Combination therapy of lactobacillus plantarum strains and cancer treatment method using same

Abstract

The present invention relates to a pharmaceutical composition for treating cancer, in which a Lactobacillus plantarum strain and an anticancer agent are administered in combination, and the Lactobacillus plantarum strain or a culture solution of the strain not only can inhibit proliferation of cancer cells or induce apoptosis of cancer cells, but also can inhibit proliferation of cancer cells or induce apoptosis of cancer cells when administered in combination with the anticancer agent. The compound has a synergistic effect in the aspect of inhibiting proliferation of cancer cells and tumors, and can be effectively used for preventing, treating or improving cancers in the form of a pharmaceutical composition or a health-care functional food.

Description

Technical Field

[0001] The invention relates to a composition for preventing or treating cancer using a combination therapy comprising a Lactobacillus plantarum strain and an anticancer agent. Background Art

[0002] Microbiome is a compound word of microbe and genome. Human microbiome refers to the genome of all microorganisms living in the human body. Several parts of the human body are composed of various types of microorganisms. 70% of the human microbiome is distributed in the digestive tract, among which the most microorganisms live in the large intestine. It has been reported that because the microbiome is closely related to the human immune system and physical development, it is highly correlated with a variety of diseases such as obesity, metabolic diseases such as diabetes, inflammatory bowel disease, and depression. After the initial completion of the Human Genome Project in 2002, attempts were made to confirm the association between genes and intractable diseases, but it was found that environmental factors, especially the microbiome environment, may have a greater impact on the disease. Microbiome therapeutic agents, as living organisms, proliferate in the intestines and affect the human body through interactions with human cells. Therefore, as an innovative new drug, it can improve the low efficacy and high recurrence rate of existing drugs for intractable diseases, and its value has attracted much attention. In addition, with the US FDA approving the first microbiome-based treatment for Clostridium difficile infection (CDI) in December 2022, Ferring Pharmaceuticals Microbiome therapeutics have begun to be commercialized, and Seres Therapeutics' "SER-109" is also the first oral microbiome therapeutic and is currently under FDA review.

[0003] Cancer is considered one of the major health problems worldwide. Cancer is ranked as the second leading cause of death in the world. Once it develops, it requires intensive treatment and continuous management, which places a heavy burden on individuals and the socio-economic system. As of 2021, the global anticancer agent market size is about 225 trillion won. Due to the increase in the number of patients, the growth of immune anticancer agents, and the emergence of new therapies, it will grow at an average annual rate of 11% in the future, and its size is expected to reach about 470 trillion won by 2028.

[0004] Traditional cancer treatment involves a combination of chemotherapy, surgery, hormone therapy and / or radiotherapy to eliminate the patient's new cells. Cancer chemotherapy is preferably based on drugs that kill replicating cells faster than normal cells of the patient. Anticancer agents intervene in the metabolic pathways of cancer cells, directly act on DNA, block DNA replication, transcription and translation processes, or interfere with the synthesis of nucleic acid precursors, hinder cell division, and thus show cytotoxicity to cells. Therefore, these anticancer agents not only selectively act on cancer cells, but also cause fatal damage to normal cells, especially tissue cells with active cell division, leading to various serious side effects, such as vomiting, decreased bone marrow function, gastrointestinal dysfunction, hair loss, diarrhea, liver and kidney toxicity, etc. Alternatively, immune anticancer agents (or immune checkpoint inhibitors) have received great attention as third-generation anticancer agents, but their use requirements are very limited. Its therapeutic effect is usually only reflected in less than 30% of cases. Therefore, in order to completely conquer the disease of cancer, it is necessary to pay more attention to the study of the mechanism of enhancing the efficacy of immune anticancer agents of intestinal microorganisms.

[0005] In recent years, combination therapy for enhancing the effectiveness of disease treatment has been on the rise. Therefore, it is necessary to develop a combination therapy that can synergistically improve the therapeutic effect using a single component. Summary of the invention Technical issues

[0006] In one embodiment, a pharmaceutical composition for preventing or treating cancer is provided, wherein the composition comprises a first active substance and a second active substance as effective ingredients, the first active substance comprises a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and the second active substance comprises an immune anticancer agent as a first anticancer agent; or, the composition comprises the first active substance as an effective ingredient, and the second active substance is administered in combination with the first active substance.

[0007] In another embodiment, a health functional food for preventing or improving cancer is provided, wherein the health functional food contains a first active substance and a second active substance as effective ingredients, the first active substance contains a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and the second active substance contains an immune anticancer agent as a first anticancer agent; or, the health functional food contains the first active substance as an effective ingredient, and the second active substance is administered in combination with the first active substance.

[0008] In yet another embodiment, a kit for preventing or treating cancer is provided, wherein the kit comprises a first active substance and a second active substance as effective ingredients, the first active substance comprises a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and the second active substance comprises an immune anticancer agent as a first anticancer agent; or, the kit comprises the first active substance as an effective ingredient, and the second active substance is administered in combination with the first active substance.

[0009] In yet another embodiment, a method for delivering a drug into an individual is provided, the method comprising the following steps: administering a composition to an individual in need thereof, wherein the composition comprises a first active substance and a second active substance as effective ingredients, the first active substance comprises a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and the second active substance comprises an immune anticancer agent as a first anticancer agent; or, the composition comprises the first active substance as an effective ingredient, and the second active substance is administered in combination with the first active substance.

[0010] In yet another embodiment, a method for preventing or treating cancer is provided, the method comprising the following steps: administering a composition to an individual in need thereof, the composition comprising a first active substance and a second active substance as active ingredients, the first active substance comprising an effective amount of a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and the second active substance comprising an immune anticancer agent as a first anticancer agent; or, the composition comprises the first active substance as an active ingredient, and the second active substance is administered in combination with the first active substance.

[0011] Yet another embodiment provides the use of a composition in preparing a pharmaceutical preparation or a health functional food for preventing or treating cancer, wherein the composition comprises a first active substance and a second active substance as effective ingredients, the first active substance comprises a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and the second active substance comprises an immune anticancer agent as a first anticancer agent; or, the composition comprises the first active substance as an effective ingredient, and the second active substance is administered in combination with the first active substance. Technical Solution

[0012] In one embodiment, a Lactobacillus strain having anti-cancer activity, for example, anti-cancer activity against colorectal cancer is provided, specifically, Lactobacillus plantarum GB104 strain.

[0013] Lactobacillus is a class of aerobic or facultative anaerobic Gram-positive bacillus microorganisms widely distributed in nature. Microorganisms belonging to the genus Lactobacillus include Lactobacillus plantarum, Shak, etc. In order to develop new strains with excellent anti-cancer effects, the inventors screened Lactobacillus plantarum GB104 as an anti-cancer candidate strain through research. The strain was deposited in the Bioresource Center of the Korea Institute of Life Science and Technology on January 14, 2020, with the deposit number: KCTC14107BP. The strain is equivalent to a probiotic strain, which is harmless to the human body and does not produce side effects when used.

[0014] The lactobacillus has been renamed as Limosilactobacillus or Lactiplantibacillus, and the renamed strain names are used interchangeably in this specification. For example, the strain name of Lactobacillus plantarum has been renamed as Lactiplantibacillus plantarum.

[0015] In the present specification, the term "Lactobacillus plantarum GB104" can be described as L. Plantarum GB104 strain or Lactobacillus plantarum GB104 strain (Deposit No.: KCTC14107BP).

[0016] In one specific example, the strain may be a strain with a deposit number of KCTC14107BP.

[0017] In a specific example, the strain may be a strain comprising a 16S rRNA gene consisting of the nucleotide sequence of SEQ ID NO: 1.

[0018] In a specific example, the strain may be a strain having a 16S rRNA consisting of the nucleotide sequence of SEQ ID NO: 1, or a strain having a 16S rRNA containing a nucleotide sequence that is 97% or more identical to the nucleotide sequence thereof. Specifically, the strain may have at least 93%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.8%, 99.9% or 100% homology to the nucleotide sequence consisting of SEQ ID NO: 1 in the specification.

[0019] In one embodiment, the strain can be a strain that is a naturally occurring strain mutant.

[0020] In a specific example, the strain may be a live bacterium, a dead bacterium, or a cytoplasmic fraction obtained by destroying the strain, preferably a live bacterium.

[0021] In the present specification, the term "culture" can be used interchangeably with "culture supernatant", "culture supernatant", "conditioned culture fluid" or "adjusted medium", and can refer to the entire culture medium containing the strain, its metabolites, additional nutrients, etc. obtained after culturing the strain for a certain period of time in a culture medium capable of providing nutrients so that the Lactobacillus strain can grow and survive in vitro. The culture means a product obtained by culturing the probiotic strain in a well-known culture medium, and the product may or may not contain the strain itself. The culture medium can be selected from well-known liquid culture media or solid culture media, such as but not limited to MRS liquid culture medium, GAM liquid culture medium, MRS agar culture medium, GAM agar culture medium and BL agar culture medium.

[0022] In this manual, the term "lysate" can be used interchangeably with "lysate", meaning a solution or suspension of cells of a microorganism such as broken plant lactobacillus in an aqueous medium. Cell lysates include, for example, macromolecules such as DNA, RNA, proteins, peptides, carbohydrates, lipids, etc., and / or micromolecules such as amino acids, sugars, fatty acids, etc., or parts thereof. The lysate also includes cell fragments, the structure of which may be smooth or granular.

[0023] The culture broth may include the culture broth itself obtained by culturing the strain, a concentrate thereof, or a lyophilized product thereof, or a culture supernatant obtained by removing the strain from the culture broth, a concentrate thereof, or a lyophilized product thereof.

[0024] The culture solution can be obtained by culturing Lactobacillus plantarum in an appropriate culture medium (eg, MRS plate medium) at any temperature above 10° C. or below 40° C. for a certain period of time, for example, 4 to 50 hours.

[0025] In one specific example, the first active substance may have anti-cancer activity, and the first active substance comprises the strain, the culture of the strain, the lysate of the strain, or one or more selected from the following group: the bacterial body, culture and lysate extracts.

[0026] Specifically, the anticancer activity may be the activity of delaying tumor development or inhibiting tumor growth rate.

[0027] In one specific example, the first active substance may be a substance having tumor growth inhibition or tumor metastasis inhibition activity.

[0028] In one specific example, the first active substance may be a substance that induces apoptosis of cancer cells.

[0029] According to one embodiment, the anti-cancer activity can be the growth inhibition activity of cancer cells. When the supernatant of Lactobacillus plantarum GB104 strain is used to treat colorectal cancer cells HCT116, it reduces the survival rate of cancer cells by 90% to 98%; or, when Lactobacillus plantarum GB104 strain is administered to mice subcutaneously transplanted with MC-38 or HT-29 cancer cell lines, the tumor volume is 10% to 90% of the activity compared with the control group not administered with the strain.

[0030] According to one embodiment, the anti-cancer activity may be an activity of inducing apoptosis of cancer cells. When the supernatant of the plantarum Lactobacillus GB104 strain is used to treat colorectal cancer cells HCT116, the activity of inducing early apoptosis of cancer cells increases by 5 to 9 times compared with the control group not administered with the strain.

[0031] In a specific example, the first active substance may have anti-cancer activity, specifically, anti-cancer activity against colorectal cancer.

[0032] As used herein, the term "cancer" refers to a physiological condition in animals that is characterized by typically abnormal or uncontrolled cell growth. Cancer and cancer pathology may be associated with, for example, metastasis, interference with the normal function of surrounding cells, release of cytokines or other secretory products at abnormal levels, suppression or enhancement of inflammatory or immunological responses, neoplasia and premalignancy, malignancy, invasion of surrounding or distant tissues or organs, such as lymph nodes, and the like.

[0033] The cancer may be gastrointestinal cancer or non-gastrointestinal cancer.

[0034] The gastrointestinal cancer is a malignant tumor occurring in the gastrointestinal tract, such as the esophagus, stomach, small intestine or large intestine. The gastrointestinal cancer may be, for example, one or more cancers selected from the following group, but not limited to: esophageal cancer, gallbladder cancer, liver cancer, bile duct cancer, pancreatic cancer, gastric cancer, small intestine cancer, large intestine cancer, colon cancer, anal cancer and rectal cancer; in one example, it may be colorectal cancer.

[0035] The non-gastrointestinal cancer includes, but is not limited to, malignant tumors occurring in organs other than the gastrointestinal tract or the digestive system, such as, but not limited to, blood cancer, leukemia, acute myeloid leukemia, neuroblastoma, retinoblastoma, lung cancer, head and neck cancer, salivary gland cancer, melanoma, laryngeal cancer, prostate cancer, breast cancer, bladder cancer, kidney cancer, multiple myeloma, cervical cancer, thyroid cancer, ovarian cancer, urethral cancer, skin cancer, osteosarcoma, glioblastoma, brain tumor or lymphoma.

[0036] In a specific example, the cancer can be any one selected from the following group: gastric cancer, liver cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, gallbladder cancer, biliary tract cancer, cervical cancer, thyroid cancer, laryngeal cancer, acute myeloid leukemia, brain tumor, neuroblastoma, retinoblastoma, salivary gland cancer, melanoma, bladder cancer, kidney cancer, blood cancer, esophageal cancer, head and neck cancer, skin cancer, small intestine cancer, anal cancer, colon cancer, rectal cancer and lymphoma.

[0037] In one embodiment, the colorectal cancer includes one or more sites selected from the group consisting of: ascending colon, transverse colon, descending colon, sigmoid colon and rectal mucosa. The colorectal cancer may be of one or more types selected from the group consisting of, but not limited to: adenocarcinoma, lymphoma, malignant carcinoid, leiomyosarcoma, Kaposi's sarcoma and squamous cell carcinoma.

[0038] In a specific example, the first active substance may contain Lactobacillus plantarum strain alone as an effective ingredient, or may contain one or more pharmaceutically acceptable carriers, excipients or diluents.

[0040] In one specific example, the second active substance may include an anticancer agent. More specifically, the second active substance may include two different types of anticancer agents, a first anticancer agent and a second anticancer agent.

[0041] The anticancer agent may be selected from the following group: a chemical anticancer agent for chemotherapy (Chemotherapy) that can be used in combination with conventional treatments, a targeted anticancer agent, an anticancer virus, an antibody therapeutic agent, an immune anticancer agent (hereinafter interchangeably used with "immune checkpoint inhibitors"), and a combination thereof. Specifically, the anticancer agent may be a fusion protein comprising an antibody, an antigen binding fragment, or one or more single domain antibodies, or antigen binding fragments thereof, and one or more additional polypeptides. For example, the fusion protein may comprise one or more single domain antibodies and a constant region or an Fc region, and the one or more single domain antibodies, or antigen binding fragments thereof, may be non-covalently or covalently coupled to an antigen.

[0042] In one embodiment, the second active substance may include an immune anti-cancer agent.

[0043] In this specification, "antibody" means an antigen-binding fragment that can compete with a complete antibody in order to bind to any isotype of a complete immunoglobulin or a target antigen. For example, chimeric, humanized, fully humanized and bispecific antibodies or their antigen-binding fragments are included. The antibody itself is a class of antigen-binding proteins. The antibody or its antigen-binding fragment can be derived from a single source or a chimera. The chimeric antibody comprises parts derived from two different antibodies, as described in more detail below. The antibody or its antigen-binding fragment can be produced by enzymatic or chemical cleavage of a hybridoma, recombinant DNA technology or a complete antibody. Unless otherwise indicated, the term antibody in this specification may include antibodies comprising two full-length heavy chains and two full-length light chains, and derivatives, variants, fragments and mutants thereof.

[0044] In this specification, "light chain" includes a full-length light chain having a variable region sequence sufficient to provide binding specificity for an antigen or epitope and its fragment. The full-length light chain includes a variable region domain VL and a constant region domain CL. The light chain variable region domain is present at the amino terminus of the light chain polypeptide. The types of light chains include κ (kappa) and λ (lambda) chains.

[0045] In this specification, "heavy chain" includes a full-length heavy chain having a variable region sequence sufficient to provide binding specificity for an antigen or epitope and its fragments. The full-length heavy chain includes a variable region domain VH and three constant region domains CH1, CH2 and CH3. The VH domain is present at the amino terminus of the heavy chain polypeptide, the CH domain is present at the carboxyl terminus, and CH3 is located closest to the carboxyl terminus. The heavy chain includes IgG (including IgG1, IgG2, IgG3 and IgG4 subtypes), IgA (including IgA1 and IgA2 subtypes), and IgM and IgE isotypes.

[0046] The "antigen binding fragment" of the chain (heavy chain or light chain) of the antibody or immunoglobulin used in this specification comprises a part of the antibody, which lacks some amino acids compared to the full-length chain but can specifically bind to the antigen. This fragment can be considered to have biological activity in terms of being able to specifically bind to the target antigen or to compete with other antibodies or antigen binding fragments for binding to a specific epitope. On the one hand, this fragment comprises at least one CDR present in the full-length light chain or heavy chain, and in some embodiments, comprises a short chain heavy chain and / or light chain or a portion thereof. This biologically active fragment can be produced by recombinant DNA technology, for example, by enzymatic or chemical cleavage of a complete antibody. Immunologically functional immunoglobulin fragments include, but are not limited to, Fab, Fab', F(ab')2, scFab, dsFv, Fv, scFV, scFV-Fc, diabody, minibody, scAb and dAb, and can be derived from, but not limited to, any mammal, including humans, mice, rats, camelids or rabbits.

[0047] In this specification, the "Fc" region includes two heavy chain fragments containing the CH2 and CH3 domains of an antibody. The two heavy chain fragments are bound together by more than two disulfide bonds and hydrophobic interactions of the CH3 domains.

[0048] In this specification, "Fab fragment" consists of a light chain and a heavy chain containing only the variable region and CH1. The heavy chain of the Fab molecule cannot form a disulfide bond with other heavy chain molecules. ScFab is two Fab molecules connected by a flexible linker.

[0049] In the present specification, "Fab' fragment" includes the region between CH1 and CH2 domains of the heavy chain in addition to the Fab fragment, and a disulfide bond may be formed between two heavy chains of two molecules of the Fab' fragment forming the F(ab')2 molecule.

[0050] In the present specification, "F(ab')2 fragment" comprises two light chains and two heavy chains containing the variable region, CH1 and a part of the constant region between the CH1 and CH2 domains as described above, and correspondingly comprises two intrachain disulfide bonds. Therefore, the F(ab')2 fragment is composed of two Fab' fragments, and the two Fab' fragments meet through the disulfide bonds between them.

[0051] In this specification, "Fv region" is an antibody fragment that includes each variable region of the heavy chain and the light chain, but does not include the constant region. sdFV is a heavy chain and a light chain connected by a disulfide bond. scFc is a single stranded variable region (Fv) of a heavy chain and a light chain connected by a flexible linker. scFv-Fc is a connection between Fc and scFV. The mini antibody is a connection between CH3 and scFV. The diabody contains two molecules of scFV.

[0052] In the present specification, a "short-chain antibody (scAb)" is a single polypeptide chain comprising a variable region of a heavy chain and a light chain constant region connected by a flexible linker.

[0053] In the present specification, a "domain antibody (dAb)" is an immunologically functional immunoglobulin fragment containing only the heavy chain variable region or the light chain variable region.

[0054] In the present specification, a "bivalent antigen-binding protein" or "bivalent antibody" comprises two antigen-binding sites. The two antigen-binding sites contained in these bivalent antibodies may have the same antigen specificity, or may be bispecific antibodies that bind to different antigens separately.

[0055] In the present specification, a "multi-specific antigen-binding protein" or a "multi-specific antibody" means a protein that targets more than two antigens or epitopes.

[0056] In this specification, "bispecific" or "dual-specific" antigen-binding proteins or antibodies are hybrid antigen-binding proteins or antibodies with two different antigen-binding sites. These bispecific antibodies are multispecific antigen-binding proteins or multispecific antibodies and can be produced by various well-known methods, such as fusion of hybridomas, connection of Fab' fragments, etc.

[0057] In one embodiment, the additional polypeptide comprises an additional antibody or fragment thereof. The additional antibody includes, for example, complete IgG, IgE and IgM, bispecific or multispecific antibodies (e.g., etc.), single-chain Fv, polypeptide-Fc fusion, Fab, cameloid antibodies, masked antibodies (e.g., ), Small Modular ImmunoPharmaceuticals (SMIPsTM), single-chain or tandem diabodies (Tandem diabodies; ), VHH (including but not limited to those described in the present disclosure), Mini-antibodies, Ankyrin repeat protein or DART, TCR-like antibodies, MicroProteins and Additional antibodies also target PD-1, TIM-3, LAG-3, IDO, A2AR, TGFβ, CD47, or another protein involved in an immunosuppressive pathway.

[0058] In one embodiment, the additional polypeptide comprises part or all of a tumor-associated antigen (TAA) or a tumor-specific antigen (TSA), or consists thereof. Non-limiting examples of TSA or TAA antigens include differentiation antigens, such as MART-1 / MelanA (MART-I), gp100 (Pmel 17), tyrosinase, TRP-1, TRP-2, and tumor-specific multisystem antigens, such as MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, p15; overexpressed embryonic antigens, such as CEA; overexpressed tumor genes and mutated tumor-suppressor genes, such as p53, Ras, HER-2 / neu; intrinsic tumor antigens generated by chromosomal translocations, such as BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR; and viral antigens, such as Epstein-Barr virus antigen EBVA and human papillomavirus (HPV) antigens E6 and E7. Other tumor antigens include TSP-180, MAGE-4, MAGE-5, MAGE-6, RAGE, NY-ESO, erbB, p185erbB2, p180erbB-3, c-met, nm-23H1, PSA, TAG-72, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, β-catenin, CDK4, Mum-1, p15, p16, 43-9F, 5T4, 791Tgp72, alpha-fetoprotein, β-HCG, BCA225, BTAA, CA 125, CA15-3\CA27.29\BCAA, CA195, CA242, CA-50, CAM43, CD68\P1, CO-029, FGF-5, G250, Ga733\EpCAM, HTgp-175, M344, MA-50, MG7-Ag, MOV18, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, TA-90\Mac-2 binding protein\cyclophilin C-related protein, T AAL6, TAG72, TLP, MUC16, IL13Rα2, FRα, VEGFR2, Lewis Y, FAP, EphA2, CEACAM5, EGFR, CA6, CA9, GPNMB, EGP1, FOLR1, endothelial receptor, STEAP1, SLC44A4, Nectin-4, AGS-16, guanylate cyclase C, MUC-1, CFC1B, integrin α3 chain (a3b1, laminin receptor chain), and TPS.

[0059] In one embodiment, the additional polypeptide comprises part or all of a tumor antigen selected from CD19, CD20, CD22, CD30, CD72, CD180, CD171 (L1CAM), CD123, CD133, CD138, CD37, CD70, CD79a, CD79b, CD56, CD74, CD166, CD71, CLL-1 / CLECK12A, ROR1, GPC3, mesothelin CD33 / IL3Ra, c-Met, PSCA, PSMA, glycolipid F77, EGFRvIII, GD-2, MY-ESO-1 and MAGEA3, or consists of the same.

[0060] In a specific example, the immune anticancer agent may include a fusion protein, which contains a membrane protein and a cytokine involved in immune regulation. For example, the fusion protein may include a membrane protein or a fragment thereof; an Fc region and a cytokine or a variant thereof.

[0061] In this specification, the term "membrane protein" means a protein inserted into or attached to the surface of a cell membrane composed of a lipid bilayer, which transmits co-stimulatory responses and co-inhibitory responses by binding to a ligand, thereby participating in immunomodulatory proteins. In a specific example, the membrane protein may be any protein belonging to the B7 family, and more specifically, the membrane protein may be CD80 or a fragment thereof. CD80 is also known as B7-1, and CD80 is a transmembrane protein expressed on the surface of T cells, B cells, dendritic cells, and monocytes. As is well known, CD80 binds to CD28, CTLA4 (CD152), and PD-L1. CD80, CD86, CTLA4, and CD28 participate in the co-stimulation-co-inhibition system. For example, the activity of T cells is regulated, and proliferation, differentiation, and survival are involved.

[0062] In this specification, the term "cytokine" is a small protein category of about 5 to 20 kDa, which is not only important for immune cells, but also for cell signals produced by various cells. It acts through cell receptors to regulate the maturation, differentiation and activity of specific immune cell populations, thereby regulating the balance of immune responses. In a specific example, the cytokine can be an interleukin, more specifically, IL-2 or its variants. IL-2 stimulates the proliferation and differentiation of T cells, induces the production of cytotoxic T lymphocytes (cytotoxic T lymphocyte, CTL) and causes peripheral blood lymphocytes to differentiate into cytotoxic cells and lymphokine activated killer cells (lymphokine activated killer cell, LAK cell). IL-2 participates in the proliferation and differentiation of B cells, promotes the synthesis of immunoglobulins by B cells, and stimulates the production, proliferation and activation of natural killer cells (naturalkiller cell, NK cell).

[0063] In one embodiment, the immune anticancer agent can be a fusion protein comprising CD80 and IL-2 protein. More specifically, the immune anticancer agent can be a fusion protein comprising CD80 protein or a fragment thereof and IL-2 protein or a variant thereof.

[0064] The CD80 may be a full-length CD80 or a CD80 fragment. CD80 consists of 288 amino acids, and specifically, may have the amino acid sequence of SEQ ID NO: 2. The CD80 fragment may be the extracellular domain of CD80. In a specific example, the CD80 fragment may be a fragment in which the 1st to 34th amino acids from the N-terminus of the signal sequence of CD80 are excluded. Specifically, the CD80 fragment may be a protein consisting of the 35th to 242nd amino acids of SEQ ID NO: 2. In a specific example, the CD80 fragment may have the amino acid sequence of SEQ ID NO: 3.

[0065] The IL-2 may be a wild-type IL-2 or a variant thereof. The wild-type IL-2 may have an amino acid sequence of SEQ ID NO: 4. The IL-2 variant may have some amino acids substituted in the wild-type IL-2. As a specific example of an IL-2 variant with substituted amino acids, at least one of the amino acids at positions 38, 42, 45, 61, and 72 of the amino acid sequence of SEQ ID NO: 4 may be substituted.

[0066] Specifically, the IL-2 variant may have at least one substitution selected from the group consisting of R38A, F42A, Y45A, E61R and L72G in the amino acid sequence of SEQ ID NO: 4.

[0067] Preferably, a specific example of the IL-2 variant may be that any one of the following combinations (a) to (d) is substituted in the amino acid sequence of SEQ ID NO: 4:

[0068] (a)R38A / F42A

[0069] (b)R38A / F42A / Y45A

[0070] (c)R38A / F42A / E61R

[0071] (d)R38A / F42A / L72G

[0072] In one specific example, the IL-2 variant may have an amino acid sequence of SEQ ID NO:5 to SEQ ID NO:8.

[0073] The CD80 or its fragment and IL-2 or its variant can be combined through a linker or a carrier. In the present specification, linkers and carriers can also be used interchangeably. The linker connects two proteins. A specific example of a linker may include 1 to 50 amino acids, albumin or its fragment, or the Fc domain of an immunoglobulin, etc. At this time, the Fc domain of the immunoglobulin means a protein comprising the heavy chain constant region 2 (CH2) and the heavy chain constant region 3 (CH3) of an immunoglobulin, but not the variable region of the heavy chain and light chain of the immunoglobulin and the light chain constant region 1 (CH1). The immunoglobulin may be IgG, IgA, IgE, IgD or IgM, preferably, IgG4. At this time, the Fc domain of the wild-type immunoglobulin G4 may have the amino acid sequence of SEQ IDNO: 9.

[0074] Specifically, the fusion protein may have a structure in which the Fc domain is used as a linker (or carrier), and CD80 and IL-2 proteins are connected to the N-terminus and C-terminus thereof, respectively, or IL-2 and CD80 are connected. The connection between the N-terminus or C-terminus of the Fc domain and CD80 or IL-2 may be optionally achieved through a connecting peptide.

[0075] In one embodiment, the fusion protein may be composed of the following structural formula (I) or (II):

[0076] N'-X-[Linker (1)]n-Fc domain-[Linker (2)]mY-C'(I)

[0077] N'-Y-[Linker (1)]n-Fc domain-[Linker (2)]mX-C'(II)

[0078] Wherein, in the structural formula (I) and (II),

[0079] The N' is the N-terminus of the fusion protein,

[0080] The C' is the C-terminus of the fusion protein,

[0081] The X is CD80 protein or a fragment thereof,

[0082] The Y is IL-2 protein or a variant thereof,

[0083] The linker (1) and linker (2) are peptide linkers,

[0084] The n and m are independently 0 or 1.

[0085] Preferably, the fusion protein may be composed of structural formula (I). The CD80 protein or its fragment and the IL-2 protein or its variant are the same as described above.

[0086] A peptide linker (1) may be included between the CD80 protein and the Fc domain. The peptide linker (1) may be composed of 5 to 80 consecutive amino acids, 20 to 60 consecutive amino acids, or 25 to 50 consecutive amino acids, or 30 to 40 amino acids. In a specific example, the peptide linker (1) may be composed of 30 amino acids. In addition, the peptide linker (1) may contain at least one cysteine. Specifically, it may contain one, two or three cysteines. In addition, the peptide linker (1) may be derived from the hinge region of an immunoglobulin. In a specific example, the peptide linker (1) may be a peptide linker consisting of the amino acid sequence of SEQ ID NO: 10.

[0087] The peptide linker (2) may be composed of 1 to 50 consecutive amino acids, or 3 to 30 consecutive amino acids, or 5 to 15 amino acids. In a specific example, the peptide linker (2) may be (G4S)n (wherein n is an integer from 1 to 10). Wherein, n in (G4S)n may be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. In one embodiment, the peptide linker (2) may be a peptide linker consisting of the amino acid sequence of SEQ ID NO: 11.

[0088] In a specific example, the fusion protein may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 12 to SEQ ID NO: 15. More specifically, the fusion protein may have the amino acid sequence of SEQ ID NO: 12 to SEQ ID NO: 15.

[0089] In a specific example, the fusion protein can be encoded by a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the nucleic acid sequence of SEQ ID NO: 17 to SEQ ID NO: 20. More specifically, the fusion protein can have the nucleic acid sequence of SEQ ID NO: 17 to SEQ ID NO: 20.

[0090] The polynucleotide encoding the fusion protein may also include a nucleic acid encoding a signal sequence or a leader sequence. The term "signal sequence" used herein means a signal peptide that indicates secretion of the target protein. The signal peptide is translated and cut in the host cell. Specifically, the signal sequence is an amino acid sequence that starts the movement of the protein across the ER (endoplasmic reticulum) membrane. In one embodiment, the signal sequence may have the amino acid sequence of SEQ ID NO: 16.

[0091] In a specific example, the fusion protein may be a homodimer formed by binding of two identical proteins. The binding between the fusion proteins constituting the dimer may form a dimer through a disulfide bond of cysteine ​​present in the linker. More specifically, the fusion protein constituting the dimer may have a protein having an amino acid sequence of SEQ ID NO: 12 to SEQ ID NO: 14.

[0093] In one embodiment, the second active substance may further include a second anticancer agent, wherein the first anticancer agent and the second anticancer agent included in the second active substance are different from each other.

[0094] In one embodiment, the second anticancer agent can be selected from any one of the following groups: chemical anticancer agents, targeted anticancer agents and immune anticancer agents.

[0095] In this specification, the term "chemical anticancer agent" is also referred to as an antitumor drug (Antineoplastic agent) or a cytotoxic drug (Cytotoxic agent). A general term for drugs that show anticancer activity mainly by directly acting on DNA, blocking DNA replication, transcription and translation processes, or by interfering with the synthesis of nucleic acid precursors in metabolic pathways and hindering cell division. The anticancer drug not only acts on tumor cells, but also acts on normal cells and shows cytotoxicity. Chemical anticancer agents can be used for maintenance therapy (Maintenance therapy). And in this specification, the term "maintenance therapy" means a method of treating cancer with drugs after initial anticancer treatment to prevent or delay cancer recurrence.

[0096] Specifically, the chemical anticancer agent can be selected from any one of the following group: alkylating agent, microtubule inhibitor, antimetabolite and topoisomerase inhibitor. The alkylating agent can be any one selected from the following group: mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, thiotepa, altretamine, procarbazine, busulfan, streptozotocin, carmustine, lomustine, dacarbazine, cisplatin, carboplatin and oxaliplatin. The microtubule inhibitor may be selected from any one of the following group: Docetaxel, Velban, Oncovin and Navelbine. The antimetabolite may be selected from any one of the following group: 5-Fluorouracil, Capecitabine, Cytarabine, Gemcitabine, Fludarabine, Methotrexate, Pemetrexed and Mercaptopurine. The topoisomerase inhibitor may be selected from any one of the following group: Hycamtin, Camptosar, Vepesid, Paclitaxel, Blenoxane, Adriamycin and Cerubidine.

[0097] In this specification, the term "targeted anticancer agent" means a therapeutic agent that blocks signals involved in cancer growth and development by targeting specific proteins or specific gene changes that are only frequently found in cancer cells, thereby specifically killing cancer cells. It is divided into monoclonal antibodies that react outside the cell and small molecules that act inside the cell. Monoclonal antibodies are a type of anticancer agent that can block cancer cell-inducing signals transmitted outside the cell and act on initiation signals related to proliferation, apoptosis, etc., while small molecules act on complex signal transmission occurring inside the cell.

[0098] Specifically, the targeted protein can be epidermal growth factor receptor (EGFR), vascular endothelial growth factor receptor (VEGFR), CD20, CD38, RNAK-L, BTK, Bcr-abl, PDGFR / FGFR series, MEK / RAF, HER2 / Neu, ubiquitin, JAK, MAP2K, ALK, PARP, tumor growth factor β receptor (TGFβR), proteasome, Bcl-2, C-Met, VR1, VR2, VR3, c-kit, AXL, RET, Braf, DNA methyltransferase (DNMT), CDK4 / 6 and STING, etc.

[0099] The targeted anticancer agent can be selected from any one of the following groups: Olaratumab, Erlotinib, Panitumumab, Trastuzumab, Trastuzumab-Emtansine, Pertuzumab, Cetuximab, Rituximab, Bevacizumab, Axitinib, Lenvatinib, Ramucirumab, Aflibercept, Aflibercept, Obinutuzumab, Daratumumab, Denosumab, Ibrutinib, Dasatinib, Radotinib, Nilotinib, Imatinib, Bosutinib, Galunisertib, Vactosertib, Nintedanib, Sunitinib, Sorafenib ), Cabozantinib, Regorafenib, Masitinib, Semaxanib, Ceritinib, Tivozanib, Brigatinib, Vandetanib, Pazopanib, Trametinib, Temsirolimus, Dabrafenib, Dacomitinib, Afatinib, Lapatinib inib), Neratinib, Lenalidomide, Osimertinib, Ixazomib, Olmutinib, Everolimus, Ruxolitinib, Lestaurtinib, Pacritinib, Cobimetinib, Selumetinib, Binimetinib, Bortezomib, Alectinib,Crizotinib, Venetoclax, Bemcentinib, Gilteritinib, Selpercatinib, Pralsetinib, Vemurafenib, Olaparib, Talazoparib, Niraparib, Rucaparib, Azacitidine, Decitabine, Guadecitabine, Gefitinib, Abemaciclib, Ribociclib, Palbociclib, and DMXAA.

[0100] In this specification, the term "epidermal growth factor receptor (EGFR)" is a cell membrane receptor that regulates cell growth, division, survival and apoptosis. The expression of EGFR in tumor tissue increases in various cancers. It is well known that tumor tissue with increased EGFR has higher invasion, metastasis and anticancer drug resistance. EGFR inhibitors can be substances that hinder the EGFR, and in a specific example, can be Cetuximab, Trastuzumab, Pertuzumab, Gefitinib, Erlotinib or Panitumumab.

[0101] In the present specification, the term "vascular endothelial growth factor receptor (VEGFR)" is a cell membrane receptor of angiogenic factors that induce angiogenesis, and VEGFR inhibitors inhibit tumor growth and metastasis by hindering the angiogenesis. A specific example of a VEGF inhibitor or a VEGFR inhibitor may be Axitinib, Lenvatinib, Bevacizumab, Ramucirumab or Aflibercept.

[0102] In this specification, the term "CD20 (B lymphocyte antigen CD20)" is a protein expressed on the surface of B cells and is used as a target protein for treating B cell lymphoma. The CD20 targeted inhibitor can be rituximab or obinutuzumab.

[0103] In the present specification, the term "CD38 (Cluster of differentiation 38)" means a protein that acts as a signal transduction receptor in immune cells and regulates cell proliferation and apoptosis, and an inhibitor targeting the same may be Daratumumab.

[0104] In this specification, the term "RANK-L (Receptor activator of nuclear factor kappa-B ligand)" refers to a RANK receptor expressed on the surface of osteoclasts, which causes bone destruction when activated by binding to a ligand. RANK-L inhibitors are mainly used for cancer patients with bone metastasis or osteoporosis, and specifically may be Denosumab.

[0105] In the present specification, the term "BTK (Bruton's tyrosine kinase)" is an enzyme involved in B cell proliferation, and overexpression may lead to blood cancer. A specific example of a BTK targeted inhibitor may be ibrutinib.

[0106] As is known to all, in this specification, the term "Bcr-abl" means a fusion protein highly expressed in patients with chronic myeloid leukemia, inducing abnormal proliferation of blood cells. Specifically, the inhibitor of the protein may be dasatinib, nilotinib, imatinib or bosutinib.

[0107] In this specification, the term "tumor growth factor β receptor (TGFβR)" refers to the cell membrane receptor of tumor growth factor, which regulates the growth, migration, differentiation and apoptosis of epithelial cells and hematopoietic cells. The TGFβR targeted inhibitors include but are not limited to Galunisertib or Vactosertib.

[0108] As is known to all, in this specification, the term "PDGFR (Platelet derived growth factor)" means the cell membrane receptor of PDGF often expressed in cancer cells, which participates in angiogenesis and regulates cancer growth, metastasis and drug resistance. FGFR (Fibroblast growth factor receptor) means the receptor of fibroblast growth factor (FGF), which regulates a variety of biological processes, including cell growth, differentiation and migration. FGFR genes are prone to mutations, and these variants are common in breast cancer, uterine cancer, ovarian cancer, cervical cancer, etc. The inhibitor targeting PDGFR or FGFR can be Nintedanib, Sunitinib, Sorafenib, Cabozatinib, Lenvatinib, Regorafenib, Masitinib, Semaxanib, Tivozanib, Vandetanib, Axitinib or Pazopanib.

[0109] In this specification, the term "MEK / RAF" means an intracellular signal transduction mediator involved in cell proliferation, cell cycle regulation, cell survival, angiogenesis, cell migration, etc., which is overactive in cancer cells. The inhibitor targeting MEK / RAF can be trametinib or dabrafenib.

[0110] In this specification, the term "HER-2 / neu (human epidermal growth factor receptor 2)" regulates cell proliferation by activating PI3K / AkT. It is well known that it is overexpressed in metastatic breast cancer and ovarian cancer, etc., and can lead to anticancer drug resistance. The Her2 / neu targeted anticancer agent can be Trastuzumab, Afatinib, Lapatinib or Neratinib.

[0111] In this specification, the term "ubiquitin" maintains the homeostasis of cells by binding to other proteins and inducing protein degradation through the protease-proteasome (ubiquitin-proteasome system, Ubiquitin-proteasome system, UPS). Abnormal expression or activity of the UPS has been observed in a variety of tumors, and its inhibitors have shown anticancer activity. Specifically, inhibitors targeting ubiquitin or proteasomes can be lenalidomide or ixazomib.

[0112] As is known to all, in this specification, the term "JAK (Janus kinase, Janus kinase)" is an upstream protein of STAT, which is a transcription factor that regulates cell proliferation, cell survival, cell migration and immune response, and JAK inhibitors reduce cell proliferation and induce apoptosis by inhibiting the activity of STAT. The JAK targeted inhibitor can be Ruxolitinib, Lestaurtinib or Pacritinib.

[0113] In this specification, the term "MAP2K (mitogen-activated protein kinase kinase)" is an intracellular signal transduction mediator that participates in cell proliferation, cell cycle regulation, cell survival, angiogenesis, cell migration, etc. by phosphorylating MAPK, and is overactive in cancer cells. MAP2K targeted inhibitors can be cobimetinib, selumetinib, trametinib or binimetinib.

[0114] In this specification, the term "ALK (Anaplastic lymphoma kinase)" is a signal transduction mediator that promotes cell proliferation, cell migration and angiogenesis and inhibits cell apoptosis, and is overactive in various cancer tissues. The ALK targeted inhibitor can be Alectinib or Crizotinib.

[0115] In the present specification, the term "Bcl-2" is a protein that inhibits cell apoptosis and is overexpressed or overactive in various cancer tissues. The inhibitor targeting Bcl-2 may be Venetoclax.

[0116] In the present specification, the term "C-Met" is a receptor for hepatocyte growth factor (HGF), which activates signal transduction related to cell growth, formation, motility, survival, angiogenesis, etc. The C-Met targeted anticancer agent may be crizotinib or cabozantinib.

[0117] In this specification, the term "VR (Vanilloid receptor)" is also referred to as TRPV (Transient receptor potential vanilloid), which exists in the form of VR1, VR2, VR3, VR4, VR5 and VR6. It is well known that VR regulates cancer cell proliferation, apoptosis, migration, invasion and angiogenesis at every stage of cancer progression.

[0118] In this specification, the term "c-kit" is also referred to as CD117, and induces signal transduction that activates cell survival, proliferation and differentiation. c-kit is a proto-oncogene, and overexpression or mutation of the gene is associated with the onset of cancer.

[0119] In this specification, the term "AXL (Tyrosin-protein kinase receptor UFO)" is a tyrosine kinase receptor present on the cell surface, which mediates signal transduction involved in cell proliferation and survival. It is well known that it is involved in anticancer drug resistance in anticancer treatment. A specific example of an AXL-targeted anticancer agent may be Bemcentinib or Gilteritinib.

[0120] As is known to all, in this specification, the term "RET (Rearragned during transfection)" means a receptor that mediates signals involved in cell proliferation, apoptosis and survival, and mutations in RET are involved in the occurrence of cancer. The targeted inhibitor of RET may be, but is not limited to, Selpercatinib or Pralsetinib.

[0121] In the present specification, the term "Braf" means a MAPK signaling mediator involved in cell proliferation, cell cycle regulation, cell survival, angiogenesis, cell migration, etc., and gene mutations are observed in cancer cells. The inhibitor targeting Braf may be Vemurafenib.

[0122] In the present specification, the term "PARP (Poly [ADP-ribose] polymerase)" is a protein that recognizes damaged DNA in the cell nucleus and activates DNA repair-related proteins after being activated. PARP targeted inhibitors inhibit the proliferation of cancer cells by hindering the DNA repair of cancer cells. A specific example of the PARP targeted inhibitor may be Olaparib, Talazoparib, Niraparib or Rucaparib.

[0123] In this specification, the term "DNA methyltransferase (DNMT)" is an enzyme that attaches a methyl group to histones wrapped around DNA, inhibiting gene expression through this process. The DMNT targeted inhibitor exerts anti-cancer activity by hindering the hypermethylation of tumor suppressor genes and inducing the normal expression of tumor suppressor genes. A specific example of DNMT targeted inhibitors can be azacitidine, decitabine and guadecitabine.

[0124] In this specification, the term "CDK (Cyclin dependent kinase) 4 / 6" refers to a protein that promotes cell growth by regulating the cell cycle and is highly active in the occurrence and progression of various malignant tumors. CDK4 / 6 targeted inhibitors exert anti-cancer activity by hindering the cell cycle of cancer cells, inhibiting cell proliferation and inducing cell apoptosis. CDK4 / 6 targeted inhibitors can be Abemaciclib or Palbociclib.

[0125] In this specification, the term "STING (Stimulator of Interferon Genes)" is an in vivo sensor that recognizes DNA fragments from cancer cells and stimulates interferon genes to activate in vivo immune cells such as dendritic cells. The STING agonist exhibits immune synergistic effects and inhibits cancer angiogenesis. For example, the STING agonist can be CDNs, SB11285, DMXAA, etc.

[0126] In the present specification, the term "antibody therapeutic agent" means a therapeutic agent that exhibits an anti-cancer effect using an antibody that recognizes a specific protein of a cancer cell as an antigen. The antibody therapeutic agent can be selected from any one of the antibodies in the following group, but is not limited to Cetuximab, Trastuzumab, Trastuzumab-Emtansine, Rituximab, Ibritumomab, Tositumomab, Brentuximab, Ofatumumab, Obinutuzumab, Necitumumab, Bevacizumab, Ramucirumab, Nivolumab, Pembrolizumab, Atezolizumab, Durvalumab and Ipilimumab.

[0127] As is known to all, in this specification, the term "immune anticancer agent" means a substance that inhibits the activity of immune checkpoint proteins, which inhibits the differentiation, proliferation and activity of immune cells and is known to eliminate cancer cells by preventing cancer cells from escaping the immune system. The immune anticancer agent can be an antibody against any one selected from the following group: 2B4, 4-1BB (CD137), AaR, B7-H3, B7-H4, BAFFR, BTLA, CD2, CD7, CD27, CD28, CD30, CD40, CD80, CD83 ligand, CD86, CD160, CD200, CDS, CEACAM, CTLA-4, GITR, HVEM, ICAM-1, KIR, LAG-3, LAIR1, LFA-1 (CD11a / CD18), LIGHT, NKG2C, NKp80, OX40, PD-1, PD-L1, PD-L2, SLAMF7, TGFRp, TIGIT, Tim3 and VISTA. More specifically, any one of the following groups can be selected, but not limited to: anti-CTLA-4 antibody, anti-PD-1 antibody, anti-PD-L1 antibody, anti-PD-L2 antibody, anti-B7-H4 antibody, anti-HVEM antibody, anti-TIM3 antibody, anti-GAL9 antibody, anti-LAG3 antibody, anti-VISTA antibody, anti-KIR antibody, anti-BTLA antibody and anti-TIGIT antibody.In a specific example, the immunological anticancer agent can be selected from any one of the following groups, but is not limited to: Atezolizumab, Avelumab, Durvalumab, Nivolumab, Pembrolizumab, Abagovomab, Adecatumumab, Afutuzumab, Alemtuzumab, Anatumomab mafenatox), mepolizumab, blinatumomab, BMS-936559, catumaxomab, cemiplimab, epacadostat, epratuzumab, indoximod, inotuzumab, okitamostat, inotuzumab Ozogamicin, intelumumab, ipilimumab, isatuximab, lambalizumab, MED14736, MPDL3280A, obinutuzumab, ocaraituzumab, ofatumumab, olatatumab, pidilizumab, rituximab, ticilizumab, samalizumab, and tremelimumab.

[0128] In addition, the immune anticancer agent can be a fusion protein comprising a membrane protein and a cytokine, specifically, a fusion protein comprising a CD80 protein or a fragment thereof and an IL-2 protein or a variant thereof. The fusion protein of CD80 and IL-2 is the same as described above.

[0129] In this specification, the term "ADC (antibody drug conjugate)" is a therapeutic agent that chemically combines an antibody and a cytotoxic drug and exhibits a high anti-cancer effect through targeted delivery. It can be Gemtuzumab-Ozogamicin, Brentuximab-Vedotin, Trastuzumab-Emtansine, Inotuzumab-Ozogamicin, Eribulin-Mesylate, etc.

[0130] The anticancer agent can be any one selected from the following group: Cisplatin, Oxaliplatin, ALTIMA, Axitinib (VR1, 2, 3, PDGFR, c-kit), Galunisertib (TGFβRI), Lenvatinib (VR1, 2, 3), Ramucirumab (VR2), Cabozantinib (c-Met, VR2, AXL, RET), Olaparib (PARP ), Guadecitabine (DNMT), Docetaxel, Paclitaxel, Pemetrexed, Vemurafenib (Braf), Abemaciclib (CDK4 / 6), Cetuximab (EGFR), Durvalumab (PD-L1), Trastuzumab (Her2), DMXAA, and Keytruda (PD-1).

[0131] The plant lactobacillus GB104 strain can be used in combination with the anticancer agent and the anticancer vaccine, wherein the plant lactobacillus GB104 strain and the anticancer agent are the same as described above.

[0132] In this specification, the term "anti-cancer vaccine" means an active immunotherapy method that eliminates cancer cells by administering tumor-specific antigens (TSA) carried by cancer cells to cancer patients, thereby enhancing the immune function in the body by activating the immune system. Anti-cancer vaccines include DNA vaccines, peptide vaccines, cell vaccines, etc., depending on the type of antigen and the method of antigen delivery. Currently, cell vaccines and DNA vaccines are being developed as representative vaccines.

[0133] In a specific example, the plant lactobacillus GB104 strain can be used in combination with two (e.g., a first anticancer agent and a second anticancer agent) or more than two different anticancer agents. For example, the two anticancer agents (e.g., a first anticancer agent and a second anticancer agent) can be a chemical anticancer agent and a targeted anticancer agent; a chemical anticancer agent and an anticancer virus; a targeted anticancer agent and an antibody therapeutic agent; a chemical anticancer agent and a cell therapeutic agent; and a chemical anticancer agent and an immune anticancer agent. It can also be a targeted anticancer agent and an anticancer virus; a targeted anticancer agent and an antibody therapeutic agent; a targeted anticancer agent and a cell therapeutic agent; a targeted anticancer agent or an immune anticancer agent. It can also be an anticancer virus and an antibody therapeutic agent; an anticancer virus and a cell therapeutic agent; and an anticancer virus and an immune anticancer agent. It can also be an antibody therapeutic agent and a cell therapeutic agent; and an antibody therapeutic agent and an immune anticancer agent. In one embodiment of the present invention, the two anticancer agents can be an immune anticancer agent and a chemical anticancer agent; an immune anticancer agent and a targeted anticancer agent or an immune anticancer agent and an antibody therapeutic agent.

[0134] In one embodiment, the first anticancer agent administered in combination with Lactobacillus plantarum GB104 strain is a fusion protein comprising CD80 protein or a fragment thereof and IL-2 protein or a variant thereof, and the second anticancer agent can be selected from one or more of the following groups: 5-fluorouracil, anti-PD-1, Bevacizumab, Cetuximab and Regorafenib.

[0135] In one embodiment, the first anticancer agent and the second anticancer agent can be co-administered simultaneously, sequentially or in reverse order.

[0136] In this specification, the term "combination therapy" or "combined administration" or "in combination" means any form of simultaneous or concurrent treatment using at least two separate therapeutic agents. The components of the combination therapy can be administered simultaneously, sequentially or in any order. These components can be administered in different doses, different dosing frequencies or different routes according to appropriate methods.

[0137] Specifically, the combined administration can be the simultaneous administration of the first anticancer agent and the second anticancer agent, or the administration of the second anticancer agent after the administration of the first anticancer agent. The combined treatment method of the present invention can be defined as, for example, if the therapeutic effect measured by the degree of response, the rate of response, the period of disease progression or the period of survival is therapeutically superior to the therapeutic effect obtained by administering one or the remaining components of the combined treatment method at conventional doses, then a synergistic effect can be provided. For example, if its therapeutic efficacy is better than the effect obtained by using each of the above alone, the therapeutic effect of the combined treatment method is synergistic. In particular, without compromising one or more of the degree of response, the rate of response, the period of disease progression and the survival data, in particular, without compromising the duration of response, and compared with the use of conventional doses of each component, if the problematic side effects are reduced and / or less, the conventional doses of the first anticancer agent and the second anticancer agent can be reduced, then a synergistic effect is considered to exist.

[0138] In the present specification, the term "simultaneous administration" is not particularly limited, and means that the components of the combination therapy are administered substantially simultaneously, for example, as a mixture or sequentially in a close order.

[0139] In this specification, the term "administered sequentially" is not particularly limited, meaning that the components of the conjoint therapy are not administered simultaneously, but are administered successively or in batches, and there is a specific time interval between administrations. The time interval between the administrations of each component in the conjoint therapy may be the same or different, for example, may be selected from the scope of 2 minutes to 96 hours, 1 day to 7 days or 1 week, 2 weeks or 3 weeks. Typically, the time interval between administrations may be a few minutes to a few hours, for example, 2 minutes to 72 hours, 30 minutes to 24 hours, or 1 hour to 12 hours. Further examples include 24 to 96 hours, 12 to 36 hours, 8 to 24 hours and 6 to 12 hours of time interval.

[0140] In a specific example, the first anticancer agent and the second anticancer agent can be administered by independent routes. Those skilled in the art can independently administer each active ingredient with a suitable administration method and dosage. Specifically, the first anticancer agent and the second anticancer agent are administered intratumorally, intraarterially, intravenously, intravascularly, intrapleurally, intraperitoneally, intratracheally, intrathecally, intramuscularly, endoscopically, intralesionally, percutaneously, subcutaneously, locally, stereotactically, orally, directly injected or perfused. Specifically, the first anticancer agent and the second anticancer agent can be administered orally, intravenously or subcutaneously. More specifically, the first anticancer agent can be administered subcutaneously or intravenously, and the second anticancer agent can be administered intraperitoneally or intravenously, but are not limited thereto.

[0142] In one specific example, the second active substance may contain the first anticancer agent alone or the first and second anticancer agents as an active ingredient, or may contain one or more pharmaceutically acceptable carriers, excipients or diluents.

[0143] In one specific example, the anticancer agent can be prepared as a separate preparation for combined administration with the composition of the present invention. The anticancer agent can be prepared as an injection containing, but not limited to, preferably a dosage form for intratumoral, intraarterial, intravenous, intravascular, intrapleural, intraperitoneal, intratracheal, intrathecal, intramuscular, endoscopically, intralesional, percutaneous, subcutaneous, local (regionally), stereotactically, orally, direct injection or perfusion administration.

[0144] In the present specification, the term "comprising as an effective ingredient" means adding a Lactobacillus strain, a lysate of the strain, a culture solution, or an extract of its culture solution, and refers to formulations including adding various ingredients as sub-components for drug delivery and stabilization, etc.

[0145] In one embodiment, the first active substance and the second active substance can be co-administered simultaneously, sequentially or in reverse order.

[0146] The "strain", "first active substance", "second active substance", "first anticancer agent", "second anticancer agent" and "combined administration" are the same as described above.

[0147] Specifically, the combined administration can be the simultaneous administration of the plant lactobacillus strain and the anticancer agent, or the administration of the anticancer agent after the administration of the plant lactobacillus strain. Wherein, the anticancer agent can include a first anticancer agent and a second anticancer agent that are different from each other, and more specifically, the first anticancer agent can be an immune anticancer agent. The combined treatment method of the present invention can be defined as, for example, if the therapeutic effect measured by the degree of reaction, the reaction rate, the disease progression period or the survival period is better than the therapeutic effect obtained by administering one or the remaining components of the combined treatment method at a conventional dose, then a synergistic effect can be provided. For example, if its therapeutic effect is better than the effect obtained by using each of the above alone, the therapeutic effect of the combined treatment method is synergistic. In particular, without compromising one or more of the degree of reaction, the reaction rate, the disease progression period and the survival data, especially without compromising the duration of the reaction, and compared with the use of each component at a conventional dose, if the problematic side effects are reduced and / or less, the conventional doses of the plant lactobacillus strain and the anticancer agent can be reduced, then a synergistic effect is considered to exist.

[0148] In a specific example, the synergistic effect of the co-administration can be to reduce the dosage or number of administrations of existing anticancer agents. Specifically, when the plant lactobacillus GB104 strain is co-administered with an immune anticancer agent (e.g., a fusion protein comprising CD80 protein or its fragment and IL-2 protein or its variant), the same or similar anticancer activity is shown at a dose lower than the immune anticancer agent alone or at a fewer number of administrations. In one embodiment, the anticancer activity of the immune anticancer agent when the administration interval is 1 day when the immune anticancer agent is administered alone is the same or similar to the anticancer activity of the plant lactobacillus GB104 strain and the immune anticancer agent when the administration interval is 5 to 10 days (e.g., 7 days) when the immune anticancer agent is co-administered. Therefore, the co-administration of plant lactobacillus GB104 has a synergistic effect and can reduce the conventional dosage and number of immune anticancer agents.

[0149] In one specific example, when Lactobacillus plantarum GB104 strain is co-administered with an immune anticancer agent (e.g., a fusion protein comprising CD80 protein or a fragment thereof and IL-2 protein or a variant thereof) and a chemical anticancer agent (e.g., 5-FU), the same or similar anticancer activity can be exhibited at a dose lower than the immune anticancer agent or chemical anticancer agent alone or at a lesser number of administrations. In one embodiment, the anticancer activity of the immune anticancer agent or chemical anticancer agent when the administration interval is 1 day when the immune anticancer agent or chemical anticancer agent is administered alone is the same or similar to the anticancer activity of the Lactobacillus plantarum GB104 strain, the immune anticancer agent and the chemical anticancer agent when the administration interval is 5 to 10 days (e.g., 7 days) when the immune anticancer agent and the chemical anticancer agent are co-administered. Therefore, the co-administration of Lactobacillus plantarum GB104 has a synergistic effect and can reduce the conventional dose and frequency of immune anticancer agents and / or chemical anticancer agents.

[0150] In a specific example, the first active substance and the second active substance can be administered by independent routes. Those skilled in the art can independently administer each active ingredient with appropriate administration usage and dosage. Specifically, the plant lactobacillus GB104 strain and the anticancer agent are administered intratumorally, intraarterially, intravenously, intravascularly, intrapleurally, intraperitoneally, intratracheally, intrathecally, intramuscularly, endoscopically, intralesionally, transdermally, subcutaneously, locally, stereotactically, orally, directly injected or perfused. Specifically, the first active substance and the second active substance can be administered orally, intravenously or subcutaneously. More specifically, the first active substance can be administered orally, and the second active substance can be administered intravenously or subcutaneously, but is not limited to this.

[0151] In a specific example, the first active substance and the second active substance can be administered to the individual at intervals of 1 to 10 days, respectively. More specifically, the first active substance is administered to the individual at intervals of 1 to 3 days, and the second active substance is administered to the individual at intervals of 5 to 10 days. In one embodiment, the first active substance is administered to the individual at intervals of 1 day, and the second active substance is administered to the individual at intervals of 7 days.

[0153] In another embodiment, a pharmaceutical composition for preventing or treating cancer is provided, wherein the composition comprises a first active substance and a second active substance as effective ingredients, wherein the first active substance comprises a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or one or more selected from the following group: extracts of the bacteria, culture and lysate, and the second active substance comprises an immune anticancer agent as a first anticancer agent; or, the composition comprises the first active substance as an effective ingredient, and the second active substance is administered in combination with the first active substance.

[0154] The “strain”, “first active substance”, “anticancer agent”, “second active substance”, “anticancer activity” and “combined administration” are as described above.

[0155] In one embodiment, the pharmaceutical composition can be administered to mammals, including humans, by a variety of routes. The administration method can be any commonly used method, such as oral, cutaneous, intravenous, intramuscular or subcutaneous administration, preferably oral administration.

[0156] In a specific example, it includes a first oral preparation containing a first active substance, and a second oral preparation containing a second active substance, wherein the first and second oral preparations can be orally administered.

[0157] In the present specification, the term "prevention" may refer to any action of suppressing a disease state of an individual or delaying its onset by administering a pharmaceutical composition according to a regimen.

[0158] In the present specification, the term "treatment" may refer to any action that improves or favorably changes the symptoms of a disease state of an individual through the administration of a pharmaceutical composition according to a regimen.

[0159] According to a specific example, the composition may contain 0.001% to 80% by weight of the plant lactobacillus strain based on the total weight of the composition. In addition, the dosage of the plant lactobacillus strain may be 0.01 mg to 10000 mg, 0.1 mg to 1000 mg, 1 mg to 100 mg, 0.01 mg to 1000 mg, 0.01 mg to 100 mg, 0.01 mg to 10 mg, or 0.01 mg to 1 mg. The strain is included in the composition in a therapeutically effective amount or a nutritionally effective concentration, for example, the strain is 10 3 Up to 10 16 CFU / g, 10 3 Up to 10 15 CFU / g, 10 3 Up to 10 14 CFU / g, 10 3 Up to 10 13 CFU / g, 10 3 Up to 10 12 CFU / g, 10 4 Up to 10 16 CFU / g, 10 4 Up to 10 15 CFU / g, 10 4 Up to 10 14 CFU / g, 10 4 Up to 10 13 CFU / g, 10 4 Up to 10 12 CFU / g, 10 5 Up to 10 16 CFU / g, 10 5 Up to 10 15 CFU / g, 10 5 Up to 10 14 CFU / g, 10 5 Up to 10 13 CFU / g, 10 5 Up to 10 12 CFU / g, 10 6 Up to 10 13 CFU / g, 10 6 Up to 10 12 CFU / g, 10 7 Up to 10 13 CFU / g, 10 7 Up to 10 12 CFU / g, 10 8 Up to 10 13 CFU / g or 10 8 Up to 10 12CFU / g content is contained in the composition, or can be contained in the composition in the form of an equal number of live or dead bacteria culture. Specifically, for adult patients, 1×10 3 Up to 1×10 16 CFU / g of live or dead bacteria. However, for those skilled in the art, the dosage can be prescribed in a variety of ways according to factors such as formulation method, administration method, patient's age, weight, sex, pathological state, food, administration time, administration route, excretion rate and reaction sensitivity, and those skilled in the art can consider these factors and adjust the dosage appropriately. The number of administrations can be 1 time, or more than 2 times within the range of clinically acceptable side effects, and the administration site can also be administered at 1 or more than 2 places. For animals other than humans, the same per kg (body weight) dose as humans can also be used, or, for example, the dose can be administered using the amount converted by the volume ratio (e.g., average value) of the target animal to a human organ (such as the heart). The administration routes that can be used may include oral, sublingual, parenteral (e.g., subcutaneous, intramuscular, intra-arterial, intraperitoneal, intrathecal or intravenous), rectal, local (including transdermal), inhalation and injection, or insertion of an implantable device or substance. According to a specific example, the subject animal to be treated can be exemplified by humans and mammals for other purposes, specifically including humans, monkeys, mice, rats, rabbits, sheep, cattle, dogs, horses, pigs, etc. According to one embodiment, the composition comprises an inactivated dry strain, and can be administered at 1 g to 10 g, 0.5 g to 1.5 g, 2.5 g to 3.5 g, or 4.5 g to 5.5 g at a time, and can be administered 1 to 3 times a day.

[0160] In this specification, the term "therapeutically effective amount" means the amount of the anticancer agent of the method and use of the present invention or the pharmaceutical composition containing the anticancer agent for the method and use of the present invention that is used by the researcher, doctor or other clinician to derive the biological or medical response or desired therapeutic effect in the patient desired to be obtained. The therapeutically effective amount of the anticancer agent can vary according to factors such as the disease state, age, sex and weight of the individual, and the ability of the anticancer agent to derive the desired response in the individual. The therapeutically effective amount can also be the amount in which the beneficial effect on the treatment exceeds any toxic or harmful effect.

[0161] According to a specific example, the pharmaceutical composition may include a pharmaceutically acceptable carrier and / or additive. For example, it may include sterile water, physiological saline, conventional buffers (phosphoric acid, citric acid, other organic acids, etc.), stabilizers, salts, antioxidants (ascorbic acid, etc.), surfactants, suspending agents, isotonic agents, or preservatives, etc. For topical administration, it may also include a combination with organic substances such as biopolymers, inorganic substances such as hydroxyapatite, specifically, with a collagen matrix, polylactic acid polymers or copolymers, polyethylene glycol polymers or copolymers, and chemical derivatives thereof.

[0162] According to a specific example, when the pharmaceutical composition is formulated into a dosage form suitable for injection, the Lactobacillus bacteria can be dissolved or dispersed in a pharmaceutically acceptable carrier, or can be frozen in a dissolved or dispersed solution state.

[0163] According to a specific example, the pharmaceutical composition may appropriately contain suspending agents, solubilizing agents, stabilizers, isotonic agents, preservatives, anti-adsorbents, surfactants, diluents, excipients, pH regulators, analgesics, buffers, reducing agents, antioxidants, etc. according to the needs of the administration method or dosage form. Pharmaceutically acceptable carriers and preparations suitable for the present invention, including those exemplified above, are described in detail in the document [Remington's Pharmaceutical Sciences, 19th ed., 1995]. According to a specific example, the pharmaceutical composition may be formulated using pharmaceutically acceptable carriers and / or excipients according to methods that are easily implemented by ordinary technicians in the field to which the present invention belongs, so as to be prepared in unit dosage form or introduced into a multi-capacity container for preparation. At this time, the dosage form may be in the form of a solution, suspension or emulsion in an oily or aqueous medium, or in the form of a powder, granules, tablet or capsule.

[0164] The pharmaceutical composition is administered in a pharmaceutically effective amount. In this specification, the term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit / risk ratio suitable for medical treatment, and the effective dosage level depends on factors including the patient's disease type, severity, activity of the drug, sensitivity to the drug, administration time, route of administration and excretion rate, duration of treatment, drugs used simultaneously, and other factors well known in the medical field. The composition of the present invention can be administered as a single therapeutic agent or in combination with other therapeutic agents; it can be administered sequentially or simultaneously with traditional therapeutic agents; it can be administered in a single or multiple doses. Taking into account all of the above factors, it is important to achieve the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

[0165] In one embodiment, the anti-cancer activity of the first active substance may be more effective when administered in combination with the second active substance.

[0166] In one embodiment, when the plant lactobacillus GB104 strain and the chemical anticancer agent 5-fluorouracil were co-administered to a tumor animal model transplanted with a mouse colorectal cancer cell line MC-38, an enhanced anticancer effect was shown compared to the administration of the plant lactobacillus GB104 strain alone. Specifically, the effect of inhibiting the growth and progression of colorectal cancer tumors was confirmed. On average, the tumor volume of the experimental group co-administered with GB104 and the chemical anticancer agent was reduced by 10% to 90% compared with the control group.

[0167] In one specific example, when the Lactobacillus plantarum GB104 strain and a chemical anticancer agent are administered in combination, it can have the activity of reducing the volume of colorectal cancer tumors to a level below 90%, below 80%, below 70%, below 67%, 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%, based on 100% of the colorectal cancer tumor volume of the negative control group that was not administered with the strain and the anticancer agent.

[0168] In one embodiment, when the plant lactobacillus GB104 strain and the immune anticancer agent anti-PD-1 were co-administered to a tumor animal model transplanted with a mouse colorectal cancer cell line MC-38, an enhanced anticancer effect was shown compared to the administration of the plant lactobacillus GB104 strain alone. Specifically, the effect of inhibiting the growth and progression of colorectal cancer tumors was confirmed. On average, the tumor volume of the experimental group co-administered with GB104 and the immune anticancer agent was reduced by 10% to 90% compared with the control group.

[0169] In one embodiment, when the plant lactobacillus GB104 strain and the immune anticancer agent GI-101 or GI-102, which is a fusion protein containing a CD80 fragment and an IL-2 protein variant, are co-administered to a tumor animal model transplanted with a mouse colorectal cancer cell line MC-38, an enhanced anticancer effect is shown compared to the administration of the plant lactobacillus GB104 strain alone. Specifically, the effect of inhibiting the growth and progression of colorectal cancer tumors was confirmed. On average, the tumor volume of the experimental group co-administered with GB104 and the immune anticancer agent was reduced by 10% to 90% compared to the control group.

[0170] In one specific example, when the plant lactobacillus GB104 strain and an immune anticancer agent are administered in combination, it can have the activity of reducing the volume of colorectal cancer tumors to a level below 90%, below 80%, below 70%, below 67%, 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%, based on 100% of the colorectal cancer tumor volume of the negative control group that was not administered with the strain and the anticancer agent.

[0171] In one embodiment, when the plant lactobacillus GB104 strain and the targeted anticancer agent cetuximab, bevacizumab or regorafenib were co-administered to a tumor animal model transplanted with a mouse colorectal cancer cell line MC-38, an enhanced anticancer effect was shown compared to the administration of the plant lactobacillus GB104 strain alone. Specifically, the effect of inhibiting the growth and progression of colorectal cancer tumors was confirmed. On average, the tumor volume of the experimental group co-administered with GB104 and the targeted anticancer agent was reduced by 10% to 90% compared with the control group.

[0172] In one specific example, when the Lactobacillus plantarum GB104 strain and a targeted anticancer agent are administered in combination, it can have the activity of reducing the volume of colorectal cancer tumors to a level below 90%, below 80%, below 70%, below 67%, 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%, based on 100% of the colorectal cancer tumor volume of the negative control group that was not administered with the strain and the anticancer agent.

[0173] In one specific example, when the plant lactobacillus GB104 strain, an immune anticancer agent and a chemotherapeutic agent are administered in combination, it can have the activity of reducing the volume of colorectal cancer tumors to below 90%, below 80%, below 70%, below 67%, 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% based on the colorectal cancer tumor volume of the negative control group that was not administered with the strain and the anticancer agent as 100%.

[0174] In one embodiment, when the plant lactobacillus GB104 strain, the immune anticancer agent GI-101 and the chemical anticancer agent 5-FU were administered in a triple combination to a tumor animal model transplanted with a mouse colorectal cancer cell line MC-38, an enhanced anticancer effect was shown compared to the administration of the plant lactobacillus GB104 strain alone or the combined administration of the plant lactobacillus GB104 strain and the immune anticancer agent GI-101. Specifically, the effect of inhibiting the growth and progression of colorectal cancer tumors was confirmed. On average, compared with the control group, the tumor volume of the experimental group administered with the triple combination of GB104, the immune anticancer agent and the chemical anticancer agent was reduced by 10% to 90%.

[0176] In yet another embodiment, a health functional food for preventing or improving cancer is provided, wherein the health functional food comprises a first active substance, wherein the first active substance comprises a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or one or more extracts selected from the bacteria, culture and lysate, and a second active substance comprising an immune anticancer agent as a first anticancer agent is administered in combination with the first active substance.

[0177] In yet another embodiment, a food composition for preventing or ameliorating cancer is provided, wherein the food composition comprises a first active substance and a second active substance as effective ingredients, wherein the first active substance comprises a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or one or more selected from the following group: extracts of the bacteria, the culture and the lysate, and the second active substance comprises an immune anticancer agent as a first anticancer agent; or, the food composition comprises the first active substance as an effective ingredient, and the second active substance is administered in combination with the first active substance.

[0178] The "strain", "first active substance", "second active substance", "first anticancer agent", "second anticancer agent", "anticancer activity" and "combined administration" are as described above.

[0179] In one embodiment, the food composition can be administered to mammals, including humans, by various routes. The administration method can be any commonly used method, such as oral, cutaneous, intravenous, intramuscular or subcutaneous administration, preferably oral administration.

[0180] In a specific example, it includes a first oral formulation containing a first active substance, and a second oral formulation containing a second active substance, wherein the first and second oral formulations can be orally administered.

[0181] In one embodiment, the health functional food may further include a food-acceptable carrier.

[0182] In this specification, the term "food-acceptable" means showing the property of being non-toxic to cells or humans exposed to the compound.

[0183] In this specification, the term "improvement" may refer to all actions that at least reduce parameters related to the condition being treated, such as the severity of symptoms. In this case, the health functional food can be used simultaneously with or alone with a therapeutic agent before or after the onset of the disease to prevent or improve cancer.

[0184] In the health functional food, the active ingredient can be directly added to the food or used together with other foods or food ingredients, and can be appropriately used according to a conventional method. The mixing amount of the active ingredient can be appropriately determined according to its purpose of use (for prevention or improvement). Generally speaking, when preparing food or beverage, the health functional food can be specifically added in an amount of about 15% by weight or less of the raw material, more specifically about 10% by weight or less. However, in the case of long-term intake for health and hygiene purposes or health control purposes, the amount can be below the range.

[0185] The health functional food further comprises one or more of a carrier, a diluent, an excipient and an additive, and can be made into one of the following dosage forms: tablets, pills, powders, granules, powders, capsules and liquid dosage forms. The food to which the compound according to one embodiment can be added includes various foods, powders, granules, tablets, capsules, syrups, beverages, chewing gum, tea, vitamin complexes, health functional foods, etc.

[0186] Specific examples of the carrier, excipient, diluent and additive are at least one selected from the following group: lactose, glucose, sucrose, sorbitol, mannitol, erythritol, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium phosphate, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, polyvinyl pyrrolidone, methylcellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil.

[0187] In addition to containing the active ingredients, the health functional food may also contain other ingredients as necessary ingredients without any particular restrictions. For example, as with ordinary beverages, various flavoring agents or natural carbohydrates may be contained as additional ingredients. Examples of the natural carbohydrates may be monosaccharides, such as glucose, fructose, etc.; disaccharides, such as maltose, sucrose, etc.; and polysaccharides, such as conventional sugars such as dextrin and cyclodextrin; and sugar alcohols, such as xylitol, sorbitol, erythritol, etc. As flavor enhancers other than the above, natural flavor enhancers (thaumatin, stevia extract (such as rebaudioside A, glycyrrhizic acid, etc.) and synthetic flavor enhancers (saccharin, aspartame, etc.) may be advantageously used. The ratio of the natural carbohydrates may be appropriately determined by the selection of those skilled in the art.

[0188] In addition to the above, according to one embodiment, the health functional food may include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and flavor enhancers (cheese, chocolate, etc.), pectin acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. These ingredients can be used alone or in combination, and the proportions of these additives can also be appropriately selected by those skilled in the art.

[0189] The health functional food can be provided by mixing with an existing well-known health functional food for preventing or improving cancer or other existing health functional foods. The health functional food for preventing or improving cancer can be an existing well-known health functional food for preventing or improving metabolic diseases, an existing health functional food, or a newly developed health functional food.

[0190] When the health functional food contains other health functional foods having the effect of preventing or improving cancer, it is important to mix them in an amount that achieves the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

[0191] The food composition for preventing or improving cancer includes all forms such as functional food, nutritional supplement, health food and food additives, and the food composition of the type can be prepared into various forms according to conventional methods known in the art.

[0192] The composition of the present specification can be regarded as a food supplement. Food supplements, also known as dietary supplements or nutritional supplements, can also be regarded as another specific pharmaceutical product. Its purpose is to supplement the use of the diet and provide nutrients or beneficial ingredients that may not be taken in or cannot be taken in sufficient amounts in the normal diet. Most food supplements are regarded as foods, but are sometimes also regarded as drugs, natural health products or nutraceutical products. In the meaning of the present invention, food supplements include health functional foods. Food supplements are usually sold over the counter without prescription. When a food supplement is in the form of a pill or capsule, it contains the same pharmaceutical excipients used in drugs. However, food supplements can also be in the form of foods that fortify certain nutrients (e.g., infant formula). Therefore, in certain embodiments, the composition of the present invention is a food supplement.

[0193] The composition of the present invention can be administered directly, or mixed with a suitable edible liquid or solid, or can be in the form of tablets, pills, capsules, lozenges, granules, powders, suspensions, sachets, syrups, or freeze-dried in the form of a unit dose. It can also be in the form of monodoses of a freeze-dried composition, which are mixed in separate liquid containers provided together before administration.

[0194] The composition of the present invention may also be included in various edible foods and foods such as infant milk products. In this specification, the term "edible product" broadly includes any form of product that can be ingested by an animal in any form (e.g., a product that can be accepted by a sensory organ). The term "food product" is understood to be an edible product that provides nutritional support in vivo. Foods that are particularly noteworthy are food supplements and infant formulas. The food preferably includes carrier materials such as oatmeal gruel, lactic acid fermented foods, resistant starch, dietary fibers, carbohydrates, proteins, and glycosylated proteins. In certain embodiments, the bacterial cells of the present invention are homogenized with other ingredients such as cereals or powdered milk to form infant formulas.

[0196] In yet another embodiment, a kit for preventing or treating cancer is provided, the kit comprising a first active substance and a second active substance comprising an immune anticancer agent as a first anticancer agent as effective ingredients, the first active substance comprising a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or one or more selected from the following group: extracts of the bacteria, culture and lysate, and the second active substance comprising an anticancer agent; or, the kit comprises the first active substance as an effective ingredient, and the second active substance is administered in combination with the first active substance.

[0197] In yet another embodiment, a method for delivering a drug into an individual is provided, the method comprising the following steps: administering a composition to a desired individual, the composition comprising a first active substance and a second active substance comprising an immune anticancer agent as a first anticancer agent as effective ingredients, the first active substance comprising a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or one or more selected from the following group: extracts of the bacteria, culture and lysate, and the second active substance comprising an anticancer agent; or, the composition comprising the first active substance as an effective ingredient, and the second active substance being administered in combination with the first active substance.

[0198] In yet another embodiment, a method for preventing and treating cancer is provided, the method comprising the following steps: administering a composition to an individual in need thereof, the composition comprising a first active substance and a second active substance comprising an immune anticancer agent as a first anticancer agent as effective ingredients, the first active substance comprising a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or one or more selected from the following group: extracts of the bacteria, culture and lysate, and the second active substance comprising an anticancer agent; or, the composition comprising the first active substance as an effective ingredient, and the second active substance being administered in combination therewith.

[0199] The "strain", "first active substance", "second active substance", "first anticancer agent", "second anticancer agent", "anticancer activity" and "combined administration" are as described above.

[0200] The individual may be an individual suffering from cancer. Also, the individual may be a mammal, preferably, a human.

[0201] The administration route, dosage and administration frequency of the plant lactobacillus GB104 strain and the anticancer agent can be administered to the object in various methods and amounts according to the patient's condition and whether there are side effects, and those skilled in the art can select the best administration method, dosage and administration frequency within an appropriate range. In addition to the active ingredient, it can also be co-administered with other drugs (for example, the anticancer agent) or physiologically active substances known to have a therapeutic effect on cancer diseases, or can be prepared into a combined preparation form with other drugs. Effects of the Invention

[0202] The bacterial bodies of the plant lactobacillus strain or the culture fluid of the strain can not only inhibit the proliferation of cancer cells or induce apoptosis of cancer cells, but also have a synergistic effect in inhibiting the proliferation of cancer cells and tumors when administered in combination with anticancer agents. It can be effectively used to prevent, treat or improve cancer in the form of a pharmaceutical composition or a health functional food. BRIEF DESCRIPTION OF THE DRAWINGS

[0203] Figure 1 The graph shows the results of analyzing the cell survival rate after treating the human colorectal cancer cell line HCT116 with the culture supernatant of GB104 and other strains.

[0204] Figure 2 The graph shows the results of confirming the cell cycle after the human colorectal cancer cell line HCT116 was treated with the culture supernatant of GB104 and the control strain (WCFS1).

[0205] Figure 3The graph shows the apoptotic effect of cancer cells on human colorectal cancer cell line HCT116 or human breast cancer cell line MDA-MB-231 after treatment with culture supernatant of GB104 and control strain (WCFS1), respectively.

[0206] Figure 4 The graph shows the inhibitory effect of tumor formation on mouse MC-38 colorectal cancer cell line, LLC1 lung cancer cell line and 4T1 breast cancer cell line after treatment with the culture supernatant of GB104.

[0207] Figure 5 The graph shows the results of analyzing the synergistic effect by measuring tumor volume after combined administration of GB104 and 5-FU in an allogeneic MC-38 mouse colorectal cancer model.

[0208] Figure 6 Graph showing comparative analysis results by measuring tumor volume after single and combined administration of GB104 and 5-FU in an allograft MC-38 mouse colorectal cancer model.

[0209] Figure 7 A graph showing the results of analyzing the synergistic effect by measuring tumor volume after combined administration of GB104 and anti-PD-1 in an allogeneic MC-38 mouse colorectal cancer model.

[0210] Figure 8 Graph showing comparative analysis results by measuring tumor volume after single and combined administration of GB104 and anti-PD-1 in an allogeneic MC-38 mouse colorectal cancer model.

[0211] Fig. 9 The graph shows the results of analyzing the synergistic effect by measuring tumor volume after combined administration of GB104 and bevacizumab in a xenograft HT-29 mouse colorectal cancer model.

[0212] Fig.10 The graph shows the comparative analysis results by measuring tumor volume after single and combined administration of GB104 and Bevacizumab in a xenograft HT-29 mouse colorectal cancer model.

[0213] Fig.11 The graph shows the results of analyzing the synergistic effect by measuring tumor volume after combined administration of GB104 and cetuximab in a xenograft HT-29 mouse colorectal cancer model.

[0214] Fig.12 The graph shows the comparative analysis results by measuring tumor volume after single and combined administration of GB104 and Cetuximab in a xenograft HT-29 mouse colorectal cancer model.

[0215] Fig.13 The graph shows the results of analyzing the synergistic effect by measuring tumor volume and size after combined administration of GB104 and Regorafenib in the allogeneic MC-38 mouse colorectal cancer model.

[0216] Fig.14 The graph shows the comparative analysis results by measuring tumor volume after single and combined administration of Regorafenib in the allogeneic MC-38 mouse colorectal cancer model.

[0217] Fig.15 The graph shows the results of analyzing the synergistic effect by measuring tumor volume and weight after combined administration of GB104 and GI-101 in the allogeneic MC-38 mouse colorectal cancer model.

[0218] Fig.16 Graph showing comparative analysis results by measuring tumor volume after single and combined administration of GB104 and GI-101 in an allograft MC-38 mouse colorectal cancer model.

[0219] Fig.17 The graph shows the results of analyzing the synergistic effect by measuring tumor volume and weight after combined administration of GB104 and GI-102 in the allogeneic MC-38 mouse colorectal cancer model.

[0220] Fig.18 Graph showing comparative analysis results by measuring tumor volume after single and combined administration of GB104 and GI-102 in an allograft MC-38 mouse colorectal cancer model.

[0221] Fig.19 The graph shows the results of analyzing the synergistic effect by measuring tumor volume after triple administration of GB104, GI-101 and 5-FU in an allogeneic MC-38 mouse colorectal cancer model.

[0222] Fig. 20 The graph shows the comparative analysis results of measuring tumor volume after single and combined administration of GB104, GI-101 and 5-FU in an allogeneic MC-38 mouse colorectal cancer model. DETAILED DESCRIPTION

[0223] Preferred embodiments are given below to help understand the present invention. However, the embodiments provided below are only for easier understanding of the present invention, rather than limiting the content of the present invention. The embodiments can be variously changed, and the embodiments are not limited to the embodiments disclosed below, and can also be implemented in various forms.

[0225] Example 1 Isolation and Identification of Lactobacillus plantarum GB104 Strain

[0226] The isolation and identification of Lactobacillus plantarum GB104 strain was performed by the methods described in Korean Patent Application No. 10-2020-0186738 and Korean Patent Application No. 10-2022-0080567. The entire contents of the above documents are incorporated herein by reference.

[0227] In short, it was isolated from vaginal samples of healthy women who came to the hospital for the purpose of health examination of Lactobacillus plantarum GB104. First, a sample from the vagina was collected with a swab and streaked into Rogosa SL (MRS) plate culture medium and cultured in an anaerobic chamber at 37°C for 48 hours. After the bacterial colony grew, the single colony was subcultured on a new MRS plate culture medium for pure isolation. After pure isolation, the strain was cultured using MRS culture medium. Then, among the cultured strains, the Lactobacillus plantarum GB104 strain, which has an inhibitory effect on fat cell accumulation and has low cytotoxicity, was finally screened out. In order to identify the finally screened Lactobacillus plantarum GB104 strain, the 16S rRNA gene sequence was obtained by PCR using primers targeting the 16S rRNA gene, and analyzed by Sanger sequencing method. The 16S rRNA sequence of Lactobacillus plantarum GB104 is shown in SEQ ID NO: 1. The inventor named GB104 bacteria "Lactobacillus plantarum GB104" (deposit number: KCTC14107BP), and deposited it in the Korean collection for type cultures (KCTC) at the Korea Institute of Biotechnology on January 14, 2020. In addition, the strain name of Lactobacillus plantarum has been renamed as Lactobacillus plantarum. In the following examples, the renamed strain names of the existing strains are interchangeably described.

[0229] Experimental Example 1 Confirmation of the survival rate and apoptosis effect of cancer cells treated with GB104 strain culture supernatant

[0230] 1.1. Confirmation of the survival rate of cancer cells treated with GB104 strain culture supernatant

[0231] In this experimental example, human colorectal cancer cell lines were used to treat various L. Plantarum culture supernatants including the L. Plantarum GB104 strain, and the cell survival rate was screened by MTT analysis.

[0232] Human colorectal cancer cell line HCT116 cells were cultured at 2×10 3The cells were dispensed into each well of a 96-well plate and cultured for 24 hours. The culture supernatant of each L. Plantarum strain was treated with a concentration of 10% and cultured for 72 hours at 37°C and 5% CO2. The culture supernatant was obtained by culturing the L. Plantarum strain in an MRS medium, precipitating the strain by centrifugation, collecting only the supernatant, and filtering with a 0.22 μm filter. Using Cell Proliferation Kit I (MTT) (Roche), the MTT solution was treated to 0.5 mg / mL, and then incubated for another 4 hours to dissolve the purple formazan crystals produced by metabolic activity in the living cells with a solubilization solution, and the absorbance was measured at 570 nm, as shown in FIG. Figure 1 shown.

[0233] Figure 1 The graph shows the results of analyzing the cell survival rate after treating the human colorectal cancer cell line HCT116 with the culture supernatant of GB104 and other strains.

[0234] like Figure 1 As shown, the same cancer cell growth inhibitory effect was not shown in various L. Plantarum culture supernatants, and in particular, the GB104 culture supernatant showed an effect of reducing the cancer cell survival rate by about 95%, confirming that it is the most effective in inhibiting cancer cell growth.

[0236] Confirmation of cell viability of human cancer cell lines treated with GB104 strain culture supernatant

[0237] In this experimental example, various human cancer cell lines were used to treat the culture supernatant of L. Plantarum GB104 strain, and the cell survival rate was screened by MTT analysis.

[0238] Human cancer cell lines were cultured at 0.8-1.5×10 4 The cells were dispensed into each well of a 96-well plate and cultured for 24 hours. The supernatant of the culture of the strain L. Plantarum GB104 was treated at a concentration of 10% in the colon cancer, lung cancer, gastric cancer, breast cancer, and liver cancer cell lines, and at a concentration of 20% in the kidney cancer and bladder cancer cell lines. The cells were cultured at 37°C and 5% CO2 for 48 hours, and the cell survival rate was measured using the same method as in the above-mentioned Experimental Example 1.1. The results are shown in Tables 1 and 2.

[0239] Table 1

[0240] Table 2

[0241] The results showed that although cell survival rates varied depending on the characteristics of the cancer cells, cell survival rates of all human cancer cell lines treated with L. Plantarum GB104 culture supernatant were reduced compared to the untreated control (MRS).

[0243] Confirmation of cancer cell cycle treated with GB104 strain culture supernatant

[0244] In this experimental example, human colorectal cancer cell line was used to treat L. Plantarum GB104 culture supernatant, and the changes in the cancer cell cycle were confirmed by flow cytometry.

[0245] Human colorectal cancer cell line HCT116 cells were cultured at 5×10 4 The cells were dispensed into each well of a 6-well plate, and after culturing for 24 hours, the culture supernatant of the L. Plantarum GB104 strain was treated with a concentration of 10%, and cultured at 37°C, 5% CO2 for 48 hours. The culture supernatant was obtained by culturing the L. Plantarum strain in MRS medium, precipitating the strain by centrifugation and collecting only the supernatant, and then filtering with a 0.22 μm filter to obtain the supernatant. After 48 hours, the cells were treated with trypsin-EDTA to separate the cells, and then the cells were harvested by centrifugation, fixed with 70% ethanol, and stored at 4°C for more than 1 hour. After treating with RNase A and staining with PI (Propidium Iodine), a flow cytometer (BD FACSymphony A3 CellAnalyzer) was used to confirm the degree of apoptosis of cancer cells by DNA content analysis, such as Figure 2 shown.

[0246] Figure 2 The graph shows the results of confirming the cell cycle after the human colorectal cancer cell line HCT116 was treated with the culture supernatant of GB104 and the control strain (WCFS1).

[0247] like Figure 2 As shown, the sub-G1 region of cells treated with L. Plantarum GB104 culture supernatant increased by about 9.8 times compared with the untreated control group (MRS), thus confirming that GB104 induces apoptosis in cancer cells.

[0249] Effects of GB104 culture supernatant on early and late cancer cell apoptosis in human cancer cell lines

[0250] In this experimental example, the culture supernatant of L. Plantarum GB104 was treated with human cancer cell lines, and the apoptotic effect of cancer cells was confirmed by flow cytometry.

[0251] Human cancer cell lines were cultured at 0.5-1×10 5 The cells were dispensed into each well of a 6-well plate and cultured for 24 hours. The culture supernatant of the L. Plantarum GB104 strain was treated with a concentration of 10% and cultured for 24 hours at 37°C and 5% CO2. The culture supernatant was obtained by culturing the L. Plantarum strain in MRS medium, precipitating the strain by centrifugation and collecting only the supernatant, and then filtering with a 0.22 μm filter. After 24 hours, the cells were separated by treatment with trypsin-EDTA, and then harvested by centrifugation. After mixing the cells with 100 μL of a solution using a FITC Annexin V Apoptosis Detection Kit with 7-AAD (BioLegend), 5 μL of FITC Annexin V and 5 μL of 7-AAD were added, and the reaction was carried out at room temperature in the dark for 15 minutes. Fluorescence measurement was confirmed using a flow cytometer (BDFACSymphony A3 cell analyzer) to confirm cancer cell apoptosis, and the results are as shown. Figure 3 As shown. Normal surviving cells are not labeled with Annexin V and 7-AAD, while cells in the early stage of apoptosis are labeled with Annexin V but not with 7-AAD. Cells in the late stage of apoptosis can be distinguished and confirmed by simultaneous labeling with Annexin V and 7-AAD.

[0252] Figure 3 The graph shows the apoptotic effect of cancer cells on human colorectal cancer cell line HCT116 or human breast cancer cell line MDA-MB-231 after treatment with culture supernatant of GB104 and control strain (WCFS1), respectively.

[0253] like Figure 3 As shown, it was confirmed that the early and late apoptosis of colorectal cancer and breast cancer cell lines treated with L. Plantarum GB104 culture supernatant was significantly increased compared with the untreated control group (MRS).

[0254] This confirmed that GB104 is involved in inducing apoptosis of cancer cells.

[0256] Experimental Example 2 Tumor formation inhibition effect treated with GB104 culture supernatant

[0257] In this example, the tumor formation inhibitory effect of L. plantarum GB104 strain treatment in various mouse cancer cell lines was confirmed.

[0258] The culture supernatant was obtained by culturing the L. Plantarum strain in MRS medium, centrifuging the strain and collecting only the supernatant, and filtering with a 0.22 μm filter. Specifically, mouse colorectal cancer cell line MC-38, mouse lung cancer cell line LLC1, and mouse breast cancer cell line 4T1 were diluted to 0.5-2×10 in 2 mL of culture medium in a 6-well plate. 3 cells and dispensed into each well, and allowed to stabilize for 24 hours. After 24 hours, the culture medium was removed and replaced with a new one, and at the same time, the culture supernatant of the L. Plantarum GB104 strain was treated at a concentration of 1%. Every 2-3 days, the culture medium was replaced with a new one containing the culture supernatant of the strain and cultured for 7 days. The cultured plate was washed twice with DPBS and fixed with 4% formalin. The fixed cells were washed twice with DPBS, stained with 0.5% crystal violet solution for 5 minutes, and then washed with distilled water to confirm the colony-forming ability. The results are as follows: Figure 4 shown.

[0259] Figure 4 The graph shows the inhibitory effect of tumor formation on mouse MC-38 colorectal cancer cell line, LLC1 lung cancer cell line and 4T1 breast cancer cell line after treatment with the culture supernatant of GB104.

[0260] like Figure 4 As shown, although the tumor formation inhibitory ability varied depending on the characteristics of the cancer cells, all mouse cancer cell lines treated with L. Plantarum GB104 culture supernatant were able to inhibit tumor formation compared with the untreated control group (MRS).

[0262] Experimental Example 3 Confirmation of the inhibitory effect of GB104 strain and chemical anticancer agent 5-fluorouracil on tumor growth

[0263] In this experimental example, the tumor therapeutic effect of L. Plantarum GB104 combined with 5-fluorouracil (5-FU) was confirmed in the MC-38 allogeneic colorectal cancer model. Six-week-old c57BL / 6 mice were received, and after a one-week adaptation period, the hair around the right flank was removed, and the experiment was performed at 7 weeks of age. The colorectal cancer cell line MC-38 derived from c57BL / 6 was transplanted at 2×10 5The tumor model was established by subcutaneously injecting cells / 100 μL into the right flank of mice. The tumor size was measured using a digital calliper and calculated using the following formula: Tumor volume (mm 3 )=(width 2 x length) / 2{tumor volume (mm 3 )=(width 2 × length) / 2}. On the 5th day after tumor cell injection, only mice with tumor sizes within a certain range were selected for classification so that the average tumor size of each group was the same. Then, from the 6th day until the end of the experiment, the L. Plantarum GB104 strain was inoculated at 1×10 9 The dose of CFU was orally administered to the animal model. The chemical anticancer agent 5-FU (10 mg / kg) was intraperitoneally administered alone or in combination with GB104 once a week.

[0264] Figure 5 The graph shows the results of analyzing the synergistic effect by measuring tumor volume after combined administration of GB104 and 5-FU in an allogeneic MC-38 mouse colorectal cancer model.

[0265] Figure 6 Graph showing comparative analysis results by measuring tumor volume after single and combined administration of GB104 and 5-FU in an allograft MC-38 mouse colorectal cancer model.

[0266] like Figure 5 and Figure 6 As shown, it was confirmed that the tumor growth rate was greatly inhibited in the L. Plantarum GB104-administered group compared with the negative control group (Control). In addition, in the group administered with L. Plantarum GB104 and 5-FU alone, a significant reduction effect of tumor size due to the synergistic phenomenon was confirmed.

[0268] Experimental Example 4 Confirmation of the inhibitory effect of GB104 strain and immune anticancer agent anti-PD-1 on tumor growth

[0269] In this experimental example, the tumor therapeutic effect of L. Plantarum GB104 strain combined with anti-PD-1 was confirmed in the MC-38 colorectal cancer allograft model. Six-week-old c57BL / 6 mice were received, and after a one-week adaptation period, the hair around the right flank was removed, and the experiment was performed at 7 weeks of age. The colorectal cancer cell line MC-38 derived from c57BL / 6 was transplanted at 2×10 5The tumor model was established by subcutaneously injecting 100 μL of cells into the right flank of mice. The tumor size was measured using a digital calliper and calculated using the following formula: tumor volume (mm 3 )=(width 2 × length) / 2. On the 5th day after tumor cell injection, only tumors with a size of 10-30 mm were selected. 3 The mice were randomly divided into groups and then treated with L. Plantarum GB104 at a rate of 1 × 10 per mouse per day from day 5 until the end of the experiment. 9 The dose of CFU was orally administered to the animal model. The immune anticancer agent anti-PD-1 (5 mg / kg) was intraperitoneally administered alone or in combination with GB104 twice a week.

[0270] Figure 7 A graph showing the results of analyzing the synergistic effect by measuring tumor volume after combined administration of GB104 and anti-PD-1 in an allogeneic MC-38 mouse colorectal cancer model.

[0271] Figure 8 Graph showing comparative analysis results by measuring tumor volume after single and combined administration of GB104 and anti-PD-1 in an allogeneic MC-38 mouse colorectal cancer model.

[0272] like Figure 7 and Figure 8 As shown, compared with the negative control group (Control), it was confirmed that the tumor size in the group administered with L. Plantarum GB104 and anti-PD-1 in combination had a significant reduction effect due to the synergistic phenomenon compared with the group administered with Plantarum GB104 or anti-PD-1 alone.

[0274] Experimental Example 5 Confirmation of the inhibitory effect of GB104 strain and targeted anticancer agent Bevacizumab on tumor growth

[0275] In this experimental example, the tumor therapeutic effect of L. Plantarum GB104 combined with bevacizumab (anti-VEGF) was confirmed in a xenograft model using immunodeficient mice. 6-week-old NOG mice were received, and after a 1-week adaptation period, the hair around the right flank was removed, and the experiment was performed at 7 weeks of age. The human colorectal cancer cell line HT-29 was transplanted at 3×10 per mouse. 6 The tumor model was established by subcutaneously injecting 100 μL of cells into the right flank of mice. The tumor size was measured using a digital calliper and calculated using the following formula: tumor volume (mm 3)=(width 2 × length) / 2. On the third day after tumor cell injection, only mice with tumor sizes within a certain range were selected for classification so that the average tumor size of each group was the same. Then, from the fourth day until the end of the experiment, L. Plantarum GB104 strain was inoculated at 1×10 per mouse per day. 9 The dose of CFU was orally administered to the animal model. The targeted anticancer agent bevacizumab (10 mg / kg) was intraperitoneally administered alone or in combination with GB104 once a week.

[0276] Fig. 9 The graph shows the results of analyzing the synergistic effect by measuring tumor volume after combined administration of GB104 and bevacizumab in a xenograft HT-29 mouse colorectal cancer model.

[0277] Fig.10 Graph showing comparative analysis results by measuring tumor volume after single and combined administration of GB104 and bevacizumab in a xenograft HT-29 mouse colorectal cancer model.

[0278] like Fig. 9 and Fig.10 As shown, it was confirmed that the tumor growth rate was suppressed in the L. Plantarum GB104-administered group compared with the negative control group (Control). In addition, in the group administered with L. Plantarum GB104 and bevacizumab alone, a significant reduction effect of tumor size due to the synergistic phenomenon was confirmed.

[0280] Experimental Example 6 Confirmation of the inhibitory effect of GB104 strain and targeted anticancer agent Cetuximab on tumor growth

[0281] In this experimental example, the tumor therapeutic effect of L. Plantarum GB104 combined with cetuximab (anti-EGFR) was confirmed in a xenograft model using immunodeficient mice. Six-week-old NSG mice were received, and after a one-week adaptation period, the hair around the right flank was removed, and the experiment was performed at 7 weeks of age. The human colorectal cancer cell line HT-29 was transplanted at 3×10 6 The tumor model was established by subcutaneously injecting 100 μL of cells into the right flank of mice. The tumor size was measured using a digital calliper and calculated using the following formula: tumor volume (mm 3 )=(width 2× length) / 2. On the third day after tumor cell injection, only mice with tumor sizes within a certain range were selected for classification so that the average tumor size of each group was the same. Then, from the fourth day until the end of the experiment, the L. Plantarum GB104 strain was inoculated at 1×10 9 The dose of CFU was orally administered to the animal model. The targeted anticancer agent cetuximab (40 mg / kg) was intraperitoneally administered twice a week alone or in combination with GB104.

[0282] Fig.11 The graph shows the results of analyzing the synergistic effect by measuring tumor volume after combined administration of GB104 and cetuximab in a xenograft HT-29 mouse colorectal cancer model.

[0283] Fig.12 Graph showing comparative analysis results by measuring tumor volume after single and combined administration of GB104 and cetuximab in a xenograft HT-29 mouse colorectal cancer model.

[0284] like Fig.11 and Fig.12 As shown, a significant tumor growth inhibitory effect was confirmed in the L. Plantarum GB104-administered group compared to the negative control group (Control). In addition, a tumor size reduction effect due to a synergistic phenomenon was confirmed in the L. Plantarum GB104 and cetuximab-combined administration group compared to the cetuximab-administered group alone.

[0286] Experimental Example 7 Confirmation of the inhibitory effect of GB104 strain and targeted anticancer agent Regorafenib on tumor growth

[0287] In this experimental example, the tumor therapeutic effect of L. Plantarum GB104 combined with Regorafenib was confirmed in the MC-38 colorectal cancer allograft model. Five-week-old c57BL / 6 mice were received, and after a one-week adaptation period, the hair around the right flank was removed, and the experiment was performed at 6 weeks of age. The colorectal cancer cell line MC-38 derived from c57BL / 6 was transplanted at 2×10 5 The tumor model was established by subcutaneously injecting 100 μL of cells into the right flank of mice. The tumor size was measured using a digital calliper and calculated using the following formula: tumor volume (mm 3 )=(width 2× length) / 2. On the 5th day after tumor cell injection, only mice with tumor sizes within a certain range were selected for classification so that the average tumor size of each group was the same. Then, from the 5th day until the end of the experiment, the L. Plantarum GB104 strain was inoculated at 1×10 9 The dose of CFU was orally administered to the animal model. The targeted anticancer agent Regorafenib (3 mg / kg) was orally administered alone or in combination with GB104 every other day from day 5 until the end of the experiment.

[0288] Fig.13 The graph shows the results of analyzing the synergistic effect by measuring tumor volume and size after combined administration of GB104 and Regorafenib in the allogeneic MC-38 mouse colorectal cancer model.

[0289] Fig.14 The graph shows the comparative analysis results by measuring tumor volume after single and combined administration of Regorafenib in the allogeneic MC-38 mouse colorectal cancer model.

[0290] like Fig.13 and Fig.14 As shown, it was confirmed that the tumor growth rate was greatly suppressed in the L. Plantarum GB104-administered group compared with the negative control group (Control). In addition, in the group administered with L. Plantarum GB104 and Regorafenib alone, a significant inhibitory effect on tumor size due to the synergistic phenomenon was confirmed.

[0292] Experimental Example 8 Confirmation of the inhibitory effect on tumor growth by combined administration of GB104 strain and immune anticancer agent GI-101

[0293] In this experimental example, the tumor therapeutic effect of L. Plantarum GB104 strain and GI-101 in the MC-38 allogeneic colorectal cancer model was confirmed. Five-week-old c57BL / 6 mice were received, and after a one-week adaptation period, the hair around the right flank was removed, and the experiment was performed at 6 weeks of age. The colorectal cancer cell line MC-38 derived from c57BL / 6 was transplanted at 2×10 5 The tumor model was established by subcutaneously injecting 100 μL of cells into the right flank of mice. The tumor size was measured using a digital calliper and calculated using the following formula: tumor volume (mm 3 )=(width2 × length) / 2. On the 5th day after tumor cell injection, only tumors with a size of 10-30 mm were selected. 3 The mice were randomly divided into groups and then treated with L. Plantarum GB104 at a rate of 1 × 10 per mouse per day from day 5 until the end of the experiment. 9 The dose of CFU was orally administered to the animal model. The immunological anticancer agent GI-101 was subcutaneously injected at 3 mg / kg once a week from day 5 alone or in combination with GB104.

[0294] The immune anticancer agent GI-101 was prepared with reference to the contents disclosed in Korean Patent Publication No. 10-2020-0032009. Specifically, in order to produce a fusion protein comprising a human CD80 fragment, an Fc domain and an IL-2 variant, a polynucleotide comprising a base sequence encoding the fusion protein (SEQ ID NO: 17) was loaded into a pCGS3 vector using the BioXPTM 3250 system. The base sequence encoding the fusion protein comprises, from the N-terminus, a signal peptide (SEQ ID NO: 16), a CD80 fragment (SEQ ID NO: 3), an Ig hinge (SEQ ID NO: 10), an Fc domain (SEQ ID NO: 9), a linker (SEQ ID NO: 11), and an IL-2 variant (2M) (R38A, F42A) (SEQ ID NO: 5) in which two amino acids are substituted. The vector is introduced into CHO cells (Expi-CHO TM , Thermo Fisher Scientific) to express the fusion protein of SEQ ID NO: 12. After the vector was introduced, the culture medium was collected and the fusion protein was purified after culturing at 37°C, 125RPM, 8% CO2 concentration for 7 days.

[0295] Fig.15 The graph shows the results of analyzing the synergistic effect by measuring tumor volume and weight after combined administration of GB104 and GI-101 in the allogeneic MC-38 mouse colorectal cancer model.

[0296] Fig.16 Graph showing comparative analysis results by measuring tumor volume after single and combined administration of GB104 and GI-101 in an allograft MC-38 mouse colorectal cancer model.

[0297] like Fig.15 and Fig.16As shown, compared with the negative control group (Control), the tumor size in the L. Plantarum GB104 and GI-101 combined administration group was significantly reduced compared with the L. Plantarum GB104 or GI-101 single administration group, confirming that the antitumor efficacy was enhanced.

[0299] Experimental Example 9 Confirmation of the inhibitory effect of GB104 strain and immune anticancer agent GI-102 on tumor growth

[0300] In this experimental example, the tumor therapeutic effect of the combined administration of L. Plantarum GB104 strain and GI-102 in the MC-38 colorectal cancer allograft model was confirmed. Five-week-old c57BL / 6 mice were received, and after a one-week adaptation period, the hair around the right flank was removed, and the experiment was performed at 6 weeks of age. The colorectal cancer cell line MC-38 derived from c57BL / 6 was transplanted at 2×10 5 The tumor model was established by subcutaneously injecting 100 μL of cells into the right flank of mice. The tumor size was measured using a digital calliper and calculated using the following formula: tumor volume (mm 3 )=(width 2 × length) / 2. On the 5th day after tumor cell injection, only tumors with a size of 10-30 mm were selected. 3 The mice were randomly divided into groups and then treated with L. Plantarum GB104 at a rate of 1 × 10 per mouse per day from day 5 until the end of the experiment. 9 The dose of CFU was orally administered to the animal model. The immunological anticancer agent GI-102 was subcutaneously injected at 3 mg / kg on days 5 and 15 alone or in combination with GB104.

[0301] The immune anticancer agent GI-102 is prepared with reference to the contents disclosed in Korean Patent Publication No. 10-2020-0032009. Specifically, in order to produce a fusion protein comprising a human CD80 fragment, an Fc domain, and an IL-2 variant with three amino acids substituted, a polynucleotide comprising a base sequence encoding the fusion protein (SEQ ID NO: 18) was loaded into a pCGS3 vector using the BioXPTM 3250 system. The base sequence encoding the fusion protein comprises, from the N-terminus, a signal peptide (SEQ ID NO: 16), a CD80 fragment (SEQ ID NO: 3), an Ig hinge (SEQ ID NO: 10), an Fc domain (SEQ ID NO: 9), a linker (SEQ ID NO: 11), and an IL-2 variant (3M) (R38A, F42A, E61R) (SEQ ID NO: 7) in which three amino acids are substituted. The vector is introduced into CHO cells (Expi-CHO TM , Thermo Fisher Scientific) to express the fusion protein of SEQ ID NO: 14. After the vector was introduced, the cells were incubated at 37°C, 125 RPM, 8% CO 2 After culturing for 7 days under the same concentration, the culture medium was collected and the fusion protein was purified.

[0302] Fig.17 The graph shows the results of analyzing the synergistic effect by measuring tumor volume and weight after combined administration of GB104 and GI-102 in the allogeneic MC-38 mouse colorectal cancer model.

[0303] Fig.18 Graph showing comparative analysis results by measuring tumor volume after single and combined administration of GB104 and GI-102 in an allograft MC-38 mouse colorectal cancer model.

[0304] like Fig.17 and Fig.18 As shown, it was confirmed that the tumor size was significantly reduced in the group co-administered with L. Plantarum GB104 and GI-102 compared with the negative control group (Control). In addition, the group co-administered with L. Plantarum GB104 and GI-102 showed an excellent tumor size reduction effect compared with the group co-administered with L. Plantarum GB104 or GI-102 alone. Thus, it was confirmed that the anti-tumor efficacy was enhanced when L. Plantarum GB104 and GI-102 were co-administered.

[0306] Experimental Example 10 Inhibitory effect of triple administration of GB104 strain, immune anticancer agent GI-101 and chemical anticancer agent 5-FU on tumor growth

[0307] In this experimental example, the tumor therapeutic effect of triple administration of L. Plantarum GB104 strain, immune anticancer agent GI-101 and chemical anticancer agent (5-Fluorouracil, 5-FU) in the colorectal cancer MC-38 allograft model was confirmed. Six-week-old c57BL / 6 mice were received, and after a one-week adaptation period, the hair around the right flank was removed, and the experiment was performed at 7 weeks of age. The colorectal cancer cell line MC-38 derived from c57BL / 6 was transplanted at 2×10 5 The tumor model was established by subcutaneously injecting 100 μL of cells into the right flank of mice. The tumor size was measured using a digital calliper and calculated using the following formula: tumor volume (mm 3 )=(width 2 × length) / 2. On the 6th day after tumor cell injection, only tumors with a size of 10-30 mm were selected. 3 The mice were randomly divided into groups and then treated with L. Plantarum GB104 at a rate of 1 × 10 per mouse per day from day 6 until the end of the experiment. 9 The dose of CFU was orally administered to the animal model. The immune anticancer agent GI-101 was subcutaneously injected at 3 mg / kg once a week from day 6 for a total of 3 injections. The chemical anticancer agent 5-FU was intraperitoneally injected at 10 mg / kg once a week from day 6 for a total of 3 injections.

[0308] Fig.19 The graph shows the results of analyzing the synergistic effect by measuring tumor volume after triple administration of GB104, GI-101 and 5-FU in an allogeneic MC-38 mouse colorectal cancer model.

[0309] Fig. 20 The graph shows the comparative analysis results of measuring tumor volume after single and combined administration of GB104, GI-101 and 5-FU in an allogeneic MC-38 mouse colorectal cancer model.

[0310] like Fig.19 and Fig. 20 As shown, on the 26th day after tumor transplantation, compared with the negative control group (PBS), the GB104 single-administration group showed 29.72% tumor growth inhibition, and the GB104+GI-101 dual-administration group showed 50.01% tumor growth inhibition. In addition, an excellent anti-tumor effect of 63.92% was confirmed in the GB104+GI-101+5-FU triple-administration group.

[0311] This confirmed that in the treatment of colorectal cancer, the anti-tumor efficacy was enhanced by significantly inhibiting tumor growth not only in the group where the GB104 strain was administered alone, but also in the group where the immune anticancer agent GI-101 and the existing chemical anticancer agent 5-FU were administered in combination.

[0313] [Accession number]

[0314] Name of depository institution: Korea Institute of Biotechnology

[0315] Accession number: KCTC14107BP

[0316] Collection date: 20200114 Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure International Model The original of the deposit certificate issued by the following international depositary institution in accordance with Article 7.1 of the Treaty "To.GI Biome Company GI Biome Corporation 167, Songpa-daero Songpa-gu Seoul, South Korea"

Claims

1. A pharmaceutical composition for preventing or treating cancer, wherein: The composition comprises a first active substance and a second active substance as active ingredients, wherein the first active substance comprises a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and the second active substance comprises an immune anticancer agent as a first anticancer agent; or The composition comprises the first active substance as an effective ingredient, and the second active substance is administered in combination therewith.

2. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein: The first active substance has tumor growth inhibition or tumor metastasis inhibition activity.

3. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein: The first active substance induces apoptosis of cancer cells.

4. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein: The strain comprises the 16S rRNA of SEQ ID NO:

1.

5. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein: The strain is a strain deposited with the deposit number KCTC14107BP.

6. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein: The strain comprises a mutation of a naturally occurring Lactobacillus plantarum strain.

7. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein: The immune anticancer agent comprises a fusion protein of CD80 protein or a fragment thereof and IL-2 protein or a variant thereof.

8. The pharmaceutical composition for preventing or treating cancer according to claim 7, wherein: The IL-2 variant is substituted with at least one of amino acids 38, 42, 45, 61 and 72 of the amino acid sequence of SEQ ID NO:

4.

9. The pharmaceutical composition for preventing or treating cancer according to claim 7, wherein: The IL-2 variants have amino acid sequences of SEQ ID NO:5 to SEQ ID NO:

8.

10. The pharmaceutical composition for preventing or treating cancer according to claim 7, wherein: The CD80 fragment has the amino acid sequence of SEQ ID NO:

3.

11. The pharmaceutical composition for preventing or treating cancer according to claim 7, wherein: The fusion protein is composed of the following structural formula (I) or (II): N'-X-[Linker (1)]n-Fc domain-[Linker (2)]mY-C'(I) N'-Y-[Linker (1)]n-Fc domain-[Linker (2)]mX-C'(II) Wherein, in the structural formula (I) and (II), The N' is the N-terminus of the fusion protein, The C' is the C-terminus of the fusion protein, The X is CD80 protein or a fragment thereof, The Y is IL-2 protein or a variant thereof, The linker (1) and linker (2) are peptide linkers, The n and m are independently 0 or 1.

12. The pharmaceutical composition for preventing or treating cancer according to claim 7, wherein: The fusion protein has 90% or higher sequence identity with the amino acid sequence of SEQ ID NO: 12 to SEQ ID NO:

15.

13. The pharmaceutical composition for preventing or treating cancer according to claim 7, wherein: The fusion protein forms a homodimer through the combination of two identical proteins.

14. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein: The first active substance and the second active substance are administered simultaneously, sequentially or in reverse order.

15. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein: The first active substance and the second active substance are administered orally, intravenously, intraperitoneally or subcutaneously.

16. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein: The first active substance is administered orally, and the second active substance is administered intravenously or subcutaneously.

17. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein: The first active substance and the second active substance are administered to the individual at intervals of 1 to 10 days.

18. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein: The first active substance is administered to the individual at intervals of 1 to 3 days, and the second active substance is administered to the individual at intervals of 5 to 10 days.

19. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein: The second active substance further comprises a second anticancer agent, and the first anticancer agent and the second anticancer agent are different from each other.

20. The pharmaceutical composition for preventing or treating cancer according to claim 19, wherein: The second anticancer agent is any one selected from the following group: chemical anticancer agents, targeted anticancer agents and immune anticancer agents.

21. The pharmaceutical composition for preventing or treating cancer according to claim 20, wherein: The chemical anticancer agent is any one selected from the following group: an alkylating agent, a microtubule inhibitor, an antimetabolite, and a topoisomerase inhibitor.

22. The pharmaceutical composition for preventing or treating cancer according to claim 20, wherein: The chemical anticancer agent is any one selected from the following group: mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, thiotepa, altretamine, procarbazine, busulfan, streptozotocin, carmustine, lomustine, dacarbazine, cisplatin, carboplatin, oxaliplatin, docetaxel , Velban, Oncovin, Navelbine, 5-Fluorouracil, Capecitabine, Cytarabine, Gemcitabine, Fludarabine, Methotrexate, Pemetrexed, Mercaptopurine, Hycamtin, Camptosar, Vepesid, Paclitaxel, Blenoxane, Adriamycin, Etoposide and Cerubidine.

23. The pharmaceutical composition for preventing or treating cancer according to claim 20, wherein: The targeted anticancer agent targets any one protein selected from the following group: epidermal growth factor receptor, vascular growth factor receptor (VEGFR), CD20, CD38, RNAK-L, BTK, Bcr-abl, PDGFR / FGFR series, MEK / RAF, HER2 / Neu, ubiquitin, JAK, MAP2K, ALK, PARP, tumor growth factor β receptor, proteasome, Bcl-2, C-Met, VR1, VR2, VR3, c-kit, AXL, RET, Braf, DNA methyltransferase, CDK4 / 6 and STING.

24. The pharmaceutical composition for preventing or treating cancer according to claim 20, wherein: The targeted anticancer agent is any one selected from the following group: Olaratumab, Erlotinib, Panitumumab, Trastuzumab, Trastuzumab-emtansine, Pertuzumab, Cetuximab, Rituximab, Bevacizumab, Axitinib, Lenvatinib, Ramucirumab, Aflibercept, Aflibercept, Obinutuzumab, Daratumumab, Denosumab, Ibrutinib, Dasatinib, Radotinib, Nilotinib, Imatinib, Bosutinib, Galunisertib, Vactosertib, Nintedanib, Sunitinib, Sorafenib ), cabozantinib, Regorafenib, Masitinib, Semaxanib, Ceritinib, Tivozanib, Brigatinib, Vandetanib, Pazopanib, Trametinib, Temsirolimus, Dabrafenib, Dacomitinib, Afatinib, Lapatinib inib), Neratinib, Lenalidomide, Osimertinib, Ixazomib, Olmutinib, Everolimus, Ruxolitinib, Lestaurtinib, Pacritinib, Cobimetinib, Selumetinib, Binimetinib, Bortezomib, Alectinib,Crizotinib, Venetoclax, Bemcentinib, Gilteritinib, Selpercatinib, Pralsetinib, Vemurafenib, Olaparib, Talazoparib, Niraparib, Rucaparib, Azacitidine, Decitabine, Guadecitabine, Gefitinib, Abemaciclib, Ribociclib, Palbociclib and DMXAA.

25. The pharmaceutical composition for preventing or treating cancer according to claim 20, wherein: The immuno-anticancer agent is directed against any one antibody selected from the following group: 2B4, 4-1BB (CD137), AaR, B7-H3, B7-H4, BAFFR, BTLA, CD2, CD7, CD27, CD28, CD30, CD40, CD80, CD83 ligand, CD86, CD160, CD200, CDS, CEACAM, CTLA-4, GITR, HVEM, ICAM-1, KIR, LAG-3, LAIR1, LFA-1 (CD11a / CD18), LIGHT, NKG2C, NKp80, OX40, PD-1, PD-L1, PD-L2, SLAMF7, TGFRp, TIGIT, Tim3 and VISTA.

26. The pharmaceutical composition for preventing or treating cancer according to claim 20, wherein: The immunotherapy anticancer agent is any one selected from the following group: Atezolizumab, Avelumab, Durvalumab, Nivolumab, Pembrolizumab, Abagovomab, Adecatumumab, Afutuzumab, Alemtuzumab, Anatumomab mafenatox), mepolizumab, blinatumomab, BMS-936559, catumaxomab, cemiplimab, epacadostat, epratuzumab, indoximod, inotuzumab, okitamostat, inotuzumab Ozogamicin, intelumumab, ipilimumab, isatuximab, lambalizumab, MED14736, MPDL3280A, obinutuzumab, ocaraituzumab, ofatumumab, olatatumab, pidilizumab, rituximab, ticilizumab, samalizumab, and tremelimumab.

27. The pharmaceutical composition for preventing or treating cancer according to claim 19, wherein: The first anticancer agent and the second anticancer agent are administered simultaneously, sequentially or in reverse order.

28. The pharmaceutical composition for preventing or treating cancer according to claim 19, wherein: The first anticancer agent and the second anticancer agent are administered orally, intravenously, intraperitoneally or subcutaneously.

29. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein: The cancer is any one selected from the following group: gastric cancer, liver cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, gallbladder cancer, bile duct cancer, cervical cancer, thyroid cancer, laryngeal cancer, acute myeloid leukemia, brain tumor, neuroblastoma, retinoblastoma, salivary gland cancer, melanoma, bladder cancer, kidney cancer, blood cancer, esophageal cancer, head and neck cancer, skin cancer, small intestine cancer, anal cancer, colon cancer, rectal cancer and lymphoma.

30. The pharmaceutical composition for preventing or treating cancer according to claim 20, wherein: The colorectal cancer occurs at any one site selected from the following group: ascending colon, transverse colon, descending colon, sigmoid colon and rectal mucosa.

31. A health functional food for preventing or improving cancer, wherein: The health functional food comprises a first active substance and a second active substance as effective ingredients, wherein the first active substance comprises a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and the second active substance comprises an immune anticancer agent as a first anticancer agent; or The health functional food contains the first active substance as an effective ingredient, and the second active substance is administered in combination with the first active substance.

32. The health functional food for preventing or improving cancer according to claim 31, wherein: The strain is a strain deposited with the deposit number KCTC14107BP.

33. The health functional food for preventing or improving cancer according to claim 31, wherein: The immune anticancer agent comprises a fusion protein of IL-2 protein or its variant and CD80 protein or its fragment.

34. The health functional food for preventing or improving cancer according to claim 31, wherein: The second active substance further comprises a second anticancer agent, and the first anticancer agent and the second anticancer agent are different from each other.

35. The health functional food for preventing or improving cancer according to claim 34, wherein: The second anticancer agent is any one selected from the following group: chemical anticancer agents, targeted anticancer agents and immune anticancer agents.

36. The health functional food for preventing or improving cancer according to claim 31, wherein: The first active substance and the second active substance are administered simultaneously, sequentially or in reverse order.

37. The health functional food for preventing or improving cancer according to claim 31, wherein: The first active substance and the second active substance are administered orally, intravenously, intraperitoneally or subcutaneously.

38. The health functional food for preventing or improving cancer according to claim 31, wherein: The first active substance is administered orally, and the second active substance is administered intravenously or subcutaneously.

39. The health functional food for preventing or improving cancer according to claim 31, wherein: The cancer is any one selected from the following group: gastric cancer, liver cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, gallbladder cancer, bile duct cancer, cervical cancer, thyroid cancer, laryngeal cancer, acute myeloid leukemia, brain tumor, neuroblastoma, retinoblastoma, salivary gland cancer, melanoma, bladder cancer, kidney cancer, blood cancer, esophageal cancer, head and neck cancer, skin cancer, small intestine cancer, anal cancer, colon cancer, rectal cancer and lymphoma.

40. The pharmaceutical composition for preventing or treating cancer according to claim 39, wherein The colorectal cancer occurs at any one site selected from the following group: ascending colon, transverse colon, descending colon, sigmoid colon and rectal mucosa.

41. A method for preventing or treating cancer, the method comprising the following steps: A composition is administered to an individual in need thereof, wherein the composition comprises a first active substance and a second active substance as active ingredients, wherein the first active substance comprises an effective amount of a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and the second active substance comprises a first anticancer agent as an immune anticancer agent; or, the composition comprises the first active substance as an active ingredient, and the second active substance is administered in combination with the first active substance.

42. Use of a composition in the preparation of a pharmaceutical preparation for preventing or treating cancer, wherein: The composition comprises a first active substance and a second active substance as effective ingredients, wherein the first active substance comprises a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and the second active substance comprises a first anticancer agent as an immune anticancer agent; or, the composition comprises the first active substance as an effective ingredient, and the second active substance is administered in combination with the first active substance.

43. Use of a composition in preparing a health functional food for preventing and treating cancer, wherein: The composition comprises a first active substance and a second active substance as effective ingredients, wherein the first active substance comprises a Lactobacillus plantarum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and the second active substance comprises a first anticancer agent as an immune anticancer agent; or, the composition comprises the first active substance as an effective ingredient, and the second active substance is administered in combination with the first active substance.

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