WO2012021016A9 - Souche de lactobacillus ayant une activité anticancéreuse et une composition d'activité anticancéreuse comprenant celle-ci en tant que substance active - Google Patents

Souche de lactobacillus ayant une activité anticancéreuse et une composition d'activité anticancéreuse comprenant celle-ci en tant que substance active Download PDF

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Publication number
WO2012021016A9
WO2012021016A9 PCT/KR2011/005916 KR2011005916W WO2012021016A9 WO 2012021016 A9 WO2012021016 A9 WO 2012021016A9 KR 2011005916 W KR2011005916 W KR 2011005916W WO 2012021016 A9 WO2012021016 A9 WO 2012021016A9
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WIPO (PCT)
Prior art keywords
cancer activity
cells
anticancer
active ingredient
lactobacillus
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Ceased
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PCT/KR2011/005916
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English (en)
Korean (ko)
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WO2012021016A2 (fr
WO2012021016A3 (fr
Inventor
박진형
박기범
김희정
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Publication of WO2012021016A3 publication Critical patent/WO2012021016A3/fr
Publication of WO2012021016A9 publication Critical patent/WO2012021016A9/fr
Anticipated expiration legal-status Critical
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    • 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
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • 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
    • A61K2035/11—Medicinal preparations comprising living procariotic cells
    • A61K2035/115—Probiotics
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus
    • C12R2001/245—Lactobacillus casei

Definitions

  • the present invention was found that Lactobacillus bacteria among the various bacteria isolated during the identification of food-derived fermentation strain having an apple lactic acid bacteria fermentation ability has anti-cancer activity, which leads to the present invention, Lactobacillus strains having anticancer activity and the active ingredient It relates to an anticancer agent comprising.
  • the human body forms the intestinal microflora by various microorganisms.
  • these microorganisms are not only beneficial microorganisms known to be useful for host animals such as lactic acid bacteria, but also microorganisms such as Escherichia coli, Salmonella, Staphylococcus aureus, which show direct or potential hazards to the host.
  • Probiotics are isolated and formulated beneficial microorganisms that live in the intestine of humans or other animals. Aerobic bacteria, anaerobic bacteria, lactic acid bacteria and yeasts are used, but mainly lactic acid bacteria are used.
  • lactic acid bacteria probiotics can maintain the stable intestinal flora by inhibiting abnormal fermentation of harmful microorganisms in the intestine without causing side effects that can occur due to antibiotic misuse, and can prevent diseases caused by infection of harmful microorganisms in advance. It has also been shown to be effective in preventing cancer.
  • the lactic acid bacteria preparation health supplements of today require functions such as anticancer effect, excellent intestinal fixation ability, blood cholesterol reduction action, and immune activity enhancement action.
  • the anticancer effects of lactic acid bacteria include inhibition of cancer cells that induce the production of carcinogens, inhibition of carcinogens and chemicals that become carcinogens.
  • the problem to be solved by the present invention is to establish a strain of lactic acid bacteria excellent in anticancer activity and to provide a drug or a dietary supplement with anticancer activity using the same.
  • the present invention provides an accession number KCCM11072P Lactobacillus casei ( Latobacillus casei ) having anticancer activity.
  • the present invention provides an anticancer active composition comprising Accession No. KCCM11072P Lactobacillus casei as an active ingredient having anticancer activity.
  • Accession No. KCCM11072P Lactobacillus casei having the anticancer activity may be included in the form of live or dead bacteria, and the anticancer active composition may be manufactured and consumed for distribution in pharmaceuticals or dietary supplements. Can be.
  • accession number KCCM11072P Lactobacillus casei has an excellent anticancer effect on gastric cancer and colorectal cancer, and it can be seen that it shows an excellent effect on other solid cancers. Therefore, by using this as an active ingredient can be made without drugs of side effects without having a good anti-cancer activity.
  • Figure 1 shows the results of the MTT assay for gastric cancer cells (KATO-3) of the sample containing Accession No. KCCM11072P Latobacillus casei according to the present invention.
  • Figure 2 shows the immunofluorescence results for KATO-3 cells gastric cancer cell death of the sample containing Accession No. KCCM11072P Latobacillus casei according to the present invention.
  • Figure 3 shows the luciferase assay results for KATO-3 gastric cancer cells of the sample containing Accession No. KCCM11072P Latobacillus casei according to the present invention
  • 3x-kb is the expression of NF-kB
  • PG-13 shows expression of p53.
  • Figure 4 is a view showing the inhibitory effect on gastric cancer transplanted to nude mice of the sample containing the accession number KCCM11072P Latobacillus casei according to the present invention.
  • Figure 5 illustrates the results of the MTT assay for human colon cancer cells (HT-29) of the sample containing the accession number KCCM11072P Latobacillus casei according to the present invention.
  • Lactobacillus bacteria among the various isolates have anticancer activity and established as this strain and deposited with the accession number KCCM11072P, and the deposited strain Samples were tested for anticancer activity by culturing and tested for gastric and colon cancers.
  • the anticancer activity test is described below regarding the preparation of samples and tests for gastric and colorectal cancer.
  • the precipitate corresponding to the cells is washed with PBS solution having a volume of 50 times the weight of the cells, and then centrifuged and washed three times with a culture or metabolite. Cell samples with minimal impact were lyophilized and used for testing.
  • MRS broth medium was used as the culture medium, but Tryptic soy broth medium may be used.
  • KATO-3 stomach cancer cell line
  • 5 X 10 3 cells were incubated for 24 hours.
  • the sample was dissolved in distilled water, and the concentration was treated at 1mg / ml, 0.5mg / ml, 0.25mg / ml, 0.125mg / ml, 0.0625mg / ml, and 24 hours and 48 hours.
  • control group no treat is the control group
  • treatment for each concentration three times was repeated in three groups.
  • Each plate was reacted for 1 hour by adding MTT reagent so that the ratio of the culture medium and the MTT reagent was about 10: 1 after 24 hours and 48 hours. Absorbance was measured at 450 nm with a spectrophotometer, and the average absorbance of 9 wells was used as a result.
  • Reagents used in the MTT assay were DOJINDO's CCK-8 (Cell counting kit-8).
  • Hoechst 33258 (sigma), a fluorescent dye, observes the degree of cell damage and can distinguish apoptotic and non-apoptotic cells by nuclear staining in living cells.
  • KATO-3 cells were treated with samples, washed with PBS, and placed in 3.7% formaldehyde (formaldehyde) for 5 minutes at room temperature to fix the cells.
  • the nucleus and morphology of the cells were observed by confocal observation using a UV filter.
  • Cells were transfected with luciferase-labeled 3x-kB and PG-13 and then treated with samples. This experiment was performed using promega's dual luciferase assay kit.
  • Luminometer was used to measure the degree of luminescence of luciferase and renila luciferase (control), and luciferase / renila luciferase was used as a result.
  • KATO3 cells were injected subcutaneously in the rear of the leg of nude mice. After 19-21 days the mice were sacrificed and tumor tissue was extracted. Experimental treatment started when the tumor grew to an average of 60 mm 3 . Tumors were fixed in PBS containing 4% paraformaldehyde and stained with hematoxylin and eosin. Animal testing was conducted in accordance with the Laboratory Animal Care Regulations of Korea Basic Science Institute.
  • the cell death inducing ability of the sample was measured by analyzing the effect on the cell type and MTT assay.
  • the gastric cancer cells (KATO-3) were observed 24 hours and 48 hours after the extract was treated under a phase contrast microscope, and the results are shown in FIG. 1.
  • the concentration of 500 ug / ml of the sample is IC 50 for gastric cancer cells (KATO-3).
  • the percentage of dead cells of KATO-3 and the morphology of cell death were measured by Annexin V staining, followed by the number of cells.
  • Annexin V staining was used to determine the percentage of cells killed in medium exposed to 500 ug / ml sample.
  • NF-kB is a key transcription factor that is thought to play an important role in carcinogenesis. It regulates several important signaling pathways that promote tumor formation, and its activity is associated with an anti-apoptotic effect. It is the mediator of cellular responses to damage and other stimuli
  • mice were randomly divided into two groups, one group treated with samples and the other group treated with saline.
  • results are shown in FIG. 4, and the results of FIG. 4 show that tumor growth was small in tissues injected at a concentration of 500 ug / ml but normal in tumor-injected tissues. This result further supports that the sample inhibits tumor formation.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Epidemiology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Nutrition Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Biomedical Technology (AREA)
  • Polymers & Plastics (AREA)
  • Food Science & Technology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

La présente invention est basée sur la découverte d'une activité anticancéreuse dans Lactobacillus de nombreux types de bactéries isolées en observant des souches de fermentation d'origine alimentaire ayant la capacité à fermenter Lactobacillus dans des pommes, et concerne Lactobacillus casei (numéro d'accession KCCM11072) ayant une activité anticancéreuse et la composition d'activité anticancéreuse comprenant celle-ci en tant que substance active, qui peut être utilisée en tant que fourniture médicale ou supplément alimentaire de santé.
PCT/KR2011/005916 2010-08-12 2011-08-11 Souche de lactobacillus ayant une activité anticancéreuse et une composition d'activité anticancéreuse comprenant celle-ci en tant que substance active Ceased WO2012021016A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020100077908A KR20120015644A (ko) 2010-08-12 2010-08-12 항암 활성을 갖는 락토바실러스 균주 및 이를 활성성분으로 포함하는 항암 활성 조성물
KR10-2010-0077908 2010-08-12

Publications (3)

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WO2012021016A2 WO2012021016A2 (fr) 2012-02-16
WO2012021016A3 WO2012021016A3 (fr) 2012-05-10
WO2012021016A9 true WO2012021016A9 (fr) 2012-06-28

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WO (1) WO2012021016A2 (fr)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101910808B1 (ko) * 2016-11-28 2018-10-24 주식회사 쎌바이오텍 유산균 유래 p8 단백질 및 이의 항암 용도
KR102680587B1 (ko) * 2018-05-28 2024-07-03 주식회사 쎌바이오텍 유산균 유래 단백질의 활성 단편 펩타이드 및 이의 용도
KR102457422B1 (ko) * 2022-05-04 2022-10-21 주식회사 메디오젠 프로바이오틱스 혼합균주를 유효성분으로 포함하는 대장암의 예방 또는 치료용 조성물
CN116948916B (zh) * 2023-08-11 2024-06-11 四川大学 一种干酪乳杆菌及其应用

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* Cited by examiner, † Cited by third party
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KR100277348B1 (ko) * 1998-12-03 2001-01-15 이은선 겨우살이를 발효시키는 신규 유산균인 락토바실러스 카제이 케이 와이 22713
KR100411198B1 (ko) * 2001-03-13 2003-12-18 매일유업주식회사 암 세포주 증식 억제제
CA2364249A1 (fr) * 2001-11-27 2003-05-27 Unknown-Inconnu Effet de bacteries lactiques sur l'apoptose cellulaire de tumeur
KR20100008831A (ko) * 2008-07-17 2010-01-27 주식회사 쎌바이오텍 락토바실러스 카제이 배양물을 유효성분으로 함유하는항암용 조성물

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WO2012021016A2 (fr) 2012-02-16
KR20120015644A (ko) 2012-02-22
WO2012021016A3 (fr) 2012-05-10

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