JPWO2010023716A1 - Microbial culture having anticancer activity and product using the microorganism culture - Google Patents

Abstract

[PROBLEMS] To suppress cancer cell proliferation, to suppress cancer cell invasion, to enhance cancer healing power, and to be highly versatile, and to further suppress cancer cell proliferation, suppress cancer cell invasion, enhance cancer healing power, etc. Provided are pharmaceuticals that have anticancer effects and have no side effects, and raw materials for quasi-drugs for prevention and improvement. A microorganism [Lactobacillus sp. SIID1719-6b (original accession number FERM AP-20062, new accession number FERM ABP-10299)] isolated from kefir grains, or a microorganism containing the microorganism A microorganism culture obtained by culturing a group with a combination of green tea and a tea other than green tea and / or an active ingredient thereof, and having an anticancer activity. [Selection figure] None

Description

  The present invention provides anticancer agents, immunostimulants, therapeutic agents, and other drugs, as well as raw materials for quasi-drugs for prevention and improvement, or by adding to these products or raw materials to suppress cancer cell proliferation, cancer cell infiltration The present invention relates to a microorganism culture having anticancer effects such as suppression and enhancement of cancer healing ability. More specifically, the present invention relates to a microorganism culture obtained by culturing microorganisms in combination with green tea and tea other than green tea and / or an active ingredient thereof. Furthermore, the present invention relates to pharmaceuticals such as anticancer agents, immunostimulants and therapeutic agents using them and quasi-drugs for prevention and improvement.

As a cause of death in Japan, malignant neoplasms (cancers) continue to rise consistently, becoming the first cause of death after 1981, accounting for 30.1% of all deaths in 2005. Approximately 1 in 3 deaths will be due to malignant neoplasm (cancer). Therefore, suppression and prevention of cancer are important issues in today's medicine.

In animal pets, cancer is the leading cause of death for dogs and cats.

Examples of cancer treatment methods include surgery and radiation therapy, and their progress is remarkable. However, there are problems in the case of recurrence and metastasis of cancer in surgery, and in radiotherapy, there are cases where it is ineffective due to differences in sensitivity depending on the type of cancer. Therefore, it is effective for local cancer such as the primary lesion, but it is almost impossible to completely remove it when the cancer has recurred or metastasized.

In addition, as a cancer treatment method, one of the three major cancer treatment methods including the above-described surgery and radiation treatment includes chemotherapy (anticancer drug treatment). The types of anticancer agents used in this case include “cytotoxic anticancer agents” and “molecular target therapeutic agents”, of which the former “cytotoxic anticancer agents” include antimetabolites, alkylating agents, and anticancer antibiotics. There are substances, microtubule inhibitors.

However, in the case of a “cytotoxic anticancer agent” as a systemic therapy, the effect and adverse drug reaction (side effects) may be half or the adverse drug reaction may be greater than the effect. Major adverse drug reactions include symptoms such as nausea, vomiting, numbness, hair loss, leukopenia and fever, thrombocytopenia and bleeding, liver dysfunction, and renal dysfunction. Efforts have been made to reduce adverse drug reactions and molecular targeted therapeutic drugs have been developed. However, drug adverse reactions have not been completely eliminated.

Examples of treatment methods other than the above three major cancer treatment methods include immune cell therapy and gene therapy. However, it is not only disadvantageous in terms of cost, such as the lack of insurance coverage, but also in the clinical trial stage.

On the other hand, anticancer supplements include agaricus, mesimacob, activated polysaccharide-related compound (AHCC), maitake extract, and yamabushitake as mushroom supplements, and fucoidan and arabinoxylan as plant supplements. However, some have problems such as damage due to large intake, allergic diseases, and inclusion of carcinogens.

  Conventionally, anticancer agents have many side effects and precautions regarding handling. Therefore, cancer cell proliferation inhibitory action, cancer cell invasion inhibitory action, cancer healing power enhancing action is strong, and versatility is high, furthermore cancer cell growth inhibition, cancer cell invasion inhibition, cancer healing power There has been a demand for development of raw materials for quasi-drugs for prevention and improvement, which are safe and have no side effects and have anticancer effects such as potentiation.

Among the techniques using microbial cultures, anticancer immunotherapeutic agents (eg, patents) containing enterococci faecalis enterococcus faecalis and docosahexaenoic acid as active ingredients as therapeutic agents for cancer using natural lactic acid bacteria Reference 1), an immunoreinforcing agent composition of poly-γ-glutamic acid and an antigenic substance, such as peptides, polypeptides, lactic acid bacteria that express the polypeptides, antigen proteins, lactic acid bacteria that express the antigen proteins, etc. A composition for vaccines for preventing or treating (for example, refer to Patent Document 2), fermentation to selectively add lactic acid bacteria and at least one yeast in order to suppress cancer and reduce the risk of infectious diseases Therapeutic composition containing soy extract (see, for example, Patent Document 3), apoptosis induction and inflammation in autoimmune diseases such as cancer Lactic acid bacteria highly enriched with arginine deiminase (eg, see Patent Document 4), a preventive or therapeutic agent for tumors including cancer, at least one selected from pearl husk, thin skin and astringent skin, A food or pharmaceutical composition obtained by fermenting with at least one kind of microorganism selected from lactic acid bacteria and yeast (for example, see Patent Document 5) has been proposed.

Japanese Patent Laid-Open No. 8-27010

JP 2006-232799 A

JP 2003-26582 A

Special Table 2007-528848

International Publication No. 02/072123 Pamphlet

Among the techniques using natural tea, polyphenol derivatives extracted from tea leaves (for example, see Patent Document 6) and oolong tea extract are effective as pharmaceuticals and health supplements that induce apoptosis in cancer cells. A cancer cell metastasis inhibitor (see, for example, Patent Document 7) characterized by being an ingredient, purified from Rubois tea, which has a cancer cell proliferation inhibitory action and a cancer cell invasion inhibitory action containing aspartin as an active ingredient An anti-cancer agent (see, for example, Patent Document 8) characterized by being a substance has been proposed.

JP-A-2005-75790

JP 2001-39881 A

JP 2007-197409 A

On the other hand, the traditional fermented milk kefir (Kefir) of lactic acid bacteria and yeast, which is considered the birthplace of the Caucasus region, is produced by culturing kefir grains, which are the starter, in milk.
The present inventor has found that when this kefir grain, which is a natural product, is used for beverages, foods, cosmetics, feeds, etc., it is safe for humans and animals, and further, earnestly researches various utilization methods. I came. And, a microorganism [Lactobacillus sp. SIID1719-6b] separated from kefir grains, or a microorganism culture obtained by culturing a group of microorganisms containing the microorganism, and a product using them, It came to develop (for example, refer patent document 9). Further, a microorganism [Lactobacillus sp. SIID1719-6b] isolated from kefir grains, or a microorganism group containing the microorganism as a microorganism culture obtained from a medium mainly composed of green tea and black tea, and The product used was proposed and developed (for example, refer to Patent Document 10). Further, a microorganism culture obtained by using a microorganism [Lactobacillus sp. SIID1719-6b] isolated from kefir grains, or a group of microorganisms containing the microorganism as a main component of green tea, black tea and mugwort extract Proposed and developed products and products using the same (for example, see Patent Document 11). Further, a skin obtained by culturing a microorganism [Lactobacillus sp. SIID1719-6b] isolated from kefir grains or a group of microorganisms containing the microorganism in a green tea culture solution, Additives to drugs (ointments), dermatological drugs, cosmetics, orally administered drugs, and quasi-drugs for prevention and improvement have the effect of suppressing the onset of dermatitis and promoting healing of skin wounds Microbial cultures and products using them have been proposed and developed (for example, see Patent Document 12).

EP 1607476

JP 2006-271219 A

JP 2006-271220 A

JP 2007-137864 A

  As a result of further research, the inventor is a natural product that has high safety if it is a microbial culture obtained by culturing microorganisms in combination with green tea and tea other than green tea and / or its active ingredients. , If they are used, anticancer agents having anticancer activity such as cancer cell growth inhibition, cancer cell invasion inhibition, enhancement of cancer healing power, immunopotentiators, therapeutic drugs and other quasi-drugs for prevention and improvement It was found that a raw material of In addition, we found that the effect differs depending on the type of cancer by changing the combination of green tea and tea other than green tea.

Based on these findings, the present inventor obtained a microorganism [Lactobacillus sp. SIID1719-6b] isolated from kefir grains, or a group of microorganisms containing the microorganism as a tea other than green tea and green tea and / or Or a microbial culture obtained by microbial culture in combination with an active ingredient thereof can be used as a raw material for pharmaceuticals such as anticancer agents, immunostimulants, therapeutic agents, and quasi drugs for prevention and improvement, The present inventors have found that by adding the microbial culture to these products or raw materials, they have anticancer effects such as suppression of cancer cell proliferation, suppression of cancer cell infiltration, and enhancement of cancer healing power. They also found that changing the combination of green tea and non-green tea has different effects depending on the type of cancer. And this inventor added cancer culture growth, cancer by adding the said microbial culture to pharmaceuticals, such as an anticancer agent, an immunostimulant, and a therapeutic agent using them, and a quasi-drug for prevention and improvement. This has led to the development of products with anticancer effects such as suppression of cell invasion and enhancement of cancer healing power.

The present invention is an invention having the following configuration. That is,
(1) A microorganism isolated from Kefir grains [Lactobacillus sp. SIID1719-6b (original accession number FERM AP-20062, new accession number FERM ABP-10299)] or the microorganism [Lactobacillus sp. (Lactobacillus sp.) Microbial culture containing SIID1719-6b (original deposit number FERM AP-20062, new deposit number FERM ABP-10299)] in combination with green tea and tea other than green tea and / or active ingredients thereof A microorganism culture obtained as described above, which has an anticancer activity.
(2) The microorganism culture according to (1) above, wherein the anti-cancer effect can be enhanced by combining black tea as green tea and tea other than green tea.
(3) The above-mentioned (1) characterized in that it is possible to expand the types of cancer to be cured and enhance the anticancer effect by combining black tea and Tochu tea as tea other than green tea and green tea. ) Of the microorganism culture described.
(4) The microorganism culture according to any one of (1) to (3) above, wherein the anticancer effect is an effect of suppressing cancer cell proliferation, cancer cell invasion and the like and an effect of enhancing cancer healing power.
(5) The above-mentioned cancers are neuroblastoma, breast cancer, uterine cancer, hepatocellular carcinoma, renal cell carcinoma, pancreatic cancer, bladder, renal pelvis, ureteral cancer, colon cancer, skin cancer (malignant melanoma, basal cell carcinoma, spiny) Cell cancer), pharyngeal cancer, laryngeal cancer, thyroid cancer (papillary cancer, follicular cancer, medullary cancer), lung cancer, esophageal cancer, stomach cancer, biliary tract cancer, duodenal papilla cancer, prostate cancer, ovarian cancer and brain tumor (medulloblastoma) , Germ cell tumor), malignant mesothelioma, thymoma, testicular tumor, uterine sarcoma, bone and soft tissue tumor, leukemia, malignant lymphoma, the microorganism culture according to any one of (1) to (3) above is there.
(6) The microorganism culture obtained by removing the cells from the microorganism culture described in (1) also has the same effects and functions as those described above, that is, the anticancer action and functions.
(7) A pharmaceutical product such as an anticancer agent having an anticancer activity, an immunostimulant, a therapeutic agent, and a quasi-drug for prevention and improvement, characterized in that the microorganism culture according to (1) is added It is.

  According to invention of Claim 1, microorganisms [Lactobacillus sp. SIID1719-6b] separated from kefir (Kefir) grains, or a group of microorganisms containing the microorganisms, green tea and tea other than green tea and It is a microorganism culture obtained by culturing microorganisms in combination with an active ingredient thereof and / or has an effect of newly providing a microorganism culture having an anticancer activity.

According to the invention described in claim 2, it is possible to enhance the anti-cancer effect by combining black tea as green tea and tea other than green tea. There is an effect that can be newly provided.

  According to the invention described in claim 3, by combining black tea and Tochu tea as tea other than green tea and green tea, it becomes possible to expand the types of cancer to be cured and enhance the anticancer effect, There is an effect that a microbial culture can be newly provided according to the type and price.

  According to the invention described in claim 4, there is an effect that it is possible to newly provide a microorganism culture having an anti-cancer effect such as an effect of suppressing cancer cell proliferation, cancer cell invasion and the like and an effect of enhancing cancer healing power.

According to the invention described in claim 5, there is an effect that a microorganism culture having an anticancer action can be newly provided for cancers and tumors related to body surfaces and organs.

  According to invention of Claim 6, it is a microorganism culture which removed the microbial cell from the culture of Claim 1, Comprising: There exists an effect which can newly provide the microorganism culture which has anticancer action.

According to claim 7, the microbial culture is safe without toxicity and side effects, and the antimicrobial agent having an anticancer activity, an immunostimulant, a therapeutic agent, and the like, to which the microbial culture is added, and prevention There is an effect that a quasi-drug for improvement can be newly provided.

A preferred embodiment of the present invention will be described in detail.
Microorganisms isolated from kefir grains [Lactobacillus
sp.) SIID1719-6b] was subjected to the following evaluation test to determine whether the microorganism culture obtained by culturing microorganisms with a combination of green tea and tea other than green tea has cancer cell proliferation inhibitory action and function. As a result, it was confirmed that the microbial culture obtained by changing the combination of green tea and tea other than green tea and cultivating the microorganism had cancer cell growth inhibitory action and function. Furthermore, in the present invention, microorganisms [Lactobacillus sp. SIID1719-6b] and microorganisms that have been confirmed to be safe and exhibit similar actions, teas other than green tea and green tea, and / or their effectiveness Any of them can be used as long as they are prepared in combination with components.
In addition, microorganisms [Lactobacillus sp. SIID1719-6b (FERM
AP-20062)] has been deposited at the National Institute of Advanced Industrial Science and Technology, Patent Biological Depositary Center, and has been internationally deposited at the center as FERM ABP-10299.

As a representative example, a microorganism [Lactobacillus sp. SIID1719-6b] is mixed with two kinds of green tea and black tea in a ratio of 5: 5, and green tea and black tea in a ratio of 3: 4: 3. Two types of three kinds of mixed teas that combined with Tochu tea were cultivated with microorganisms, and cancer cell growth inhibition evaluation tests were carried out as follows using the obtained two types of microbial cultures.

Human breast cancer cell HS-538T, human uterine cancer cell HeLa, human hepatocellular carcinoma cell HepG2, human renal cell carcinoma cell Caki-1 and KU19-20, human pancreatic cancer cell MiaPaCa-2, human bladder cancer cell RYOJI-XA, hamster pancreas A total of 9 types of cancer cells including β-cell HIT-T15 and adenovirus-transformed human embryonic kidney cells HEK293 were treated with 10 mM.
10% FBS, penicillin and streptomycin were added to a DMEM / F12 medium (Sigma D6421) containing HEPES, cultured at 37 ° C. in a 5% CO ₂ environment, cells in the growth phase were treated with trypsin, suspended in the medium, and coated with collagen. A 96-well plate was plated to be about 20% confluent.

After culturing for 15 hours, the medium was aspirated, and the microorganisms [Lactobacillus sp. SIID1719-6b] were adjusted so that the respective concentrations were 0%, 1%, 2%, 4%, 8% and 16%. Serum-added 10 mM with a 48-hour culture (hereinafter referred to as B2 solution (a)) of two types of mixed tea combining green tea and black tea in the above proportions
HEPES-containing DMEM / F12 medium was added at 100 μL per well and cultured at 37 ° C. for 40 hours in a 5% CO ₂ environment. Each treatment was conducted in 4 wells.

In the same manner as described above, after culturing for 15 hours, the medium is aspirated, and the microorganisms [Lactobacillus genus ((Lactobacillus sp.)] Are adjusted so that the respective concentrations become 0%, 1%, 2%, 4%, 8% and 16%. Lactobacillus sp.) SIID 1719-6b] and 48 mM culture (hereinafter referred to as B11 solution (b)) of 3 kinds of mixed teas combining green tea, black tea and Tochu tea in the above proportions and 10 mM serum added
HEPES-containing DMEM / F12 medium was added at 100 μL per well and cultured at 37 ° C. for 40 hours in a 5% CO ₂ environment. As in the above case, each treatment was conducted in 4 wells.

After the culture, the medium was aspirated, and serum-added 10 mM supplemented with 10% AlarmBlue (BIOSOURCE) was used to quantify the growth rate of cancer cells.
HEPES-containing DMEM / F12 medium (100 μL) was added and cultured at 37 ° C. for 4 hours in a 5% CO ₂ environment. The absorbance (570 nm, 595 nm) of the medium was measured, and the AlarmBlue reduction rate was calculated by the following calculation formula (1). Growth inhibition for each cancer cell was determined as a relative value when the value of 0% of the culture supernatant was 100%.
The results are shown in FIGS. 1 to 9 for graphs of relative values for each cancer cell. (The vertical axis represents the cancer cell growth rate, and the horizontal axis represents the culture supernatant concentration.)

Human breast cancer cell HS-538T, human uterine cancer cell HeLa, human hepatocellular carcinoma cell HepG2, human renal cell carcinoma cell Caki-1 and KU19-20, human pancreatic cancer cell MiaPaCa-2, human bladder cancer cell RYOJI-XA, hamster pancreas The hamster pancreatic β cell HIT-T15 was not able to obtain a significant difference compared with the total of 9 types of cancer cells, β cell HIT-T15 and human embryonic kidney cell HEK293 transformed with adenovirus, but the remaining 8 For various types of cancer cells, suppression of cancer cell proliferation was observed depending on the concentration of the culture supernatant.

Moreover, it was recognized that the B11 solution (b) was significantly higher than the B2 solution (a) with respect to the 8 types of cancer cells.

Furthermore, the growth inhibitory effect when the B2 solution (a) or the B11 solution (b) is added at a 10-fold dilution is obtained for human uterine cancer cells HeLa, human pancreatic cancer cells MiaPaCa-2, and human bladder cancer cells RYOJI-XA. The growth inhibition state on the fifth day was observed with a phase contrast microscope. The photograph which image | photographed the growth suppression state of the said 3 types of cancer cells is shown in FIGS. The left side of each figure shows the case where the B2 solution (a) was added at 10-fold dilution, and the right side of each figure shows the case where the B11 solution (b) was added at 10-fold dilution. When B2 solution (a) or B11 solution (b) was not added, the cancer cells spread and grew throughout the dish.

From these, microorganisms isolated from Kefir grains [Lactobacillus
sp.) SIID1719-6b], or the microorganism group containing the microorganisms as a microorganism culture according to the present invention obtained by culturing microorganisms with a combination of green tea and tea other than green tea and / or active ingredients thereof. It is suggested that cancer cell growth has an inhibitory effect, and that it is possible to enhance the cell growth inhibitory effect for each type of cancer by changing the combination of green tea and tea other than green tea. It was.

  A microorganism [Lactobacillus sp. SIID1719-6b] isolated from kefir grains, or a microorganism group containing the microorganism, in combination with green tea and tea other than green tea and / or active ingredients thereof. Microbial cultures obtained by culturing microorganisms can be expected to have anticancer effects such as suppression of various types of cancer cell growth, suppression of cancer cell infiltration, and enhancement of cancer healing power.

The microorganism culture can be used as a raw material for pharmaceuticals such as anticancer agents, immunostimulants, therapeutic agents, and quasi-drugs for prevention and improvement, and the microorganism culture is added to these products or raw materials. By doing so, anticancer effects such as suppression of cancer cell proliferation, suppression of cancer cell infiltration, and enhancement of cancer healing ability can be expected.

In order to re-identify the species of SIID1719-6b after depositing at the National Institute of Advanced Industrial Science and Technology Patent Biological Depositary Center, using the nucleotide sequence described in European Patent No. 1607476 (Patent Document 9), 16S obtained by homology search against the bacterial reference strain database
A homology search of species considered to be closely related to the specimen was performed from the base sequence of rDNA, and the top 5 strains were determined.
The homology search results are shown in Table 1.

Based on the information obtained so far, a molecular phylogenetic tree was constructed using a 16S rDNA base sequence and a 16S rDNA base sequence of a bacterial group considered to be closely related thereto. Note that the neighbor linking method was used to estimate the molecular phylogenetic tree, and a bootstrap indicating the validity of the tree type was generated 1000 times. MEGA for analysis
ver. 3.1 was used.
The molecular phylogenetic analysis results are shown in FIG.

Lactobacillus satsumensis, which is the closest relative to the base sequence analysis of 16S rDNA, was subjected to a DNA-DNA hybridization test to determine species differences.
As a method, anaerobic culture was performed with a culture medium MRS Broth (Oxoid, Hampshire, UK) at a culture temperature of 30 ° C. and a culture time of 48 hours. From the cultured cells of each specimen, Kawamura et al. Bacteriological Society, August 2000,
55, No. 3, p. 545-584), DNA was extracted and purified. Main methods of DNA-DNA hybridization test are Kawamura et al.'S method and book (Kenichiro Suzuki, Akira Hiraishi, Akira Yokota, “Microbial Classification / Identification Experiment”, Molecular Genetics / Molecular Biology) And Springer Japan, September 16, 2001, p.34-47) using the microplate method. Fluorescence plate reader (Genius,
TECAN (Wako Pure Chemical Industries, Ltd., Osaka)) was used.

SIID1719-6b and Lactobacillus
The average of homologous values by three tests of satsumensis showed a value of 70% or more. At present, bacterial species are defined as strains showing homology values of 70% or more as the same species as a result of comparison of DNA-DNA homology values. From this, it was judged that SIID1719-6b and Lactobacillus satsumensis were the same kind.
Therefore, SIID 1719-6b is a Lactobacillus
It was identified as satsumensis.

  Table 2 shows the base sequence of the 16S rDNA base sequence of SIID1719-6b.

It is a graph which shows the relative value of the cancer cell proliferation inhibitory effect with respect to human breast cancer cell HS-538T. It is a graph which shows the relative value of the cancer cell growth inhibitory effect with respect to the human uterine cancer cell HeLa. It is a graph which shows the relative value of the cancer cell proliferation inhibitory effect with respect to human hepatocellular carcinoma cell HepG2. It is a graph which shows the relative value of the cancer cell proliferation inhibitory effect with respect to human renal cell carcinoma cell Caki-1. It is a graph which shows the relative value of the cancer cell proliferation inhibitory effect with respect to human pancreatic cancer cell MiaPaCa-2. It is a graph which shows the relative value of the cancer cell growth inhibitory effect with respect to the human bladder cancer cell RYOJI-XA. It is a graph which shows the relative value of the cancer cell growth inhibitory effect with respect to hamster pancreatic beta cell HIT-T15. It is a graph which shows the relative value of the cancer cell proliferation inhibitory effect with respect to human renal cell carcinoma cell KU19-20. It is a graph which shows the relative value of the cancer cell growth inhibitory effect with respect to the human embryonic kidney cell HEK293 transformed with adenovirus. In the photograph which shows the growth suppression state of the 5th day with respect to human uterine cancer cell HeLa, the case where the left is B2 solution (a) and the right side is the case of B11 solution (b) is shown. It is the photograph which shows the growth suppression state of the 5th day with respect to human pancreatic cancer cell MiaPaCa-2, The case where the left side is B2 solution (a), and the case where the right side is B11 solution (b) are shown. The photograph which shows the growth suppression state of the 5th day with respect to the human bladder cancer cell RYOJI-XA, the case where the left side is B2 solution (a), and the case where the right side is B11 solution (b) are shown. It is a phylogenetic tree showing the results of molecular phylogenetic analysis.

Claims (7)

Microorganisms isolated from Kefir grains [Lactobacillus
sp.) SIID1719-6b (original accession number FERM AP-20062, new accession number FERM ABP-10299)] or the microorganism [Lactobacillus sp.) SIID1719-6b (original accession number FERM AP-20062, new A microorganism culture obtained by culturing a microorganism group including the accession number FERM ABP-10299)) with a combination of green tea and tea other than green tea and / or an active ingredient thereof, and having anticancer activity A microbial culture characterized by 2. The microorganism culture according to claim 1, wherein the anti-cancer effect can be enhanced by combining black tea as green tea and tea other than green tea. The microorganism according to claim 1, characterized in that, by combining black tea and Tochu tea as tea other than green tea and green tea, it is possible to expand the types of cancer to be cured and enhance the anticancer effect. Culture. The microorganism culture according to any one of claims 1 to 3, wherein the anti-cancer effect is an effect of suppressing cancer cell proliferation, cancer cell infiltration and the like, and an enhancement effect of cancer healing power. The above cancers are neuroblastoma, breast cancer, uterine cancer, hepatocellular carcinoma, renal cell carcinoma, pancreatic cancer, bladder, renal pelvis, ureteral cancer, colon cancer, skin cancer (malignant melanoma, basal cell carcinoma, squamous cell carcinoma). Pharyngeal cancer, laryngeal cancer, thyroid cancer (papillary cancer, follicular cancer, medullary cancer), lung cancer, esophageal cancer, stomach cancer, biliary tract cancer, duodenal papilla cancer, prostate cancer, ovarian cancer and brain tumor (medulloblastoma, germ cell) The microbial culture according to any one of claims 1 to 4, which comprises malignant mesothelioma, thymoma, testicular tumor, uterine sarcoma, bone soft tissue tumor, leukemia, and malignant lymphoma. A microorganism culture obtained by removing cells from the microorganism culture according to claim 1. A pharmaceutical product such as an anticancer agent having an anticancer activity, an immunostimulatory agent, a therapeutic agent, and a quasi-drug for prevention and improvement, characterized in that the microorganism culture according to claim 1 is added.