Composition containing Lactobacillus reuteri ATG-F4 strain as an effective ingredient for antitumor or tumor metastasis inhibition

Abstract

The present invention relates to a composition for combined administration comprising the Lactobacillus reuteri ATG-F4 (accession number KCTC13717BP) strain and a chemical anticancer agent, and it is confirmed that tumor metastasis is significantly inhibited and the growth of the tumor itself is reduced through the combined administration of the strain and the anticancer agent. In addition, in the generated tumors, it inhibited the cancer metastasis factors IL-6 receptor (Interleukin-6 receptor) and TGF-β (transforming growth factor-β), increased the cell death factors BAX (Bcl-2 associated X Protein) and Caspase-3 fragment proteins, and significantly reduced the expression of tumor growth factors such as AKT (Akt8 virus transforming kinase target), ERK1 / 2 (Extracellular signal-regulated kinases 1 / 2), MEK1 (MAPK / ERK kinase 1), and COX-2 (Cyclooxygenase-2). These results suggest that by combining the Lactobacillus reuteri ATG-F4 strain with the above-mentioned chemical anticancer agent, it is possible to use a highly effective anticancer agent or metastasis inhibitor.

Description

Technology Field

[0001] The present invention relates to Lactobacillus reuteri ATG-F4 ( Lactobacillus reuteri The present invention relates to a composition for anti-tumor or tumor metastasis inhibition characterized by containing the strain ATG-F4 (accession number KCTC13717BP) as an active ingredient. Background Technology

[0002] Cancer, a malignant tumor that threatens human health, ranks first or second globally as a cause of death and is the most common cause of death in Western societies, second only to cardiovascular disease. Lung cancer is on the rise due to the aging population, increased smoking rates, and air pollution in modern society. Furthermore, colorectal, breast, and prostate cancers are on a continuous upward trend due to the consumption of high-fat diets resulting from the Westernization of dietary habits, increased exposure to environmental pollutants, and increased alcohol consumption. Therefore, there is an urgent need to create anticancer substances that can enable the early prevention and treatment of cancer, thereby contributing to the improvement of human health, the enhancement of the quality of a healthy life, and the advancement of public health.

[0003] The commonly used anticancer drugs are cytotoxic agents that exert their anticancer effects through a mechanism in which they directly attack and kill cancer cells. Major mechanisms of action include the induction of DNA damage, inhibition of cell division, and metabolic antagonism. These anticancer effects are confirmed by changes in apoptosis factors (BAX, Caspase-3 fragment) and tumor growth factors (AKT, ERK1 / 2, MEK1, COX-2).

[0004] Furthermore, cancer cells detach from the primary site and migrate to other organs via blood vessels or lymphatic vessels to form new tumors; this process is known as tumor metastasis. This process is divided into several stages—local invasion, intravascular invasion, circulation, and establishment at metastatic sites—each of which is regulated by the interactions of various molecules and cells. IL-6 and TGF-β are cytokines involved in inflammation and immune responses that play crucial roles in the tumor microenvironment; they are involved in promoting metastasis by regulating the behavior of cancer cells at various stages of the process.

[0005] IL-6 promotes the proliferation, survival, invasion, and angiogenesis of cancer cells, and induces epithelial-mesenchymal transition (EMT), thereby increasing the motility and invasiveness of cancer cells. Furthermore, it plays a role in facilitating cancer cell metastasis by promoting ECM degradation and angiogenesis. TGF-β initially exhibits tumor-suppressing effects, but as cancer progresses, it displays tumor-promoting effects and promotes metastasis through EMT induction, promotion of cancer cell invasion and angiogenesis, immunosuppression, and the maintenance of cancer stem cells. IL-6 and TGF-β interact and exhibit synergistic effects, making them key factors that can further promote tumor metastasis. In other words, if either IL-6 or TGF-β is blocked, an anti-metastatic effect capable of inhibiting cancer cell metastasis can be expected. Prior art literature

[0006] Republic of Korea Registered Patent No. 10-1951919 (Title of Invention: Novel Lactobacillus reuteri ATG-F4 strain having dopamine secretion-enhancing function, composition for the prevention or treatment of mental disorders containing the same, Applicant: A2Gen Co., Ltd., Registration Date: Feb. 19, 2019) Republic of Korea Registered Patent No. 10-2049700 (Title of Invention: Composition for the prevention or treatment of muscle diseases containing Lactobacillus reuteri ATG-F4, Applicant: A2Gen Co., Ltd., Registration Date: November 21, 2019) The problem to be solved

[0007] The objective of the present invention is Lactobacillus reuteri ATG-F4 ( Lactobacillus reuteri The present invention provides a composition for anti-tumor or tumor metastasis inhibition characterized by containing the strain ATG-F4 (accession number KCTC13717BP) as an active ingredient. means of solving the problem

[0008] The present invention relates to Lactobacillus reuteri ATG-F4 ( Lactobacillus reuteri The present invention relates to a composition for anti-tumor or tumor metastasis inhibition characterized by containing the strain ATG-F4 (accession number KCTC13717BP) as an active ingredient.

[0009] The strain may include one or more selected from the group consisting of the cell, the culture of the strain, the extract, concentrate, and dried product of the culture.

[0010] The above strain inhibits the protein expression of the tumor metastasis factor IL-6 receptor (Interluekin-6 receptor) or TGF-β (transforming growth factor-β), increases the expression of the apoptosis factor BAX (Bcl-2 associated X Protein) and Caspase-3 fragment protein, and increases the protein expression of the tumor growth factors AKT (Akt8 virus transforming kinase target), ERK1 / 2 (Extracellular signal-regulated kinases 1 / 2), MEK1 (MAPK / ERK kinase 1), and COX-2 (Cyclooxygenase-2).

[0011] The strain may induce an increase in the production of butyric acid (BA). The butyric acid may be produced due to changes in the intestinal microbial flora of an individual administered Lactobacillus reuteri ATG-F4, and preferably may be induced by an increase in the butyricicoccus flora.

[0012] The increase in the production of the above-mentioned butyric acid is confirmed by changes in blood concentration in the cecum or blood of the individual to whom the above-mentioned strain was administered, and the said individual may be any human or mammal other than a human, or any animal capable of developing tumors.

[0013] The above composition is characterized by the co-administration of a chemical anticancer agent. Preferably, the anticancer agent is selected from the group consisting of oxaliplatin, cisplatin, cyclophosphamide, ifosfide, bendamustine, melphalan, carboplatin, busulfan, dacarbazine, temozolomide, fluorouracil, capecitabine, doxifluridine, tegafur, cytarabine, azacitidine, decitamine, enositabine, methotrexate, pemetrexid, pralatrexate, cladribine, cloparabine, fludarabine, mercaptopurine, irinotecan, doxorubicin, daunorubicin, epirubicin, idarubicin, mitoxantrone, etoposide, topotecan, cabazitaxel, paclitaxel, docetaxel, vinblastine, vincristine, vinorelbine, bleomycin, hydroxyurea, and mitomycin C. One or more types may be selected.

[0014] Accordingly, the present invention can provide a pharmaceutical composition for anticancer or antimetastatic use containing the above-mentioned Lactobacillus reuteri ATG-F4 strain and a pharmaceutical excipient.

[0015] The present invention will be described in detail below.

[0016] The above anticancer agents are cytotoxic chemotherapy agents that exert an anticancer effect by directly attacking cancer cells that differentiate more rapidly than normal cells; their types include alkylating agents (oxaliplatin, cisplatin, cyclophosphamide, ifosfide, bendamustine, melphalan, carboplatin, busulfan, dacarbazine, temozolomide), metabolic antagonists (fluorouracil, capecitabine, doxifluridine, tegafur, cytarabine, azacitidine, decitamine, enositabine, methotrexate, pemetrexid, pralatrexate, cladribine, cloparabine, fludarabine, mercaptopurine), DNA twirlase inhibitors (irinotecan, doxorubicin, daunorubicin, epirubicin, idarubicin, mitoxantrone, etoposide, topotecan), and microtubules There are inhibitors (carbazitaxel, paclitaxel, docetaxel, vinblastine, vincristine, vinorelbine) and others (bleomycin, hydroxyurea, mitomycin C), and the above chemotherapy drugs have similar enzymatic mechanisms of action, mainly through the inhibition of DNA damage or DNA synthesis.

[0017] The above antitumor composition or anticancer agent may be a therapeutic agent targeting the following cancers. Preferably, the cancer may be a disease selected from the group consisting of hemangioma, angiofibroma, lung cancer, non-small cell lung cancer, liver cancer, colon cancer, bone cancer, pancreatic cancer, skin cancer, head cancer, neck cancer, melanoma, uterine cancer, ovarian cancer, rectal cancer, stomach cancer, pro-anal cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, bladder cancer, kidney or ureteral cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system tumor, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma and pituitary adenoma, and is not limited to other cancer diseases.

[0018] The above 1.0 x 10 8 Up to 4.0 x 10 10Based on the administration of the ATG-F4 strain in CFU, more preferably 1.0 x 10 9 Up to 4.0 x 10 10 Based on the ATG-F4 strain administration rate in CFU, most preferably 1.0 x 10 10 Up to 4.0 x 10 10 Based on the administration of the ATG-F4 strain in CFU, additionally, based on the co-administration of an anticancer drug at 10–100 mg / kg, more preferably based on the co-administration of an anticancer drug at 30–100 mg / kg, and most preferably based on the co-administration of an anticancer drug at 50–100 mg / kg, tumor metastasis is inhibited by more than 89% (see Fig. 2). In addition, when the strain and the anticancer drug are administered in combination, the tumor weight is reduced by more than 50% compared to the administration of the anticancer drug alone (see Fig. 3).

[0019] Meanwhile, under these conditions, compared to the administration of the anticancer drug alone, BAX protein expression increased 3.5 to 5.5 times in the group administered in combination with the anticancer drug and the ATG-F4 strain, and Caspase-3 fragment protein increased 3.0 to 8.5 times. Phosphorylation of AKT decreased by 65 to 95% in the group administered in combination with the ATG-F4 strain compared to the administration of the anticancer drug alone, which is a result that is significantly suppressed compared to the values ​​in the tumorigenic group. Under the same criteria, phosphorylation of ERK and MEK was suppressed by 45 to 70% in the group administered in combination with the ATG-F4 strain compared to the group administered in combination with the anticancer drug alone, and expression of COX2, IL-6 receptor TGF-β was suppressed by 40 to 70% in the group administered in combination with the ATG-F4 strain compared to the group administered in combination with the anticancer drug alone.

[0020] In the present invention, "culture of a strain" includes a culture medium, such as the culture solution itself cultured in a liquid medium, and the supernatant (filtrate) obtained by filtering and / or centrifuging the culture solution to remove the strain.

[0021] In the present invention, "extracts, concentrates, and dried products of a culture" may undergo centrifugation or filtration processes to remove the liquid culture medium from the culture and recover only the concentrated cells, but the present invention is not limited thereto. Additionally, the concentrated cells may be preserved by drying, freezing, or freeze-drying according to conventional methods so as not to lose their activity.

[0022] The above Lactobacillus reuteri ATG-F4 strain, the cells of the strain, the culture of the strain, the extract, concentrate, and dried product of the culture may be added to the composition of the present invention in an amount of 0.001 to 100 weight%.

[0023] In the present invention, "anticancer chemotherapy" refers to a systemic treatment method that prevents or kills cancer cells by using drugs, namely anticancer agents, for cancer treatment.

[0024] In an embodiment of the present invention, a process is presented in which anticancer or tumor metastasis-inhibiting efficacy is exerted through chemotherapy in which Lactobacillus reuteri ATG-F4 and an anticancer agent are administered in combination. To this end, the growth of cancerous tumors and metastatic tumors is inhibited by oral administration of Lactobacillus reuteri ATG-F4 and the administration of the combination anticancer agent FOLFOX to mice with induced cancer. By using the combination anticancer agent, it is suggested that the simultaneous use of two or more agents along with the administration of Lactobacillus reuteri ATG-F4 is effective for anticancer or tumor metastasis inhibition.

[0025] In other words, while anticancer or anti-metastatic efficacy is confirmed upon administration of conventional anticancer drugs, there are limitations in that cytotoxic anticancer drugs cannot inhibit the growth of cancer cells or completely suppress metastasis. When Lactobacillus reuteri ATG-F4 is administered in combination with the aforementioned anticancer drug, the expression of apoptosis-related proteins increases, while proteins related to tumor growth / proliferation and tumor metastasis decrease, thereby reducing or decreasing tumor growth. Consequently, Lactobacillus reuteri ATG-F4 acts as a 'booster,' 'adjuvant,' or 'amplifier' that promotes or enhances the anticancer or anti-metastatic efficacy of the existing anticancer drug. This may also be applicable to various individuals that have developed resistance to various anticancer drugs.

[0026] In addition, the present invention provides a pharmaceutical composition for anticancer or antimetastatic use containing the Lactobacillus reuteri ATG-F4 strain and a pharmaceutical excipient. The composition derived from the Lactobacillus reuteri ATG-F4 strain may be added to the pharmaceutical composition of the present invention in an amount of 0.001 to 100 weight%.

[0027] The above pharmaceutical composition may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, as well as external preparations, suppositories, and sterile injectable solutions, according to conventional methods. Carriers, excipients, and diluents that may be included in the above pharmaceutical composition may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulating, the preparation is made using diluents or excipients such as commonly used fillers, volume expanders, binders, humectants, disintegrants, and surfactants. Solid dosage forms for oral administration include tablets, pills, powders, granules, and capsules; these solid dosage forms are prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., with the extract of the present invention. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid dosage forms for oral administration include suspensions, liquids, emulsions, and syrups; in addition to commonly used simple diluents such as water and liquid paraffin, various excipients, for example, humectants, sweeteners, flavorings, and preservatives, may be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. As non-aqueous solvents and suspending agents, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As bases for suppositories, Witepsol, Macrogol, Tween 61, cacao oil, laurin oil, glycerogelatin, etc. may be used.

[0028] The dosage of the pharmaceutical composition of the present invention will vary depending on the age, gender, and weight of the subject to treatment, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration, and the judgment of the prescriber. The determination of the dosage based on these factors is within the level of a person skilled in the art, and generally, the dosage ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferred dosage is from 1 mg / kg / day to 500 mg / kg / day. Administration may be performed once a day or divided into several doses. The above dosage does not limit the scope of the present invention in any way.

[0029] The pharmaceutical composition of the present invention can be administered to mammals, such as rats, livestock, and humans, via various routes. Any mode of administration is expected, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater, or intracerebrovascular injection. Since the composition of the present invention has minimal toxicity and side effects, it is a drug that can be safely used for long-term preventive purposes. Effects of the invention

[0030] The present invention relates to Lactobacillus reuteri ATG-F4 ( Lactobacillus reuteri The present invention relates to a composition for combination administration comprising the strain ATG-F4 (accession number KCTC13717BP) and a chemical anticancer agent, wherein it is confirmed that tumor metastasis is significantly inhibited and the growth of the tumor itself is reduced through the combination administration of the strain and the anticancer agent.

[0031] In addition, in the generated tumors, the expression of cancer metastasis factors IL-6 receptor (Interleukin-6 receptor) and TGF-β (transforming growth factor-β) was significantly reduced, the apoptosis factors BAX (Bcl-2 associated X Protein) and Caspase fragments were increased, and the expression of tumor growth factors such as AKT (Akt8 virus transforming kinase target), ERK1 / 2 (Extracellular signal-regulated kinases 1 / 2), MEK1 (MAPK / ERK kinase 1), and COX-2 (Cyclooxygenase-2) was significantly reduced, suggesting that the combination of the Lactobacillus reuteri ATG-F4 strain and a chemical anticancer agent can be used as an excellent anticancer agent or metastasis inhibitor. Brief explanation of the drawing

[0032] Figure 1 shows the process of inducing tumors and metastasis in the experimental animals of Example 1. Figure 2 shows a photograph of a tumor taken from an experimental animal of Example 1. Figure 3 is a graph showing the average tumor weight results of the experimental animals in Example 1. Figure 4 is a Western blot result photograph confirming the tumor-associated protein factor expressed in the tumor of the experimental animal of Example 1. Figure 5 shows the results of confirming the distribution of the intestinal microbiota of the experimental animals of Example 1 and the butyric acid content in the cecum and blood. Specific details for implementing the invention

[0033] Preferred embodiments of the present invention will be described in detail below. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the content introduced herein is provided to fully convey the concept of the present invention to those skilled in the art, so that it may be thorough and complete.

[0034] <Example 1. Experimental Animals>

[0035] This animal experiment was approved as an appropriate procedure by the Institutional Animal Care and Use Committee (IACUC) of A2GEN Co., Ltd. (Approval No.: ATG-IACUCRDSP-220321), and ethical animal experiment guidance was provided. The animals used in this experiment were 5-week-old (Table 1) BALB / c nude mice purchased from Raon Bio (Seoul, Korea). HCT-116 cells (human colon cancer cell line, Korean Cell Bank) were cultured in DMEM (Dulbecco's modified Eagle's medium) supplemented with 10% FBS (fetal bovine serum). Mice were housed in a rearing room that maintained a constant indoor temperature of 23 ± 2℃, controlled temperature and humidity, and maintained a 12h:12h light cycle. A minimum acclimatization period of 7 days was provided to allow the mice to adapt to the new environment and feed, and there were no restrictions on water or rodent food (Purina) intake.

[0036] After the acclimatization period, 1.0 x 10 6 HCT-116 cells were administered subcutaneously to the flanks of mice, and tumors were grown for 10 days. The tumors were sliced ​​to a width x length x height of 1.5 mm and fixed to the mouse sigmoid colon using surgical sutures. During this process, the sigmoid colon was slightly scratched with a razor blade to promote tumor engraftment. After transplanting the tumors into the sigmoid colon, a recovery period of 7 days was observed. Subsequently, FOLFOX (Oxaliplatin 6 mg / kg, 5-Fluorouracil 50 mg / kg, Leucovorin (calcium folinate) 90 mg / kg) was administered intraperitoneally, and 4.0 x 10 10ATG-F4 in CFU was administered orally daily. The anticancer agent FOLFOX was administered 10 times over a total of 20 days, whenever the animal's body weight recovered after the anticancer agent administration. This process is shown in Figure 1 and Table 1.

[0037] Group n number Normal Control (NC) 10 Cancer 4 Cancer + FOLFOX (Cancer Fx) 11 Cancer + FOLFOX + ATG-F4 (Cancer Fx ATG-F4) 11

[0038] <Example 2. Confirmation of the condition of mice administered ATG-F4 strain and anticancer drug>

[0039] The mice were autopsied by CO2 euthanasia on the last day of the experiment.

[0040] The condition photos of each tumor are as shown in Fig. 2, and the average weight of the tumors is shown in Fig. 3.

[0041] Looking at Figures 2 and 3, it is confirmed that the tumors of the mice in the group that were not administered anticancer drugs grew very large, both metastatic tumors and colon tumors. In mice administered only chemotherapy drugs, the volume of the tumors decreased for both metastatic tumors and colon tumors, but when comparing this to the group administered chemotherapy drugs and the ATG-F4 strain in combination in Figure 3, it is confirmed that the tumor weight decreased by about half.

[0042] <Example 3. Confirmation of Anti-tumor / Anti-metastasis Related Protein Expression - Western Blot Analysis>

[0043] Mouse colon tumor tissue was placed in RIPA buffer (0.5 M Tris-HCl, pH 7.4, 1.5 M NaCl, 2.5% deoxycholic acid, 10% NP-40) containing a protease inhibitor cocktail (Millipore, USA) and chopped with scissors. Subsequently, the tissue was separated in a centrifuge at 14,000 rpm at 4°C for 10 minutes, and the supernatant was collected. The protein concentration in the supernatant was measured using a BCA assay (Thermo Fisher, USA). The protein extract (40 μg) was separated using an 8% or 10% polyacrylamide mini-gel and transferred to a PVDF membrane (Bio-Rad, USA). The transcribed PVDF membranes were incubated overnight at 4°C with SuperBlock (PBS) Blocking buffer (pH 7.4) containing Kathon™ Antimicrobial Agent and primary antibodies related to tumors and metastases, respectively. The assay parameters were apoptosis factors (BAX, Caspase), tumor growth factors (AKT, ERK1 / 2, MEK1, COX-2), and metastasis-related factors (IL-6, TGF-β). The PVDF membranes were washed four times with 0.1% Tween TBS and then placed in 0.1% Tween TBS buffer containing 3% BSA (Bovogen, USA) and incubated at room temperature for one hour with goat anti-rabbit IgG HRP conjugated secondary antibody (Bio-Rad, USA). After thorough washing with 0.1% Tween TBS, the immunostained bands were verified using ECL (Bio-Rad, USA). Target proteins were identified using the ChemiDoc™ Imaging System (Bio-Rad, USA), and band intensity was quantified using Image Lab™ software (Bio-Rad, USA).

[0044] The results of this are shown in Figure 4. As a result, in the group administered with the anticancer agent FOLFOX and the ATG-F4 strain, the protein expression levels of apoptosis-related BAX (Figure 4a) and Caspase-3 fragment (Figure 4b) increased, while tumor growth / proliferation-related AKT (Figure 4c), ERK1 / 2 (Figure 4d), MEK1 (Figure 4e), and COX-2 (Figure 4f) decreased, and the expression of metastasis-related IL-6 receptor (Figure 4g) and TGF-β (Figure 4h) was significantly reduced.

[0045] Accordingly, it can be confirmed that the efficacy of the anticancer drug FOLFOX is very effectively boosted through the combination administration of ATG-F4, which produces effects that were not well observed with the anticancer drug FOLFOX alone, through the processes of apoptosis, inhibition of tumor growth and proliferation, and inhibition of tumor metastasis.

[0046] <Example 4. Confirmation of efficacy in inducing changes in gut microbiota and increase in butyric acid>

[0047] Amplicons of the V3-V4 region of 16S rRNA were obtained via PCR from metagenomic DNA extracted from appendiceal samples using the QIAamp PowerFecal Pro DNA kit (Qiagen, Germany), and sequencing was performed using the Miseq platform, a next-generation sequencing instrument. Changes in the gut microbiota caused by LT-002 were analyzed using the Qiime2 data analysis pipeline and the SILVA 138 database.

[0048] Butyric acid analysis in serum and cecum was performed using the SCIEX Triple Quad™ 4500 LC-MS / MS System. The process involved standard preparation, sample pretreatment, derivatization, and LC-MS / MS analysis. Serum and cecum samples underwent deproteinization and filtration using 1% formic acid in methanol, while an additional derivatization step was performed for butyric acid analysis. 0.1% formic acid in water / acetonitrile was used as the mobile phase for the LC-MS / MS analysis, and the results were analyzed using SCIEX OS 2.1.0 and Analyst 1.7.1 software.

[0049] Through this process, the gut microbiome of each mouse individual euthanized in Example 2 was identified. The results are shown in the following Figure 5a via the LDA score. This LDA score indicates how characteristically each bacterium is distributed in the experimental mouse group; a higher LDA score indicates that the corresponding bacterium is present in greater quantities in the experimental group of the present invention.

[0050] In other words, looking at Figure 5a, it was confirmed that compared to NC, there was a slight effect on the changes in the gut microbiome upon cancer cell transplantation, and a significant change in the gut microbiome was observed in the group administered the anticancer drug FOLFOX alone, and in particular, the combined administration of the anticancer drug and the ATG-F4 strain showed a very different pattern.

[0051] When classifying the increase or decrease by each bacterial colony, it was shown that there was a distinct differentiation between the group administered the anticancer drug alone and the group administered the anticancer drug in combination with the ATG-F4 strain. Among these, Butyricicoccus and others were found to increase in the group administered the anticancer drug in combination with the ATG-F4 strain. Meanwhile, Escherichia and Shigella, which are well-known harmful bacteria, increased due to the administration of the anticancer drug, but decreased due to the combined administration of the anticancer drug and ATG-F4, which indicated a return to the state prior to the administration of the anticancer drug and the occurrence of cancer.

[0052] Similarly, in the group administered the anticancer drug alone, butyric acid (BA) production in the cecum and blood was reduced, but through the increase of Butyricicoccus, it can be seen that in the group administered the anticancer drug and the ATG-F4 strain in combination, as shown in Figures 5b and 5c, butyric acid production was maintained in a state where no change in the intestinal microbiome occurred (NC).

[0053] Through the above results, it is proven that the combination administration composition of the Lactobacillus reuteri ATG-F4 strain of the present invention and an anticancer agent is an excellent composition for anticancer or tumor metastasis inhibition.

Claims

Claim 1 Lactobacillus reuteri ATG-F4( Lactobacillus reuteri A composition for antitumor or tumor metastasis inhibition characterized by containing the strain ATG-F4 (accession number KCTC13717BP) as an active ingredient. Claim 2 A composition for inhibiting tumor or tumor metastasis according to claim 1, characterized in that the strain comprises one or more selected from the group consisting of the cell, the culture of the strain, the extract, the concentrate, and the dried product of the culture. Claim 3 The composition for antitumor or tumor metastasis inhibition according to claim 1, characterized in that the composition increases the expression of BAX (Bcl-2 associated X Protein) and Caspase-3 fragment proteins, or inhibits the protein expression of IL-6 receptor (Interluekin-6 receptor), TGF-β (transforming growth factor-β), AKT (Akt8 virus transforming kinase target), ERK1 / 2 (Extracellular signal-regulated kinases 1 / 2), MEK1 (MAPK / ERK kinase 1), and COX-2 (Cyclooxygenase-2). Claim 4 A composition for inhibiting tumor or tumor metastasis according to claim 1, characterized in that a chemical anticancer agent is administered in combination with the composition. Claim 5 In claim 1, the chemical anticancer agent is oxaliplatin, cisplatin, cyclophosphamide, ifosfide, bendamustine, melphalan, carboplatin, busulfan, dacarbazine, temozolomide, fluorouracil, capecitabine, doxifluridine, tegafur, cytarabine, azacitidine, decitamine, enositabine, methotrexate, pemetrexid, pralatrexate, cladribine, cloparabine, fludarabine, mercaptopurine, irinotecan, doxorubicin, daunorubicin, epirubicin, idarubicin, mitoxantrone, etoposide, topotecan, cabazitaxel, paclitaxel, docetaxel, vinblastine, vincristine, vinorelbine, bleomycin, hydroxyurea, and mitomycin C. A composition for antitumor or tumor metastasis inhibition characterized by selecting one or more types from the group formed. Claim 6 An anticancer or anti-metastatic pharmaceutical composition containing the composition of claim 1 and pharmaceutical excipients.

Images