WO2004087208A1 - 熱ショックタンパク質と磁性微粒子からなる悪性腫瘍の温熱治療剤 - Google Patents
熱ショックタンパク質と磁性微粒子からなる悪性腫瘍の温熱治療剤 Download PDFInfo
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- WO2004087208A1 WO2004087208A1 PCT/JP2003/003825 JP0303825W WO2004087208A1 WO 2004087208 A1 WO2004087208 A1 WO 2004087208A1 JP 0303825 W JP0303825 W JP 0303825W WO 2004087208 A1 WO2004087208 A1 WO 2004087208A1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0023—Agression treatment or altering
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
Definitions
- Hyperthermia for malignant tumors composed of heat shock proteins and magnetic particles
- the present invention relates to a hyperthermia for malignant tumors, and more particularly, to a hyperthermia for malignant tumors containing a heat shock protein and magnetic fine particles.
- malignant tumors still account for more than 30% of deaths, and there is a need to develop new treatments for malignant tumors. Therefore, the development of new therapeutic methods such as gene therapy, immunotherapy, and hyperthermia has begun.
- hyperthermia is an old treatment that has been practiced since ancient Greece, and uses the property that malignant tumor cells are more vulnerable to heat than normal cells.
- a widely used method of hyperthermia is to heat the entire site where the malignant tissue is located, to weaken the heat, and to kill the malignant tumor cells.
- a method is known in which magnetic fine particles are used as an internal heating element, and the magnetic fine particles are heated by electromagnetic waves.
- magnetite is coated with a positively charged lipid membrane (liposome) to form magnetite cationic liposomes (MCL) and use this (Shinkai et al.) , Jpn. J,
- Hyperthermia for malignant tumors has attracted attention because it is a non-invasive treatment, but this hyperthermia alone can completely cure various malignancies. Is often difficult.
- the present invention has been achieved by finding that the use of a heat shock protein significantly improves the therapeutic effect of hyperthermia for malignant tumors, particularly for hyperthermia of malignant tumors using magnetic fine particles. is there.
- the present invention is a hyperthermic therapeutic agent for malignant tumors, comprising a heat shock protein and magnetic fine particles.
- the present invention is further a hyperthermia therapeutic agent for malignant tumor, comprising a vector into which a heat shock protein gene is incorporated so that the heat shock protein can be expressed in malignant tumor cells, and magnetic fine particles.
- thermotherapeutic agent of the present invention may contain the heat shock protein and the magnetic fine particles, or the vector into which the heat shock protein gene is incorporated and the magnetic fine particles together or separately.
- the heat shock protein used in the present invention is not particularly limited as long as it exerts the effects of the present invention.
- the method for producing the heat shock protein used in the present invention is not particularly limited, and the heat shock protein includes naturally occurring heat shock proteins, those obtained by genetic recombination, and those obtained by chemical synthesis. .
- any material can be used as long as it absorbs electromagnetic waves and generates heat and is harmless to the human body, but it can generate heat by absorbing electromagnetic waves having a frequency that is hardly collected by the human body.
- ferromagnetic particles It can be preferably used because of its good absorption efficiency. Examples thereof include ceramics such as magnetite and ferrite, and ferromagnetic metals such as permalloy.
- the magnetic fine particles preferably have a particle size of 5 or less, particularly 1 ⁇ or less.
- MCL magnetite cationic ribosomes
- the magnetic fine particles preferably used in the present invention are magnetic fine particles having a surface to which an antibody that selectively binds to malignant tumor cells is bound.
- the magnetic microparticles to which the antibody is bound are selectively concentrated near the malignant tumor cells, so that hyperthermia can be performed without heating other than the malignant tumor cells.
- a vector having a heat shock protein gene incorporated therein so that the heat shock protein used in the therapeutic agent of the present invention can be expressed in malignant tumor cells can be obtained by adding a heat shock protein gene together with an appropriate regulatory gene together with plasmid ⁇ virus or the like.
- the vector can be obtained by integrating the vector so that it can be expressed.
- the present invention also relates to the use of heat shock proteins in hyperthermia for malignant tumors. That is, the present invention provides a method for hyperthermia treatment of a malignant tumor, which comprises administering a heat shock protein to a malignant tumor and then thermally treating the malignant tumor. And administering the malignant tumor to an electromagnetic field after administration to the patient.
- the heat shock protein and the magnetic fine particles can be administered to a malignant tumor at the same time or at a later time, but it is preferable to administer them simultaneously.
- the heat shock protein and the magnetic microparticles are desirably administered to the malignant tumor tissue and its vicinity.
- the invention further relates to the use of the heat shock protein gene in hyperthermia treatment of malignant tumors. That is, a vector incorporating a heat shock gene so that a heat shock protein can be expressed in malignant tumor cells is injected into the malignant tumor, A method for hyperthermia treatment of malignant tumors, which comprises expressing a heat shock protein in a malignant tumor, followed by hyperthermia treatment of the malignant tumor. Injecting a vector incorporating a shock protein gene into a malignant tumor to express the heat shock protein in the malignant tumor cells, and then administering the magnetic microparticles to the malignant tumor and then placing the malignant tumor in an electromagnetic field The heat treatment of malignant tumors. The administration of the magnetic fine particles to the malignant tumor is preferably performed after the heat shock protein is sufficiently expressed in the malignant tumor cells.
- the malignant tumor according to the present invention may include all kinds of malignant tumors.
- skin cancer such as malignant melanoma, lung cancer, colon cancer, breast cancer, brain tumor, malignant histiocytoma, osteosarcoma, liver cancer
- examples include prostate cancer, spleen cancer, esophageal cancer, bladder cancer, lung cancer, ovarian cancer, uterine cancer, and stomach cancer, particularly malignant melanoma, liver cancer, prostate cancer, and colon cancer.
- a high-frequency magnetic field is preferably used, and in particular, a high-frequency magnetic field generated by an electromagnetic wave having a frequency of 1 to 10 MHz is preferable. ⁇ ⁇
- the reason why a high frequency magnetic field with a higher frequency is preferable is that magnetic hysteresis heating efficiency is high, and the reason why a high frequency magnetic field with a frequency lower than 1 O MHz is preferable is that heat generation of a living body due to an induced current is caused. This is because it is possible to heat the magnetic fine particles without heating.
- FIG. 1 shows the flow of an experiment of hyperthermic immunotherapy using rmHSP70 described in Example 1.
- FIG. 2 shows the results of the IFN- ⁇ -producing ability determined by ELISSOT in Example 1.
- the ability of the spleen cell population after ex vivo treatment to secrete IFN- ⁇ was evaluated using the ELISPOT assay.
- FIG. 3 shows the temperature changes in the tumor ( ⁇ ) and rectum (hata) during magnetic field irradiation in Example 1. Data points and bars indicate the mean and SD of 5 mice.
- FIG. 4 shows the change in tumor volume due to the combination of administration of rmHSP70 and hyperthermia in Example 1. After MCL injection, no heat treatment; B Group:: A group MCL after injection, there heat treatment; C Group: rmHSP 70 a post-dose 80 mu [delta], no heat treatment; D Group: MCL and 20 / zg of r mH SP After administration of 70, there was a heat treatment; Group E: MCL and 80 / g of rmHSP70, after the heat treatment.
- FIG. 5 shows the survival rate of the tumor-bearing mice 90 days after the hyperthermia treatment in Example 1.
- Group A (n 10): Garden;
- Group B (n 10): Mouth;
- Group C (n 10): #;
- Group D (n 10): ⁇ ;
- Group E (n 10) : ⁇ .
- FIG. 6 shows the cytotoxic activity of spleen cells against B16 melanoma cells in Example 1.
- Spleen cells were from mice in group E ( ⁇ ) and naive mice ( ⁇ ) two weeks after the heat treatment.
- the effector: target (E: T) ratio was 100: 1 to 25: 1.
- Data points and plots show the mean, SD, of three independent experiments.
- FIG. 7 shows the structure of the plasmid used in Example 2.
- FIG. 8 shows the flow of an experiment of hyperthermia using the hsp70 gene in Example 2.
- FIG. 9 shows the antitumor effect of Example 2 by the combination of administration of the hsp70 gene and hyperthermia.
- no hyperthermia treatment (F); after administration of MCL, hyperthermia treatment: (G); hsp70 administration: (H); after administration of nu11 and MCL, hyperthermia treatment: (I); Heat treatment after administration of hsp 70 and MCL (J).
- FIG. 10 shows the survival rate of the tumor-bearing mice 90 days after the hyperthermia treatment in Example 2.
- FIG. 11 shows the cytotoxic activity of spleen cells against B16 melanoma cells in Example 2.
- Spleen cells were from mice in group J ( ⁇ ) and na ⁇ ⁇ ve mice (violent) two weeks after the heat treatment.
- the effector: target (E: T) ratio was 100: 1 to 25: 1.
- Data points and data represent the mean SD of three independent experiments.
- FIG. 12 shows the concentration of inducible HSP 70 24 hours after in vivo hyperthermia treatment in Example 2. Inducible HSP70 concentrations in tumors were determined by ELISA. No treatment group (white bar); heat group (gray bar); rmH SP 70 (80 ⁇ g) + heat group (dot bar); hsp 70 gene + heat group (black bar).
- the magnetic fine particles having an antibody that selectively binds to malignant tumor cells on the surface thereof used in the present invention can be prepared by, for example, the method described in Japanese Patent Application Laid-Open No. 3-128331,
- the microparticles can be produced by binding a bifunctional crosslinking agent to the microparticles and then reacting the microparticles with an antibody that selectively binds to malignant tumor cells.
- Examples of the antibody that selectively binds to the malignant tumor cell include a monoclonal antibody against lung cancer (HB4C5), a monoclonal antibody against colon cancer (17-1A), and a monoclonal antibody against breast cancer (H1 5 F 2) etc. may be used. .
- the magnetic fine particles are a ferromagnetic metal
- Examples of a method of binding the bifunctional crosslinking agent include a method of sequentially binding ⁇ -aminoprovir triethoxysilane and dartalaldehyde to the magnetic fine particles, and a method of binding buraldehyde and acrylaldehyde in order.
- the heat shock protein used in the present invention is a molecular chaperone protein that exists in a wide range of prokaryotes and eukaryotes and accounts for about 5% of the total intracellular protein. As the name implies, heat shock proteins are highly expressed by heat and other stresses, and their share in intracellular proteins increases to 15%. The functions of heat shock proteins, which are expressed in many cells, are diverse.
- mouse melanoma B16 melanoma cells As malignant tumor cells, mouse melanoma B16 melanoma cells (Riken Cell Bank) were used. The cells were cultured in DMEM medium (Gibco BRL) containing 10% calf serum, antibiotics (100 U / ml sodium penicillin G, 0.1 g / ml streptomycin sulfate) at 37 ° (: 5% CO 2 And 95% air in a carbon dioxide incubator.
- DMEM medium Gibco BRL
- antibiotics 100 U / ml sodium penicillin G, 0.1 g / ml streptomycin sulfate
- mice were C57B1 / 6 female, 4 weeks old (Charles River Japan) 0
- B 16 melanoma cells were suspended in 50 ⁇ l of phosphate buffer (PBS, 0.05% sodium phosphate, 0.15M sodium chloride).
- PBS 0.05% sodium phosphate, 0.15M sodium chloride
- the cell suspension was implanted into the skin of the right thigh of the mouse using a 29 G syringe (Becton Dickinson).
- Magnetite manufactured by Toda Kogyo
- a particle size of 1 Onra was used as the magnetic fine particles.
- the magnetite was thoroughly washed with water to remove excess ion components, and subjected to ultrasonic treatment to obtain a magnetite colloid solution dispersed in water.
- TMAG N— ( ⁇ -trimethylammonioacetyl) -didodecyl-D-glutamate chloride) (manufactured by Mutual Pharmaceutical Co., Ltd.), DLPC (dilauroylphosphatidylcholine) (manufactured by Sigma), DOPE (geoleyl) Phosphatidylethanol / reamine) (Sigma) was dissolved in a chromatographic flask at a molar ratio of 1: 2: 2 (TMAG: DLPC: DOPE), and the solution was placed in an eggplant-shaped flask. The solvent was removed with a rotary evaporator. A phospholipid membrane was formed on the inner wall of the flask.
- recombinant mouse HSP70 As a protein, recombinant mouse HSP70 (rmHSP70, manufactured by BioDynamics Laboratory) was used.
- a transistor inverter As a device for irradiating a magnetic field, a transistor inverter (LGH-100-05; manufactured by Dai-ichi Kogyo Kogyo) and a vertical coil (inner diameter 7 cm, length 7 cm) were used.
- the magnetic field irradiation was performed such that the cell mass was at the center of the coil.
- the temperature was measured using an optical fiber thermometer (FX-9030; Anritsu meter) that was not affected by the alternating magnetic field.
- the r mH SP 70 group cells were added HS P 7 0 protein dissolved in PBS (8 0 g / l XI 0 7 cells). The control group received the same amount of PBS.
- the output of the magnetic field irradiation device was adjusted and the temperature of the cell mass was maintained at 43 ° C.
- the magnetic field treatment was for 30 minutes.
- mice After completion of magnetic field irradiation, they were transplanted B 1 6 melanoma cells 1 X 1 0 7 pieces of mice subcutaneously with upper cleansed.
- B16 melanoma cells were heated ex vivo, and the IFN- ⁇ -producing ability of the spleen cells of the transplanted mice was measured by the ELIS POT method.
- Mouse Interferon- ⁇ ELIS POT MABTECH was used for the measurement.
- Anti-IFN-antibody i (AN18; 15 ⁇ g / ml) was coated on a hydrophilic mixed cellulose ester membrane-attached plate (manufactured by Millipore) in an amount of 100 ⁇ l each and coated. It was left overnight at 4 ° C. Washed with PBS and 5% 83 in? Blocked at 83 (2 hours, 37 ° C). After blocking was completed, mouse spleen cells (1.0 X 10 5 cells) were introduced. Spleen cells were removed from mice 2 weeks after transplantation Then, cells were separated using Mediamachine System (DAKO A / S).
- DAKO A / S Mediamachine System
- the cells were hemolyzed using 0.75% NH 4 C 1 and suspended in an RPMI medium (manufactured by Gibco) to obtain a cell suspension (5.0 ⁇ 10 5 cells / ml). This was cultured at 37 for 40 hours. After washing with PBS, anti-IFN- ⁇ antibody ii (R4-6 ⁇ 2; 1 ⁇ g / ml) was added at 100 ⁇ 1 each and cultured for 2 hours at room temperature. After washing with PBS, 100-fold diluted streptavidin monoalkaline phosphatase (100-fold) was added, and the cells were cultured at room temperature for 1 hour.
- the substrate (BCI I / II; Moss) was added in an amount of 10 ⁇ , and the mixture was incubated for 1 hour. When spots were confirmed, tap water was added to stop the reaction. After the plate was dried, the number of spots was counted using a stereomicroscope (SZH10; manufactured by Olympus).
- MCL (2 Omg / rnl) was injected 0.1 ml into the tumor.
- MCL injection was directly injected into the tumor using a 26 G syringe (manufactured by Terumo Corporation). The injection speed was 0.2 ml / h.
- the device for irradiating the magnetic field used a transistor inverter (LGH-100-05; manufactured by Dai-ichi Kogyo Kogyo) and a horizontal coil (inner diameter 7 cm, length 7 cm).
- the magnetic field irradiation was such that the tumor part of the mouse under anesthesia was in the center of the coil.
- the temperature of the tumor surface and rectum during magnetic field irradiation was measured using an optical fiber thermometer (FX-9030; manufactured by Anritsu Keiki) which was not affected by the alternating magnetic field.
- FX-9030 manufactured by Anritsu Keiki
- recombinant mouse HS P70 As a protein, recombinant mouse HS P70 (rmHS P70, manufactured by Bio Dynamics Laboratory) was used. The protein for one mouse was dissolved in 201 PBS, mixed with 0.1 ml of MCL (20 mg / ml), and injected directly into the tumor using a 26 G syringe. The injection speed was 0.24 ml / h.
- Group A received only MCL and was not irradiated with a high-frequency magnetic field (no treatment group).
- group B the high frequency field after MCL administration was irradiated (warm heat group).
- group C rmHS P70 protein was administered at a rate of 80 / ig per animal (HSP-administered group).
- Group D received 20 / zg of rmHSP70 protein and MCL per animal, and was irradiated with a high-frequency magnetic field (HSP (20 / ig) + thermal group).
- Tumor volume was measured every three days from the start of treatment to evaluate the treatment effect. Tumor volume (cm 3 )
- mice spleen cells of the HSP 80 ⁇ ⁇ + heat and heat combined group as effector cells.
- the spleen of a naive mouse (C57Bl / 6, 7-week-old female) was used as a control.
- Cells were separated from spleen using Medimachine System (DAKO A / S). Thereafter, the cells were hemolyzed using 0.75% NH 4 C 1 and suspended in an RPMI medium (manufactured by Gibco) to obtain a cell suspension.
- B 1 6 melanoma cells IX 1 0 6 cells / ml
- Effector cells 50 / il
- target cells 50 / il
- the tumor volume was compared 30 days after the start of treatment, and the significance was evaluated using the Mann-Whitney U test.
- the significance of the survival rate was evaluated by the log rank test of the Kaplan-Meier curve.
- the ELISPOT method is a technique that stains antibody molecules and rhinoceros-in produced by immune cells and detects them at the cell level.
- IFN_ ⁇ one of the cytokines, as an indicator of CTL (cytotoxic T lymphocytes). Therefore, an experiment was performed to determine whether the immunity was activated by measuring the IFN- ⁇ -producing ability of the lymphocytes in the spleen using the ELIS POT method.
- FIG. 2 shows the number of spots per 1 ⁇ 10 5 spleen cells.
- FIG. 3 shows the temperature changes on the tumor surface and in the rectum during magnetic field irradiation.
- the temperature of the tumor surface reached 43 ° C two minutes after the start of the magnetic field irradiation.
- the tumor could then be heated at exactly 43 ° C for 30 minutes by manipulating the output of the device. Turning off the instrument output reduced the tumor surface temperature to 38 ° C.
- the temperature in the rectum hardly increased during magnetic field irradiation, and remained at a normal value.
- Figure 4 shows the change in tumor volume from the start of treatment. Tumor volume continued to increase in the untreated group.
- the heat-treated group the HSP-administered group, and the HSP (20, ug) + heat-treated group
- the tumor growth was suppressed for about one week, but the tumor volume continued to increase after one week, and the tumor volume after 30 days Did not differ significantly from the untreated group.
- Figure 5 shows the change in survival rate from the start of treatment.
- the heat treatment group, the HSP treatment group, and the HSP (20, ug) + heat treatment group all died by 50 words.
- the two mice whose tumors were completely cured survived even after 90 days, and there was a significant difference in the survival rate between the HSP (80 g) + heat-treated group and the untreated group and the heat-treated group. Prolonged survival was seen depending on the dose of HSP70 protein as well as tumor volume.
- mice melanoma B16 melanoma cells (Riken Cell Bank) were used. The cultivation was performed in the same manner as in Example 1-1. The mice used were C57B1 / 6 female, 4 weeks old (Charles River Japan).
- Hsp70 has a human inducible hsp7 OcDNA downstream of the CMV promoter.
- pCMVhygro is Xbal, it was obtained by removing hs ⁇ 7 OcDNA by blunt E command ligated with Kpnl site ( Figure 7) 0
- Gene transfer was performed by the cationic ribosome method. Using a lipid having the same composition as the MCL, the solvent was removed in the conical tube to prepare a lipid membrane. To this, PBS and 20 ⁇ g of plasmid were added and vortexed to prepare a gene suspension to a final concentration of 0.2 mg / ml. The gene suspension was directly injected into the tumor at a dose of 20 g per mouse from 5 different places at 5 sites. The injection speed was 0.2 ml / h. -1-3 MCL administration and high frequency magnetic field irradiation
- MCL was produced in the same manner as in 1-3 of Example 1.
- the transgenic group received MCL one day after transfection.
- the MCL administration method and high-frequency magnetic field irradiation were performed in the same manner as in 1-4 of Example 1.
- group F After tumor tissue transplantation, the experimental animals were divided into five groups. In group F, only the MCL was administered and the high-frequency magnetic field was not irradiated (no treatment group). Group G was irradiated with a high-frequency magnetic field after MCL administration (thermal group). Group H received pCMVhygro.hsp70 (hsp group). Group I was treated with pCMVhygro and hyperthermic treatment with MCL (nul 1
- plasmid pCMVhygro.hsp70 was transfected by the method described in 1-2 above. Twenty-four hours after transfection, blood was removed from the heart by perfusion of PBS and the tumor was removed. Immediately after removal, the tumors were placed in 0. C. T. Compound (Sakura Seiki) and frozen in liquid nitrogen. Tissues were sliced to a thickness of 4111 and fixed with acetone for 5 minutes. The frozen sections were cultured at 37 ° C for 60 minutes in an anti-HSP70 antibody (clone: K-20, Santa Cruz Biotechnology) that specifically binds to inducible HSP70.
- an anti-HSP70 antibody clone: K-20, Santa Cruz Biotechnology
- the cytotoxic activity was measured using mouse spleen cells of the hsp + heat group as effector cells.
- the measurement method and the calculation method of the cytotoxic activity were performed in the same manner as in Example 1, 117.
- the amount of inducible HSP expression in the tumors of the mice in the untreated group, the heat group, the HSP (80 ⁇ g) + heat group, and the hsp + heat group was measured.
- the measurement was performed using Hsp70 ELISA Kit (StressGen Biotechnologies). Twenty-four hours after irradiation with a magnetic field, blood was removed from the heart by perfusion of PBS, and the tumor was excised.
- the tumor volumes 30 days after the start of treatment were compared, and the significance was evaluated using the Mann-Whitney U test.
- the significance of the survival rate was evaluated by the log-rank test of the Kaplan-Meier curve.
- Inducible HS P70 protein in tumors 24 hours after gene transfer Detected by FITC. No fluorescence was observed in the tumor without treatment with the null1 gene, and no inducible HSP70 was expressed.
- a magnetic field was irradiated immediately after MCL administration.
- the temperature changes in the tumor surface and in the rectum during the magnetic field irradiation were the same as in the case of using the HSP70 protein in Example 1 in combination with hyperthermia. It was possible to heat only the cancerous tissue without overheating the normal tissue.
- FIG. 9 shows the change in tumor volume from the start of treatment.
- Tumor volume in the untreated group continued to increase.
- the tumor growth was also temporarily suppressed in the heat group, hsp group, and nu 11 + heat group, but the tumor volume continued to increase overall, and the tumor volume after 30 days was significantly higher than that in the untreated group. No difference was made.
- the change in survival rate from the start of treatment is shown in FIG.
- the mice in the untreated group, the heat group, the hsp group, and the nu11 + heat group all died by 50 days later.
- 50% of the mice in the hsp + heat group survived even after 50 days.
- the three animals whose tumors were completely cured were still alive 90 days later.
- the hsp + heat group had a significant difference in survival rate compared to the other four groups.
- mice in the hsp + hyperthermic group had acquired antitumor immunity.
- the spleen cells in the hsp + heat group showed higher cytotoxic activity than naive mice. From these results, it was found that antitumor immunity against B16 cells was activated by the combination of gene therapy with the hsp70 gene and hyperthermia.
- the expression level of the inducible HSP70 protein was measured 24 hours after the heat treatment (FIG. 12).
- the hyperthermic therapeutic agent for malignant tumors of the present invention has a higher therapeutic effect on various malignant tumors than conventional hyperthermic therapeutic agents.
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AU2003227265A AU2003227265A1 (en) | 2003-03-27 | 2003-03-27 | Thermotherapeutic for malignant tumor comprising heat shock protein and fine magnetic particles |
JP2004570122A JPWO2004087208A1 (ja) | 2003-03-27 | 2003-03-27 | 熱ショックタンパク質と磁性微粒子からなる悪性腫瘍の温熱治療剤 |
PCT/JP2003/003825 WO2004087208A1 (ja) | 2003-03-27 | 2003-03-27 | 熱ショックタンパク質と磁性微粒子からなる悪性腫瘍の温熱治療剤 |
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2006273740A (ja) * | 2005-03-29 | 2006-10-12 | Takeshi Kobayashi | 抗hmw−maa抗体を結合した磁性微粒子を含むリポソーム |
WO2008017507A2 (en) * | 2006-08-11 | 2008-02-14 | Roche Diagnostics Gmbh | Nanoparticle nucleic acid bonding compound conjugates forming i-motifs |
CN109148067A (zh) * | 2018-07-16 | 2019-01-04 | 中国计量科学研究院 | 表面共价有机框架材料修饰的磁性纳米材料及制备、应用 |
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WO1994011513A1 (en) * | 1992-11-13 | 1994-05-26 | Medical Research Council | Heat shock proteins and the treatment of tumours |
WO1999049881A2 (de) * | 1998-03-27 | 1999-10-07 | Gabriele Multhoff | Verwendung von hsp70 protein |
EP0969100A1 (en) * | 1997-09-29 | 2000-01-05 | HSP Research Institute, Inc. | Method for screening heat shock protein expression induction regulators |
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2003
- 2003-03-27 WO PCT/JP2003/003825 patent/WO2004087208A1/ja active Application Filing
- 2003-03-27 JP JP2004570122A patent/JPWO2004087208A1/ja active Pending
- 2003-03-27 AU AU2003227265A patent/AU2003227265A1/en not_active Abandoned
Patent Citations (3)
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CN109148067A (zh) * | 2018-07-16 | 2019-01-04 | 中国计量科学研究院 | 表面共价有机框架材料修饰的磁性纳米材料及制备、应用 |
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AU2003227265A1 (en) | 2004-10-25 |
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