KR20200114996A - Faecalibacterium cancerinhibens and anti-cancer composition for colon cancer comprising the same - Google Patents

Abstract

The present invention provides a strain in the genus Faecalibacterium. The present invention also provides a pharmaceutical composition for preventing or treating cancer or a food composition for preventing or ameliorating cancer, including at least one selected from a group consisting of a fungus body of Faecalibacterium cancerinhibens strains having anticancer activity, a culture of the strains, a pulverized result of the strains, and an extract of the strains.

Description

Faecalibacterium cancerinhibens and anti-cancer composition for colon cancer comprising the same}

The present invention relates to a strain of the genus Pacalibacterium having anticancer activity, and an anticancer composition comprising the strain, a pharmaceutical composition for preventing or treating cancer, or a food composition for preventing or improving cancer.

Microbiome refers to microorganisms existing in a specific environment and the entire genetic information. In addition to the human body, microbiome information is being used in various fields such as animals, agriculture, the ocean, and the environment. In particular, with the development of human microbiome research based on advances in genetic information analysis and data analysis technology, The diagnostic industry and healthcare industry are expected to grow.

Human intestinal microbes not only decompose various substances that human enzymes cannot decompose and convert them into nutrients that can be absorbed by human cells, as well as inhibit the growth of externally-derived harmful bacteria to prevent pathogen infection. These intestinal bacteria themselves or various metabolites secreted by the bacteria stimulate numerous immune cells present in the intestinal cells to activate or regulate the human immune response. In addition, the human microbiome is regarded as the second genome of humans due to the number and diversity of symbiotic microbial genes that are more than 100 times that of humans, and its importance is recognized.

In particular, due to the development of human microbiome research, research results showing that intestinal microbes are directly or indirectly related to a number of human diseases are increasing rapidly, and research showing that intestinal microbes are also related to various cancers of the human body is also increasing. For example, in the case of colon cancer, an increase in bacteria of the genus Prevotella or Fusobacteria has been reported.

Colorectal cancer refers to malignant tumors occurring in the ascending colon, transverse colon, descending colon, sigmoid colon and rectal mucosa. Colorectal cancer includes adenocarcinoma, lymphoma, sarcoma, and squamous cell carcinoma, among which adenocarcinoma has the highest incidence. It is known that the incidence of colorectal cancer by region in Korea is ascending colon and s-phase colon, respectively, accounting for 25% and rectum accounting for 20%.

Looking at the statistics of colon cancer incidence in Korea, it was not a common disease before the 1970s, but in modern times, as dietary habits become westernized, the incidence of colon cancer is gradually increasing. According to statistics on the cause of death of the National Statistical Office (National Approval Statistics No. 10154) and cancer registration statistics from the Ministry of Health and Welfare (National Approval Statistics No. 11744), in the 10 years from 2000 to 2010, the incidence of colon cancer was 21.8 out of 100,000. At 41.5, the mortality rate nearly doubled from 8.8 out of 100,000 to 15.4.

As with other gastrointestinal cancers, surgical surgery is the most effective treatment for colorectal cancer at present, and the survival rate from the early stage to stage 3 reaches 70-90%, but the survival rate rapidly decreases to 15% in the last stage.

Conventional cancer treatment involves a combination of chemotherapy, surgery, hormone therapy, and/or radiation therapy to remove neoplastic cells in a patient. Cancer chemotherapy is preferably based on the use of drugs that kill replicating cells faster than the agent kills normal cells in the patient. Surgery is used to remove the tumor mass, but once the cancer has metastasized, it has little effect. Radiation is only effective in localized areas. All of these approaches present significant drawbacks and additional risks, such as increased susceptibility to infection.

Chemotherapy drugs often cause two complications that require treatment with antibiotics, which can eventually lead to dysbiosis: mucositis (a wound at the weakened mucosal gate associated with bacterial translocation) and neutropenia. do. Thus, there is a strong need for the development of improved cancer treatments that support constructive interactions between immunity and treatments such as chemotherapy and/or radiation, even if not synergistic effects.

An example of the present invention is to provide a strain of the genus Faecalibacterium having anticancer activity, for example, anticancer activity against colorectal cancer or colorectal cancer and liver cancer.

Another example of the present invention is a group consisting of an anticancer activity, for example, an anticancer activity against colorectal cancer, of a strain of the genus Faecalibacterium , a culture of the strain, a lysate of the strain, and an extract of the strain. It is to provide an anti-cancer composition comprising at least one selected from.

A further example of the present invention is a cell of the genus Faecalibacterium strain having anticancer activity, for example, anticancer activity against colon cancer or colon cancer and liver cancer, the strain culture, the lysate of the strain, and the strain It is to provide a pharmaceutical composition for the prevention or treatment of cancer, for example, colon cancer or colon cancer and liver cancer, including at least one selected from the group consisting of extracts of.

A further example of the present invention is a group consisting of an anticancer activity, for example, an anticancer activity against colorectal cancer, of a strain of the genus Faecalibacterium , a culture of the strain, a lysate of the strain, and an extract of the strain. To provide an anti-cancer food composition or anti-cancer functional food composition for colorectal cancer comprising at least one selected from.

A further example of the present invention is an anticancer activity, for example, colon cancer or tumor in a human individual by using the presence or presence of a strain of the genus Faecalibacterium having anticancer activity against colon cancer or colon cancer. It is to provide a method of obtaining useful information for detection and/or for predicting the efficacy of a drug in the therapeutic treatment of colon cancer or colon cancer, or tumor in a human subject.

In order to achieve the object of the present invention, the present inventors studied the genus Faecalibacterium , a genus constituting the major bacteria in the intestine, to isolate and identify a new strain, and the isolated strain of the genus Paecalibacterium was used as a cancer tumor in a mouse model. The present invention was completed by confirming that it has a production and proliferation inhibitory activity.

An example of the present invention relates to a strain of the genus Faecalibacterium having anticancer activity, specifically a strain of Faecalibacterium cancerinhibens , more specifically a strain of Faecalibacterium cancerinhibens , and more specifically a strain of Faecalibacterium cancerinhibens CLCC1. .

Another example of the present invention is an anticancer composition comprising at least one selected from the group consisting of the bacterial body of the strain, the strain culture, the lysate of the strain, and the extract of the strain, for example, colon cancer or colon cancer and It relates to an anticancer composition for liver cancer.

Another example of the present invention is for the prevention or treatment of colon cancer or colon cancer and liver cancer comprising at least one selected from the group consisting of the bacterial body of the strain, the strain culture, the lysate of the strain and the extract of the strain It relates to a pharmaceutical composition.

Another example of the present invention is an anticancer food composition for colon cancer or colon cancer and liver cancer comprising at least one selected from the group consisting of the bacterial body of the strain, the strain culture, the lysate of the strain, and the extract of the strain. It relates to a food composition for preventing or improving colon cancer or colon cancer and liver cancer or an anticancer functional food composition for colon cancer.

Another example of the present invention is colorectal cancer in a human individual by using the presence or presence of bacteria of the genus Faecalibacterium strain having anticancer activity, for example, anticancer activity against colon cancer or colon cancer and liver cancer. Or it relates to a method of obtaining useful information for detecting colorectal cancer and liver cancer or tumor and/or for predicting the efficacy of a drug in the therapeutic treatment of colorectal cancer or colorectal cancer and liver cancer or tumor in a human subject.

Hereinafter, the present invention will be described in more detail.

The present invention relates to a strain of the genus Faecalibacterium having anti-cancer activity, for example, anti-cancer activity against colorectal cancer or colorectal cancer and liver cancer, more specifically Faecalibacterium cancerinhibens CLCC1 strain. will be. The strain of the genus Pacalibacterium of the present invention may be isolated from the intestinal flora.

The strain of the genus Faecalibacterium having anticancer activity according to the present invention, more specifically the strain of Faecalibacterium cancerinhibens , has at least one or more of the following characteristics:

(1) Anticancer activity, a specific example, is an activity that delays the occurrence of a tumor or inhibits the growth rate of a tumor, and when the Faecalibacterium cancerinhibens CLCC1 strain is administered to a mouse subcutaneously transplanted with a cancer cell line, the control group does not administer the strain Activity with a tumor volume of 10 to 90% compared to,

(2) 16S rRNA having a nucleotide sequence of SEQ ID NO: 1 or a 16S rRNA comprising a nucleotide sequence having 97% or more nucleotide sequence identity thereto,

(3) culture temperature 20 to 40 ℃,

(4) Lipid production pattern, specifically, does not contain 17:0 iso 3OH fatty acids, including PL3, which is not present in Faecalibacterium prausnitzii , and

(5) Butyric acid production, in detail, has a content of 2 to 50 times higher than that of Faecalibacterium prausnitzii .

In more detail, the Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention is characterized by having anticancer activity, for example, anticancer activity against colon cancer. In addition to the anticancer activity against colon cancer, the strain may have additional anticancer activity against liver cancer.

In the present invention, "cancer" refers to a physiological condition in an animal, typically characterized by abnormal or uncontrolled cell growth. Cancer and cancer pathologies include, for example, metastasis, interference with normally functioning surrounding cells, release of cytokines or other secreted products at abnormal levels, suppression or augmentation of inflammatory or immunological responses, neoplasia, premalignancy. ), malignancy, surrounding or distant tissues or organs, such as lymph node invasion.

In the present invention, the cancer may be gastrointestinal cancer or non-gastrointestinal cancer.

The gastrointestinal cancer is a malignant tumor that occurs in the gastrointestinal tract, such as the esophagus, stomach, small intestine, or large intestine, and is found to be advanced enough to make complete resection of the cancer impossible in many cases because most of it has no subjective symptoms, and thus belongs to cancer with a poor prognosis. Accordingly, although various treatment methods have been developed with the recent development of medical technology, it belongs to cancer with a poor prognosis. In particular, gastric cancer in Korea ranks 1st among malignant tumors and ranks 1st in total cancer mortality, making it a very important disease not only for clinical but also for public health. In the case of early gastric cancer, the 5-year survival rate reaches 90 to 95%, and there is a need to develop a technology to slow the diagnosis and growth for early detection of cancer. The gastrointestinal cancer may be, for example, one or more cancers selected from the group consisting of esophageal cancer, gallbladder cancer, liver cancer, biliary tract cancer, pancreatic cancer, stomach cancer, small intestine cancer, colon cancer, colon cancer, anal cancer, and rectal cancer, but is not limited thereto. In an example, it may be colon cancer or colon cancer and liver cancer.

The non-gastrointestinal cancer includes, without limitation, malignant tumors occurring in organs other than the gastrointestinal tract or digestive system, for example, leukemia, lung cancer, prostate cancer, breast cancer, bladder cancer, kidney cancer, multiple myeloma, cervical cancer, thyroid cancer, ovarian cancer, urethra It may be cancer, skin cancer, osteosarcoma, glioblastoma, brain tumor, or lymphoma, but is not limited thereto.

In an embodiment of the present invention, the cancer may be colon cancer, and the colon cancer includes malignant tumors occurring at one or more sites selected from the group consisting of ascending colon, transverse colon, descending colon, sigmoid colon and rectal mucosa do. The colon cancer may be one or more types selected from the group consisting of adenocarcinoma, lymphoma, malignant carcinoma, leiomyosarcoma, Kaposi's sarcoma, and squamous cell carcinoma, but is not limited thereto.

The anticancer activity of the Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention may be an activity that inhibits the occurrence or progression of cancer. Specifically, it may be to delay the occurrence of a tumor or to inhibit the growth rate of the tumor, and may further have an activity of preventing and/or inhibiting metastasis of cancer due to the inhibition of the tumor growth rate. The tumor may be a tumor caused by colon cancer or a tumor caused by colon cancer and liver cancer.

In one embodiment of the present invention, the Faecalibacterium cancerinhibens CLCC1 strain was isolated from feces of a healthy adult male, and the effect of inhibiting tumor growth in a mouse subcutaneously implanted with a human colon cancer cell line was confirmed. As a result of comparing the tumor volume change of the three experimental groups administered with the Faecalibacterium cancerinhibens CLCC1 strain by concentration and the negative control group not administered the strain, the tumor volume in the experimental group was significantly reduced compared to the negative control group. Specifically, while the average tumor size for the current day (one day), start strain administration, but it seemed to 105mm ³ all four groups, after 36 days the average tumor volume from negative control appears to 4330mm ^3, CLCC1 strain is administered All of the experimental groups showed a volume of 3000 mm ³ or less, and it was confirmed to have a tumor volume of about 60 to 67% compared to the control group, and the effect of inhibiting the growth of colon cancer was confirmed by the Faecalibacterium cancerinhibens strain.

In one embodiment of the present invention, as a result of measuring the weight of the tumor and the weight of the mouse according to the administration of Faecalibacterium cancerinhibens CLCC1 in a mouse subcutaneously transplanted with a human colon cancer cell line, the weight of the mouse is irrespective of the administration of the strain. Although there was no significant difference, the weight of the tumor was significantly lower in all three experimental groups administered with the strain compared to the negative control group. Specifically, in the negative control group (G1), the weight of the tumor extracted after autopsy was about 3.21 g, but in the experimental group, the weight of the tumor tissue was markedly low, showing 1.94, 2.17 and 1.95 g at low, medium, and high concentrations, respectively.

Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention has a tumor growth inhibition rate of 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 5 to 90%, when the strain is administered, 5 to 70%, 5 to 50%, 10 to 90%, 10 to 70%, 10 to 50%, 15 to 90%, 15 to 70%, 15 to 50%, 20 to 90%, 20 to 70%, It may be 20 to 50%, 25 to 90%, 25 to 70%, 25 to 50%, 30 to 90%, 30 to 70%, or 30 to 50%. The tumor growth inhibition rate can be calculated by Equation 1 below.

In Equation 1, IR is a tumor growth inhibition rate (IR), T is an average tumor weight in each experimental group, and C is an average tumor weight in a negative control group.

In an embodiment of the present invention, Equation 1 above is used based on the results of measuring tumor weights in three experimental groups administered with the Faecalibacterium cancerinhibens CLCC1 strain at a low, medium or high concentration, and a negative control group without administration of the strain. Thus, the tumor growth inhibition rate was measured. In the experimental group administered with the above strain at a low concentration (2x10 ⁶ cells/head), the tumor growth inhibition rate was 39.6%, in the experimental group administered at a medium concentration (2x10 ⁷ cells/head), the tumor growth inhibition rate was 32.4%, and at a high concentration (2x10 ⁸ cells/head) head) showed a tumor growth inhibition rate of 39.3% in the experimental group, and it was confirmed that tumor growth was inhibited by more than 30% in all experimental groups.

In one embodiment of the present invention, the Faecalibacterium cancerinhibens CLCC1 strain was isolated from feces of a healthy adult male, and the effect of inducing liver cancer or inhibiting tumor growth in a transplanted mouse was confirmed. On average, compared to the control group, the volume of liver cancer tumors in the experimental group administered orally with CLCC1 was 19.5% to 75.3%, and the average value was 32.9% lower. Therefore, it can be seen that the orally administered CLCC1 strain has the activity of inhibiting the development and progression of tumors in liver cancer.

The Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention has a colorectal cancer tumor volume of 90% or less, 80% or less, 70 based on 100% of the colorectal cancer tumor volume of a negative control group not administered the strain when the strain is administered. % Or less, 67% or less, 10 to 90%, 10 to 80%, 10 to 70%, 10 to 67%, 20 to 90%, 20 to 80%, 20 to 70%, 20 to 67%, 30 to 90 %, 30 to 80%, 30 to 70%, 30 to 67%, 40 to 90%, 40 to 80%, 40 to 70%, 40 to 67%, 50 to 90%, 50 to 80%, 50 to 70 %, 50 to 67% or 60 to 67% level may have an activity that decreases.

When the strain of Faecalibacterium cancerinhibens CLCC1 provided by the present invention is administered, the tumor volume is 90% or less, 85% or less, 80% or less, 77 based on 100% of the volume of liver cancer tumors of the control group to which the strain was not administered. % Or less, 10 to 90%, 10 to 85%, 10 to 80%. Activity that decreases to a level of 10 to 77%, 15 to 90%, 15 to 85%, 15 to 80%, 15 to 77%, 18 to 90%, 18 to 85%, 18 to 80%, or 18 to 77% It may be to have.

The Faecalibacterium cancerinhibens strain provided by the present invention may be a 16S rRNA having a nucleotide sequence of SEQ ID NO: 1 or a 16S rRNA having a nucleotide sequence having 97% or more nucleotide sequence identity thereto. Specifically, it has at least 97%, 98%, 99%, 99.5%, 99.8%, 99.9% or 100% homology with the nucleotide sequence consisting of SEQ ID NO: 1 of the present specification.

The Faecalibacterium cancerinhibens strain provided by the present invention may be cultured at a temperature of 20 to 40° C., 25 to 40° C., 30 to 40° C., 35 to 40° C., or 37° C., characterized in that it is cultured under anaerobic conditions. I can.

Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention has a characteristic showing a lipid production pattern different from that of the existing Faecalibacterium genus strain Faecalibacterium prausnitzii .

Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention may be characterized in that it does not contain 17:0 iso 3OH fatty acid. In an embodiment of the present invention, as a result of analyzing the polar lipid content of the Faecalibacterium cancerinhibens CLCC1 strain and Faecalibacterium prausnitzii species, it was confirmed that CLCC1 additionally had PL3, which is not present in Faecalibacterium prausnitzii (FIG. 3B ).

Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention is 15:1 w8c fatty acid, 19:0 cyclo w10c/19w6 fatty acid, 20:1 w9c fatty acid, 14:0 3OH/16:1 iso I fatty acid, 15:0 3OH fatty acid, 16 It may include one or more fatty acids selected from the group consisting of :0 iso fatty acids and 16:0 iso 3OH fatty acids.

In one embodiment of the present invention, as a result of comparing the fatty acid composition of the Faecalibacterium cancerinhibens CLCC1 strain and the Faecalibacterium prausnitzii species by performing gas chromatography analysis, it was found that there is a difference in the composition of major fatty acids between the two strains. Specifically, 15:1 w8c fatty acid, 19:0 cyclo w10c/19w6 fatty acid, 20:1 w9c fatty acid, 14:0 3OH/16:1 iso I fatty acid, 15:0 3OH fatty acid, 16:0 iso fatty acid and 16: 0 iso 3OH fatty acid was detected only in CLCC1 strain, and 17:0 iso 3OH fatty acid was detected only in Faecalibacterium prausnitzii ATCC 27768 strain.

The Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention has a high production amount of butyric acid among short chain fatty acids (SCFA). Specifically, the production of butyric acid of the Faecalibacterium cancerinhibens CLCC1 strain of the present invention is 2 to 50 times, 2 to 40 times, 2 to 30 times, 2 to 20 times, 2 to 15 times, 2 to 10 times, 5 compared to Faecalibacterium prausnitzii species. To 50 times, 5 to 40 times, 5 to 30 times, 5 to 20 times, 5 to 15 times, or 5 to 10 times.

In one embodiment of the present invention, as a result of SCFA analysis of the culture supernatant of Faecalibacterium cancerinhibens CLCC1 strain and Faecalibacterium prausnitzii species of the present invention, Faecalibacterium cancerinhibens CLCC1 strain showed a butyric acid content 7.8 times higher than that of Faecalibacterium prausnitzii species (FIGS. .

Preferred strains of the genus Faecalibacterium according to the present invention have been given accession number KCTC 13783BP in addition to the Korea Research Institute of Bioscience and Biotechnology Biological Resources Center on January 3, 2019, and the scientific name is Faecalibacterium cancerinhibens . to be.

When the Faecalibacterium cancerinhibens strain provided by the present invention is plated on agar medium and cultured, colonies within 3 mm or 2 mm are formed, and have a long rod-shaped cell shape when observed with an optical microscope. In one embodiment of the present invention, Faecalibacterium cancerinhibens has a genome size of 2.9Mbp, the number of protein coding regions (CDS) is 2695, the GC ratio is 56.1%, the number of rRNA genes is 21, the total number of tRNA genes. Were found to be 69. Faecalibacterium cancerinhibens CLCC1 according to the present invention was 7.8 times higher than that of Faecalibacterium prausnitzii , butyric acid (butanoic acid) was detected (Fig. 3c to 3d).

Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention is characterized by having anti-cancer activity, for example, anti-cancer activity against colon cancer. The anticancer activity of the Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention may be an activity that inhibits the occurrence or progression of cancer, for example, colon cancer. Specifically, it may be to delay the occurrence of a tumor or to inhibit the growth rate of the tumor, and may further have an activity of preventing and/or inhibiting metastasis of colon cancer due to the inhibition of the tumor growth rate. Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention may additionally have anticancer activity against liver cancer in addition to anticancer activity against colon cancer.

The Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention has a colorectal cancer tumor volume of 90% or less, 80% or less, 70 based on 100% of the colorectal cancer tumor volume of a negative control group not administered the strain when the strain is administered. % Or less, 67% or less, 10 to 90%, 10 to 80%, 10 to 70%, 10 to 67%, 20 to 90%, 20 to 80%, 20 to 70%, 20 to 67%, 30 to 90 %, 30 to 80%, 30 to 70%, 30 to 67%, 40 to 90%, 40 to 80%, 40 to 70%, 40 to 67%, 50 to 90%, 50 to 80%, 50 to 70 %, 50 to 67% or 60 to 67% level may have an activity that decreases.

When the strain of Faecalibacterium cancerinhibens CLCC1 provided by the present invention is administered, the volume of liver cancer tumors is 90% or less, 85% or less, 80% or less, 77% or less, 10 to 90% compared to the control group to which the strain is not administered. , 10-85%, 10-80%. 10 to 77%, 15 to 90%, 15 to 85%, 15 to 80%, 15 to 77%, 18 to 90%, 18 to 85%, 18 to 80%, or 18 to 77% reducing activity Can be.

In one embodiment of the present invention, the Faecalibacterium cancerinhibens CLCC1 strain was isolated from feces of a healthy adult male, and the effect of inhibiting tumor growth in a mouse subcutaneously implanted with a human colon cancer cell line was confirmed. As a result of comparing the tumor volume change of the three experimental groups administered with the Faecalibacterium cancerinhibens CLCC1 strain by concentration and the negative control group not administered the strain, the tumor volume in the experimental group was significantly reduced compared to the negative control group. Specifically, while the average tumor size for the current day (one day), start strain administration, but it seemed to 105mm ³ all four groups, after 36 days the average tumor volume from negative control appears to 4330mm ^3, CLCC1 strain is administered All of the experimental groups showed a volume of 3000 mm ³ or less, and it was confirmed to have a tumor volume of about 60 to 67% compared to the control group, and the effect of inhibiting the growth of colon cancer was confirmed by the Faecalibacterium cancerinhibens strain.

In one embodiment of the present invention, as a result of measuring the weight of the tumor and the weight of the mouse according to the administration of Faecalibacterium cancerinhibens CLCC1 in a mouse subcutaneously transplanted with a human colon cancer cell line, the weight of the mouse is irrespective of the administration of the strain. Although there was no significant difference, the weight of the tumor was significantly lower in all three experimental groups administered with the strain compared to the negative control group. Specifically, in the negative control group (G1), the weight of the tumor extracted after autopsy was about 3.21 g, but in the experimental group, the weight of the tumor tissue was markedly low, showing 1.94, 2.17 and 1.95 g at low, medium, and high concentrations, respectively.

Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention has a tumor growth inhibition rate of 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 5 to 90%, when the strain is administered, 5 to 70%, 5 to 50%, 10 to 90%, 10 to 70%, 10 to 50%, 15 to 90%, 15 to 70%, 15 to 50%, 20 to 90%, 20 to 70%, It may be 20 to 50%, 25 to 90%, 25 to 70%, 25 to 50%, 30 to 90%, 30 to 70%, or 30 to 50%. The tumor growth inhibition rate can be calculated by Equation 1 below.

[Equation 1]

In Equation 1, IR is a tumor growth inhibition rate (IR), T is an average tumor weight in each experimental group, and C is an average tumor weight in a negative control group.

In an embodiment of the present invention, Equation 1 above is used based on the results of measuring tumor weights in three experimental groups administered with the Faecalibacterium cancerinhibens CLCC1 strain at a low, medium or high concentration, and a negative control group without administration of the strain. Thus, the tumor growth inhibition rate was measured. In the experimental group administered with the above strain at a low concentration (2x10 ⁶ cells/head), the tumor growth inhibition rate was 39.6%, in the experimental group administered at a medium concentration (2x10 ⁷ cells/head), the tumor growth inhibition rate was 32.4%, and at a high concentration (2x10 ⁸ cells/head) head) showed a tumor growth inhibition rate of 39.3% in the experimental group, and it was confirmed that tumor growth was inhibited by more than 30% in all experimental groups.

In one embodiment of the present invention, the Faecalibacterium cancerinhibens CLCC1 strain was isolated from feces of a healthy adult male, and the effect of inducing liver cancer or inhibiting tumor growth in a transplanted mouse was confirmed.

In an embodiment of the present invention, when liver cancer is caused through a subcutaneous tumor transplantation experiment of a liver cancer cell line, and the Faecalibacterium cancerinhibens CLCC1 strain is orally administered in the course of liver cancer for 55 days, in the experimental group to which the Faecalibacterium cancerinhibens CLCC1 strain is orally administered. Compared to the control group, it was confirmed that tumor occurrence and progression were statistically significantly inhibited (FIGS. 4A to 4C ). On average, compared to the control group, in the experimental group administered with CLCC1 orally, the tumor volume was 19.5% to 75.3%, and the average value was 32.9% lower. Therefore, it can be seen that the orally administered CLCC1 strain has an activity of inhibiting tumor development and progression.

In one embodiment of the present invention, when generating the liver in mice by transplantation experiments between the hepatoma cell line and, oral administration of Faecalibacterium spp CLCC1 in carcinogenesis process for 39 days, CLCC1 in the experimental group orally administered CLCC1 strain It was confirmed that cancer incidence and progression were statistically significantly suppressed compared to the control group not administered (FIGS. 5A to 5C ). Therefore, it can be confirmed that the CLCC1 strain has excellent tumor incidence and growth inhibitory activity.

An example of the present invention is useful information for detecting colon cancer or tumor in an individual and/or an animal comprising the step of measuring the proportion (%) of the Faecalibacterium cancer inhibis strain in the total intestinal flora of the individual. It relates to a method of obtaining useful information for predicting the efficacy of a drug in the therapeutic treatment of colon cancer or tumor in an individual.

An example of the present invention is a method for diagnosing colorectal cancer in an individual and a formulation for measuring the ratio of the strain, comprising the step of measuring the ratio (%) of the Faecalibacterium cancer inhibins strain in the intestinal flora of the individual It relates to a composition for diagnosing colon cancer comprising.

The agent for measuring the ratio of the strain can be used without limitation as long as it has specificity for Faecalibacterium cancer inhibins, for example, one or more agents selected from the group consisting of primers, probes, antibodies, and aptamers. I can.

The step of measuring the proportion of bacterial strains may be measuring the proportion of bacterial strains from an intestinal sample isolated from an individual, for example, an intestinal biopsy or a fecal sample. The method of measuring the ratio of the strain may be appropriately selected and used by a person skilled in the art according to the technical common knowledge in the art. For example, quantitative polymerase chain reaction (qPCR), massively parallel sequencing, fluorescent phosphorus -It may be performed using a molecular method selected from the group consisting of drilling hybridization (FISH), microarray and PCR-ELISA, but is not limited thereto.

Another example of the present invention is a method for detecting colon cancer or colon cancer and liver cancer, such as screening, diagnosing, differential diagnosis and/or monitoring of the onset and progression of colon cancer or colon cancer and liver cancer in an individual It is about. In addition, the present invention relates to a method for predicting the therapeutic treatment efficacy of a drug for colorectal cancer or colorectal cancer and liver cancer in an animal subject, comprising the step of confirming the presence of Faecalibacterium cancerinhinbens CLCC1 in an intestinal sample of the subject.

An example of the present invention is in the group consisting of colon cancer or colon cancer and liver cancer prophylactic or therapeutic activity of the strain of the genus Faecalibacterium , the strain culture, the lysate of the strain and the extract of the strain It relates to an anti-cancer composition comprising at least one selected.

The strain of the genus Faecalibacterium according to the present invention may be, for example, Faecalibacterium cancerinhibens , and exhibits an activity of inhibiting the occurrence and proliferation of colon cancer or colon cancer and liver cancer. It has tumor formation and/or tumor growth retardation activity. The strain of the genus Pacalibacterium according to the present invention may further have an activity of reducing or reducing metastasis of cancer cells in addition to the tumor formation and/or growth retarding activity of the cancer cells.

Specifically, at least about 10%, 20%, 30 compared to the corresponding tumor size, number of cancer cells, or tumor growth rate in the same subject prior to treatment, or compared to the corresponding activity in another subject not receiving treatment. %, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 100% can have the activity of reducing the tumor size, the number of cancer cells, or the growth rate of the tumor. Specifically, the Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention has a colorectal cancer tumor volume of 90% or less, 80% based on 100% of the colorectal cancer tumor volume of a negative control group without administration of the strain when the strain is administered. Or less, 70% or less, 67% or less, 10 to 90%, 10 to 80%, 10 to 70%, 10 to 67%, 20 to 90%, 20 to 80%, 20 to 70%, 20 to 67%, 30 to 90%, 30 to 80%, 30 to 70%, 30 to 67%, 40 to 90%, 40 to 80%, 40 to 70%, 40 to 67%, 50 to 90%, 50 to 80%, It may have an activity that decreases to a level of 50 to 70%, 50 to 67% or 60 to 67%.

In addition, the Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention has a tumor volume of 90% or less, 85% or less, 80% or less, based on 100% of the liver cancer tumor volume of the control group to which the strain was not administered when the strain was administered, 77% or less, 10 to 90%, 10 to 85%, 10 to 80%. Activity that decreases to a level of 10 to 77%, 15 to 90%, 15 to 85%, 15 to 80%, 15 to 77%, 18 to 90%, 18 to 85%, 18 to 80%, or 18 to 77% It may be to have.

In one embodiment of the present invention, the mouse to which the Faecalibacterium cansa inhibins strain is administered has a tumor volume decrease of about 30 to 40% compared to the mouse to which the strain is not administered. The effect of reducing the size of the tumor and inhibiting the growth rate of the tumor was confirmed.

In one embodiment of the present invention, mice administered with the Faecalibacterium cancerinhibis strain have an average tumor volume decrease of 32.9% compared to mice not administered with the strain, resulting in a reduction in tumor size and growth of the tumor. The rate-inhibiting effect was confirmed.

In one embodiment of the present invention, subcutaneous tumor cells were transplanted into mice administered with the Faecalibacterium cancerinhibens CLCC1 strain and a control mouse not administered the strain, and the incidence and growth rate of liver cancer was confirmed using MR photographs. As a result, Faecalibacterium cancerinhibens CLCC1 strain In the case of administration, the volume of the liver cancer tumor was 32.9% lower than that of the control group, indicating a delay in the occurrence of cancer and a delay in the growth rate of the tumor.

In one embodiment of the present invention, after transplanting the liver cancer cell line Hep55.1c to the left lobe of the liver of an 8-week-old mouse to induce liver cancer, comparing the tumor volume of the experimental group administered with Faecalibacterium cancerinhibens strain and the control group without administration As a result, when the tumor size on the day of transplantation was set to 1, tumor growth in the experimental group administered with the Faecalibacterium cancerinhibens strain was significantly suppressed compared to the control group.

The strain of the genus Faecalibacterium according to the present invention is Faecalibacterium cancerinhibens or Faecalibacterium cancerinhibens CLCC1. A closely related strain such as a strain of the genus Faecalibacterium having a 16S rRNA sequence that is at least 97%, 99%, 99.5% or 99.9% identical to the 16S rRNA gene sequence of the Faecalibacterium cancerinhibens strain can also be used. Preferably, the strain of the genus Pacalibacterium of the present invention has a 16S rRNA gene base sequence that is at least 97%, 99%, 99.5% or 99.9% identical to the base sequence of SEQ ID NO: 1.

The composition of the present invention is for use in the prevention, treatment or amelioration of cancer. The composition of the present invention can be administered to a patient diagnosed with cancer or confirmed to be at risk of developing cancer. The examples demonstrate that administration of the compositions of the present invention can reduce tumor growth in a number of tumor models. The treatment of colorectal cancer using the composition of the present invention can reduce tumor size or slow tumor growth. In certain embodiments, the compositions of the invention are for use in patients with solid tumors. In certain embodiments, the compositions of the present invention are for use in preventing metastasis of gastrointestinal cancer by delaying the onset or growth of tumors in gastrointestinal cancer treatment. In certain embodiments, the composition of the present invention is used to treat one or more gastrointestinal cancers selected from the group consisting of esophageal cancer, gallbladder cancer, liver cancer, biliary tract cancer, pancreatic cancer, gastric cancer, small intestine cancer, colon cancer, colon cancer, anal cancer, or rectal cancer. To use. In certain embodiments, the compositions of the invention are for use in treating colon cancer or colon cancer and liver cancer.

The compositions of the present invention may be particularly effective when used in combination with an additional therapeutic agent. In certain embodiments, the present invention provides a composition comprising a strain of Faecalibacterium cancer inhibins species and an anticancer agent. In a preferred embodiment, the anticancer agent is an immune checkpoint inhibitor, targeted antibody immunotherapy, CAR-T cell therapy, oncolytic virus or cell proliferation inhibitor.

The anticancer activity of the Faecalibacterium cancerinhibens CLCC1 strain provided by the present invention may be an activity that inhibits the occurrence or progression of cancer. Specifically, it may be to delay the occurrence of the tumor or inhibit the growth rate of the tumor, and may further have an activity to prevent metastasis of cancer due to the inhibition of the growth rate of the tumor.

The anticancer agent according to the present invention alone or in other therapies such as surgery, radiation therapy, gene therapy, immunotherapy, bone marrow transplantation, stem cell transplantation, hormone therapy, targeted therapy, cryotherapy, ultrasound therapy, photodynamic therapy, chemotherapy, etc. Can be done together. Additionally, those at greater risk of developing a proliferative disease may receive treatment to inhibit and/or delay the onset of the disease.

The anticancer effect of the composition of the present invention may be effective when combined with a more direct anticancer agent. Thus, in certain embodiments, the present invention provides a composition comprising a bacterial strain of Faecalibacterium cancerinhibens CLCC1 and an anticancer agent.

In the composition, the anticancer agent is as described above.

The active ingredient of the composition, at least one selected from the group consisting of the strains of the genus Pacalibacterium, the strain culture, the lysate of the strain, and the extract of the strain, is from 0.00001% to 100% by weight, from 0.001% by weight to 99.9% by weight, 0.1% by weight to 99% by weight, more preferably 1% by weight to 50% by weight.

The strain of the genus Pacalibacterium may be in a cell-free form that includes the strain or does not include the strain. The lysate refers to a lysate obtained by crushing the strains of the genus Pacalibacterium or a supernatant obtained by centrifuging the lysate. In the present specification, unless otherwise noted, the strain of the genus Pacalibacterium having anticancer activity is one selected from the group consisting of the cells of the strain, the culture of the strain, the lysate of the strain, and the extract of the strain. It is used to mean more.

The composition of the present invention may contain lyophilized bacteria. Freeze drying of bacteria can be performed by a person skilled in the art by a method known in the art. Alternatively, the composition of the present invention may comprise a culture of live active bacteria.

In some embodiments, the bacterial strain in the compositions of the present invention is not inactivated, eg, not heat-inactivated. In some embodiments, the bacterial strain in the composition of the present invention is not killed, eg, heat-killed. In some embodiments, the bacterial strain in the compositions of the present invention is not weakened, eg, heat-damaged. For example, in some embodiments, bacterial strains in the compositions of the invention are not killed and/or inactivated and/or weakened. For example, in some embodiments, the bacterial strain in the composition of the present invention is alive. For example, in some embodiments, the bacterial strain in the composition of the present invention is alive. For example, in some embodiments, bacterial strains in the compositions of the present invention are capable of partially or wholly colonizing the intestine. For example, in some embodiments, bacterial strains in the compositions of the present invention are viable and capable of partially or wholly colonizing the intestine.

The composition of the present invention comprises a therapeutically effective amount of a bacterial strain of the present invention. A therapeutically effective amount of the bacterial strain is sufficient to exert a beneficial effect on the patient. A therapeutically effective amount of the bacterial strain may be sufficient to cause intestinal transport and/or partial or full colonization of the patient. For example, a suitable daily dose of bacteria for adult humans may be from about 1 x 10 ³ to about 1 x 10 ¹¹ colony forming units (CFU); For example, from about 1 x 10 ⁷ to about 1 x 10 ¹⁰ CFU; In another example, it is about 1 x 10 ⁶ to about 1 x 10 ¹⁰ CFU.

An example of the present invention includes at least one selected from the group consisting of the bacteria of the genus Faecalibacterium strain, the strain culture, the lysate of the strain, and the extract of the strain having a cancer preventing or therapeutic activity. It relates to a pharmaceutical composition for the prevention or treatment of colon cancer or colon cancer and liver cancer.

The active ingredient of the composition, at least one selected from the group consisting of the strains of the genus Pacalibacterium, the strain culture, the lysate of the strain, and the extract of the strain, is from 0.00001% to 100% by weight, from 0.001% by weight to 99.9% by weight, 0.1% by weight to 99% by weight, more preferably 1% by weight to 50% by weight.

The pharmaceutical composition according to the present invention can be administered to mammals including humans by various routes. The mode of administration may be any method commonly used, for example, may be administered by oral, dermal, intravenous, intramuscular or subcutaneous routes, and preferably orally.

Typically, probiotics, such as the compositions of the present invention, are optionally combined with at least one suitable prebiotic compound. Prebiotic compounds are generally oligo- or polysaccharides or non-digestible carbohydrates such as sugar alcohols, which are not broken down or absorbed in the upper digestive tract. Known prebiotics include commercially available products such as inulin and transgalacto-oligosaccharides.

The composition of the present invention may contain a pharmaceutically acceptable excipient or carrier. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical field. Examples of suitable carriers include lactose, starch, glucose, methyl cellulose, magnesium stearate, mannitol, sorbitol and the like. Examples of suitable diluents include ethanol, glycerol and water. The choice of pharmaceutical carrier, excipient or diluent may be chosen with respect to the intended route of administration and standard pharmaceutical practice. The pharmaceutical composition may contain any suitable binders, lubricants, suspending agents, coatings, solubilizing agents as or in addition to carriers, excipients or diluents. Examples of suitable binders are starch, gelatin, natural sugars such as glucose, anhydrous lactose, free-flowing lactose, beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethyl cellulose and polyethylene glycol. Includes. Examples of suitable lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Preservatives, stabilizers, dyes and even flavoring agents can be provided in the pharmaceutical composition. Examples of preservatives include sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid. Antioxidants and suspending agents can also be used.

The dosage of the pharmaceutical composition of the present invention may vary depending on the patient's age, weight, sex, dosage form, health condition, and degree of disease, and may be divided into several times a day at regular intervals according to the judgment of the doctor or pharmacist. It can also be administered. For example, the daily dosage may be 0.1 to 500 mg/kg, preferably 0.5 to 300 mg/kg, based on the content of the active ingredient. The above dosage is an example of an average case, and the dosage may be high or low depending on individual differences.

The composition of the present invention may be encapsulated so as to be able to transport bacterial strains into the intestine. Encapsulation protects the composition from degradation until transported to the target site through rupture by chemical or physical stimuli, such as physical disruption that can be caused by, for example, changes in pressure, enzyme activity or pH. Any suitable encapsulation method can be used. Exemplary encapsulation techniques include entrapment in a porous matrix, adhesion or adsorption on the surface of a solid carrier, self-aggregation by agglomeration or crosslinking agents, and mechanical containment after a microporous membrane or microcapsule.

An example of the present invention includes at least one selected from the group consisting of the bacteria of the genus Faecalibacterium strain, the strain culture, the lysate of the strain, and the extract of the strain having a cancer preventing or therapeutic activity. It relates to a food composition or anti-cancer functional food composition for preventing or improving cancer.

In the present invention, food refers to a natural product or processed product containing one or more nutrients, and preferably refers to a state that can be eaten directly through a certain amount of processing process, and food, food in the usual sense It is intended to include all additives, health functional foods, beverages and beverage additives. In the present invention, a beverage refers to a generic term for drinking to quench thirst or to enjoy taste, and is intended to include a functional beverage. The beverage may be in liquid, syrup and/or gel form.

The content of the strain of the genus Pacalibacterium as an active ingredient contained in the food composition is not particularly limited appropriately depending on the form of the food and the desired use, for example, the strain of the genus Pacalybacterium, which is an active ingredient of the total food weight. At least one selected from the group consisting of cells, the strain culture, the lysate of the strain and the extract of the strain is 0.00001% to 100% by weight, 0.001% to 99.9% by weight, 0.1% to 99% by weight, more Preferably, it may be added in an amount of 1% to 50% by weight and 0.01 to 15% by weight. For example, the food composition may be added in an amount of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml.

The composition of the present invention can be formulated as a food product. For example, food can provide nutritional benefits in addition to the therapeutic benefits of the present invention, such as nutritional supplements. Similarly, a food composition can be formulated to be more attractive to consume the composition by enhancing the taste of the composition of the present invention or by more similar to a general food item than a pharmaceutical composition. In certain embodiments, the composition of the present invention is formulated as a milk-based product. The term "milk-based product" refers to a liquid or semi-solid milk or whey based product with varying fat content. Milk-based products are, for example, cow milk, goat milk, sheep milk, skim milk, whole milk, milk recombined from powdered milk and any non-processed milk or processed products such as yogurt, curds, curds, sour milk. (sour milk), prenatal milk, buttermilk, and other dairy products. Another important group includes milk beverages such as whey beverages, fermented milk, condensed milk, infant or baby milk; Flavored milk, ice cream; Includes milk-containing foods such as sweets.

One or more selected from the group consisting of the active ingredient of the composition, the cells of the Pacalybacterium cancer inhibins strain, the strain culture, the lysate of the strain, and the extract of the strain are 0.00001% to 100% by weight, 0.001% by weight % To 99.9% by weight, 0.1% to 99% by weight, more preferably 1% to 50% by weight.

In the anticancer composition according to the present invention, a pharmaceutical composition for preventing or treating colon cancer or colon cancer and liver cancer, or a food composition for preventing or improving colon cancer or colon cancer and liver cancer, the strain of the genus Pacalibacterium is as described above. .

The composition for the prevention or treatment of colon cancer or colon cancer and liver cancer comprising the strain of the genus Pacalibacterium according to the present invention has an activity of inhibiting the occurrence and growth of colon cancer or colon cancer and liver cancer, and in particular a pharmaceutical composition or It can be consumed by humans in the form of health functional foods, etc. and used for the prevention, treatment and/or improvement of colon cancer or colon cancer and liver cancer.

1A is a photograph showing a colony morphology formed after culturing Faecalibacterium cancerinhibens CLCC1 strain in Reinforced clostridial media (RCM) medium for 3 days.
1B is a photograph of the appearance of cells observed with an optical microscope in an exponenetial growth phase during liquid culture of Faecalibacterium cancerinhibens CLCC1 strain.
1C is a photograph of cells taken with a scanning electron microscope in an exponenetial growth phase during liquid culture of a strain of Faecalibacterium cancerinhibens CLCC1 strain.
Figure 2a is a picture showing the phylogenetic position obtained through 16S rRNA analysis of the Faecalibacterium cancerinhibens CLCC1 strain.
Figure 2b is a picture showing the phylogenetic position using 92 key genes present in the genome of the Faecalibacterium cancerinhibens CLCC1 strain.
2C is a diagram showing the relationship between the Faecalibacterium cancerinhibens CLCC1 strain and related species based on the mean nucleotide identity (ANI) value.
2D is a table comparing ANI values of Faecalibacterium cancerinhibens CLCC1 strain and related species.
3A is a graph showing the results of gas chromatography analysis comparing fatty acid composition of Faecalibacterium cancerinhibens CLCC1 strain and Faecalibacterium prausnitzii strain.
3B is a photograph showing the results of a two-dimensional chromatography analysis performed for comparison of polar lipids between the Faecalibacterium cancerinhibens CLCC1 strain and the Faecalibacterium prausnitzii strain.
Figure 3c is a GC-MS analysis graph of the short-chain fatty acid analysis of Faecalibacterium cancerinhibens CLCC1.
Figure 3d is This is a GC-MS analysis graph of short-chain fatty acid analysis of Faecalibacterium prausnitzii .
4A to 4C are graphs showing changes in tumor volume over time after induction of liver cancer in order to measure the inhibitory effect of CLCC1 strain on cancer induction in mice induced liver cancer by the method of Example 4. FIG. 4A is a point graph showing each individual with a sword, FIG. 4B is a graph comparing the average values of the experimental group and the control group, and FIG. 4C is a line graph showing changes in cancer volume for each individual.
5A to 5C are point graphs and line graphs of the results of a cancer incidence inhibitory effect test result of the CLCC1 strain in a liver transplantation model and each result. FIG. 5A is a result of directly comparing the size of a tumor, FIG. 5B is a result of expressing the change in tumor size in fold, and FIG. 5C indicates a delta value.
6A is a graph showing changes in tumor volume according to the dose of CLCC1 strain in mice subcutaneously transplanted with human colorectal cancer cell lines.
6B is a graph showing the results of measuring tumor weight according to the dose of the CLCC1 strain in mice subcutaneously transplanted with human colorectal cancer cell lines.
6C is a result of visual observation of a change in tumor size according to the dose of CLCC1 strain in a mouse subcutaneously transplanted with a human colorectal cancer cell line.
6D shows the results of visual observation by separating tumor tissues from the negative control group (G1) and each experimental group (G2 to G4) after the administration period of the CLCC1 strain in mice subcutaneously transplanted with a human colon cancer cell line.
6E is a result of performing H&E staining in order to confirm cell necrosis according to the administration of the CLCC1 strain in the tumor tissue of a mouse subcutaneously transplanted with a human colon cancer cell line.
6F is a result of performing TUNEL staining in order to confirm apoptosis according to administration of the CLCC1 strain in tumor tissues of mice subcutaneously transplanted with human colorectal cancer cell lines.

Hereinafter, the present invention will be described in more detail by examples. However, the scope of the present invention is not limited by these examples by the following examples.

Example 1. Isolation of strain

The bacteria of the genus Faecalibacterium are dominant bacteria in the intestines of healthy adults, accounting for about 10% of all intestinal bacteria, and are known to have direct or indirect effects on health.

Faecalibacterium cancerinhibens CLCC1 strain was isolated by diluting the feces of a healthy 20s adult male in Reinforced clostridial media (RCM) medium and then culturing it in an anaerobic culture condition at room temperature at 37°C.

Faecalibacterium cancerinhibens CLCC1 forms colonies of up to 2 to 3 mm when cultured on RCM agar medium for 3 days, and exhibits a long rod-shaped cell shape when observed with an optical microscope. 1A and 1B show the result of observation of the colony morphology and the morphology of cells using an optical microscope. Figure 1c shows a photograph of the shape of the cell using a scanning electron microscope.

Example 2. Phylogenetic analysis of strains

2-1. Faecalibacterium cancerinhibens Genetic sequencing of CLCC1 strain

The Faecalibacterium cancerinhibens CLCC1 strain isolated by the method of Example 1 was cultured in RCM liquid medium at room temperature under anaerobic culture conditions at 37° C. for 1 day, and then centrifuged to obtain a strain. The entire genome was isolated from the strain obtained using FastDNA SPIN Kit for Soil (MP Biomedicals).

After making a sequencing library using the PacBio sequencing library kit, whole genome sequencing was performed using the PacBio RS II platform, and genome sequencing information was analyzed using BIOiPLUG's Whole Genome analysis pipeline, our genome data analysis platform.

Table 1 below shows the genomic characteristics of the Faecalibacterium cancerinhibens CLCC1 strain. The total genome size of the Faecalibacterium cancerinhibens CLCC1 strain was about 2.9Mbp, the number of protein coding regions (CDS) was 2695, and the GC ratio (GC ratio, %) was 56.1%. There were 21 total rRNA genes and 69 total tRNA genes.

Item value Genome size (bp) 2,941,967 Protein coding site number (Number of CDSs) 2,695 GC ratio (GC ratio, %) 56.1 Number of rRNA genes 21 Number of tRNA genes 69

2-2. Phylogenetic analysis using 16S rRNA analysis

Phylogenetic taxonomic analysis was performed by analyzing the rRNA of the Faecalibacterium cancerinhibens CLCC1 strain sequence analyzed by the method of Example 2-1. The 16S rRNA sequence of the Faecalibacterium cancerinhibens CLCC1 strain is shown in SEQ ID NO: 1.

Specifically, similarity analysis of 16S rRNA was performed using the 16S-based identification for prokaryote pipeline of the EzBioCloud database, and phylogenetic analysis was performed using the MEGA program. Fig. 2a shows the phylogenetic position of the Faecalibacterium cancerinhibens CLCC1 strain identified by analyzing the 16S rRNA gene nucleotide sequence.

As a result of 16S rRNA similarity analysis, Faecalibacterium cancerinhibens CLCC1 strain showed 98% 16S rRNA similarity to Faecalibacterium prausnitzii ATCC 27768(T). The phylogenetic position of CLCC1 is "Firmicutes phylum, Clostridia river, Clostridiales order, Ruminococcaceae family.

2-3. Phylogenetic taxonomic analysis using key genes

Using the genome sequence data obtained by the method of Example 2-1, a phylogenetic taxonomic analysis was performed through gene sequence analysis.

Specifically, phylogenetic analysis was performed using the similarity of 92 genes (up-to-date bacterial core gene (UBCG)) that can be used as taxonomic markers of bacteria. The 92 genes refer to genes provided by the UBCG (up-to-date bacterial core gene) pipeline (Na, SI, Kim, YO, Yoon, SH, Ha, SM, Baek, I. & Chun, J. (2018)).

Fig. 2b shows a picture showing the phylogenetic position of the relative species obtained by analyzing 92 key genes.

2-4. Relationship analysis using Average Nucleotide Identity (ANI)

Using the genome analysis data obtained by the method of Example 2-1, the relationship was analyzed by comparing ANI values between the Faecalibacterium cancerinhibens CLCC1 strain and the related species.

Specifically, the genome similarity value was analyzed by comparing the entire genome sequence between the Faecalibacterium cancerinhibens CLCC1 strain and other related species using the Genome-based identification for prokaryote (TrueBAC ID) pipeline, an analysis platform provided by ChunLab.

Figure 2c shows the relationship between the Faecalibacterium cancerinhibens CLCC1 strain and related species based on the ANI value. Figure 2d shows a comparison table of the ANI values of the Faecalibacterium cancerinhibens CLCC1 strain and related species. Each row and column is 1 for Faecalibacterium cancerinhibens CLCC1, 2 for Faecalibacterium prausnitzii ATCC 27768(T), 3 for Fournierella massiliensis AT2(T), 4 for Intestinimonas butyriciproducens DSM 26588(T), 5 for Intestinimonas massiliensis GD2(T), 6 Pseudoflavonifractor capillosus ATCC 29799 (T), 7 is Ruthenibacterium lactatiformans 585-1 (T), 8 is Subdoligranul um variabile DSM 15176 (T), 9 is Gemmiger formicilis ATCC 27749 (T). (T) after each species name means type strain.

As a result of genome decoding of Faecalibacterium cancerinhibens CLCC1 strain and comparison with related species, CLCC1 strain showed an average nucleotide identity (ANI) value of 85.99% at the genome level with Faecalibacterium prausnitzil , the closest relative at the whole genome level. It can be seen that this is a novel species that has not been isolated and reported in. The strain of the genus Faecalibacterium identified by the above method was named Faecalibacterium cancerinhibens CLCC1.

Example 3. Analysis of fatty acid content of strain

In order to analyze the molecular biological properties of the Faecalibacterium cancerinhibens CLCC1 strain, the fatty acid composition of Faecalibacterium cancerinhibens CLCC1 strain and Faecalibacterium prausnitzii were compared.

Specifically, after culturing Faecalibacterium cancerinhibens CLCC1 strain and Faecalibacterium prausnitzii strain in RCM medium at 37° C. and anaerobic conditions, respectively, using about 40 mg of r cells, Miller's method (Miller, LT (1982) J. Clin. Microbiol. 18 , 861-867) to obtain a fatty acid.

Agilent technologies 6890 Gas chromatography was used for the analysis of the extracted fatty acids, and the separation column was A30m X 0.320mm X 0.25μm Crosslinked Methyl siloxane column (HP-1). 3A shows a graph of the results of gas chromatography analysis comparing the fatty acid composition of the Faecalibacterium cancerinhibens CLCC1 strain and the Faecalibacterium prausnitzii strain (ATCC 27768). In the graph of FIG. 3A, the names of fatty acids and their values (%, w/v) that each peak means It is shown in Table 2 below.

Type of fatty acid Content in CLCC1 (%) Content in ATCC 27768 (%) 16:00 32.42 42.28 18:1 w7 28.15 14.88 17:0 2OH 12.51 8.09 16:1 w7c/16:1 w6c 5.22 6.04 16:1 w9c 3.89 1.57 14:00 2.44 14.93 17:0 anteiso 2.42 0.99 16:1 w5c 2.17 0.83 18:1 w9c 1.88 0.86 18:00 1.59 2.17 16:1 2OH 1.41 0.51 20:1 w7c 0.84 0.43 18:1 w5c 0.81 0.32 15:0 2OH 0.75 0.5 18:0 10-methyl, TBSA 0.45 0.21 15:1 w8c 0.44 - 15:1 iso H/13:0 3OH 0.35 1.44 16:0 3OH 0.32 0.11 15:0 anteiso 0.21 0.2 12:00 0.2 1.59 15:0 iso 3OH 0.17 0.49 17:00 0.17 0.2 19:0 cyclo w10c/19w6 0.16 - 20:00 0.14 0.1 13:1 at 12-13 0.13 0.35 10:00 0.13 0.2 20:1 w9c 0.13 - 17:0 iso 0.12 0.17 14:0 3OH/16:1 iso I 0.12 - 15:0 3OH 0.11 - 16:0 iso 0.1 - 16:0 iso 3OH 0.05 - 17:0 iso 3OH - 0.54

As shown in Table 2 above, 15:1 w8c fatty acid, 19:0 cyclo w10c/19w6 fatty acid, 20:1 w9c fatty acid, 14:0 3OH/16:1 iso I fatty acid, 15:0 3OH fatty acid, 16:0 The iso fatty acid and 16:0 iso 3OH fatty acid were detected only in the CLCC1 strain, and the 17:0 iso 3OH fatty acid was detected only in the Faecalibacterium prausnitzii ATCC 27768 strain.

The molecular characteristics compared result, Faecalibacterium cancerinhibens CLCC1 strain was able to confirm that different species than Faecalibacterium prausnitzii species known to be included in Faecalibacterium cancerinhibens. When comparing the composition of the fatty acids of the two strains, it can be seen that there is a difference in the composition of the main fatty acids between the two strains, and the results are shown in FIGS. 3A and 2.

Example 4. Polar lipid analysis of strain

Thin layer chromatography (TLC) analysis was performed to compare the composition of polar lipids.

Specifically, polar lipids were extracted from a 50 mg strain lyophilized sample using Minikin's method (Minikin, D.E., et al. (1984). J Microbial Meth 2, 233-241.). And the first development was performed in a solvent (chloroform:methanol:water = 65:25:3.8 (v/v)) in a ratio of chloroform, methanol and water 65:25:3.8 (v/v). Thereafter, the second in a solvent (chloroform:methanol:acetic acid:water = 40:7.5:6:1.8 (v/v)) in the ratio of chloroform, methanol, acetic acid and water 40:7.5:6:1.8 (v/v) Deployment was carried out. After the second development, it was stained using 5% (w/v) ethanolic molybdenum phosphate (ethanolic molybdatophosphoric acid).

In addition, when comparing the composition of polar lipids, Faecalibacterium cancerinhibens CLCC1 and Faecalibacterium prausnitzii were found to be 1 phosphatidylglycerol (PG), 2 unidentified phospholipids (PL1, PL2), and a number of unknown lipids. (unidentified lipid, L) was confirmed. In addition, it was confirmed that CLCC1 additionally has unidentified phospholipids (PL3), which are not found in Faecalibacterium prausnitzii , and the results are shown in FIG. 3B. As can be seen from the above results, it can be seen that the polar lipid production patterns of Faecalibacterium cancerinhibens CLCC1 and Faecalibacterium prausnitzii according to the present invention are different.

Example 5. Short chain fatty acid (SCFA) analysis

In order to examine the differences in the production patterns of short-chain fatty acids between Faecalibacterium prausnitzii and Faecalibacterium cancerinhibens CLCC1 strains, an analysis was requested to the Korea Institute of Medical Sciences and analysis of the distribution of short-chain fatty acids was performed.

Specifically, Faecalibacterium prausnitzii and Faecalibacterium cancerinhibens CLCC1 strain of the present invention were cultured in RCM medium for 16 hours, and then the cells were precipitated through centrifugation (4° C., 4,000 rpm, 10 min) to collect the supernatant. The collected supernatant was filtered using a 0.22um filter and then stored refrigerated. Samples for GC-MS for SCFA analysis from the supernatant were prepared using Takeshi Furuhashi's method (Takeshi Furuhashi, et al., Analytical Biochemistry, Volume 543, 2018, Pages 51-54). Agilent's 6890 series equipment was used for chain fatty acid analysis.

Faecalibacterium cancerinhibens CLCC1 and Faecalibacterium in the known short-chain fatty acids present in the culture supernatant of another species prausnitzii Faecalibacterium (Short chain fatty acid, SCFA) Compared to analysis, for both samples SCFA was detected only butyric acid (butanoic acid). Was also detected in 7.8-fold higher butyric acid (butanoic acid) content of at Faecalibacterium cancerinhibens CLCC1 sample compared to Faecalibacterium prausnitzii sample. 3C to 3D are graphs of the GC-MS analysis results.

In addition, when viewed as a percentage in each sample was acid, Faecalibacterium cancerinhibens CLCC1 culture supernatant in 91.64 percent showed the acid, in the culture supernatant of the acid Faecalibacterium prausnitzii was 76.40%. Table 3 shows the short-chain fatty acid analysis results of the Faecalibacterium cancerinhibens CLCC1 strain, and Table 4 shows the short-chain fatty acid analysis results of Faecalibacterium prausnitzii . Other substances indicated in Tables 3 to 4 below were classified as noise signals that could not accurately identify substances because they were detected in the substance library database, but their matching rate was very low. In Table 3 below, RT means retention time.

peak RT Library/ID Matching Rate (Quality) Area Percentage (% of total, %) One 2.6965 Oxirane 58 606,058 3.132 2 3.1915 Butanoic acid (CAS); n-Butyric acid 94 17,732,337 91.635 3 3.5174 Ethane, methoxy- (CAS); Methane 50 497,085 2.569 4 4.1933 2-Pentanone, 4-hydroxy-4-methyl- (CAS) 43 515,638 2.665

peak RT Library/ID Matching rate
(Quality) Area Rate (% of Total, %) One 2.6908 2-Decanone (CAS) 38 338,372 11.491 2 3.0167 Butanoic acid (CAS); n-Butyric acid 93 2,249,717 76.401 3 4.2057 2-Pentanone, 4-hydroxy-4-methyl- (CAS) 47 356,542 12.108

Butyric acid is known to have anticancer activity, and the Faecalibacterium cancerinhibens CLCC1 strain of the present invention has a higher production of butyric acid among short-chain fatty acids compared to Faecalibacterium prausnitzii , and it can be predicted to exhibit excellent anticancer activity.

Example 6. Cancer incidence inhibition efficacy test in subcutaneous tumor model using mouse liver cancer cell line

In order to confirm the cancer incidence inhibitory ability of the Faecalibacterium cancerinhibens CLCC1 strain, cancer incidence inhibitory efficacy in a subcutaneous tumor model using a mouse liver cancer cell line was tested. Liver cancer was developed in 10 6-week-old male C57BL/6 mice through a subcutaneous tumor transplantation experiment of a liver cancer cell line (Hep55.1c), and oral administration of Faecalibacterium cancerinhibens CLCC1 strain during the development of liver cancer for 55 days led to the growth of liver cancer cells. We looked at the effect on.

Specifically, Faecalibacterium cancerinhibens CLCC1 was orally administered from 4 days before transplanting the mouse liver cancer cell line into the subcutaneous tissue of the mouse. On the 4th day of oral administration, a mouse liver cancer cell line (Hep55.1c) was made to be 2.0 x 10 ⁶ cells and transplanted into the subcutaneous tissue of the right flank.

For the subcutaneous tumor model using the mouse liver cancer cell line, Faecalibacte rium cancerinhibens CLCC1 strain was orally administered once a day for 55 days and 5 times a week. Thereafter, the size (tumor volume) of the liver cancer tumor was measured using Magnetic Resonance (MR) imaging every 7 days from the day of transplantation (day 0) to the day 55.

Faecalibacterium cancerinhibens CLCC1 strain was cultured in RCM medium, concentrated, and stored frozen to a final glycerol concentration of 15% (v/v) using PBS buffer and glycerol. And it was orally administered to mice at 200 ul (2x10 ⁸ cells) after thawing immediately before administration. As a control, 10 mice to which the CLCC1 strain was not administered were used. The control group was orally administered 200ul of PBS and glycerol (15%, v/v) solution containing no strain.

Tables 5 and 6 show the tumor size results of the control and test bacteria in the liver cancer inhibition efficacy test of the isolated strain in the subcutaneous tumor model. Table 5 shows the results of changes in the volume of the tumor relative to the control group, and Table 6 shows the results of changes in the size of the tumors in the experimental group to which the Faecalibacterium cancerinhibens CLCC1 strain was administered. In the table, SD means standard deviation.

In FIG. 4, the tumor volume according to the change of time after inducing cancer in the mouse is shown as a graph, and the changes in the size of the tumor are shown in numerical values in Tables 5 to 6 above.

As a result of the experiment, it was confirmed that the incidence and progression of liver cancer were significantly inhibited in the experimental group to which CLCC1 was administered orally compared to the control group. On average, the volume of the tumor to CLCC1 compared to the control group from the experimental group administered orally showed a 75.3% approximately, the average 32.9% lower at 19.5%, Faecalibacterium cancerinhibens CLCC1 liver cancer in the strain is administered group is delayed, the growth of tumors It was confirmed that the speed of progress was inhibited.

In addition, on the 4th day, the first tumor size measurement day after surgery, the size of the tumor in the experimental group administered with the Faecalibacterium canceerinhibens CLCC1 strain was significantly smaller than that of the control group, indicating that it had a cancer-inducing effect.

Therefore, it can be seen that the orally administered CLCC1 strain has an activity to inhibit the occurrence and progression of liver cancer.

Example 7. Inhibitory efficacy test in liver transplantation model using mouse liver cancer cell line

To investigate the effect of Faecalibacterium cancerinhibens CLCC1 strain on the growth of liver cancer cells, four 8-week-old male C57BL/6 mice developed liver cancer through a liver transplantation experiment with a liver cancer cell line, and Faecalibacterium cancerinhibens bacteria in the process of developing liver cancer for 39 days. The effect of oral administration of CLCC1 on the growth of liver cancer cells was examined.

Specifically, the liver cancer cell line Hep55.1c was used, and the epigastric part of the 8-week-old male C57BL/6 mouse was opened and transplanted to the left lobe of the liver to become 5.0 x 10 ⁵ cells. After transplantation of a tumor cell line in the liver, after a recovery period for 4 days, the size of the tumor was confirmed through MRI, and 4 well-formed tumors were selected and used in subsequent experiments.

For 39 days from the day of the transplant operation (day 0), the CLCC1 strain was prepared in the same manner as in Example 4 and orally administered to 4 mice that completed the transplant operation, and the CLCC1 strain was not administered to the 4 control mice. The control group was orally administered 200ul of PBS and glycerol (15%, v/v) solution containing no strain. Thereafter, the size and relative size of the tumor and the relative value of growth were measured periodically.

The size of the tumor was measured by magnetic resonance (MR) imaging every 7 days, and the relative value of the tumor size was derived by calculating the relative size to the tumor size on the day of transplantation. 5A to 5C show the results and graphs of the cancer-inducing inhibitory effect test results of the CLCC1 strain in the liver transplantation model.

Figure 5a is a point graph and a line graph directly comparing the size of the tumor, Figure 5b is a point and line graph expressing the size change of the tumor in fold, and Figure 5c is the amount of change in the size of the tumor (tumor size and It is a point and line graph showing the difference in tumor size on the previous measurement day).

As a result, it was confirmed that the incidence and progression of liver cancer were significantly suppressed in the experimental group administered with CLCC1 orally compared to the control group not administered with the isolate. Results of the inhibition of liver cancer and tumor size change data are shown in FIGS. 5A to 5C.

Table 7 shows the relative growth (Fold) of tumor size, and Table 7 shows the relative growth (Delative) of tumor growth.

As can be seen in Tables 7 and 8 and FIGS. 5A to 5B, it was confirmed that tumor growth of the mice to which the CLCC1 strain was administered orally was significantly suppressed compared to the control group. Therefore, it can be confirmed that the Faecalibacterium kenserinhibins CLCC1 strain has excellent tumor growth inhibitory ability.

Example 8. Confirmation of anticancer effect in mouse subcutaneous tumor model using human colon cancer cell line

8-1. Experimental method to confirm anticancer effect

In order to confirm the ability of the Faecalibacterium cancerinhibens CLCC1 strain to inhibit cancer incidence, the efficacy of inhibiting cancer incidence in a subcutaneous tumor model using a human colon cancer cell line was tested. HCT-116 cells were used as the colon cancer cell line, and immunodeficient nude mice (CAnN.Cg-Foxn1nu/CrlOri, SPF) were used as mice.

Specifically, the colon cancer cell line HCT-116 (2.5Х10 ⁷ cells/mL) was injected into mice that had undergone a one-week acclimatization period by administering 0.2 mL/head subcutaneously to the right side of the mouse using a disposable syringe.

When the size of the cancer cells grew to about 85-119 mm ³ after cancer cell transplantation, 10 mice per group were classified as a negative control group without CLCC1 administration and an experimental group 3 group receiving CLCC1 by concentration. Specifically, the test group were divided into three groups of high-low, medium, depending on the concentration of CLCC1, a low concentration is 2x10 ⁶ cells / head, jungnong degree of 2x10 ⁷ cells / head, a high concentration of F at a concentration of 2x10 ⁸ cells / head The cancerinhibens CLCC1 strain was administered. The strain was administered orally by syringe once daily for 36 days. The negative control group was administered only an excipient (a PBS solution containing 15% (v/v) glycerol).

Table 9 shows the information of the negative control group and each experimental group.

sign CLCC1 dosage Dosage Explanation Population (heads) G1 0 0.2mL/head Negative control 10 G2 2x10 ⁶ (cells/head) 0.2mL/head Low concentration experimental group 10 G3 2x10 ⁷ (cells/head) 0.2mL/head Medium concentration experimental group 10 G4 2x10 ⁸ (cells/head) 0.2mL/head High concentration experimental group 10

8-2. Tumor size change and growth inhibition rate according to strain administration

In order to confirm the anti-colorectal cancer effect of the F. cancerinhibens CLCC1 strain, the size change of the tumor was confirmed. Specifically, after the start of the strain administration test of Example 7-1, general symptoms such as appearance, behavior, and excretion were observed once a day, and the weight was measured once a week, the size of the tumor was measured twice a week, and the volume was calculated. I did. Figures 6c and 6d show the results of visual observation of the size of the tumor.

The volume (Tv) of the tumor was calculated by the method of Equation 2 below by measuring the perpendicular width (W) and the maximum length (L) of the tumor, respectively.

The results of confirming the change in tumor volume over 36 days are shown in Table 10 below and FIG. 6A. Compared to the negative control group (G1), it was confirmed that the tumor size decreased statistically significantly in all experimental groups administered with the CLCC1 strain. In Table 10 below, Mean means the average volume, and S.D. means the standard deviation.

* p<0.05, Significant difference from the negative control group (G1) by Dunnett's t-test.

** p<0.01, Significant difference from the negative control group (G1) by Dunnett's t-test.

Specifically, while the average tumor size for the current day (one day), start strain administration, but it seemed to 105mm ³ all four groups, after 36 days the average tumor volume from negative control appears to 4330mm ^3, CLCC1 strain is administered All of the experimental groups showed a volume of 3000 mm ³ or less, and it was confirmed to have a tumor volume of about 60 to 67% compared to the control group, and the effect of inhibiting the growth of colon cancer was confirmed by the Faecalibacterium cancerinhibins strain.

Also, based on the results of measuring the weight of the tumor, the tumor growth inhibition rate (IR) in each experimental group was calculated by the method of Equation 1 below.

[Equation 1]

In Equation 1, T is the average tumor weight in each experimental group, and C is the average tumor weight in the negative control group.

Tables 11 and 6b show the results of measuring tumor weights in each group after the end of the strain administration experiment, and Table 12 below shows the results of measuring average weights of mice in each group according to the experiment.

** p<0.01, Significant difference from the negative control group (G1) by Dunnett's t-test

In the negative control group (G1), the weight of the tumor extracted after necropsy was about 3.21 g, but in the experimental group, the weight of the tumor tissue was significantly lower as 1.94, 2.17 and 1.95 g at low, medium, and high concentrations, respectively. On the other hand, the total body weight in each group did not show a significant difference between the control group and the experimental group.

In the low concentration experimental group, the tumor growth inhibition rate was 39.6%, in the medium concentration experimental group, the tumor growth inhibition rate was 32.4%, and in the high concentration experimental group, the tumor growth inhibition rate was 39.3%, confirming that tumor growth was inhibited by more than 30% in all experimental groups. .

8-3. Confirmation of tumor cell necrosis and apoptosis

After 36 days of CLCC1 strain administration, autopsies were performed, and H&E staining (Hematoxylin & Eosin staining) to see necrosis of tumor cells in tumor tissue and TUNEL assay to see apotosis were performed.

Figure 6e shows a picture of the result of H&E staining for necrosis analysis of tumor cells. There was no significant difference in the level of necrosis of tumor cells between the negative control group and the test group. Therefore, it can be seen that the cell line of Pacalibacterium cancer inhibins of the present application does not induce an inflammatory response due to tumor necrosis.

Figure 6f shows the results of the TUNEL staining analysis for the analysis of apoptosis of tumor cells. It was confirmed that the apoptosis of tumor cells was statistically significantly increased in the experimental group administered with Pacalibacterium compared to the negative control group. Therefore, it was confirmed that the decrease in tumor volume in each Pacalibacterium administration group was not due to necrosis of the tumor, but due to apoptosis of the tumor and did not induce an inflammatory response.

Table 13 shows the results of comparing the necrosis and apoptosis of tumor cells. In the table below, ± is minimal, + is mild, ++ is moderate, +++ is marked, and ++++ is severe, depending on the degree of cell death. Show. The number of dead cells corresponding to the corresponding stage is indicated under each symbol.

## p<0.01, Significant difference from the negative control group (G1) by Steel's t-test.

As can be seen in Table 13, the control group (Control, G1) showed a cell death rate of about 10.33%, and the low concentration experimental group (G2) showed a cell death rate similar to that of the control group, but the medium concentration (G3) And in the high concentration (G4) experimental group, more than 12.4% apoptosis was observed. Therefore, it was confirmed that the apoptosis of tumor cells increased due to the administration of the Faecalibacterium cancerinhibis strain.

Korea Research Institute of Bioscience and Biotechnology KCTC13783BP 20190103

<110> ChunLab, Inc. <120> Faecalibacterium cancerinhibens and anti-cancer composition for colon cancer comprising the same <130> DPP20193013KR <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1499 <212> DNA <213> Artificial Sequence <220> <223> Faecalibacterium cancerinhibens CLCC1 16S rRNA sequence <400> 1 agagtttgat cctggctcag gacgaacgct ggcggcgcgc ctaacacatg caagtcgaac 60 gagcgagaga gagcttgctt tctcgagcga gtggcgaacg ggtgagtaac gcgtgaggaa 120 cctgcctcaa agagggggac aacagttgga aacgactgct aataccgcat aagcccacgg 180 ctcggcatcg agcagaggga aaaggagcaa tccgctttga gatggcctcg cgtccgatta 240 gctagttggt gaggtaatgg cccaccaagg cgacgatcgg tagccggact gagaggttga 300 acggccacat tgggactgag acacggccca gactcctacg ggaggcagca gtggggaata 360 ttgcacaatg ggggaaaccc tgatgcagcg acgccgcgtg gaggaagaag gtcttcggat 420 tgtaaactcc tgttgttggg gaagataatg acggtaccca acaaggaagt gacggctaac 480 tacgtgccag cagccgcggt aaaacgtagg tcacaagcgt tgtccggaat tactgggtgt 540 aaagggagcg caggcgggaa gacaagttgg aagtgaaatc tatgggctca acccataaac 600 tgctttcaaa actgtttttc ttgagtagtg cagaggtagg cggaattccc ggtgtagcgg 660 tggaatgcgt agatatcggg aggaacacca gtggcgaagg cggcctactg ggcaccaact 720 gacgctgagg ctcgaaagtg tgggtagcaa acaggattag ataccctggt agtccacacc 780 gtaaacgatg attactaggt gttggaggat tgaccccttc agtgccgcag ttaacacaat 840 aagtaatcca cctggggagt acgaccgcaa ggttgaaact caaaggaatt gacgggggcc 900 cgcacaagca gtggagtatg tggtttaatt cgacgcaacg cgaagaacct taccaagtct 960 tgacatccct tgacagacat agaaatatgt tttctcttcg gagcaaggag acaggtggtg 1020 catggttgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc 1080 cttatggtca gttactacgc aagaggactc tggccagact gccgttgaca aaacggagga 1140 aggtggggat gacgtcaaat catcatgccc tttatgactt gggctacaca cgtactacaa 1200 tggcgttaaa caaagagaag caagaccgcg aggtggagca aaactcagaa acaacgtccc 1260 agttcggact gcaggctgca actcgcctgc acgaagtcgg aattgctagt aatcgtggat 1320 cagcatgcca cggtgaatac gttcccgggc cttgtacaca ccgcccgtca caccatgaga 1380 gccgggggga cccgaagtcg gtagtctaac cgcaaggagg acgccgccga aggtaaaact 1440 ggtgattggg gtgaagtcgt aacaaggtag ccgtaggaga acctgcggct ggatcacct 1499

Claims (10)

Faecalibacterium spp., containing a 16S rRNA gene consisting of a nucleotide sequence having a sequence identity of 97% or more with the nucleotide sequence of SEQ ID NO: 1, a culture of the strain, and a lysate of the strain. , And a pharmaceutical composition for the prevention or treatment of colon cancer, comprising at least one active ingredient selected from the group consisting of an extract of the cells, cultures or lysates. The method of claim 1, wherein the strain is 15:1 w8c fatty acid, 19:0 cyclo w10c/19w6 fatty acid, 20:1 w9c fatty acid, 14:0 3OH/16:1 iso I fatty acid, 15:0 3OH fatty acid, 16: The composition comprising at least one fatty acid selected from the group consisting of 0 iso fatty acids and 16:0 iso 3OH fatty acids. The composition of claim 1, wherein the strain is Faecalibacterium cancer inhibins. The composition of claim 3, wherein the strain is a strain deposited with accession number KCTC 13783BP. The composition of claim 1, wherein the strain is cultured under an anaerobic condition and a temperature of 25 to 40°C. The composition of claim 1, wherein the composition has an activity of inhibiting the growth of colon cancer tumors. The composition of claim 1, wherein the composition induces apoptosis of colon cancer cells. The composition of claim 1, wherein the colon cancer is a malignant tumor occurring at one or more sites selected from the group consisting of ascending colon, transverse colon, descending colon, sigmoid colon and rectal mucosa. The composition of claim 1, wherein the colorectal cancer is one or more tumors selected from the group consisting of adenocarcinoma, lymphoma, malignant carcinoma, leiomyosarcoma, Kaposi's sarcoma, and squamous cell carcinoma. Cells of a strain of the genus Pacalibacterium, a culture of the strain, a lysate of the strain, and the above, containing a 16S rRNA gene consisting of a nucleotide sequence having a sequence identity of 97% or more with the nucleotide sequence of SEQ ID NO: 1 Food composition for preventing or improving colon cancer, comprising at least one selected from the group consisting of an extract of cells, cultures or lysates.

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