WO2000033069A1 - Substance contenant un extrait d'hypes du champignon shiitake destinee a cribler l'activite lak, et procede mettant en oeuvre ladite substance pour cribler l'activite lak - Google Patents

Substance contenant un extrait d'hypes du champignon shiitake destinee a cribler l'activite lak, et procede mettant en oeuvre ladite substance pour cribler l'activite lak Download PDF

Info

Publication number
WO2000033069A1
WO2000033069A1 PCT/JP1999/006615 JP9906615W WO0033069A1 WO 2000033069 A1 WO2000033069 A1 WO 2000033069A1 JP 9906615 W JP9906615 W JP 9906615W WO 0033069 A1 WO0033069 A1 WO 0033069A1
Authority
WO
WIPO (PCT)
Prior art keywords
screening
lak
cells
activity
substance
Prior art date
Application number
PCT/JP1999/006615
Other languages
English (en)
Japanese (ja)
Inventor
Kenji Asano
Yukiko Matsuda
Yutaka Tajima
Original Assignee
Kobayashi Pharmaceutical Co., Ltd.
Nagaoka, Hitoshi
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kobayashi Pharmaceutical Co., Ltd., Nagaoka, Hitoshi filed Critical Kobayashi Pharmaceutical Co., Ltd.
Priority to CA002352614A priority Critical patent/CA2352614C/fr
Priority to GB0112999A priority patent/GB2360090B/en
Priority to KR1020017006475A priority patent/KR20010089498A/ko
Publication of WO2000033069A1 publication Critical patent/WO2000033069A1/fr
Priority to HK02101648.2A priority patent/HK1041518B/zh
Priority to HK02104333.6A priority patent/HK1042745A1/zh

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/07—Basidiomycota, e.g. Cryptococcus
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5011—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing antineoplastic activity
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2500/00—Screening for compounds of potential therapeutic value

Definitions

  • the present invention relates to the field of tumor immunology.
  • the present invention relates to substances and methods for screening for anti-tumor and immunotherapeutic agents with Z or anti-cancer activity. More specifically, the present invention relates to a screening substance for determining whether or not the effect of enhancing the activity of LAK cells (Ly immediate hokine Activated Killer Cell) in vivo can be obtained in vitro. And its method.
  • LAK cells Ly immediate hokine Activated Killer Cell
  • tumor cells have tumor antigens.
  • Tumor specific antigens TSA: Tumor Specific Antigen
  • TAA Tumor Specific Antigen
  • TAA tumor associated antigens
  • Such a tumor antigen is newly expressed due to a gene mutation caused by canceration of normal cells, or is expressed by a change in the regulation of the expression accompanying the mutation.
  • Immunotherapy is the most common treatment for tumor cells with abnormal antigenicity, such as drugs that immunize a subject with tumor antigens or enhance the immune function of the subject. .
  • NK cells are non-T non-B cytotoxic lymphoid cells that exist in normal individuals, and do not express MHC class I molecules without being restricted by MHC antigens against tumor cells, virus-infected cells, etc. It is known that it shows a damaging activity on cells whose expression is reduced. However, it has now become apparent that there are tumor cells that cannot be killed by NK cells.
  • S. Rosenberg of the US National Cancer Institute (XCI) states that when peripheral lymphocytes are cultured with Inuichi Leukin 2 (IL-2), they can target a wide range of target cancer cells, including autologous cancers.
  • LAK cell lymphokine-activated killer cell
  • LAK adoptive immunotherapy using IL-2 when LAK adoptive immunotherapy using IL-2 is performed, symptoms such as general malaise, chills, fever, low albumin, anemia, and eosinophilia are produced as side effects. 2 is known to be stronger than when used alone.
  • the occurrence of several important side effects has been attributed to the possibility that LAK cells have cytotoxic activity against normal cells. It has been reported that such hematopoietic stem cell damage by LAK cells causes anemia and thrombocytopenia, as well as in vitro damage to lymphocytes and macrophage ⁇ endothelial cells.
  • IL-12 has poor absorption by oral administration, and at present, injection is the mainstream for direct administration.
  • bacteria or foods have an anticancer effect. Since these bacteria or foods having an anticancer effect have few side effects and are safe, search for anticancer substances from these bacteria or foods has been attempted. Many attempts have been made to control cancer with bacteria, such as Coley's toxin (1964) using culture filtrates of Serratia and streptococci, and treatment of leukemia with BCG (Mathe, G. et al. Cancer Res., 14, 1, 1971) and regression of cancer masses in guinea pigs (Zbar, B., et al., J. Natl. Cancerlnst., 48, 831, 1971), and yeast wall polysaccharide. Has been reported to be effective against transplanted cancers such as sarcoma 180 by administration of sarcoma.
  • Shiitake mushrooms are edible mushrooms that represent Japan and China and have been artificially cultivated in Japan for about 300 years, but their pharmacological effects and medicinal components have recently been elucidated.
  • tumor cells such as large intestine and liver in rats and mice
  • mitogen fruit Tabata, T. et al., Immunopharmacology, 24: 57, 1992; Hibino, et al., Immunopharmacology, 28: 77, 1994.
  • the present inventors focused on the LAK activity enhancing effect (anti-tumor and Z or anti-cancer activity) of Shiitake, and examined the in vivo LAK activity enhancing effect when Shiitake mycelium extract was directly administered to a living body. It is an object to provide a screening substance and a method thereof for screening in vitro and in vitro.
  • the LAK activity enhancer is actually administered in vivo, Alternatively, prepare a large amount of in vivo lymphocytes, activate them in vitro with a LAK activity enhancer, and then return the activated lymphocytes to the living body to determine if they have the LAK activity enhancement effect. Had been confirmed. In addition to the drawback that this method is too expensive for treatment, there is a drawback that the physical burden on the subject becomes a problem. Therefore, if the LAK activity enhancer can be simply screened in vitro as to whether or not it actually has an effect in a living body, the physical and financial burden on the subject can be greatly reduced. Disclosure of the invention
  • the present inventors have found that components extracted from hyphae, which is a form in front of the edible body in the life cycle of shiitake mushrooms, have an immunostimulatory activity far superior to that of the body, and have antitumor activity and Z or anticancer activity. I found it to be active.
  • the extract as a substitute for IL-2 as a LAK activity inducer in vitro, the antitumor effect and / or anti-tumor effect in vivo when the extract is directly administered to a living body.
  • the present inventors have found that a cancer action, particularly an LAK activity enhancing action, can be squelched in vitro, and have completed the present invention.
  • the present inventors have developed an in vivo cytotoxic activity when an antitumor substance or an anticancer substance containing a shiitake mycelium extract, particularly a LAK activity enhancer is directly administered to a living body.
  • a LAK activity enhancer has a positive correlation with the cytotoxic activity when lymphocytes prepared from a subject are activated in vitro with the aforementioned LAK activity enhancer.
  • the present invention comprises the following steps:
  • a method for in vitro determining a substance having a LAK activity enhancing action suitable for a subject comprising:
  • the present invention also relates to the in vitro screening method described above.
  • the present invention provides a screening substance containing a shiitake mycelium extract, which can screen whether LAK activity can be enhanced in vivo. Therefore, the present invention provides a mycelium extract of shiitake mushroom, which can be determined in vitro before administration of a LAK activity enhancer, whether or not the LAK activity enhancer can be expected to have an in vivo LAK activity enhancer effect.
  • the present invention relates to a contained screening substance and a screening method thereof.
  • the screening substance and the screening method of the present invention can be used not only for humans but also for livestock.
  • LAK activity refers to the antitumor activity of cytotoxic T lymphocytes that can attack tumors that cannot be recognized by lymphocytes that carry NK activity and have little effect on normal cells of the self. Means that. “Enhancement of LAK activity” refers to enhancing such LAK activity, which means inducing LAK cells from lymphocytes or further increasing the antitumor activity of already existing LAK cells.
  • the antitumor activity of LAK cells can be increased. This leads to an enhancement of the cellular immune system. Therefore, it can be used for treatments aimed at improving the immune system, in addition to treatments expected to improve antitumor activity.
  • FIG. 1 is a bar graph showing the results of Table 1 showing the results of LAK activity enhancement screening using the shiitake mycelium extract of the present invention.
  • the present invention relates to whether an antitumor or anticancer agent containing a Shiitake mycelium extract, particularly a preparation for enhancing LAK activity, can be expected to enhance LAK activity by directly administering it to a living body.
  • a screening substance containing a mycelium extract of Shiitake mushroom which can be determined in vitro before administration of the preparation, and a method for screening the same.
  • the “screening substance” refers to a substance used for in vitro testing the effect of enhancing LAK activity in vivo when administered to a living body.
  • “Shitake mushroom cell extract” used as a screening substance is obtained by cultivating Shiitake fungi on a solid medium. It refers to an extract obtained by crushing and decomposing mycelium or a solid medium containing mycelium obtained in the presence of water and enzymes.
  • the shiitake mycelium extract is preferably obtained by the following method.
  • the force is not limited to this. That is, after inoculating shiitake mushroom on a solid medium based on bagasse (sugar cane squeezer) and defatted rice bran and growing the mycelium, the solid medium in which the mycelium had grown was passed through 12 mesh. Unbundle to 30% by weight or less. Water is added to the dissociated solid medium, and then carbohydrate-degrading enzyme, proteolytic enzyme, or enzyme (s) comprising a combination thereof are added, and the temperature is maintained at 30 to 35 ° C. The solid medium is ground and crushed.
  • Enzymes used in this step include, but are not limited to, cellulases, proteases, and dalcosidases.
  • the solid medium crushed and crushed in the above step is prepared so that at least 70% by weight or more of the bagasse fiber can pass through the 12 mesh, and then the enzyme is heated to a temperature of up to 95 ° C to reduce the enzyme. Inactivate and sterilize at the same time. Finally, the obtained suspension liquid is filtered to obtain a shiitake mycelium extract.
  • the shiitake mycelium extract may be used as it is as the screening substance or the immunotherapeutic agent of the present invention, but it is convenient to concentrate, freeze-dry and store it as a powder and use it in various forms when used. It is.
  • the powder obtained by freeze-drying is a brown powder, hygroscopic and has a unique taste and odor.
  • the shiitake mycelium extract of the present invention can be added directly to a lymphocyte fraction obtained from peripheral blood.
  • the concentration of the shiitake mycelium extract contained in the screening substance of the present invention is preferably 1 ng / ml to 100 mg / ml, more preferably 1 ng / ml to 100 mg / ml. zg / ml to 100; g / ml, particularly preferably 10 ig / ml to 50 g / ml.
  • the shiitake mycelium extract of the present invention is preferably subjected to sterile treatment with acetone before being added to cultured cells or directly added to peripheral blood.
  • the method of screening using the screening substance of the present invention is according to the method of Takagi et al. (Clinical Immunity, 19: 245-249, 1987), but instead of IL-2, a screening substance, for example, This method differs from the conventional method in using the shiitake mycelium extract of the present invention. That is, the screening method for enhancing LAK activity of the present invention comprises the following steps:
  • the LAK induction sample is prepared, for example, by the following method. That is, the lymphocyte fraction obtained by way described above, the final concentration of cells is prepared so as to IX 10 5 / ml ⁇ ixi0 6 / ml , suspended in the culture solution, the cells per Ueru Adjust the cell count to between 1 ⁇ 10 4 / ⁇ to 1 ⁇ 10 5 / ⁇ by adding 100 1 of the suspended culture.
  • the number of cells per well can be appropriately determined by those skilled in the art according to the activity of the effector cells to be used, the sensitivity of the target cells to effecuta cells, and the like.
  • the suspension is supplemented with a shiitake mycelium extract at a final concentration of 1 ng / ml to 100 mg / ml as a screening substance according to the experimental design.
  • the subject's lymphocytes are cultured in the presence of various concentrations (including zero concentration) of the shiitake mycelium extract of the present invention to obtain effector cells.
  • effector cells refers to cells that have been subjected to the above-described culture treatment for 3 days, and are lymphocytes cultured with a LAK activity enhancer (reagents treated with an LAK activity enhancer). Lymphocytes cultured in culture medium without LAK activity-enhancing substances only (lymphocytes treated without induction).
  • Control samples are prepared in the same manner as for the preparation of LAK-derived samples, except that sterile recombinant IL-2 (rIL-2; 2000 l / ml) is added instead of adding the screening substance.
  • rIL-2 sterile recombinant IL-2
  • the measurement of LAK activity is carried out by a method such as 51 Cr-release Atssay method and [] lysine method.
  • a method such as 51 Cr-release Atssay method and [] lysine method.
  • the 51 Cr-releasing assay is a method for measuring in vitro the target cytotoxic activity of LAK cells derived from a lymphocyte treated with a LAK activity enhancer.
  • the 5'Cr release technology involves the following steps:
  • effector cells eg, killer T cells or LAK cells
  • a Daudi cell or a Raji cell is preferably used as a subcultured cell which is a target cell used in the 5 l Cr-releasing assay.
  • Culture the target cells in a culture flask collect them, label them with 51 Cr, and dispense them into a microtiter plate.
  • the culture solution of the target cells use a culture solution suitable for the growth of the cells to be used. For example, use a culture solution such as RPMI 1640 to which serum, antibiotics, etc. are appropriately added.
  • Labeled target cells were added to 51 Cr- chromic acid Natoriumu of 100 to 150 ⁇ Ci to target cells 10 per six cultured, carried out by 1-2 hours Inkyubeto be Rukoto at 37 ° C for after stirred well .
  • Cultured cells were washed 3 times with PBS, and used those suspended in 10% FBS added RPMI 1640 medium so as to ixi0 6 / ml.
  • FBS fetal calf serum
  • FCS fetal calf serum
  • the target cytotoxic activity in 51 Cr-releasing assays is expressed by the following formula: Experimental dissociation (cpm) —Spontaneous dissociation (cpm)
  • the radiation dose due to 5lCr released into the culture supernatant can be measured by using a scintillation counter or the like.
  • each step is performed as follows, but those skilled in the art will recognize that appropriate changes and modifications may be made. Collect the culture supernatant of each well from the cultured plate, and measure the radioactivity by using a set-up counter.
  • the Shiitake mushroom mycelium extract which was confirmed to have LAK-inducing activity in inViIro by the above screening method, was administered to a living body at 3600 mg / day for 7 days to induce LAK activity in invitro.
  • the lymphocyte fraction collected by the lymphocyte collection method described above the LAK activity% was measured under the same conditions as for the LAK-induced sample described above. It was found to show a positive correlation with the results obtained in.
  • a crushing and crushing action is applied to a portion of the solid culture medium for about 200 minutes so that about 80% by weight of the bagasse fiber passes through the 12 mesh. I made it.
  • the pulverization and crushing of the medium-containing mixture were performed while gradually increasing the temperature of the mixture.
  • the mixture containing the medium was further heated to 90 ° C. to inactivate the enzyme, and at the same time, sterilized, and left at 90 ° C. for 30 minutes.
  • the obtained medium-containing mixture was filtered through a 60-mesh filter cloth to obtain a shiitake mushroom mycelium extract, which was concentrated to obtain a lyophilized powder.
  • the shiitake mushroom mycelium extract obtained in this way has a phenol-sulfuric acid method 25.3% of carbohydrate (w / w) by quality analysis, 19.7% (w / w) of protein by Raleigh protein analysis, and 2.6% (w / w) of polyphenol by Folon-Denis method based on gallic acid Weight) included.
  • the Shiitake mycelium extract also contained 8% crude fat, 22% crude ash, and about 20% soluble nitrogen-free substances other than carbohydrates.
  • the constituent sugar composition in the shiitake mycelium extract was as follows. Xyl: 15.2; Ara: 8.2; Man: 8.4; Gul: 39.4; Gal: ⁇ .4; GlcN ": 12.0; GLuUA: 11.3
  • Example 2 Measurement of LAK activity
  • peripheral blood before taking Shiitake mycelium extract from subjects A, B, and C, and peripheral blood after oral administration of 1200 mg of Shiitake mycelium extract to each subject three times daily for one week Blood was collected respectively.
  • lymphocyte fraction separated from the peripheral blood by the following method, the ability of the extract of the present invention to activate lymphocytes in vivo and the use of the extract to It can be screened for correlation with its ability to activate in vitro.
  • plasma autologous serum
  • FBS RPMI 1640 subculture cells are the target cells cultured in culture in (Daudi cells) were harvested by centrifugation, 10 6 cells per 100 ⁇ 150 ⁇ ( ⁇ of 51 Cr- sodium chromate (New England Nuclear) was added, incubated for one hour at 37 ° C with 5% C0 2 incubator. 5 'after three times washing the labeled cell cultures with PBS by Cr, 10% so as to 1X10 6 / ml The cells were suspended in RPMI 1640 culture medium supplemented with FBS.
  • 100 l each the spontaneous dissociation group (negative control) was dispensed with 100 l of RPMI 1640 culture medium supplemented with 10% FBS, and the experimental dissociation group was dispensed with 10 xg / ml of the present invention.
  • Effector cells 100/1 (1 ⁇ 10 4 / well) each) stimulated with bamboo mycelium extract or rIL-2 at a concentration of 2000 ⁇ l / ml as a control were dispensed. After collecting the cells Ueru bottom and centrifuged for 5 minutes at 800 rpm by a plate centrifugal separator, and cultured for 3.5 hours at 37 ° C with 5% C0 2 incubator.
  • the culture supernatant of each well was collected from the cultured plate using S0KEN-PET No.-96, and the radioactivity was measured by a scintillation counter.
  • LAK activity was calculated according to the following table. '' Experimental dissociation (cpm) Natural dissociation (cpm)
  • Subject A Subject B Subject C Before taking 13% 27% 14% Screening with extract
  • the extract of the present invention can be obtained by stimulating lymphocytes collected from the peripheral blood of the subject C using the extract of the present invention.
  • the effect of enhancing LAK activity obtained by direct oral administration was not observed (see Table 1, Experiment 2). Therefore, it was also found from this example that the effect of enhancing the LAK activity in vivo when the Shiitake mushroom mycelium extract of the present invention was orally administered can be predicted by screening with in Vitr0.
  • the LAK activity enhancing effect in vitro when the LAK activity enhancing substance of the present invention is orally administered to a living body can be more accurately predicted from the screening results in vitro.
  • the LAK activity enhancing effect in vivo when the Shiitake mycelium extract of the present invention is directly administered can be easily determined in vitro before directly administering the LAK activity enhancing substance,
  • LAK activity can be enhanced to a subject that cannot be expected to enhance LAK activity.
  • Useless administration of the substance can be prevented.
  • the method of the present invention allows the in vivo therapeutic effect of the LAK activity enhancer to be screened in vitro without collecting a large amount of lymphocytes from the blood of the subject.
  • the burden is also significantly reduced.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • General Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Urology & Nephrology (AREA)
  • Mycology (AREA)
  • Biotechnology (AREA)
  • Hematology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Cell Biology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Food Science & Technology (AREA)
  • Toxicology (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Botany (AREA)
  • Medical Informatics (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

L'invention concerne une substance bon marché destinée à cribler un agent immuno-thérapeutique ayant une activité anti-tumorale et/ou anti-cancéreuse. Elle concerne une méthode qui permet de déterminer in vitro une substance ayant un effet approprié de potentialisation de l'activité LAK chez un sujet. La méthode consiste à: a) recueillir le sang périphérique du sujet et préparer une fraction lymphocytaire de ce sang; b) préparer, d'une part, un prélèvement d'induction de LAK par adjonction de la substance de criblage à la fraction lymphocytaire, d'autre part, un prélèvement de contrôle exempt de substance de criblage; et c) mesurer l'activité LAK dans le prélèvement d'induction et l'activité LAK dans le prélèvement de contrôle, et comparer ces activités afin de déterminer l'effet de potentialisation de l'activité LAK de la substance de criblage in vitro sur ledit sujet.
PCT/JP1999/006615 1998-11-27 1999-11-26 Substance contenant un extrait d'hypes du champignon shiitake destinee a cribler l'activite lak, et procede mettant en oeuvre ladite substance pour cribler l'activite lak WO2000033069A1 (fr)

Priority Applications (5)

Application Number Priority Date Filing Date Title
CA002352614A CA2352614C (fr) 1998-11-27 1999-11-26 Substance contenant un extrait d'hypes du champignon shiitake destinee a cribler l'activite lak, et procede mettant en oeuvre ladite substance pour cribler l'activite lak
GB0112999A GB2360090B (en) 1998-11-27 1999-11-26 Lak activity-screening materials containing Lentinus extract of edodes mycelium and lak activity-screening methods using the extract
KR1020017006475A KR20010089498A (ko) 1998-11-27 1999-11-26 표고버섯 균사체 추출물을 함유하는 ｌａｋ 활성스크리닝물질 및 이를 사용한 ｌａｋ 활성스크리닝법
HK02101648.2A HK1041518B (zh) 1998-11-27 2002-03-04 含有香菇菌絲體提取物的淋巴因子活化殺傷細胞活性-過篩材料以及使用該提取物的淋巴因子活化殺傷細胞活性-過篩方法
HK02104333.6A HK1042745A1 (zh) 1998-11-27 2002-06-08 含有香菇菌絲體提取物的lak活性篩選材料以使用該提取物的lak活性篩選方法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP35392698A JP4308350B2 (ja) 1998-11-27 1998-11-27 シイタケ菌糸体抽出物を含有するｌａｋ活性スクリーニング物質およびそれを用いたｌａｋ活性スクリーニング法
JP10/353926 1998-11-27

Publications (1)

Publication Number Publication Date
WO2000033069A1 true WO2000033069A1 (fr) 2000-06-08

Family

ID=18434161

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP1999/006615 WO2000033069A1 (fr) 1998-11-27 1999-11-26 Substance contenant un extrait d'hypes du champignon shiitake destinee a cribler l'activite lak, et procede mettant en oeuvre ladite substance pour cribler l'activite lak

Country Status (8)

Country Link
JP (1) JP4308350B2 (fr)
KR (1) KR20010089498A (fr)
CN (1) CN1314963C (fr)
CA (1) CA2352614C (fr)
GB (1) GB2360090B (fr)
HK (2) HK1041518B (fr)
TW (1) TWI222515B (fr)
WO (1) WO2000033069A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014008353A2 (fr) 2012-07-05 2014-01-09 Nutramax Laboratories, Inc. Compositions comprenant du sulforaphane ou un précurseur de sulforaphane et un extrait ou une poudre de champignon
WO2022251524A1 (fr) 2021-05-26 2022-12-01 Nutramax Laboratories, Inc. Compositions comprenant du sulforaphane ou un précurseur de sulforaphane et des constituants de plant de moringa

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103173379A (zh) * 2011-12-22 2013-06-26 熊津食品有限公司 新型的红参发酵用微生物，及其利用该微生物的红参发酵液和红参发酵饮料
JP5847106B2 (ja) 2013-03-25 2016-01-20 株式会社フジキン 流量モニタ付圧力式流量制御装置。

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2015294A1 (fr) * 1989-04-28 1990-10-28 John J. Rinehart Generation et expansion de cellules lak
US5108760A (en) * 1989-07-21 1992-04-28 Terumo Corporation Enhances lak cell activation by treatment of human peripheral blood mononuclear cells with amino acid amides
US5198334A (en) * 1989-07-21 1993-03-30 Terumo Corporation Protection of natural killer cell cytolytic activity in peripheral blood mononuclear cells
US5725855A (en) * 1991-04-05 1998-03-10 The United States Of America As Represented By The Department Of Health And Human Services Method of treating tumors with CD8+ -depleted or CD4+ T cell subpopulations
JP2002270532A (ja) * 2001-03-14 2002-09-20 Tokyo Electron Ltd 加熱装置及び熱処理装置

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
K. AMINO, "Hito rinpakyu ni taisuru renchinan no menneki fukatsu sayo ni tsuite", Gan to Kagaku Ryoho, (1981), Vol.8, No.6, pages 967-973. *
LAGADEC P.F., "Human smell-cell lung-cancer cells are cytokine-resistant but nk-lak-sensitive", International journal of cancer (1991), vol. 48, no. 2, pages 311-317. *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014008353A2 (fr) 2012-07-05 2014-01-09 Nutramax Laboratories, Inc. Compositions comprenant du sulforaphane ou un précurseur de sulforaphane et un extrait ou une poudre de champignon
EP3666277A1 (fr) 2012-07-05 2020-06-17 Nutramax Laboratories, Inc. Compositions comprenant du sulforaphane ou un précurseur de sulforaphane et un extrait ou une poudre de champignon
WO2022251524A1 (fr) 2021-05-26 2022-12-01 Nutramax Laboratories, Inc. Compositions comprenant du sulforaphane ou un précurseur de sulforaphane et des constituants de plant de moringa

Also Published As

Publication number Publication date
CN1332847A (zh) 2002-01-23
GB0112999D0 (en) 2001-07-18
CN1314963C (zh) 2007-05-09
GB2360090A (en) 2001-09-12
JP4308350B2 (ja) 2009-08-05
JP2000162204A (ja) 2000-06-16
KR20010089498A (ko) 2001-10-06
TWI222515B (en) 2004-10-21
GB2360090B (en) 2004-03-24
HK1041518A1 (en) 2002-07-12
CA2352614C (fr) 2009-03-10
HK1041518B (zh) 2005-03-24
CA2352614A1 (fr) 2000-06-08
HK1042745A1 (zh) 2002-08-23

Similar Documents

Publication Publication Date Title
US20190276803A1 (en) Method of culturing immune cells, kit for thereof, immune cell cultured medium obtained by same method, cosmetic composition and pharmaceutical composition comprising thereof
KR101467903B1 (ko) 흑미의 담자균류균사 발효 및 생물전환공정을 통해 생산된 면역증강제
Kim et al. Dendritic cell activation by polysaccharide isolated from Angelica dahurica
Zhu et al. Anti-tumor and immunomodulatory activities of an exopolysaccharide from Rhizopus nigricans on CT26 tumor-bearing mice
JP2016539184A (ja) 免疫調節剤としての用いるための藻類抽出物
TWI235764B (en) Method of extensive culture of antigen-specific cytotoxic T cells
JP2002204687A (ja) β−１．３−１．６グルカン（アウレオバシジウム培養液）の医療、保健、福祉、食品および各種産業分野での応用
CN102319426B (zh) 一种去除调节性t细胞的特异性肿瘤疫苗的制备方法
Kodama et al. Potential antitumor activity of a low-molecular-weight protein fraction from Grifola frondosa through enhancement of cytokine production
JP2024050573A (ja) 腫瘍浸潤リンパ球（ｔｉｌｓ）を活性化する方法
WO2000033069A1 (fr) Substance contenant un extrait d'hypes du champignon shiitake destinee a cribler l'activite lak, et procede mettant en oeuvre ladite substance pour cribler l'activite lak
CN114507640B (zh) 一种高增殖能力和高细胞毒性cik细胞的培养方法及其应用
JP2006124383A (ja) 樹状細胞活性化剤
JP7088502B2 (ja) 抗体医薬の効果増強用組成物
US6919081B1 (en) Lak activity potentiator orginating in shitake mushroom hyphae extract and lak activity potentiating preparations containing the same
JP2000159686A (ja) シイタケ菌糸体抽出物由来のｌａｋ活性増強用製剤
Takahashi et al. Biological activity of Pityrosporum. II. Antitumor and immune stimulating effect of Pityrosporum in mice
WO2019209706A2 (fr) Composés fongiques bioactifs produits par fermentation à l'état solide
JP2004099613A (ja) 自己免疫疾患治療用ガノデルマルシダム胞子
JP2000159685A (ja) シイタケ菌糸体抽出物由来のｌａｋ活性増強物質
CN106282109A (zh) 一种针对肝癌的dc‑ctl试剂盒及其制备方法
JP2006141346A (ja) 樹状細胞活性化剤
Abutu et al. Regulatory activity of ethanol leaves extract of Moringa Oleifera on benzene induced leukemia in Wister Rat using TNF-α analysis
JP2022187488A (ja) ヒメマツタケ菌糸体亜臨界水抽出物の製造方法
KR20240011367A (ko) 면역증진 효능이 개선된 땅콩새싹 추출물

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 99815189.0

Country of ref document: CN

AK Designated states

Kind code of ref document: A1

Designated state(s): CA CN GB KR SG US

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: 1020017006475

Country of ref document: KR

ENP Entry into the national phase

Ref document number: 2352614

Country of ref document: CA

Ref document number: 2352614

Country of ref document: CA

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 200112999

Country of ref document: GB

Kind code of ref document: A

WWP Wipo information: published in national office

Ref document number: 1020017006475

Country of ref document: KR

WWE Wipo information: entry into national phase

Ref document number: 09856716

Country of ref document: US

WWR Wipo information: refused in national office

Ref document number: 1020017006475

Country of ref document: KR