WO2005054496A1 - Méthode d'examen de la cinétique des cellules - Google Patents

Méthode d'examen de la cinétique des cellules Download PDF

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WO2005054496A1
WO2005054496A1 PCT/JP2004/017843 JP2004017843W WO2005054496A1 WO 2005054496 A1 WO2005054496 A1 WO 2005054496A1 JP 2004017843 W JP2004017843 W JP 2004017843W WO 2005054496 A1 WO2005054496 A1 WO 2005054496A1
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cancer
cells
immunotherapeutic agent
perforin
cancer immunotherapeutic
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PCT/JP2004/017843
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English (en)
Japanese (ja)
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Akikuni Yagita
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Orient Cancer Therapy Co., Ltd.
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Publication of WO2005054496A1 publication Critical patent/WO2005054496A1/fr

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    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention provides a new area of cancer treatment.
  • a tyrosine kinase inhibitor which is attracting attention as a novel cancer treatment, and NK cell activation, NKT cell activation, angiogenesis inhibitory, and IL-12 production induction developed by Dr. New immunotherapy that focuses on the dynamics of perforin-producing cells in combination with new immunotherapy that focuses on the dynamics of IFN ⁇ and the ability to induce the production of IFN ⁇ . .
  • IL-12 interleukin 12
  • MTC Novel Immunotherapy for cancer
  • IL-12 has TNF a ⁇ IFN y ⁇ IL-12 ⁇ CTL activity, and has an activating and enhancing effect on killer T cells through the root.
  • the enhancement of IL-12 production is expected to have an anticancer effect by activating and enhancing killer T cells.
  • Non-patent Document 1 NKR-P1; natural killer receptor P1
  • NKR-P1 ⁇ cell antigen receptor
  • Non-patent Document 2 Yagida has found that NKR-P1 is also involved in the activity of ⁇ cells, and that this activity is more superior in anticancer effect (Patent Document 2).
  • the molecular target therapeutic agent for cancer has attracted attention as a new type of anticancer agent in comparison with the conventional cell target therapeutic agent.
  • a tyrosine kinase inhibitor has been particularly noted as a drug having a signal transduction inhibitory action.
  • ZD1839 (Iressa: registered trademark AstraZene Power) has a competitive action with EG at the binding site of EGFR (Epidermal Growth Factor Receptor) tyrosine kinase, and suppresses tyrosine kinase autophosphorylation by controlling tyrosine kinase. Suppress activity.
  • signal transduction related to the proliferation, invasion, differentiation, and metastasis of EGFR (the binding of EGFR tyrosine kinase to the extracellular domain of EGFR Activating, triggering autophosphorylation of EGFR and phosphorylation of various intracellular target proteins transmit cell surface forces, proliferation signals to nuclei, and proliferate signals from cancer cell surfaces to nuclei. Is transmitted to cause cancer cell proliferation, invasion, metastasis, and angiogenesis.
  • IMC-C225 EGFR-targeted monoclonal antibody
  • Herceptin is a monoclonal antibody against Her2 / Neu with homology to EGFR, and STI-571 (Gleevec) is a tyrosine kinase activity of BCR-Abl And the ability to inhibit the tyrosine kinase activity of c kit (Non-Patent Document 2).
  • ZD1839 is a powerful and selective EGFR tyrosine kinase inhibitor newly developed by AstraZeneca, and its usefulness has been proven in humans.
  • Clinical results for non-small cell lung cancer and prostate cancer have PR (partial remission) of 10 to 20% and CR (complete remission) at all, but they are very rare It took more than four months to complete remission. Therefore, combination therapy with ZA1839 (Iressa) and various anticancer drugs has been tried! /, But at the moment it is additive !, but no synergistic effect has been obtained.
  • CX3C-chemokine has been reported as a chemokine that induces IFN ⁇ production from NK cells, and its receptor is CX3CR1, which is a G protein-coupled seven-transmembrane receptor ⁇ cells, monocytes and some of them). It is expressed in cells (Non-Patent Document 3). Nishimura et al. Have shown that CX3CR1 expression in ⁇ cells is limited to CD3 (+) CD161 (+) and granzym ⁇ positive cells, and that cytotoxic activity is carried by CX3CR1 positive cells (Non-patent Document 4 )
  • Patent Document 1 JP-A-10-139670
  • Patent Document 2 US2002-0010149A1
  • Non-patent document 1 Special issue: Basics and clinical practice of NKT cells: Latest Medicine 55 Vol. 4 2000 818—823 ⁇ 1 ⁇
  • Non-Patent Document 2 Blood and Immune 'Tumor Vol. 7 No.3 2002-7
  • Non-Patent Document 3 Cell 1997; 91: 521
  • Non-Patent Document 4 J. Immunol 2002; 168: 6173
  • the present invention aims to provide a more effective effect of the above-mentioned molecular target therapeutic agent, and achieves a higher complete remission rate, a shorter time to complete remission, and a synergy with immunotherapy. It provides the means to achieve the effect.
  • new immunotherapy focusing on CTL activity, NKT activity, NK activity, VEGF, etc., and molecular targeted therapeutics, especially tyrosine kinases
  • the task is to achieve a synergistic effect when used in combination with an inhibitor.
  • the cancer immunotherapy in order for a combined use of a tyrosine kinase inhibitor and a cancer immunotherapy to achieve an excellent synergistic effect in cancer treatment, the cancer immunotherapy effectively affects perforin-producing cells. It has been found that this is important, and the present invention has been completed.
  • a cancer immunotherapeutic agent that uses the dynamics of perforin-producing cells as a marker to determine the efficacy of the cancer immunotherapeutic agent in combination with a tyrosine kinase inhibitor and a cancer immunotherapeutic agent.
  • the tyrosine kinase inhibitor has a selective targeting effect on at least one of the following receptors, and the function of the cancer immunotherapeutic agent is IL-12 production induction and Z or IL-21-mediated response. 3 ways.
  • HER2 / neu HER3, HER4, c-kit, PDGFR, bcr-abl, EGFR
  • the tyrosine kinase inhibitor has a selective targeting effect on at least one of the following receptors, and the function of the cancer immunotherapeutic agent is IL-12 production induction and Z or IL-21-mediated response. Cancer immunotherapy.
  • HER2 / neu HER3, HER4, c-kit, PDGFR, bcr-abl, EGFR
  • the cancer immunotherapeutic agent according to the above item 5 wherein the cancer immunotherapeutic agent is a substance having a
  • the cancer immunotherapy agent is a yeast-derived component having a ⁇ 1,3 / 1,6 glucan structure.
  • the cancer immunotherapy agent according to the above item 5 wherein the cancer immunotherapy agent is a yeast-derived component having a ⁇ 1,3 / 1,6 glucan structure.
  • 10.Perforin-producing cell dynamics are determined by at least one of NKTP (+), NKT8 (+) P (+), CD8 (+) P (+) T, and Thl / Th2 ratio Any one of methods 1, 3, 4, 6, and 8 above.
  • NKTP (+), NKT8 (+) P (+), CD8 (+) P (+) T, and Thl / Th2 ratio The cancer immunotherapeutic agent according to any one of the above items 2, 5, 7, and 9.
  • FIG. 1 is a multivariate analysis of the contribution of an immune marker.
  • FIG. 2 is a diagram in which the contribution of perforin-producing cells to each effector cell was examined.
  • FIG. 3 is a diagram showing the degree of contribution as a ratio.
  • FIG. 4 is a diagram examining the importance of immunological factors.
  • FIG. 5 is a diagram examining the importance of immunological factors.
  • FIG. 6 is a diagram examining the importance of immunological factors.
  • FIG. 7 is a diagram examined for NK cells.
  • FIG. 8 is a diagram examined with CD8 (+) cells.
  • Dr. Yagida's New Cancer Therapy for Cancer is a therapeutic means comprising a combination of four different mechanisms of action.
  • the first mechanism of action is to administer an angiogenesis inhibitor (Bettershark) to impede blood flow to the cancer and to reduce the size of the cancer.
  • the effect can be determined by measuring vascular endothelial cell growth factor (VEGF).
  • VEGF vascular endothelial cell growth factor
  • the angiogenesis inhibitory effect can be evaluated by a negative (negative) VEGF value (-VEGF). It is possible to evaluate the angiogenesis inhibitory ability by using other vascular growth factors such as FGF and HGF instead of the VEGF value.
  • the positive value of an angiogenesis inhibitor can be evaluated instead of VEGF (for example, endstatin value).
  • the first mode of action is that a compound having a ⁇ 1,3 glucan structure is administered to induce Thl cytokines (TNF o; IFN y, IL-12) to activate CTLs.
  • TNF o Thl cytokines
  • This CD8 (+) perforin level includes cytotoxic T cells (CTL) and immunosuppressive T cells (STC; Suppressor T cells).
  • CTL cytotoxic T cells
  • STC immunosuppressive T cells
  • IFN y force S 10 IU / ml or more or the IL-12 value is 7.8 pg / ml or more, it is CTL, and if the IFN ⁇ and IL-12 are low, it is judged as STC. Therefore, CTL activity can be evaluated based on IFN y production ability (IFN y value) or IL-12 production ability (IL-12 value).
  • the effector cells activated by the administration of the compound having the ex-1,3 glucan structure, which is the third and fourth mechanism of action, are NK cells and NKT cells.
  • the NK and NKT cells share the NKR-P1 (NK cell receptor CD161 (+), and for the former, the number of NK cells can be measured using the CD3 (-) CD161 (+) surface marker.
  • Activatedi can be determined by its ability to produce CD3 (-) CD161 (+) perforin, whereas the latter NKT cells are CD3 (+) CD161 (+).
  • the cell number can be measured, and the activity of NKT cells can be measured by its perforin-producing ability (denoted as NKTP (+)).
  • NITC new immunotherapy
  • general immunotherapy it is possible to evaluate each effector cell or angiogenesis inhibitory effect by the following measurement items. It is possible. Specifically, CTL activity can be evaluated based on its ability to induce IFNy or IL-12 production.
  • NK cell activity is CD3 (-) CD161 (+), or
  • Evaluation can be performed using CD3 (+) CD161 (+) or CD3 (+) CD161 (+) perforin (NKTP).
  • the present invention was carried out by examining clinical results obtained by using a tyrosine kinase inhibitor in combination with the above-mentioned new immunotherapy.
  • the present inventors have proposed a new immunotherapy (NITC) in cancer patients, a compound carrying an a1,3 glucan structure, a compound carrying a / 1,3 glucan structure, and an angiogenesis inhibitory substance (shark cartilage).
  • NITC new immunotherapy
  • a compound carrying an a1,3 glucan structure a compound carrying a / 1,3 glucan structure
  • an angiogenesis inhibitory substance harvested cartilage
  • IL-12, IFNy, and various other sites were measured.
  • CD8 (+) perforin production has a strong positive correlation with IFNy and IL-12 production.As a result, measurement of CD8 (+) perforin production is significant for the evaluation of CTL activity route. Have found that there is.
  • a tyrosine kinase inhibitor is used in combination with a new
  • At least the measurement of perforin-producing cells can be applied to a method for screening a cancer immunotherapy agent useful in combination with a tyrosine kinase inhibitor. Screening methods can be used to demonstrate useful cancer cytotoxicity in combination with tyrosine kinase inhibitors. It is possible to identify the new ⁇ 8 1,3 glucan to be carried.
  • the cancer immunotherapeutic agent used in the present invention is, for example, a mushroom mycelium composition preparation having a ⁇ 1,3 glucan structure (eg, ILX ffi ⁇ : Tozai Pharmaceutical Research Institute, ILY ffi ⁇ : Seishin Enterprise), or j8 Various yeasts with 1,3 glucan structure (marine yeast, baker's yeast, NBG TM) can be used. In particular, marine yeast is preferred.
  • a novel cancer immunotherapeutic agent can be easily identified by those skilled in the art by combining measurement of perforin-producing cells. According to this method, in the present invention, marine yeast was most suitable as a useful cancer immunotherapeutic agent in combination with a tyrosine kinase inhibitor, and it was considered that this system also activated CX3CR1-expressing cells.
  • this cancer immunotherapy agent a combination use of this cancer immunotherapy agent and a tyrosine kinase inhibitor is essential.
  • various oral synthase inhibitors using ZD1839 (trade name of Iressa) or STI571 (trade name of Dalibec) can be effectively used.
  • target molecules include HER2 / neu, HER3, HER4, c-kit, PDGFR, bcr-abl, EGFR and the like.
  • the most effective molecules are EGFR or C-kit.
  • the dose of the tyrosine kinase inhibitor follows the recommended dose of each molecular target compound,
  • Oral administration of 10-500 mg / day is performed.
  • the combination use of the cancer immunotherapeutic agent and the tyrosine kinase inhibitor is not particularly limited, but may be performed from the early stage of the treatment or either of them may be performed first.
  • a dramatic clinical effect was confirmed using a tyrosine kinase inhibitor in combination with NITC therapy, especially a cancer immunotherapy drug, for a certain period of time.
  • an NK activator or NKT activator can be used in combination with an IL-12 production inducer as a cancer immunotherapy agent.
  • - Geroorigo sugar is useful as a 1, 3-glucan composition formulations structure with compound NK or NKT activator agent such Fukoidan.
  • a Various compounds having a 1,3 glucan structure are known, and the known structure and CD3 (-) CD161 (+), CD3 (-) CD161 (+) perforin-producing ability, CD3 (+) CD161 (+), Those skilled in the art can easily specify the NK-activating agent by combining the measurement of the ability to produce CD3 (+) CD161 (+) perforin.
  • CD3 (+) CD161 (+) means that it acts on the NKT cell receptor NKR-P1.
  • saccharide substances having a 1,3 glucan structure examples include -gelooligosaccharide (TSO), fucoida , Sulfated oligosaccharides and the like.
  • -Gello-oligosaccharide is a saccharide containing 3-0-a D darcoviranosyl-D glucose as a constituent unit.
  • Typical examples include -gelose, -gelosylglucose, and digerosylmaltose.
  • Examples of commercially available -gelooligosaccharides include -gerooligosaccharide liquid sugar (seller: Takeda Shokuhin Kogyo Co., Ltd.).
  • Fucoidan in a narrow sense, is a sulfated fucose-containing polysaccharide in which one to two sulfuric acid molecules are bonded to two to six molecules of fucose, and a fucoidan-like polysaccharide containing xylose or peronic acid is called ⁇ fucoidan '' at the food level. Is called. Fucoidan, for example, is obtained by crushing kelp, chipping, extracting aqueous components, removing the extraction residue by centrifugation, removing low-molecular substances such as eodo and sodium salt by ultrafiltration, and freeze-drying. It is formulated into a formulation.
  • fucoidans examples include fucoidan derived from brown algae, such as fucoidan derived from gagome kelp, and fucoidan derived from Okinawa mozuku.
  • Fucoidans derived from brown algae Laminariaceae, such as gagome kelp include at least three types of fucoidan, F-fucoidan (a polymer of a L-fucos), and U-fucoidan (13D-glucuronic acid and a-D mannose as the main chain and a L-fucose) and G-fucoidan (
  • F-fucoidan a polymer of a L-fucos
  • U-fucoidan 13D-glucuronic acid and a-D mannose as the main chain and a L-fucose
  • oligosulfate sulfate examples include, for example, an extract derived from susabinori (Poryphyra Yezaensis) manufactured by Shiroko Co., Ltd.
  • the main components of the extract are an oligosaccharide of ex-1,3 linkage galactan sulfate and a galactan sulfate oligosaccharide consisting of ex1,3 linkage and ⁇ 1,4 linkage.
  • the method of application is selected to determine whether lung cancer (lung squamous cell carcinoma, lung adenocarcinoma, small cell lung cancer), thymoma, thyroid cancer, prostate cancer, renal cancer, bladder cancer, colon cancer , Rectal cancer, esophageal cancer, cecal cancer, ureteral cancer, breast cancer, cervical cancer, brain cancer, tongue cancer, pharyngeal cancer, nasal cavity cancer, larynx cancer, stomach cancer, liver cancer, bile duct cancer, testicular cancer, ovarian cancer It is effective in treating cortical cancer, metastatic bone cancer, malignant melanoma, osteosarcoma, malignant lymphoma, plasmacytoma, lip
  • the dose of the compound having a 1,3-glucan structure that is an NK activator or NKT activator is about lg-4 OgZ days, preferably about 5 g-20 gZ days
  • the compound having a ⁇ -1,3 glucan structure as an agent has an lg of about 10 gZ days, preferably about 3 g to 6 gZ days.
  • the administration period is generally 10 days to 24 months, and the administration frequency is every other day or 1 to 3 times Z days, preferably daily administration.
  • the cancer immunotherapy agent, CTL activator (IL-12 production inducer, INFy production inducer), NK activator, and NKT activator are preferably orally ingested.
  • TNF Q When an anticancer (chemotherapy) agent, radiation, or steroid combination therapy is performed in addition to the combination of the present invention, TNF Q; ⁇ IFN Y ⁇ IL-12 ⁇
  • the killer T cell lineage is severely impaired. Therefore, they are preferably not used in the present invention.
  • low-dose chemotherapy which is a method that does not impair the immune system, such as low-dose chemotherapy such as 5FU, UFT, mifurol, furturon, and CDDP ⁇ / zg-lO / zg
  • administration methods such as taxotere or low-concentration anticancer drugs such as taquinol, adriamycin, mitomycin, and CPT-11.
  • taxotere or low-concentration anticancer drugs such as taquinol, adriamycin, mitomycin, and CPT-11.
  • the measurement of NKT cells having NKR-P1 can be performed by measuring cell surface antigens (CD3 and CD161) specifically present on the cell surface of NKT cells. Specifically, for lymphocytes in peripheral blood, cells that are positive for CD3 and positive for CD161 [CD3 (+) CD161 (+)] are assayed. That is, CD3 and CD161 which are cell surface antigens of NKT cells are measured by a two-color test using a flow cytometry using a monoclonal antibody.
  • “activated NKT cells” means that the ratio of NKT [CD3 (+) CD161 (+)] cells in lymphocytes is 10% or more, more preferably 16% or more.
  • NKT cell activity is a function that increases the proportion of NKT cells to 10% or more, more preferably 16% or more, or a function that further enhances the proportion of NKT cells before administration of a certain substance.
  • CD3 (-) CD161 (+) refers to assaying cells that are negative for CD3 and positive for CD161. This method is useful for measuring NK cells.
  • CD8 (+) refers to assaying for CD8-positive cells. This method is useful for measuring CTL activity.
  • the blood cell of a cancer patient is used to detect cell surface antigens.
  • CD3, CD161, and CD8 were discriminated as positive or negative, and the proportion of each cell was measured by a two-color test using a flow cytometer as usual.
  • monoclonal antibodies against CD3, CD161 and CD8 were manufactured by Coulter or Betaton Dickinson, respectively.
  • lymphocytes in peripheral blood two of cell surface antigens, CD3, CD161, and CD8, and perforin are measured by a three-color test using flow cytometry as usual. Specifically, cells were fixed by collecting a fixation solution into the collected blood, and after adding a membrane permeate, the reaction was performed by adding an anti-perforin antibody (manufactured by Pharmingen) and reacting, followed by PRE-Cy5 labeling. A secondary antibody (manufactured by DAKO) was added and reacted, and then an anti-CD3-PE (Coulter 6604627) antibody and an anti-CD161-FITC (B-D) antibody were added and reacted. Measure by flow cytometry. Figures ⁇ Abbreviations in the table are indicated as P or PER. (Preparation of Sample for Measuring Site Force In)
  • a mononuclear cell fraction is separated and prepared from blood. Heparin-added peripheral blood is diluted 2-fold with Phosphate Buffered Saline (PBS) and mixed, then layered on FicoU-Conray solution (specific gravity 1.077), and 400 G for 20 minutes After centrifugation, the mononuclear cell fraction is collected. After washing, prepare RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), and adjust the cell number to 1 ⁇ 10 6.
  • PBS Phosphate Buffered Saline
  • FBS fetal bovine serum
  • Phytohemagglutinin manufactured by DIFCO was added to the obtained cell suspension 2001 to a concentration of gZml, and cultured in a 96-well microplate at 37 ° C for 24 hours in the presence of 5% CO. And in the cultured cell solution
  • a measurement kit by enzyme immunoassay (ELISA) available from R & D SYSTEMS or MBL which is a power that can use clinical and biochemical tests known per se.
  • ELISA enzyme immunoassay
  • MBL microplate
  • HRP horseradish peroxidase
  • the ability to induce IL-12 production refers to a function that enhances the amount of IL-12 produced by stimulation of the peripheral blood mononuclear cell fraction to 7.8 pgZml or more, or IL-12 production before administration of a certain substance.
  • a function that enhances the amount refers to a function that enhances the amount of IL-12 produced by stimulation of the peripheral blood mononuclear cell fraction to 7.8 pgZml or more, or IL-12 production before administration of a certain substance.
  • IFN y was measured by an enzyme immunoassay (EIA method) using an IFN y EASIA kit from BioSource Europe S. Actually, in each well of a 96-well microplate, dispense 50 1 of a standard solution or a 2-fold dilution of the sample prepared above, and dispense 50 ⁇ l of HRP-labeled anti-IFN- ⁇ antibody. The mixture was further reacted at room temperature with shaking for 2 hours. After removing the reaction solution from each well and washing three times, 200 1 of the chromogenic substrate solution was dispensed, and the mixture was allowed to react at room temperature for 15 minutes while shaking, and 50 1 of the enzyme reaction stopping solution was dispensed.
  • EIA method enzyme immunoassay
  • the Thl / Th2 ratio indicates the ratio of cells producing ⁇ (Thl) to cells producing IL-4 (Th2) among helper T cells having the cell surface antigen CD4, and CD4 (+ ) IFN y (+) / CD4 (+) IL-4 (+).
  • the Thl / Th2 cell ratio was determined by a helper T (Th) cell line Three Color analysis test by flow cytometry using a conventional method known to those skilled in the art, and specifically described in WO 02/04944. The test was carried out using the method described above.
  • Serum concentrations were measured by enzyme-linked immunosorbent assay (ELISA) (ACCUCYTE Human VEGF, ACCUCYTE Human bFGF, ACCUCYTE Human Endostatin: CYTIMMUNE Sciences Inc.) of the sales kit.
  • ELISA enzyme-linked immunosorbent assay
  • Each marker used in the clinical test was a commercially available marker, and the measured value was shown by each recommended method.
  • the displayed abbreviations are based on each general display method.
  • the response rate for each type of cancer indicates the proportion of CR, PR, LNC, SNC, and PD in all cases of each type of cancer.
  • MTC new immunotherapy
  • This NITC induces endogenous TNF ⁇ , IFN ⁇ , and IL-12 upon ⁇ -1,3 glucan administration to activate CTLs (killer T cells), and ⁇ and ⁇ upon ⁇ -1,3 glucan administration.
  • This is a BRM therapy that works to activate cells and also to inhibit angiogenesis by oral administration of Better Shear.
  • a cancer immunotherapeutic agent, an IL-12 production inducer, shark cartilage (Seishin Enterprise), and saccharides having an o1,3 structure were administered according to the recommended formulations.
  • ILX Tripi Pharmaceutical
  • ILY Seishin Pharmaceutical
  • Krestin Sankyo
  • etc. were administered alone or in combination as an IL-12 production inducer depending on the patient's condition.
  • the measurement of the amount of IL-10 in this example was performed by an ELISA method using a kit manufactured by Bio Source Europe. Actually, 50 1 of a standard solution or a sample prepared by the above-described method for preparing a site force-in sample was dispensed into each well of a 96-well microplate, and reacted at room temperature for 2 hours. After removing the reaction solution in each well, washing three times, dispensing 50 1 of peroxidase-labeled anti-human IL-10 monoclonal antibody, and reacting at room temperature for 2 hours, removing the reaction solution in each well, Washed three times.
  • the chromogenic substrate solution was dispensed at 200 1 each, and allowed to stand at room temperature for 25 minutes, and then the enzyme reaction stop solution was dispensed at 501 each.
  • the absorbance of each well at 450 nm was measured by Emax (manufactured by Wako Pure Chemical Industries, Ltd.).
  • the amount of IL-10 is the same as pgZml.
  • Iressa in patients with lung adenocarcinoma who have been effective after NITC
  • NKT8 (+) T cells [CD3 (+) CD161 (+) CD8 (+) cells] and Thl / Th2 ratio, IFNy, IL-12, and IL-10 as a negative factor (Fig. 1) ).
  • NKT (8+) is NKT8 (+ ⁇ CD3 (+) CD161 (+) CD8 (+)]
  • NKT (4+) is NKT4 (+) (CD3 (+) CD161 (+) CD4 (+)]
  • NKT8 (+) [CD3 (+) CD161 (+) CD8 (+)] cells NKT8 (+) P (+) [CD3 (+) CD161 (+) CD8 (+) perforin (+)] cells .
  • NKT4 (+) P (+) [CD3 (+) CD161 (+) CD4 (+) perforin (+)] cells there was no difference between effective and ineffective.
  • NKP (+) / NK CD3 (-) CD161 (+) perforin (+)
  • the ratio in the / CD3 (-) CD161 (+) cells increased only in the effective group (P ⁇ 0.05) (FIG. 7).
  • NKT8 (+) P (+)> CD8 (+) P (+)> NKP (+) is important, and Effector cells.
  • CD8 (+) P (+) (CD8 (+) perforin (+)) T cell NK P (+) (CD3 (-) CD161 (+) perforin (+)) cell effector effector in damaging cancer cells Forces that have become cells (Non-Patent Documents 3 & 4). It has been confirmed that it is a cell expressing the chemokine of CX3CR1, a perforin-granzyme B-positive cell that can exert strong cytotoxicity. These CX3CR1-expressing cells have been reported to be expressed in cells beyond a subset of NK cells and monocyte T cells (CD4, CD8, y ⁇ cells) and show strong cytotoxicity.
  • NKT8 (+) P (+) (CD3 (+) CD161 (+) CD8 (+) which is a cell induced by co-administration of NITC and Iressa and is at least a perforin-producing cell Perforin (+)] cells were the most potent.
  • cytotoxicity of CD8 (+) P (+) (CD8 (+) perforin (+)) T cells and NKP (+) (CD3 (-) CD161 (+) perforin (+)) cells in that order. Yes, but this cell is very similar to CX3CR1-expressing cells.
  • the marine yeast activates this group of cells, and the NITC or the marine yeast group is thought to activate CX3CR1-expressing cells.
  • IFN- ⁇ and IL-12 are not activated, but there are effector cells controlled by the Thl / Th2 ratio balance, and their cytotoxicity is CD8 (+ ) P (+) / CD8 (+) (CD8 (+) perforin (+) / CD8 (+)) T cells, NKT8 (+) P (+) / NKT8 (+) (CD3 (+) CD161 (+) CD8 (+) perforin (+) / CD3 (+) CD161 (+) CD8 (+)] cells, NK P (+) / NK (CD3 (-) CD161 (+) perforin (+) / CD3 (-) CD161 (+)] The activity is higher in the order of cells.
  • the CX3CR1-bearing cells induced by NITC of A. above have slightly lower cytotoxic potency than the Y cells (provisional name) similar to the CX3CR1-bearing cells induced by combined use of NITC and Iressa of B. it is conceivable that.
  • CD8 (+) P (+), NKT8 (+) P (+), and NKP (+) in order of decreasing cytotoxicity titer
  • CMOS complementary metal-oxide-semiconductor
  • Y cells cytotoxicity-activated cells
  • ⁇ cells are considered to be activated by an IL-21-mediated reaction (The Journal of Immunology, 2003, 171: 608-615).
  • CD4 (+) T cells Independent of cytoin and is not affected by CD4 (+) T cells.
  • CD8 (+) and NK cells which are hardly damaged by immune cells, show strong cytotoxicity, and are very similar to the force Y cells known to be mediated by IL-21R. .
  • Y cells may be activated and cultured with IL-21 and returned to the subject, or adjusted for rejection (for example, in the case of the same HLA) so that others can administer the Y cells.
  • the method for examining cell kinetics of the present invention it is possible to examine the efficacy of cancer treatment in a combination use of a tyrosine kinase inhibitor and a cancer immunotherapeutic agent, and more effectively. It is possible to perform cancer treatment. Further, by using the screening method of the present invention, a cancer immunotherapeutic agent having an excellent effect when used in combination with a tyrosine kinase inhibitor can be obtained, and the cancer immunotherapeutic agent can be used effectively for cancer treatment.

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Abstract

On cherche à améliorer l'efficacité d'un médicament thérapeutique de ciblage moléculaire et, pour ce faire, on utilise un moyen qui élève un taux de rémission complète, réduit la durée nécessaire à la rémission complète et produit des effets synergiques avec une immunothérapie. C'est-à-dire qu'on cherche à obtenir des effets synergiques par l'utilisation combinée d'une nouvelle immunothérapie centrée sur l'activité CTL, l'activité NKT, l'activité NK, VGEF et autres, avec un médicament thérapeutique de ciblage, notamment un inhibiteur de tyrosine kinase. On a remarqué qu'un facteur clé réside dans un médicament thérapeutique immunologique tel qu'un agent induisant la production de IL-12 qui affecte efficacement les cellules productrices de perforine. Cette invention concerne également une méthode d'examen d'une immunothérapie contre le cancer dans laquelle on utilise la cinétique des cellules productrices de perforine en tant que marqueur et un médicament thérapeutique contre le cancer qui est recherché par criblage avec la cinétique en tant que marqueur.
PCT/JP2004/017843 2003-12-02 2004-12-01 Méthode d'examen de la cinétique des cellules WO2005054496A1 (fr)

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JP2003403676 2003-12-02

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2390363A1 (fr) * 2006-06-02 2011-11-30 GlaxoSmithKline Biologicals s.a. Procédé d'identification si un patient est réactif ou non à l'immunothérapie basé sur l'expression différentielle du gène PRF1

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001082935A1 (fr) * 2000-04-28 2001-11-08 Orient Cancer Therapy Co.,Ltd. Remedes contre le cancer
WO2003039568A1 (fr) * 2001-11-06 2003-05-15 Orient Cancer Therary Co.,Ltd. Compositions anticancereuses
WO2004026341A1 (fr) * 2002-09-19 2004-04-01 Orient Cancer Therapy Co.,Ltd. Traitement du cancer par immunotherapie

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001082935A1 (fr) * 2000-04-28 2001-11-08 Orient Cancer Therapy Co.,Ltd. Remedes contre le cancer
WO2003039568A1 (fr) * 2001-11-06 2003-05-15 Orient Cancer Therary Co.,Ltd. Compositions anticancereuses
WO2004026341A1 (fr) * 2002-09-19 2004-04-01 Orient Cancer Therapy Co.,Ltd. Traitement du cancer par immunotherapie

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
HERBST R.S. ET AL: "Targeting the epidermal growth factor receptor in non-small cell lung cancer", CLIN. CANCER RES., vol. 9, 1 December 2003 (2003-12-01), pages 5813 - 5824, XP002988021 *
MARUYAMA S. ET AL: "Tangan Mouse ni Okeru Gefitinib to Men'eki Ryoho Heiyo ni Tsuite no Igi", JAPANESE JOURNAL OF CANCER AND CHEMOTHERAPY, vol. 30, no. 11, October 2003 (2003-10-01), pages 1773 - 1775, XP002984971 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2390363A1 (fr) * 2006-06-02 2011-11-30 GlaxoSmithKline Biologicals s.a. Procédé d'identification si un patient est réactif ou non à l'immunothérapie basé sur l'expression différentielle du gène PRF1

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