Composition for preventing or treating cancer using combination therapy containing a Lactobacillus plantarum strain and an anticancer drug

Abstract

The present invention relates to a pharmaceutical composition for cancer treatment in which a Lactobacillus plantarum strain is administered in combination with an anticancer drug. The Lactobacillus plantarum strain or a culture medium of said strain can not only suppress the proliferation of cancer cells or induce the death of cancer cells, but also exhibits a synergistic effect in suppressing the proliferation of cancer cells and tumors when administered in combination with an anticancer drug, and can be usefully used in the form of a pharmaceutical composition or a health functional food for the prevention, treatment or improvement 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 anticancer agent. [Background technology]

[0002] Microbiome is a compound word of microbe and genome, and human microbiome means the genome of all microorganisms living in the human body. The human body is composed of various types of microorganisms in several parts, but 70% of the human microbiome is distributed in the digestive tract, with the largest number of microorganisms living in the large intestine. The microbiome is closely related to the human immune system and physical development, and it has been steadily reported that it is highly related to various diseases such as metabolic diseases such as obesity and diabetes, inflammatory bowel disease, and depression. After the Human Genome Project was initially completed in 2002, attempts were made to confirm the relationship between genes and intractable diseases, and it was confirmed that environmental factors, especially the microbiome environment, may have a large influence on the disease. Microbiome therapeutics grow in the intestines as living organisms and affect the human body through interactions with the cells of the human body. For this reason, they are attracting much attention as an innovative new drug that can improve the low efficacy and high recurrence rate of intractable diseases against existing drugs. In addition, in December 2022, the US FDA approved Ferring Pharmaceuticals' REBYOTA (registered trademark), the first microbiome-based treatment for Clostridium difficile infection (CDI), marking the beginning of the practical application of microbiome therapeutics, and Seres Therapeutics' SER-109 is also under FDA review as the first oral microbiome therapeutic.

[0003] Globally, cancer is considered one of the major health issues. It is the second leading cause of death worldwide and, if it occurs, requires intensive treatment and steady management, making it a disease that places a very high burden on individuals and socio-economically. As of 2021, the global anti-cancer drug market is worth approximately KRW 225 trillion, and is expected to grow at an average annual rate of 11% in the future, reaching approximately KRW 470 trillion in 2028, due to the increase in the number of patients, the growth of immune anti-cancer drugs, and the emergence of new treatments.

[0004] Conventional cancer treatment involves a combination of chemotherapy, surgery, hormonal therapy and / or radiation therapy to remove the patient's neoplastic cells. Cancer chemotherapy is preferably based on the use of drugs that kill replicating cells faster than the formulation kills the patient's normal cells. Anticancer drugs show cytotoxicity to cells by intervening in the metabolic pathways of cancer cells and directly acting on DNA to block the DNA replication, transcription and translation processes or to interfere with the synthesis of nucleic acid precursors, thereby inhibiting cell division. Therefore, these anticancer drugs do not only selectively act on cancer cells, but also cause fatal damage to normal cells, especially tissue cells with active cell division, causing various serious side effects such as vomiting, bone marrow depression, gastrointestinal disorders, alopecia, diarrhea, liver and kidney toxicity. In response to this, immune anticancer drugs (or immune checkpoint inhibitors) have been greatly highlighted as third-generation anticancer drugs, but the requirements for their use are very restrictive. Therapeutic effects are generally seen in less than 30% of cases. Therefore, in order to completely conquer the disease of cancer, more attention needs to be paid to research on the mechanism of immune anticancer drug efficacy enhancement by intestinal microorganisms.

[0005] Recently, combination therapy has become popular to enhance the efficacy of treatment for diseases. Combination therapy is the use of two or more drugs or methods simultaneously or in relatively rapid succession. The side effects of two or more treatment methods used in combination therapy may be additive or less than additive, but the therapeutic effects may be additive or greater than additive. The effect of each component administered in combination is expected to be greater than the sum of the effects of each component administered alone. Therefore, there is a need to develop combination therapy that can synergistically increase the effect of treatment using a single component. 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, 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 anti-cancer drug, or comprising the first active substance as an active ingredient and the second active substance being administered in combination.

[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 homogenate of said strain, or a mixture thereof, and a second active substance comprising an anticancer drug, 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, comprising 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 an anti-cancer drug, or comprising as an active ingredient the first active substance and the second active substance administered in combination.

[0009] Another aspect is to provide a method for delivering a drug into an individual comprising administering to an individual in need thereof a composition comprising 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 an anti-cancer drug, or the first active substance as an active ingredient is co-administered with the second active substance.

[0010] Another aspect is to provide 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 a Lactobacillus plantarum strain, a culture of said strain, a homogenate of said strain, or a mixture thereof, and a second active substance comprising an anti-cancer drug, or the first active substance as an active ingredient in combination with the second active substance.

[0011] Another aspect provides use of a composition comprising 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 an anti-cancer drug, 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 pharmaceutical preparation for preventing or treating cancer.

[0012] Another aspect relates to a method for producing a health functional food for preventing or improving cancer. The present invention provides use of a composition comprising 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 an anticancer drug, or a composition comprising the first active substance as an active ingredient and the second active substance administered in combination. [Means for solving the problem]

[0013] One aspect provides a Lactobacillus strain, specifically Lactobacillus plantarum strain GB104, having anti-cancer activity, for example anti-cancer activity against colon cancer.

[0014] Lactobacillus is a gram-positive aerobic or facultative anaerobic bacillus microorganism that is widely distributed in nature. Microorganisms belonging to the Lactobacillus genus include Lactobacillus plantarum and Lactobacillus sakae. The present inventors conducted research to develop a new strain with excellent anti-cancer effects and selected Lactobacillus plantarum GB104 as an anti-cancer candidate strain. The strain was deposited at the Korea Institute of Bioscience and Biotechnology Biological Resources Center on January 14, 2020 under the deposit number KCTC14107BP. The strain corresponds to a probiotic strain and is harmless to the human body and can be used without side effects.

[0015] The Lactobacillus species has been renamed Limosilactobacillus or Lactiplantibacillus, and the modified strain names herein may be used interchangeably, for example, Lactobacillus plantarum has been renamed Lactiplantibacillus plantarum.

[0016] As used herein, the term "Lactobacillus plantarum" "L. plantarum GB104" may be used in conjunction with L. plantarum GB104 strain or Lactobacillus plantarum GB104 strain (accession number: KCTC14107BP).

[0017] In one embodiment, the strain may be the strain deposited under accession number KCTC14107BP.

[0018] In one embodiment, the strain may be a strain comprising a 16S rRNA gene consisting of the nucleotide sequence of SEQ ID NO:1.

[0019] 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.

[0020] In one embodiment, the strain may be a mutant strain of a naturally occurring strain.

[0021] In one embodiment, the strain may be a live cell, a killed cell or a cytoplasmic fraction obtained by disrupting the strain, preferably a live cell.

[0022] The term "culture" as used herein may be used interchangeably with "culture supernatant," "culture supernatant," "conditioned culture," or "conditioned medium," and refers to the culture medium in which a Lactobacillus strain is It may mean the whole medium containing the strain, its metabolites, extra nutrients, etc., obtained by culturing the strain for a certain period of time in a medium capable of supplying nutrients so that the strain can grow and survive in vitro. The culture means 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 may 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.

[0023] The term "lysate" as used herein may be used interchangeably with "dissolved product" to refer to a solution or suspension in an aqueous medium of cells of a microorganism, such as Lactobacillus plantarum, that have been broken down. A cell lysate contains macromolecules, such as DNA, RNA, proteins, peptides, carbohydrates, lipids, etc., and / or micromolecules, such as amino acids, sugars, fatty acids, etc., or fractions thereof. In addition, the lysate contains cell debris, which may be of smooth or granular structure.

[0024] 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.

[0025] 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.

[0026] 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 homogenate of the strain, or an extract of the strain, the culture, and the homogenate may have anticancer activity.

[0027] Specifically, the anti-cancer activity may be an activity that delays the onset of tumors or inhibits the rate of tumor growth.

[0028] In one embodiment, the first active agent may have tumor growth inhibitory activity.

[0029] In one embodiment, the first active agent may induce cell death in cancer cells.

[0030] According to one embodiment, the anti-cancer activity may be a cancer cell growth inhibitory activity in which the survival rate of the cancer cells is reduced by 90% to 98% when the supernatant of Lactobacillus plantarum GB104 strain is treated in colon cancer cells HCT116, 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.

[0031] According to one embodiment, the anticancer activity is an activity of inducing cell death in cancer cells, and when the supernatant of Lactobacillus plantarum GB104 strain is treated with colon cancer cells HCT116, 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.

[0032] In one embodiment, the first active agent may have anti-cancer activity, specifically anti-cancer activity against colon cancer.

[0033] The term "cancer" as used herein refers to a physiological condition in an animal that is typically characterized by abnormal or unregulated cell growth. Cancer and cancer pathology can include, for example, For example, it may be associated with 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.

[0034] The cancer can be a gastrointestinal cancer or a non-gastrointestinal cancer.

[0035] The gastrointestinal cancer is a malignant tumor occurring in the gastrointestinal tract, such as the esophagus, stomach, small intestine, or large intestine, and 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, gastric cancer, small intestine cancer, large intestine cancer, colon cancer, anal cancer, and rectal cancer, but is not limited to these, and one example may be large intestine cancer.

[0036] The non-digestive tract cancer includes, without limitation, malignant tumors occurring in organs other than the digestive tract or digestive system, and may be, for example, but not limited to, 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.

[0037] In one embodiment of the present invention, the cancer may be colon cancer, which includes malignant tumors occurring at one or more sites selected from the group consisting of ascending colon, transverse colon, descending colon, sigmoid colon, and rectal mucosa. The colon cancer 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 agent may comprise a Lactobacillus plantarum strain as the sole active ingredient or may comprise one or more pharma- ceutically acceptable carriers, excipients or diluents.

[0039] In one embodiment, the second active agent may include an anti-cancer agent.

[0040] The anti-cancer agent may be selected from the group consisting of chemo-anti-cancer agents for chemotherapy, targeted anti-cancer agents, anti-cancer viruses, antibody therapeutic agents, immune anti-cancer agents (hereinafter, may be used interchangeably with "immune checkpoint inhibitors"), and combinations thereof. Specifically, the anti-cancer 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] The term "antibody" as used herein refers to any isotype of intact immunoglobulin, or an antigen-binding fragment that can compete with an intact antibody for binding to a target antigen. Examples include chimeric, humanized, fully human and bispecific antibodies, or antigen-binding fragments thereof. The antibody is itself a type of antigen-binding protein. The antibody or antigen-binding fragment thereof may be derived from a single source or 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 may be a hybrid antibody. Antibodies can be produced by hybridomas, recombinant techniques, or by enzymatic or chemical cleavage of intact antibodies. 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.

[0042] As used herein, a "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.

[0043] As used herein, "heavy chain" includes full-length heavy chains 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 at the amino terminus of the heavy chain polypeptide, the CH domain is at the carboxyl terminus, and CH3 is located closest to the carboxyl terminus. The heavy chain includes isotypes of IgG (including IgG1, IgG2, IgG3, and IgG4 subtypes), IgA (including IgA1 and IgA2 subtypes), and IgM and IgE.

[0044] As used herein, an "antigen binding fragment" of an antibody or immunoglobulin chain (heavy or light chain) includes a portion of an antibody that lacks some amino acids compared to the full-length chain but is capable of specifically binding to an antigen. The fragment can be considered to have biological activity in terms of its ability to specifically bind to a target antigen or to compete with other antibodies or antigen binding fragments for binding to a particular epitope. In one aspect, the fragment includes at least one CDR present in a full-length light or heavy chain, and in some embodiments, a truncated heavy and / or light chain or a portion thereof. The 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 from any mammal, including, but not limited to, human, mouse, rat, camelid, or rabbit.

[0045] 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.

[0046] As used herein, a "Fab fragment" consists of one light chain and one heavy chain that contain 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 molecules of Fab linked by a flexible linker.

[0047] As used herein, a "Fab' fragment" includes a Fab fragment plus the region between the CH1 and CH2 domains of the heavy chain, which can form a disulfide bond between the two heavy chains of the two molecules of the Fab' fragment to generate an F(ab')2 molecule.

[0048] As used herein, a "F(ab')2" fragment consists of two light chains and As mentioned above, it contains two heavy chains including the variable region, CH1 and a part of the constant region between the CH1 and CH2 domains, which includes two intrachain disulfide bonds. Thus, the F(ab')2 fragment is composed of two Fab' fragments, which are held together by disulfide bonds between them.

[0049] The term "Fv region" as used herein refers to a fragment of an antibody that contains the variable regions of the heavy and light chains but does not contain the constant region. An sdFV is a fragment of a heavy and light chain linked by a disulfide bond. An scFc is a fragment of a single stranded variable region (Fv) of a heavy and light chain linked via a flexible linker. An scFv-Fc is a fragment of an Fc linked to an scFV. The minibody is a fragment of a CH3 linked to an scFV. The diabody contains the scFVs of both molecules.

[0050] A "short-chain antibody (scAb)" herein is a single polypeptide chain that contains one variable region of a heavy or light chain constant region, with the heavy and light chain variable regions connected by a flexible linker.

[0051] A "domain antibody (dAb)" herein is an immunologically functional immunoglobulin fragment that contains only the variable region of a heavy chain or the variable region of a light chain.

[0052] A "bivalent antigen-binding protein" or "bivalent antibody" as used herein refers to a protein that contains two antigen-binding sites. The two antigen-binding sites in such a bivalent antibody can have the same antigen specificity or can be bispecific antibodies that each bind to a different antigen.

[0053] A "multi-specific antigen-binding protein" or "multi-specific antibody" herein is one that targets two or more antigens or epitopes.

[0054] As used herein, a "bispecific" or "dual-specific" antigen-binding protein or antibody is a hybrid antigen-binding protein or antibody that has two different antigen-binding sites. Such bispecific antibodies are a type of multispecific antigen-binding protein or antibody and can be produced by a variety of known methods, such as fusion of hybridomas or linkage of Fab' fragments.

[0055] In one embodiment, the additional polypeptide comprises an additional antibody or fragment thereof. Additional antibodies include, for example, intact IgG, IgE and IgM, bispecific or multispecific antibodies (such as Zybodies®), single chain Fv, polypeptide-Fc fusions, Fab, cameloid antibodies, masked antibodies (such as Probodies®), Small Modular ImmunoPharmaceuticals ("SMIPs™"), single chain or tandem diabodies (TandAb®), VHHs (including but not limited to those described in this disclosure), Anticalins®, Nanobodies®, minibodies, BiTEs®, ankyrin repeat proteins or DARPINs®, Avimers®, DARTs, TCR mimetic antibodies, These include Adnectins, Affilins, Trans-bodies, Affibodies, TrimerX, 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 immune suppressive pathway.

[0056] 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, p15, overexpressed embryonic antigens such as CEA, overexpressed oncogenes and mutated tumor suppressor genes such as p53, Ras, HER-2 / neu, unique tumor antigens derived from chromosomal translocations such as BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, and viral antigens such as Epstein-Barr virus antigen EBVA and human papillomavirus (HPV) antigens E6 and E7. Other tumor antigens include TSP-180, MAGE-4, MAGE-5, MAGE-6, RAGE, NY-ESO, erbB, p185erbB2, p180erbB-3, c-met, nm-23H1, PSA, TAG-72, 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.

[0057] 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.

[0058] The term "chemical anticancer agent" in this specification is also called an antitumor agent or a cytotoxic agent. It is a general term for drugs that exhibit anticancer activity mainly by acting directly on DNA to block the DNA replication, transcription, and translation processes, or by interfering with the synthesis of nucleic acid precursors in metabolic pathways and inhibiting cell division. The antitumor drugs act not only on tumor cells but also on normal cells, exhibiting cytotoxicity. Chemical anticancer agents can be used in maintenance therapy. The term "maintenance therapy" in this specification refers to the initial stage of anticancer treatment. This refers to a treatment method in which drugs are used to treat cancer after chemotherapy, in order to prevent or delay the recurrence of cancer.

[0059] Specifically, the chemo-cancer agent may be any one selected from the group consisting of alkylating agents, microtubule inhibitors, antimetabolites and topoisomerase inhibitors. The alkylating agent may be any one selected from the group consisting of mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, thiotepa, altretamine, procarbazine, busulfan, streptozotocin, carmustine, lomustine, dacarbazine, 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 Fluorouracil, Capecitabine, Cytarabine, Gemcitabine, Fludarabine, Methotrexate, Pemetrexed, and Mercaptopurine. The topoisomerase inhibitor can be any selected from the group consisting of Hycamtin, Camptosar, Vepesid, Paclitaxel, Blenoxane, Adriamycin, and Cerubidine.

[0060] The term "targeted anticancer drug" in this specification refers to a therapeutic agent that targets a specific protein or specific genetic alteration that is abundant only in cancer cells, blocking signals involved in the growth and development of cancer and killing cancer cells specifically. They are classified into monoclonal antibodies that react outside the cell and small molecule substances that act inside the cell. Monoclonal antibodies are anticancer drugs that block cancer cell-inducing signals transmitted outside the cell and act on initiation signals related to proliferation, death, etc., while small molecule substances act on complex signal transduction that occurs inside the cell.

[0061] Specifically, the target protein may be EGFR, VEGFR, CD20, CD38, RNAK-L, BTK, Bcr-abl, PDGFR / FGFR family, MEK / RAF, HER2 / Neu, ubiquitin, JAK, MAP2K, ALK, PARP, TGFβRI, proteasome, Bcl-2, C-Met, VR1, VR2, VR3, c-kit, AXL, RET, Braf, DNMT, CDK4 / 6, STING, and the like.

[0062] The targeted anticancer drug is abciximab. adalimumab, basiliximab, bezlotoxumab, canakinumab, daclizumab, denosumab, efalizumab, golimumab, inflectra, natalizumab, olaratumab, omalizumab, palivizumab, panitumumab, trastuzumab, pertuzumab, cetuximab, It may be any one selected from the group consisting of rituximab, tocilizumab, secukinumab, ustekinumab, bevacizumab, avelumab, CDNs, SB11285 and DMXAA.

[0063] The term "epidermal growth factor receptor (EGFR)" as used herein refers to a cell membrane receptor that regulates cell growth, division, survival, and death, and expression of EGFR is increased in tumor tissues in various cancers. Tumor tissues with increased EGFR are known to have high invasiveness, metastasis, and resistance to anticancer drugs. In one embodiment, the EGFR inhibitor, a substance that inhibits EGFR, may be cetuximab, trastuzumab, pertuzumab, gefitinib, erlotinib, or panitumumab.

[0064] The term "vascular endothelial growth factor receptor (VEGFR)" in this specification refers to a cell membrane receptor of angiogenic factors that induce angiogenesis, and VEGFR inhibitors inhibit the angiogenesis to suppress tumor growth and metastasis. One embodiment of the VEGF inhibitor or VEGFR inhibitor can be Axitinib, Lenvatinib, Bevacizumab, Ramucirumab or Aflibercept.

[0065] The term "CD20 (B lymphocyte antigen CD20)" is used herein as a target protein for treating B cell lymphoma as a protein expressed on the surface of B cells. The CD20 target inhibitor can be Rituximab or Obinutuzumab.

[0066] 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.

[0067] The term "RNAK-L (Receptor activator of nuclear factor kappa-B ligand)" as used herein refers to the RANK receptor expressed on the surface of osteoclasts, which acts to cause bone destruction when it binds to and is activated by a ligand. RANK-L inhibitors are primarily used for cancer patients suffering from bone metastasis and osteoporosis, and specifically may be Denosumab.

[0068] The term "BTK (Bruton's tyrosine kinase)" as used herein is an enzyme involved in the proliferation of B cells, which when overexpressed can lead to hematological cancers. One embodiment of a BTK targeting inhibitor can be Ibrutinib.

[0069] The term "Bcr-abl" in this specification is 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 the protein can be Dasatinib, Nilotinib, Imatinib or Bosutinib.

[0070] As used herein, the term "tumor growth factor β receptor" refers to The term "TGFβR" refers to a cell membrane receptor for tumor growth factors, which regulates the growth, migration, differentiation and death of epithelial and hematopoietic cells. The TGFβR targeting inhibitors include, but are not limited to, Galunisertib or Vactosertib.

[0071] The term "PDGFR (Platelet derived growth factor)" as used herein 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 FGFR genes are common, and such mutants are commonly found in breast cancer, uterine cancer, ovarian cancer, cervical cancer, and the like. The inhibitor targeting PDGFR or FGFR can be Nintedanib, Sunitinib, Sorafenib, Cabozantinib, Lenvatinib, Regorafenib, Masitinib, Semaxanib, Tivozanib, Vandetanib, Axitinib, or Pazopanib.

[0072] The term "MEK / RAF" as used herein refers to an intracellular signaling mediator involved in cell proliferation, cell cycle regulation, cell survival, angiogenesis, cell migration, etc., and is hyperactivated in cancer cells. The inhibitor targeting MEK / RAF may be trametinib or dabrafenib.

[0073] The term "HER-2 / neu (Human epidermal growth factor receptor 2)" as used herein regulates cell proliferation by activating PI3K / AkT. It is overexpressed in metastatic breast cancer, ovarian cancer, and other cancers, and is known to induce resistance to anticancer drugs. The Her2 / neu targeted anticancer drug may be Trastuzumab, Afatinib, Lapatinib, or Neratinib.

[0074] The term "ubiquitin" as used herein refers to a ubiquitin that binds to other proteins and maintains cell homeostasis by 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 anticancer activity. Specifically, inhibitors targeting ubiquitin or proteasomes may be lenalidomide or ixazomib.

[0075] The term "JAK (Janus kinase)" as used herein refers to a higher-rank protein of STAT, a transcription factor that regulates cell proliferation, cell survival, cell migration and immune response, and it is known that JAK inhibitors reduce cell proliferation and induce cell death through inhibition of STAT activity. The JAK target inhibitor may be Ruxolitinib, Lestaurtinib or Pacritinib.

[0076] As used herein, the term "MAP2K (Mitogen-activated protein kinase kinase)" refers to a kinase that phosphorylates MAPK and promotes cell proliferation, It is an intracellular signaling mediator involved in cell cycle regulation, cell survival, angiogenesis, cell migration, etc., and is overactivated in cancer cells. The MAP2K target inhibitor can be Cobimetinib, Selumetinib, Trametinib, or Binimetinib.

[0077] The term "ALK (Anaplastic lymphoma kinase)" as used herein refers to a signal transduction mediator that promotes cell proliferation, cell migration, and angiogenesis, and inhibits cell death, and is hyperactivated in various cancer tissues. The ALK target inhibitor can be Alectinib or Crizotinib.

[0078] The term "Bcl-2" as used herein refers to a protein that inhibits cell death and is overexpressed or overactivated in various cancer tissues. The inhibitor targeting Bcl-2 can be venetoclax.

[0079] The term "C-Met" as used herein refers to a receptor for hepatocyte growth factor (HGF), which activates signal transduction associated with cell growth, formation, motility, survival, angiogenesis, etc. C-Met targeted anticancer drugs can be crizotinib or cabozantinib.

[0080] In this specification, the term "VR (Vanilloid receptor)" refers to TRPV (Transient Receptor Potential Vanilloid Receptor). VRs are also known as VR-associated VR domains (VR domains VR1, VR2, VR3, VR4, VR5 and VR6) and exist in the forms VR1, VR2, VR3, VR4, VR5 and VR6. VRs are known to regulate the proliferation, death, migration, invasion and angiogenesis of cancer cells at various stages during cancer progression.

[0081] 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 said gene is associated with the development of cancer.

[0082] The term "AXL (Tyrosine-protein kinase receptor UFO)" as used herein refers to a tyrosine kinase receptor present on the cell surface that mediates signal transduction involved in cell proliferation and survival. It is known to be involved in anticancer drug resistance in anticancer treatment. One embodiment of the AXL-targeted anticancer drug may be Bemcentinib or Gilteritinib.

[0083] The term "RET (Rearraged during transfection)" in this specification refers to a receptor that mediates signals involved in cell proliferation, cell death and survival, and RET mutation is known to be involved in cancer development. The RET target inhibitor may be, but is not limited to, selpercatinib or pralsetinib.

[0084] 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. The inhibitor targeting Braf may be Vemurafenib.

[0085] The term "PARP (Poly [ADP-ribose] polymerase)" in this specification refers to a protein that recognizes damaged DNA in the nucleus, activates it, 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 can be Olaparib, Talazoparib, Niraparib, or Rucaparib.

[0086] As used herein, "DNA methyltransferase (DNMT)" refers to an enzyme that adds a methyl group to histone proteins that wrap around DNA, and gene expression is suppressed through this process. The DMNT target inhibitor exhibits anticancer activity by inhibiting hypermethylation of tumor suppressor genes and inducing normal expression of the tumor suppressor genes. An embodiment of the DNMT target inhibitor may be azacitidine, decitabine, or guadecitabine.

[0087] The term "CDC (Cyclin dependent kinase) 4 / 6" as used herein refers to a protein that regulates the cell cycle and promotes cell growth, and is hyperactivated during the development and progression of various malignant tumors. CDK4 / 6 target inhibitors exhibit anticancer activity by inhibiting the cell cycle of cancer cells, suppressing cell proliferation, and inducing cell death. The CDK4 / 6 target inhibitor can be Abemaciclib or Palbociclib.

[0088] As used herein, "STING (Stimulator of Interferon The term "STING Genes" is 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. The STING agonist exhibits immune-enhancing effects and cancer angiogenesis-suppressing effects, and for example, the STING agonist can be CDNs, SB11285, DMXAA, etc.

[0089] As used herein, the term "antibody therapeutic agent" 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. The antibody therapeutic agent may be Cetuximab, Trastuzumab, Emtansine, Rituximab, Ibritumomab, Tositumomab, Brentuximab, Ofatumumab, Obinutuzumab, Necitumumab, Bevacizumab, Ramucirumab, Nivolumab, Pembrolizumab, Atezolizumab, Durvalumab, Ipilimumab, and the like.

[0090] The term "immune anticancer agent" as used herein refers to a substance that inhibits the activity of immune checkpoint proteins that suppress the differentiation, proliferation, and activity of immune cells, and is known to eliminate cancer cells by preventing the cancer cells from evading the immune system. The immune anticancer agent may be any one selected from the group consisting of anti-CTLA-4 antibody, anti-PD-1 antibody, anti-PD-L1 antibody, anti-PD-L2 antibody, anti-B7-H4 antibody, anti-HVEM antibody, anti-TIM3 antibody, anti-GAL9 antibody, anti-LAG3 antibody, anti-VISTA antibody, anti-KIR antibody, anti-BTLA antibody, and anti-TIGIT antibody. In one embodiment, the immune anticancer agent is ipilimumab (ip ilimumab, pembrolizumab, nivolumab, cemiplimab, atezolizumab, avelumab, and durvalumab.

[0091] The term "ADC (Antibody Drug Conjugate)" as used herein refers to a therapeutic agent that chemically binds an antibody to a cytotoxic drug and exhibits high anticancer effects through targeted delivery. The ADC may be Gemtuzumab-Ozogamicin, Brentuximab-Vedotin, Trastuzumab-Emtansine, Inotuzumab-Ozogamicin, Eribulin-Mesylate, etc.

[0092] 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), and guadecitabine. The therapeutic agent may be any one selected from the group consisting of adenomycin (DNMT), docetaxel, paclitaxel, pemetrexed, vemurafenib (Braf), abemaciclib (CDK4 / 6), cetuximab (EGFR), durvalumab (PD-L1), trastuzumab (Her2), DMXAA, and keytruda (PD-1).

[0093] The anti-cancer agent may include one or more anti-cancer agents. Specifically, the anti-cancer agent may be one or more selected from the group consisting of chemical anti-cancer agents, targeted anti-cancer agents, immune anti-cancer agents, and antibody therapeutic agents, and combinations thereof.

[0094] In one embodiment, the Lactobacillus plantarum GB104 strain can be used in combination with two or more different anticancer drugs (e.g., a first anticancer drug and a second anticancer drug). For example, the two anticancer drugs (e.g., a first anticancer drug and a second anticancer drug) can be a chemical anticancer drug and a targeted anticancer drug, a chemical anticancer drug and an anticancer virus, a targeted anticancer drug and an antibody therapeutic drug, a chemical anticancer drug and a cell therapeutic drug, or a chemical anticancer drug and an immune anticancer drug. It can also be a targeted anticancer drug and an anticancer virus, a targeted anticancer drug and an antibody therapeutic drug, a targeted anticancer drug and a cell therapeutic drug, or a targeted anticancer drug and an immune anticancer drug. It can also be an anticancer virus and an antibody therapeutic drug, an anticancer virus and a cell therapeutic drug, or an anticancer virus and an immune anticancer drug. It can also be an antibody therapeutic drug and a cell therapeutic drug, or an antibody therapeutic drug and an immune anticancer drug.

[0095] The Lactobacillus plantarum GB104 strain can be used together with three anti-cancer drugs, and the two anti-cancer drugs can be used together with a different anti-cancer drug.

[0096] The Lactobacillus plantarum GB104 strain can be used together with four anti-cancer drugs, and the three anti-cancer drugs can be used together with a different anti-cancer drug.

[0097] In one embodiment, the anti-cancer drug administered in combination with Lactobacillus plantarum strain GB104 may be one or more selected from the group consisting of 5-fluorouracil, anti-PD-1, bevacizumab, and cetuximab.

[0098] The Lactobacillus plantarum GB104 can be used in parallel with the anti-cancer drug and the anti-cancer vaccine, where the Lactobacillus plantarum GB104 strain and the anti-cancer drug are the same as above.

[0099] The term "anti-cancer vaccine" in this specification refers to an active immunotherapy that activates the immune system by administering tumor-specific antigens (TSA) 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 the antigen delivery method, and currently, cellular vaccines and DNA vaccines that are developed by introducing antigens are representative.

[0100] In one embodiment, the second active agent may comprise an anti-cancer agent as the sole active ingredient or may comprise one or more pharma- ceutically acceptable carriers, excipients or diluents.

[0101] In one embodiment, the anticancer agent can be prepared in a separate formulation for administration in combination with the composition of the present invention.The anticancer agent can be prepared as an injection containing the anticancer agent, preferably as a formulation administered by direct injection or perfusion intratumorally, intraarterially, intravenously, intravascularly, intrapleurally, intraperitoneally, intratracheally, intradurally, intramuscularly, endoscopically, intralesionally, percutaneously, subcutaneously, locally, stereotactically, or orally, but is not limited thereto.

[0102] In this specification, the term "containing as an active ingredient" means that a Lactobacillus strain, a homogenate of said strain, a culture medium, or an extract of the culture medium is added, and various ingredients are added as accessory ingredients for drug delivery, stabilization, etc., and this includes formulation in various forms.

[0103] In one embodiment, the first and second active agents may be co-administered simultaneously, sequentially or in reverse order.

[0104] The term "combination therapy" or "co-administration" or "in combination" as used herein refers 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, or at different administration frequencies, or via different routes, as appropriate.

[0105] Specifically, the combined administration may be such that the Lactobacillus plantarum strain and the anticancer drug are administered simultaneously, or the Lactobacillus plantarum strain is administered followed by the anticancer drug. The combined therapy according to the present invention may be defined as providing a synergistic effect when the efficacy, measured, for example, through the degree of response, the rate of response, the time to disease progression, or the survival time, is therapeutically superior to the efficacy that can be obtained by administering one or the other of the components of the combined therapy at a normal dose. For example, the efficacy of the combined therapy is synergistic when the efficacy is therapeutically superior to the efficacy obtained by using each of the components alone. In particular, the combined therapy is synergistic when it does not adversely affect one or more of the degree of response, the rate of response, the time to disease progression, and the survival time, in particular, does not adversely affect the duration of response, reduces / reduces or reduces problematic side effects compared to those occurring when each component is used at a normal dose, and reduces the normal dose of the Lactobacillus plantarum strain and the anticancer drug. If this can be achieved, it is believed that a synergistic effect will exist.

[0106] In one embodiment, the synergistic effect of the combined administration may be a reduction in the dose or frequency of administration of the existing anticancer drug. Specifically, when the Lactobacillus plantarum GB104 strain is combined with an anticancer drug (e.g., 5-fluorouracil), it can exhibit the same or similar anticancer activity at a lower dose or fewer doses than the dose of the anticancer drug (e.g., 5-fluorouracil) alone. In one embodiment, when the administration interval of the anticancer drug alone is 1 day, the anticancer activity and the Lactobacillus plantarum GB104 strain and the anticancer drug combined administration interval are 5 to 10 days (e.g., 7 days) are shown to be the same or similar. In one embodiment, the anticancer activity shown at a dose of 25 mg / kg when the anticancer drug alone is administered is the same or similar anticancer activity shown at a dose of 5 to 15 mg / kg (e.g., 10 mg / kg) when the Lactobacillus plantarum GB104 strain and the anticancer drug are administered together. Therefore, the combined administration of Lactobacillus plantarum GB104 has a synergistic effect that can reduce the usual dose and frequency of anticancer drugs.

[0107] 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.

[0108] The term "sequentially administered" as used herein is not particularly limited and means that the components of the combination therapy are not administered simultaneously, but rather 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, for example, from the range of 2 minutes to 96 hours, 1 day to 7 days, or 1 week, 2 weeks, or 3 weeks. In general, the time interval between administrations may range from a few minutes to a few hours, for example, from 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.

[0109] In one embodiment, the first active substance and the second active substance can be administered via independent routes. Each active ingredient can be administered independently by a person skilled in the art at an appropriate dosage and dose. Specifically, the Lactobacillus plantarum GB104 and the anti-cancer drug 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 Lactobacillus plantarum strain can be administered orally, and the anti-cancer drug can be administered intravenously or subcutaneously, but is not limited thereto.

[0110] Another aspect is to provide a pharmaceutical composition for preventing or treating cancer, comprising 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 extracts of the strain, the culture, and the disruptant, and a second active substance comprising an anticancer drug, or comprising the first active substance as an active ingredient and the second active substance being administered in combination.

[0111] The "strain", "first active substance", "anti-cancer agent", "second active substance", "anti-cancer activity", and "combined administration" are as defined above.

[0112] In one embodiment, the pharmaceutical composition can be administered to a mammal, including a human, by a variety of routes. The administration route may be any route that is commonly used, for example, oral, cutaneous, intravenous, intramuscular or subcutaneous, and preferably oral.

[0113] In one embodiment, the composition comprises a first oral formulation comprising a first active agent, and a second oral formulation comprising a second active agent, said first and second oral formulations being capable of being administered orally.

[0114] The term "prevention" herein can refer to any act of inhibiting or delaying the onset of a disease state in an individual by administration of a pharmaceutical composition according to an embodiment.

[0115] The term "treatment" 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 an embodiment.

[0116] According to one embodiment, the composition may contain 0.001% to 80% by weight 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 1000 mg, 1 mg to 100 mg, 0.01 mg to 1000 mg, 0.01 mg to 100 mg, 0.00 mg, or 0.01 mg to 100 mg. The strain is included in the composition in 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~ 1012 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~ 10 12 The composition may be included in the amount of CFU / g or as a culture of live or killed bacteria in the same number. Specifically, for adult patients, 1×10 3 ~1×10 16 CFU / g of live or killed bacteria can be administered once or in several doses. However, the dosage can be formulated in various ways depending on factors such as the formulation method, administration method, the patient's age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate and reaction sensitivity, and a person skilled in the art can appropriately control the dosage taking these factors into consideration. The number of administrations can be one or more than two times within the range of clinically acceptable side effects, and the administration can be administered to one or more sites. For animals other than humans, the dosage can be the same as that of humans per kg (body weight), or the dosage converted to the dosage based on, for example, the volume ratio (e.g., average value) of the organs (heart, etc.) of the target animal and human can be administered. Possible administration routes can include oral, sublingual, parenteral (e.g., subcutaneous, intramuscular, intraarterial, intraperitoneal, intradural, or intravenous), rectal, local (including transdermal), inhalation, and injection, or insertion of an implantable device or substance. In one embodiment, the subject of the treatment may be a human or other mammalian target, specifically, a human, a monkey, a mouse, a rat, a rabbit, a sheep, a cow, a dog, a horse, a pig, etc. According to one embodiment, the composition contains a killed dried strain, and can be administered at 1 g to 10 g, 0.5 g to 1.5 g, 2.5 g to 3.5 g, or 4.5 g to 5.5 g per dose, and can be administered once to three times a day.

[0117] 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 comprising 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 desires to obtain. A therapeutically effective amount of an anti-cancer agent may be determined based on the disease state, age, sex, etc. of the individual. and body weight, and the ability of the anti-cancer agent to elicit a desired response in the individual. A therapeutically effective amount is one in which any toxic or detrimental effects are outweighed by the therapeutically beneficial effects.

[0118] The pharmaceutical composition according to one embodiment may contain pharma- ceutically acceptable carriers and / or additives. For example, it may contain sterilized water, physiological saline, conventional buffers (such as phosphoric acid, citric acid, and other organic acids), stabilizers, salts, antioxidants (such as ascorbic acid), surfactants, suspending agents, isotonicity agents, or preservatives. For local administration, it may also contain combinations with organic substances such as biopolymers, inorganic substances such as hydroxyapatite, specifically collagen matrices, polylactic acid polymers or copolymers, polyethylene glycol polymers or copolymers, and chemical derivatives thereof.

[0119] 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 pharma- ceutically acceptable carrier, or may be frozen in a solution in which they are dissolved or dispersed.

[0120] The pharmaceutical composition according to one embodiment may appropriately contain a suspending agent, a solubilizing agent, a stabilizing agent, an isotonicity agent, a preservative, an anti-adsorption agent, a surfactant, a diluent, an excipient, a pH adjuster, a soothing agent, a buffer, a reducing agent, an antioxidant, etc., if necessary 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 the literature "Remington's Pharmaceutical Sciences, 19th ed., 1995". The pharmaceutical composition according to one embodiment may be prepared into a unit dose form by formulating it with a pharma- ceutical acceptable carrier and / or excipient according to a method that can be easily carried out by a person skilled in the art to which the present invention belongs, or may be prepared by putting it into a large-volume container. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or in the form of a powder, granules, tablets or capsules.

[0121] The pharmaceutical composition is administered in a pharma- ceutical effective amount. As used herein, the term "pharmaceutical effective amount" refers to an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to any medical treatment, and the effective dose level can be determined depending on factors including the type and severity of the patient's disease, the activity of the drug, sensitivity to the drug, administration time, administration route and excretion rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. The compositions of the present invention can be administered as individual therapeutic agents or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can obtain maximum effect at a minimum amount without side effects, which can be easily determined by those skilled in the art.

[0122] 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.

[0123] In one embodiment, the Lactobacillus plantarum GB104 strain was administered in combination with the anticancer drug 5-fluorouracil in a tumor animal model implanted with mouse colon cancer cell line MC-38, and showed an increased anticancer effect compared to the case where 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 tumor volume was reduced by 10% to 90% in the experimental group administered with GB104 in combination with the anticancer drug compared to the control group.

[0124] In one embodiment, when the Lactobacillus plantarum GB104 strain and a chemical anticancer agent were administered in combination, the colon cancer tumors in the negative control group, which were not administered the strain and the anticancer agent, were The present invention 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 100% tumor volume.

[0125] In one embodiment, the Lactobacillus plantarum GB104 strain was administered in combination with anti-PD-1, an immunological anticancer agent, to a tumor animal model implanted with mouse colon cancer cell line MC-38, and showed an increased anticancer effect compared to the administration of Lactobacillus plantarum GB104 strain alone. Specifically, the effect of suppressing the growth and progression of colon cancer tumors was confirmed. On average, the tumor volume was reduced by 10% to 90% in the experimental group administered GB104 in combination with an immunological anticancer agent compared to the control group.

[0126] In one embodiment, when the Lactobacillus plantarum GB104 strain is administered in combination with an immuno-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 the colon cancer tumor volume of 100% in a negative control group in which the strain and the anticancer agent were not administered.

[0127] In one embodiment, when the Lactobacillus plantarum GB104 strain was administered in combination with the targeted anticancer drug cetuximab or bevacizumab in a tumor animal model implanted with mouse colon cancer cell line MC-38, the anticancer effect was increased compared to the case where the Lactobacillus plantarum GB104 strain was administered alone. Specifically, the effect of suppressing the growth and progression of colon cancer tumors was confirmed. On average, the tumor volume was reduced by 10% to 90% in the experimental group administered with GB104 in combination with the targeted cancer drug compared to the control group.

[0128] In one embodiment, when the Lactobacillus plantarum GB104 strain is administered in combination with a targeted 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 in which the strain and the anticancer drug were not administered.

[0129] Another aspect is to provide a health functional food for preventing or ameliorating cancer, which contains 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 extracts of the strain, the culture, and the disrupted product, and a second active substance comprising an anticancer drug, or which contains the first active substance as an active ingredient and is administered in combination with the second active substance.

[0130] Another embodiment is a method for producing a composition comprising a first active substance comprising one or more selected from the group consisting of a Lactobacillus plantarum strain, a culture of said strain, a homogenate of said strain, or an extract of said strain, culture, or homogenate, and an anticancer agent. The present invention aims to provide a food composition for preventing or ameliorating cancer, which contains as an active ingredient a second active substance containing the first active substance, or which contains as an active ingredient the first active substance and is administered in combination with the second active substance.

[0131] The "strain", "first active substance", "anti-cancer agent", "second active substance", "anti-cancer activity", and "combined administration" are as defined above.

[0132] 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, for example, oral, cutaneous, intravenous, intramuscular or subcutaneous, and preferably oral.

[0133] In one embodiment, the composition comprises a first oral formulation comprising a first active agent, and a second oral formulation comprising a second active agent, said first and second oral formulations being capable of being administered orally.

[0134] In one embodiment, the health functional food may further comprise a food-based acceptable carrier.

[0135] As used herein, the term "food-safe" means exhibiting the property of being non-toxic to cells or humans exposed to the compound.

[0136] The term "improvement" in this specification can mean any action that at least reduces the parameters related to the condition being treated, such as the severity of symptoms. In this case, the health functional food can be used before or after the onset stage of the disease to prevent or improve cancer, either simultaneously or separately with a drug for treatment.

[0137] In the health functional food, 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 may be appropriately determined depending on the purpose of use (for prevention or improvement). In general, in the production of food or beverage, the health functional food may be added in an amount of about 15% by weight or less, more specifically about 10% by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health regulation, the amount may be less than the above range.

[0138] 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 carriers, diluents, excipients, and additives. Foods to which the compound according to one embodiment may be added include various foods, powders, granules, tablets, capsules, syrups, beverages, gums, teas, vitamin complexes, and health functional foods.

[0139] 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, gum acacia, 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.

[0140] In addition to containing the active ingredient, the health functional food may contain other ingredients as essential ingredients without any particular limitation. For example, it may contain various flavoring agents or can contain natural carbohydrates as additional components. Examples of the natural carbohydrates include monosaccharides such as glucose, fructose, etc. Disaccharides, such as maltose, sucrose, etc. Polysaccharides, e.g., conventional sugars such as dextrins, cyclodextrins, etc. It can be a sugar alcohol such as xylitol, sorbitol, erythritol, etc. As flavoring agents other than those mentioned above, natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycylrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of the natural carbohydrate can be appropriately determined by the selection of a person skilled in the art.

[0141] In addition to the above, the health functional food according to one embodiment may contain various nutrients, vitamins, minerals (electrolytes), flavorings such as synthetic flavorings and natural flavorings, coloring agents and enhancers (cheese, chocolate, etc.), 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.

[0142] 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 disease, an existing health functional food, or a newly developed health functional food.

[0143] When the health functional food contains other health functional foods having cancer prevention or improvement effects, it is important to mix them in amounts that can achieve maximum effect with the minimum amount without side effects, which can be easily determined by a person skilled in the art.

[0144] The food composition for preventing or improving cancer includes 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.

[0145] The compositions herein can be considered as food supplements. Food supplements, also known as dietary or nutritional supplements, can be considered as another specific pharmaceutical product. They are prepared to supplement the diet and provide nutrients or beneficial ingredients that are not available or cannot be obtained in sufficient amounts in the normal diet. Most food supplements are considered as foods, but sometimes they are considered as drugs, natural health products, or nutraceutical products. In the sense of the present invention, food supplements include health functional foods. Food supplements are usually sold over the counter without prescription. When food supplements take the form of pills or capsules, they contain the same excipients as those used in pharmaceuticals. However, food supplements can also take the form of food (e.g., infant formula) that is enriched with some nutrients. Thus, in a specific embodiment, the composition of the present invention is a food supplement.

[0146] The compositions according to the present invention can be administered as is, or mixed with a suitable edible liquid or solid, or can be in the form of tablets, pills, capsules, lozenges, granules, powders, suspensions, sachets, syrups, or freeze-dried in the form of unit doses, or in the form of monodoses of freeze-dried compositions that are mixed in a separate liquid container provided together prior to administration.

[0147] The composition of the present invention may be included in various edible foods and food products, such as milk products for infants. The term "edible product" in this specification is used in a broad sense to include any form of product that can be ingested by an animal in any way (e.g., a product that is received by the organs of the senses). The term "food product" is understood as an edible product that provides nutritional support to the body. In particular, 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 certain embodiments, the bacterial cells of the invention are homogenized with other ingredients such as cereals or powdered milk to form a manufactured infant formula.

[0148] Another aspect is to provide 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 disruptant of the strain, or extracts of the strain, the culture, and the disruptant, and a second active substance comprising an anticancer drug, or which comprises the first active substance as an active ingredient and is administered in combination with the second active substance.

[0149] 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, the culture, and the disruptant, and a second active substance comprising an anti-cancer drug, or the first active substance as an active ingredient in combination with the second active substance.

[0150] Another aspect is to provide 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 extracts of the strain, the culture, and the disruptant, and a second active substance comprising an anti-cancer drug, or a composition containing the first active substance as an active ingredient and the second active substance in combination.

[0151] The "strain", "first active substance", "anti-cancer agent", "second active substance", "anti-cancer activity", and "combined administration" are as defined above.

[0152] The individual may be an individual suffering from cancer. Further, the individual may be a mammal. , preferably a human.

[0153] The administration route, dosage and number of administrations of the Lactobacillus plantarum GB104 strain and the anticancer drug can be administered to the subject in various ways and amounts depending on the condition of the patient and the presence or absence of side effects, and the optimal administration method, dosage and number of administrations can be selected within an appropriate range by a person skilled in the art. In addition, in addition to the active ingredient, the active ingredient may be administered in combination with other drugs (e.g., the above-mentioned anticancer drugs) or physiologically active substances known to have a therapeutic effect on cancer diseases, or may be formulated in the form of a combination preparation with other drugs. Effect of the Invention

[0154] The Lactobacillus plantarum strain or a culture medium of said strain can not only inhibit the proliferation of cancer cells or induce the death of cancer cells, but also exhibit a synergistic effect in inhibiting the proliferation of cancer cells and tumors when administered in combination with an anticancer drug, and can be usefully used in the form of a pharmaceutical composition or health functional food for the prevention, treatment or improvement of cancer. [Brief description of the drawings]

[0155] [Figure 1] FIG. 1 is a graph showing the results of analyzing the survival rate of cells after treating the human colon cancer cell line HCT116 with culture supernatants of bacterial strains different from GB104. [Diagram 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 supernatant of GB104 and a control strain (WCFS1). [Diagram 3] FIG. 3 is a graph showing the effect of cancer cell death 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 tumor formation inhibitory effect of treating mouse MC-38 colon cancer cell line, LLC1 lung cancer cell line, and 4T1 breast cancer cell line with GB104 culture supernatant. [Diagram 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 a comparative analysis of tumor volume 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 combined 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 after administration of GB104 and anti-PD-1 alone or in combination 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 of GB104 and bevacizumab 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 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 of 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 after single administration and combined administration of GB104 and Cetuximab in a xenografted HT-29 mouse colon carcinoma model. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0156] In the following, preferred embodiments are presented 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, as various modifications may be made to the embodiments, and the embodiments are not limited to the embodiments disclosed below, and may be implemented in various forms.

[0157] Embodiment 1. Isolation and identification of Lactobacillus plantarum GB104 strain

[0158] Isolation and identification of Lactobacillus plantarum GB104 strain were performed according to the methods described in Korean Patent Application No. 10-2020-0186738 and Korean Patent Application No. 10-2022-0080567, the entireties of which are incorporated herein by reference.

[0159] Briefly, Lactobacillus plantarum GB104 was isolated from vaginal samples of healthy women who visited the hospital for health checkups. First, vaginal samples were collected with a cotton swab, pre-inoculated onto Rogosa SL (MRS) plates, and 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 pure isolation, strain culture was performed using MRS medium. Next, among the cultured strains, Lactobacillus plantarum GB104 strains with low fat cell accumulation inhibitory effects and cytotoxicity were finally selected. To identify the finally 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, and the 16s rRNA sequence of Lactobacillus plantarum GB104 is shown in SEQ ID NO:1. The present inventors named GB104 bacterium "Lactobacillus plantarum GB104" (deposit number: KCTC14107BP) and deposited it at 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 described interchangeably.

[0160] 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 In this experimental example, a human colon cancer cell line was treated with various L. plantarum culture supernatants, including the L. plantarum GB104 strain, and the cell viability was screened by MTT assay.

[0161] Human colon cancer cell line HCT116 cells were placed in a 96-well plate at 2x10 3 After dispensing the cells into each well and culturing for 24 hours, the culture supernatant of various L. plantarum strains was treated at a concentration of 10% and cultured at 37 °C and 5% CO2 for 72 hours. The culture supernatant was prepared by culturing L. plantarum strains in MRS medium, precipitating the strains by centrifugation, collecting only the supernatant, and filtering it through a 0.22 μm filter to obtain the supernatant. After treating the MTT solution to 0.5 mg / mL using Cell Growth Kit I (MTT) (Roche), the cells were further cultured for 4 hours, and the purple formazan crystals produced by living cells due to metabolic activity were dissolved in the solubilization solution, and the absorbance was measured at 570 nm, as shown in Figure 1.

[0162] FIG. 1 is a graph showing the results of analyzing the survival rate of cells after treating the human colon cancer cell line HCT116 with culture supernatants of bacterial strains different from GB104.

[0163] 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 the cancer cell viability by approximately 95%, confirming that it was the most effective in inhibiting cancer cell proliferation.

[0164] 1.2. Confirmation of viability of human cancer cell lines by treatment with GB104 culture supernatant In this experimental example, various human somatic cancer cell lines were treated with the L. plantarum GB104 culture supernatant, and the cell viability was screened by MTT assay.

[0165] 0.8-1.5×10 human cancer cell lines in a 96-well plate 4 The cells were dispensed into each well and cultured for 24 hours, after which the culture supernatant of L. plantarum GB104 strain was treated at a concentration of 10% and cultured at 37°C and 5% CO2 for 48 hours. The cell viability was measured in the same manner as in Experimental Example 1.1 above, and the results are shown in Table 1.

[0166] [Table 1] As a result, we confirmed that L. plantarum GB104 culture supernatant reduced cell viability in all treated human cancer cell lines compared to the untreated control (MRS), although cell viability varied depending on the characteristics of the cancer cells.

[0167] 1.3. Confirmation of cancer cell cycle by treatment with GB104 culture supernatant In this experimental example, a human colon cancer cell line was treated with L. plantarum GB104 culture supernatant, and changes in the cancer cell cycle were confirmed using a flow cytometer.

[0168] Human colon cancer cell line HCT116 cells were placed in a 6-well plate at 5x10 4 After dispensing the cells into each well and culturing for 24 hours, the culture supernatant of the L. plantarum GB104 strain was treated at a concentration of 10% and cultured at 37°C and 5% CO2 for 48 hours. The culture supernatant was prepared by culturing the L. plantarum strain in MRS medium, precipitating the strain by centrifugation, and collecting only the supernatant, which was then filtered through a 0.22 μm filter to obtain the supernatant. After 48 hours, the cells were detached using trypsin-EDTA, then centrifuged to recover the cells, and then diluted with 70% ethanol. The cells were fixed with PBS and stored at 4°C for at least 1 hour. After treatment with RNase A and staining with PI (propidium iodine), the extent of cancer cell death was confirmed by DNA content analysis using a flow cytometer (BD FACSymphony A3 Cell Analyzer), and the results are shown in Figure 2.

[0169] 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 supernatant of GB104 and a control strain (WCFS1).

[0170] As shown in Figure 2, the sub-G1 region of cells treated with L. plantarum GB104 culture supernatant was increased by approximately 9.8-fold compared to the control cells treated with MRS, confirming the fact that GB104 induces the death of cancer cells.

[0171] 1.4.Early and late stage cancer cell death effects of GB104 culture supernatant treatment in human cancer cell lines In this experimental example, L. plantarum GB104 culture supernatant was treated with a human cancer cell line, and the cancer cell killing effect was confirmed using a flow cytometer.

[0172] 0.5-1x10 human cancer cell lines in a 6-well plate 5 After dispensing the cells into each well and culturing for 24 hours, the culture supernatant of L. plantarum GB104 strain was treated at a concentration of 10% and cultured at 37℃ and 5% CO2 for 24 hours. The culture supernatant was obtained by culturing L. plantarum strains in MRS medium, precipitating the strain by centrifugation, collecting only the supernatant, and filtering it through a 0.22μm filter. After 24 hours, the cells were removed by trypsin-EDTA treatment and centrifuged to recover the cells. The cells were mixed with 100μL binding solution using FITC Annexin V Apoptosis Detection Kit with 7-AAD (BioLegend), and then 5μL FITC Annexin V and 5μL 7-AAD were added and reacted at room temperature for 15 minutes in the dark. Fluorescence measurements to confirm the death of cancer cells were performed using a flow cytometer (BD FACSymphony A3 Cell Analyzer), and the results are shown in Figure 3. Normal living cells are not labeled with annexin V or 7-AAD, while cells in the early stage of cell death are labeled with annexin V but not with 7-AAD. Cells in the late stage of cell death can be distinguished and confirmed as being labeled with both annexin V and 7-AAD.

[0173] FIG. 3 is a graph showing the effect of cancer cell death 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).

[0174] As shown in Figure 3, L. plantarum GB104 culture supernatant confirmed a significant increase in early and late cell death in treated colon and breast cancer cell lines compared to the untreated control (MRS).

[0175] This confirmed the fact that GB104 is involved in inducing the death of cancer cells. Experimental Example 2. Inhibitory effect of GB104 culture supernatant on tumor formation In this embodiment, the tumor formation suppression effect of treatment with L. plantarum GB104 strain was confirmed in various mouse cancer cell lines.

[0176] The culture supernatant was prepared by culturing L. plantarum in MRS medium, precipitating the bacteria by centrifugation, and then collecting the supernatant, which was then filtered through a 0.22 μm filter. Specifically, mouse colon cancer cell line MC-38, mouse lung cancer cell line LLC1, and mouse breast cancer cell line 4T1 were each cultured in a 6-well plate at 0.5–2 × 10 3 To become a cell The cells were diluted in 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 new medium, and treated with 1% culture supernatant of L. plantarum GB104 strain. The medium was replaced with new medium containing the culture supernatant of the strain at intervals of 2 to 3 days, and cultured for 7 days. The culture plate was washed twice with DPBS and fixed by treating with 4% formalin. The fixed cells were washed twice with DPBS, stained with 0.5% crystal violet solution for 5 minutes, and then washed with distilled water to confirm their colony forming ability. The results are shown in Figure 4.

[0177] FIG. 4 is a graph showing the tumor formation inhibitory effect of treating mouse MC-38 colon cancer cell line, LLC1 lung cancer cell line, and 4T1 breast cancer cell line with GB104 culture supernatant.

[0178] As shown in Figure 4, although the ability to suppress tumor formation varies depending on the characteristics of the cancer cells, it was confirmed that L. plantarum GB104 culture supernatant suppressed tumor formation in all treated mouse cancer cell lines compared to the untreated control group (MRS).

[0179] Experimental Example 3. Confirmation of tumor growth inhibition effect by combined administration of GB104 strain and chemical anticancer drug 5-fluorouracil In this experiment, the tumor treatment effect of the combined administration of L. plantarum GB104 strain and 5-fluorouracil (5-FU) was confirmed in a colon carcinoma MC-38 allograft model. Six-week-old c57BL / 6 mice were brought into the warehouse for a one-week purification 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 × 10 5 Tumor models were established by subcutaneous injection of 100 μL of cells per 100 μL. Tumor size was measured using a digital caliper, and tumor volume (mm 3 )=(width 2 The tumor size was calculated by the formula: tumor size was calculated by dividing the tumor size by the tumor cell size (x length) / 2. Only mice with tumor sizes falling within a certain range were selected on the 5th day after tumor cell injection, and the tumor sizes of each group were classified so that the average tumor size was equal. L. plantarum GB104 strain was then added to the animal model at a dose of 1 × 10 per mouse. 9 CFU were orally administered daily from day 6 until 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.

[0180] 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.

[0181] FIG. 6 is a graph showing the results of a comparative analysis of tumor volume after single administration and combined administration of GB104 and 5-FU in an allografted MC-38 mouse colon carcinoma model.

[0182] 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 (Control). In addition, a significant reduction in tumor size due to a synergistic effect was observed in the group administered with L. plantarum GB104 and 5-FU in combination compared to the groups administered with L. plantarum GB104 or 5-FU alone.

[0183] Experimental Example 4. Confirmation of tumor growth suppression effect by combined administration of GB104 strain and immune anti-cancer drug anti-PD-1 In this experiment, we confirmed the tumor treatment 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 brought into the laboratory for a one-week purification 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 × 10 5 Tumor models were established by subcutaneous injection of 100 μL of cells per 100 μL. Tumor size was measured using a digital caliper, and tumor volume (mm 3 ) =(width 2 The tumor size was calculated by (x length) / 2. The tumor size was 10-30 mm on the 5th day after tumor cell injection. 3 Only mice falling within the range were selected and randomly assigned to each group, and L. plantarum GB104 strain was then added to the animal model at 1 × 10 per mouse. 9 CFU were orally administered daily from the 5th day until just before the end of the study. The immunotherapy anti-cancer drug anti-PD-1 (5 mg / kg) was intraperitoneally administered twice a week either alone or in combination with GB104.

[0184] FIG. 7 is a graph showing the results of analyzing the synergistic effect by measuring tumor volume after combined administration of GB104 and anti-PD-1 in an allografted MC-38 mouse colon carcinoma model.

[0185] FIG. 8 is a graph showing the results of a comparative analysis of tumor volume after administration of GB104 and anti-PD-1 alone or in combination in an allografted MC-38 mouse colon carcinoma model.

[0186] As shown in Figures 7 and 8, compared to the negative control group (Control), a significant reduction in tumor size due to a synergistic effect was observed in the group administered L. plantarum GB104 in combination with anti-PD-1 compared to the groups administered L. plantarum GB104 or anti-PD-1 alone.

[0187] Experimental Example 5. Confirmation of tumor growth inhibition effect by combined administration of GB104 strain and targeted anticancer drug bevacizumab In this experimental example, the tumor treatment effect of the combined administration of L. plantarum GB104 strain and bevacizumab (anti-VEGF) was confirmed in a xenograft model using immunodeficient mice. 6-week-old NOG mice were brought into the warehouse for a one-week purification period, and the right flank area was depilated before the experiment was started at 7 weeks of age. The human colon cancer cell line HT-29 was inoculated into the right flank of each mouse at 3 × 10 6 Tumor models were established by subcutaneous injection of 100 μL of cells per 100 μL. Tumor size was measured using a digital caliper, and tumor volume (mm 3 )=(width 2 On the third day after tumor cell injection, only mice with tumor sizes falling within a certain range were selected and classified so that the average tumor size of each group was equal. L. plantarum GB104 strain was then injected into the animal model at 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.

[0188] FIG. 9 is a graph showing the results of analyzing synergistic effects by measuring tumor volume after combined treatment of GB104 and bevacizumab in a xenografted HT-29 mouse colon carcinoma model.

[0189] FIG. 10 is a graph showing the results of a comparative analysis of tumor volume after single administration and combined administration of GB104 and bevacizumab in a xenografted HT-29 mouse colon carcinoma model.

[0190] As shown in Figures 9 and 10, it was confirmed that 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 in combination with bevacizumab compared to the group administered with L. plantarum GB104 or bevacizumab alone.

[0191] Experimental Example 6. Confirmation of tumor growth inhibition effect by combined administration of GB104 strain and targeted anticancer drug cetuximab In this experiment, we investigated the efficacy of combined administration of Lactobacillus plantarum GB104 and cetuximab (anti-EGFR) in tumor treatment in a xenograft model using immunodeficient mice. The results were confirmed. Six-week-old NSG mice were brought into the warehouse, and after a one-week purification period, the right flank area was depilated and the mice were then aged 7 weeks to proceed with the experiment. The human colon cancer cell line HT-29 was inoculated into the right flank of each mouse at 3 × 10 6 Tumor models were established by subcutaneous injection of 100 μL of cells per 100 μL. Tumor size was measured using a digital caliper, and tumor volume (mm 3 )=(width 2 On the third day after tumor cell injection, only mice with tumor sizes falling within a certain range were selected and classified so that the average tumor size of each group was equal. L. plantarum GB104 strain was then injected into the animal model at 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 cetuximab (40 mg / kg) was intraperitoneally administered twice weekly either alone or in combination with GB104.

[0192] FIG. 11 is a graph showing the results of analyzing synergistic effects by measuring tumor volume after combined treatment of GB104 and Cetuximab in a xenografted HT-29 mouse colon carcinoma model.

[0193] FIG. 12 is a graph showing the results of a comparative analysis of tumor volume after single administration and combined administration of GB104 and Cetuximab in a xenografted HT-29 mouse colon carcinoma model.

[0194] As shown in Figures 11 and 12, a significant tumor growth inhibitory effect was confirmed in the group administered L. plantarum GB104 compared to the negative control group (Control). Furthermore, a synergistic effect of reducing tumor size was confirmed in the group administered L. plantarum GB104 in combination with cetuximab compared to the group administered cetuximab alone. [Accession number]

[0195] Name of depository institution: Korea Institute of Bioscience and Biotechnology Accession number: KCTC14107BP Date of acceptance: 20200114

Claims

1. The composition 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 anticancer agent; or A pharmaceutical composition for preventing or treating cancer, comprising the first active substance as an active ingredient, and the second active substance is 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 activity.

3. The pharmaceutical composition for preventing or treating cancer according to claim 1 , wherein the first active substance induces cell death in cancer cells.

4. The pharmaceutical composition for preventing or treating cancer according to claim 1 , wherein the strain comprises the 16S rRNA sequence 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 anticancer agent comprises a first anticancer agent and a second anticancer agent, the first and second anticancer agents being different from each other.

8. The pharmaceutical composition for preventing or treating cancer according to claim 1 , wherein the anticancer agent is any one selected from the group consisting of chemical anticancer agents, targeted anticancer agents, and immune anticancer agents.

9. 9. The pharmaceutical composition for preventing or treating cancer according to claim 8, wherein the anticancer chemical agent is any one selected from the group consisting of an alkylating agent, a microtubule inhibitor, an antimetabolite and a topoisomerase inhibitor.

10. 9. The pharmaceutical composition for preventing or treating cancer according to claim 8, wherein the anticancer chemical 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, bepcid, paclitaxel, bleosane, adriamycin and cervidin.

11. The targeted anticancer drug is 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. The pharmaceutical composition for preventing or treating cancer according to claim 8 , which targets any one of the proteins selected from the above.

12. The targeted anticancer drugs include abciximab, adalimumab, basiliximab, bezlotoxumab, canakinumab, daclizumab, denosumab, efalizumab, golimumab, inflectra, natalizumab, olaratumab, and omalizumab.

9. The pharmaceutical composition for preventing or treating cancer according to claim 8, which is any one selected from the group consisting of palivizumab, panitumumab, trastuzumab, pertuzumab, cetuximab, tocilizumab, secukinumab, ustekinumab, bevacizumab, avelumab, and DMXAA.

13. 9. The pharmaceutical composition for preventing or treating cancer according to claim 8, wherein the immunological anticancer agent 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 (CD 11 a / CD 18), LIGHT, NKG2C, NKp80, OX40, PD-1, PD-L1, PD-L2, SLAMF7, TGFRp, TIGIT, Tim3 and VISTA.

14. The immunological anticancer drug is atezolizumab, avelumab, durvalumab, nivolumab, pembrolizumab, abagovomab, adecatum, afutuzumab, alemtuzumab, anatumomab mafenatox, apolizumab, blinatumomab, BMS-936559, catumaxomab, cemiplimab, epacadostat, epratuzumab, indoximod, inotuzumab, ozogamicin, intelmumab, ipirlimumab, isatuximab, lambrolizumab, MED 14736, MPDL3280A, obinutuzumab, okaratuzumab, ofatumumab, olaratumab, visilizumab, rituximab, ticilimumab, samalizumab, and tremelimumab. The pharmaceutical composition for preventing or treating cancer according to claim 8, wherein the compound is any one selected from the group consisting of 14736, MPDL3280A, obinutuzumab, okaratuzumab, ofatumumab, olaratumab, visilizumab, rituximab, ticilimumab, samalizumab, and tremelimumab.

15. 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, esophageal cancer, head and neck cancer, skin cancer, small intestine cancer, anal cancer, colon cancer, rectal cancer and lymphoma.

16. The pharmaceutical composition for preventing or treating cancer according to claim 15, wherein the colon cancer is any cancer selected from the group consisting of ascending colon, transverse colon, descending colon, sigmoid colon and rectal mucosa.

17. 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.

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

19. 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.

20. The composition 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 anticancer agent; 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.

21. The health functional food for preventing or improving cancer according to claim 20, wherein the strain is the strain deposited under accession number KCTC14107BP.

22. The health functional food for preventing or ameliorating cancer according to claim 20, wherein the anticancer drug comprises a first anticancer drug and a second anticancer drug, the first and second anticancer drugs being different from each other.

23. The health functional food for preventing or ameliorating cancer according to claim 20, wherein the anticancer agent is any one selected from the group consisting of chemical anticancer agents, targeted anticancer agents, and immune anticancer agents.

24. The health functional food for preventing or improving cancer according to claim 20, 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, esophageal cancer, head and neck cancer, skin cancer, small intestine cancer, anal cancer, colon cancer, rectal cancer and lymphoma.

25. The health functional food for preventing or ameliorating cancer according to claim 24, wherein the colon cancer is any one selected from the group consisting of ascending colon, transverse colon, descending colon, sigmoid colon and rectal mucosa.

26. The health functional food for preventing or ameliorating cancer according to claim 20, wherein the first active substance and the second active substance are administered simultaneously, sequentially or in reverse order.

27. The health functional food for preventing or ameliorating cancer according to claim 20, wherein the first active substance and the second active substance are administered orally, intravenously or subcutaneously.

28. The health functional food for preventing or ameliorating cancer according to claim 20, wherein the first active substance is administered orally and the second active substance is administered intravenously or subcutaneously.

29. 1. A method for preventing or treating cancer, comprising administering to an individual in need thereof a first active agent comprising an effective amount of a Lactobacillus plantarum strain, a culture of said strain, a homogenate of said strain, or a mixture thereof, and a second active agent comprising an effective amount of an anti-cancer agent.

30. A composition 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 anticancer agent as an active ingredient, for the manufacture of a pharmaceutical preparation for preventing or treating cancer, or a composition comprising the first active substance as an active ingredient, Use of the composition in which said second active agent is co-administered.

31. 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 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 ameliorating cancer.

Images