Combination therapy with Lactobacillus plantarum strains and cancer treatment method using the same

Abstract

This invention relates to a pharmaceutical composition for cancer treatment in which a Lactobacillus plantarum strain is administered in combination with an anticancer agent. The Lactobacillus plantarum strain or a culture medium of the strain can not only inhibit the proliferation of cancer cells or induce their death, but also exhibits a synergistic effect in inhibiting the proliferation of cancer cells and tumors when administered in combination with an anticancer agent, and can be usefully used in the form of a pharmaceutical composition or a health functional food for the prevention, treatment, or amelioration of cancer.

Description

[Technical Field]

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

[0002] The term "microbiome" is a combination of the words "microbe" and "genome," and the term "human microbiome" refers to the genomes of all microorganisms living in the human body. While various types of microorganisms are present in several parts of the human body, 70% of the human microbiome is distributed in the digestive tract, with the largest number of microorganisms residing in the large intestine. The microbiome is closely related to the human immune system and physical development, and its strong association with various diseases, including metabolic disorders such as obesity and diabetes, inflammatory bowel disease, and depression, has been steadily reported. After the initial completion of the Human Genome Project in 2002, attempts to identify the relationship between genes and intractable diseases were made, and it was found that environmental factors, particularly the microbiome environment, may have a significant impact on disease. Microbiome therapeutics grow in the intestines as living organisms and affect the human body through interactions with human cells. For this reason, they are attracting significant attention as innovative new drugs that can improve the low efficacy and high recurrence rates of existing drugs for intractable diseases. In addition, in December 2022, the US FDA approved Ferring Pharmaceuticals' REBYOTA (registered trademark), the first microbiome-based treatment for C. difficile infection (Clostridium difficile infection, CDI), marking the beginning of the practical application of microbiome therapeutics. Seres Therapeutics' SER-109 is also currently under FDA review as the first oral microbiome therapeutic.

[0003] Cancer is considered one of the major health issues worldwide. It is the second leading cause of death worldwide and, once it develops, requires intensive treatment and thorough management, placing a significant burden on individuals and socioeconomically. As of 2021, the global anti-cancer drug market was worth approximately KRW 225 trillion, and is expected to grow at an average annual rate of 11% to reach KRW 470 trillion by 2028 due to factors such as an increase in the number of patients, growth in immune-based anti-cancer drugs, and the emergence of new treatments.

[0004] Traditional cancer treatment involves a combination of chemotherapy, surgery, hormonal therapy, and / or radiation therapy to eliminate a patient's neoplastic cells. Cancer chemotherapy is preferably based on the use of drugs that kill replicating cells faster than the drug kills the patient's normal cells. Anticancer drugs exert cytotoxicity by interfering with the metabolic pathways of cancer cells and directly interacting with DNA, blocking DNA replication, transcription, and translation processes or interfering with the synthesis of nucleic acid precursors, thereby inhibiting cell division. Therefore, these anticancer drugs not only selectively act on cancer cells but also fatally damage normal cells, especially actively dividing tissue cells, causing various serious side effects such as vomiting, bone marrow depression, gastrointestinal disorders, alopecia, diarrhea, and liver and kidney toxicity. In response to this, immune anticancer drugs (or immune checkpoint inhibitors) have attracted much attention as third-generation anticancer drugs, but their usability is very limited. Treatment efficacy is generally less than 30%. Therefore, to fully conquer cancer, further research into the mechanisms by which gut microbes enhance the efficacy of immune anticancer drugs is needed.

[0005] Recently, combination therapies have become popular to enhance the efficacy of disease treatments. Therefore, there is a need to develop combination therapies that can synergistically increase the effects of single-component treatments. Summary of the Invention [Problem to be solved by the invention]

[0006] One aspect is to provide a pharmaceutical composition for preventing or treating cancer, which comprises as active ingredients a first active substance comprising a Lactobacillus plantarum strain, a culture of the strain, a disrupted product of the strain, or a mixture thereof, and a second active substance comprising an immunological anticancer agent that is a first anticancer agent, or which comprises the first active substance as an active ingredient and is administered in combination with the second active substance.

[0007] Another aspect is to provide a health functional food for preventing or ameliorating cancer, which contains as active ingredients a first active substance comprising a Lactobacillus plantarum strain, a culture of said strain, or a disrupted product of said strain, or a mixture thereof, and a second active substance comprising an immunological anticancer agent that is a first anticancer agent, or which contains the first active substance as an active ingredient and is administered in combination with the second active substance.

[0008] Another aspect is to provide a kit for preventing or treating cancer, which comprises as active ingredients a first active substance comprising a Lactobacillus plantarum strain, a culture of said strain, a homogenate of said strain, or a mixture thereof, and a second active substance comprising a first anti-cancer agent, which is an immunological anti-cancer agent, or alternatively, the first active substance is comprised as an active ingredient and the second active substance is administered in combination.

[0009] Another aspect is to provide a method for delivering a drug into an individual, comprising the step of administering to an individual in need thereof a composition containing as active ingredients a first active substance comprising a Lactobacillus plantarum strain, a culture of said strain, a homogenate of said strain, or a mixture thereof, and a second active substance comprising a first anti-cancer agent, which is an immunosuppressant anti-cancer agent, or alternatively, the first active substance as an active ingredient and the second active substance in combination.

[0010] Another embodiment is a method for treating a stomach ulcer by administering an effective amount of Lactobacillus plantarum. The present invention provides a method for preventing or treating cancer, comprising the step of administering to an individual in need thereof a composition containing, as active ingredients, a first active substance comprising a Bacillus subtilis (Bacillus plantarum) strain, a culture of the strain, a disruptant of the strain, or a mixture thereof, and a second active substance comprising a first anti-cancer agent, which is an immunosuppressive anti-cancer agent, or a composition containing, as an active ingredient, the first active substance and the second active substance administered in combination.

[0011] Another aspect provides use of a composition comprising as active ingredients a first active substance comprising a Lactobacillus plantarum strain, a culture of the strain, a homogenate of the strain, or a mixture thereof, and a second active substance comprising an immunological anti-cancer agent that is the first anti-cancer agent, or a composition comprising the first active substance as an active ingredient and the second active substance administered in combination, for the production of a pharmaceutical preparation for preventing or treating cancer or a functional health food. [Means for solving the problem]

[0012] One aspect provides a Lactobacillus strain, specifically the Lactobacillus plantarum GB104 strain, having anti-cancer activity, for example, anti-cancer activity against colon cancer.

[0013] Lactobacillus is a type of aerobic or Lactobacillus is a facultative anaerobic Gram-positive bacillus. Microorganisms belonging to the Lactobacillus genus include Lactobacillus plantarum and Lactobacillus sakei. The present inventors conducted research to develop a new strain with excellent anti-cancer effects and selected Lactobacillus plantarum GB104 as a candidate anti-cancer strain. The strain was deposited at the Korea Institute of Bioscience and Biotechnology's Biological Resources Center on January 14, 2020, under the accession number KCTC14107BP. The strain corresponds to a probiotic strain and is harmless to the human body, so it can be used without side effects.

[0014] The Lactobacillus species has been renamed Limosilactobacillus or Lactiplantibacillus, and the modified strain names can be used interchangeably herein. For example, Lactobacillus plantarum has been renamed Lactiplantibacillus plantarum.

[0015] As used herein, the term "Lactobacillus plantarum GB104" may be used in conjunction with the L. Plantarum GB104 strain or the Lactobacillus plantarum GB104 strain (accession number: KCTC14107BP).

[0016] In one embodiment, the strain may be the strain deposited under accession number KCTC14107BP.

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

[0018] In one embodiment, the strain may have a 16S rRNA comprising the nucleotide sequence of SEQ ID NO: 1 or a nucleotide sequence having 97% or more identity thereto, specifically at least 93%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.8%, 99.9%, or 100% identity to the nucleotide sequence of SEQ ID NO: 1 herein.

[0019] In one embodiment, the strain may be a mutant strain of a naturally occurring strain.

[0020] In one embodiment, the bacterial strain may be a live bacterium, a killed bacterium, or a cytoplasmic fraction obtained by disrupting the bacterial strain, preferably a live bacterium.

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

[0022] As used herein, the term "lysate" can be used interchangeably with "lysate" to refer to a solution or suspension of broken down microbial cells, such as Lactobacillus plantarum, in an aqueous medium. Cell lysates contain, for example, DNA, RNA, and proteins. The lysate contains macromolecules such as proteins, peptides, carbohydrates, lipids, etc. and / or small molecules such as amino acids, sugars, fatty acids, etc., or fractions thereof. Furthermore, the lysate contains cell debris which may be smooth or granular in structure.

[0023] The culture medium may include the culture medium itself obtained by culturing the strain, a concentrate thereof, or a freeze-dried product thereof, or a culture supernatant obtained by removing the strain from the culture medium, or a concentrate or freeze-dried product thereof.

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

[0025] In one embodiment, the first active substance comprising one or more selected from the group consisting of the strain, a culture of the strain, a disrupted product of the strain, or an extract of the strain, culture, and disrupted product may have anti-cancer activity.

[0026] Specifically, the anti-cancer activity may be the activity of delaying the onset of tumors or inhibiting the rate of tumor growth.

[0027] In one embodiment, the first active agent may have tumor growth inhibitory or tumor metastasis inhibitory activity.

[0028] In one embodiment, the first active agent may induce cell death in cancer cells.

[0029] According to one embodiment, the anti-cancer activity may be a cancer cell growth inhibitory activity in which the survival rate of cancer cells is reduced by 90% to 98% when colon cancer cells HCT116 are treated with the supernatant of Lactobacillus plantarum GB104 strain, or an activity in which the tumor volume is reduced by 10% to 90% when the Lactobacillus plantarum GB104 strain is administered to mice subcutaneously implanted with MC-38 or HT-29 cancer cell lines, compared to a control group not administered the strain.

[0030] According to one embodiment, the anticancer activity is an activity that induces cell death in cancer cells, and when colon cancer cells HCT116 are treated with the supernatant of Lactobacillus plantarum GB104 strain, the induction of early cell death in cancer cells is increased 5 to 9 times compared to a control group that is not administered the strain.

[0031] In one embodiment, the first active agent may have anti-cancer activity, particularly anti-cancer activity against colon cancer.

[0032] The term "cancer," as used herein, refers to a physiological condition in an animal that is typically characterized by abnormal or uncontrolled cell growth. Cancer and cancer pathology can be associated with, for example, metastasis, interference with normally functioning surrounding cells, release of abnormal levels of cytokines or other secretory products, suppression or augmentation of inflammatory or immunological responses, neoplasia, premalignancy, malignancy, invasion of surrounding or distant tissues or organs, e.g., lymph node invasion, and the like.

[0033] The cancer can be a gastrointestinal cancer or a non-gastrointestinal cancer.

[0034] The gastrointestinal cancer is a malignant tumor that occurs 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 group consisting of esophageal cancer, gallbladder cancer, liver cancer, biliary tract cancer, pancreatic cancer, stomach cancer, small intestine cancer, colon cancer, anal cancer, and rectal cancer, but is not limited to these, and one example may be colon cancer.

[0035] The non-gastrointestinal cancer includes, but is not limited to, malignant tumors occurring in organs other than the gastrointestinal tract or 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 one embodiment, the cancer may be any selected from the group consisting of gastric cancer, liver cancer, lung cancer, colon cancer, breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, gallbladder cancer, biliary tract cancer, cervical cancer, thyroid cancer, laryngeal cancer, acute myeloid leukemia, brain cancer, 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 those arising in one or more sites selected from the group consisting of the ascending colon, transverse colon, descending colon, sigmoid colon, and rectal mucosa, and may be one or more types selected from the group consisting of, but not limited to, adenocarcinoma, lymphoma, malignant carcinoid tumor, leiomyosarcoma, Kaposi's sarcoma, and squamous cell carcinoma.

[0038] In one embodiment, the first active substance may comprise a Lactobacillus plantarum strain as the sole active ingredient, or may comprise one or more pharmaceutically acceptable carriers, excipients or diluents.

[0039] In one embodiment, the second active agent can include an anti-cancer agent. More specifically, the second active agent can include two different anti-cancer agents, a first anti-cancer agent and a second anti-cancer agent.

[0040] The anticancer agent may be used in combination with conventional therapies selected from the group consisting of chemotherapeutic agents, targeted anticancer agents, anticancer viruses, antibody therapeutic agents, immunological anticancer agents (hereinafter, the term "immune checkpoint inhibitors" may be used interchangeably), and combinations thereof. Specifically, the anticancer agent may comprise an antibody, an antigen-binding fragment, or a fusion protein comprising 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 Fc region, and the one or more single-domain antibodies, or antigen-binding fragments thereof, may be non-covalently or covalently conjugated to an antigen.

[0041] In one embodiment, the second active agent may comprise an immunological anti-cancer agent.

[0042] The term "antibody" as used herein refers to any isotype of intact immunoglobulin or antigen-binding fragment that can compete with the intact antibody for binding to a target antigen. Examples include chimeric, humanized, fully human, and bispecific antibodies, or antigen-binding fragments thereof. The antibody itself is a type of antigen-binding protein. The antibody or antigen-binding fragment thereof can be derived from a single source or from a chimera. The chimeric antibody contains portions derived from two different antibodies and is described in more detail below. The antibody or antigen-binding fragment thereof can be derived from a hybridoma, recombinant DNA technology, or enzymatic or recombinant DNA technology of an intact antibody. can be produced by chemical cleavage. Unless otherwise specified, the term "antibody" herein can include antibodies comprising two full-length heavy chains and two full-length light chains, as well as derivatives, variants, fragments and mutants thereof.

[0043] As used herein, the term "light chain" includes a full-length light chain having sufficient variable region sequence to provide binding specificity to an antigen or epitope and fragments thereof. The full-length light chain includes a variable region domain, VL, and a constant region domain, CL. The light chain variable region domain is located at the amino terminus of the light chain polypeptide. Types of light chains include kappa and lambda chains.

[0044] As used herein, the term "heavy chain" includes a full-length heavy chain having sufficient variable region sequence to provide binding specificity to an antigen or epitope and fragments thereof. The full-length heavy chain includes a variable region domain, VH, and three constant region domains, CH1, CH2, and CH3. The VH domain is located at the amino terminus of the heavy chain polypeptide, the CH domain is located at the carboxyl terminus, and CH3 is closest to the carboxyl terminus. Heavy chains include IgG (including IgG1, IgG2, IgG3, and IgG4 subtypes), IgA (including IgA1 and IgA2 subtypes), and IgM and IgE isotypes.

[0045] As used herein, an "antigen-binding fragment" of an antibody or immunoglobulin chain (heavy or light chain) includes a portion of the antibody that lacks some amino acids compared to the full-length chain but is capable of specifically binding to an antigen. This fragment can be considered biologically active in terms of its ability to specifically bind to a target antigen or compete with other antibodies or antigen-binding fragments for binding to a particular epitope. In one aspect, the fragment contains at least one CDR present in a full-length light or heavy chain, and in some embodiments, a shortened heavy and / or light chain or portion thereof. This biologically active fragment can be produced by recombinant genetic techniques, for example, by enzymatic or chemical cleavage of an intact 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 may be derived from any mammal, including, but not limited to, human, mouse, rat, camelid, or rabbit.

[0046] As used herein, the "Fc" region comprises two heavy chain fragments comprising the CH2 and CH3 domains of an antibody, which are held together by two or more disulfide bonds and hydrophobic interactions of the CH3 domain.

[0047] As used herein, a "Fab fragment" consists of one light chain and one heavy chain containing only the variable region and CH1. The heavy chain of a Fab molecule cannot form disulfide bonds with other heavy chain molecules. An scFab is two Fab molecules linked by a flexible linker.

[0048] As used herein, a "Fab' fragment" includes a Fab fragment plus the region between the CH1 and CH2 domains of the heavy chain, and is capable of forming disulfide bonds between the two heavy chains of two molecules of the Fab' fragment to generate an F(ab')2 molecule.

[0049] The term "F(ab')2" as used herein refers to two light chains and the variable region, CH1, and the constant region between the CH1 and CH2 domains as mentioned above. The F(ab')2 fragment contains two heavy chains containing portions of the Fab' fragment, which results in the formation of disulfide bonds between the two chains. Thus, the F(ab')2 fragment is composed of two Fab' fragments, which are held together by disulfide bonds between them.

[0050] As used herein, the term "Fv region" refers to an antibody fragment that contains the variable regions of the heavy and light chains but does not contain the constant region. An sdFv is a fragment in which the heavy and light chains are linked by a disulfide bond. An scFc is a fragment in which the single-stranded variable regions (Fv) of the heavy and light chains are linked via a flexible linker. An scFv-Fc is a fragment in which the Fc is linked to the scFv. A minibody is a fragment in which the CH3 is linked to the scFv. A diabody contains the scFvs of both molecules.

[0051] As used herein, a "short-chain antibody (scAb)" is a single polypeptide chain comprising one variable region of a heavy or light chain constant region, with the heavy and light chain variable regions connected by a flexible linker.

[0052] A "domain antibody (dAb)" herein is an immunologically functional immunoglobulin fragment containing only the variable region of a heavy chain or the variable region of a light chain.

[0053] As used herein, a "bivalent antigen-binding protein" or "bivalent antibody" comprises two antigen-binding sites. The two antigen-binding sites in such a bivalent antibody may have the same antigen specificity, or they may be bispecific antibodies that each bind to a different antigen.

[0054] As used herein, a "multi-specific antigen-binding protein" or "multi-specific antibody" is one that targets more than one antigen or epitope.

[0055] As used herein, a "bispecific" or "dual-specific" antigen-binding protein or antibody is a hybrid antigen-binding protein or antibody having two different antigen-binding sites. Such bispecific antibodies are a type of multispecific antigen-binding protein or multispecific antibody and can be produced by various known methods, such as hybridoma fusion or linkage of Fab' fragments.

[0056] In one embodiment, the additional polypeptide comprises an additional antibody or fragment thereof, such as, for example, intact IgG, IgE and IgM, bispecific or multispecific antibodies (such as, for example, Zybodies®), single chain Fv, polypeptide-Fc fusions, Fab, cameloid antibodies, masked antibodies (such as, for example, Probodies®), Small Modular ImmunoPharmaceuticals ("SMIPs™"), single chain or tandem diabodies (TandAb®), VHHs (including, but not limited to, those described in this disclosure), Anticalin®, Nanobodies®, minibodies, BiTE®, ankyrin repeat proteins or DARPIN®, Avimers®, DART, TCR mimetic antibodies, Adnectins®, Affilins®, Trans-bodies®, Affibodies®, TrimerX. (R), MicroProteins, Fynomers®, Centyrins®, and KALBITOR®. Additional antibodies also target PD-1, TIM-3, LAG-3, IDO, A2AR, TGF-β, CD47, or another protein involved in an immunosuppressive pathway.

[0057] In one embodiment, the additional polypeptide comprises or consists of part or all of a tumor-associated antigen (TAA) or tumor-specific antigen (TSA). Non-limiting examples of TSA or TAA antigens include differentiation antigens such as MART-1 / MelanA (MART-I), gp100 (Pmel17), tyrosinase, TRP-1, TRP-2, and tumor-specific multilineage antigens such as MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, and p15; overexpressed embryonic antigens such as CEA; overexpressed oncogenes and mutated tumor suppressor genes such as p53, Ras, and HER-2 / neu; unique tumor antigens derived from chromosomal translocations such as BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, and 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, CA19-9, CA72-4, CAM17.1, NuMa, K-ras, β-catenin, CDK4, Mum-1, p15, p16, 43-9F, 5T4, 791Tgp72, α-fetoprotein, β-HCG, BCA225, BTAA, CA125, CA15-3\CA27.29\BCAA, CA195, CA242, CA-50, CAM43, CD68\P1, CO-029, FGF-5, G250, Ga733\E These include pCAM, 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, TAAL6, TAG72, TLP, MUC16, IL13Rα2, FRα, VEGFR2, Lewis Y, FAP, EphA2, CEACAM5, EGFR, CA6, CA9, GPNMB, EGP1, FOLR1, endothelial receptors, STEAP1, SLC44A4, nectin-4, AGS-16, guanylate cyclase-C, MUC-1, CFC1B, integrin α3 chain (a3b1, laminin receptor chain), and TPS.

[0058] In one embodiment, the additional polypeptide comprises or consists of 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, glypican 3 (GPC3), mesothelin CD33 / IL3Ra, c-Met, PSCA, PSMA, glycolipid F77, EGFRvIII, GD-2, MY-ESO-1, and MAGEA3.

[0059] In one embodiment, the immunological anti-cancer agent can comprise a fusion protein comprising a membrane protein involved in immune regulation and a cytokine. For example, the fusion protein can comprise a membrane protein or a fragment thereof, an Fc region, and a cytokine or a variant thereof.

[0060] The term "membrane protein" as used herein refers to a protein inserted into or attached to the surface of a cell membrane consisting of a lipid bilayer, which binds to a ligand to transmit costimulatory and coinhibitory responses and is involved in immune regulation. In one embodiment, 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 a B Also known as CD80, CD86 is a transmembrane protein expressed on the surface of T cells, B cells, dendritic cells, and monocytes. CD80 is known to bind to CD28, CTLA4 (CD152), and PD-L1. CD80, CD86, CTLA4, and CD28 participate in the costimulatory-coinhibitory system, which regulates, for example, T cell activity and is involved in proliferation, differentiation, and survival.

[0061] The term "cytokine" as used herein refers to small proteins of approximately 5 to 20 kDa that are important in cell signaling and are produced by various cells, including immune cells. These proteins act via cellular receptors to regulate the maturation, differentiation, and activity of specific immune cell populations, thereby regulating the balance of immune responses. In one embodiment, the cytokine may be an interleukin, more specifically, IL-2 or a variant thereof. IL-2 stimulates the proliferation and differentiation of T cells and induces the production of cytotoxic T lymphocytes (CTLs) and the differentiation of peripheral blood lymphocytes into cytotoxic cells and lymphokine-activated killer cells (LAK cells). IL-2 is involved in the proliferation and differentiation of B cells, promotes immunoglobulin synthesis by B cells, and stimulates the production, proliferation, and activation of natural killer cells (NK cells).

[0062] In one embodiment, the immunological anti-cancer agent can be a fusion protein comprising CD80 and IL-2 proteins. More specifically, the immunological anti-cancer agent can be a fusion protein comprising CD80 protein or a fragment thereof and IL-2 protein or a variant thereof.

[0063] 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 one embodiment, the CD80 fragment may be a CD80 signal sequence, excluding amino acids 1 to 34 from the N-terminus. Specifically, the CD80 fragment may be a protein consisting of amino acids 35 to 242 of SEQ ID NO: 2. In one embodiment, the CD80 fragment may have the amino acid sequence of SEQ ID NO: 3.

[0064] The IL-2 may be wild-type IL-2 or a mutant thereof. The wild-type IL-2 may have the amino acid sequence of SEQ ID NO: 44. The IL-2 mutant may have some of the amino acids of wild-type IL-2 substituted. In one embodiment, the IL-2 mutant formed by amino acid substitution may have at least one of the amino acids at positions 38, 42, 45, 61, and 72 substituted in the amino acid sequence of SEQ ID NO: 4.

[0065] Specifically, the IL-2 mutant 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.

[0066] Preferably, one embodiment of the IL-2 mutant may be one in which any combination of substitutions selected from the following combinations a to d has occurred in the amino acid sequence of SEQ ID NO: 4: (a) R38A / F42A (b) R38A / F42A / Y45A (c) R38A / F42A / E61R (d) R38A / F42A / L72G

[0067] In one embodiment, the IL-2 mutant has the amino acid sequence of SEQ ID NO: 5 to 8. It can be something.

[0068] The CD80 or fragment thereof and IL-2 or a variant thereof may be linked by a linker or a carrier. The terms "linker" and "carrier" may be used interchangeably herein. The linker connects the two proteins. An example of the linker may include 1 to 50 amino acids, albumin or a fragment thereof, or an immunoglobulin Fc domain. The immunoglobulin Fc domain refers to a protein that includes immunoglobulin heavy chain constant region 2 (CH2) and heavy chain constant region 3 (CH3), but does not include immunoglobulin heavy and light chain variable regions and light chain constant region 1 (CH1). The immunoglobulin may be IgG, IgA, IgE, IgD, or IgM, preferably IgG4. The wild-type immunoglobulin G4 Fc domain may have the amino acid sequence of SEQ ID NO: 9.

[0069] Specifically, the fusion protein may have a structure in which CD80 and IL-2 proteins, or IL-2 and CD80, are linked to the N-terminus and C-terminus, respectively, of the Fc domain as a linker (or carrier). The link between the N-terminus or C-terminus of the Fc domain and CD-80 or IL-2 may be optionally achieved via a linker peptide.

[0070] In one embodiment, the fusion protein may be 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) In this case, in the structural formulas (I) and (II), the N' is the N-terminus of the fusion protein; the C' is the C-terminus of the fusion protein; X is a CD80 protein or a fragment thereof; Y is an IL-2 protein or a variant thereof; said linker 1 and linker 2 are peptide linkers; The n and m each independently represent 0 or 1.

[0071] Preferably, the fusion protein may be of structural formula (I): The CD80 protein or a fragment thereof and the IL-2 protein or a variant thereof are the same as those described above.

[0072] A peptide linker 1 can be included between the CD80 protein and the Fc domain. The peptide linker 1 can consist of 5 to 80 consecutive amino acids, 20 to 60 consecutive amino acids, 25 to 50 consecutive amino acids, or 30 to 40 amino acids. In one embodiment, the peptide linker 1 can consist of 30 amino acids. Furthermore, the peptide linker 1 can contain at least one cysteine. Specifically, it can contain one, two, or three cysteines. Furthermore, the peptide linker 1 can be derived from an immunoglobulin hinge. In one embodiment, the peptide linker 1 can be a peptide linker consisting of the amino acid sequence of SEQ ID NO: 10.

[0073] The peptide linker 2 can consist of 1 to 50 consecutive amino acids, or 3 to 30 consecutive amino acids, or 5 to 15 amino acids. In one embodiment, the peptide linker 2 can be (G4S)n (where n is an integer from 1 to 10). In this case, n in (G4S)n can be 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In one embodiment, the peptide linker 2 can be a peptide linker consisting of the amino acid sequence of SEQ ID NO: 11.

[0074] In one embodiment, the fusion protein may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NOs: 12 to 15. More specifically, the fusion protein may have the amino acid sequence of SEQ ID NOs: 12 to 15.

[0075] In one embodiment, the fusion protein may be encoded by a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 17 to 20. More specifically, the fusion protein may have the nucleic acid sequence of SEQ ID NO: 17 to 20.

[0076] The polynucleotide encoding the fusion protein may contain a signal sequence The signal sequence may further comprise a nucleic acid encoding a signal sequence or leader sequence. As used herein, the term "signal sequence" refers to a signal peptide that directs the secretion of a protein of interest. The signal peptide is cleaved after translation in a host cell. Specifically, the signal sequence is an amino acid sequence that initiates the translocation of a protein through the endoplasmic reticulum (ER) membrane. In one embodiment, the signal sequence may have the amino acid sequence of SEQ ID NO: 16.

[0077] In this case, the dimer can be formed by a disulfide bond between the fusion proteins constituting the dimer, which is formed by cysteines present in the linker. More specifically, the fusion proteins constituting the dimer can be proteins having the amino acid sequences of SEQ ID NOs: 12 to 14.

[0078] In one embodiment, the second active substance may further comprise a second anticancer drug, wherein the first anticancer drug and the second anticancer drug contained in the second active substance may be different from each other.

[0079] In one embodiment, the second anti-cancer agent may be any one selected from the group consisting of a chemical anti-cancer agent, a targeted anti-cancer agent, and an immune anti-cancer agent.

[0080] The term "chemo-anticancer agent" as used herein is also referred to as an antitumor agent or a cytotoxic agent. It is a general term for drugs that exhibit anticancer activity by acting directly on DNA to block the replication, transcription, and translation processes of DNA or by interfering with the synthesis of nucleic acid precursors in metabolic pathways, thereby inhibiting cell division. Antitumor drugs exhibit cytotoxicity by acting not only on tumor cells but also on normal cells. Chemo-anticancer agents can be used in maintenance therapy. Furthermore, the term "maintenance therapy" as used herein refers to a treatment method using drugs to treat cancer after initial anticancer therapy, and is performed to prevent or delay cancer recurrence.

[0081] Specifically, the anticancer chemotherapy agent may be any one selected from the group consisting of alkylating agents, microtubule inhibitors, antimetabolites, and topoisomerase inhibitors. The alkylating agents include mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, thiotepa (T). The anti-metabolite may be any one selected from the group consisting of hiotepa, altretamine, procarbazine, busulfan, streptozocin, carmustine, lomustine, dacarbazine, cyclophosphamide (cisplatin), carboplatin, and oxaliplatin. The microtubule inhibitor may be any one selected from the group consisting of docetaxel, velban, oncovin, and navelbine. The antimetabolite may be any one selected from the group consisting of 5-fluorouracil (Fluorouracil). The topoisomerase inhibitor may be any one selected from the group consisting of cyclosporine, capecitabine, cytarabine, gemcitabine, fludarabine, methotrexate, pemetrexed, and mercaptopurine. The topoisomerase inhibitor may be any one selected from the group consisting of hycamtin, camptosar, vepesid, paclitaxel, blenoxane, adriamycin, etoposide, and cerubidine.

[0082] The term "targeted anticancer drug" as used herein refers to a therapeutic agent that specifically kills cancer cells by targeting specific proteins or specific genetic alterations that are abundant only in cancer cells and blocking signals involved in the growth and development of cancer. Targeted anticancer drugs are classified into monoclonal antibodies that react extracellularly and small molecule substances that act intracellularly. Monoclonal antibodies are anticancer drugs that block cancer cell-inducing signals transmitted extracellularly and act on initiating signals related to proliferation, death, etc., while small molecule substances act on complex signal transduction that occurs intracellularly.

[0083] Specifically, target proteins may include epidermal growth factor receptor (EGFR), vascular endothelial growth factor receptor (VEGFR), CD20, CD38, RNAK-L, BTK, Bcr-abl, PDGFR / FGFR family, MEK / RAF, HER2 / Neu, ubiquitin, JAK, MAP2K, ALK, PARP, transforming growth factor beta receptor (TGFβR), proteasome, Bcl-2, c-Met, VR1, VR2, VR3, c-kit, AXL, RET, Braf, DNA methyltransferase (DNMT), CDK4 / 6, and STING.

[0084] The targeted anticancer drug may be olaratumab, erlotinib, panitumumab, trastuzumab, trastuzumab emtansine, pertuzumab, cetuximab, rituximab, bevacizumab, axitinib, lenvatinib, ramucirumab, aflibercept, obinutuzumab, daratumumab, denosumab, ibrutinib, david Dasatinib, Radotinib, Nilotinib, Imatinib, Bosutinib, Galunisertib, Vactosertib, Nintedanib, Sunitinib, Sorafenib, Cabozantinib, Regorafenib, Masitinib, Semaki Semaxanib, Ceritinib, Tivozanib, Brigatinib, Vandetanib, Pazopanib, Trametinib, Temsirolimus, Dabrafenib, Dacomitinib, Apatinib, Lapatinib, Nera Neratinib, Lenalidomide, Osimertinib, Ixazomib, Olmutinib, Everolimus, Ruxolitinib, Lestaurtinib, Pacritinib, Cobimetinib, Selumetinib, Binimetinib (Binimetinib), Bortezomib, Alectinib, Crizotinib, Venetoclax, Bencentinib, Gilteritinib, Selpercatinib, Pralsetinib, Vemurafenib, Olaparib, Talazoparib It may be any one selected from the group consisting of Talazoparib, Niraparib, Rucaparib, Azacitidine, Decitabine, Guadecitabine, Gefitinib, Abemaciclib, Ribociclib, Palbociclib, and DMXAA.

[0085] The term "epidermal growth factor receptor (EGFR)" as used herein refers to a cell membrane receptor that regulates cell growth, division, survival, and death. EGFR expression is elevated in tumor tissues of various cancers. Tumor tissues with elevated EGFR expression are known to have high invasiveness, metastasis, and resistance to anticancer drugs. In one embodiment, the EGFR inhibitor may be cetuximab, trastuzumab, pertuzumab, gefitinib, erlotinib, or panitumumab.

[0086] As used herein, the term "vascular endothelial growth factor receptor (VEGFR)" refers to a cell membrane receptor for angiogenic factors that induce angiogenesis, and a VEGFR inhibitor inhibits angiogenesis to suppress tumor growth and metastasis. One embodiment of a VEGF inhibitor or a VEGFR inhibitor may be axitinib, lenvatinib, bevacizumab, ramucirumab, or aflibercept.

[0087] The term "CD20 (B lymphocyte antigen CD20)" is used herein as a target protein for treating B cell lymphoma, as it is a protein expressed on the surface of B cells. The CD20 target inhibitor may be Rituximab or Obinutuzumab.

[0088] As used herein, the term "CD38 (Cluster of differentiation 38)" refers to a protein that acts as a signal transduction receptor in immune cells and regulates cell proliferation and death, and an inhibitor that targets this protein may be daratumumab.

[0089] As used herein, the term "RNAK-L (Receptor activator of nuclear factor kappa-B ligand)" refers to the RANK-L is a receptor expressed on the surface of cells that, when activated by binding with its ligand, causes bone destruction. RANK-L inhibitors are primarily used for cancer patients suffering from bone metastasis and osteoporosis, specifically Denosumab.

[0090] As used herein, the term "BTK (Bruton's tyrosine kinase)" refers to an enzyme involved in B-cell proliferation, which, when overexpressed, can lead to hematological cancers. One embodiment of a BTK-targeted inhibitor can be ibrutinib.

[0091] The term "Bcr-abl" as used herein refers to a fusion protein that is highly expressed in patients with chronic myeloid leukemia and is known to induce abnormal proliferation of blood cells. Specifically, the inhibitor of this protein may be dasatinib, nilotinib, imatinib, or bosutinib.

[0092] The term "tumor growth factor β receptor (TGFβR)" as used herein refers to a cell membrane receptor for tumor growth factors, which regulates the growth, migration, differentiation, and death of epithelial cells and hematopoietic cells. Examples of TGFβR targeting inhibitors include, but are not limited to, Galunisertib and Vactosertib.

[0093] As used herein, "PDGFR (Platelet-derived growth factor receptor)" refers to The term "PDGFR (fibroblast growth factor)" refers to a cell membrane receptor for PDGF that is frequently expressed in cancer cells and is known to be involved in angiogenesis and regulate cancer growth, metastasis, and drug resistance. FGFR (fibroblast growth factor receptor) is a receptor for fibroblast growth factor (FGF) and regulates various biological processes, including cell growth, differentiation, and migration. Mutations in the FGFR gene are common, and such variants are commonly found in breast cancer, uterine cancer, ovarian cancer, and cervical cancer. Inhibitors targeting PDGFR or FGFR include nintedanib, sunitinib, sorafenib, cabozantinib, lenvatinib, regorafenib, masitinib, semaxanib, tivozanib, vandetanib, axitinib, and pazopanib.

[0094] As used herein, the term "MEK / RAF" refers to an intracellular signaling mediator involved in cell proliferation, cell cycle regulation, cell survival, angiogenesis, cell migration, etc., and is overactivated in cancer cells. An inhibitor targeting MEK / RAF may be trametinib or dabrafenib.

[0095] As used herein, the term "HER-2 / neu (Human epidermal growth factor receptor 2)" refers to a molecule that regulates cell proliferation by activating PI3K / AkT. It is known to be overexpressed in metastatic breast cancer, ovarian cancer, and other cancers, and induces resistance to anticancer drugs. Her2 / neu-targeting anticancer drugs may be trastuzumab, afatinib, lapatinib, or neratinib.

[0096] The term "ubiquitin" as used herein refers to a protein that maintains cellular homeostasis by binding to other proteins and inducing protein degradation by the proteasome, a proteolytic enzyme (ubiquitin-proteasome system, UPS). Abnormal expression or activity of the UPS is observed in various tumors, and inhibitors thereof exhibit anti-cancer activity. Specifically, inhibitors targeting ubiquitin or proteasomes may be lenalidomide or ixazomib.

[0097] As used herein, the term "JAK (Janus kinase)" refers to a protein upstream of STAT, a transcription factor that regulates cell proliferation, cell survival, cell migration, and immune responses. JAK inhibitors are known to reduce cell proliferation and induce cell death through inhibition of STAT activity. The JAK target inhibitor may be Ruxolitinib, Lestartinib, or Pacritinib.

[0098] As used herein, the term "MAP2K (Mitogen-activated protein kinase kinase)" refers to an intracellular signaling mediator that phosphorylates MAPK and is involved in cell proliferation, cell cycle regulation, cell survival, angiogenesis, cell migration, etc., and is overactivated in cancer cells. The MAP2K target inhibitor may be Cobimetinib, Selumetinib, Trametinib, or Binimetinib.

[0099] As used herein, the term "ALK (Anaplastic lymphoma kinase)" refers to a signal transduction mediator that promotes cell proliferation, cell migration, and angiogenesis, and suppresses cell death, and is hyperactivated in various cancer tissues. The ALK target inhibitor may be Alectinib or Crizotinib.

[0100] The term "Bcl-2" as used herein refers to a protein that inhibits cell death and is overexpressed or overactivated in various cancer tissues. An inhibitor targeting Bcl-2 may be Venetoclax.

[0101] As used herein, the term "C-Met" refers to a receptor for hepatocyte growth factor (HGF), which activates signal transduction associated with cell growth, formation, motility, survival, angiogenesis, etc. The C-Met-targeting anticancer agent may be Crizotinib or Cabozantinib.

[0102] The term "VR (Vanilloid receptor)" as used herein is also known as TRPV (Transient Receptor Peritential Vaniloid), and exists in the forms of VR1, VR2, VR3, VR4, VR5, and VR6. VRs are known to regulate the proliferation, death, migration, invasion, and angiogenesis of cancer cells at each stage of cancer progression.

[0103] As used herein, the term "C-kit" is also known as CD117, which 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 development of cancer.

[0104] As used herein, the term "AXL (Yrosin-protein kinase receptor UFO)" refers to a tyrosine kinase receptor present on the cell surface that mediates signal transduction involved in cell proliferation and survival. AXL is known to be involved in anticancer drug resistance in anticancer treatment. An example of an AXL-targeting anticancer drug is bemcentinib or gilteritinib.

[0105] The term "RET (Rearranged during transfection)" as used herein refers to a receptor that mediates signals involved in cell proliferation, cell death, and survival, and mutations in RET are known to be involved in the development of cancer. The RET target inhibitor may be selpercatinib or pralsetinib. However, the present invention is not limited to these.

[0106] The term "Braf" as used herein refers to a MAPK signaling mediator involved in cell proliferation, cell cycle regulation, cell survival, angiogenesis, cell migration, etc., and genetic mutations are observed in cancer cells. An inhibitor targeting Braf may be Vemurafenib.

[0107] The term "PARP (Poly[ADP-ribose]polymerase)" as used herein refers to a protein that recognizes damaged DNA in the nucleus, becomes activated, and then activates DNA repair-related proteins. PARP target inhibitors inhibit DNA repair in cancer cells and suppress the proliferation of cancer cells. One embodiment of the PARP target inhibitor may be Olaparib, Talazoparib, Niraparib, or Rucaparib.

[0108] As used herein, "DNA methyltransferase (DNMT)" refers to an enzyme that adds methyl groups to histone proteins that wrap around DNA, resulting in the suppression of gene expression. DMMT-targeted inhibitors exhibit anticancer activity by inhibiting hypermethylation of tumor suppressor genes and inducing their normal expression. An example of a DNMT-targeted inhibitor is azacitidine, decitabine, or guadecitabin.

[0109] As used herein, the term "cyclin-dependent kinase (CDC) 4 / 6" refers to proteins that regulate the cell cycle and promote cell growth. These proteins are overactivated during the development and progression of various malignant tumors. CDK4 / 6 targeted inhibitors exhibit anticancer activity by inhibiting the cell cycle of cancer cells, suppressing cell proliferation, and inducing cell death. The CDK4 / 6 targeted inhibitor may be Abemaciclib or Palbociclib.

[0110] As used herein, the term "STING (Stimulator of Interferon Genes)" refers to an in vivo sensor that recognizes DNA fragments from cancer cells and stimulates interferon genes to activate immune cells in the body, such as dendritic cells. STING agonists exhibit immune-enhancing effects and cancer angiogenesis-suppressing effects, and examples of STING agonists include CDNs, SB11285, and DMXAA.

[0111] The term "antibody therapeutic agent" as used herein refers to a therapeutic agent that exhibits anti-cancer effects by using an antibody that recognizes a specific protein on cancer cells as an antigen. Examples of antibody therapeutic agents include cetuximab, trastuzumab, and trastuzumab emtansine. It may be any one selected from the group consisting of, but not limited to, Emtansine, Rituximab, Ibritumomab, Tositumomab, Brentuximab, Ofatumumab, Obinutuzumab, Necitumumab, Bevacizumab, Ramucirumab, Nivolumab, Pembrolizumab, Atezolizumab, Davalumab, and Ipilimumab.

[0112] The term "immune anticancer agent" as used herein refers to an agent that inhibits the differentiation, proliferation, and activity of immune cells by inhibiting immune checkpoint proteins. It is known that the immunoanticancer agent inhibits the activity of the immune system (IL-1) and eliminates cancer cells by preventing them from evading the immune system. The immunoanticancer agent may be an antibody against any one selected from the group consisting of 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, the antibody may be any antibody selected from the group consisting of an anti-CTLA-4 antibody, an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-PD-L2 antibody, an anti-B7-H4 antibody, an anti-HVEM antibody, an anti-TIM3 antibody, an anti-GAL9 antibody, an anti-LAG3 antibody, an anti-VISTA antibody, an anti-KIR antibody, an anti-BTLA antibody, and an anti-TIGIT antibody, but is not limited to these.In one embodiment, the immunological anti-cancer agent is atezolizumab, avelumab, davalumab, nivolumab, pembrolizumab, abagovomab, adecatumumab, afutuzumab, alemtuzumab, anatumomab, mafenatoxin, or cefotaxime. Mafenatox, Mepolizumab, Blinatumomab, BMS-936559, Catumaxomab, Cemiplimab, Epacadostat, Epratuzumab, Indoximod, Inotuzumab, Inotuzumab Ozogamicin The anti-cancer drug may be any one selected from the group consisting of, but is not limited to, ozogamicin, intelumumab, ipilimumab, isatuximab, lambrolizumab, MED14736, MPDL3280A, obinutuzumab, ofatumumab, olaratumab, fidelizumab, rituximab, ticilimumab, samalizumab, and tremelimumab.

[0113] Furthermore, the immunoanticancer agent may be a fusion protein containing a membrane protein and a cytokine, specifically a fusion protein containing CD80 protein or a fragment thereof and IL-2 protein or a variant thereof, the fusion protein containing CD80 and IL-2 being the same as described above.

[0114] The term "Antibody drug conjugate (ADC)" as used herein refers to a therapeutic agent that chemically binds an antibody to a cytotoxic drug and exhibits high anticancer efficacy through targeted delivery. Examples of ADCs include gemtuzumab-ozogamicin, brentuximab-vedotin, trastuzumab-emtansine, inotuzumab-ozogamicin, and eribulin-mesylate.

[0115] The anticancer drugs include Cisplatin, Oxaliplatin, ALTIMA, Axitinib (VR1, 2, 3, PDGFR, c-kit), Galunisertib (TGFβRI), Lenvatinib (VR1, 2, 3), Ramucirumab ( VR2), Cabozatinib (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).

[0116] The Lactobacillus plantarum GB104 can be used in conjunction with the anti-cancer agent and anti-cancer vaccine, where the Lactobacillus plantarum GB104 strain and the anti-cancer agent are the same as those described above.

[0117] The term "anti-cancer vaccine" as used herein refers to an active immunotherapy that activates the immune system by administering tumor-specific antigens (TSAs) contained in cancer cells to cancer patients, thereby enhancing in vivo immune function and eliminating cancer cells. Anti-cancer vaccines include DNA vaccines, peptide vaccines, and cellular vaccines depending on the type of antigen and antigen delivery method. Currently, cellular vaccines and DNA vaccines, which are developed by introducing antigens, are the most commonly developed.

[0118] In one embodiment, the Lactobacillus plantarum GB104 strain can be used in combination with two or more different anticancer agents (e.g., a first anticancer agent and a second anticancer agent). For example, the two anticancer agents (e.g., the first anticancer agent and the 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, or a chemical anticancer agent and an immunological anticancer agent. Furthermore, they can 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, or a targeted anticancer agent and an immunological anticancer agent. Furthermore, they can be an anticancer virus and an antibody therapeutic agent, an anticancer virus and a cell therapeutic agent, or an anticancer virus and an immunological anticancer agent. Furthermore, they can be an antibody therapeutic agent and a cell therapeutic agent, or an antibody therapeutic agent and an immunological anticancer agent. In one embodiment of the present invention, the two anti-cancer agents can be an immunological anti-cancer agent and a chemical anti-cancer agent, an immunological anti-cancer agent and a targeted anti-cancer agent, or an immunological anti-cancer agent and an antibody therapeutic agent.

[0119] In one embodiment, the first anticancer agent administered in combination with the Lactobacillus plantarum GB104 strain is a fusion protein comprising a CD80 protein or a fragment thereof and an IL-2 protein or a variant thereof, and the second anticancer agent may be one or more selected from the group consisting of 5-fluorouracil, anti-PD-1, bevacizumab, cetuximab, and regorafenib.

[0120] In one embodiment, the first and second anti-cancer agents may be co-administered simultaneously, sequentially or in reverse order.

[0121] As used herein, the terms "combination therapy" or "co-administration" or "in combination" refer to any form of simultaneous or combined therapy using at least two separate therapeutic agents. The components of the combination therapy can be administered simultaneously, sequentially, or in any order. The components can be administered in different doses, at different administration frequencies, or via different routes as appropriate.

[0122] Specifically, the combined administration can involve administering a first anticancer agent and a second anticancer agent simultaneously, or administering the first anticancer agent followed by the second anticancer agent. A combination therapy according to the present invention can be defined as providing a synergistic effect if its efficacy, measured, for example, through the degree of response, response rate, time to disease progression, or survival time, is therapeutically superior to the efficacy obtained by administering one or the other of the components of the combination therapy at a conventional dose. For example, the efficacy of a combination therapy is synergistic if its efficacy is therapeutically superior to the efficacy obtained by using each component alone. In particular, the efficacy of a combination therapy can be defined as being synergistic when it does not adversely affect one or more of the degree of response, response rate, time to disease progression, and survival data, particularly the duration of response, and when each component is used at a conventional dose. A synergistic effect is considered to exist if the side effects of concern are reduced / few or fewer than those that occur when the first and second anticancer agents are administered together and the usual doses of the first and second anticancer agents can be reduced.

[0123] The term "administered simultaneously" as used herein is not particularly limited and means that the components of the combination therapy are administered substantially simultaneously, for example, as an admixture or in immediately subsequent order.

[0124] The term "sequentially administered" as used herein is not particularly limited and means that the components of a combination therapy are not administered simultaneously, but rather are administered one after the other or in groups with a specific time interval between administrations. The time intervals may be the same or different between each administration of the components of the combination therapy, and may be selected from the ranges of, for example, 2 minutes to 96 hours, 1 day to 7 days, or 1 week, 2 weeks, or 3 weeks. Generally, the time interval between administrations may range from a few minutes to several hours, for example, 2 minutes to 72 hours, 30 minutes to 24 hours, or 1 to 12 hours. Further examples include time intervals ranging from 24 to 96 hours, 12 to 36 hours, 8 to 24 hours, and 6 to 12 hours.

[0125] In one embodiment, the first anticancer agent and the second anticancer agent can be administered via independent routes. Each active ingredient can be independently administered at an appropriate dosage by a skilled artisan. Specifically, the first anticancer agent and the second anticancer agent can be administered intratumorally, intraarterially, intravenously, intravascularly, intrapleurally, intraperitoneally, intratracheally, intradurally, intramuscularly, endoscopically, intralesionally, intradermally, subcutaneously, regionally, stereotactically, or orally by direct injection or perfusion. 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.

[0126] In one embodiment, the second active substance may comprise a first anti-cancer agent, or a first and a second anti-cancer agent, alone as the active ingredient, or may comprise one or more pharmaceutically acceptable carriers, excipients, or diluents.

[0127] In one embodiment, the anticancer agent can be prepared as a separate formulation for administration in combination with the composition of the present invention. The anticancer agent can be prepared as an injectable solution containing the anticancer agent, preferably a formulation for administration by direct injection or perfusion, such as intratumoral, intraarterial, intravenous, intravascular, intrapleural, intraperitoneal, intratracheal, intradural, intramuscular, endoscopic, intralesional, percutaneous, subcutaneous, regional, stereotactic, or oral administration, but is not limited thereto.

[0128] In this specification, the term "containing as an active ingredient" means that a Lactobacillus strain, a disrupted solution of the strain, a culture solution, or an extract of the culture solution is added, and various components are added as accessory components for drug delivery, stabilization, etc., and this also includes formulation in various forms.

[0129] In one embodiment, the first and second active agents can be co-administered simultaneously, sequentially or in reverse order.

[0130] The "strain," "first active substance," "second active substance," "first anticancer agent," "second anticancer agent," and "combined administration" are as described above.

[0131] Specifically, the combined administration involves administering a Lactobacillus plantarum strain and an anticancer drug at the same time. Alternatively, the Lactobacillus plantarum strain may be administered first, or an anticancer drug may be administered after the administration of the Lactobacillus plantarum strain. The anticancer drugs may include a first anticancer drug and a second anticancer drug that are different from each other. More specifically, the first anticancer drug may be an immunosuppressant. A combination therapy according to the present invention may be defined as providing a synergistic effect if its efficacy, measured, for example, through the degree of response, response rate, time to disease progression, or survival time, is therapeutically superior to the efficacy obtained by administering one or the other of the components of the combination therapy at a conventional dose. For example, the efficacy of a combination therapy is synergistic if its efficacy is therapeutically superior to the efficacy obtained by administering each component alone. In particular, a synergistic effect is considered to exist if the degree of response, response rate, time to disease progression, and survival data are not adversely affected, particularly the duration of response, and if problematic side effects are reduced or eliminated compared to those occurring when each component is administered at a conventional dose, and if the conventional doses of the Lactobacillus plantarum strain and the anticancer drug can be reduced.

[0132] In one embodiment, the synergistic effect of the combined administration may be a reduction in the dosage or frequency of administration of an existing anticancer drug. Specifically, when the Lactobacillus plantarum GB104 strain is combined with an immunological anticancer drug (e.g., a fusion protein comprising a CD80 protein or a fragment thereof and an IL-2 protein or a variant thereof), the combined administration may exhibit the same or similar anticancer activity at a lower dose or with fewer administrations than the immunological anticancer drug administered alone.

[0133] In one embodiment, when the administration interval of the immunosuppressant alone was one day, the anticancer activity was the same or similar when the administration interval of the Lactobacillus plantarum GB104 strain and the immunosuppressant was five to ten days (e.g., seven days) in combination. Therefore, the coadministration of Lactobacillus plantarum GB104 has a synergistic effect that can reduce the usual dose and frequency of the immunosuppressant.

[0134] In one embodiment, when Lactobacillus plantarum GB104 is administered in combination with an immunosuppressive agent (e.g., a fusion protein comprising a CD80 protein or a fragment thereof and an IL-2 protein or a variant thereof) and a chemoanticancer agent, the anticancer activity can be the same or similar at a lower dose or with fewer administrations than the immunosuppressive agent or chemoanticancer agent administered alone. In one embodiment, the anticancer activity of the immunosuppressive agent or chemoanticancer agent administered alone at an interval of one day was comparable to the anticancer activity of the Lactobacillus plantarum GB104, immunosuppressive agent, and chemoanticancer agent administered in combination at an interval of 5 to 10 days (e.g., 7 days). Therefore, the combined administration of Lactobacillus plantarum GB104 has a synergistic effect that allows the usual dose and frequency of administration of the immunosuppressive agent and / or chemoanticancer agent to be reduced.

[0135] In one embodiment, the first active substance and the second active substance can be administered via independent routes. Each active ingredient can be independently administered at an appropriate dosage by a skilled artisan. Specifically, the Lactobacillus plantarum GB104 and the anticancer agent can be administered intratumorally, intraarterially, intravenously, intravascularly, intrapleurally, intraperitoneally, intratracheally, intradurally, intramuscularly, endoscopically, intralesionally, intradermally, subcutaneously, regionally, stereotactically, or orally by direct injection or perfusion. 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 this is not limited thereto.

[0136] In one embodiment, the first active substance and the second active substance may be administered to an individual at intervals of 1 to 10 days. More specifically, the first active substance may be administered to an individual at intervals of 1 to 3 days, and the second active substance may be administered to an individual at intervals of 5 to 10 days. In one embodiment, the first active agent is administered to the individual at one day intervals and the second active agent is administered to the individual at seven day intervals.

[0137] Another aspect is to provide a pharmaceutical composition for preventing or treating cancer, which comprises as active ingredients a first active substance comprising one or more selected from the group consisting of a Lactobacillus plantarum strain, a culture of the strain, a disrupted product of the strain, or an extract of the strain, culture, or disrupted product, and a second active substance comprising an immunological anticancer agent that is a first anticancer agent, or which comprises the first active substance as an active ingredient and is administered in combination with the second active substance.

[0138] The "strain," "first active substance," "anticancer agent," "second active substance," "anticancer activity," and "combined administration" are as defined above.

[0139] In one embodiment, the pharmaceutical composition can be administered to mammals, including humans, via various routes. The administration route can be any commonly used route, such as oral, intradermal, intravenous, intramuscular, or subcutaneous, and preferably oral.

[0140] In one embodiment, the formulation comprises a first oral formulation comprising a first active agent and a second oral formulation comprising a second active agent, wherein said first and second oral formulations can be administered orally.

[0141] The term "prevention" as used herein can refer to any act of inhibiting or delaying the onset of a disease state in an individual by administering a pharmaceutical composition according to an embodiment.

[0142] The term "treatment" as used herein can refer to any action in which the symptoms of an individual's disease state are ameliorated or beneficially altered by administration of a pharmaceutical composition according to one embodiment.

[0143] According to one embodiment, the composition may contain 0.001 wt% to 80 wt% of the Lactobacillus plantarum strain based on the total weight of the composition. The dosage of the Lactobacillus plantarum strain may be 0.01 mg to 10,000 mg, 0.1 mg to 1,000 mg, 1 mg to 100 mg, 0.01 mg to 1,000 mg, 0.01 mg to 100 mg, 0.01 mg to 10 mg, or 0.01 mg to 1 mg. The strain is contained in the composition at a therapeutically or nutritionally effective amount, for example, when the strain is 10 3 ~10 16 CFU / g, 10 3 ~10 15 CFU / g, 10 3 ~10 14 CFU / g, 10 3 ~10 13 CFU / g, 10 3 ~10 12 CFU / g, 10 4 ~10 16 CFU / g, 10 4 ~10 15 CFU / g, 10 4 ~10 14 CFU / g, 10 4 ~10 13 CFU / g, 10 4 ~10 12 CFU / g, 10 5 ~10 16 CFU / g, 10 5 ~10 15 CFU / g, 10 5 ~10 14 CFU / g, 10 5 ~10 13 CFU / g, 10 5 ~10 12 CFU / g, 10 6 ~10 13 CFU / g, 10 6 ~10 12 CFU / g, 10 7 ~10 13 CFU / g, 10 7 ~10 12 CFU / g, 10 8 ~10 13 CFU / g or 10 8 ~1012 The composition may contain 1×10 CFU / g or a culture of the same number of live or dead bacteria. 3 ~1×10 16 CFU / g of live or killed bacteria can be administered in one or several doses. However, the dosage can be formulated in various ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate and reaction sensitivity, and those skilled in the art can appropriately control the dosage taking these factors into consideration. The number of administrations can be one or two or more times within the range of clinically acceptable side effects, and the administration site can be one or more sites. For animals other than humans, the dosage can be determined in one or several doses per kg (body weight) The dosage may be the same as that for humans per 100 mg / kg body weight, or may be calculated by converting the dosage into the above-mentioned dosage based on, for example, the volume ratio (e.g., average value) of organs (e.g., heart) between the target animal and humans. Possible routes of administration include oral, sublingual, parenteral (e.g., subcutaneous, intramuscular, intraarterial, intraperitoneal, intradural, or intravenous), rectal, topical (including transdermal), inhalation, and injection, or insertion of an implantable device or substance. Target animals for treatment according to one embodiment include humans and other mammalian species, specifically humans, monkeys, mice, rats, rabbits, sheep, cattle, dogs, horses, pigs, etc. According to one embodiment, the composition contains a killed dried strain of bacteria, and can be administered at a dose of 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 per dose, once to three times daily.

[0144] The term "therapeutically effective amount" as used herein refers to an amount of an anti-cancer agent for the methods and uses of the present invention, or a pharmaceutical composition containing an anti-cancer agent for the methods and uses of the present invention, that elicits the biological or medical response or desired therapeutic effect in a patient that a researcher, physician, or other clinician seeks to achieve. A therapeutically effective amount of an anti-cancer agent can vary depending on factors such as the disease state, age, sex, and weight of the individual, and the ability of the anti-cancer agent to elicit a desired response in the individual. A therapeutically effective amount is one in which the therapeutically beneficial effects outweigh any toxic or detrimental effects.

[0145] The pharmaceutical composition according to one embodiment may contain pharmaceutically acceptable carriers and / or additives, such as sterile water, physiological saline, conventional buffers (such as phosphate, citric acid, and other organic acids), stabilizers, salts, antioxidants (such as ascorbic acid), surfactants, suspending agents, isotonicity agents, or preservatives. For topical administration, the composition may be combined with organic materials such as biopolymers, inorganic materials such as hydroxyapatite, specifically collagen matrices, polylactic acid polymers or copolymers, polyethylene glycol polymers or copolymers, and chemical derivatives thereof.

[0146] When the pharmaceutical composition according to one embodiment is prepared in a formulation suitable for injection, the Lactobacillus bacteria may be dissolved or dispersed in a pharmaceutically acceptable carrier, or may be frozen in a solution in which they are dissolved or dispersed.

[0147] A pharmaceutical composition according to one embodiment may appropriately contain suspending agents, solubilizers, stabilizers, isotonicity adjusting agents, preservatives, anti-adsorption agents, surfactants, diluents, excipients, pH adjusters, soothing agents, buffers, reducing agents, antioxidants, and the like, as needed depending on the administration method or formulation. Pharmaceutically acceptable carriers and formulations suitable for the present invention, including those exemplified above, are described in detail in Remington's Pharmaceutical Sciences, 19th ed., 1995. A pharmaceutical composition according to one embodiment may be prepared in unit dose form by formulating the composition with pharmaceutically acceptable carriers and / or excipients, or by filling a large-volume container, according to a method readily practiced by a person skilled in the art to which the present invention pertains. In this case, the formulation 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.

[0148] The pharmaceutical composition is administered in a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to any medical treatment. The effective dose level can be determined based on factors including the type and severity of the patient's disease, the activity of the drug, sensitivity to the drug, the time of administration, the route of administration and excretion rate, the duration of treatment, concurrently used drugs, and other factors well known in the medical field. The composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or multiple times. Taking all of the above factors into consideration, it is desirable to obtain maximum effect with minimum amount without side effects. It is important to administer an amount that will achieve the desired effect, which can be readily determined by one skilled in the art.

[0149] In one embodiment, the anti-cancer activity of the first active agent may be more effective when administered in combination with a second active agent.

[0150] In one embodiment, when the Lactobacillus plantarum GB104 strain was administered in combination with the anticancer drug 5-fluorouracil in a tumor animal model implanted with the mouse colon cancer cell line MC-38, it exhibited enhanced anticancer effects compared to administration of the Lactobacillus plantarum GB104 strain alone. Specifically, the effect of inhibiting the growth and progression of colon cancer tumors was confirmed. On average, tumor volume was reduced by 10% to 90% in the experimental group administered with the combination of GB104 and the anticancer drug compared to the control group.

[0151] In one embodiment, when the Lactobacillus plantarum GB104 strain is administered in combination with an anticancer drug, the colon cancer tumor volume may be reduced to a level of 90% or less, 80% or less, 70% or less, 67% or less, 10-90%, 10-80%, 10-70%, 10-67%, 20-90%, 20-80%, 20-70%, 20-67%, 30-90%, 30-80%, 30-70%, 30-67%, 40-90%, 40-80%, 40-70%, 40-67%, 50-90%, 50-80%, 50-70%, 50-67%, or 60-67%, based on 100% colon cancer tumor volume in a negative control group that was not administered the strain or anticancer drug.

[0152] In one embodiment, when the Lactobacillus plantarum GB104 strain was administered in combination with the anti-PD-1 immunotherapy agent in a tumor animal model implanted with the mouse colon cancer cell line MC-38, it exhibited enhanced anti-cancer effects compared to administration of the Lactobacillus plantarum GB104 strain alone. Specifically, the effect of inhibiting the growth and progression of colon cancer tumors was confirmed. On average, tumor volume was reduced by 10% to 90% in the experimental group administered with the GB104 immunotherapy agent in combination compared to the control group.

[0153] In one embodiment, when the Lactobacillus plantarum GB104 strain was administered in combination with immuno-anticancer agents GI-101 or GI-102, which are fusion proteins comprising a CD80 fragment and an IL-2 protein variant, to a tumor animal model implanted with the mouse colon cancer cell line MC-38, the anticancer effects were enhanced compared to administration of the Lactobacillus plantarum GB104 strain alone. Specifically, the effect of inhibiting the growth and progression of colon cancer tumors was confirmed. On average, tumor volume was reduced by 10% to 90% in the experimental group administered with the GB104 immuno-anticancer agent in combination compared to the control group.

[0154] In one embodiment, when the Lactobacillus plantarum GB104 strain is administered in combination with an immunological anticancer agent, the colon cancer tumor volume may be reduced to a level of 90% or less, 80% or less, 70% or less, 67% or less, 10-90%, 10-80%, 10-70%, 10-67%, 20-90%, 20-80%, 20-70%, 20-67%, 30-90%, 30-80%, 30-70%, 30-67%, 40-90%, 40-80%, 40-70%, 40-67%, 50-90%, 50-80%, 50-70%, 50-67%, or 60-67%, based on 100% colon cancer tumor volume in a negative control group that was not administered the strain or anticancer agent.

[0155] In one embodiment, when the Lactobacillus plantarum GB104 strain was administered in combination with the targeted anticancer drugs Cetuximab, Bevacizumab, or Regorafenib in a tumor animal model implanted with the mouse colon cancer cell line MC-38, an enhanced anticancer effect was observed compared to when the Lactobacillus plantarum GB104 strain was administered alone. Specifically, the effect of inhibiting the growth and progression of colon cancer tumors was confirmed. On average, the control Compared to the control group, the experimental group receiving GB104 in combination with a targeted cancer drug showed a 10% to 90% reduction in tumor volume.

[0156] In one embodiment, when the Lactobacillus plantarum GB104 strain is administered in combination with a targeted anticancer drug, the Lactobacillus plantarum GB104 strain may have an activity that reduces colorectal cancer tumor volume to a level of 90% or less, 80% or less, 70% or less, 67% or less, 10-90%, 10-80%, 10-70%, 10-67%, 20-90%, 20-80%, 20-70%, 20-67%, 30-90%, 30-80%, 30-70%, 30-67%, 40-90%, 40-80%, 40-70%, 40-67%, 50-90%, 50-80%, 50-70%, 50-67%, or 60-67%, based on a colorectal cancer tumor volume of 100% in a negative control group that was not administered the strain or anticancer drug.

[0157] In one embodiment, when Lactobacillus plantarum GB104 strain, an immunological anticancer agent, and a chemical anticancer agent are administered in combination, the colon cancer tumor volume may be reduced to a level of 90% or less, 80% or less, 70% or less, 67% or less, 10-90%, 10-80%, 10-70%, 10-67%, 20-90%, 20-80%, 20-70%, 20-67%, 30-90%, 30-80%, 30-70%, 30-67%, 40-90%, 40-80%, 40-70%, 40-67%, 50-90%, 50-80%, 50-70%, 50-67%, or 60-67%, based on 100% colon cancer tumor volume in a negative control group that was not administered the strain or anticancer agent.

[0158] In one embodiment, a triple combination of the Lactobacillus plantarum GB104 strain, the immunological anticancer agent GI-101, and the chemical anticancer agent 5-FU was administered to a tumor animal model implanted with the mouse colon cancer cell line MC-38. The triple combination showed enhanced anticancer effects compared to administration of the Lactobacillus plantarum GB104 strain alone or administration of the Lactobacillus plantarum GB104 strain and the immunological anticancer agent GI-101. Specifically, the effect of inhibiting the growth and progression of colon cancer tumors was confirmed. On average, tumor volume was reduced by 10% to 90% in the experimental group receiving the triple combination of GB104, the immunological anticancer agent, and the chemical anticancer agent compared to the control group.

[0159] Another aspect is to provide a health functional food for preventing or ameliorating cancer, which comprises a first active substance comprising one or more selected from the group consisting of a Lactobacillus plantarum strain, a culture of said strain, a disrupted product of said strain, or extracts of said strain, culture, and disrupted product, and is administered in combination with a second active substance comprising an immunological anticancer agent as the first anticancer agent.

[0160] Another aspect is to provide a food composition for preventing or ameliorating cancer, which comprises as active ingredients a first active substance comprising one or more selected from the group consisting of a Lactobacillus plantarum strain, a culture of the strain, a disrupted product of the strain, or an extract of the strain, culture, and disrupted product, and a second active substance comprising an immunological anticancer agent that is a first anticancer agent, or which comprises the first active substance as an active ingredient and is administered in combination with the second active substance.

[0161] The "strain," "first active substance," "second active substance," "first anticancer agent," "second anticancer agent," "anticancer activity," and "combined administration" are as defined above.

[0162] 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, and preferably oral administration.

[0163] In one embodiment, the formulation comprises a first oral formulation comprising a first active agent and a second oral formulation comprising a second active agent, wherein said first and second oral formulations can be administered orally.

[0164] In one embodiment, the health functional food may further comprise a nutrient-acceptable carrier.

[0165] The term "food-based acceptable" as used herein means that the compound exhibits the property of being non-toxic to cells or humans exposed to the compound.

[0166] The term "improvement" as used herein can refer to any action that at least reduces a parameter related to the condition being treated, such as the severity of symptoms. In this case, the health functional food can be used to prevent or improve cancer before or after the onset of the disease, simultaneously with or separately from a therapeutic drug.

[0167] In the health functional foods, the active ingredient may be added directly to the food or may be used in combination with other foods or food ingredients, or may be used appropriately according to conventional methods. The amount of the active ingredient to be mixed can be determined appropriately depending on the intended use (prevention or improvement). Generally, in the production of foods or beverages, the health functional foods can be added in an amount of about 15% by weight or less, more specifically about 10% by weight or less, based on the raw materials. However, in the case of long-term intake for health and hygiene purposes or health regulation purposes, the amount may be less than the above range.

[0168] The health functional food may be formulated into one selected from the group consisting of tablets, pills, powders, granules, powders, capsules, and liquid preparations, further comprising one or more of a carrier, a diluent, an excipient, and an additive. Foods to which the compound according to one embodiment can be added include various foods, powders, granules, tablets, capsules, syrups, beverages, gum, tea, vitamin complexes, health functional foods, etc.

[0169] Specific examples of the carrier, excipient, diluent, and additive may be at least one selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium phosphate, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, methylcellulose, water, gum syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0170] In addition to the active ingredient, the health functional food may contain other essential ingredients without particular limitation. For example, like ordinary beverages, it may contain various flavorings or natural carbohydrates as additional ingredients. Examples of the natural carbohydrates include conventional sugars such as monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), polysaccharides (e.g., dextrin, cyclodextrin, etc.), and sugar alcohols (e.g., xylitol, sorbitol, erythritol, etc.). Other flavorings that can be advantageously used include natural flavorings (thaumatin, stevia extract (e.g., rebaudioside A, glycylrhizin, etc.)) and synthetic flavorings (saccharin, aspartame, etc.). The proportion of the natural carbohydrates can be appropriately determined by the selection of a person skilled in the art.

[0171] In addition to the above, the health functional food according to one embodiment contains various nutrients, vitamins, minerals (electrolytes), flavorings such as synthetic flavorings and natural flavorings, colorings and enhancers (cheese, chocolate, etc.). It may contain, for example, pectinic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated drinks, etc. Such ingredients may be used independently or in combination, and the proportions of such additives may also be appropriately selected by those skilled in the art.

[0172] The health functional food may be provided in a mixture with a conventionally known health functional food for preventing or improving cancer or other existing health functional foods, and the health functional food for preventing or improving cancer may be a conventionally known health functional food for preventing or improving metabolic diseases, an existing health functional food, or a newly developed health functional food.

[0173] When the health functional food contains other health functional foods that have cancer prevention or improvement effects, it is important to mix them in an amount that can achieve the maximum effect with the minimum amount without side effects, which can be easily determined by a person skilled in the art.

[0174] The food compositions for preventing or ameliorating cancer include all forms such as functional foods, nutritional supplements, health foods, and food additives, and the food compositions of this type can be prepared in various forms according to conventional methods known in the art.

[0175] The compositions herein can be considered food supplements. Food supplements, also known as dietary or nutritional supplements, can be considered another specific pharmaceutical product. They are prepared to supplement the diet and provide nutrients or beneficial ingredients that are not available or not available in sufficient amounts through a normal diet. Most food supplements are considered foods, but they are sometimes considered drugs, natural health products, or nutraceutical products. Within the meaning of the present invention, food supplements include nutraceuticals. Food supplements are usually sold over the counter without a prescription. When food supplements take the form of pills or capsules, they contain the same excipients as pharmaceuticals. However, food supplements can also take the form of food fortified with certain nutrients (e.g., infant formula). Thus, in certain embodiments, the compositions of the present invention are food supplements.

[0176] The compositions according to the present invention may be administered as is, or may be mixed with an appropriate edible liquid or solid, or may be in the form of tablets, pills, capsules, lozenges, granules, powders, suspensions, sachets, syrups, or freeze-dried in unit dose form, or may be in the form of monodoses of freeze-dried compositions to be mixed in a separate liquid container provided together before administration.

[0177] The compositions of the present invention may be included in a variety of edible foods and food products, such as milk products for infants. The term "edible product" as used herein is used in a broad sense to include any form of product that can be ingested by an animal in any form. The term "food product" is understood as an edible product that provides nutritional support to the body. Particularly interesting foods are food supplements and infant formulas. The food preferably comprises a carrier material such as oatmeal gruel, lactic acid fermented foods, resistant starch, dietary fibers, carbohydrates, proteins, and glycosylated proteins. In a particular embodiment, the bacterial cells of the invention are homogenized with other ingredients such as cereals or powdered milk to form infant formulas.

[0178] Another aspect provides a kit for preventing or treating cancer, which comprises, as active ingredients, a first active substance comprising one or more selected from the group consisting of a Lactobacillus plantarum strain, a culture of the strain, a disrupted product of the strain, or an extract of the strain, culture, or disrupted product, and a second active substance comprising an immunological anticancer agent that is a first anticancer agent, or which comprises, as an active ingredient, the first active substance and the second active substance administered in combination.

[0179] Another aspect provides a method for delivering a drug into an individual, comprising the step of administering to an individual in need thereof a composition containing as active ingredients a first active substance comprising one or more selected from the group consisting of a Lactobacillus plantarum strain, a culture of the strain, a disruptant of the strain, or an extract of the strain, culture, or disruptant, and a second active substance comprising an immunological anti-cancer agent that is a first anti-cancer agent, or a composition containing the first active substance as an active ingredient and the second active substance in combination.

[0180] Another aspect provides a method for preventing or treating cancer, comprising the step of administering to an individual in need thereof a composition containing as active ingredients a first active substance comprising one or more selected from the group consisting of a Lactobacillus plantarum strain, a culture of the strain, a disruptant of the strain, or an extract of the strain, culture, or disruptant, and a second active substance comprising an immunological anti-cancer agent that is a first anti-cancer agent, or a composition containing the first active substance as an active ingredient and the second active substance administered in combination.

[0181] The "strain," "first active substance," "second active substance," "first anticancer agent," "second anticancer agent," "anticancer activity," and "combined administration" are as defined above.

[0182] The individual may be an individual suffering from cancer. Furthermore, the individual may be a mammal, preferably a human.

[0183] The administration route, dosage, and frequency of the Lactobacillus plantarum GB104 strain and anticancer drug can be administered to a subject in various ways and amounts depending on the patient's condition and the presence or absence of side effects, and the optimal administration method, dosage, and frequency of administration can be selected within an appropriate range by a person skilled in the art. Furthermore, in addition to the active ingredient, the Lactobacillus plantarum GB104 strain and anticancer drug can be administered in combination with other drugs (e.g., the above-mentioned anticancer drugs) or physiologically active substances known to be effective in treating cancer diseases, or can be formulated in the form of a combination preparation with other drugs. [Effects of the Invention]

[0184] The Lactobacillus plantarum strain or its culture medium can inhibit the proliferation of cancer cells and induce their death, and also has the potential to inhibit the proliferation of cancer cells and induce their death when administered in combination with anticancer drugs. When used in combination, they exhibit a synergistic effect in inhibiting the proliferation of cancer cells and tumors, and can be usefully used in the form of a pharmaceutical composition or a health functional food for the prevention, treatment, or amelioration of cancer. [Brief explanation of the drawings]

[0185] [Figure 1] FIG. 1 is a graph showing the results of analyzing the cell viability after treating the human colon cancer cell strain HCT116 with culture supernatants of strains different from GB104. [Figure 2] FIG. 2 is a graph showing the results of examining the cell cycle after treating the human colon cancer cell line HCT116 with the culture supernatants of GB104 and a control strain (WCFS1). [Figure 3] FIG. 3 is a graph showing the cancer cell death effect after treating the human colon cancer cell line HCT116 or the human breast cancer cell line MDA-MB-231 with the culture supernatant of GB104 or a control strain (WCFS1). [Figure 4] FIG. 4 is a graph showing the tumorigenesis inhibitory effect of treating the mouse MC-38 colon cancer cell line, LLC1 lung cancer cell line, and 4T1 breast cancer cell line with the GB104 culture supernatant. [Figure 5]FIG. 5 is a graph showing the results of analyzing the synergistic effect by measuring tumor volume after combined administration of GB104 and 5-FU in an allografted MC-38 mouse colon carcinoma model. [Figure 6] FIG. 6 is a graph showing the results of comparative analysis of tumor volume measured after single administration and combined administration of GB104 and 5-FU in an allografted MC-38 mouse colon carcinoma model. [Figure 7] FIG. 7 is a graph showing the results of analyzing the synergistic effect by measuring tumor volume after co-administration of GB104 and anti-PD-1 in an allografted MC-38 mouse colon carcinoma model. [Figure 8] FIG. 8 is a graph showing the results of a comparative analysis of tumor volume measured after single and combined administration of GB104 and anti-PD-1 in an allografted MC-38 mouse colon carcinoma model. [Figure 9] FIG. 9 is a graph showing the results of analyzing synergistic effects by measuring tumor volume after combined treatment with GB104 and bevacisumab in a xenografted HT-29 mouse colon carcinoma model. [Figure 10] FIG. 10 is a graph showing the results of a comparative analysis of tumor volume measured after single administration and combined administration of GB104 and Bevacizumab in a xenografted HT-29 mouse colon carcinoma model. [Figure 11] FIG. 11 is a graph showing the results of analyzing synergistic effects by measuring tumor volume after combined treatment with GB104 and Cetuximab in a xenografted HT-29 mouse colon carcinoma model. [Figure 12] FIG. 12 is a graph showing the results of a comparative analysis of tumor volume measured after single administration and combined administration of GB104 and Cetuximab in a xenografted HT-29 mouse colon carcinoma model. [Figure 13] FIG. 13 is a graph showing the results of analyzing the synergistic effect by measuring tumor volume and size after combined administration of GB104 and Regorafenib in an allografted MC-38 mouse colon carcinoma model. [Figure 14]FIG. 14 is a graph showing the results of a comparative analysis of tumor volume measured after single administration and combined administration of GB104 and Regorafenib in an allografted MC-38 mouse colon carcinoma model. [Figure 15] FIG. 15 is a graph showing the results of analyzing synergistic effects by measuring tumor volume and weight after combined treatment with GB104 and GI-101 in an allografted MC-38 mouse colon carcinoma model. [Figure 16] FIG. 16 is a graph showing the results of comparative analysis of tumor volume measured after single administration and combined administration of GB104 and GI-101 in an allografted MC-38 mouse colon carcinoma model. [Figure 17] FIG. 17 is a graph showing the results of analyzing synergistic effects by measuring tumor volume and weight after combined treatment with GB104 and GI-102 in an allografted MC-38 mouse colon carcinoma model. [Figure 18] FIG. 18 is a graph showing the results of comparative analysis of tumor volume measured after single administration and combined administration of GB104 and GI-102 in an allografted MC-38 mouse colon carcinoma model. [Figure 19] FIG. 19 is a graph showing the results of analyzing synergistic effects by measuring tumor volume after triple combination treatment of GB104, GI-101, and 5-FU in an allografted MC-38 mouse colon carcinoma model. [Figure 20] FIG. 20 is a graph showing the results of a comparative analysis of tumor volume measured after single administration and combined administration of GB104, GI-101, and 5-FU in an allografted MC-38 mouse colon carcinoma model. DETAILED DESCRIPTION OF THE INVENTION

[0186] Preferred embodiments are presented below to aid in understanding the present invention. However, the following embodiments are provided to facilitate understanding of the present invention, and the contents of the present invention are not limited to the following embodiments. Various modifications can be made to the embodiments, and the embodiments are not limited to the embodiments disclosed below, and can be implemented in various forms.

[0187] Example 1. Isolation and identification of Lactobacillus plantarum GB104 strain

[0188] The isolation and identification of Lactobacillus plantarum strain GB104 was performed according to the methods described in Korean Patent Application Nos. 10-2020-0186738 and 10-2022-0080567, both of which are incorporated herein by reference in their entireties.

[0189] Briefly, Lactobacillus plantarum GB104 was isolated from vaginal samples of healthy women who visited a hospital for a health checkup. First, vaginal samples were collected with a cotton swab and pre-inoculated onto Rogosa SL (MRS) plates. They were then cultured in an anaerobic chamber at 37°C for 48 hours. Once bacterial colonies had grown, single colonies were subcultured onto new MRS plates for pure isolation. After isolation, the strains were cultured in MRS medium. Next, the Lactobacillus plantarum GB104 strain, which exhibited low levels of adipocyte accumulation inhibition and cytotoxicity, was selected from the cultured strains. To identify the selected Lactobacillus plantarum GB104 strain, the 16S rRNA gene sequence obtained by PCR using primers targeting the 16S rRNA gene was analyzed by Sanger sequencing. The 16S rRNA sequence of Lactobacillus plantarum GB104 is shown in SEQ ID NO: 1. The inventors named GB104 "Lactobacillus plantarum GB104" (deposit number: KCTC 14107BP) and deposited it with the Korean Collection for Type Cultures (KCTC) at the Korea Institute of Bioscience and Biotechnology on January 14, 2020. Furthermore, the strain name of Lactobacillus plantarum was changed to Lactiplantibacillus plantarum. In the following embodiments, the changed strain names of existing strains are used interchangeably.

[0190] Experimental Example 1: Confirmation of cancer cell viability and cancer cell killing effect by treatment with GB104 strain culture supernatant 1.1. Confirmation of cancer cell viability by treatment with GB104 culture supernatant

[0191] In this experiment, various L. plantarum culture supernatants, including the L. plantarum GB104 strain, were treated with human colon cancer cell lines, and cell viability was screened by MTT assay.

[0192] 2x10 human colon cancer cell line HCT116 cells were placed in a 96-well plate. 3 After 24 hours of incubation, cells were dispensed into each well and treated with 10% culture supernatant from various L. plantarum strains. Cultures were then incubated at 37°C and 5% CO2 for 72 hours. Culture supernatants were prepared by culturing L. plantarum strains in MRS medium, centrifuging the cultures, and filtering the supernatant through a 0.22 μm filter. Cell Proliferation Kit I (MTT) (Roche) was used to prepare the MTT solution at 0.5 mg / mL. After incubation for an additional 4 hours, purple formazan crystals formed by metabolic activity in live cells were dissolved in solubilization solution, and the absorbance was measured at 570 nm. The results are shown in Figure 1. FIG. 1 is a graph showing the results of analyzing the cell viability after treating the human colon cancer cell strain HCT116 with culture supernatants of strains different from GB104. As shown in Figure 1, various L. Plantarum culture supernatants did not show the same inhibitory effect on cancer cell proliferation. In particular, GB104 culture supernatant showed an effect of reducing cancer cell viability by approximately 95%, confirming that it was the most effective in inhibiting cancer cell proliferation.

[0193] 1.2. Confirmation of viability of human cancer cell lines by treatment with GB104 culture supernatant

[0194] In this experiment, various human endometrial cancer cell lines were treated with the culture supernatant of L. plantarum GB104 strain, and the cell viability was screened by MTT assay.

[0195] 0.8-1.5 x 10 human cancer cell lines in a 96-well plate 4 After culturing for 24 hours, the culture supernatant of L. plantarum GB104 strain was treated at a concentration of 10% with colon cancer, lung cancer, stomach cancer, breast cancer, and liver cancer cell lines, and at a concentration of 20% with kidney cancer and bladder cancer cell lines. After culturing for 48 hours at 37°C and 5% CO2, cell viability was measured in the same manner as in Experimental Example 1.1. The results are shown in Tables 1 and 2.

[0196] [Table 1]

[0197] [Table 2] As a result, although cell viability varies depending on the characteristics of the cancer cells, it was confirmed that L. Plantarum GB104 culture supernatant reduced cell viability in all treated human cancer cell lines compared to the untreated control (MRS).

[0198] 1.3.Confirmation of cancer cell cycle by treatment with GB104 culture supernatant

[0199] In this experiment, the culture supernatant of L. Plantarum GB104 was treated with a human colon cancer cell line, and changes in the cancer cell cycle were confirmed using a flow cytometer.

[0200] 5x10 human colon cancer cell line HCT116 cells were placed in a 6-well plate. 4 After 24 hours of incubation, the cells were aliquoted into each well and treated with 10% L. plantarum GB104 culture supernatant. The cells were then incubated at 37°C and 5% CO2 for 48 hours. The culture supernatant was prepared by culturing L. plantarum in MRS medium, centrifuging the culture to precipitate the cells, and filtering the supernatant through a 0.22 μm filter. After 48 hours, the cells were detached using trypsin-EDTA, then centrifuged and collected. The cells were fixed with 70% ethanol and stored at 4°C for at least 1 hour. After RNase A treatment and staining with propidium iodine (PI), the extent of cancer cell death was assessed by DNA content analysis using a flow cytometer (BD FACSymphony A3 Cell Analyzer). The results are shown in Figure 2.

[0201] FIG. 2 is a graph showing the results of examining the cell cycle after treating the human colon cancer cell line HCT116 with the culture supernatants of GB104 and a control strain (WCFS1).

[0202] As shown in Figure 2, the sub-G1 region of cells treated with L. plantarum GB104 culture supernatant increased by approximately 9.8-fold compared to control cells treated with MRS, confirming that GB104 induces cancer cell death.

[0203] 1.4. Early and late cell death effects of GB104 culture supernatant treatment on human cancer cell lines

[0204] In this experiment, the culture supernatant of L. Platarum GB104 was treated with a human cancer cell strain, and the cancer cell killing effect was confirmed using a flow cytometer.

[0205] 0.5-1x10 human cancer cell lines in a 6-well plate 5 Add cells to each well After 24 hours of incubation, the cells were treated with 10% L. plantarum GB104 culture supernatant and incubated at 37°C with 5% CO2 for 24 hours. The culture supernatant was prepared by culturing L. plantarum in MRS medium, centrifuging the cultures, and filtering the supernatant through a 0.22 μm filter. After 24 hours, the cells were harvested by trypsin-EDTA treatment and centrifuging. The FITC Annexin V Apoptosis Detection Kit with 7-AAD (BioLegend) was used to mix the cells with 100 μL of binding solution. Then, 5 μL of FITC Annexin V and 5 μL of 7-AAD were added and incubated at room temperature for 15 minutes in the dark. Fluorescence measurements to confirm cancer cell death were performed using a flow cytometer (BD FACSymphony A3 Cell Analyzer), and the results are shown in Figure 3. Normally viable cells are not labeled by Annexin V or 7-AAD, whereas cells in the early stage of cell death are labeled by Annexin V but not by 7-AAD. Cells in the later stage of cell death can be distinguished by being labeled by both Annexin V and 7-AAD.

[0206] FIG. 3 is a graph showing the cancer cell death effect after treating the human colon cancer cell line HCT116 or the human breast cancer cell line MDA-MB-231 with the culture supernatant of GB104 or a control strain (WCFS1).

[0207] As shown in Figure 3, L. plantarum GB104 culture supernatant significantly increased early and late cell death in treated colon cancer and breast cancer cell lines compared with the untreated control group (MRS).

[0208] This confirmed the fact that GB104 is involved in inducing cancer cell death.

[0209] Experimental Example 2. Tumor formation suppression effect by treatment with GB104 culture supernatant

[0210] In this embodiment, the tumor formation suppression effect of treatment with L. plantarum GB104 strain was confirmed in various mouse cancer cell strains.

[0211] The culture supernatant was prepared by culturing L. Platarum strains in MRS medium, precipitating the strains by centrifugation, and then collecting the supernatant. The supernatant was then filtered through a 0.22 μm filter. Specifically, 0.5-2 × 10 cells of mouse colon cancer cell strain MC-38, mouse lung cancer cell strain LLC1, and mouse breast cancer cell strain 4T1 were placed in a 6-well plate. 3 The cells were diluted with 2 mL of medium, dispensed into each well, and allowed to stabilize for 24 hours. After 24 hours, the medium was removed and replaced with fresh medium, which was treated with 1% L. plantarum GB104 culture supernatant. Fresh medium containing the culture supernatant was replaced every 2–3 days, and the cells were cultured for 7 days. The culture plates were 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. The results of colony formation were confirmed and are shown in Figure 4.

[0212] FIG. 4 is a graph showing the tumorigenesis inhibitory effect of treating the mouse MC-38 colon cancer cell line, LLC1 lung cancer cell line, and 4T1 breast cancer cell line with the GB104 culture supernatant.

[0213] As shown in Figure 4, although the tumorigenesis inhibitory ability differs depending on the characteristics of the cancer cells, L. Platarum GB104 culture supernatant was confirmed to suppress tumorigenesis in all treated mouse cancer cell lines compared to the untreated control group (MRS).

[0214] Experimental Example 3: Combined administration of GB104 strain and the anticancer drug 5-fluorouracil Confirmation of tumor growth suppression effect by

[0215] In this experiment, we confirmed the tumor therapeutic effect of the combined administration of L. plantarum GB104 and 5-fluorouracil (5-FU) in a colon carcinoma MC-38 allograft model. Six-week-old c57BL / 6 mice were introduced into the laboratory for a one-week decongestant period, and the right flank area was depilated before the experiment began at 7 weeks of age. 2 x 10 MC-38 colon cancer cell lines were inoculated into the right flank of each mouse. 5 Tumor models were established by subcutaneous injection of 100 μL of cells. Tumor size was measured using a digital caliper, and tumor volume (mm 3 )=(width 2 On the fifth day after tumor cell injection, only mice with tumor sizes within a certain range were selected and classified so that the mean tumor size of each group was equal. L. Platarum GB104 strain was then injected into the animal model at a dose of 1 × 10 per mouse. 9 CFU were orally administered daily from day 6 until just before the end of the study. The anticancer drug 5-FU (10 mg / kg) was intraperitoneally administered once a week either alone or in combination with GB104.

[0216] FIG. 5 is a graph showing the results of analyzing the synergistic effect by measuring tumor volume after combined administration of GB104 and 5-FU in an allografted MC-38 mouse colon carcinoma model.

[0217] FIG. 6 is a graph showing the results of comparative analysis of tumor volume measured after single administration and combined administration of GB104 and 5-FU in an allografted MC-38 mouse colon carcinoma model.

[0218] As shown in Figures 5 and 6, the tumor growth rate was significantly suppressed in the group administered with L. Plantarum GB104 compared to the negative control group. Furthermore, a significant reduction in tumor size due to a synergistic effect was observed in the group administered with the combination of L. Plantarum GB104 and 5-FU compared to the groups administered with either L. Plantarum GB104 or 5-FU alone.

[0219] Experimental Example 4: Confirmation of tumor growth suppression effect by combined administration of GB104 strain and immunosuppressant anti-PD-1

[0220] In this experiment, we investigated the tumor therapeutic effect of the combined administration of L. plantarum GB104 and anti-PD-1 in a colon carcinoma MC-38 allograft model. Six-week-old c57BL / 6 mice were introduced into the laboratory for a one-week decontamination period, and the right flank area was depilated before the experiment began at 7 weeks of age. The right flank of each mouse was inoculated with 2 x 10 MC-38 c57BL / 6-derived colon cancer cells. 5 Tumor models were established by subcutaneous injection of 100 μL of cells. Tumor size was measured using a digital caliper, and tumor volume (mm 3 )=(width 2 The tumor size was calculated by (x length) / 2. On the fifth day after tumor cell injection, the tumor size was 10-30 mm. 3 Only mice falling within the range were selected and randomly assigned to each group, and the L. Platarum GB104 strain was then added to the animal model at 1 × 10 per mouse. 9 CFU were orally administered daily from day 5 until the end of the study. The anti-PD-1 immunotherapy drug (5 mg / kg) was administered intraperitoneally twice a week either alone or in combination with GB104.

[0221] FIG. 7 is a graph showing the results of analyzing the synergistic effect by measuring tumor volume after co-administration of GB104 and anti-PD-1 in an allografted MC-38 mouse colon carcinoma model.

[0222] Figure 8 is a graph showing the results of measuring and comparing tumor volume after single and combined administration of GB104 and anti-PD-1 in an allografted MC-38 mouse colon carcinoma model. is.

[0223] As shown in Figures 7 and 8, compared to the negative control group (Control), the group administered L. Plantarum GB104 and anti-PD-1 in combination demonstrated a significant reduction in tumor size due to a synergistic effect compared to the group administered L. Plantarum GB104 or anti-PD-1 alone.

[0224] Experimental Example 5: Confirmation of tumor growth suppression effect by combined administration of GB104 strain and targeted anticancer drug Bevacizumab

[0225] In this experiment, the tumor therapeutic effect of combined administration of L. Plantarum GB104 strain and Bevacizumab (anti-VEGF) was confirmed in a xenograft model using immunodeficient mice. Six-week-old NOG mice were introduced into the laboratory for a one-week cleansing period, and the right flank area was depilated before the experiment began at 7 weeks of age. The right flank of each mouse was inoculated with 3 x 10 human colon cancer cell lines, HT-29. 6 Tumor models were established by subcutaneous injection of 100 μL of cells. Tumor size was measured using a digital caliper, and tumor volume (mm 3 )=(width 2 On the third day after tumor cell injection, mice with tumor sizes falling within a certain range were selected and categorized so that the mean tumor size of each group was equal. L. Platarum GB104 strain was then injected into the animal model at a rate of 1 × 10 per mouse. 9 CFU were orally administered daily from day 4 until just before the end of the study. The targeted anticancer drug Bevacizumab (10 mg / kg) was intraperitoneally administered once a week either alone or in combination with GB104.

[0226] FIG. 9 is a graph showing the results of analyzing synergistic effects by measuring tumor volume after combined treatment with GB104 and bevacisumab in a xenografted HT-29 mouse colon carcinoma model.

[0227] FIG. 10 is a graph showing the results of a comparative analysis of tumor volume measured after single administration and combined administration of GB104 and Bevacizumab in a xenografted HT-29 mouse colon carcinoma model.

[0228] As shown in Figures 9 and 10, the tumor growth rate was suppressed in the group administered with L. Plantarum GB104 compared to the negative control group. Furthermore, a significant reduction in tumor size due to a synergistic effect was confirmed in the group administered with L. Plantarum GB104 and Bevacizumab compared to the groups administered with L. Plantarum GB104 or Bevacizumab alone.

[0229] Experimental Example 6: Confirmation of tumor growth suppression effect by combined administration of GB104 strain and targeted anticancer drug Cetuximab

[0230] In this experiment, the tumor therapeutic effect of combined administration of L. plantarum GB104 strain and cetuximab (anti-EGFR) was confirmed in a xenograft model using immunodeficient mice. Six-week-old NSG mice were introduced into the laboratory for a one-week cleansing period, and the right flank area was depilated before the experiment began at 7 weeks of age. The human colon cancer cell line HT-29 was inoculated into the right flank of each mouse at a concentration of 3 x 10 per mouse. 6 Tumor models were established by subcutaneous injection of 100 μL of cells. Tumor size was measured using a digital caliper, and tumor volume (mm 3 )=(width 2 On the third day after tumor cell injection, mice with tumor sizes falling within a certain range were selected and categorized so that the mean tumor size of each group was equal. L. Platarum GB104 strain was then injected into the animal model at a rate of 1 × 10 per mouse. 9 4 days for CFU The targeted anticancer drug cetuximab (40 mg / kg) was administered intraperitoneally twice weekly either alone or in combination with GB104.

[0231] FIG. 11 is a graph showing the results of analyzing synergistic effects by measuring tumor volume after combined treatment with GB104 and Cetuximab in a xenografted HT-29 mouse colon carcinoma model.

[0232] FIG. 12 is a graph showing the results of a comparative analysis of tumor volume measured after single administration and combined administration of GB104 and Cetuximab in a xenografted HT-29 mouse colon carcinoma model.

[0233] As shown in Figures 11 and 12, a significant tumor growth inhibitory effect was confirmed in the group administered with L. Plantarum GB104 compared to the negative control group. Furthermore, a synergistic effect of tumor size reduction was confirmed in the group administered with L. Plantarum GB104 and Cetuximab in combination compared to the group administered with Cetuximab alone.

[0234] Experimental Example 7: Confirmation of tumor growth suppression effect by combined administration of GB104 strain and targeted anticancer drug Regorafenib

[0235] In this experiment, we investigated the tumor therapeutic effect of combined administration of L. Plantarum GB104 and Regorafenib in a colon carcinoma MC-38 allograft model. Five-week-old c57BL / 6 mice were introduced into the laboratory for a one-week decontamination period, and the right flank area was depilated before the experiment began at 6 weeks of age. The right flank of each mouse was inoculated with 2 x 10 MC-38 colon cancer cells. 5 Tumor models were established by subcutaneous injection of 100 μL of cells. Tumor size was measured using a digital caliper, and tumor volume (mm 3 )=(width 2 On the fifth day after tumor cell injection, only mice with tumor sizes within a certain range were selected and classified so that the mean tumor size of each group was equal. L. Platarum GB104 strain was then injected into the animal model at a dose of 1 × 10 per mouse. 9The targeted anticancer drug Regorafenib (3 mg / kg) was orally administered alone or in combination with GB104 every other day from day 5 until just before the end of the study.

[0236] FIG. 13 is a graph showing the results of analyzing the synergistic effect by measuring tumor volume and size after combined administration of GB104 and Regorafenib in an allografted MC-38 mouse colon carcinoma model.

[0237] FIG. 14 is a graph showing the results of a comparative analysis of tumor volume measured after single administration and combined administration of GB104 and Regorafenib in an allografted MC-38 mouse colon carcinoma model.

[0238] As shown in Figures 13 and 14, the tumor growth rate was significantly suppressed in the group administered with L. Plantarum GB104 compared to the negative control group. Furthermore, a significant tumor size suppression effect due to a synergistic effect was confirmed in the group administered with L. Plantarum GB104 and Regorafenib compared to the groups administered with L. Plantarum GB104 or Regorafenib alone.

[0239] Experimental Example 8: Confirmation of tumor growth suppression effect by combined administration of GB104 strain and immuno-anticancer agent GI-101

[0240] In this experiment, the tumor therapeutic effect of combined administration of L. plantarum GB104 and GI-101 was confirmed in a colon carcinoma MC-38 allograft model. Five-week-old c57BL / 6 mice were introduced into the laboratory for a one-week decontamination period, and the right flank area was depilated before the experiment began at 6 weeks of age. 2 x 10 MC-38 colon cancer cell lines were inoculated into the right flank of each mouse. 5 Tumor models were established by subcutaneous injection of 100 μL of cells. The tumor size was measured using a digital caliper. volume(mm 3 )=(width 2The tumor size was calculated by (x length) / 2. On the fifth day after tumor cell injection, the tumor size was 10-30 mm. 3 Only mice falling within the range were selected and randomly assigned to each group, and the L. Platarum GB104 strain was then added to the animal model at 1 × 10 per mouse. 9 CFU were orally administered daily from day 5 until just before the end of the study. The immunosuppressant GI-101 was subcutaneously injected at 3 mg / kg once a week from day 5 onwards either alone or in combination with GB104.

[0241] The immunoanticancer agent GI-101 was prepared based on the disclosure of Korean Patent Publication No. 10-2020-0032009. Specifically, to produce a fusion protein containing a human CD80 fragment, an Fc domain, and an IL-2 variant, a polynucleotide containing a nucleotide sequence (SEQ ID NO: 17) encoding a fusion protein containing, in order 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) with two amino acid substitutions (R38A, F42A) (SEQ ID NO: 5) was loaded into a pCGS3 vector (Sigma-Aldrich®) using the BioXP™ 3250 SYSTEM. The vector was then transfected into CHO cells (Expi-CHO®, Thermo Fisher Scientific) to express the fusion protein of SEQ ID NO: 12. After the vector was introduced, the cells were cultured for 7 days at 37°C, 125 RPM, and 8% CO2, and then the culture medium was collected and the fusion protein was purified.

[0242] FIG. 15 is a graph showing the results of analyzing synergistic effects by measuring tumor volume and weight after combined treatment with GB104 and GI-101 in an allografted MC-38 mouse colon carcinoma model.

[0243] FIG. 16 is a graph showing the results of comparative analysis of tumor volume measured after single administration and combined administration of GB104 and GI-101 in an allografted MC-38 mouse colon carcinoma model.

[0244] As shown in Figures 15 and 16, compared to the negative control group (Control), the group administered with a combination of L. Plantarum GB104 and GI-101 showed a significant reduction in tumor size, demonstrating enhanced antitumor efficacy compared to the group administered with L. Plantarum GB104 or GI-101 alone.

[0245] Experimental Example 9: Confirmation of tumor growth suppression effect by combined administration of GB104 strain and immuno-anticancer agent GI-102

[0246] In this experiment, the tumor therapeutic effect of the combined administration of L. plantarum GB104 and GI-102 was confirmed in a colon carcinoma MC-38 allograft model. Five-week-old c57BL / 6 mice were introduced into the laboratory for a one-week decontamination period, and the right flank area was depilated before the experiment began at 6 weeks of age. The right flank of each mouse was inoculated with 2 x 10 MC-38 colon cancer cell lines. 5 Tumor models were established by subcutaneous injection of 100 μL of cells. The tumor size was measured using a digital caliper. volume(mm 3 )=(width 2 Calculate the size by (xlength) / 2 On the fifth day after tumor cell injection, the tumor size was 10-30 mm. 3 Only mice falling within the range were selected and randomly assigned to each group, and the L. Platarum GB104 strain was then added to the animal model at 1 × 10 per mouse. 9 CFU were orally administered daily from day 5 until just before the end of the study. The immunosuppressant GI-102 was subcutaneously injected at 3 mg / kg on days 5 and 15 either alone or in combination with GB104.

[0247] Here, the immunoanticancer agent GI-102 was prepared with reference to the disclosure of Korean Patent Publication No. 10-2020-0032009. Specifically, to produce a fusion protein containing a human CD80 fragment, an Fc domain, and an IL-2 mutant with three amino acid substitutions, a polynucleotide containing, in order 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 mutant with three amino acid substitutions (3M) (R38A, F42A, E61R) (SEQ ID NO: 7) in this order from the N-terminus (SEQ ID NO: 18) was transfected with BioXP®. The vector was loaded into a pCGS3 vector (Sigma-Aldrich®) using the 3250 SYSTEM. The vector was then introduced into CHO cells (Expi-CHO®, Thermo Fisher Scientific) to express the fusion protein of SEQ ID NO: 14. After vector introduction, the cells were cultured at 37°C, 125 RPM, and 8% CO for 7 days, after which the culture medium was collected and the fusion protein was purified.

[0248] FIG. 17 is a graph showing the results of analyzing synergistic effects by measuring tumor volume and weight after combined treatment with GB104 and GI-102 in an allografted MC-38 mouse colon carcinoma model.

[0249] FIG. 18 is a graph showing the results of comparative analysis of tumor volume measured after single administration and combined administration of GB104 and GI-102 in an allografted MC-38 mouse colon carcinoma model.

[0250] As shown in Figures 17 and 18, tumor size was significantly reduced in the group administered with the combination of L. Plantarum GB104 and GI-102 compared to the negative control group. Furthermore, the group administered with the combination of L. Plantarum GB104 and GI-102 showed a greater effect in reducing tumor size than the groups administered with either L. Plantarum GB104 or GI-102 alone. This confirmed that the combined administration of L. Plantarum GB104 and GI-102 enhanced the antitumor efficacy.

[0251] Experimental Example 10. Tumor growth suppression effect of triple combined administration of GB104 strain, immunological anticancer drug GI-101, and chemical anticancer drug 5-FU

[0252] In this example, the tumor therapeutic effect of triple combined administration of the L. plantarum GB104 strain, the immunological anticancer agent GI-101, and the chemical anticancer agent (5-Fluorouracil, 5-FU) was confirmed in a colon carcinoma MC-38 allograft model. Six-week-old c57BL / 6 mice were introduced into the laboratory for a one-week cleansing period, and the right flank area was depilated before the experiment began at 7 weeks of age. The c57BL / 6-derived colon cancer cell strain MC-38 was inoculated into the right flank of each mouse at 2 x 10 5 Tumor models were established by subcutaneous injection of 100 μL of cells. Tumor size was measured using a digital caliper, and tumor volume (mm 3 )=(width 2 The tumor size was calculated by (x length) / 2. On the 6th day after tumor cell injection, the tumor size was 10-30 mm. 3 Only mice falling within the range were selected and randomly assigned to each group, and the L. Platarum GB104 strain was then added to the animal model at 1 × 10 per mouse. 9 CFU were orally administered daily from day 6 until just before the end of the study. The immunosuppressant GI-101 was administered subcutaneously at 3 mg / kg once a week from day 6, for a total of three doses. The chemo-anticancer agent 5-FU was administered intraperitoneally at 10 mg / kg once a week from day 6, for a total of three doses.

[0253] FIG. 19 is a graph showing the results of analyzing synergistic effects by measuring tumor volume after triple combination treatment of GB104, GI-101, and 5-FU in an allografted MC-38 mouse colon carcinoma model.

[0254] FIG. 20 is a graph showing the results of a comparative analysis of tumor volume measured after single administration and combined administration of GB104, GI-101, and 5-FU in an allografted MC-38 mouse colon carcinoma model.

[0255] As shown in Figures 19 and 20, on day 26 after tumor inoculation, the GB104 monotherapy group showed a 29.72% tumor growth inhibition compared to the negative control group (PBS), the GB104 + GI-101 dual combination therapy group showed a 50.01% tumor growth inhibition, and the GB104 + GI-101 + 5-FU triple combination therapy group showed a 63.92% superior antitumor effect.

[0256] This confirmed that in the treatment of colon cancer, the antitumor efficacy was strengthened through significant suppression of tumor growth in the group administered GB104 strain alone as well as in the group administered in combination with the immunological anticancer agent GI-101 and the existing chemical anticancer agent 5-FU. [Accession number]

[0257] Name of depository institution: Korea Institute of Bioscience and Biotechnology Accession number: KCTC14107BP Date of acceptance: 20200114

Claims

1. The present invention relates to a method for treating a cancer, comprising: a first active substance comprising a Lactobacillus plantarum strain, a culture of the strain, a disrupted product of the strain, or a mixture thereof; and a second active substance comprising an immunological anti-cancer agent as a first anti-cancer agent, as active ingredients; or A pharmaceutical composition for preventing or treating cancer, comprising the first active substance as an active ingredient, and the second active substance administered in combination.

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

3. The pharmaceutical composition for preventing or treating cancer according to claim 1 , wherein the first active substance induces cell death 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 the strain deposited under accession number KCTC14107BP.

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

7. 2. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein the immunoanticancer agent is a fusion protein comprising a CD80 protein or a fragment thereof and an IL-2 protein or a variant thereof.

8. 8. The pharmaceutical composition for preventing or treating cancer according to claim 7, wherein the IL-2 mutant has at least one amino acid substitution at positions 38, 42, 45, 61, and 72 in 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 mutant has the amino acid sequence of SEQ ID NO: 5 to 8.

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

3.

11. 8. The pharmaceutical composition for preventing or treating cancer according to claim 7, wherein the fusion protein has the following structural formula (I) or (II): N'-X-[linker 1]n-Fc domain-[linker 2]m-Y-C'(I) N'-Y-[Linker 1]n-Fc domain-[Linker 2]m-X-C' (II) In this case, in the structural formulas (I) and (II), N' is the N-terminus of the fusion protein; the C' is the C-terminus of the fusion protein; X is a CD80 protein or a fragment thereof; Y is an IL-2 protein or a variant thereof; said linker 1 and linker 2 are peptide linkers; The n and m each independently represent 0 or 1.

12. 8. The pharmaceutical composition for preventing or treating cancer according to claim 7, wherein the fusion protein has 90% or more sequence identity with the amino acid sequences of SEQ ID NOs: 12 to 15.

13. The pharmaceutical composition for preventing or treating cancer according to claim 7, wherein the fusion proteins are formed by binding two identical proteins to form a homodimer.

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

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

16. 2. 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. 2. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein the first and second active substances are administered to an individual at intervals of 1 to 10 days.

18. 2. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein the first active substance is administered to an individual at intervals of 1 to 3 days, and the second active substance is administered to an 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 group consisting of a chemical anticancer agent, a targeted anticancer agent, and an immune anticancer agent.

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

22. 21. The pharmaceutical composition for the prevention or treatment of cancer according to claim 20, wherein the anticancer chemotherapy agent is any one selected from the group consisting of mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, thiotepa, altretamine, procarbazine, busulfan, streptozocin, carmustine, lomustine, dacarbazine, cisplatin, carboplatin, oxaliplatin, docetaxel, velban, Oncovin, navelbine, 5-fluorouracil, capecitabine, cytarabine, gemcitabine, fludarabine, methotrexate, pemetrexed, mercaptopurine, hycamtin, irinotecan, bepside, paclitaxel, bleoscein, adriamycin, etoposide and cervidin.

23. The method of claim 20, wherein the targeted anticancer drug targets any protein selected from the group consisting of epidermal growth factor receptor (EGFR), vascular endothelial growth factor receptor (VEGFR), CD20, CD38, RNAK-L, BTK, Bcr-abl, PDGFR / FGFR family, MEK / RAF, HER2 / Neu, ubiquitin, JAK, MAP2K, ALK, PARP, transforming 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. Prophylactic or therapeutic pharmaceutical compositions.

24. The targeted anticancer drug is selected from the group consisting of olaratumab, erlotinib, panitumumab, trastuzumab, trastuzumab emtansine, pertuzumab, cetuximab, rituximab, bevacizumab, axitinib, lenvatinib, ramucirumab, aflibercept, obinutuzumab, daratumumab, denosumab, ibrutinib, dasatinib, and radocinib. nilotinib, imatinib, bosutinib, galunisertib, bactosertib, nintedanib, sunitinib, sorafenib, cabozantinib, regorafenib, masitinib, semaxanib, ceritinib, tivozanib, brigatinib, vandetanib, pazopanib, trametinib, temsirolimus, dabrafenib, dacomitinib, and 21. The pharmaceutical composition for the prevention or treatment of cancer according to claim 20, wherein the pharmaceutical composition is any one selected from the group consisting of patinib, lapatinib, neratinib, lenalidomide, ocimatinib, ixazomib, olmutinib, everolimus, ruxolitinib, lestaurtinib, pacritinib, cobimetinib, selumetinib, binimetinib, bortezomib, alectinib, crizotinib, venetoclax, bensentinib, gilteritinib, selpercatinib, pralsetinib, vemurafenib, olaparib, talazoparib, niraparib, rucaparib, azacitidine, decitabine, guadecitabine, gefitinib, abemaciclib, ribociclib, palbociclib, and DMXAA.

25. 21. The pharmaceutical composition for preventing or treating cancer according to claim 20, wherein the anti-cancer immunoagent is an antibody against any one selected from the group consisting of 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 immunological anticancer agent is atezolizumab, avelumab, davalumab, nivolumab, pembrolizumab, abagovomab, adecatumumab, afutuzumab, alemtuzumab, anatumomab mafenatox, mepolizumab, blinatumomab, BMS-936559, catumaxomab, cemiplimab, epacadostat, epratuzumab, indoximod, inotuzumab, inotuzumab ozoga The pharmaceutical composition for preventing or treating cancer according to claim 20, wherein the compound is any one selected from the group consisting of mycin, intelumumab, ipirlimumab, isatuximab, lambrolizumab, MED14736, MPDL3280A, obinutuzumab, ocaratulumab, ofatumumab, olaratumab, fidelizumab, rituximab, ticilimumab, samarijuana, 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. 20. 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. 2. The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein the cancer is any one selected from the group consisting of gastric cancer, liver cancer, lung cancer, colon 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.

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

31. The present invention relates to a method for treating a cancer, comprising: a first active substance comprising a Lactobacillus plantarum strain, a culture of the strain, a disrupted product of the strain, or a mixture thereof; and a second active substance comprising an immunological anti-cancer agent as a first anti-cancer agent, as active ingredients; or A health functional food for preventing or ameliorating cancer, comprising the first active substance as an active ingredient and administered in combination with the second active substance.

32. The health functional food for preventing or ameliorating cancer according to claim 31, wherein the strain is the strain deposited under accession number KCTC14107BP.

33. The health functional food for preventing or ameliorating cancer according to claim 31, wherein the immunoanticancer agent is a fusion protein comprising an IL-2 protein or a mutant thereof and a CD80 protein or a fragment thereof.

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

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

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

39. 32. The health functional food for preventing or ameliorating cancer according to claim 31, wherein the cancer is any one selected from the group consisting of stomach cancer, liver cancer, lung cancer, colon 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.

40. The health functional food for preventing or ameliorating cancer according to claim 39, wherein the colon cancer occurs in any site selected from the group consisting of the ascending colon, transverse colon, descending colon, sigmoid colon, and rectal mucosa.

41. A method for preventing or treating cancer, comprising the step of administering to an individual in need thereof a composition comprising, as active ingredients, a first active substance comprising an effective amount of a Lactobacillus plantarum strain, a culture of said strain, a disrupted product of said strain, or a mixture thereof, and a second active substance comprising a first anti-cancer agent, which is an immunological anti-cancer agent, or a composition comprising said first active substance as an active ingredient and said second active substance administered in combination.

42. Lactobacillus planta for the manufacture of pharmaceutical preparations for preventing or treating cancer Use of a composition comprising as active ingredients a first active substance comprising a Lactobacillus plantarum strain, a culture of said strain, a disrupted product of said strain, or a mixture thereof, and a second active substance comprising an immunological anticancer agent that is a first anticancer agent, or a composition comprising said first active substance as an active ingredient and said second active substance administered in combination.

43. Use of a composition comprising, as active ingredients, a first active substance comprising a Lactobacillus plantarum strain, a culture of said strain, a disrupted product of said strain, or a mixture thereof, and a second active substance comprising an immunological anticancer agent that is a first anticancer agent, or a composition comprising said first active substance as an active ingredient and said second active substance administered in combination, for the manufacture of a health functional food for preventing or treating cancer.

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