[go: up one dir, main page]

WO2020180037A1 - Composition pour la culture d'un mimétique du microbiome intestinal humain, procédé de culture d'un mimétique du microbiome intestinal humain à l'aide d'un système de culture anaérobie, agent microbien préparé à l'aide de ce dernier, et son utilisation - Google Patents

Composition pour la culture d'un mimétique du microbiome intestinal humain, procédé de culture d'un mimétique du microbiome intestinal humain à l'aide d'un système de culture anaérobie, agent microbien préparé à l'aide de ce dernier, et son utilisation Download PDF

Info

Publication number
WO2020180037A1
WO2020180037A1 PCT/KR2020/002710 KR2020002710W WO2020180037A1 WO 2020180037 A1 WO2020180037 A1 WO 2020180037A1 KR 2020002710 W KR2020002710 W KR 2020002710W WO 2020180037 A1 WO2020180037 A1 WO 2020180037A1
Authority
WO
WIPO (PCT)
Prior art keywords
composition
bacteria
microbial community
culture
firmicutes
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/KR2020/002710
Other languages
English (en)
Korean (ko)
Inventor
신재호
김은수
최승대
이유정
박창언
손현우
조영재
조형우
김민철
정연균
박민규
박영준
강기웅
김민지
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Industry Academic Cooperation Foundation of KNU
Original Assignee
Industry Academic Cooperation Foundation of KNU
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from KR1020190024957A external-priority patent/KR20200106419A/ko
Application filed by Industry Academic Cooperation Foundation of KNU filed Critical Industry Academic Cooperation Foundation of KNU
Publication of WO2020180037A1 publication Critical patent/WO2020180037A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • 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, e.g. protozoa; 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

Definitions

  • the present inventors were searching for methods having the same microbial composition as the original feces by simulating and culturing the intestinal microbial community in vitro, while intestinal microbial communities including bacterioidetes, permicutes, proteobacteria, fusobacteria and actinobacteria, etc.
  • the present invention was completed based on this, because it was found a method of culturing similarly to that of Wondyeon, and it was confirmed that the intestinal microbial community obtained therefrom has a therapeutic effect on intestinal diseases.
  • Another object of the present invention is to prepare a microbial community suspension from the feces of a subject; Inoculating the suspension into the composition; And it is to provide a human intestinal microbial community simulation culture method comprising the step of culturing the composition.
  • Another aspect of the present invention provides a microbial preparation comprising as an active ingredient at least one selected from the group consisting of a culture of a microbial community simulated by the above method, a concentrate of the culture, and a dried product of the culture.
  • FIG. 16 shows (a) Bifidobacteriaceae and (b) Akermensi Asia (Bifidobacteriaceae) found in the intestine of mice after administration of a population of intestinal microorganisms simulated by a method according to an embodiment of the present invention.
  • Akkermensiaceae) is a graph showing the family (family) microorganisms.
  • the composition may further include one or more components selected from the group consisting of water-soluble starch, sodium pyruvate, and L-arginine.
  • the composition may further include one or more components selected from the group consisting of Pancreatic digest of casein and Papain digest of soybean.
  • the medium of the present invention is a fastidious anaerobe comprising a brain-heart infusion media (BHI) containing a bovine brain extract, a bovine heart extract and proteose peptone as an active ingredient, starch, sodium pyruvate and L-arginine as an active ingredient.
  • BHI brain-heart infusion media
  • FAB tryptic soy broth
  • pancreatic digests of casein and papain digests of soybeans as active ingredients is more useful for simulated culture in which the ratio of human intestinal microbial communities is maintained.
  • the first step of the present invention is to prepare a microbial community suspension from feces.
  • the step of culturing may be performed for 24 to 120 hours.
  • the microbial community of the present invention may be, for example, a human-derived intestinal microbial community that is normally selected by verification of alpha diversity, beta diversity and/or relative abundance based on criteria commonly applied in the art.
  • At least 120 ⁇ 27g or more of stool samples were obtained per sample from the donor selected in Example 1-1, and some of the obtained stool samples were stored frozen at -80°C, and the extra sample contained 20% glycerin. It was suspended 10 times by weight in 0.85% physiological saline. As a result of measuring the number of viable cells in each stool sample using a hemocytometer, it was confirmed that each stool sample contained about 10 9 to 10 11 intestinal microorganisms per gram. Fecal samples suspended in a buffer solution were stored frozen at -80°C until use in the experiment. The fecal sample pretreatment process was performed in the anaerobic chamber under conditions of 0 ppm to 40 ppm of oxygen and 10000 ppm to 50000 ppm of hydrogen.
  • Example 3 Simulated culture and formulation of fecal samples
  • At least one buffer solution was selected from a buffer group consisting of 0.85% physiological saline and 50% glycerin containing, and mixed 1 to 1 with the BHI medium culture of the simulated microbial community, and formulated in a liquid form (Fig. 7). .
  • DNA was extracted using QIAmp Power Fecal DNA Pro Kit (QIAGEN, Germany) according to a conventional method in the art.
  • the original stool sample suspension and the simulated cultured human intestinal microbial community culture solution of Example 1 were thawed and then 1.8 ml was taken, which was concentrated 6 times by centrifugation, and then extracted according to the manufacturer's instructions.
  • the V4-V5 region of the 16S rRNA gene was specifically amplified by the polymerase chain reaction method using the primers in Table 7.
  • Example 7 Confirmation of the therapeutic effect of inflammatory bowel disease of the simulated cultured intestinal microbial community culture formulation

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

La présente invention concerne une composition pour la culture d'un mimétique du microbiome intestinal humain, un procédé de culture d'un mimétique du microbiome intestinal humain à l'aide d'un système de culture anaérobie, un agent microbien préparé à l'aide de ce dernier, et une utilisation correspondante. Un microbiome obtenu selon le procédé de la présente invention présente une efficacité similaire à celle des micro-organismes intestinaux contenus dans les selles de populations normales, ce qui le rend efficace pour être utilisé dans divers domaines tels que l'amélioration des micro-organismes intestinaux et la préparation d'aliments fonctionnels et cosmétiques.
PCT/KR2020/002710 2019-03-04 2020-02-25 Composition pour la culture d'un mimétique du microbiome intestinal humain, procédé de culture d'un mimétique du microbiome intestinal humain à l'aide d'un système de culture anaérobie, agent microbien préparé à l'aide de ce dernier, et son utilisation Ceased WO2020180037A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2019-0024957 2019-03-04
KR10-2019-0024956 2019-03-04
KR1020190024957A KR20200106419A (ko) 2019-03-04 2019-03-04 동물 장내 미생물 군집의 모사 배양 방법
KR20190024956 2019-03-04

Publications (1)

Publication Number Publication Date
WO2020180037A1 true WO2020180037A1 (fr) 2020-09-10

Family

ID=72337299

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2020/002710 Ceased WO2020180037A1 (fr) 2019-03-04 2020-02-25 Composition pour la culture d'un mimétique du microbiome intestinal humain, procédé de culture d'un mimétique du microbiome intestinal humain à l'aide d'un système de culture anaérobie, agent microbien préparé à l'aide de ce dernier, et son utilisation

Country Status (1)

Country Link
WO (1) WO2020180037A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11679136B2 (en) 2016-05-25 2023-06-20 Mybiotics Pharma Ltd. Composition and methods for microbiota therapy
US11680257B2 (en) 2015-05-11 2023-06-20 Mybiotics Pharma Ltd. Systems and methods for growing a biofilm of probiotic bacteria on solid particles for colonization of bacteria in the gut

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR19990075377A (ko) * 1998-03-20 1999-10-15 유주현 신규한 호염성 단백질 분해효소 및 그를 생산하는 미생물
KR20120121100A (ko) * 2011-04-26 2012-11-05 건국대학교 산학협력단 신규한 혐기성 세균용 비선택배지 조성물 및 그 제조방법
KR20140082964A (ko) * 2011-10-11 2014-07-03 아힘 바이오테라퓨틱스 에이비 혐기성 조건으로 배양된 인간 장내 미생물을 포함하는 조성물
KR20180041144A (ko) * 2015-08-25 2018-04-23 이뮨바이오테크 메디컬 스웨덴 아베 장 감염과 염증의 치료 및 예방을 위한 조성물 및 방법
KR20190004586A (ko) * 2017-07-04 2019-01-14 (주)바이오일레븐 장내미생물이식(fmt)을 위한 기증자 및 기증자 분변 선별방법

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR19990075377A (ko) * 1998-03-20 1999-10-15 유주현 신규한 호염성 단백질 분해효소 및 그를 생산하는 미생물
KR20120121100A (ko) * 2011-04-26 2012-11-05 건국대학교 산학협력단 신규한 혐기성 세균용 비선택배지 조성물 및 그 제조방법
KR20140082964A (ko) * 2011-10-11 2014-07-03 아힘 바이오테라퓨틱스 에이비 혐기성 조건으로 배양된 인간 장내 미생물을 포함하는 조성물
KR20180041144A (ko) * 2015-08-25 2018-04-23 이뮨바이오테크 메디컬 스웨덴 아베 장 감염과 염증의 치료 및 예방을 위한 조성물 및 방법
KR20190004586A (ko) * 2017-07-04 2019-01-14 (주)바이오일레븐 장내미생물이식(fmt)을 위한 기증자 및 기증자 분변 선별방법

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CHANG, S. E.: "Role of Intestinal Microbiota in Inflammatory Bowel Diseases", INTESTINAL RESEARCH., vol. 11, no. 3, 2013, pages 161 - 168 *
KIM, JUNG MOGG.: "Inflammatory Bowel Diseases and Enteric Microbiota", THE KOREAN JOURNAL OF GASTROENTEROLOGY, vol. 55, 2010, pages 4 - 18, XP055737026 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11680257B2 (en) 2015-05-11 2023-06-20 Mybiotics Pharma Ltd. Systems and methods for growing a biofilm of probiotic bacteria on solid particles for colonization of bacteria in the gut
US11679136B2 (en) 2016-05-25 2023-06-20 Mybiotics Pharma Ltd. Composition and methods for microbiota therapy

Similar Documents

Publication Publication Date Title
Pridgeon et al. Molecular identification and virulence of three Aeromonas hydrophila isolates cultured from infected channel catfish during a disease outbreak in west Alabama (USA) in 2009
Kitahara et al. Bacteroides plebeius sp. nov. and Bacteroides coprocola sp. nov., isolated from human faeces
Law et al. Streptomyces monashensis sp. nov., a novel mangrove soil actinobacterium from East Malaysia with antioxidative potential
WO2020075949A1 (fr) Nouvelle souche de lactobacillus plantarum atg-k2, atg-k6 ou atg-k8 et composition de prévention ou de traitement de la vaginite comprenant celle-ci
WO2014133322A1 (fr) Nouveau bactériophage et composition antibactérienne comprenant celui-ci
WO2014133323A1 (fr) Nouveau bactériophage et composition antibactérienne le comprenant
WO2019088379A1 (fr) Nouvelles bactéries lactiques et leur utilisation
WO2015088227A1 (fr) Nouvelles bactéries lactiques et composition contenant celles-ci pour la prévention ou le traitement de la diarrhée chez le nourrisson
Wang et al. Isolation of a Lysinibacillus fusiformis strain with tetrodotoxin-producing ability from puffer fish Fugu obscurus and the characterization of this strain
WO2020180037A1 (fr) Composition pour la culture d'un mimétique du microbiome intestinal humain, procédé de culture d'un mimétique du microbiome intestinal humain à l'aide d'un système de culture anaérobie, agent microbien préparé à l'aide de ce dernier, et son utilisation
EP2961836A1 (fr) Nouveau bactériophage et composition antibactérienne le comprenant
WO2023068434A1 (fr) Nouveau lactobacillus plantarum probio 93, et extrait et composition de celui-ci
Tsang et al. Gordonia hongkongensis sp. nov., isolated from blood culture and peritoneal dialysis effluent of patients in Hong Kong
WO2017179766A1 (fr) Méthode de préparation de fantômes de bactéries gram positif par traitement à l'acide chlorhydrique
WO2019004668A1 (fr) Nanovésicule dérivée de bactéries du genre proteus et son utilisation
US20230407241A1 (en) A novel probiotic streptococcus salivarius strain and its uses
KR20200106439A (ko) 인간 장내 미생물 군집 모사 배양용 조성물, 혐기 배양 체계를 이용한 인간 장내 미생물 군집의 모사 배양 방법, 이에 의해 제조된 미생물 제제 및 이의 용도
WO2022039561A1 (fr) Composition pour le traitement ou la prévention d'une infection par clostridium difficile
WO2019088421A1 (fr) Composition pour le traitement de lésions de cellules cutanées provoquées par les poussières fines, contenant des bactéries lactiques dérivées du thé vert
WO2021242056A1 (fr) Souche de l'espèce bifidobacterium et vésicule extracellulaire dérivée de cette dernière, et leurs utilisations anti-inflammatoires et antibactériennes
WO2025048505A1 (fr) Nouvelles bactéries lactiques et leur utilisation
WO2020197362A1 (fr) Microbe faecalibacterium sp. et composition anticancéreuse le comprenant
WO2024205297A1 (fr) Nouvelle souche de bifidobacterium et/ou de lactobacillus ou combinaison de celles-ci et leurs utilisations
WO2021029507A1 (fr) Souche de streptomyces racemochromogenes ayant une activité antimicrobienne contre pseudomonas syringae pv actinidiae et son utilisation
WO2024196187A1 (fr) Nouvelle souche latilactobacillus sakei cnsc001wb à fonction immunomodulatrice et son utilisation

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20766756

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 20766756

Country of ref document: EP

Kind code of ref document: A1