KR102392365B1 - Lactobacillus sakei RH1117 for improvement, prevention or treatment of rheumatoid arthritis and composition comprising the same - Google Patents
Lactobacillus sakei RH1117 for improvement, prevention or treatment of rheumatoid arthritis and composition comprising the same Download PDFInfo
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- KR102392365B1 KR102392365B1 KR1020200121728A KR20200121728A KR102392365B1 KR 102392365 B1 KR102392365 B1 KR 102392365B1 KR 1020200121728 A KR1020200121728 A KR 1020200121728A KR 20200121728 A KR20200121728 A KR 20200121728A KR 102392365 B1 KR102392365 B1 KR 102392365B1
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- lactobacillus sakei
- rheumatoid arthritis
- lactobacillus
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Abstract
본 발명은 신규한 락토바실러스 사케이(Lactobacillus sakei) RH1117 (수탁번호: KCCM12779P)에 관한 것으로, 락토바실러스 사케이(Lactobacillus sakei) RH1117 (수탁번호: KCCM12779P)를 포함하는 류마티스관절염 개선, 예방 또는 치료용 조성물을 제공할 수 있다.The present invention relates to a novel Lactobacillus sakei RH1117 (Accession No.: KCCM12779P), and Lactobacillus sakei RH1117 (Accession No.: KCCM12779P) for improving, preventing or treating rheumatoid arthritis containing RH1117 (Accession No.: KCCM12779P) compositions may be provided.
Description
본 발명은 신규한 락토바실러스 사케이(Lactobacillus sakei) RH1117 (수탁번호: KCCM12779P)에 관한 것으로, 더욱 상세하게는 락토바실러스 사케이(Lactobacillus sakei) RH1117 (수탁번호: KCCM12779P) 및 이를 포함하는 류마티스관절염 개선용, 치료용 또는 예방용 조성물에 관한 것이다.The present invention relates to a novel Lactobacillus sakei RH1117 (accession number: KCCM12779P), and more particularly, Lactobacillus sakei RH1117 (accession number: KCCM12779P) and rheumatoid arthritis improvement comprising the same It relates to a composition for use, treatment or prevention.
류마티스관절염(Rheumatoid arthritis)은 관절 활막의 지속적인 염증반응을 특징으로 하는 만성 염증성 전신질환으로, 골관절염에 의해 만성 관절염 중 두 번째로 많은 질병이다. 관절 활막의 지속적인 만성 염증반응으로 인해 관절의 연골 손상, 골 미란(bone erosion)이 일어나며, 결국은 관절의 파괴가 일어나 기능의 장애를 초래하는 것이 주요한 임상적 특징이다.Rheumatoid arthritis is a chronic inflammatory systemic disease characterized by a continuous inflammatory response of the joint synovial membrane, and is the second most common chronic arthritis disease caused by osteoarthritis. The main clinical features are that the joint cartilage damage and bone erosion occur due to the continuous chronic inflammatory reaction of the joint synovial membrane, and eventually the destruction of the joint results in functional impairment.
류마티스관절염은 대부분 호전과 악화를 반복하면서 진행하게 되며, 아직까지 예방하거나 완치를 시킬 수 있는 방법은 없지만 질환의 진행 속도를 늦추기 위해 주로 약제를 사용하게 된다. 기존에는 비스테로이드성 항염제와 스테로이드, 항류마티스약제와 TNF 차단제 등이 있으며, 비스테로이드성 항염제와 스테로이드는 염증에 의한 증상을 조절하여 질병의 증상을 완화시킬 수는 있지만 진행을 억제하지 못하는 것으로 알려져 있다. 게다가 상기의 약제들은 위장 장애, 장기간 복용으로 인한 부작용 등의 문제점이 있어 효과적인 치료법으로 사용하기에는 한계가 있다. 최근에는 이러한 한계를 극복하기 위해 관절염의 발생에 핵심적인 역할을 하는 사이토카인을 억제하는 기술을 개발하고 있다. Most rheumatoid arthritis progresses by repeating improvement and exacerbation, and although there is still no way to prevent or cure it, drugs are mainly used to slow the progression of the disease. Conventionally, there are nonsteroidal anti-inflammatory drugs and steroids, antirheumatic drugs and TNF blockers, etc. Nonsteroidal anti-inflammatory drugs and steroids can relieve symptoms of disease by controlling the symptoms of inflammation, but are not known to inhibit the progression. . In addition, the above drugs have problems such as gastrointestinal disorders and side effects due to long-term use, which limits their use as effective treatments. Recently, in order to overcome this limitation, a technology for inhibiting cytokines that play a key role in the development of arthritis is being developed.
선행 연구에 의하면, 대한민국 등록특허 제10-1355440호 (등록일자 2014.01.20)에는 항염증성 사이토카인인 IL-10의 발현을 증가시키면서 염증성 사이토카인인 IL-12 발현을 감소시킴으로써 류마티스관절염 예방 효능을 갖는 락토바실러스 헬베티쿠스(Lactobacillus helveticus) HY7801 및 이를 유효성분으로 함유하는 제품에 대해 개시된 바 있다. 이에 따라 기존 치료법의 한계점을 극복하고, 선행 연구의 균주에 비해 더 뛰어난 기능성을 가지면서 안전한 신규 치료제를 개발하는 것이 필요한 실정이다According to a previous study, Korean Patent Registration No. 10-1355440 (registration date: January 20, 2014) discloses rheumatoid arthritis prevention efficacy by decreasing the expression of IL-12, an inflammatory cytokine, while increasing the expression of IL-10, an anti-inflammatory cytokine. Having Lactobacillus helveticus ( Lactobacillus helveticus ) HY7801 and a product containing the same as an active ingredient has been disclosed. Accordingly, it is necessary to overcome the limitations of existing treatments and develop safe new treatments with superior functionality compared to the strains of previous studies.
본 발명은 류마티스관절염 개선, 예방 또는 치료 효과가 우수한 균주를 선별하고, 이를 포함한 조성물을 개발하여 제공하고자 한다.The present invention is to select a strain excellent in improving, preventing or treating rheumatoid arthritis, and to develop and provide a composition including the same.
본 발명은 류마티스관절염(Rheumatoid arthritis)의 예방 또는 치료능을 갖는 락토바실러스 사케이(Lactobacillus sakei) RH1117 (수탁번호: KCCM12779P)를 제공한다.The present invention provides a Lactobacillus sakei ( Lactobacillus sakei ) RH1117 (Accession No.: KCCM12779P) having a preventive or therapeutic ability of rheumatoid arthritis.
또한, 본 발명은 락토바실러스 사케이(Lactobacillus sakei) RH1117 (수탁번호: KCCM12779P)의 배양액, 상기 배양액의 농축물 및 상기 배양액의 건조 분말로 이루어지는 군에서 선택된 하나 이상을 포함하는 것을 특징으로 하는 류마티스관절염(Rheumatoid arthritis) 개선용 식품 조성물을 제공한다.In addition, the present invention is Lactobacillus sakei ( Lactobacillus sakei ) Rheumatoid arthritis characterized in that it comprises at least one selected from the group consisting of a culture solution of RH1117 (accession number: KCCM12779P), a concentrate of the culture solution, and a dry powder of the culture solution. (Rheumatoid arthritis) To provide a food composition for improving.
한편, 본 발명에 있어서, 상기 식품 조성물은, 바람직하게 유산균 발효유, 두유, 분유, 요구르트, 음료, 과립 또는 건강보조식품 중 선택되는 어느 하나인 것일 수 있다.Meanwhile, in the present invention, the food composition may be any one selected from lactic acid bacteria fermented milk, soy milk, powdered milk, yogurt, beverages, granules or health supplements.
또한, 본 발명은 락토바실러스 사케이(Lactobacillus sakei) RH1117 (수탁번호: KCCM12779P)의 배양액, 상기 배양액의 농축물 및 상기 배양액의 건조 분말로 이루어지는 군에서 선택된 하나 이상을 포함하는 것을 특징으로 하는 류마티스관절염(Rheumatoid arthritis) 예방 또는 치료용 약학 조성물을 제공한다.In addition, the present invention is Lactobacillus sakei ( Lactobacillus sakei ) Rheumatoid arthritis characterized in that it comprises at least one selected from the group consisting of a culture solution of RH1117 (accession number: KCCM12779P), a concentrate of the culture solution, and a dry powder of the culture solution. (Rheumatoid arthritis) To provide a pharmaceutical composition for preventing or treating.
본 발명은 선별한 락토바실러스 사케이(Lactobacillus sakei) RH1117 (수탁번호: KCCM12779P)를 포함하는 류마티스관절염 개선, 예방 또는 치료용 조성물을 제공할 수 있다.The present invention can provide a composition for improving, preventing or treating rheumatoid arthritis, including the selected Lactobacillus sakei RH1117 (Accession Number: KCCM12779P).
도 1은 마우스의 비장세포에서 본 발명의 락토바실러스 사케이(Lactobacillus sakei) RH1117에 의한 Th17 및 Treg의 발현과 IL-17 및 IL-10의 생성능에 대해 분석(유세포 분석, ELISA 분석)한 결과 그래프이다.
도 2는 정상인의 말초단핵구 세포에서 본 발명의 락토바실러스 사케이(Lactobacillus sakei) RH1117에 의한 Th17, Treg 및 B reg의 발현과 IL-17 및 IL-10의 생성능에 대해 분석(유세포 분석, ELISA 분석)한 결과 그래프이다.
도 3은 류마티스관절염 마우스 모델에서 본 발명의 락토바실러스 사케이(Lactobacillus sakei) RH1117에 의한 관절염 질환 활성에 대해 분석(A: 관절염 점수, 관절염 발병률, B: H&E 염색, 사프라닌 O 염색)한 결과 그래프이다.
도 4는 류마티스관절염 마우스 모델에서 본 발명의 락토바실러스 사케이(Lactobacillus sakei) RH1117에 의한 관절 조직에서의 조직화학 염색을 통해 염증성/병인 사이토카인의 발현을 분석한 결과 그래프이다.
도 5는 류마티스관절염 마우스 모델에서 본 발명의 락토바실러스 사케이(Lactobacillus sakei) RH1117에 의한 비장세포에서의 면역세포(Th17, Treg, Th1, Th2)의 아형 분석 결과 그래프이다.
도 6은 정상인의 말초단핵구 세포에서 본 발명의 락토바실러스 사케이(Lactobacillus sakei) RH1117에 의한 파골세포 분화능을 분석한 결과 그래프이다. 1 is a graph of the results of analysis (flow cytometry, ELISA analysis) for the expression of Th17 and Treg and the production ability of IL-17 and IL-10 by Lactobacillus sakei RH1117 of the present invention in splenocytes of mice. am.
2 is an analysis of the expression of Th17, Treg and B reg and the production ability of IL-17 and IL-10 by Lactobacillus sakei RH1117 of the present invention in peripheral mononuclear cells of a normal person (flow cytometry, ELISA analysis) ) is the result graph.
3 is a result of analysis (A: arthritis score, arthritis incidence rate, B: H&E staining, safranin O staining) for the arthritis disease activity caused by Lactobacillus sakei RH1117 of the present invention in a rheumatoid arthritis mouse model. It is a graph.
4 is a graph showing the results of analyzing the expression of inflammatory/pathogenic cytokines through histochemical staining in joint tissues with Lactobacillus sakei RH1117 of the present invention in a rheumatoid arthritis mouse model.
5 is a graph showing the results of subtype analysis of immune cells (Th17, Treg, Th1, Th2) in splenocytes by Lactobacillus sakei RH1117 of the present invention in a rheumatoid arthritis mouse model.
6 is a graph showing the result of analyzing osteoclast differentiation ability by Lactobacillus sakei RH1117 of the present invention in peripheral mononuclear cells of a normal person.
본 발명은 류마티스관절염(Rheumatoid arthritis)의 예방 또는 치료능을 갖는 락토바실러스 사케이(Lactobacillus sakei) RH1117 (수탁번호: KCCM12779P)를 제공한다.The present invention provides a Lactobacillus sakei ( Lactobacillus sakei ) RH1117 (Accession No.: KCCM12779P) having a preventive or therapeutic ability of rheumatoid arthritis.
본 발명에서는 건강한 인간 분변 샘플을 시료로 후벽균(Firmicutes) 문의 하위 속(genus)인 락토바실러스(Lactobacillus)를 분리하였다. 분리한 여러 종의 락토바실러스 중 류마티스관절염의 원인으로 지목되는 사이토카인 발현율 관련 in vitro 실험 결과가 가장 뛰어난 락토바실러스 사케이 (Lactobacillus sakei) RH1117 (수탁번호: KCCM12779P)를 선발하였고, 이는 류마티스관절염 개선, 예방 또는 치료에 대한 뛰어난 효능을 가지는 균주로 제공할 수 있다.In the present invention, a subgenus of the phylum Firmicutes , Lactobacillus , was isolated from a healthy human fecal sample as a sample. Among the isolated Lactobacillus species, Lactobacillus sakei has the best in vitro test results related to the expression rate of cytokines pointed out as the cause of rheumatoid arthritis RH1117 (Accession No.: KCCM12779P) was selected, which can be provided as a strain having excellent efficacy for improving, preventing or treating rheumatoid arthritis.
또한, 본 발명은 락토바실러스 사케이(Lactobacillus sakei) RH1117 (수탁번호: KCCM12779P)의 배양액, 상기 배양액의 농축물 및 상기 배양액의 건조 분말로 이루어지는 군에서 선택된 하나 이상을 포함하는 것을 특징으로 하는 류마티스관절염(Rheumatoid arthritis) 개선용 식품 조성물을 제공한다.In addition, the present invention is Lactobacillus sakei ( Lactobacillus sakei ) Rheumatoid arthritis characterized in that it comprises at least one selected from the group consisting of a culture solution of RH1117 (accession number: KCCM12779P), a concentrate of the culture solution, and a dry powder of the culture solution. (Rheumatoid arthritis) To provide a food composition for improving.
본 발명의 식품 조성물에 있어서, 상기 식품 조성물은, 일 예로, 육류, 곡류, 카페인 음료, 일반음료, 초콜릿, 빵류, 스낵류, 과자류, 사탕, 피자, 젤리, 면류, 껌류, 유제품류, 아이스크림류, 알코올성 음료, 술, 비타민 복합제 및 그 밖의 건강보조식품류중 선택되는 어느 하나 일수 있다. 더욱 바람직하게는 유산균 발효유, 두유, 분유, 요구르트, 음료, 과립 또는 건강보조식품류 중 선택되는 어느 하나인 것일 수 있으며 반드시 이에 한정되는 것은 아니다.In the food composition of the present invention, the food composition, for example, meat, grains, caffeinated beverages, general drinks, chocolate, breads, snacks, confectionery, candy, pizza, jelly, noodles, gums, dairy products, ice cream, It may be any one selected from alcoholic beverages, alcohol, vitamin complexes, and other health supplements. More preferably, it may be any one selected from lactic acid bacteria fermented milk, soy milk, powdered milk, yogurt, beverages, granules or health supplements, but is not necessarily limited thereto.
또한, 본 발명은 락토바실러스 사케이(Lactobacillus sakei) RH1117 (수탁번호: KCCM12779P)의 배양액, 상기 배양액의 농축물 및 상기 배양액의 건조 분말로 이루어지는 군에서 선택된 하나 이상을 포함하는 것을 특징으로 하는 류마티스관절염(Rheumatoid arthritis) 예방 또는 치료용 약학 조성물을 제공한다.In addition, the present invention is Lactobacillus sakei ( Lactobacillus sakei ) Rheumatoid arthritis characterized in that it comprises at least one selected from the group consisting of a culture solution of RH1117 (accession number: KCCM12779P), a concentrate of the culture solution, and a dry powder of the culture solution. (Rheumatoid arthritis) To provide a pharmaceutical composition for preventing or treating.
한편, 본 발명의 약학 조성물은 약제학적으로 허용 가능한 담체, 희석제 또는 부형제를 더욱 포함할 수 있다. 사용가능한 담체, 부형제 또는 희석제로는, 락토즈, 덱스트로즈, 수크로즈, 솔비톨, 만니톨, 자이리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로즈, 폴리비닐피롤리돈, 물, 메틸하이드록시벤조에이트, 프로필하이드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유가 있으며, 이중 선택되는 하나 이상을 사용할 수 있다. 또한, 치료 및 예방제가 약제인 경우 충진제, 항응집제, 윤활제, 습윤제, 향료, 유화제 또는 방부제 등이 추가적으로 포함될 수 있다.On the other hand, the pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, diluent or excipient. Usable carriers, excipients or diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, There are microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil, and at least one selected from among them may be used. In addition, when the therapeutic and prophylactic agent is a pharmaceutical, a filler, an anti-aggregant, a lubricant, a wetting agent, a fragrance, an emulsifier, or a preservative may be additionally included.
한편, 본 발명의 약학 조성물의 제형은 사용 방법에 따라 바람직한 형태로 제조될 수 있으며, 특히 포유동물에 투여된 후 활성 성분의 신속, 지속 또는 지연된 방출을 제공할 수 있도록 당업계에 공지된 방법을 채택하여 제형화 하는 것이 좋다. 구체적인 제형의 예로는 경고제(PLASTERS), 과립제(GRANULES), 로션제(LOTIONS), 리니멘트제(LINIMENTS), 리모나데제(LEMONADES), 방향수제(AROMATIC WATERS), 산제(POWDERS), 시럽제(SYRUPS), 안연고제(OPHTALMIC OINTMENTS), 액제(LIQUIDS AND SOLUTIONS), 에어로솔제(AEROSOLS), 엑스제(EXTRACTS), 엘릭실제(ELIXIRS), 연고제(OINTMENTS), 유동엑스제(FLUIDEXTRACTS), 유제(EMULSIONS), 현탁제(SUSPESIONS), 전제(DECOCTIONS), 침제(INFUSIONS), 점안제(OPHTHALMIC SOLUTIONS), 정제(TABLETS), 좌제(SUPPOSITIORIES), 주사제(INJECTIONS), 주정제(SPIRITS), 카타플라스마제(CATAPLSMA), 캅셀제(CAPSULES), 크림제(CREAMS), 트로키제(TROCHES), 틴크제(TINCTURES), 파스타제(PASTES), 환제(PILLS), 연질 또는 경질 젤라틴 캅셀 중 선택되는 어느 하나일 수 있다.On the other hand, the dosage form of the pharmaceutical composition of the present invention may be prepared in a desired form according to the method of use, and in particular, methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal It is better to adopt and formulate it. Examples of specific formulations include PLASTERS, GRANULES, LOTIONS, LINIMENTS, LEMONADES, AROMATIC WATERS, POWDERS, syrups ( SYRUPS, OPHTALMIC OINTMENTS, LIQUIDS AND SOLUTIONS, AEROSOLS, EXTRACTS, ELIXIRS, OINTMENTS, FLUIDEXTRACTS, EMULSIONS ), suspensions (SUSPESIONS), preparations (DECOCTIONS), infusions (INFUSIONS), eye drops (OPHTHALMIC SOLUTIONS), tablets (TABLETS), suppositories (SUPPOSITIORIES), injections (INJECTIONS), alcohol tablets (SPIRITS), CATAPLS ), capsules (CAPSULES), creams (CREAMS), troches (TROCHES), tinctures (TINCTURES), pastas (PASTES), pills (PILLS), it may be any one selected from soft or hard gelatin capsules.
한편, 본 발명의 약학조성물에 있어서, 투여량은 투여방법, 복용자의 연령, 성별 및 체중 및 질환의 중증도 등을 고려하여 결정하는 것이 좋다. 일 예로, 유효성분을 기준으로 하였을 때 1일 0.00001 내지 100 mg/kg (체중)으로 1회 이상 경구 투여 가능하다. 다만, 상기의 투여량은 예시하기 위한 일 예에 불과하며, 복용자의 상태와 의사의 처방에 의해 변화될 수 있다.On the other hand, in the pharmaceutical composition of the present invention, the dosage is preferably determined in consideration of the administration method, the age, sex and weight of the user, and the severity of the disease. As an example, based on the active ingredient, 0.00001 to 100 mg/kg (body weight) per day may be orally administered at least once. However, the above dosage is only an example for illustration, and may be changed by the condition of the user and the prescription of the doctor.
한편, 본 발명의 균주는 하기 실험에 의할 경우, 염증성 사이토카인인 IL-17을 감소시키고, 항염증성 사이토카인인 IL-10을 증가시킴으로써 류마티스관절염에 영향을 주는 사이토카인의 발현을 조절함으로써 류마티스관절염 질환에 대한 제어 효과를 발휘한다. 또한, 본 발명의 균주는 염증 정도, 뼈 파괴 및 연골 파괴 정도를 감소시키고, 파골세포의 분화를 억제시킴으로써 류마티스관절염 관절 조직의 파괴에 대한 제어 효과를 발휘한다. 이를 통해 본 발명에서는 안전하면서도 류마티스관절염의 개선, 예방 또는 치료에 효과적인 균주를 제공할 수 있고, 이를 포함한 식품 및 약학 조성물을 제공할 수 있다.On the other hand, according to the following experiment, the strain of the present invention reduces the inflammatory cytokine IL-17 and increases the anti-inflammatory cytokine IL-10 by regulating the expression of cytokines affecting rheumatoid arthritis. It exerts a controlling effect on arthritic diseases. In addition, the strain of the present invention exerts a controlling effect on the destruction of rheumatoid arthritis joint tissue by reducing the degree of inflammation, bone destruction and cartilage destruction, and suppressing the differentiation of osteoclasts. Through this, in the present invention, it is possible to provide a safe and effective strain for the improvement, prevention or treatment of rheumatoid arthritis, and food and pharmaceutical compositions including the same can be provided.
이하, 본 발명의 내용을 하기 실시예를 통해 더욱 상세히 설명하고자 한다. 다만, 본 발명의 권리범위가 하기 실시예에만 한정되는 것은 아니고, 그와 등가의 기술적 사상의 변형까지를 포함한다.Hereinafter, the content of the present invention will be described in more detail through the following examples. However, the scope of the present invention is not limited only to the following examples, and includes modifications of technical ideas equivalent thereto.
[실시예 1: 락토바실러스 사케이([Example 1: Lactobacillus Sake ( Lactobacillus sakeiLactobacillus sakei ) RH1117의 선발]) Selection of RH1117]
동정된 락토바실러스 사케이는 건강인의 대변에서 100 mg의 시료를 취하여 단일 콜로니(colony)를 얻기 위해 멸균 인산완충식염수(PBS, pH 7.4)으로 십진 희석하여 4가지 고체배지에 획선도말하였다. 0.05% L-cysteine-HCl(cys)을 함유한 de Man-Rogosa-Sharpe (MRS) (BD DifcoTM, Franklin Lakes, NJ, USA) 고체배지, Lactobacillus selection Agar (LBS) (BD DifcoTM, Franklin Lakes, NJ, USA) 고체배지, Transgalacto-oligosaccharide (TOS) antibiotic mupirocin 함유 (TOS-MUP) (MB cell, MB-T0892, 서울, 한국) 고체배지, Columbia (MB cell, MB-C0700, 서울, 한국) 고체배지에 도말하여 37℃, 30시간 혐기적 조건으로 배양하였다. 각 배지에 형성된 단일 콜로니 10개를 임의 선택하여 cMRS 액체배지에서 37℃, 30시간 혐기적 조건으로 정치 배양하였다. 각 균의 동정은 16S rRNA 시퀀싱 분석을 Macrogen(주)(서울, 한국)에 의뢰하여 실시하였다. 이후 동정이 완료된 균은 50% 멸균 Glycerol solution에 50% 배양액을 넣고 -70℃에서 보관하며 사용하였다.The identified Lactobacillus saccharis was diluted decimal with sterile phosphate-buffered saline (PBS, pH 7.4) to obtain a single colony by taking a sample of 100 mg from the feces of a healthy person, and smearing it on four solid media. de Man-Rogosa-Sharpe (MRS) (BD Difco TM , Franklin Lakes, NJ, USA) solid medium containing 0.05% L-cysteine-HCl (cys), Lactobacillus selection Agar (LBS) (BD Difco TM , Franklin Lakes) , NJ, USA) Solid medium, Transgalacto-oligosaccharide (TOS) containing antibiotic mupirocin (TOS-MUP) (MB cell, MB-T0892, Seoul, Korea) Solid medium, Columbia (MB cell, MB-C0700, Seoul, Korea) It was spread on a solid medium and incubated anaerobically at 37°C for 30 hours. 10 single colonies formed in each medium were randomly selected and cultured in cMRS liquid medium at 37° C. under anaerobic conditions for 30 hours. The identification of each bacteria was performed by requesting 16S rRNA sequencing analysis to Macrogen (Seoul, Korea). After the identification was completed, 50% culture solution was added to 50% sterile glycerol solution and stored at -70°C before use.
이와 같은 과정을 통해 후벽균(Firmicutes) 문의 하위 속(genus)인 락토바실러스(Lactobacillus) 중 류마티스관절염의 원인으로 지목되는 사이토카인 발현율 관련 in vitro 실험결과가 가장 뛰어난 락토바실러스 사케이 (Lactobacillus sakei) 1117를 선발하였고, 이는 류마티스관절염 개선, 예방 또는 치료에 대한 뛰어난 효능을 가지는 균주로 제공할 수 있다. 선발에 대한 내용은 아래와 같다.Through this process, Lactobacillus sakei 1117 has the best in vitro experimental results related to the cytokine expression rate pointed out as the cause of rheumatoid arthritis among Lactobacillus , a subgenus of the Firmicutes phylum . was selected, which can be provided as a strain having excellent efficacy for improving, preventing or treating rheumatoid arthritis. Details of the selection are as follows.
본 발명의 락토바실러스 사케이는 다양한 균종 중에서 면역 조절 기능이 우수한 것으로 확인되어 선발된 균종이다. 선발한 방법은 인간의 말초단핵구 세포에 antiCD3 자극에 따른 T 세포 활성화 조건에 각각의 균들을 처리하여 염증 및 류마티스관절염 병인 사이토카인으로 알려져 있는 IL-17을 가장 많이 억제시키는 균과 항염증사이토카인인 IL-10을 억제시키지 않거나 유지시키는 균 중에서 그 기능이 우수한 것을 선발하였다. 그러한 결과를 토대로 류마티스관절염 동물모델에서의 치료 효과를 확인하게 되었다.Lactobacillus saccharis of the present invention is a strain selected from among various strains confirmed to have excellent immunomodulatory function. The selected method treats human peripheral mononuclear cells under T cell activation conditions according to antiCD3 stimulation, thereby inhibiting most of IL-17, known as a cytokine that causes inflammation and rheumatoid arthritis, and an anti-inflammatory cytokine. Among the bacteria that do not inhibit or maintain IL-10, those having excellent functions were selected. Based on such results, the therapeutic effect in the rheumatoid arthritis animal model was confirmed.
본 발명에서 선발한 락토바실러스 사케이(Lactobacillus sakei) RH1117를 한국생명공학연구원에 2020년 8월 20일자로 기탁한 후, 수탁번호: KCCM12779P를 부여받았다.After the Lactobacillus sakei RH1117 selected in the present invention was deposited at the Korea Research Institute of Bioscience and Biotechnology on August 20, 2020, it was given an accession number: KCCM12779P.
하기에서는 선발한 본 발명의 락토바실러스 사케이(Lactobacillus sakei) RH1117 (수탁번호: KCCM12779P)를 어떤 효능이 나타나는지를 확인하고자 동물 실험을 수행하였다.In the following, an animal experiment was performed to confirm the efficacy of the selected Lactobacillus sakei RH1117 (accession number: KCCM12779P) of the present invention.
[실험예 1 : 마우스 및 인간 세포에서의 락토바실러스 사케이([Experimental Example 1: Lactobacillus saccharis in mouse and human cells ( Lactobacillus sakeiLactobacillus sakei ) RH1117에 의한 류마티스관절염 관련 인자 발현 테스트]) rheumatoid arthritis-related factor expression test by RH1117]
본 실험예에서는 마우스의 비장세포 및 인간 말초단핵구 세포에서 상기 실시예 1의 락토바실러스 사케이(Lactobacillus sakei) RH1117에 의한 류마티스관절염 관련 인자 발현 양상을 확인하고자 하였다.In this experimental example, in mouse splenocytes and human peripheral mononuclear cells, the The purpose of this study was to confirm the expression pattern of rheumatoid arthritis-related factors by Lactobacillus sakei RH1117.
1) 마우스 비장세포에서 Th17, IL-17 억제 및 IL-10 증가 효과1) Th17, IL-17 inhibition and IL-10 increase effect in mouse splenocytes
정상 마우스의 비장세포에 T세포의 활성 조건을 유도하기 위해 antiCD3 0.5 ug/ml로 자극한 후, 락토바실러스 사케이(Lactobacillus sakei) RH1117를 1 ug/ml처리하여 3일간 배양하였다. 비교를 위해 락토바실러스 사케이 RH1117 미투여군(vehicle, -로 표시)과 양성대조군 Nil을 사용하였다.After stimulation with antiCD3 0.5 ug/ml to induce T-cell activation conditions in normal mouse splenocytes, Lactobacillus sakei RH1117 was treated with 1 ug/ml and cultured for 3 days. For comparison, the Lactobacillus Sakei RH1117 untreated group (vehicle, indicated by -) and the positive control group Nil were used.
3일간 배양한 Th17(CD4+ IL-17+) 세포의 발현을 유세포 분석한 결과, 도 1의 A와 같이 락토바실러스 사케이(Lactobacillus sakei) RH1117를 처리할 시, vehicle에 비해 Th17 세포가 유의적으로 감소하였다. 상기와 같은 조건으로 3일간 배양한 supernatant를 ELISA 분석한 결과, 도 1의 B와 같이 락토바실러스 사케이(Lactobacillus sakei) RH1117를 처리할 시, vehicle에 비해 염증성 사이토카인인 IL-17이 유의미하게 감소하였다. As a result of flow cytometry analysis of the expression of Th17 (CD4+ IL-17+) cells cultured for 3 days, as shown in FIG. 1A, when Lactobacillus sakei RH1117 was treated, Th17 cells were significantly increased compared to vehicle. decreased. As a result of ELISA analysis of supernatants cultured for 3 days under the same conditions as above, as shown in FIG. 1B, when Lactobacillus sakei RH1117 was treated, IL-17, an inflammatory cytokine, was significantly reduced compared to vehicle. did
또한, 상기와 같은 조건으로 3일간 배양한 Treg(CD4+ CD25high Foxp3+) 세포의 발현을 유세포 분석한 결과, 도 1의 C와 같이 락토바실러스 사케이(Lactobacillus sakei) RH1117를 처리할 시, vehicle에 비해 Treg 세포가 감소하였다. 상기와 같은 조건으로 3일간 배양한 supernatant를 ELISA 분석한 결과, 도 1의 D와 같이 락토바실러스 사케이(Lactobacillus sakei) RH1117를 처리할 시, vehicle에 비해 항염증성 사이토카인인 IL-10이 유의미하게 증가하였다. In addition, as a result of flow cytometry analysis of the expression of Treg (CD4+ CD25high Foxp3 + ) cells cultured for 3 days under the same conditions as above, as shown in C of FIG. 1 , when treated with Lactobacillus sakei RH1117, compared to vehicle Treg cells were reduced. As a result of ELISA analysis of supernatants cultured for 3 days under the same conditions as above, as shown in D of FIG. 1, when Lactobacillus sakei RH1117 was treated, IL-10, an anti-inflammatory cytokine, was significantly higher than that of vehicle. increased.
2) 인간 말초단핵구 세포에서 Th17, IL-17 억제 및 IL-10, B reg 세포 증가 효과2) Effect of inhibiting Th17 and IL-17 and increasing IL-10 and B reg cells in human peripheral mononuclear cells
정상인의 말초단핵구 세포에서 T세포의 활성 조건을 유도하기 위해 antiCD3 0.5 ug/ml로 자극한 후, 락토바실러스 사케이(Lactobacillus sakei) RH1117를 0.1, 1 ug/ml 처리하여 3일간 배양하였다. 비교를 위해 락토바실러스 사케이 RH1117 미투여군(vehicle, -로 표시)과 양성대조군 Nil을 사용하였다.After stimulation with antiCD3 0.5 ug/ml to induce T-cell activation conditions in normal human peripheral mononuclear cells, Lactobacillus sakei RH1117 was treated with 0.1, 1 ug/ml, and cultured for 3 days. For comparison, the Lactobacillus Sakei RH1117 untreated group (vehicle, indicated by -) and the positive control group Nil were used.
3일간 배양한 Th17(CD4+ IL-17+) 세포의 발현을 유세포 분석한 결과, 도 2의 A와 같이 락토바실러스 사케이(Lactobacillus sakei) RH1117를 처리할 시, 처리농도에 따라 Th17 세포가 유의적으로 감소하였다. 또한, 상기와 같은 조건으로 3일간 배양한 Treg(CD4+ CD25high Foxp3+)의 발현을 유세포 분석한 결과, 락토바실러스 사케이(Lactobacillus sakei) RH1117를 처리할 시, Treg가 유지되었다. 또한, 상기와 같은 조건으로 3일간 배양후 supernatant를 ELISA 분석한 결과, 도 2의 B와 같이 락토바실러스 사케이(Lactobacillus sakei) RH1117를 처리할 시, 처리 농도에 따라 vehicle에 비해 염증성 사이토카인인 IL-17이 유의미하게 감소하고, 항염증성 사이토카인인 IL-10은 유의미하게 증가하였다. As a result of flow cytometry analysis of the expression of Th17 (CD4+ IL-17+) cells cultured for 3 days, as shown in FIG. 2A, when RH1117 was treated, Th17 cells were significantly increased according to the treatment concentration. decreased to In addition, as a result of flow cytometry analysis of the expression of Treg (CD4+ CD25high Foxp3 + ) cultured for 3 days under the same conditions as above, when treated with Lactobacillus sakei RH1117, Treg was maintained. In addition, as a result of ELISA analysis of supernatants after culturing for 3 days under the same conditions as above, as shown in FIG. 2B, when Lactobacillus sakei RH1117 was treated, IL, an inflammatory cytokine, compared to vehicle, depending on the treatment concentration -17 was significantly decreased, and IL-10, an anti-inflammatory cytokine, was significantly increased.
한편, 정상인의 말초단핵구 세포에서 B세포의 활성 조건을 유도하기 위해 LPS 100 ng/ml로 자극한 후, 락토바실러스 사케이(Lactobacillus sakei) RH1117를 0.1, 1, 10 ug/ml 처리하여 3일간 배양하였다. 3일간 배양한 B reg(CD19+ IL-10+)세포의 발현을 유세포 분석한 결과, 도 2의 C와 같이 락토바실러스 사케이(Lactobacillus sakei) RH1117를 처리할 시, 처리농도에 따라 CD19+ IL-10+세포가 농도 의존적으로 증가하였음을 확인할 수 있었다. 이상 종합하면, IL-10이 증가되는 현상은 Treg(CD4+ CD25high Foxp3+)은 증가 경향이 없고 유지되었으나 세포를 배양한 sup에서는 IL-10이 유의적, 농도 의존적으로 증가됨을 관찰하였고, 이러한 결과는 B reg(CD19+ IL-10+)인 세포의 증가와 관련이 있음을 알 수 있었다. 즉, 락토바실러스 사케이(Lactobacillus sakei) RH1117에 의해 IL-10이 증가되는 현상은 Treg 유래가 아닌 B reg의 증가와 영향이 있는 것을 알 수 있었다.Meanwhile, in normal human peripheral mononuclear cells, after stimulation with
[실험예 2 : 류마티스관절염 전임상 동물모델에서의 락토바실러스 사케이([Experimental Example 2: Lactobacillus saccharis in rheumatoid arthritis preclinical animal model ( Lactobacillus sakeiLactobacillus sakei ) RH1117에 의한 류마티스관절염 질환 관련 효능 테스트]) Efficacy test related to rheumatoid arthritis disease by RH1117]
본 실험예에서는 류마티스관절염 전임상 동물모델에서 상기 실시예 1의 락토바실러스 사케이(Lactobacillus sakei) RH1117에 의한 류마티스관절염 질환 관련 효능 테스트를 하였다.In this experimental example, in the preclinical animal model of rheumatoid arthritis, the Lactobacillus sakei ( Lactobacillus sakei ) Efficacy related to rheumatoid arthritis disease by RH1117 was tested.
1) 류마티스관절염 전임상 동물모델에서 락토바실러스 사케이(1) In a preclinical animal model of rheumatoid arthritis, Lactobacillus saccharis ( Lactobacillus sakeiLactobacillus sakei ) RH1117에 의한 관절염 질환 제어 효능) Efficacy of arthritis disease control by RH1117
관절염 질환에 대한 제어 효능을 평가하기 위해 정상인 유래의 락토바실러스 사케이(Lactobacillus sakei) RH1117와 관절염 환자 유래의 락토바실러스 사케이(Lactobacillus sakei) RH1114를 각각 사용하였다.In order to evaluate the control efficacy on arthritis disease, Lactobacillus sakei RH1117 derived from normal people and Lactobacillus sakei RH1114 derived from arthritis patients were used, respectively.
DBJ1/J 마우스에 type Ⅱ 콜라겐(collagen)(CⅡ)과 CFA(adjuvant)를 1:1로 혼합하여 마우스당 100 ㎍의 CⅡ를 tail base에 50 ㎕ 용량으로 주사하였다. 2주 후, CⅡ와 IFA 1:1로 혼합한 혼합액을 100 ㎍/50 ㎕로 2차 주사하는 방법으로 관절염 동물을 제작하였다. 관절염 발병 3주 후부터 관절염 환자 유래의 락토바실러스 사케이(Lactobacillus sakei) RH1114, 정상인 유래의 락토바실러스 사케이(Lactobacillus sakei) RH1117를 50 mg/kg씩 매일 경구투여하였다. 대조군으로는 락토바실러스 사케이 RH1117 미투여군(vehicle, -로 표시)과, 항 류마티스약제이면서 현재 임상에서 사용중인 메토트렉세이트 투여군(MTX)을 두었다. 메토트렉세이트는 5 mg/kg씩 매일 경구투여하였다. 관절염 유발 후 7주차에 마우스를 희생하여 관절을 얻어서 파라핀 블록을 제작하였다. DBJ1/J mice were mixed with type II collagen (CII) and CFA (adjuvant) at a 1:1 ratio, and 100 μg of CII per mouse was injected into the tail base at a volume of 50 μl. Two weeks later, arthritic animals were prepared by a second injection of 100 μg/50 μl of a mixed solution of CII and IFA 1:1. From 3 weeks after the onset of arthritis, 50 mg/kg of Lactobacillus sakei RH1114 derived from arthritis patients and Lactobacillus sakei RH1117 derived from normal persons were orally administered daily. As a control group, the Lactobacillus Sakei RH1117 untreated group (vehicle, indicated by -) and the antirheumatic drug methotrexate administered group (MTX) currently used in clinical practice were included. Methotrexate was orally administered at 5 mg/kg daily. At 7 weeks after induction of arthritis, mice were sacrificed to obtain joints to prepare paraffin blocks.
그 결과, 도 3의 A(왼쪽: 관절염 증상점수(Arthritis score), 오른쪽: 관절염 발병률(Incidence))와 같이 락토바실러스 사케이(Lactobacillus sakei)를 처리함으로써, 관절염 질환에 관한 제어 효능을 확인할 수 있었다. 또한, 도 3의 B와 같이 H&E 염색/사프라닌 O 염색 분석을 통해 관절조직에서의 염증 점수(inflammation score), 뼈 손상(Bone damage), 연골 손상(Cartilage damage) 정도를 확인한 결과, 락토바실러스 사케이(Lactobacillus sakei)를 처리하지 않은 vehicle에 비해 락토바실러스 사케이(Lactobacillus sakei)를 처리할 때, 염증 점수, 뼈 손상, 연골 손상 정도에서 유의하게 감소하였고 이를 통해 염증세포의 침투(infiltration)와 연골 보호 효능이 나타나는 것을 확인할 수 있었다. 정상인 유래의 락토바실러스 사케이(Lactobacillus sakei) RH1117과 환자유래의 락토바실러스 사케이(Lactobacillus sakei) RH1114를 이용한 관절염 지수는 정상인 유래 RH1117과 환자유래 RH1114가 유사하였으나 발병율에서는 정상인 유래 RH1117이 임상에서 사용중인 메토트렉세이트(Methotrexate)보다 더 우위의 효과를 보였다. 또한, 조직학적 분석을 통해 확인해보면 환자유래 RH1114보다 정상인 유래 RH1117이 연골 손상(Cartilage damage) 부분에서는 더 효과가 좋은 것이 관찰되었다.As a result, by treating Lactobacillus sakei as shown in FIG. 3A (Left: Arthritis score, Right: Arthritis Incidence), it was possible to confirm the control efficacy on arthritis disease. . In addition, as shown in FIG. 3B, as a result of confirming the degree of inflammation score, bone damage, and cartilage damage in joint tissues through H&E staining/safranin O staining analysis, Lactobacillus Compared to the vehicle not treated with sake ( Lactobacillus sakei ), when Lactobacillus sakei was treated, the inflammation score, bone damage, and cartilage damage were significantly reduced, and through this, the infiltration of inflammatory cells and It was confirmed that the cartilage protection effect appears. The arthritis index using Lactobacillus sakei RH1117 derived from a normal person and Lactobacillus sakei RH1114 from a patient was similar to RH1117 from a normal person and RH1114 from a patient. It was more effective than methotrexate. In addition, when confirmed through histological analysis, it was observed that normal human-derived RH1117 was more effective in cartilage damage than patient-derived RH1114.
2) 류마티스관절염 전임상 동물모델에서 락토바실러스 사케이(2) In a preclinical animal model of rheumatoid arthritis, Lactobacillus saccharis ( Lactobacillus sakeiLactobacillus sakei ) RH1117에 의한 염증성/병인 사이토카인 억제 효능) Inhibitory efficacy of inflammatory/pathogenic cytokines by RH1117
상기와 같이 제조한 관절염 모델에서, 정상인 유래의 락토바실러스 사케이(Lactobacillus sakei) RH1117와 관절염 환자 유래의 락토바실러스 사케이(Lactobacillus sakei) RH1114를 투여한 관절 조직의 조직화학 염색 분석을 통해 염증성/병인 사이토카인의 발현을 분석하였다. 그 결과, 도 4와 같이 락토바실러스 사케이(Lactobacillus sakei)를 처리하지 않은 vehicle에 비해 락토바실러스 사케이(Lactobacillus sakei)를 처리할 때, 염증성 사이토카인인 TNF-α, IL-1β, IL-6과 병인 사이토카인인 IL-17이 효과적으로 억제되는 것을 확인할 수 있었다. 특히 환자유래의 락토바실러스 사케이(Lactobacillus sakei) RH1114보다 정상인 유래의 락토바실러스 사케이(Lactobacillus sakei) RH1117가 염증성 사이토카인인 IL-1β와 류마티스관절염의 주요 병인 사이토카인인 IL-17을 억제시키는 효과가 뚜렷하게 차이남을 확인할 수 있었다.In the arthritis model prepared as described above, through histochemical staining analysis of joint tissues administered with Lactobacillus sakei RH1117 derived from a normal person and Lactobacillus sakei RH1114 derived from an arthritis patient, inflammatory/pathogenesis The expression of cytokines was analyzed. As a result, when treated with Lactobacillus sakei compared to the vehicle not treated with Lactobacillus sakei as shown in FIG. 4, inflammatory cytokines TNF-α, IL-1β, IL-6 It was confirmed that IL-17, a cytokine that is a pathogenic agent, was effectively inhibited. In particular, the effect of Lactobacillus sakei RH1117 derived from normal human beings to inhibit IL-1β, an inflammatory cytokine, and IL-17, a cytokine that is a major cause of rheumatoid arthritis, rather than patient-derived Lactobacillus sakei RH1114. was found to be distinctly different.
3) 류마티스관절염 전임상 동물모델에서 락토바실러스 사케이(3) In a preclinical animal model of rheumatoid arthritis, Lactobacillus saccharis ( Lactobacillus sakeiLactobacillus sakei ) RH1117에 의한 면역 세포 아형 분석 테스트) Immune cell subtyping test by RH1117
상기와 같이 제조한 관절염 모델에서, 실험종료 시점에 비장세포를 얻어서 면역세포(Th17, Treg, Th1, Th2)의 아형 분석을 수행하였다. 그 결과, 도 5와 같이 락토바실러스 사케이(Lactobacillus sakei) RH1117를 처리할 시, vehicle에 비해 Th17(CD4+ IL-17+) 세포가 효과적으로 억제되었다. 한편, 같은 조건으로 수행하였을 때, Treg(CD4+ CD25high Foxp3+), Th1(CD4+ IFN-r+), Th2(CD4+ IL-4+)에서는 유의미한 결과가 관찰되지 않았다.In the arthritis model prepared as described above, splenocytes were obtained at the end of the experiment, and subtype analysis of immune cells (Th17, Treg, Th1, Th2) was performed. As a result, as shown in FIG. 5, when Lactobacillus sakei RH1117 was treated, Th17 (CD4+ IL-17+) cells were effectively inhibited compared to vehicle. On the other hand, when performed under the same conditions, no significant results were observed in Treg (CD4+ CD25high Foxp3 + ), Th1 (CD4+ IFN-r+), and Th2 (CD4+ IL-4+).
[실험예 3 : 인간 세포에서의 락토바실러스 사케이([Experimental Example 3: Lactobacillus saccharis in human cells ( Lactobacillus sakeiLactobacillus sakei ) RH1117에 의한 파골세포 분화 억제 효능]) Efficacy of inhibiting osteoclast differentiation by RH1117]
본 실험예에서는 인간 말초단핵구 세포에서 상기 실시예 1의 락토바실러스 사케이(Lactobacillus sakei) RH1117에 의한 파골세포 분화 억제 효능을 확인하고자 하였다.In this experimental example, in human peripheral mononuclear cells, the The purpose of this study was to determine the efficacy of inhibiting osteoclast differentiation by Lactobacillus sakei RH1117.
정상인의 말초단핵구 세포에서 MCSF (100 ng/ml)+ RANKL(30 ng/ml)을 처리하여 파골세포 분화를 유도한 후, 락토바실러스 사케이(Lactobacillus sakei) RH1117를 0.1, 1, 10 ug/ml 처리하여 72시간 배양하였다. 파골세포의 분화 정도를 관찰하였다. 관찰을 위해 TRAP+ (tartrate resistant acid phosphatase) 염색을 수행하여 파골세포의 개수를 카운트하여 분화 억제능을 조사하였다.After inducing osteoclast differentiation by treatment with MCSF (100 ng/ml)+RANKL (30 ng/ml) in normal human peripheral mononuclear cells, Lactobacillus sakei RH1117 was added at 0.1, 1, 10 ug/ml treated and cultured for 72 hours. The degree of differentiation of osteoclasts was observed. For observation, TRAP+ (tartrate resistant acid phosphatase) staining was performed to count the number of osteoclasts, and the differentiation inhibitory ability was investigated.
그 결과, 도 6의 A와 같이 락토바실러스 사케이(Lactobacillus sakei) RH1117를 1, 10 ug/ml 처리할 시, vehicle에 비해 TRAP + 세포가 유의하게 감소되었다. 또한, 파골세포로부터 RNA를 얻어 Real time PCR을 수행하여 파골세포 분화 마커(TRAP, Calcitoin receptor, Cathepsin K)를 분석한 결과, 락토바실러스 사케이(Lactobacillus sakei) RH1117를 1, 10 ug/ml 처리할 시, vehicle에 비해 현저하게 감소되었다. 상기 결과를 통해 본 발명의 락토바실러스 사케이(Lactobacillus sakei) RH1117가 파골세포 분화 억제능이 있음을 알 수 있었다. As a result, as shown in FIG. 6A, when Lactobacillus sakei RH1117 was treated with 1, 10 ug/ml, TRAP + cells were significantly reduced compared to vehicle. In addition, as a result of obtaining RNA from osteoclasts and performing real-time PCR to analyze osteoclast differentiation markers (TRAP, Calcitoin receptor, Cathepsin K), Lactobacillus sakei RH1117 was treated with 1, 10 ug/ml City, it was significantly reduced compared to the vehicle. Through the above results, it can be seen that the Lactobacillus sakei RH1117 of the present invention has the ability to inhibit osteoclast differentiation.
기탁기관명 : 한국미생물보존센터(국외)Name of deposit institution: Korea Microorganism Conservation Center (Overseas)
수탁번호 : KCCM12779PAccession number: KCCM12779P
수탁일자 : 2020820Deposit date: 2020820
Claims (4)
Interleukin-17 (IL-17) secretion inhibitory effect, interleukin-10 (IL-10) secretion increase effect according to the increase of regulatory B cells (B reg cells) and osteoclast differentiation inhibitory ability, rheumatoid arthritis (Rheumatoid arthritis) ) Lactobacillus sakei having a preventive or therapeutic ability ( Lactobacillus sakei ) RH1117 (Accession No.: KCCM12779P).
Interleukin-17 (IL-17) secretion inhibitory effect, interleukin-10 (IL-10) secretion increase effect according to the increase of regulatory B cells (B reg cells) and osteoclast differentiation inhibitory ability, rheumatoid arthritis (Rheumatoid arthritis) ) of Lactobacillus sakei ( Lactobacillus sakei ) RH1117 (Accession No.: KCCM12779P) having a preventive or therapeutic ability, a concentrate of the culture medium, and a dry powder of the culture medium comprising at least one selected from the group consisting of A food composition for improving rheumatoid arthritis.
상기 식품 조성물은,
유산균 발효유, 두유, 분유, 요구르트, 과립 또는 건강보조식품 중 선택되는 어느 하나인 것을 특징으로 하는 식품 조성물.
3. The method of claim 2,
The food composition,
Food composition, characterized in that any one selected from lactic acid bacteria fermented milk, soy milk, powdered milk, yogurt, granules or health supplements.
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| KR101761379B1 (en) * | 2014-06-23 | 2017-08-04 | 한국식품연구원 | Novel Lactobacillus sakei K040706 for multi function and culture method thereof |
| KR20160108645A (en) * | 2015-03-04 | 2016-09-20 | 을지대학교 산학협력단 | Pharmaceutical and nutraceutical anti-inflammatory composition comprising supernatant solution of fermented soymilk |
| KR102242668B1 (en) * | 2019-02-26 | 2021-04-21 | 전남대학교산학협력단 | Composition for improving, preventing or treating bone diseases comprising Lactobacillus sakei CVL-001 or culture medium thereof |
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2020
- 2020-09-21 KR KR1020200121728A patent/KR102392365B1/en active IP Right Grant
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009269906A (en) * | 2008-04-30 | 2009-11-19 | Genmont Biotech Inc | Lactobacillus isolated strain having anti-inflammatory activity and use thereof |
| KR102123505B1 (en) | 2019-03-26 | 2020-06-17 | 한국식품연구원 | Lactobacillus sakei WIKIM0109 having anti-arthritis activity and composition for comprising the same |
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| KR20220039911A (en) | 2022-03-30 |
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