KR102040117B1 - Bifidobacterium bifidum MG731 and composition for preventing or treating cancers comprising the same - Google Patents
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Abstract
Description
본 발명은 암을 예방 또는 치료하는 우수한 효과를 갖는 신규한 비피도박테리움 비피덤 MG731 (Bifidobacterium bifidum MG731) 균주에 관한 것이다. The present invention relates to a novel Bifidobacterium bifidum MG731 (Bifidobacterium bifidum MG731) strain having an excellent effect of preventing or treating cancer.
구체적으로, 본 발명의 신규한 비피도박테리움 비피덤 MG731 균주는 암세포의 증식을 억제하는 효과를 가질 뿐만 아니라, 암세포의 이동성을 저하시키고 신혈관의 생성에 관여하는 유전자의 발현을 조절하는 효과를 동시에 가짐으로써, 기존의 비피도박테리움 비피덤 균주에 비하여 월등하게 우수한 효과를 갖는다. 뿐만 아니라, 본 발명의 비피도박테리움 비피덤 MG731 균주는 항염증, 항산화 및 면역증강 효과를 가진다.Specifically, the novel Bifidobacterium bifidum MG731 strain of the present invention not only has the effect of inhibiting the proliferation of cancer cells, but also has the effect of reducing the mobility of cancer cells and regulating the expression of genes involved in the generation of new blood vessels. By having it at the same time, it has a superior effect compared to the existing Bifidobacterium bifidum strain. In addition, the Bifidobacterium bifidum MG731 strain of the present invention has anti-inflammatory, antioxidant and immune enhancing effects.
인체의 장은 다양한 종류의 세균들의 공생으로 음식물의 소화에 따른 영양소 흡수 및 체내에 불필요한 물질을 체외로 배설하는 중요한 기능을 가진다. 또한 장내 세균에 의한 환경의 변화는 장 면역과도 직결되고 이는 결국 체내 면역력에도 영향을 미친다. 이러한 장내 건강에 유익한 미생물들에 대한 연구가 수년간 지속되어 오면서 유산균의 장 기능, 면역력 및 체내 대사활동 관여에 대한 연구결과들이 전세계적으로 활발히 발표되고 있다. 살아있는 미생물체를 일컫는 프로바이오틱스 (probiotics)는 소화기관내 불균형을 개선하고 유해균을 억제하며, 자연방어 효과를 강화시켜 체내 면역 반응을 향상시키는 기능을 한다. 그동안 장 면역 기능 향상을 위한 프로바이오틱스의 기능연구는 락토바실러스 (Lactobacillus), 락토코커스 (Lactococcus), 비피도박테리움 (Bifidobacterium) 등을 이용하여 종 (species)과 속 (genus)의 분류학적 형태에 따라 연구되어 왔다.The human intestine has an important function of absorbing nutrients through digestion of food and excreting unnecessary substances out of the body due to the symbiosis of various kinds of bacteria. In addition, changes in the environment caused by intestinal bacteria are directly related to intestinal immunity, which in turn affects the body's immunity. As research on these microorganisms beneficial to intestinal health has been continued for many years, research results on the intestinal function, immunity and metabolic activity of lactic acid bacteria have been actively published around the world. Probiotics, which refer to living microorganisms, improve the imbalance in the digestive tract, suppress harmful bacteria, and enhance the natural defense effect to enhance the body's immune response. Meanwhile, chapter probiotics function studies for improving the immune function is Lactobacillus bacteria (Lactobacillus), Lactococcus (Lactococcus), by using a Bifidobacterium (Bifidobacterium) according to the taxonomic types of species (species) and genus (genus) Has been studied.
유산균의 연구는 1900년대 매치니코프가 유산균에 의한 수명 연장 효과를 발표하면서부터 시작되었으며, 1946년 Streptococcus pyogenes과 Serratia marcescen을 골육종 (Osteosarcoma) 환자를 대상으로 종양 내 직접 주입하였을 때에 일부의 환자들에게서 종양 치료 반응이 나타난 것을 바탕으로 유산균에 의한 항암 치료에 대한 연구가 본격화되어 많은 전임상 임상 연구들이 지금까지도 활발히 진행되고 있다.Lactobacillus research began in the 1900s when Matchnikov announced the effect of prolonging lifespan by lactobacilli. In 1946, when Streptococcus pyogenes and Serratia marcescen were injected directly into the tumor in osteosarcoma patients, some patients were treated. Based on the tumor treatment response, studies on anticancer treatment by lactic acid bacteria have been in full swing, and many preclinical clinical studies are still actively progressing to this day.
2016년 El-Nezami 연구진은, L. rhamnosus GG, E. coli Nissle 1917 그리고 열처리한 VSL#3을 일정비율로 섞어서 마우스 동종 종양모델에 경구 투여하였을 때, 항암제인 Cisplatin 과 비교하여 신혈관생성과정 억제를 통한 항암 효과가 있음을 밝혔다. 또한, 항암제를 투여한 그룹은 항암제의 독성으로 인한 마우스의 몸무게 감소가 나타난 반면에 프로바이오틱스를 투여한 그룹에서는 나타나지 않음을 입증함으로써 프로바이오틱스가 항암 치료제로서의 기능을 가질 수 있음을 입증하였다. 다만, 상기 연구들에서 사용된 프로바이오틱스는 여러 종의 복합 미생물이 함유되어 있는 것으로, 미생물 하나 하나에 대한 기능적인 연구에 대해서는 미흡한 부분이 있었다.In 2016, El-Nezami researchers, when mixed with L. rhamnosus GG, E. coli Nissle 1917, and heat-treated
암은 일반 세포들과 다르게 세포 증식능력이 무한하며, 그 증식 속도 또한 빠르게 이루어지고, 암세포를 둘러싼 혈관, 림프관 및 섬유아세포 등을 이용하여 종양미세환경을 형성하여 다른 조직으로의 전이 및 일반세포의 기능 상실 등을 유발함으로써 사망에 이르게 하는 무서운 질병 중 하나이다.Unlike normal cells, cancer has infinite cell proliferation ability, and its proliferation rate is also fast, and by forming a tumor microenvironment using blood vessels, lymphatic vessels, and fibroblasts surrounding cancer cells, metastasis to other tissues and normal cells It is one of the terrifying diseases that can lead to death by causing loss of function and the like.
이러한 암을 치료하고자 전세계의 연구자들이 암세포의 세포 내 신호기전 및 전이, 약물의 부작용 등에 관한 다양한 주제에 대하여 수많은 연구들을 진행하고 있으며, 매년 새로운 신약들이 개발되어 많은 암환자들에게 치료효과를 기대하게 하는 임상시험들이 실시되고 있다.In order to treat such cancer, researchers around the world are conducting numerous studies on various topics such as intracellular signaling mechanisms and metastasis of cancer cells, and side effects of drugs, and new drugs are developed every year to expect therapeutic effects to many cancer patients. Clinical trials are being conducted.
수술로 인한 종양 조직 제거와 방사선 치료 외에, 항암치료에 사용되고 있는 현재의 항암제들은 각 암 종에 따라 다르게 적용이 되고 있다. 일반적으로 암환자들에게 처방되고 있는 화학항암제들은 암세포를 표적으로 하는 것이 아니다 보니, 암세포를 죽이는 효과를 가지고 있으나 정상세포들에게도 영향을 미쳐 환자들에게서 탈모, 설사, 발열, 면역력 저하 등의 부작용이 발생되고 있다. 이후, 암에 대한 유전적인 연구 등을 바탕으로 각 암종에서 발생되는 유전변이를 타겟으로 하는 표적 항암제가 개발되어 기존의 화학항암제에 의해 발생하는 부작용은 많이 개선되었으나, 환경에 따른 적응이 매우 빠른 암세포가 표적 항암제의 공격으로부터 벗어나고자 항암제 내성을 유발하게 되어, 표적항암제에 의한 지속적인 암 치료효과를 100% 기대할 수 없다는 문제점이 있다.In addition to surgical removal of tumor tissue and radiation therapy, current anticancer drugs used for chemotherapy are applied differently depending on each cancer type. Since chemical anticancer drugs generally prescribed to cancer patients do not target cancer cells, they have the effect of killing cancer cells, but they also affect normal cells, causing side effects such as hair loss, diarrhea, fever, and lowering of immunity in patients. It is occurring. Since then, based on genetic research on cancer, target anticancer drugs that target genetic mutations occurring in each carcinoma have been developed, and the side effects caused by existing chemotherapy drugs have been greatly improved, but cancer cells that adapt very quickly to the environment. In order to escape from the attack of the target anticancer drug, it induces anticancer drug resistance, and thus there is a problem that 100% of the continuous cancer treatment effect by the target anticancer agent cannot be expected.
최근에는 항암제와 종양미세환경에 대한 연구가 활발히 이루어져 항암 효과는 유지하면서, 환자의 면역력을 조절하는 여러 면역 관문 억제제에 대한 면역항암제가 개발되어 환자들에게 치료제로 사용되고 있다. 그 중, PD-1/PD-L1에 대한 치료는 피부암, 폐암 등의 환자들에게 높은 치료 반응을 나타내는 것으로 알려져 있다. 이러한 면역항암제는 종양미세환경 내에서 면역 세포들의 기능을 조절함으로써 암세포의 증식 억제 및 면역 세포의 활성을 높이는 기능을 가지고 있다. 그러나, 면역항암제 또한 모든 환자들에게서 동일한 항암 효과를 나타내지 못하고, 면역항암제에 대한 biomarker가 뚜렷이 밝혀져 있지 않으며, JAK-STAT 유전자변이에 의한 항암제 내성이 발생하거나, 항체 합성물의 특성으로 인한 자가면역질환을 초래할 수 있으며, 고가의 치료비용이 발생하는 등의 문제점들이 존재한다.Recently, studies on anticancer drugs and tumor microenvironments have been actively conducted, and immuno-anticancer agents have been developed against various immune checkpoint inhibitors that regulate the patient's immunity while maintaining anticancer effects, and are being used as therapeutics for patients. Among them, it is known that treatment for PD-1/PD-L1 shows a high therapeutic response to patients such as skin cancer and lung cancer. These immune anticancer agents have a function of inhibiting the proliferation of cancer cells and increasing the activity of immune cells by regulating the function of immune cells in the tumor microenvironment. However, immune anticancer drugs also do not show the same anticancer effect in all patients, and biomarkers for immune anticancer drugs are not clearly identified, and anticancer drug resistance occurs due to JAK-STAT gene mutation, or autoimmune diseases due to the characteristics of antibody compounds. It can be caused, and there are problems such as expensive treatment costs.
이러한 연구 결과들을 토대로, 본 발명자들은 실질적으로 암환자에게 치료 효과 개선을 나타낼 수 있는 미생물들에 대한 연구를 진행하여 왔으며. 비피도박테리움 비피덤 MG731 (Bifidobacterium bifidum MG731) 균주가 암 세포의 증식 억제 등에 우수한 효과를 나타내는 사실을 발견함으로써 본 발명을 완성하게 되었다.Based on these research results, the present inventors have been conducting research on microorganisms that can substantially improve the therapeutic effect in cancer patients. The present invention was completed by discovering the fact that the Bifidobacterium bifidum MG731 strain exhibits an excellent effect on inhibiting the proliferation of cancer cells.
본 발명은 항암 및 항염증 효과를 나타내는 프로바이오틱스 또는 그의 추출물을 신규의 항암제, 염증성 질환 치료제 또는 면역질환 치료제 등의 개발에 활용하기 위한 것이다.The present invention is intended to utilize probiotics exhibiting anti-cancer and anti-inflammatory effects or extracts thereof for the development of novel anti-cancer agents, inflammatory diseases therapeutics, immune diseases therapeutics, and the like.
따라서, 본 발명의 하나의 목적은 우수한 암의 예방 또는 치료 효과를 갖는 비피도박테리움 비피덤 MG731 (Bifidobacterium bifidum MG731) 균주를 제공하는 것이다.Accordingly, one object of the present invention is to provide a Bifidobacterium bifidum MG731 (Bifidobacterium bifidum MG731) strain having an excellent cancer prevention or treatment effect.
본 발명의 다른 목적은 비피도박테리움 비피덤 MG731 (Bifidobacterium bifidum MG731) 균주를 포함하는 암의 예방 또는 치료용 약학 조성물을 제공하는 것이다.Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer comprising a strain of Bifidobacterium bifidum MG731 (Bifidobacterium bifidum MG731).
또한, 본 발명은 비피도박테리움 비피덤 MG731 (Bifidobacterium bifidum MG731) 균주를 포함하는 암의 예방 또는 개선용 식품 조성물 또는 동물용 사료 조성물을 제공하는 것을 목적으로 한다.In addition, it is an object of the present invention to provide a food composition for preventing or improving cancer or a feed composition for animals, including a strain of Bifidobacterium bifidum MG731 (Bifidobacterium bifidum MG731).
상기 목적을 달성하기 위하여, 본 발명은 신규한 비피도박테리움 비피덤 MG731 (Bifidobacterium bifidum MG731) 균주를 제공한다. 상기 균주는 2018년 01월 04일 한국생명공학연구원 생물자원센터에 수탁번호 KCTC13452BP로 기탁되었다.In order to achieve the above object, the present invention provides a novel Bifidobacterium bifidum MG731 (Bifidobacterium bifidum MG731) strain. The strain was deposited with the accession number KCTC13452BP at the Korea Research Institute of Bioscience and Biotechnology Biological Resource Center on January 04, 2018.
또한, 본 발명은 비피도박테리움 비피덤 MG731 균주를 포함하는 암의 예방 또는 치료용 약학 조성물을 제공한다. 구체적으로, 상기 비피도박테리움 비피덤 MG731 균주는 균주 자체, 균주의 배양액, 또는 균주를 파쇄하여 얻은 세포질 분획물 (Cytoplasmic fraction)을 포함하는 것을 의미할 수 있다.In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of cancer comprising the strain Bifidobacterium bifidum MG731. Specifically, the Bifidobacterium bifidum MG731 strain may mean including the strain itself, a culture medium of the strain, or a cytoplasmic fraction obtained by crushing the strain.
본 발명은 유효량의 비피도박테리움 비피덤 MG731 (Bifidobacterium bifidum MG731) 균주를 암의 예방 또는 치료를 필요로 하는 대상체 (subject)에게 투여하는 것을 포함하는, 상기 대상체에서 암을 예방 또는 치료하는 방법을 제공한다. 본원에서 사용된 용어 "대상체"는 인간 및 비-인간 동물을 포함한다. 비-인간 동물은 모든 척추동물, 예를 들어 포유류 및 비-포유류, 예컨대 비-인간 영장류, 양, 개, 소, 말 등을 포함한다. 또한, 본 발명은 암의 예방 또는 치료를 위한 비피도박테리움 비피덤 MG731 (Bifidobacterium bifidum MG731) 균주의 용도를 제공한다.The present invention provides a method for preventing or treating cancer in a subject, comprising administering an effective amount of Bifidobacterium bifidum MG731 strain to a subject in need thereof. to provide. As used herein, the term “subject” includes humans and non-human animals. Non-human animals include all vertebrates, for example mammals and non-mammals, such as non-human primates, sheep, dogs, cattle, horses, and the like. In addition, the present invention provides a use of the Bifidobacterium bifidum MG731 strain for the prevention or treatment of cancer.
특히, 본 발명의 비피도박테리움 비피덤 MG731 균주는 항암, 항염증, 항산화 및 면역증진 효과를 모두 나타내는 것을 특징으로 한다.In particular, the Bifidobacterium bifidum MG731 strain of the present invention is characterized in that it exhibits all anticancer, anti-inflammatory, antioxidant and immune enhancing effects.
본 발명에 있어서, 암은 흑색종, 편평세포암종, 유방암, 두경부암, 갑상선암, 연부조직육종, 골육종, 고환암, 전립선암, 난소암, 방광암, 피부암, 뇌암, 혈관육종, 비만세포종, 백혈병, 림프종, 간암, 폐암, 췌장암, 위암, 신장암, 대장암, 조혈 종양, 신경 모세포종, 표피암종 또는 이의 전이암일 수 있으며, 이에 제한되지 않는다. 바람직하게, 본 발명에 있어서 암은 폐암, 대장암, 위암, 유방암 또는 간암일 수 있다.In the present invention, the cancer is melanoma, squamous cell carcinoma, breast cancer, head and neck cancer, thyroid cancer, soft tissue sarcoma, osteosarcoma, testicular cancer, prostate cancer, ovarian cancer, bladder cancer, skin cancer, brain cancer, hemangiosarcoma, mast cell tumor, leukemia, lymphoma , Liver cancer, lung cancer, pancreatic cancer, gastric cancer, kidney cancer, colon cancer, hematopoietic tumor, neuroblastoma, epidermal carcinoma, or metastatic cancer thereof, but is not limited thereto. Preferably, the cancer in the present invention may be lung cancer, colon cancer, stomach cancer, breast cancer or liver cancer.
본 발명에 있어서, 염증성 질환은 골관절염, 류마티스 관절염, 통풍, 강직성 척추염, 건염, 건막염, 류마티스 열, 루프스, 섬유근통, 건선 관절염, 천식, 아토피, 크론병 또는 궤양성 대장염일 수 있으며, 이에 제한되지 않는다.In the present invention, the inflammatory disease may be osteoarthritis, rheumatoid arthritis, gout, ankylosing spondylitis, tendonitis, tendonitis, rheumatoid fever, lupus, fibromyalgia, psoriatic arthritis, asthma, atopy, Crohn's disease, or ulcerative colitis, but is not limited thereto. Does not.
본 발명의 비피도박테리움 비피덤 MG731 균주는 암세포의 증식을 저해하고, 암세포의 이동성을 저하시킴으로써 항암 효과를 나타낼 수 있다. 또한, 본 발명의 비피도박테리움 비피덤 MG731 균주는 혈관생성인자인 VEGF (Vascular endothelial growth factor), Ang1 (Angiopoietin1), Ang2 (Angiopoietin2)의 발현을 억제함으로써 항암 효과를 나타낼 수 있다.The Bifidobacterium bifidum MG731 strain of the present invention may exhibit anticancer effects by inhibiting the proliferation of cancer cells and reducing the mobility of cancer cells. In addition, the Bifidobacterium bifidum MG731 strain of the present invention can exhibit an anticancer effect by inhibiting the expression of angiogenesis factors VEGF (Vascular endothelial growth factor), Ang1 (Angiopoietin1), and Ang2 (Angiopoietin2).
또한, 본 발명의 비피도박테리움 비피덤 MG731 균주가 TNF-α의 발현을 억제함으로써, 항염증 또는 항암 효과를 나타낼 수 있다. TNF-α는 인체 내 감염, 외상, 패혈증, 류마티스 관절염 등과 같은 만성 또는 급성 염증반응 시에 면역세포에서 분비되는 사이토카인으로, TNF-α의 농도가 증가하게 되면 세포내 지질 및 당 대사과정에 손상을 유발하게 된다. TNF-α는 세포 괴사를 유도하는 사이토카인으로 알려져 있으나, 지속적인 TNF-a의 자극이 세포에 전해지게 되면 오히려 세포내 대사에 미친 영향에 의해 종양 형성 유전자가 발생하게 되고, 세포의 비정상적인 증식이 발생하게 됨으로써 암의 유발을 촉진하게 된다는 연구 결과들이 보고된 바 있다.In addition, the Bifidobacterium bifidum MG731 strain of the present invention may exhibit anti-inflammatory or anti-cancer effects by inhibiting the expression of TNF-α. TNF-α is a cytokine secreted by immune cells during chronic or acute inflammatory reactions such as infection, trauma, sepsis, rheumatoid arthritis, etc., and when the concentration of TNF-α increases, intracellular lipids and sugar metabolism are damaged. Will cause. TNF-α is known as a cytokine that induces cell necrosis, but when continuous stimulation of TNF-a is transmitted to cells, tumorigenic genes are generated due to the effect on intracellular metabolism, and abnormal proliferation of cells occurs. There have been reports of research results that promote the induction of cancer by doing so.
본 발명은 비피도박테리움 비피덤 MG731 균주, 및 화학항암제 또는 면역항암제를 포함하는 것을 특징으로 하는, 암의 예방 또는 치료용 약학 조성물에 관한 것이다. 또다른 실시태양에서, 본 발명은 비피도박테리움 비피덤 MG731 균주를 포함하며, 화학항암제 또는 면역항암제와 병용 투여되는 것을 특징으로 하는, 암의 예방 또는 치료용 약학 조성물을 제공한다. 또한, 본 발명은 유효량의 비피도박테리움 비피덤 MG731 균주를 화학항암제 또는 면역항암제와 병용하여, 암의 예방 또는 치료를 필요로 하는 대상체에게 투여하는 것을 포함하는, 상기 대상체에서 암을 예방 또는 치료하는 방법을 제공한다. 상기 실시태양은 비피도박테리움 비피덤 MG731 균주를 화학항암제 또는 면역항암제와 함께 병용 투여하는 것을 특징으로 하는 것으로, 비피도박테리움 비피덤 MG731 균주와 화학항암제 또는 면역항암제를 하나의 조성물에 함께 포함하여 투여하는 것은 물론, 이들 각각이 별개로 포함된 조성물을 이를 필요로 하는 환자에게 투여하는 것을 포함한다. 이 때 비피도박테리움 비피덤 MG731 균주; 및 화학항암제 또는 면역항암제는 순차로 또는 동시에, 이를 필요로 하는 환자에게 투여될 수 있다.The present invention relates to a pharmaceutical composition for the prevention or treatment of cancer, characterized in that it comprises a Bifidobacterium bifidum MG731 strain, and a chemoanticancer agent or an immune anticancer agent. In another embodiment, the present invention provides a pharmaceutical composition for the prevention or treatment of cancer, comprising a strain of Bifidobacterium bifidum MG731, characterized in that it is administered in combination with a chemical anticancer agent or an immune anticancer agent. In addition, the present invention comprises administering an effective amount of Bifidobacterium bifidum MG731 strain to a subject in need of prevention or treatment of cancer by using in combination with a chemoanticancer agent or an immune anticancer agent, preventing or treating cancer in the subject. Provides a way to do it. The embodiment is characterized in that the Bifidobacterium bifidum MG731 strain is administered in combination with a chemoanticancer agent or an immune anticancer agent, comprising the Bifidobacterium bifidum MG731 strain and a chemotherapy or immunocancer agent together in one composition In addition to administering, it includes administering a composition each of which is separately included to a patient in need thereof. In this case, Bifidobacterium bifidum MG731 strain; And the chemo-anticancer agent or the immuno-cancer agent may be sequentially or simultaneously administered to a patient in need thereof.
또한, 본 발명은 유효량의 비피도박테리움 비피덤 MG731 균주를 화학항암제 또는 면역항암제와 병용하여, 암의 예방 또는 치료를 필요로 하는 대상체에게 투여하는 것을 포함하는, 상기 대상체에서 암을 예방 또는 치료하는 방법을 제공한다.In addition, the present invention comprises administering an effective amount of Bifidobacterium bifidum MG731 strain to a subject in need of prevention or treatment of cancer by using in combination with a chemoanticancer agent or an immune anticancer agent, preventing or treating cancer in the subject. Provides a way to do it.
상기 화학항암제는 옥살리플라틴 (Oxaliplatin), 페메트렉시드 (Pemetrexed), 시스플라틴 (Cisplatin), 젬시타빈 (Gemcitabine), 카보플라틴 (Carboplatin), 플루오로우라실 (5-FU), 싸이클로포스파마이드 (Cyclophosphamide), 파클리탁셀 (Paclitaxel), 빈크리스틴 (Vincristine), 에토포사이드 (Etoposide), 독소루비신 (Doxorubicin) 일 수 있으나, 이에 제한되지 않는다.The chemical anticancer agent is oxaliplatin, pemetrexed, cisplatin, gemcitabine, carboplatin, fluorouracil (5-FU), cyclophosphamide , Paclitaxel, Vincristine, Etoposide, Doxorubicin, but are not limited thereto.
또한, 상기 면역항암제는 면역관문 억제 기능을 갖는 anti-PD1, anti-PDL1, anti-CTLA, anti-Tim3, anti-LAG3일 수 있으나, 이에 제한되지 않는다.In addition, the anticancer agent may be anti-PD1, anti-PDL1, anti-CTLA, anti-Tim3, and anti-LAG3 having an immune checkpoint inhibitory function, but is not limited thereto.
또한 본 발명은 비피도박테리움 비피덤 MG731 (Bifidobacterium bifidum MG731) 균주를 포함하는 암의 예방 또는 개선을 위한 식품 조성물 또는 동물용 사료 조성물에 관한 것이다.In addition, the present invention relates to a food composition or animal feed composition for the prevention or improvement of cancer comprising a strain of Bifidobacterium bifidum MG731 (Bifidobacterium bifidum MG731).
상기 식품 조성물은 건강기능식품, 유제품, 발효제품 또는 식품첨가물일 수 있으며, 이에 제한되지 않는다.The food composition may be a health functional food, a dairy product, a fermented product, or a food additive, but is not limited thereto.
본 발명의 신규한 비피도박테리움 비피덤 MG731 균주는 다양한 암 세포주에 대하여 증식억제 효과를 갖는다. 또한, 본 발명의 비피도박테리움 비피덤 MG731 균주는 암세포의 이동성을 감소시키고, 신혈관의 생성을 억제하는 효과도 함께 가짐으로써, 기존에 알려진 다른 비피도박테리움 비피덤에 비하여, 월등하게 우수한 효과를 갖는다.The novel Bifidobacterium bifidum MG731 strain of the present invention has a proliferation inhibitory effect on various cancer cell lines. In addition, the Bifidobacterium bifidum MG731 strain of the present invention has the effect of reducing the mobility of cancer cells and inhibiting the generation of new blood vessels, so that it is superior to other known Bifidobacterium bifidum. Has an effect.
또한, 본 발명의 비피도박테리움 비피덤 MG731 균주는 항염증, 항산화 또는 면역증강 효과를 가짐으로써, 염증질환 또는 면역질환에도 사용될 수 있다.In addition, the Bifidobacterium bifidum MG731 strain of the present invention can be used for inflammatory diseases or immune diseases by having anti-inflammatory, antioxidant or immunity enhancing effects.
특히, 본 발명의 비피도박테리움 비피덤 MG731 균주는 단독으로 투여시 우수한 항암 효과를 나타낼 뿐만 아니라, 화학항암제 또는 면역항암제와 병용투여시 이들을 단독으로 투여하는 것과 비교하여 더욱 우수한 항암 효과를 갖는다.In particular, the Bifidobacterium bifidum MG731 strain of the present invention not only exhibits excellent anticancer effects when administered alone, but also has a more excellent anticancer effect compared to administering them alone when administered in combination with a chemical anticancer agent or an immune anticancer agent.
도 1은 정상인 154명과 125명의 폐암환자의 분변에서 장내 미생물 조성을 분석하기 위해 환자들의 병리학적 질병 상태에 따른 분류 및 치료 반응에 따른 분류를 나타낸 모식도이다.
도 2는 정상인과 폐암 환자들의 장내 미생물의 조성을 16s 서열 분석으로 분석하여 나타낸 것이다.
도 3은 폐암환자들 중 항암제 치료 반응에 따라 반응군과 비반응군 환자의 장내 미생물의 조성을 나타낸 것이다.
도 4는 장내 미생물 중, Bifidobacterium bifidum 유무에 따른 폐암환자들의 치료 반응을 나타낸 결과이다.
도 5는 인간 유래 암 세포주에서 MG731 처리에 따른 세포 증식 억제를 그래프로 나타낸 것이다.
도 6a 및 6b는 MG731에 의한 암세포주의 이동성 저하를 나타낸 결과이다.
도 7은 MG731이 신혈관생성 관련 인자인 VEGF의 발현을 저해한 것을 나타낸 것이다.
도 8a 및 8b는 VEGF 외의 신혈관생성 관련 인자인 Ang1, Ang2의 발현의 저해한 MG731의 효능을 확인한 결과이다.
도 9는 MG731이 LPS에 의한 염증반응 유발인자 중 TNF-α의 발현을 저해한 것을 나타낸 것이다.
도 10은 MG731의 농도별 처리에 따른 활성 산소 종의 감소를 나타낸 것이다.
도 11은 화학항암제 (Oxaliplatin, Pemetrexed)와 MG731의 병용 처리에 따른 암세포의 증식 억제효과를 나타낸 결과이다.
도 12는 도 11에서 염색된 세포들을 탈염색화하고 암세포의 증식 억제효과를 수치화하여 나타낸 것이다.
도 13은 인간의 혈액과 암세포주를 이용하여 MG731과 면역항암제 (anti-PD1, anti-PD-L1, anti-CTLA4)와의 병용 처리에 의한 암세포 사멸의 효능을 나타낸 것이다.
도 14은 마우스 동종 이식 모델을 이용하여 화학항암제 (Oxaliplatin)와 MG731의 병용 투여에 따른 종양 증식 억제효과를 나타낸 것이다.
도 15는 마우스 동종 이식 모델을 이용하여 면역항암제 (anti-PD1)와 MG731의 병용 투여에 따른 종양 증식 억제효과를 나타낸 것이다.
도 16는 화학항암제 (Oxaliplatin)와 MG731의 병용 투여에 따른 마우스 동종 이식 모델의 종양 조직 내 침투한 면역세포들의 분포를 분석한 것이다.
도 17은 면역항암제 (anti-PD1)와 MG731의 병용 투여에 따른 마우스 동종 이식 모델의 종양 조직 내 침투한 면역세포들의 분포를 분석한 것이다.1 is a schematic diagram showing classification according to pathological disease state and treatment response of patients in order to analyze the composition of intestinal microorganisms in feces of 154 and 125 normal patients with lung cancer.
Figure 2 shows the composition of the intestinal microbes of normal people and lung cancer patients analyzed by 16s sequence analysis.
3 shows the composition of intestinal microbes of the responding group and the non-responder group according to the response to treatment with an anticancer drug among lung cancer patients.
4 is a result showing the treatment response of lung cancer patients according to the presence or absence of Bifidobacterium bifidum among intestinal microorganisms.
5 is a graph showing the inhibition of cell proliferation according to MG731 treatment in human-derived cancer cell lines.
6A and 6B are results showing the decrease in the mobility of cancer cell lines by MG731.
7 shows that MG731 inhibited the expression of VEGF, a factor related to neovascularization.
8A and 8B are results confirming the efficacy of MG731 inhibiting the expression of Ang1 and Ang2, which are factors related to angiogenesis other than VEGF.
9 shows that MG731 inhibited the expression of TNF-α among the factors inducing the inflammatory response by LPS.
10 shows the reduction of active oxygen species according to the treatment of each concentration of MG731.
11 is a result showing the proliferation inhibitory effect of cancer cells according to the combination treatment of a chemical anticancer agent (Oxaliplatin, Pemetrexed) and MG731.
FIG. 12 shows the destaining of the cells stained in FIG. 11 and numerically shows the effect of inhibiting the proliferation of cancer cells.
13 shows the efficacy of cancer cell death by treatment in combination with MG731 and immuno-anticancer drugs (anti-PD1, anti-PD-L1, anti-CTLA4) using human blood and cancer cell lines.
14 shows the tumor proliferation inhibitory effect according to the combined administration of a chemoanticancer agent (Oxaliplatin) and MG731 using a mouse allograft model.
15 shows the tumor proliferation inhibitory effect according to the combined administration of an immuno-anticancer agent (anti-PD1) and MG731 using a mouse allograft model.
FIG. 16 is an analysis of the distribution of immune cells infiltrating into tumor tissues of a mouse allograft model according to the combined administration of a chemotherapy agent (Oxaliplatin) and MG731.
FIG. 17 is an analysis of the distribution of immune cells infiltrating into tumor tissues of a mouse allograft model according to the combined administration of an immunological anticancer agent (anti-PD1) and MG731.
본 발명자들은 우수한 항암 치료 또는 예방 효과를 갖는 프로바이오틱스를 발굴하기 위해 연구한 결과, 신규한 비피도박테리움 비피덤 MG731 균주가 우수한 항암 효과를 갖는 것을 확인하여 본 발명을 완성하였다.The present inventors have completed the present invention by confirming that the novel Bifidobacterium bifidum MG731 strain has an excellent anticancer effect as a result of researching to discover probiotics having an excellent anticancer treatment or prevention effect.
놀랍게도 MG731 균주는 폐암, 대장암, 위암, 유방암 및 간암 등의 다양한 암세포주에 대하여 우수한 암세포 증식억제 효과를 갖는다.Surprisingly, the MG731 strain has excellent cancer cell proliferation inhibitory effect against various cancer cell lines such as lung cancer, colon cancer, gastric cancer, breast cancer and liver cancer.
또한 암세포주에 MG731 균주를 처리한 경우, 암세포의 이동성이 현저히 저하된다. 암세포는 정상세포와 달리 세포에 손상이 발생하더라도 증식을 하면서 이동을 하는 특징이 있으며, 암세포의 이동성이 감소한다는 것은 암전이 가능성이 낮아진다는 것을 의미하므로 MG731 균주는 암전이 억제 효과를 갖는다.In addition, when the cancer cell line is treated with the MG731 strain, the mobility of cancer cells is significantly lowered. Cancer cells, unlike normal cells, are characterized by proliferating and migrating even if the cells are damaged, and decreasing the mobility of cancer cells means that the possibility of cancer metastasis is lowered, so the MG731 strain has an effect of inhibiting cancer metastasis.
또한, MG731 균주는 혈관생성 관련 주요 인자인 VEGF (Vascular endothelial growth factor), Ang1 (Angiopoietin1) 및 Ang2 (Angiopoietin2)의 발현을 모두 억제할 수 있다. 신혈관의 생성은 암세포의 특징 중 하나이며, 이를 억제하여 혈관을 통한 암세포로의 영양분 공급을 저해함으로써 암세포의 증식을 억제할 수 있다.In addition, the MG731 strain can inhibit all of the expressions of VEGF (Vascular endothelial growth factor), Ang1 (Angiopoietin1), and Ang2 (Angiopoietin2), which are major factors related to angiogenesis. The generation of new blood vessels is one of the characteristics of cancer cells, and by inhibiting them, it is possible to inhibit the proliferation of cancer cells by inhibiting the supply of nutrients to the cancer cells through blood vessels.
염증성 질환에서 뚜렷이 나타나는 양상 중의 하나는 활성 산소 종 (Reactive oxygen species; ROS)의 증가이다. 중간 정도의 활성 산소 종 농도는 세포 신호 전달 계통의 조절을 통해 효과를 발휘하지만, 실제로 높은 농도의 활성 산소 종에 장시간 노출되면 단백질, 지질 및 핵산에 비특이적인 손상을 초래하게 된다. 활성 산소 종은 단백질 인산화, 이온 채널 및 전사 인자의 산화 환원 조절과 같은 정상적인 생리적 과정에서 중요한 역할을 하며, 갑상선 호르몬 생성 및 세포 외 기질의 가교 결합을 포함한 생합성 과정에도 주요 기능을 가지고 있다. 또한, 대부분의 암세포에서도 이러한 활성 산소 종이 높은 활성을 가짐으로써 비정상적인 세포증식을 유발하는 것으로 널리 알려져 있다. 따라서, 본 발명의 MG731 균주는 활성 산소 종을 감소시켜 암세포의 증식을 억제하고, 여러 가지 질병의 발생을 예방하는 효과를 갖는다.One of the distinct patterns in inflammatory diseases is an increase in reactive oxygen species (ROS). A medium concentration of reactive oxygen species exerts an effect through the regulation of the cellular signaling system, but in fact, exposure to a high concentration of reactive oxygen species for a long time causes non-specific damage to proteins, lipids, and nucleic acids. Reactive oxygen species play an important role in normal physiological processes such as protein phosphorylation, redox regulation of ion channels and transcription factors, and have a major function in biosynthetic processes including thyroid hormone production and cross-linking of extracellular matrix. In addition, it is widely known that in most cancer cells, these reactive oxygen species have high activity, causing abnormal cell proliferation. Therefore, the MG731 strain of the present invention has an effect of reducing reactive oxygen species to inhibit the proliferation of cancer cells and prevent the occurrence of various diseases.
또한, 본 발명의 MG731 균주가 종양세포의 증식을 억제하는 기능을 가지는 Cytotoxic T cells (CD8+ effector T cells) 및 NK cells (Natural killer cells)을 종양 조직 내로 더 많이 침투하도록 유도하고, Cytotoxic T cells의 기능을 저해하는 T regulatory cells의 수를 감소시킴으로써, 면역세포의 기능을 조절하고 우수한 항암 효과를 나타낸다.In addition, the MG731 strain of the present invention induces more penetration of Cytotoxic T cells (CD8+ effector T cells) and NK cells (Natural killer cells) into tumor tissues, which have the function of inhibiting the proliferation of tumor cells, and By reducing the number of T regulatory cells that inhibit the function, it regulates the function of immune cells and exhibits excellent anticancer effects.
다양한 암세포주에 MG731 균주, 및 공지의 화학항암제 또는 면역항암제를 각각 처리하거나 병용 처리할 경우, MG731이 처리된 암세포주는 항암제가 처리된 암세포주 보다 우수한 세포 증식 억제 효과를 갖고, MG731 및 항암제와 병용 처리된 암세포주는 MG731 또는 항암제만 처리된 암세포주보다 뛰어난 세포 증식 억제 효과를 가지므로, MG731은 기존의 항암제와 병용 투여시 상승된 항암 효과를 갖는다.When various cancer cell lines are treated with or in combination with MG731 strain, and a known chemical anticancer or immune anticancer agent, cancer cell lines treated with MG731 have superior cell proliferation inhibitory effects than cancer cell lines treated with anticancer agents, and are used in combination with MG731 and anticancer agents. Since the treated cancer cell line has a superior cell proliferation inhibitory effect than that of MG731 or cancer cell lines treated with only anticancer agents, MG731 has an enhanced anticancer effect when administered in combination with an existing anticancer agent.
따라서, 본 발명의 비피도박테리움 비피덤 MG731 균주는 암세포 증식 억제, 암세포 이동성 저하 및 신혈관 생성 억제 효과를 동시에 나타내어 우수한 항암제로 활용될 수 있으며, 기존의 화학항암제 또는 면역항암제와 병용 투여될 수 있다.Therefore, the Bifidobacterium bifidum MG731 strain of the present invention can be used as an excellent anticancer agent by simultaneously exhibiting the effects of inhibiting cancer cell proliferation, lowering cancer cell mobility, and inhibiting new blood vessel generation, and can be administered in combination with an existing chemotherapy or immunotherapy. have.
본 발명의 비피도박테리움 비피덤 MG731 균주는 상기 화학항암제 또는 면역항암제와 하나의 제제로 동시에 투여되거나, 또는 별개의 제제로 동시에 또는 순차로 투여될 수 있다.The Bifidobacterium bifidum MG731 strain of the present invention may be administered simultaneously as a single agent with the chemical anticancer agent or immune anticancer agent, or may be administered simultaneously or sequentially as a separate agent.
또한, 염증유발인자인 LPS에 의해 유도된 TNF-α의 발현을 월등하게 감소시켜, MG731 균주는 염증성 질환 및 암을 동시에 예방 또는 치료할 수 있다.In addition, by remarkably reducing the expression of TNF-α induced by the inflammation-inducing factor LPS, the MG731 strain can prevent or treat inflammatory diseases and cancer at the same time.
본 발명은 비피도박테리움 비피덤 MG731 균주를 체내에 투여하여 암, 염증성 질환, 면역 질환 등을 예방 또는 치료하는 방법을 제공한다.The present invention provides a method for preventing or treating cancer, inflammatory diseases, immune diseases, etc. by administering the Bifidobacterium bifidum MG731 strain into the body.
본 발명의 비피도박테리움 비피덤 MG731 균주를 포함하는 조성물은 의약품, 건강기능식품, 유제품, 발효제품, 식품첨가물 또는 동물용 사료 등에 사용될 수 있다.The composition containing the Bifidobacterium bifidum MG731 strain of the present invention can be used in pharmaceuticals, health functional foods, dairy products, fermented products, food additives or animal feed.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 본 발명을 보다 구체적으로 설명하기 위한 것으로 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다.Hereinafter, the present invention will be described in more detail through examples. These examples are for explaining the present invention in more detail, and the scope of the present invention is not limited by these examples.
[실시예 1][Example 1]
임상환자 분변을 통한 장내 미생물 조성 분석Analysis of intestinal microbial composition through clinical patient feces
정상인 154명과 폐암환자 125명으로부터 분변을 제공받아 장내 미생물의 조성에 대한 연구를 위하여 16s sequence 분석을 시행하였다. 분석 결과에 대한 환자들의 병리학적 분류를 통해 장내 미생물 분석을 진행하고자 환자들의 차트에 따른 분류를 진행하였다. 125명의 환자의 종양조직에서, 가장 많이 발생하는 비소세포폐암 (Non-small cell lung cancer)에 속하는 폐암 2종인 편평상피암 (Squamous cell carcinoma)과 선암 (Adenocarcinoma) 환자들의 샘플만 분류한 다음, 환자들이 항암제의 치료를 받은 기록을 추적하여 항암제 반응에 따라 종양의 크기 변화를 측정한 수치를 기준으로 치료 반응 상태 (PD: Partial Response, SD: Stable Disease, PR: Progressive Disease)에 따른 환자의 샘플들을 분류하였다.Feces were provided from 154 normal subjects and 125 lung cancer patients, and 16s sequence analysis was performed to study the composition of intestinal microflora. In order to analyze the intestinal microflora through the pathological classification of the patients based on the analysis results, classification according to the patient chart was performed. In the tumor tissues of 125 patients, only samples of patients with squamous cell carcinoma and adenocarcinoma, two types of lung cancer, which are the most common non-small cell lung cancer, were classified. The patient's samples are classified according to the treatment response status (PD: Partial Response, SD: Stable Disease, PR: Progressive Disease) based on the value measured by measuring the size change of the tumor according to the response to the anticancer drug by tracking the record of receiving treatment with an anticancer drug. I did.
상기 기준에 따라, 비소세포폐암에 속하지 않는 폐암 환자들과 치료 반응 결과 추적이 불가능한 환자들에 속하는 49명의 샘플은 분석에서 제외되었다. 이에 대한 의약용어 및 상세한 설명은 도 1에 모식도로 표기하였다.According to the above criteria, samples of 49 patients belonging to lung cancer patients who do not belong to non-small cell lung cancer and patients whose treatment response results cannot be traced were excluded from the analysis. Medical terms and detailed descriptions thereof are shown in a schematic diagram in FIG. 1.
정상인과 폐암 환자들의 장내 미생물 환경에 있어, 정상인의 장내에 Bifidobacterium 속 균주들이 더 많이 분포되어 있음을 알 수 있으며 이에 대한 데이터는 도 2로 나타내었다.In the intestinal microbial environment of normal people and lung cancer patients, it can be seen that more strains of the genus Bifidobacterium are distributed in the intestine of normal people, and the data are shown in FIG. 2.
또한, 폐암 환자들의 항암 치료에 따라 반응군과 비반응군으로 분류하였을 때, 항암제 치료 반응이 있는 환자들의 장내 미생물에서 Bifidobacterium bifidum이 속해 있음을 확인하였으며, 이에 대한 분석은 도 3에 나타내었다.In addition, when the lung cancer patients were classified into responders and non-responders according to the anticancer treatment, it was confirmed that Bifidobacterium bifidum belongs to the intestinal microorganisms of patients with anticancer treatment reactions, and the analysis for this is shown in FIG.
이에 추가하여 항암 치료제에 따른 환자들의 치료 현황을 임상학적 분류로 표기하였을 때 Bifidobacterium bifidum이 장내 미생물 조성에 많을수록 환자들의 치료 반응이 더 좋음을 확인할 수 있었으며, 이는 도 4에 표기하였다. In addition, when the treatment status of patients according to the anticancer treatment was indicated by clinical classification, it could be confirmed that the more Bifidobacterium bifidum was in the composition of the intestinal microflora, the better the treatment response of the patients was, which is indicated in FIG. 4.
이러한 임상 환자들의 장내 미생물 분석을 통해, Bifidobacterium bifidum이 항암 치료 반응에 관계가 있음을 알 수 있다.Through the analysis of the intestinal microbiota of these clinical patients , it can be seen that Bifidobacterium bifidum is related to the response to chemotherapy.
[실시예 2][Example 2]
비피도박테리움 비피덤 MG731의 분리 및 배양Isolation and culture of Bifidobacterium Bifiderm MG731
MG731은 건강한 유아의 분변으로부터 Bifidobacterium spp.에 대한 선택배지를 이용하여 균주를 분리하였다.MG731 was isolated from the feces of healthy infants using a selective medium for Bifidobacterium spp.
수집한 분변 시료를 0.85％ NaCl에 10배 단계 희석하여 50 mg/L Lithium mupirocin을 첨가한 TOS-propionate agar (Merck KGaA, Darmstadt, Germany)에 도말하고 37℃의 온도에서 48시간 동안 혐기배양한 후 콜로니 (colony)의 형태가 서로 다른 균주를 선발하였다. 선발된 균주는 BL broth 에서 계대배양한 뒤 20% 글리세롤이 함유된 BL broth 에서 -80℃ 동결보관하였다.The collected fecal sample was diluted 10-fold in 0.85% NaCl, spread on TOS-propionate agar (Merck KGaA, Darmstadt, Germany) added with 50 mg/L Lithium mupirocin, and anaerobic cultured at 37°C for 48 hours. Strains having different colonies were selected. The selected strains were subcultured in BL broth and stored frozen at -80°C in BL broth containing 20% glycerol.
수득한 균주에 대한 rRNA의 염기서열을 분석하여 서열번호 1로 나타내고, 이를 동정한 결과 MG731은 비피도박테리움 비피덤 (Bifidobacterium bifidum)인 것을 확인하였다. 상기 MG731 균주는 2018년 1월 04일 한국생명공학연구원 생물자원센터에 수탁번호 KCTC13452BP 로 기탁되었다.The nucleotide sequence of the rRNA for the obtained strain was analyzed and represented by SEQ ID NO: 1. As a result of identifying it, it was confirmed that MG731 is Bifidobacterium bifidum . The MG731 strain was deposited with the accession number KCTC13452BP at the Korea Research Institute of Bioscience and Biotechnology Biological Resource Center on January 04, 2018.
본 실험에 사용하고자 하는 비피도박테리움 비피덤 MG731을 MRS broth (Difco, USA) 배지에 접종하고 37℃ 및 혐기성 조건 하에서 배양하면서 유산균의 증식이 OD=1 이 될 때 배양을 종료하였다. 배양이 종료된 배양물은 원심분리를 통해 균체를 회수하고, 회수된 균체는 PBS로 세척한 다음, PBS에 현탁하여 초음파 분쇄방법으로 균체를 분쇄하였다. 분쇄된 균체는 원심분리를 통해 상층액을 수득하여 0.42μm 필터를 통해 여과함으로써, 비피도박테리움 비피덤 MG731의 추출물을 제조하였다.Bifidobacterium bifidum MG731 to be used in this experiment was inoculated into MRS broth (Difco, USA) medium and cultured at 37° C. and under anaerobic conditions, and the cultivation was terminated when the proliferation of lactic acid bacteria reached OD=1. After the culture was completed, the cells were recovered through centrifugation, and the recovered cells were washed with PBS, then suspended in PBS, and the cells were pulverized by an ultrasonic grinding method. The pulverized cells were centrifuged to obtain a supernatant and filtered through a 0.42 μm filter to prepare an extract of Bifidobacterium bifidum MG731.
[실시예 3][Example 3]
비피도박테리움 비피덤 MG731Bifidobacterium Bifiderm MG731 균주의 종양 증식 억제 효과Inhibitory effect of strain on tumor proliferation
비피도박테리움 비피덤 MG731이 다양한 암세포주에서 항암 효능을 나타내는지 여부를 확인하기 위해서, 인간 유래 암 세포주를 이용하여 MTT assay를 시행하였다.In order to confirm whether Bifidobacterium bifidum MG731 exhibits anticancer efficacy in various cancer cell lines, MTT assay was performed using human-derived cancer cell lines.
MTT assay에 사용된 암 세포주로는 폐암 (A549, H1975, HCC827, H1299, SW900), 대장암 (HCT116, LoVo, SNU-C2A, SNU-C1, Colo205), 위암 (SNU216, AGS, MKN-28, MKN-1, SNU-601, SNU-1), 유방암 (Hs578T, BT20, MDA-MB-231, MCF7), 간암 (HepG2, Hep3B)으로 총 5종의 암 종을 이용하여 실험하였다.Cancer cell lines used in the MTT assay include lung cancer (A549, H1975, HCC827, H1299, SW900), colon cancer (HCT116, LoVo, SNU-C2A, SNU-C1, Colo205), gastric cancer (SNU216, AGS, MKN-28, MKN-1, SNU-601, SNU-1), breast cancer (Hs578T, BT20, MDA-MB-231, MCF7), and liver cancer (HepG2, Hep3B) were tested using a total of 5 carcinomas.
암 세포주를 96-well plate에 1~5×103 cells/well이 되도록 분주하고 24시간 후 유산균 시료를 1% (1% = 12.147μg, BCA 분석을 통해 추출물의 농도를 측정함) 로 첨가하여 72시간 배양한 다음, 각각의 well에 MTT (3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide thiazolyl blue) 시약을 처리하여 2시간 동안 반응시켰다.The cancer cell line was dispensed into a 96-well plate so as to be 1~5×10 3 cells/well, and after 24 hours, the lactic acid bacteria sample was added to 1% (1% = 12.147 μg, the concentration of the extract was measured through BCA analysis). After incubation for 72 hours, each well was treated with MTT (3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide thiazolyl blue) reagent and reacted for 2 hours.
이후, 살아있는 세포의 미토콘드리아에 반응하여 노란색인 MTT가 보라색으로 변하는 과정을 거쳤다. 이후 MTT가 들어간 배양액은 모두 제거하고 100 ㎕의 DMSO를 각 well에 첨가하여 보라색의 농도를 Microplate reader 장비를 사용하여 540nm 흡광도에서 측정하였고, 인간 유래 암세포주에 대한 실험결과를 도 5에 나타내었다.Then, in response to the mitochondria of living cells, the yellow MTT went through a process of turning purple. Thereafter, all the culture medium containing MTT was removed, and 100 µl of DMSO was added to each well, and the concentration of purple was measured at 540 nm absorbance using a Microplate reader equipment, and the experimental results for human-derived cancer cell lines are shown in FIG. 5.
[실시예 4][Example 4]
비피도박테리움 비피덤 MG731Bifidobacterium Bifiderm MG731 균주의 암세포 이동성 저하 효과The effect of reducing the mobility of cancer cells of the strain
암세포는 정상세포와 달리 세포에 손상이 오더라도 증식을 하면서 이동하는 특징이 있으므로, MG731 균주에 의해 암세포의 이동성이 저하되는지 여부를 확인하기 위하여, Wound healing assay를 시행하였다. A549 (5X105 cells) 및 HCT116 (6X105 cells) 균주를 6 well plate에 부착하여 세포가 90~95% 증식된 상태가 되면 tip을 이용하여 일정한 간격으로 세포에 손상을 가하였다. PBS 또는 MG731 균주를 24시간 동안 처리하여 세포의 이동성 여부를 현미경으로 관찰하였으며, 그 결과를 도 6에 나타내었다.Unlike normal cells, cancer cells are characterized by proliferating and migrating even if the cells are damaged, so a Wound healing assay was performed to determine whether the mobility of cancer cells was decreased by the MG731 strain. A549 (5X10 5 cells) and HCT116 (6X10 5 cells) strains were attached to a 6 well plate, and when the cells became 90-95% proliferated, the cells were damaged at regular intervals using a tip. PBS or MG731 strain was treated for 24 hours to observe the mobility of cells under a microscope, and the results are shown in FIG. 6.
도 6에 나타낸 바와 같이, 세포에 손상을 가한 A549 (도 6a)와 HCT116 (도 6b) 세포주의 0 시간대의 세포 상태를 기준으로 보았을 때, 24 시간 후에 A549는 65.31± 1.69 %, HCT116은 33.82 ± 5.86 % 암세포의 이동성이 활발히 나타난 반면, MG731을 처리한 A549와 HCT116에서는 각각 42.59 ± 4.01 % 및 22.63 ± 3.11 % 로 세포의 이동성이 저하되는 것을 확인할 수 있었다.As shown in FIG. 6, when the cell lines A549 (FIG. 6a) and HCT116 (FIG. 6b) damaged cells were observed based on the cell status at
결국, MG731 균주는 암 전이를 억제하여, 우수한 항암효과를 갖는 것을 알 수 있다.As a result, it can be seen that the MG731 strain inhibits cancer metastasis and has an excellent anticancer effect.
[실시예 5][Example 5]
비피도박테리움 비피덤 MG731Bifidobacterium Bifiderm MG731 균주의 신혈관생성 억제 효과Inhibitory effect of strain on neovascularization
암세포의 특징 중 하나인 신혈관생성 과정을 MG731 균주가 억제하는지 여부를 확인하기 위해서, 신혈관생성 관련 인자에 대한 발현 시험을 하기와 같이 시행하였다.In order to confirm whether the MG731 strain inhibits the neovascularization process, which is one of the characteristics of cancer cells, an expression test for angiogenesis-related factors was performed as follows.
HCT116 세포주에 MG731을 24시간 처리한 다음, RNA를 수득하여 cDNA를 합성하였으며, 이를 이용하여 혈관생성인자인 VEGF의 발현은 일반 PCR을, Ang1과 Ang2의 발현은 real-time PCR을 통해 확인하였으며, 그 결과를 도 7 및 8에 각각 나타내었다.After 24 hours treatment with MG731 in HCT116 cell line, RNA was obtained and cDNA was synthesized. Using this, the expression of VEGF, an angiogenesis factor, was confirmed through normal PCR, and the expression of Ang1 and Ang2 was confirmed through real-time PCR. The results are shown in Figs. 7 and 8, respectively.
도 7에 나타낸 바와 같이, MG731의 처리군에서 대조군과 대비하여 VEGF의 121 isoform 및 165 isoform의 발현이 현저히 감소한 것을 확인하였다.As shown in FIG. 7, it was confirmed that the expression of 121 isoform and 165 isoform of VEGF was significantly reduced in the MG731-treated group compared to the control group.
또한, 도 8에 나타낸 바와 같이, VEGF 이외에도 신혈관 생성에 관여하는 인자 Ang1 (도 8a) 및 Ang2 (도 8b)의 발현을 비교하였을 때 Ang1과 Ang2의 발현은 MG731의 처리군에서 대조군과 비교시 발현율이 70% 이상 현저히 저하되었음을 확인하였다.In addition, as shown in Figure 8, when comparing the expression of factors involved in the generation of new blood vessels in addition to VEGF Ang1 (Figure 8a) and Ang2 (Figure 8b), the expression of Ang1 and Ang2 was compared with the control group in the treatment group of MG731. It was confirmed that the expression rate was significantly decreased by more than 70%.
결국, MG731이 신혈관생성을 억제하여 혈관을 통한 암세포로의 영양분 공급을 저해함으로써 비정상적인 암세포의 증식 억제에 우수한 효과를 갖는 것을 알 수 있다.As a result, it can be seen that MG731 has an excellent effect on inhibiting the proliferation of abnormal cancer cells by inhibiting the generation of new angiogenesis and thus preventing the supply of nutrients to cancer cells through blood vessels.
[실시예 6][Example 6]
비피도박테리움 비피덤 MG731Bifidobacterium Bifiderm MG731 균주의 항염증 효과Anti-inflammatory effect of strain
MG731 균주의 항염증 효과를 확인하기 위해서, 마우스 대식세포인 RAW264.7 cells에 MG731 균주를 18시간 처리한 다음, 염증유발인자인 LPS 100 ng/ml를 6시간 처리하여 RNA를 수득하였다.In order to confirm the anti-inflammatory effect of the MG731 strain, the MG731 strain was treated for 18 hours in RAW264.7 cells, which are mouse macrophages, and then 100 ng/ml of LPS, which is an inflammatory factor, was treated for 6 hours to obtain RNA.
상기 RNA 1㎍으로 cDNA를 합성하였으며, 이를 통해 염증인자로 알려진 TNF-α의 발현여부를 real-time PCR을 통해 확인하였고, 그 결과를 도 9에 나타내었다.The cDNA was synthesized with 1 μg of the RNA, and the expression of TNF-α, which is known as an inflammatory factor, was confirmed through real-time PCR, and the results are shown in FIG. 9.
도 9에 나타낸 바와 같이, LPS에 의해 TNF-α의 발현이 증가한 정도를 상대지수 1로 하였을 때, MG731을 처리한 군은 TNF-α의 발현이 증가한 정도가 0.5 이하인 것을 확인하였다. TNF-α는 암을 유발하는 인자로도 알려져 있으므로, MG731에 의해 TNF-α의 발현이 현저히 감소된 것으로부터, MG731이 항염증 및 항암 기능을 동시에 갖는 것을 알 수 있다.As shown in FIG. 9, when the degree of increase in the expression of TNF-α by LPS was set to a relative index of 1, it was confirmed that the degree of increase in the expression of TNF-α was 0.5 or less in the group treated with MG731. Since TNF-α is also known as a cancer-causing factor, it can be seen that MG731 has both anti-inflammatory and anti-cancer functions from the remarkably reduced expression of TNF-α by MG731.
[실시예 7][Example 7]
비피도박테리움 비피덤 MG731Bifidobacterium Bifiderm MG731 균주의 항산화 활성 효과Antioxidant activity effect of strain
MG731의 항산화 활성을 확인하기 위하여, A549 암세포주에 MG731을 24시간 처리한 후, H2O2 0.5μM을 4시간 처리하고 DCFDA 형광 염료를 통해 FACs 장비로 세포내 활성 산소양을 측정하였으며, 그 결과를 도 10에 나타내었다.To confirm the antioxidant activity of MG731, after 24 hours treatment with MG731 on the A549 cancer cell line, 0.5 μM of H 2 O 2 was treated for 4 hours, and the amount of active oxygen in the cell was measured with FACs equipment through DCFDA fluorescent dye. The results are shown in FIG. 10.
도 10에 나타낸 바와 같이, PBS만 처리한 군은 4.66%의 활성 산소 양을 나타내었으며, H2O2만을 처리한 군은 64.8%로 활성산소가 증가한 것을 알 수 있었다. 반면, 0.1, 0.5, 1%의 MG731을 각각 처리한 군은 활성 산소의 양이 57.8%, 39.3% 및 32.5%로 감소되었다. 따라서, MG731이 활성산소의 양을 감소시키는 효과를 나타내고, MG731의 농도가 증가하면 활성 산소를 저해하는 효과도 증가하는 것을 확인하였다.As shown in FIG. 10, the group treated with only PBS showed an amount of active oxygen of 4.66%, and the group treated with only H 2 O 2 showed an increase in active oxygen of 64.8%. On the other hand, in the groups treated with 0.1, 0.5, and 1% of MG731, respectively, the amount of active oxygen decreased to 57.8%, 39.3% and 32.5%. Therefore, it was confirmed that MG731 showed an effect of reducing the amount of active oxygen, and when the concentration of MG731 increased, the effect of inhibiting active oxygen was also increased.
결국, 상기 실험결과를 통해 MG731은 항산화 효과를 갖는 것을 알 수 있다. 즉, 정상세포에서는 활성산소에 따른 세포 손상을 보호해주는 역할을 할 수 있고, 암세포에서는 비정상적인 활성 산소의 농도를 감소시켜 미토콘드리아의 비정상적인 기능을 제어하는 역할을 할 수 있다.As a result, it can be seen from the above experimental results that MG731 has an antioxidant effect. That is, in normal cells, it may play a role of protecting cell damage caused by free radicals, and in cancer cells, it may play a role of controlling abnormal functions of mitochondria by reducing the concentration of abnormal free radicals.
[실시예 8][Example 8]
비피도박테리움 비피덤 MG731Bifidobacterium Bifiderm MG731 균주와 항암제의 병용처리에 따른 암세포 증식 억제 효과 (Inhibitory effect of cancer cell proliferation by combination treatment of strain and anticancer agent ( in vitroin vitro 실험) Experiment)
A549, HCT116 에서 MG731과 항암제 병용 처리에 따른 암세포 증식 억제 실험을 실시하였다. 항암제로 Oxaliplatin 또는 Pemetrexed를 이용하여, 하기의 방법으로 실험을 수행하였다.In A549 and HCT116, cancer cell proliferation inhibition experiments were conducted by treatment with MG731 and anticancer drugs. Using Oxaliplatin or Pemetrexed as an anticancer agent, the experiment was performed in the following manner.
상기 두 가지의 암세포주를 6 well plate의 각 well에 1~2 X 103 이 되도록 희석 후 분주하여 24시간동안 부착시킨 후, 유산균과 항암제를 well 마다 처리하였으며, 2~3일 간격으로 배지를 교환해 주며 7일간 세포의 증식을 유도하였다. The two cancer cell lines were diluted to 1 ~ 2 X 10 3 in each well of a 6 well plate, dispensed and adhered for 24 hours, and then treated with lactic acid bacteria and anticancer drugs for each well, and the medium was added at intervals of 2 to 3 days. Exchange was performed and cell proliferation was induced for 7 days.
plate를 4% 포르말린으로 30분 동안 처리하여 세포를 고정시켜 세포 증식을 정지한 다음, 2번의 PBS 세척과정을 거쳐 crystal violet 용액으로 5분간 염색한 후 증류수로 세척하여 세포의 증식여부를 관찰하였다. 그 결과를 도 11에 나타내었다.The plate was treated with 4% formalin for 30 minutes to fix the cells to stop cell proliferation, followed by two PBS washing procedures, stained with a crystal violet solution for 5 minutes, and washed with distilled water to observe whether the cells proliferated. The results are shown in FIG. 11.
도 11에 나타낸 바와 같이, MG731 균주와 항암제를 병용 처리한 암 세포주에서 MG731 또는 항암제만 처리한 암 세포주보다 더 우수한 세포 증식 억제 효과가 나타나는 것을 알 수 있었다.As shown in FIG. 11, it was found that the cancer cell line treated with the MG731 strain and the anticancer agent together exhibited a superior cell proliferation inhibitory effect than the cancer cell line treated with only MG731 or an anticancer agent.
또한, 염색된 세포의 colony에 대한 수치는 Acetic acid로 Crystal violet을 녹여서 Microplate reader 기기를 이용하여 농도를 측정하였으며, 그 결과를 도 12에 나타내었다. 도 12에서도 도 11과 동일한 효과가 나타나는 것을 확인하였다.In addition, the concentration of the stained cell colony was determined by dissolving Crystal violet with Acetic acid and using a Microplate reader device, and the results are shown in FIG. 12. In FIG. 12, it was confirmed that the same effect as in FIG.
[실시예 9][Example 9]
비피도박테리움 비피덤 MG731Bifidobacterium Bifiderm MG731 균주와 면역 항암제의 병용투여에 따른 항종양 증진효과 (Anti-tumor enhancing effect of combined administration of strains and immune anticancer drugs ( in vitroin vitro 실험) Experiment)
인간의 혈액으로부터 Ficoll을 이용하여 PBMC (Peripheral blood mononuclear cells)를 수거한 후, RBC lysis buffer을 통해 적혈구를 제거하여 살아있는 세포수를 카운팅하여 유산균 (6 x 105/50ul/well)이 들어있는 바닥이 둥근 96웰 플레이트에 각 웰마다 PBMC 3x104 cells/50μl씩 첨가하여 24시간 배양하였다.After collecting PBMC (Peripheral blood mononuclear cells) from human blood using Ficoll, red blood cells are removed through RBC lysis buffer and the number of living cells is counted to the bottom containing lactic acid bacteria (6 x 10 5 /50ul/well). To this round 96-well plate, PBMC 3x10 4 cells/50 μl were added to each well and cultured for 24 hours.
결장암세포주 HCT116은 FBS가 포함되지 않은 RPMI배지에서 5 μM의 CFSE (Carboxyfluorescein succinimidyl ester)와 혼합하고, 37℃에서 5분 동안 반응시킨 다음, FBS를 포함한 RPMI1640 배지를 첨가하고, 10분 동안 ice에서 보관하였다. 상층액을 원심분리에 의해 제거한 후, 수득된 세포에 10% FBS가 함유된 RPMI1640 와 혼합한 후, 세포 수를 카운팅하여 상기 준비한 96웰 플레이트의 웰마다 3x104개/100μl씩 첨가하였다.Colon cancer cell line HCT116 was mixed with 5 μM of CFSE (Carboxyfluorescein succinimidyl ester) in RPMI medium without FBS, reacted at 37°C for 5 minutes, then RPMI1640 medium containing FBS was added, and stored on ice for 10 minutes. I did. After the supernatant was removed by centrifugation, the obtained cells were mixed with RPMI1640 containing 10% FBS, and then the number of cells was counted and 3×10 4 cells/100 μl were added to each well of the prepared 96-well plate.
이후, 암세포가 첨가된 웰마다 각 항체 PD1 (Pembrolizumab, A2005, Selleckem), PD-L1 (Atezolizumab, A2004, Selleckem), CTLA-4 (Ipilimumab, A2001, Selleckem) 20~30 ug/mL 농도 범위 안에서 처리하고 24시간 동안 배양한 다음, PBMC 및 암세포주의 혼합물에서 용균된 세포를 확인하기 위하여 7-아미노액티노마이신 D (7-AAD; BD Pharmingen, San Diego, CA, USA)로 세포를 염색하였다. FACSDiVa 소프트웨어 (BD Biosciences)를 이용하여 CFSE 및 7-AAD에 대한 염색을 측정함으로써 암세포주에 대한 PBMC의 세포용해능을 확인하여 그 결과를 도 13에 나타내었다.Then, each well to which cancer cells were added, each antibody PD1 (Pembrolizumab, A2005, Selleckem), PD-L1 (Atezolizumab, A2004, Selleckem), CTLA-4 (Ipilimumab, A2001, Selleckem) treated within the concentration range of 20 ~ 30 ug/mL And cultured for 24 hours, and then, cells were stained with 7-aminoactinomycin D (7-AAD; BD Pharmingen, San Diego, CA, USA) to identify cells lysed in a mixture of PBMC and cancer cell lines. By measuring staining for CFSE and 7-AAD using FACSDiVa software (BD Biosciences), the cytolytic ability of PBMCs against cancer cell lines was confirmed, and the results are shown in FIG. 13.
PBMC에 의한 암세포 사멸 (cytotoxicity)을 100% 기준으로 보았을 때, MG731 단독에 의한 암세포 사멸 (cytotoxicity)은 204.4% 증가, 임상에 사용되고 있는 면역항암제인 anti-PD1은 122.4%, anti-PD-L1은 133.6%, antiCTLA-4은 108%의 암세포 사멸을 유발하는 것으로 나타났다. 반면, 상기의 면역항암제와 MG731을 병용 처리한 경우, anti-PD1은 243.2%, anti-PD-L1은 221.6%, antiCTLA-4은 214.4%의 암세포 사멸을 유발하는 것으로 확인되었다.When looking at 100% of cancer cell death (cytotoxicity) caused by PBMC, cancer cell death (cytotoxicity) by MG731 alone increased by 204.4%, and anti-PD1, an immunological anticancer drug used in clinical practice, was 122.4%, and anti-PD-L1 was 133.6% and antiCTLA-4 were found to induce cancer cell death in 108%. On the other hand, it was confirmed that anti-PD1 induces 243.2% of cancer cell death, anti-PD-L1 in 221.6%, and antiCTLA-4 in 214.4% when treated with MG731.
결국, 단독 투여시의 암세포 사멸의 효과와 비교하여, 병용투여시 암세포 사멸의 효과가 월등하게 우수한 것을 알 수 있다.As a result, compared to the effect of killing cancer cells when administered alone, it can be seen that the effect of killing cancer cells when administered in combination is remarkably superior.
[실시예 10][Example 10]
비피도박테리움 비피덤 MG731Bifidobacterium Bifiderm MG731 균주와 항암제의 병용투여에 따른 종양 증식 억제 효과 (Inhibitory effect of tumor proliferation by co-administration of strain and anticancer drug ( in vivoin vivo 실험) Experiment)
종양 모델을 구축하기에 앞서 마우스에게 유산균 시료를 2주간 투여하여 장 정착 및 면역력을 높인 다음, 군당 C57BL/6 mouse 8마리의 오른쪽 엉덩이 부근에 2 X 105 MC38 cancer cells를 피하주사 함으로써 종양 유발 모델을 구축하였다. 종양 세포 주입과 동시에 3주간 (월요일~토요일) 유산균 시료를 상기 동물 모델에 경구 투여하였다. 투여한 유산균의 시료는 마리당 CFU 1 X 109이 되도록 200μl의 PBS에 희석하여 경구 투여하였다. 항암제로 옥살리플라틴 (3mg/kg, Sellekchem) 또는 anti-PD1 (2mg/kg, BioXCell)을 암 유발 후 매주 월요일과 목요일에 복강 주사하였다.Prior to constructing a tumor model, a lactic acid bacteria sample was administered to mice for 2 weeks to increase intestinal fixation and immunity, and then 2 X 10 5 MC38 cancer cells were injected subcutaneously in the vicinity of the right hip of 8 C57BL/6 mice per group. Was built. Simultaneously with tumor cell injection, a lactic acid bacteria sample was orally administered to the animal model for 3 weeks (Monday to Saturday). The sample of the administered lactic acid bacteria was diluted in 200 μl of PBS so that the CFU per head was 1 X 10 9 and administered orally. Anticancer agents oxaliplatin (3mg/kg, Sellekchem) or anti-PD1 (2mg/kg, BioXCell) were intraperitoneally injected every Monday and Thursday after cancer induction.
유산균만을 처리한 군, 옥살리플라틴 또는 anti-PD1만을 처리한 군, 및 둘 다를 처리한 군에 대해서 종양 억제 효과를 관찰한 후, 도 13 및 14에 나타내었다.After observing the tumor suppression effect in the group treated with only lactic acid bacteria, the group treated with only oxaliplatin or anti-PD1, and both treated, it is shown in FIGS. 13 and 14.
도 14 및 15에 나타낸 바와 같이, MG731을 단독으로 투여한 경우가 옥살리플라틴 또는 anti-PD1을 단독으로 투여한 경우보다 종양 억제 효과가 뛰어났으며, MG731을 옥살리플라틴 또는 anti-PD1과 병용 투여한 경우에는 종양 억제 효과가 더욱 우수한 것을 확인하였다.As shown in Figs. 14 and 15, when MG731 was administered alone, the tumor suppressing effect was better than when oxaliplatin or anti-PD1 was administered alone, and when MG731 was administered in combination with oxaliplatin or anti-PD1, It was confirmed that the tumor suppressing effect was more excellent.
[실시예 11][Example 11]
비피도박테리움 비피덤 MG731Bifidobacterium Bifiderm MG731 균주와 항암제의 병용투여에 따른 항종양 면역 반응 증진 효과 (Antitumor Immune Response Enhancement Effect of Co-administration of Strains and Anticancer Agents ( in vivoin vivo 실험) Experiment)
상기 실시예 10의 결과를 토대로, 종양 내에 침투한 면역세포의 분포를 확인하기 위해, FACs 실험을 시행하고자 다음과 같은 동물 실험을 진행하였다.Based on the results of Example 10, in order to confirm the distribution of immune cells infiltrating into the tumor, the following animal experiments were conducted to conduct FACs experiments.
실시예 10과 동일한 방법으로, 종양 유발 모델을 구축하고 유산균 시료 및 항암제를 투여한 후, 종양과 비장을 분리하여 마우스의 면역 세포 분포도를 확인하고자 조직을 파쇄하여 면역세포들을 분리하였다. 분리된 면역세포들은 각 기능에 따른 면역세포들의 마커에 해당하는 형광 항체를 이용하여 반응시킨 다음, FACs 장비를 이용하여 확인하였다. 상기 실험결과는 도 16과 도 17에 나타내었다.In the same manner as in Example 10, a tumor-inducing model was constructed, a lactic acid bacteria sample and an anticancer agent were administered, and then the tumor and the spleen were separated to determine the distribution of immune cells in the mouse, and the tissue was disrupted to separate immune cells. The isolated immune cells were reacted using fluorescent antibodies corresponding to markers of immune cells according to each function, and then confirmed using FACs equipment. The experimental results are shown in Figs. 16 and 17.
도 16 및 도 17에 나타낸 바와 같이, MG731을 투여한 그룹에서 항암 면역반응에 중요한 기능을 나타내는 CD4 T cells, CD8 T cells, CD8 effector T cells의 분포가 대조군인 IgG 혹은 PBS에 비하여 1.5~2 배 이상 증가하였음을 확인하였으며, T cells의 기능을 조절하는 regulatory T cells의 수는 현저히 감소되었음을 알 수 있다.As shown in FIGS. 16 and 17, the distribution of CD4 T cells, CD8 T cells, and CD8 effector T cells showing important functions in anticancer immune response in the group administered with MG731 was 1.5 to 2 times that of the control IgG or PBS. It was confirmed that the abnormality increased, and the number of regulatory T cells that regulate the function of T cells was remarkably decreased.
또한, MG731과 항암제가 병용 투여된 그룹에서는 항암제를 단독으로 투여한 그룹보다 CD4 T cells, CD8 T cells, CD8 effector T cells의 분포가 유의적으로 증가하며, T cells의 기능을 조절하는 regulatory T cells의 수는 유의적으로 감소한 결과를 통해 MG731이 항암제와 병용 투여가 되면 항암제 단독 처리보다 항종양 면역반응이 월등히 증가됨을 알 수 있다.In addition, the distribution of CD4 T cells, CD8 T cells, and CD8 effector T cells was significantly increased in the group administered with MG731 and an anticancer drug compared to the group administered with an anticancer drug alone, and regulatory T cells that regulate the function of T cells. The results showed that the antitumor immune response was significantly increased when MG731 was administered in combination with an anticancer agent compared to treatment with an anticancer agent alone.
상기 결과로부터 MG731에 의한 면역세포 기능의 조절이 종양 증식의 억제에도 영향이 미친다는 것을 알 수 있다.From the above results, it can be seen that the regulation of immune cell function by MG731 also affects the inhibition of tumor proliferation.
<110> GENOME AND COMPANY <120> BIFIDOBACTERIUM BIFIDUM MG731 AND COMPOSITION FOR PREVENTING OR TREATING CANCERS COMPRISING THE SAME <130> GNC19P-0002-KR-DIV <150> KR 10-2018-0054195 <151> 2018-05-11 <150> KR 10-2018-0133030 <151> 2018-11-01 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1447 <212> RNA <213> Artificial Sequence <220> <223> 16s rRNA Sequence of MG731 <400> 1 agacgctggc ggcgtgctta acacatgcaa gtcgaacggg atccatcggg ctttgcttgg 60 tggtgagagt ggcgaacggg tgagtaatgc gtgaccgacc tgccccatgc tccggaatag 120 ctcctggaaa cgggtggtaa tgccggatgt tccacatgat cgcatgtgat tgtgggaaag 180 attctatcgg cgtgggatgg ggtcgcgtcc tatcagcttg ttggtgaggt aacggctcac 240 caaggcttcg acgggtagcc ggcctgagag ggcgaccggc cacattggga ctgagatacg 300 gcccagactc ctacgggagg cagcagtggg gaatattgca caatgggcgc aagcctgatg 360 cagcgacgcc gcgtgaggga tggaggcctt cgggttgtaa acctcttttg tttgggagca 420 agccttcggg tgagtgtacc tttcgaataa gcgccggcta actacgtgcc agcagccgcg 480 gtaatacgta gggcgcaagc gttatccgga tttattgggc gtaaagggct cgtaggcggc 540 tcgtcgcgtc cggtgtgaaa gtccatcgct taacggtgga tctgcgccgg gtacgggcgg 600 gctggagtgc ggtaggggag actggaattc ccggtgtaac ggtggaatgt gtagatatcg 660 ggaagaacac cgatggcgaa ggcaggtctc tgggccgtca ctgacgctga ggagcgaaag 720 cgtggggagc gaacaggatt agataccctg gtagtccacg ccgtaaacgg tggacgctgg 780 atgtggggca cgttccacgt gttccgtgtc ggagctaacg cgttaagcgt cccgcctggg 840 gagtacggcc gcaaggctaa aactcaaaga aattgacggg ggcccgcaca agcggcggag 900 catgcggatt aattcgatgc aacgcgaaga accttacctg ggcttgacat gttcccgacg 960 acgccagaga tggcgtttcc cttcggggcg ggttcacagg tggtgcatgg tcgtcgtcag 1020 ctcgtgtcgt gagatgttgg gttaagtccc gcaacgagcg caaccctcgc cccgtgttgc 1080 cagcacgtta tggtgggaac tcacggggga ccgccggggt taactcggag gaaggtgggg 1140 atgacgtcag atcatcatgc cccttacgtc cagggcttca cgcatgctac aatggccggt 1200 acagcgggat gcgacatggc gacatggagc ggatccctga aaaccggtct cagttcggat 1260 cggagcctgc aacccggctc cgtgaaggcg gagtcgctag taatcgcgga tcagcaacgc 1320 cgcggtgaat gcgttcccgg gccttgtaca caccgcccgt caagtcatga aagtgggcag 1380 cacccgaagc cggtggccta accccttgtg ggatggagcc gtctaaggtg aggctcgtga 1440 ttgggac 1447 <110> GENOME AND COMPANY <120> BIFIDOBACTERIUM BIFIDUM MG731 AND COMPOSITION FOR PREVENTING OR TREATING CANCERS COMPRISING THE SAME <130> GNC19P-0002-KR-DIV <150> KR 10-2018-0054195 <151> 2018-05-11 <150> KR 10-2018-0133030 <151> 2018-11-01 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1447 <212> RNA <213> Artificial Sequence <220> <223> 16s rRNA Sequence of MG731 <400> 1 agacgctggc ggcgtgctta acacatgcaa gtcgaacggg atccatcggg ctttgcttgg 60 tggtgagagt ggcgaacggg tgagtaatgc gtgaccgacc tgccccatgc tccggaatag 120 ctcctggaaa cgggtggtaa tgccggatgt tccacatgat cgcatgtgat tgtgggaaag 180 attctatcgg cgtgggatgg ggtcgcgtcc tatcagcttg ttggtgaggt aacggctcac 240 caaggcttcg acgggtagcc ggcctgagag ggcgaccggc cacattggga ctgagatacg 300 gcccagactc ctacgggagg cagcagtggg gaatattgca caatgggcgc aagcctgatg 360 cagcgacgcc gcgtgaggga tggaggcctt cgggttgtaa acctcttttg tttgggagca 420 agccttcggg tgagtgtacc tttcgaataa gcgccggcta actacgtgcc agcagccgcg 480 gtaatacgta gggcgcaagc gttatccgga tttattgggc gtaaagggct cgtaggcggc 540 tcgtcgcgtc cggtgtgaaa gtccatcgct taacggtgga tctgcgccgg gtacgggcgg 600 gctggagtgc ggtaggggag actggaattc ccggtgtaac ggtggaatgt gtagatatcg 660 ggaagaacac cgatggcgaa ggcaggtctc tgggccgtca ctgacgctga ggagcgaaag 720 cgtggggagc gaacaggatt agataccctg gtagtccacg ccgtaaacgg tggacgctgg 780 atgtggggca cgttccacgt gttccgtgtc ggagctaacg cgttaagcgt cccgcctggg 840 gagtacggcc gcaaggctaa aactcaaaga aattgacggg ggcccgcaca agcggcggag 900 catgcggatt aattcgatgc aacgcgaaga accttacctg ggcttgacat gttcccgacg 960 acgccagaga tggcgtttcc cttcggggcg ggttcacagg tggtgcatgg tcgtcgtcag 1020 ctcgtgtcgt gagatgttgg gttaagtccc gcaacgagcg caaccctcgc cccgtgttgc 1080 cagcacgtta tggtgggaac tcacggggga ccgccggggt taactcggag gaaggtgggg 1140 atgacgtcag atcatcatgc cccttacgtc cagggcttca cgcatgctac aatggccggt 1200 acagcgggat gcgacatggc gacatggagc ggatccctga aaaccggtct cagttcggat 1260 cggagcctgc aacccggctc cgtgaaggcg gagtcgctag taatcgcgga tcagcaacgc 1320 cgcggtgaat gcgttcccgg gccttgtaca caccgcccgt caagtcatga aagtgggcag 1380 cacccgaagc cggtggccta accccttgtg ggatggagcc gtctaaggtg aggctcgtga 1440 ttgggac 1447
Claims (15)
Bifidobacterium bifidum MG731 strain (KCTC13452BP).
A pharmaceutical composition for the prevention or treatment of cancer, comprising Bifidobacterium bifidum MG731 strain (KCTC13452BP).
Bifidobacterium bifidum MG731 ( Bifidobacterium bifidum MG731) strain (KCTC13452BP), comprising the Bifidobacterium bifidum MG731 ( Bifidobacterium bifidum MG731) strain characterized in that it exhibits anti-cancer, anti-inflammatory, antioxidant and immunostimulating effects Pharmaceutical composition for the prevention or treatment of cancer.
According to claim 2, wherein the cancer is melanoma, squamous cell carcinoma, breast cancer, head and neck cancer, thyroid cancer, soft tissue sarcoma, osteosarcoma, testicular cancer, prostate cancer, ovarian cancer, bladder cancer, skin cancer, brain cancer, hemangiosarcoma, mast cell tumor, leukemia , Lymphoma, liver cancer, lung cancer, pancreatic cancer, gastric cancer, kidney cancer, colon cancer, hematopoietic tumor, neuroblastoma, epidermal carcinoma and metastatic cancer thereof, characterized in that at least one selected from the group consisting of, the pharmaceutical composition for preventing or treating cancer .
The pharmaceutical composition for preventing or treating cancer of claim 2, wherein the cancer is at least one selected from the group consisting of lung cancer, colorectal cancer, gastric cancer, breast cancer and liver cancer.
The pharmaceutical composition for preventing or treating cancer of claim 2, wherein the Bifidobacterium bifiderm MG731 strain exhibits an anticancer effect by lowering the mobility of cancer cells.
According to claim 2, wherein the Bifidobacterium Bifiderm MG731 strain exhibits an anticancer effect by inhibiting the expression of Vascular Endothelial Growth Factor (VEGF), Ang1 (Angiopoetin1) and Ang2 (Angiopoetin2), which are angiogenesis factors. Pharmaceutical composition for the prevention or treatment of cancer.
The pharmaceutical composition for preventing or treating cancer according to claim 3, wherein the Bifidobacterium bifidem MG731 strain inhibits the expression of TNF-α.
Bifidobacterium bifidum MG731 ( Bifidobacterium bifidum MG731) strain (KCTC13452BP), and characterized in that it further comprises a chemocancer or immunocancer drug, a pharmaceutical composition for the prevention or treatment of cancer.
10. The method of claim 9, wherein the chemotherapy agent is oxaliplatin (Oxaliplatin), Pemetrexed, Cisplatin (Cisplatin), Gemcitabine (Gemcitabine), Carboplatin (Carboplatin), Fluorouracil (5-FU), Cyclo Pharmaceutical composition for the prevention or treatment of cancer, characterized in that at least one selected from the group consisting of phosphamide (Cyclophosphamide), Paclitaxel, Vincristine, Etoposide and Doxorubicin (Doxorubicin).
The pharmaceutical composition for preventing or treating cancer of claim 9, wherein the immunocancer agent is selected from the group consisting of anti-PD1, anti-PDL1, anti-CTLA, anti-Tim3, and anti-LAG3.
10. The method for preventing or treating cancer according to claim 9, wherein the Bifidobacterium bifidum MG731 strain and the chemocancer or immunocancer agent are administered simultaneously in one formulation or simultaneously or sequentially in separate formulations. Pharmaceutical composition for.
Bifidobacterium bifidum MG731 ( Bifidobacterium bifidum MG731) comprising a strain (KCTC13452BP), characterized in that the pharmaceutical composition for the prevention or treatment of cancer, characterized in that administered in combination with a chemocancer or immunocancer.
A food composition for preventing or ameliorating cancer comprising Bifidobacterium bifidum MG731 strain (KCTC13452BP).
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102215599B1 (en) * | 2020-12-28 | 2021-02-15 | 주식회사 에이치이엠 | A novel strain of bifidobacterium animalis subsp. lactis hem20-01, and composition for treating depression comprising the strain or its culture fluid |
| KR102307603B1 (en) | 2020-11-13 | 2021-10-05 | 주식회사 비피도 | Composition with Bifidobacterium longum RAPO (KCTC13773BP) for preventing or treating cancer |
| KR102343938B1 (en) * | 2020-09-02 | 2021-12-28 | 주식회사 비피도 | Composition for improving inflammation containing recombinant Bifidobacterium bifidum BGN4 |
| KR20220152148A (en) | 2021-05-07 | 2022-11-15 | 주식회사 브이원바이오 | Limosillactobacillus reuteri VA102 and Anti-tumor composition using the same |
| KR102555685B1 (en) * | 2023-01-10 | 2023-07-14 | 주식회사 메디오젠 | Composition for preventing or treating gastric cancer containing a mixed strain of probiotics as an active ingredient |
-
2019
- 2019-06-12 KR KR1020190069415A patent/KR102040117B1/en active IP Right Grant
Non-Patent Citations (5)
| Title |
|---|
| Anaerobe, Vol.17, pp.226-231(2011.) * |
| Food Chem., Vol.135, pp.1914-1919(2012.) * |
| J. Dairy Sci., Vol.97, pp.7334-7343(2014.) * |
| Li J et al., Proc Natl Acad Sci USA. 2016 Mar 1;113(9):E1306-15 |
| Neoplasma, Vol.55, pp.387-393(2008.) * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102343938B1 (en) * | 2020-09-02 | 2021-12-28 | 주식회사 비피도 | Composition for improving inflammation containing recombinant Bifidobacterium bifidum BGN4 |
| KR102307603B1 (en) | 2020-11-13 | 2021-10-05 | 주식회사 비피도 | Composition with Bifidobacterium longum RAPO (KCTC13773BP) for preventing or treating cancer |
| KR102215599B1 (en) * | 2020-12-28 | 2021-02-15 | 주식회사 에이치이엠 | A novel strain of bifidobacterium animalis subsp. lactis hem20-01, and composition for treating depression comprising the strain or its culture fluid |
| KR20220152148A (en) | 2021-05-07 | 2022-11-15 | 주식회사 브이원바이오 | Limosillactobacillus reuteri VA102 and Anti-tumor composition using the same |
| KR102606078B1 (en) | 2021-05-07 | 2023-11-24 | 주식회사 브이원바이오 | Limosillactobacillus reuteri VA102 and Anti-tumor composition using the same |
| KR102555685B1 (en) * | 2023-01-10 | 2023-07-14 | 주식회사 메디오젠 | Composition for preventing or treating gastric cancer containing a mixed strain of probiotics as an active ingredient |
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