WO2020122476A1 - Composition pharmaceutique comprenant un lactobacillus sakei wikim30 comme principe actif pour la prévention ou le traitement d'un cancer - Google Patents

Composition pharmaceutique comprenant un lactobacillus sakei wikim30 comme principe actif pour la prévention ou le traitement d'un cancer Download PDF

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WO2020122476A1
WO2020122476A1 PCT/KR2019/016627 KR2019016627W WO2020122476A1 WO 2020122476 A1 WO2020122476 A1 WO 2020122476A1 KR 2019016627 W KR2019016627 W KR 2019016627W WO 2020122476 A1 WO2020122476 A1 WO 2020122476A1
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cancer
wikim30
composition
lactobacillus
prevention
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PCT/KR2019/016627
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English (en)
Korean (ko)
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최학종
윤미선
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Korea Food Research Institute KFRI
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Korea Food Research Institute KFRI
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Priority claimed from KR1020190082026A external-priority patent/KR102136772B1/ko
Priority to CN201980081668.4A priority Critical patent/CN113164536A/zh
Priority to CA3122124A priority patent/CA3122124A1/fr
Priority to US17/312,698 priority patent/US20220062355A1/en
Priority to AU2019396986A priority patent/AU2019396986A1/en
Priority to BR112021011293A priority patent/BR112021011293A2/pt
Application filed by Korea Food Research Institute KFRI filed Critical Korea Food Research Institute KFRI
Priority to EP19896691.3A priority patent/EP3895719A4/fr
Priority to SG11202106163RA priority patent/SG11202106163RA/en
Priority to JP2021532434A priority patent/JP2022534150A/ja
Publication of WO2020122476A1 publication Critical patent/WO2020122476A1/fr
Anticipated expiration legal-status Critical
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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
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to a composition for preventing or treating cancer comprising Lactobacillus sakei (Accession No. KCCM11878P) isolated from kimchi as an active ingredient.
  • Cancer has a high mortality rate worldwide and is the most common cause of death after cardiovascular disease in Western societies.
  • colonization, breast cancer, and prostate cancer are continuously increasing due to the rapid increase in environmental pollutants and increased alcohol consumption due to the westernization of eating habits, and the smoking population along with the aging of the population.
  • Lung cancer is increasing due to the increase in and air pollution.
  • lactic acid bacteria are widely distributed in humans and animals such as oral, intestinal, vaginal, fecal, and fermented foods such as kimchi, and are closely related to human and animal health. Lactic acid bacteria show various health promoting effects such as intestinal action, suppression of harmful bacteria, immune regulation, lowering of blood cholesterol, and anti-cancer.
  • Korean Patent Publication No. 10-2015-0068061 discloses anticancer activity of Lactobacillus plantarum PNU (KCCM11352P) or Lactobacillus mesenteroides PNU (KCCM11353P), and registered Korean patent No. 10-1287120 discloses a pharmaceutical composition for treating cancer containing Lactobacillus plantarium DSR CK10 [Accession No.: KFCC-11433P] or Lactobacillus plantarium DSR M2 [Accession No.: KFCC-11432P] as an active ingredient.
  • KFCC-11433P Accesion No.: KFCC-11433P
  • Lactobacillus plantarium DSR M2 Accesion No.: KFCC-11432P
  • An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of cancer comprising the Lactobacillus Sakeai strain as an active ingredient.
  • another object of the present invention is to provide a food composition or a food additive composition for the prevention or improvement of cancer comprising the novel kimchi lactic acid bacteria Lactobacillus Sakeai strain as an active ingredient.
  • Another object of the present invention is to provide a feed composition or feed additive composition for the prevention or improvement of cancer, including the novel kimchi lactic acid bacteria Lactobacillus Sakeai strain as an active ingredient.
  • Another object of the present invention is to provide a method for preventing or treating cancer, wherein the composition is administered to an individual other than a human.
  • the present inventors tried to find a lactic acid bacteria strain that shows the prevention and treatment effect of cancer while showing excellent effects as a probiotic from kimchi, and isolated and identified Lactobacillus sakei, WIKIM30, Lactobacillus sakei, WIKIM30, a lactic acid bacteria strain having anticancer activity.
  • the present invention has been completed.
  • the present invention provides a food composition or food additive composition for preventing or improving cancer comprising Lactobacillus sakeai WIKIM30 (Lactobacillus sakei, WIKIM30, accession number KCCM11878P) or a culture thereof as an active ingredient.
  • the present invention provides a feed composition or feed additive composition for preventing or improving cancer in livestock, comprising Lactobacillus sakei WIKIM30 (Lactobacillus sakei, WIKIM30, accession number KCCM11878P) or a culture thereof as an active ingredient.
  • another object of the present invention provides a method for preventing or treating cancer, wherein the composition is administered to an individual other than a human.
  • Lactobacillus Sakeai WIKIM30 exhibits excellent anticancer activity in animal models and cancer cell lines in which cancer cells are transplanted, it can be usefully used as a composition for the treatment, prevention or improvement of cancer in humans or animals Can be.
  • FIG. 1 is a photograph of a tumor portion of a mouse for observing a change in the size of a tumor cell over time after intravenous injection of a composition according to the present invention to a CT26 cell transplanted mouse once (Single) or three times (Serial) It is a diagram showing.
  • FIG. 2 is a view showing a graph of measuring the change in volume of tumor cells over time after a single (or single) or three (Serial) intravenous injection of CT26 cell-grafted mice according to the present invention.
  • FIG. 3 is a view showing a photograph of a tumor portion of a mouse for observing that the size of tumor cells changes over time after intravenous injection of a composition according to the present invention into a B16F10 skin melanoma mouse.
  • FIG. 4 is a graph showing a change in the volume of tumor cells over time after intravenous injection of a composition according to the present invention into a B16F10 skin melanoma mouse.
  • FIG. 5 is a graph showing the composition of the present invention treated with Lactobacillus Sakeai WIKIM30 in mouse-derived colorectal cancer CT26 cell line, human-derived colorectal cancer SW620 cell line, and HCT116 cell line, and measuring cell viability assay.
  • FIG. 6 is a graph showing the measurement of cell viability assay by treating Lactobacillus Sakeai WIKIM30 in a human-derived non-small lung cancer cell H1650 cell line.
  • FIG. 7 is a graph showing the treatment of Lactobacillus Sakeai WIKIM30 on human-derived pancreatic cancer ASPC1 cell line and PANC1 cell line and measuring cell viability assay.
  • FIG. 8 is a diagram showing a graph of measuring Lactobacillus Sakeai WIKIM30 in a human-derived liver cancer HepG2 cell line and measuring cell viability assay.
  • FIG. 9 is a view showing a graph of measuring Lactobacillus Sakeai WIKIM30 in a human-derived bladder cancer T24 cell line and measuring cell viability assay.
  • FIG. 10 is a diagram showing a graph obtained by treating Lactobacillus Sakeai WIKIM30 on a human-derived normal skin CCD-986-sk cell line and measuring cell viability assay.
  • composition comprising Lactobacillus sakeai WIKIM30 (Lactobacillus sakei, WIKIM30, Accession No. KCCM11878P) or a culture thereof as an active ingredient of the present invention has an effect of preventing or treating cancer and can be used as a pharmaceutical composition.
  • Lactobacillus sakei is a lactic acid bacteria strain derived from kimchi. Although Lactobacillus sakei (WIKIM30) of Lactobacillus sake in the present invention was separated and identified from kimchi, the route for obtaining it is not limited thereto.
  • the microorganism of the present invention having the 16S rRNA nucleotide sequence of SEQ ID NO: 1 was named Lactobacillus sake WIKIM30 (Lactobacillus sakei), and deposited with the Korea Microbiological Conservation Center on September 10, 2015 (Accession No. KCCM11878P).
  • Lactobacillus sakeai WIKIM30 of the present invention is a probiotic, and has a general suit effect and an immune enhancing effect of lactic acid bacteria. It is well known that lactic acid bacteria in the genus Lactobacillus have a formal effect immune enhancing effect.
  • probiotics in the present invention, is understood to mean a living microorganism that has 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 which are living microorganisms with probiotic activity, may have a beneficial effect on the intestinal microflora of a host when fed in the form of dried or fermented products to humans or animals in the form of single or multiple strains.
  • the cancer may include bladder cancer, breast cancer, melanoma, thyroid cancer, parathyroid cancer, rectal cancer, throat cancer, larynx cancer, esophageal cancer, pancreatic cancer, stomach cancer, snow cancer, skin cancer, brain tumor, uterine cancer, bile duct cancer, oral cancer, colon cancer, anal cancer, liver cancer, lung cancer, and It may be any one selected from the group consisting of colorectal cancer, preferably colorectal cancer, skin cancer, lung cancer, pancreatic cancer, liver cancer or bladder cancer, but is not limited thereto.
  • the Lactobacillus Sakeai WIKIM30 included in the composition according to the present invention may exist as a living cell or a living cell, and may also exist in a dried or lyophilized form. Forms and formulation methods of lactic acid bacteria suitable for inclusion in various compositions are well known to those skilled in the art.
  • composition can 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 may be administered. It may be administered, but is not limited thereto.
  • Suitable dosages of the composition can be variously prescribed by factors such as the method of formulation, the mode of administration, the patient's age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion, and response sensitivity.
  • the pharmaceutical composition of the present invention may be prepared using a pharmaceutically acceptable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant includes excipients, boron Release agents, sweeteners, binders, coating agents, expanding agents, lubricants, lubricants or flavoring agents may be used.
  • the pharmaceutical composition may be preferably formulated into a pharmaceutical composition by including one or more pharmaceutically acceptable carriers in addition to the active ingredients described above for administration.
  • the active ingredient can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like.
  • suitable binders, lubricants, disintegrants and colorants can also be included in the mixture.
  • suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, trachocanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzo Eight, sodium acetate, sodium chloride, and the like.
  • Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum, and the like.
  • a composition formulated as a liquid solution sterile and biocompatible, saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol and
  • saline sterile water
  • Ringer's solution buffered saline
  • albumin injection solution dextrose solution
  • maltodextrin solution glycerol
  • ethanol ethanol
  • One or more of these components can be mixed and used, and other conventional additives such as antioxidants, buffers and bacteriostatic agents can be added as necessary.
  • diluents such as aqueous solutions, suspensions, emulsions, pills, capsules, granules or tablets.
  • composition comprising the Lactobacillus sakeai WIKIM30 (Lactobacillus sakei, WIKIM30, Accession No. KCCM11878P) of the present invention or a culture thereof as an active ingredient may be used as a food composition or a food additive composition for preventing or improving cancer.
  • the food composition may be in the form of a health functional food.
  • the “health functional food” refers to food manufactured and processed using ingredients or ingredients with useful functionality in accordance with the Act on Health Functional Food (Article 3, No. 1), and “functional” is It means to control the nutrients for the structure and function of the human body or to obtain useful effects for health purposes such as physiological action (Article 2).
  • the food composition may further include a food additive, and whether or not it is suitable as a "food additive" is the standard for the relevant item and the standard and general test method of the food additives approved by the Korea Food and Drug Administration unless otherwise specified. Judging by the criteria.
  • Items listed in the "Food Additives Fair” include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamon acid, natural additives such as chromosomes, licorice extract, crystalline cellulose, and guar gum, L- And mixed preparations such as sodium glutamate, noodle-added alkalis, preservatives, and tar colorants.
  • chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamon acid
  • natural additives such as chromosomes, licorice extract, crystalline cellulose, and guar gum
  • L- And mixed preparations such as sodium glutamate, noodle-added alkalis, preservatives, and tar colorants.
  • Foods containing the active ingredient of the present invention include breads, rice cakes, dried fruits, candy, chocolates, chewing gum, confectionary ice creams such as confections, ice creams, ice cream products such as ice cream powders, low fat milks, lactose milk, Processed milk, goat milk, fermented milk, butter oil, concentrated milk, milk cream, butter oil, natural cheese, processed cheese, milk powder, dairy products such as whey, meat processed products, egg processed products, meat products such as hamburger fish cake, ham, Fish products such as sausages, bacon, and other processed meat products such as ramen, dried noodles, raw noodles, fried noodles, luxurious dried noodles, improved noodles, frozen noodles, pasta, fruit fruits, vegetable drinks, carbonated drinks, soy milk, Beverages such as lactic acid bacteria drinks such as yogurt, soy sauce, miso, red pepper paste, chunjang, cheonggukjang, mixed vinegar, sauces, tomato ketchup, curry, seasoning foods such as curry, dressing, margarine, shorten
  • 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 adjusting agents, stabilizers, preservatives, glycerin, alcohols, And carbonated agents used in carbonated beverages.
  • composition of the present invention may include flesh for the preparation of natural fruit juice, fruit juice beverages and vegetable beverages. These ingredients can be used independently or in combination.
  • the beverage composition containing the active ingredient of the present invention is not particularly limited to other ingredients, and may contain various flavoring agents or natural carbohydrates, etc. as additional ingredients, such as ordinary drinks.
  • natural carbohydrates described above include monosaccharides (eg, glucose, fructose, etc.); Disaccharides (eg maltose, sucrose, etc.); And polysaccharides, common sugars such as (eg, dextrin, cyclodextrin, etc.), and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • natural flavoring agents such as taumatin and stevia extract (for example, rebaudioside A, glycyrrhizine)
  • synthetic flavoring agents sacharin, aspartame, etc.
  • composition comprising Lactobacillus sakeai WIKIM30 (Lactobacillus sakei, WIKIM30, Accession No. KCCM11878P) or a culture thereof as an active ingredient of the present invention 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, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polymeric phosphate), polyphenols, catechins, alpha-tocopherols, and rosemary Extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, phytinic acid, etc. may further include any one or more of natural antioxidants.
  • the composition When prepared as a feed, the composition may be formulated in a conventional feed form, and may include common feed ingredients.
  • the feed and feed additives include grains, such as crushed or crushed wheat, oats, barley, corn and rice; Vegetable protein feeds, such as feeds based on rape, soybeans, and sunflower; Animal protein feeds, such as blood meal, meat meal, bone meal and fish meal; Sugars and dairy products, for example, may further include dry ingredients made of various milk powders and whey powders, and may further include nutritional supplements, digestion and absorption enhancers, growth promoters, and the like.
  • grains such as crushed or crushed wheat, oats, barley, corn and rice
  • Vegetable protein feeds such as feeds based on rape, soybeans, and sunflower
  • Animal protein feeds such as blood meal, meat meal, bone meal and fish meal
  • Sugars and dairy products for example, may further include dry ingredients made of various milk powders and whey powders, and may further include nutritional supplements, digestion and absorption enhancers, growth promoters, and the like.
  • the feed additive may be administered to animals alone or in combination with other feed additives in an edible carrier.
  • the feed additives can be easily administered to animals as topdressing or by mixing them directly into animal feed or in a separate oral formulation from feed.
  • a pharmaceutically acceptable edible carrier can be solid or liquid, for example corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol.
  • the feed additive can be a tablet, capsule, powder, troche or sugar-coated tablet or a top dressing in undispersed form.
  • the feed additive may be a gelatin soft capsule, or a syrup or suspension, emulsion, or solution formulation.
  • the feed and feed additives may contain adjuvants, such as preservatives, stabilizers, wetting or emulsifying agents, solution accelerators, and the like.
  • the feed additive may be used by adding to animal feed by acupuncture, spraying, or mixing.
  • the feed or feed additive of the present invention is applicable to a number of animal diets including mammals, poultry and fish.
  • the mammal As the mammal, it can be used for pigs, cows, horses, sheep, rabbits, goats, rodents and experimental animals such as rats, hamsters, guinea pigs, pets (eg dogs, cats), and chickens as the poultry. It can also be used for turkey, duck, goose, pheasant, quail, and the like, and may be used as carp, crucian carp and trout, but is not limited thereto.
  • composition comprising Lactobacillus sakeai WIKIM30 (Lactobacillus sakei, WIKIM30, Accession No. KCCM11878P) or a culture thereof as an active ingredient of the present invention may provide a method for preventing or treating cancer administered to an individual except humans. .
  • Lactobacillus Sakeai WIKIM30 (Accession No. KCCM11878P) was pre-sale from the World Kimchi Research Institute Microbial Gene Bank.
  • a single colony of pre-sold Lactobacillus Sakeai WIKIM30 was passaged in MRS agar medium for 48 hours at 30° C., and the cultured single colony was inoculated in 10 ml of MRS liquid medium and shake cultured at 200 rpm for 24 hours at 37° C. . After incubation, the cells were centrifuged at 8,000 rpm for 5 minutes to remove the culture medium, and washed with PBS (Phosphate Buffered Saline) solution three times to remove the remaining medium components.
  • PBS Phosphate Buffered Saline
  • the experimental animals used in the experiment were supplied with a male 5 week old BALB/c mouse and a C57BL/6 mouse (Orient Bio, Korea), and an animal breeding room in an SPF environment where the room temperature was maintained at 20 ⁇ 2°C and humidity 55 ⁇ 15%. After a 1-week stabilization period, they were bred during the experimental period. The feed was fed with a general pellet feed without antibiotics, and water was allowed to be consumed from time to time. All animal breeding, experiments and euthanasia were conducted according to the protocol approved by the Animal Experimental Ethics Committee of the World Kimchi Research Institute. The observation of tumor size change was performed by measuring the volume of the tumor (mm 3 ) using the formula 3.14 x (length x height x width)/6.
  • CT26 mouse colon cancer cells (Korea Cell Line Bank, Korea) were purchased and used. CT26 mouse colon cancer cells were cultured in 5% CO 2 and 37° C. conditions using DMEM medium (Hyclone, USA) containing 10% fetal calf serum and 1% penicillin-streptomycin.
  • CT26 cell transplant animal model 6-week-old BALB/c mice (18-21 g) were used in the experiment.
  • Cultured mouse colon cancer cells CT26 were harvested 1x10 5 cells, resuspended in 50 ⁇ l PBS, and injected under the right thigh of the mouse.
  • the tail of a CT26 cell transplanted mouse with tumors formed in a volume of about 80-100 mm 3 was intravenously injected with Lactobacillus Sakeai WIKIM30.
  • a composition containing Lactobacillus Sakeai WIKIM30 in the CT26 cell transplanted mouse was prepared using PBS to prepare 1 ⁇ 10 9 CFU/0.1ml and 5 ⁇ 10 8 CFU/0.1ml.
  • the tumor-implanted mice were injected intravenously once or three times, and PBS was administered to the negative control group.
  • the results of visually confirming the change in tumor size of colorectal cancer in CT26 cell transplanted mice over time are shown in FIG. 1.
  • the results of measuring the volume change of tumor size in CT26 cell transplanted mice over time are shown in FIG. 2.
  • Example 5 Tumor size change of mouse skin melanoma B16F10 cells
  • B16F10 mouse melanoma cells (Korea Cell Line Bank, Korea) were purchased and used. B16F10 mouse melanoma cells were cultured at 5% CO 2 and 37° C. using DMEM medium (Hyclone, USA) containing 10% fetal calf serum and 1% penicillin-streptomycin.
  • mice For the production of the B16F10 cell transplant animal model, 6-week-old C57BL/6 mice (18-21 g) were used in the experiment.
  • the cultured mouse melanoma cells B16F10 were harvested from each 1x10 5 cell, resuspended in 50 ⁇ l PBS, and injected under the right thigh of the mouse.
  • the tail of a mouse transplanted with B16F10 cells formed with a tumor volume of about 80-100 mm 3 was intravenously injected with Lactobacillus Sakeai WIKIM30.
  • Lactobacillus Sakeai WIKIM30 was prepared by quantifying 1 ⁇ 10 10 CFU/mL bacteria using PBS, 0.1 mL (1 ⁇ 10 9 CFU) was injected intravenously into the experimental animals, and PBS was administered to the negative control group.
  • Fig. 3 shows the results of visually confirming the change in colorectal cancer tumor size of B16F10 cell transplanted mice over time.
  • Cell viability assay was measured using Cell counting Kit-8.
  • Cells used for the anticancer activity efficacy test include mouse-derived colon cancer CT26 cell line, human-derived colon cancer SW620 and HCT116 cell lines, human-derived non-small cell lung cancer H1650 cell line, human-derived pancreatic cancer ASPC1 and PANC1 cell lines, human-derived liver cancer HepG2 cell line, and human-derived bladder cancer The T24 cell line and the normal skin cell line CCD-986-sk cell line were used.
  • the mouse-derived colorectal cancer CT26 cell line was passaged at 5% CO 2 and 37° C. using DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. It was tested using.
  • the cultured B16F10 cells were treated with 0.25% trypsin-EDTA in the incubator at 37°C for 3 minutes to detach the cells, washed twice with DPBS, and prepared at 1 X 10 4 cells/well.
  • SW620, HCT116, H1650, ASPC1, PANC1, HepG2 and T24 cell lines are 5% CO 2 using RPMI medium containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. , it was tested using the growth phase cells subcultured at 37 °C condition. Cultured SW620, HCT116, H1650, ASPC1, PANC1, HepG2 and T24 cells were treated with 0.25% trypsin-EDTA for 3 minutes at 37°C in the incubator, detached and washed twice with DPBS, then 1 X 10 4 cells/well Prepared.
  • FBS fetal bovine serum
  • Human-derived normal skin cell CCD-986-sk cell line was passaged at 5% CO 2 at 37°C using RPMI medium containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. Cultured and tested using cells in the growth phase. The cultured CCD-986-na cells were treated with 0.25% trypsin-EDTA in the incubator at 37°C for 3 minutes to detach the cells, washed twice with DPBS, and prepared at 1 X 10 4 cells/well.
  • FBS fetal bovine serum
  • the cell growth rate (cell viability assay) was treated with MOI1 concentration, and then cultured for 72 hours. After incubation, the cell counting Kit-8 reagent was treated and absorbance (OD 450 nm) was measured. Based on the absorbance of cells not treated with Lactobacillus Sakeai WIKIM30 (comtrol), the growth rate of cells treated with Lactobacillus Sakeai WIKIM30 was calculated, and the experimental results are described below.
  • a cell growth rate of 59.1% was measured in a human-derived non-small cell lung cancer H1650 cell line treated with Lactobacillus Sakeai WIKIM30. From the above results, it can be confirmed that in the case of the lung cancer cell line treated with Lactobacillus Sakeai WIKIM30, since the cell growth rate is significantly inhibited, it has excellent anticancer activity efficacy against lung cancer.
  • a cell growth rate of 18.8% was measured in a human-derived liver cancer HepG2 cell line treated with Lactobacillus Sakeai WIKIM30. From the above results, it can be confirmed that in the case of a liver cancer cell line treated with Lactobacillus Sakeai WIKIM30, cell growth rate is remarkably inhibited and thus has excellent anticancer activity efficacy against liver cancer.
  • a cell growth rate of 53.9% was measured in a bladder cancer T24 cell line derived from humans treated with Lactobacillus Sakeai WIKIM30. From the above results, it can be seen that in the case of the bladder cancer cell line treated with Lactobacillus Sakeai WIKIM30, the cell growth rate is significantly inhibited, and thus it has excellent anticancer activity efficacy against bladder cancer.
  • the two-way t-test determined statistically significant differences in tumor growth between the negative control group and the Lactobacillus Sakeai WIKIM30 treatment group. P ⁇ 0.05 was considered significantly for all tumor growth graph analysis.

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Abstract

La présente invention concerne un nouveau Lactobacillus sakei WIKIM30 (numéro d'accès KCCM11878P) isolé à partir de Kimchi, ainsi qu'une composition le comprenant comme principe actif. Grâce à une excellente activité anticancéreuse dans des modèles animaux dans lesquels des cellules cancéreuses ont été transplantées et des lignées cellulaires cancéreuses, le Lactobacillus sakei WIKIM30 selon la présente invention peut être utilisé avantageusement comme composition pour le traitement, la prévention ou l'atténuation de symptômes d'un cancer chez l'homme ou l'animal.
PCT/KR2019/016627 2018-12-10 2019-11-28 Composition pharmaceutique comprenant un lactobacillus sakei wikim30 comme principe actif pour la prévention ou le traitement d'un cancer Ceased WO2020122476A1 (fr)

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JP2021532434A JP2022534150A (ja) 2018-12-10 2019-11-28 ラクトバチルス・サケイｗｉｋｉｍ３０を有効成分として含む癌の予防又は治療用薬学組成物
CA3122124A CA3122124A1 (fr) 2018-12-10 2019-11-28 Composition pharmaceutique comprenant un lactobacillus sakei wikim30 comme principe actif pour la prevention ou le traitement d'un cancer
US17/312,698 US20220062355A1 (en) 2018-12-10 2019-11-28 Pharmaceutical composition comprising lactobacillus sakei wikim30 as active ingredient for prevention or treatment of cancer
AU2019396986A AU2019396986A1 (en) 2018-12-10 2019-11-28 Pharmaceutical composition comprising Lactobacillus sakei WIKIM30 as active ingredient for prevention or treatment of cancer
BR112021011293A BR112021011293A2 (pt) 2018-12-10 2019-11-28 Composição farmacêutica para prevenção ou tratamento de câncer compreendendo lactobacillus sakei wikim30 como ingrediente ativo
CN201980081668.4A CN113164536A (zh) 2018-12-10 2019-11-28 包含沙克乳酸杆菌wikim30作为有效成分的用于预防或治疗癌症的药学组合物
EP19896691.3A EP3895719A4 (fr) 2018-12-10 2019-11-28 Composition pharmaceutique comprenant un lactobacillus sakei wikim30 comme principe actif pour la prévention ou le traitement d'un cancer
SG11202106163RA SG11202106163RA (en) 2018-12-10 2019-11-28 Pharmaceutical composition for prevention or treatment of cancer comprising lactobacillus sakei wikim30 as active ingredient

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KR1020190082026A KR102136772B1 (ko) 2018-12-10 2019-07-08 락토바실러스 사케아이 wikim30을 유효성분으로 포함하는 암의 예방 또는 치료용 약학 조성물
KR10-2019-0082026 2019-07-08

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