into the tail of MC-38 colorectal cancer cell-transplanted mice in which the tumor was formed in a volume of about 80-100 mm 3 , and PBS was administered to the negative control group. did. Thereafter, the results of measuring the volume change of the colorectal cancer tumor size of the MC-38 colorectal cancer cell transplanted mice over time are shown in FIG. 1 .
도 1에 나타난 바와 같이, 와이셀라 시바리아 LBML2 균주를 투여한 그룹의 경우 12일이 경과한 후, 음성 대조군 대비 대장암 부피 성장이 약 54% 감소하여 현저한 항종양 효과가 나타남을 확인하였다.As shown in FIG. 1 , in the case of the group administered with the Weissella civaria LBML2 strain, after 12 days, it was confirmed that the colorectal cancer volume growth was decreased by about 54% compared to the negative control group, resulting in a significant antitumor effect.
종양이 약 80~100mm3의 부피로 형성된 LLC1 폐암 세포 이식 마우스의 꼬리에 와이셀라 시바리아 LBML2 (Weissella cibaria LBML2) 균주 1x109/100 ul를 정맥 내 주사하고, 음성 대조군에는 PBS를 투여하였다. 이후, 시간 경과에 LLC1 폐암 세포 이식 마우스의 폐암 종양 크기의 부피 변화를 측정한 결과를 도 2에 나타내었다.Weissella cibaria LBML2 (Weissella cibaria LBML2) strain 1x10 9 /100 ul was intravenously injected into the tail of LLC1 lung cancer cell transplantation mice in which the tumor was formed in a volume of about 80-100 mm 3 , and PBS was administered to the negative control group. Thereafter, the results of measuring the volume change of the lung cancer tumor size of LLC1 lung cancer cell transplantation mice over time are shown in FIG. 2 .
도 2에 나타난 바와 같이, 와이셀라 시바리아 LBML2 균주를 투여한 그룹의 경우 6일이 경과한 후, 음성 대조군 대비 폐암 부피 성장이 약 49% 감소하여 현저한 항종양 효과가 나타남을 확인하였다.As shown in Figure 2, in the case of the group administered with the Weissella civaria LBML2 strain, after 6 days, lung cancer volume growth was reduced by about 49% compared to the negative control group, confirming that a significant antitumor effect appeared.
종양이 약 80~100mm3의 부피로 형성된 B16F10 흑색종 세포 이식 마우스의 꼬리에 와이셀라 시바리아 LBML2 (Weissella cibaria LBML2) 균주1x109/100 ul를 정맥 내 주사하고, 음성 대조군에는 PBS를 투여하였다. 이후, 시간 경과에 B16F10 흑색종 세포 이식 마우스의 흑색종 종양 크기의 부피 변화를 측정한 결과를 도 3에 나타내었다.Weissella cibaria LBML2 (Weissella cibaria LBML2) strain 1x10 9 /100 ul was intravenously injected into the tail of B16F10 melanoma cell-transplanted mice in which the tumor was formed in a volume of about 80-100 mm 3 , and PBS was administered to the negative control group. Thereafter, the results of measuring the volume change of the melanoma tumor size of the B16F10 melanoma cell transplanted mice over time are shown in FIG. 3 .
도 3에 나타난 바와 같이, 와이셀라 시바리아 LBML2 균주를 투여한 그룹의 경우 6일이 경과한 후, 음성 대조군 대비 흑색종양 부피 성장이 약 51% 감소하여 현저한 항종양 효과가 나타남을 확인하였다.As shown in FIG. 3 , in the case of the group administered with the Weissella civaria LBML2 strain, it was confirmed that the melanoma volume growth was reduced by about 51% compared to the negative control group after 6 days, showing a significant antitumor effect.
실시예Example 5. 미성숙 수지상 세포의 5. Immature Dendritic Cells 성숙화maturation 유도 및 분화된 수지상 세포의 활성화 확인 Confirmation of activation of induced and differentiated dendritic cells
5-1. 염증성 5-1. inflammatory 사이토Saito 카인 발현 측정 Measuring Cain Expression
마우스의 대퇴골(femur)과 경골(tibia)로의 골수세포(bone marrow cells)로부터 단핵구(monocytes)를 Ficoll gradient 방법을 이용하여 원심분리를 통해 분리하였다. 상기 분리된 단핵구는 2 x 106/well의 농도로 6 well 플레이트에 준비하였으며, 태아 소 혈청(Fetal Bovine Serum, FBS)이 포함된 배양액(RPMI)에 GM-CSF 20 ng/ml, IL-4 10 ng/ml를 함께 처리하였다. 처리 후, 6일 동안 배양하였으며 3일이 경과하였을 때, 신선한 배지와 사이토카인으로 교체해주어 분화된 미성숙 수지상 세포를 수득하였다. 수득한 미성숙 수지상 세포(Immature DC)를 성숙 수지상 세포(mature DC)로의 분화를 위해 2차 자극원인 와이셀라 시바리아 LBML2 균주와 24시간 공동배양하였으며, 수지상 세포의 활성은 realtime PCR을 통해 염증성 사이토카인 TNFa의 발현을 측정하여 확인하였다. 상기 TNFa의 발현 측정 결과를 도 4에 나타내었다.Monocytes were isolated from the bone marrow cells of the mouse femur and tibia by centrifugation using the Ficoll gradient method. The isolated monocytes were prepared in a 6-well plate at a concentration of 2 x 10 6 /well, GM-CSF 20 ng/ml, IL-4 in a culture medium (RPMI) containing Fetal Bovine Serum (FBS) 10 ng/ml were treated together. After treatment, the cells were cultured for 6 days, and when 3 days had elapsed, they were replaced with fresh medium and cytokines to obtain differentiated immature dendritic cells. The obtained immature dendritic cells (Immature DC) were co-cultured with Weissella cibaria LBML2 strain, a secondary stimulator, for 24 hours for differentiation into mature dendritic cells (mature DC). It was confirmed by measuring the expression of TNFa. The results of measuring the expression of TNFa are shown in FIG. 4 .
도 4에 나타난 바와 같이, 와이셀라 시바리아 LBML2 균주와 공동배양한 경우 대조군 또는 종양 항원만 처리했을 경우보다 TNFa의 발현을 크게 증가시킴으로써 수지상 세포의 활성을 현저히 증가시키는 것을 확인할 수 있었다. As shown in FIG. 4 , when co-cultured with the Weissella civaria LBML2 strain, it was confirmed that the activity of dendritic cells was significantly increased by significantly increasing the expression of TNFa than when only the control or tumor antigen was treated.
5-2. In vitro T 세포 활성화 분석(T cell activation assay)5-2. In vitro T cell activation assay
상기 실시예 5-1에서 와이셀라 시바리아 LBML2 균주에 의해 성숙화된 수지상 세포가 T 세포의 분열을 유도할 수 있는지 알아보기 위해 성숙화된 수지상 세포와 T 세포를 3일 동안 공동배양한 후, CCK8 assay를 이용하여 T 세포의 활성 및 증식 정도를 측정한 결과를 도 5에 나타내었다.To determine whether the dendritic cells matured by the Weissella civaria LBML2 strain in Example 5-1 can induce T cell division, the matured dendritic cells and T cells were co-cultured for 3 days, then CCK8 assay 5 shows the results of measuring the activity and proliferation of T cells using
도 5에 나타난 바와 같이, 와이셀라 시바리아 LBML2 균주에 의해 성숙화된 수지상 세포는 T 세포의 분열을 유의미하게 증가시킴으로써, 상기 활성화된 T 세포에 의해 항암 활성이 증가할 것임을 확인할 수 있었다.As shown in FIG. 5 , dendritic cells matured by the Weissella civaria LBML2 strain significantly increased the division of T cells, thereby confirming that the anticancer activity would increase by the activated T cells.
[수탁번호][accession number]
기탁기관명 : 한국미생물보존센터(국외)Name of deposit institution: Korea Microorganism Conservation Center (Overseas)
수탁번호 : KCCM12948PAccession number: KCCM12948P
수탁일자 : 20210203Deposit date: 20210203
Figure PCTKR2022002604-appb-img-000001
Figure PCTKR2022002604-appb-img-000001

Claims (13)

  1. 와이셀라 시바리아 LBML2(Weissella cibaria LBML2, 기탁번호 KCCM12948P) 균주 또는 이의 배양물을 유효성분으로 포함하는 암의 예방 또는 치료용 약학 조성물.Weissella cibaria LBML2 (Weissella cibaria LBML2, accession number KCCM12948P) strain or a pharmaceutical composition for the prevention or treatment of cancer comprising a culture thereof as an active ingredient.
  2. 제1항에 있어서, According to claim 1,
    상기 암은 대장암, 폐암, 흑색종양, 방광암, 유방암, 갑상선암, 부갑상선암, 인후암, 후두암, 식도암, 췌장암, 위암, 설암, 피부암, 뇌종양, 자궁암, 두담낭암, 구강암, 신장암, 간암, 결장암, 직장암, 또는 혈액암인 것을 특징으로 하는 암의 예방 또는 치료용 약학 조성물.The cancers include colorectal cancer, lung cancer, black tumor, bladder cancer, breast cancer, thyroid cancer, parathyroid cancer, throat cancer, laryngeal cancer, esophageal cancer, pancreatic cancer, stomach cancer, tongue cancer, skin cancer, brain tumor, uterine cancer, double gallbladder cancer, oral cancer, kidney cancer, liver cancer, colon cancer, A pharmaceutical composition for preventing or treating cancer, characterized in that it is rectal cancer, or blood cancer.
  3. 제1항에 있어서, According to claim 1,
    상기 조성물은 경구 투여 또는 비경구 투여의 방법으로 투여 가능한 것을 특징으로 하는 암의 예방 또는 치료용 약학 조성물.The composition is a pharmaceutical composition for the prevention or treatment of cancer, characterized in that it can be administered by oral administration or parenteral administration.
  4. 제3항에 있어서,4. The method of claim 3,
    상기 비경구 투여의 방법은 정맥 내 주입, 피하 주입, 근육 주입, 복강 주입, 내피 투여, 국소 투여, 비내 투여, 폐 내 투여 또는 직장 내 투여인 것인, 암의 예방 또는 치료용 약학 조성물.The method of parenteral administration is intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, endothelial administration, topical administration, intranasal administration, intrapulmonary administration or rectal administration, the pharmaceutical composition for the prevention or treatment of cancer.
  5. 제1항에 있어서,The method of claim 1,
    상기 와이셀라 시바리아 LBML2 균주는 생균 또는 사균의 형태인 것을 특징으로 하는 암의 예방 또는 치료용 약학 조성물.The Weissella civaria LBML2 strain is a pharmaceutical composition for preventing or treating cancer, characterized in that it is in the form of live or dead cells.
  6. 제1항에 있어서,The method of claim 1,
    상기 배양물은 생균 배양물 또는 사균 상등액인 것을 특징으로 하는 암의 예방 또는 치료용 약학 조성물.The culture is a pharmaceutical composition for the prevention or treatment of cancer, characterized in that the live cell culture or dead cell supernatant.
  7. 와이셀라 시바리아 LBML2(Weissella cibaria LBML2, 기탁번호 KCCM12948P) 균주 또는 이의 배양물을 유효성분으로 포함하는 암의 예방 또는 개선용 식품 조성물.Weissella cibaria LBML2 (Weissella cibaria LBML2, accession number KCCM12948P) a food composition for preventing or improving cancer comprising a strain or a culture thereof as an active ingredient.
  8. 제7항에 있어서,8. The method of claim 7,
    상기 식품은 건강기능식품인, 암의 예방 또는 개선용 식품 조성물.The food is a health functional food, a food composition for the prevention or improvement of cancer.
  9. 제7항에 있어서,8. The method of claim 7,
    상기 식품은 빵, 떡, 캔디, 초콜릿, 껌, 아이스크림, 우유, 치즈, 식육가공품, 어육가공품, 김치, 간장, 된장, 고추장, 춘장, 청국장, 식초, 케첩, 카레, 드레싱, 음료 및 발효유로 이루어지는 군으로부터 선택되는 어느 하나의 식품인 것을 특징으로 하는 암의 예방 또는 개선용 식품 조성물.The food is bread, rice cake, candy, chocolate, gum, ice cream, milk, cheese, processed meat products, processed fish meat products, kimchi, soy sauce, soybean paste, red pepper paste, chunjang, cheonggukjang, vinegar, ketchup, curry, dressing, beverage and fermented milk consisting of Food composition for the prevention or improvement of cancer, characterized in that any one food selected from the group.
  10. 와이셀라 시바리아 LBML2(Weissella cibaria LBML2, 기탁번호 KCCM12948P) 균주 또는 이의 배양물을 유효성분으로 포함하는 암의 예방 또는 개선용 식품 첨가제 조성물.Weissella cibaria LBML2 (Weissella cibaria LBML2, accession number KCCM12948P) food additive composition for preventing or improving cancer comprising a strain or a culture thereof as an active ingredient.
  11. 와이셀라 시바리아 LBML2(Weissella cibaria LBML2, 기탁번호 KCCM12948P) 균주 또는 이의 배양물을 유효성분으로 포함하는 가축의 암 예방 또는 개선용 사료 조성물.Weissella cibaria LBML2 (Weissella cibaria LBML2, accession number KCCM12948P) a feed composition for preventing or improving livestock cancer comprising a strain or a culture thereof as an active ingredient.
  12. 제11항에 있어서,12. The method of claim 11,
    상기 가축은 돼지, 소, 말, 양, 토끼, 염소, 쥐, 햄스터, 기니피그, 개, 고양이, 닭, 칠면조, 오리, 거위, 꿩, 메추라기, 잉어, 붕어 및 송어로 이루어지는 군으로부터 선택되는 어느 하나인 것인, 가축의 암 예방 또는 개선용 사료 조성물.The livestock is any one selected from the group consisting of pig, cow, horse, sheep, rabbit, goat, rat, hamster, guinea pig, dog, cat, chicken, turkey, duck, goose, pheasant, quail, carp, crucian carp and trout A feed composition for preventing or improving cancer in livestock.
  13. 와이셀라 시바리아 LBML2(Weissella cibaria LBML2, 기탁번호 KCCM12948P) 균주 또는 이의 배양물을 유효성분으로 포함하는 가축의 암 예방 또는 개선용 사료 첨가제 조성물.Weissella cibaria LBML2 (Weissella cibaria LBML2, accession number KCCM12948P) feed additive composition for preventing or improving livestock cancer comprising a strain or a culture thereof as an active ingredient.
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