KR102390755B1 - Composition for prevention or treatment of cancer comprising Weissella paramesenteroides WiKim0137 as active ingredient - Google Patents

Abstract

The present invention relates to a composition for preventing or treating cancer, which comprises a novel Weissella paramesenteroides WiKim0137 strain isolated from kimchi or a culture thereof as an active ingredient. The Weissella paramesenteroides WiKim0137 strain or the culture thereof according to the present invention exhibits excellent anticancer activity and thus can be usefully used as a composition for treating, preventing or alleviating human or animal cancer.

Description

Composition for prevention or treatment of cancer comprising Weissella paramesenteroides WiKim0137 strain as an active ingredient {Composition for prevention or treatment of cancer comprising Weissella paramesenteroides WiKim0137 as active ingredient}

The present invention relates to a novel Weissella paramesenteroides WiKim0137 strain having anticancer activity isolated from kimchi and a composition for preventing or treating cancer comprising the 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. In particular, due to the westernization of dietary habits, high-fat diets are common, and due to the rapid increase in environmental pollutants and the increase in alcohol consumption, colorectal cancer, breast cancer, prostate cancer, etc. are continuously increasing. Lung cancer is on the rise due to the increase in cancer and air pollution. In this situation, there is an urgent need to create anticancer substances that can contribute to the promotion of human health, the improvement of the quality of healthy life, and the promotion of human health by enabling the early prevention and treatment of cancer.

Although the survival rate of cancer patients is increasing due to the development of medical technology, the mortality rate is still high. For example, cancer-related deaths accounted for 27.5% of all deaths in Korea in 2019, still being recorded as the number one cause of death (Statistics Korea). Currently, as a cancer treatment method performed in clinical practice, many methods such as chemotherapy, targeted chemotherapy, and radiation therapy have been tried. However, in cancer treatment, it cannot completely remove all cancer cells and has strong toxicity to normal cells. The reasons include incomplete cancer target, insufficient penetration into cancer tissue, and limited cancer killing effect due to tumor heterogeneity. In fact, due to disordered and incompletely generated blood vessels in cancer tissue, there is an area where therapeutic substances and nutrients are not delivered well, and the cells in the hypoxic/low glucose area generated here stop growing and are resistant to most anticancer drugs or radiotherapy. will have In addition, the rapid growth of cancer cells widens the gap between blood vessels, reduces blood flow, and further decreases drug delivery to cancer tissues far from blood vessels.

Research on cancer treatment using bacterial strains with cancer-targeting ability is increasing worldwide interest, and it has been reported that microorganisms including some lactic acid bacteria grow specifically in a microenvironment created as a tumor. With these characteristics, bacteriotherapy technology, that is, cancer treatment research based on attenuated bacteria is being continuously tried. The use of probiotic bacteria is being considered to solve the toxicity of attenuated bacteria with tumor orientation under existing studies, but in these cases, it was observed that the anticancer effect was low or the tumor selectivity was low. Kimchi lactic acid bacteria-based lactic acid bacteria-related technology with excellent antitumor effects that can overcome the safety and efficiency problems of bacteriotherapy while supplementing the limitations of existing anticancer drugs is rarely reported. Prevention of solid cancer containing lactic acid bacteria with excellent effects And there is a demand for the development of a composition useful for treatment and securing various resources. Lactobacillus is widely distributed in the oral cavity, intestine, vagina, and feces of humans and animals, and is 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.

Currently, Korean Patent Application Laid-Open No. 10-2015-0031922 discloses the immune activity of the Weissella civaria strain, or Korean Patent No. 10-2009742 discloses the anticancer activity of the Weissella civaria strain, but anticancer activity There is no description of the Weissella paramesenteroid strain or a composition comprising the same having a.

Korean Patent Laid-Open No. 10-2015-0031922 Korean Patent Registration No. 10-2009742

It is an object of the present invention to provide a novel Weissella paramesenteroides WiKim0137 strain of accession number KCCM13022P.

In addition, another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer comprising the Weissella paramesenteroides WiKim0137 strain and a culture thereof as an active ingredient.

In addition, another object of the present invention is to provide a food composition or a food additive composition for preventing or improving cancer comprising the Weissella paramesenteroides WiKim0137 strain and a culture thereof as an active ingredient.

In addition, another object of the present invention is to provide a feed composition or feed additive composition for preventing or improving cancer comprising the Weissella paramesenteroides WiKim0137 strain and a culture thereof as an active ingredient.

In addition, another object of the present invention is to provide a lactic acid bacteria starter for fermentation comprising the Weissella paramesenteroides WiKim0137 strain and a culture thereof as an active ingredient.

Accordingly, the present inventors have tried to find a lactic acid bacteria strain having anticancer activity from kimchi, and as a result, a novel Weissella paramesenteroides WiKim0137 strain having anticancer activity in various carcinomas was isolated and identified, and the present invention was completed.

In order to achieve the above object, there is provided a pharmaceutical composition for preventing or treating cancer comprising the Weissella paramesenteroides WiKim0137 strain or a culture thereof as an active ingredient.

In addition, the present invention provides a food composition or food additive composition for preventing or improving cancer comprising Weissella paramesenteroides WiKim0137 strain or a culture thereof as an active ingredient.

In addition, the present invention provides a feed composition or feed additive composition for preventing or improving cancer in livestock comprising Weissella paramesenteroides WiKim0137 strain or a culture thereof as an active ingredient.

In addition, the present invention provides a lactic acid bacteria starter for fermentation comprising the Weissella paramesenteroides WiKim0137 strain or a culture thereof as an active ingredient.

The Weissella paramesenteroides WiKim0137 strain or a culture thereof according to the present invention exhibits excellent anticancer activity against various carcinomas, so it can be usefully used as a composition for use in the treatment, prevention or improvement of human or animal cancer. can

1 is a graph showing the results of measuring the cancer cell killing ability after treatment of the WiKim0137 composition according to the present invention to colorectal cancer CT26 and MC38 cells.
2 is a graph showing the results of measuring the cancer cell killing ability after treatment of the WiKim0137 composition according to the present invention to lung cancer H1650 and A549 cells.
3 is a graph showing the results of measuring the cancer cell killing ability after treating the WiKim0137 composition according to the present invention to pancreatic cancer ASPC1 cells.
4 is a graph showing the results of measuring the cancer cell killing ability after treatment of the WiKim0137 composition according to the present invention to the liver cancer HepG2 cells.
5 is a graph showing the results of measuring the cancer cell killing ability after treating the WiKim0137 composition according to the present invention to normal skin cells CCD986sk.
6 is a graph showing the results of measuring the change in tumor volume over time after administration of the WiKim0137 composition according to the present invention to colorectal cancer CT26 cell transplanted mice.

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. .

As a result of 16S rDNA sequencing analysis for the identification and classification of microorganisms, the strain isolated from kimchi through the examples of the present invention was found to have the nucleic acid sequence of SEQ ID NO: 1 (SEQ ID NO: 1).

Therefore, the microorganism of the present invention having the 16S rDNA base sequence of SEQ ID NO: 1 (SEQ ID NO: 1) was named Weissella paramesenteroides WiKim0137, and was sent to the Korea Microorganism Conservation Center on July 21, 2021. It was deposited on the date (Accession No. KCCM13022P).

The composition comprising the Weissella paramesenteroides WiKim0137 strain of the present invention or a culture thereof as an active ingredient has a preventive or therapeutic effect on cancer and can be used as a pharmaceutical composition.

The Weissella paramesenteroides WiKim0137 strain of the present invention is a lactic acid bacteria strain. The Weissella paramesenteroides WiKim0137 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 Weissella paramesenteroides WiKim0137 have intestinal effect and immune enhancing effect.

The Weissella paramesenteroides WiKim0137 strain of the present invention is a Gram-positive bacterium and facultative anaerobe, and the cells are in the form of cocci. The growth temperature is 20 °C to 40 °C, and the optimum growth temperature is 30 °C. The growth pH is 4.0 to 7.5, and the optimum growth pH is 6.0 to 6.5. In addition, the characteristics confirmed using the API 50 CH kit for identification of lactic acid bacteria were described in the following Examples.

The strain can be used by inoculating 0.1 to 10% in MRS broth and culturing at 25 to 37° C. for 4 to 48 hours.

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.

The cancer is bladder cancer, ovarian cancer, breast cancer, skin cancer, melanoma, thyroid cancer, parathyroid cancer, throat cancer, laryngeal cancer, nasal cancer, salivary gland cancer, esophageal cancer, pancreatic cancer, stomach cancer, tongue cancer, skin cancer, brain tumor, uterine cancer, gallbladder cancer, biliary tract cancer, oral cancer , kidney cancer, pancreatic cancer, colon cancer, rectal cancer, anal cancer, liver cancer, may be any one cancer selected from the group consisting of lung cancer and colon cancer, but is not limited thereto.

The Weissella paramesenteroides WiKim0137 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 Weissella paramesenteroides WiKim0137 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.

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. may be administered, but is not limited thereto.

A suitable dosage of the composition may be prescribed in various ways depending on factors such as formulation method, administration mode, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity of the patient.

When the composition of the present invention is used 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.

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.

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 include sterile and biocompatible, 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.

Furthermore, by using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA by an appropriate method in the field, it can be preferably formulated according to each disease or component.

The composition comprising the Weissella paramesenteroides WiKim0137 strain of the present invention or a culture thereof as an active ingredient may be used as a food composition for preventing or improving cancer or a food additive composition.

The food composition may be in the form of a health functional food.

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 Controlling nutrients for the structure and function of the human body or obtaining useful effects for health purposes such as physiological action (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.

Items listed in the "Food Additives Code", 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, 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 sausages, bacon, etc. Fish and meat products Ramen, dried noodles, raw noodles, fried noodles, luxurious dried noodles, improved soft noodles, frozen noodles, pastas, etc. Fruit drinks, vegetable drinks, carbonated drinks, soy milk, Beverages such as yogurt, lactic acid bacteria, mixed drinks, soy sauce, soybean paste, red pepper paste, chunjang, cheonggukjang, mixed soy sauce, vinegar, sauces, tomato ketchup, curry and seasonings such as dressings, margarine, shortening, and pizza, but limited to this it's not going to be

In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, 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.

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 conventional beverages. 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 (taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. .

In addition, the composition comprising the Weissella paramesenteroides WiKim0137 strain of the present invention or 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.

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 additive is an organic acid such as citric acid, humic acid, adipic acid, lactic acid, malic acid, sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polyphosphate), etc., 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.

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. 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 may 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.

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.

The feed or feed additive of the present invention can be applied to a number of animal diets including mammals, poultry and fish.

Pigs, cows, 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 chickens as the poultry, It can also be used for turkey, duck, geese, pheasant, and quail, and the like, and can be used for carp, crucian carp and trout as the fish, but is not limited thereto.

In addition, the composition comprising the Weissella paramesenteroides WiKim0137 strain of the present invention or a culture thereof as an active ingredient may be used as a starter for fermentation.

Example 1. Isolation and identification of strains

First, after crushing the kakdugi kimchi sample, serial dilution (10 ³ ~ 10 ⁸ ) of the kimchi solution with phosphate buffered saline (PBS) was applied to the primary lactic acid bacteria selection medium, MRS, and then cultured at 30° C. for more than 24 hours. Colonies formed after culture were separated into single colonies and subcultured to select homogeneous colonies. The selected single colonies were collected in a loop, cultured in MRS broth, and then genomic DNA was extracted from the culture medium. In order to confirm the 16S rDNA base sequence, the obtained genomic DNA product was requested for analysis by Genotech. 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). It was named WiKim0137 (Weissella paramesenteroides WiKim0137) strain, and was deposited at the Korea Microorganism Conservation Center on July 21, 2021.

The mycological properties of the Weissella paramesenteroides WiKim0137 strain were confirmed, and the confirmed results are described below.

- Cell type: Bacillus

- Gram staining: positive

- Optimal growth temperature: 30℃

- Growable growth pH 4.0 ~ 7.5

- Optimal pH 6.0 ~ 6.5

- Effect on oxygen: facultative anaerobic

- Catalase: -

- Gas formation: -

- Indole production: -

- Lactic acid production: +

- Hemolysis: -

- Glycolysis pattern using API 50 CH (see Table 1 below)

reaction substrate result reaction substrate result 1 glycerol - 26 you saved - 2 erythritol - 27 Cellobiose - 3D Arabinos - 28 Maltos + 4 L arabinose + 29 Lactose - 5 Ribose D-Ribose + 30 Melibios + 6 D xylose D-xylose + 31 sucrose + 7 L Xylose L-Xylose - 32 Trehalose + 8 Adonitol D-Adonitol - 33 Inulin - 9 methyl-BD-xylofuranocycle - 34 Melesitus - 10 D-galactose + 35 Raffinose - 11 D-glucose + 36 Amidone - 12 D-fructose + 37 glycogen - 13 D-mannose + 38 xylitol - 14 L-Sorbos - 39 Genthiobius - 15 FKASHTM - 40 D Turanos - 16 Dulcitol - 41 D lysos - 17 Inositol - 42 D tagatose - 18 Mannitol - 43 D Fucose - 19 Sorbitol - 44 L fucose - 20 a-methyl-D-mannoside - 45 D Arabitol - 21 a-methyl-D-glucoside + 46 L Arabitol - 22 N-Acetyl-Glucosamine + 47 RMFFNZHSKXP - 23 Amygdalin - 48 2-keto-gluconate - 24 Arbutin - 49 5-keto-gluconate - 25 esculin +

Example 2. Culture of the strain

The isolated Weissella paramesenteroides WiKim0137 strain was inoculated into 25 ml of MRS broth and cultured at 37° C. for 24 hours. After incubation, the culture medium was removed by centrifugation at 6000 rpm for 10 minutes, and the cells were washed three times with a phosphate buffered saline (PBS) solution to remove the remaining medium components.

Example 3. In vitro cancer cells ability to kill check the effect

Cell cytotoxicity was measured by measuring LDH leakage (%) released when cells were killed using an LDH assay kit to observe the anticancer activity of the isolated Weissella paramesenteroides WiKim0137 strain. The cell lines used in the anticancer activity efficacy test were colon cancer CT26 and MC38 cell lines, lung cancer H1650 and A549 cell lines, pancreatic cancer ASPC-1 cell line, liver cancer HepG2 cell line, and normal skin cell line CCD-986sk.

Mouse-derived cancer cells CT26 and MC38 were subcultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin in an incubator at 37° C. and 5% CO ₂ , and cells in the growth phase were used. Cultured CT26 and MC38 cells were treated with 0.25% trypsin-EDTA in an incubator at 37° C. for 3 minutes to detach the cells, and then washed twice with DPBS and prepared at 1 X 10 ⁴ cells/well.

Human-derived cancer cells H1650, A549, ASPC1, and HepG2 were subcultured in RPMI medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin in an incubator at 37° C., 5% CO ₂ and tested using cells in the growth phase. did. The cultured H1650, A549, ASPC1, and HepG2 cells were treated with 0.05% trypsin-EDTA for 3 minutes in an incubator at 37° C. to detach the cells, and then washed twice with DPBS to prepare 1 X 10 ⁴ cells/well.

Human-derived normal skin cells CCD-986sk were tested using growth phase cells while subcultured in RPMI medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37°C and 5% CO2 incubator. The cultured CCD-986sk was treated with 0.05% trypsin-EDTA in an incubator at 37° C. for 3 minutes to detach the cells, washed twice with DPBS, and then prepared at 1 X 10 ⁴ cells/well.

Cancer cell apoptosis (cell viability assay) was treated with MOI3 concentration and then cultured for 72 hours, and absorbance (OD 490 nm) was measured after treatment with LDH assay reagent after incubation. LDH leakage (%) of cells treated with Weissella paramesenteroides WiKim0137 was calculated based on 100% absorbance of cells not treated with Weissella paramesenteroides WiKim0137 The results are shown in FIGS. 1 to 5 .

As shown in Figs. 1 to 5, Weissella paramesenteroides WiKim0137 (Weissella paramesenteroides WiKim0137) was treated based on 100% of each cancer cell not treated with Weissella paramesenteroides WiKim0137. CT26 (mouse colorectal cancer cells) 166.7%, MC38 (mouse colorectal cancer cells) 187.8%, H1650 (human non-small cell lung cancer cells) 201.8%, ASPC1 (human pancreatic cancer cells) 161.5%, HepG2 (human liver cancer cells) was 121.2%, confirming the cancer cell killing ability following the observation of LDH leakage, and the cytotoxicity of the normal cell line, CCD-986-sk (human skin cells) was 84.5, similar to that of the control cell, Weissella paramesenteroy. It was observed that the cells survived about 5.5% better than the cells not treated with Death WiKim0137, so apoptosis of normal skin cells was not observed.

Through the above results, it was confirmed that the composition according to the present invention exhibits an ability to kill cancer cells in colon cancer, lung cancer, pancreatic cancer and liver cancer cells, thereby having an excellent anticancer activity, and has no cytotoxicity in normal cells.

Example 4. Analysis of antitumor effect in colorectal cancer CT26 cell transplantation mouse model

4-1. CT26 cell transplantation cancer animal model production

The experimental animals used in the experiment were supplied with 5-week-old male BALB/c mice (Orient Bio, Korea), and were stabilized for 1 week in an animal breeding room in an SPF environment with room temperature of 20±2℃ and humidity of 55±15%. It was bred for the duration of the experiment. As the feed, a normal pellet feed without antibiotics was supplied, and water was provided at any time.

CT26 colorectal cancer cells were purchased from the American Type Culture Collection ( ATCC ), and 10% FBS (fetal bovine serum) and 1% antibiotics (penicillin/streptomycin) were added to DMEM medium and cultured at 5% CO _{2 and} 37°C conditions. .

Cancer cells cultured for cancer cell transplantation were prepared as 1Х10 ⁵ cells, centrifuged at 1,200 rpm for 5 minutes, washed with phosphate-buffered saline (PBS) to remove medium components, and then resuspended in 50 μl of PBS, subcutaneously in the right subcutaneous tissue of the mouse. was injected into About 10 to 14 days after tumor formation, the tumor volume (mm ³ ) was measured using the formula of 3.14 x (length x height x width)/6.

2. Analysis of antitumor effect

After the tumor volume became about 70-80 mm ³ , Weissella paramesenteroides WiKim0137 (Weissella paramesenteroides WiKim0137) was administered to the tumor-formed mice. Weissella paramesenteroides WiKim0137 (Weissella paramesenteroides WiKim0137) lactic acid bacteria were cultured in MRS medium at 37° C. for 24 hours, then the cells were centrifuged at 8,000 rpm for 10 minutes and washed with PBS to remove the remaining medium components. . 1Х10 ¹⁰ CFU/mL of bacteria was quantified using PBS, and 0.1 ㎖ (1Х10 ⁹ CFU) was administered to the experimental animals by tail intravenous injection once, and PBS was administered to the negative control group. The change in the size of the tumor over time was visually confirmed for 10 to 13 days, and the results of measuring the volume of the tumor tissue in the cancer model mouse are shown in FIG. 6 .

As shown in Figure 6, the present invention by confirming that the volume of the tumor of the CT26 (mouse colorectal cancer) cell transplantation mouse injected with Weissella paramesenteroides WiKim0137 (Weissella paramesenteroides WiKim0137) lactic acid bacteria is reduced by 96.53% compared to the control (PBS). It was verified that the composition according to the method exhibits excellent antitumor effect in an animal model.

Korea Microorganism Conservation Center (Overseas) KCCM13022P 20210721

<110> Korea Food Research Institute <120> Composition for prevention or treatment of cancer comprising Weissella paramesenteroides WikimWiKim0137 as active ingredient <130> KP21-0041 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1302 <212> DNA <213> Weissella paramesenteroides <400> 1 agtggcgaac gggtgagtaa cacgtgggta acctacctct tagcagggga taacatttgg 60 aaacaagtgc taataccgta taataccaac aaccgcatgg ttgttggttg aaagatggtt 120 ctgctatcac taagagatgg acccgcggtg cattagctag ttggtaaggt aacggcttac 180 caaggcaatg atgcatagcc gagttgagag actgatcggc cacaatggga ctgagacacg 240 gcccatactc ctacgggagg cagcagtagg gaatcttcca caatgggcgc aagcctgatg 300 gagcaacgcc gcgtgtgtga tgaagggttt cggctcgtaa aacactgtta taagagaaga 360 acggcactga gagtaactgt tcagtgtgtg acggtatctt accagaaagg aacggctaaa 420 tacgtgccag cagccgcggt aatacgtatg ttccaagcgt tatccggatt tattgggcgt 480 aaagcgagcg cagacggtta tttaagtctg aagtgaaagc cctcagctca actgaggaat 540 ggctttggaa actggatgac ttgagtgcag tagaggaaag tggaactcca tgtgtagcgg 600 tgaaatgcgt agatatatgg aagaacacca gtggcgaagg cggctttctg gactgtaact 660 gacgttgagg ctcgaaagtg tgggtagcaa acaggattag ataccctggt agtccacacc 720 gtaaacgatg agtgctagat gttcgagggt ttccgccctt gagtgtcgca gctaacgcat 780 taagcactcc gcctggggag tacgaccgca aggttgaaac tcaaaggaat tgacggggac 840 ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac gcgaagaacc ttaccaggtc 900 ttgacatccc ttgctaatcc tagaaatagg acgttccctt cggggacaag gtgacaggtg 960 gtgcatggtt gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca 1020 acccttatta ttagttgcca gcatttagtt gggcactcta gtgagactgc cggtgacaaa 1080 ccggaggaag gtggggatga cgtcaaatca tcatgcccct tatgacctgg gctacacacg 1140 tgctacaatg gcatatacaa cgagtcgcca acccgcgagg gtgcgctaat ctcttaaagt 1200 atgtctcagt tcggattgta ggctgcaact cgcctacat aagtcggaat cgctagtaat 1260 cgcggatcag cacgccgcgg tgaatacgtt cccgggtctt gt 1302

Claims (14)

Weissella paramesenteroides WiKim0137 (Weissella paramesenteroides WiKim0137) strain of accession number KCCM13022P. Weissella paramesenteroides WiKim0137 (Weissella paramesenteroides WiKim0137; Accession No. KCCM13022P) A pharmaceutical composition for the prevention or treatment of colon cancer, lung cancer, pancreatic cancer or liver cancer comprising a strain or a culture thereof as an active ingredient. delete 3. The method of claim 2,
The composition is a pharmaceutical composition for preventing or treating colon cancer, lung cancer, pancreatic cancer or liver cancer, characterized in that it can be administered by oral administration or parenteral administration. 5. The method of claim 4,
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, colon cancer, lung cancer, pancreatic cancer or liver cancer prevention or a therapeutic pharmaceutical composition. Weissella paramesenteroides WiKim0137 (Weissella paramesenteroides WiKim0137; Accession No. KCCM13022P) A food composition for preventing or improving colon cancer, lung cancer, pancreatic cancer or liver cancer, comprising a strain or a culture thereof as an active ingredient. delete 7. The method of claim 6,
The food is a health functional food, a food composition for preventing or improving colon cancer, lung cancer, pancreatic or liver cancer. 7. The method of claim 6,
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, gochujang, chunjang, cheonggukjang, vinegar, ketchup, curry, dressing and fermented milk from the group consisting of Food composition for the prevention or improvement of colon cancer, lung cancer, pancreatic cancer or liver cancer, characterized in that any one of the selected foods. Weissella paramesenteroides WiKim0137 (Weissella paramesenteroides WiKim0137; Accession No. KCCM13022P) A food additive composition for preventing or improving colon cancer, lung cancer, pancreatic cancer or liver cancer, comprising a strain or a culture thereof as an active ingredient. Weissella paramesenteroides WiKim0137 (Weissella paramesenteroides WiKim0137; Accession No. KCCM13022P) A feed composition for preventing or improving colon cancer, lung cancer, pancreatic cancer or liver cancer of livestock comprising a strain or a culture thereof as an active ingredient. 12. The method of claim 11,
The livestock is pig, cow, horse, sheep, rabbit, goat, rat, hamster, guinea pig, dog, cat, chicken, turkey, duck, goose, pheasant, quail, carp, any one selected from the group consisting of crucian carp and trout The feed composition for preventing or improving colon cancer, lung cancer, pancreatic cancer or liver cancer in livestock. Weissella paramesenteroides WiKim0137 (Weissella paramesenteroides WiKim0137; Accession No. KCCM13022P) A feed additive composition for preventing or improving colon cancer, lung cancer, pancreatic cancer or liver cancer of livestock comprising a strain or a culture thereof as an active ingredient. Weissella paramesenteroides WiKim0137 (Weissella paramesenteroides WiKim0137; accession number KCCM13022P) lactic acid bacteria starter for fermentation comprising a strain or a culture thereof as an active ingredient.

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