JP2002069005A - Solid carcinoma-curing formulation - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a solid carcinoma-curing formulation capable of effectively killing carcinoma cells without adverse effects. SOLUTION: This solid carcinoma (e.g. a uterine cervix cancer, a prostatic cancer, a mammary cancer)-curing formulation contains arsenious acid (As2O3) and an ASK1-SEK1-JNK cascade-activating material. Especially, an ASK1- activating material is the material capable of dissociating thioredoxin from ASK1 and L-buthionine sulfoximine (LBSO) is preferable. Here, ASK1 is the abbreviation of apotosis signal-regulating kinase 1, SEK1 is that of SAPK(stress- activated protein kinase)/ERK(extracellular signal-regulated protein kinase) and JNK is that of c-Jun NH2-terminal kinase.

Description

DETAILED DESCRIPTION OF THE INVENTION

TECHNICAL FIELD The invention of this application relates to a preparation for treating cancer. More specifically, the invention of this application relates to a solid cancer therapeutic preparation that effectively kills cancer cells without side effects and with little burden on patients.

2. Description of the Related Art In 1990, about 5.2 million people died from cancer worldwide, but in 2010,
It is estimated to approach 8 million (Pisani, P., Pa
rkin, DM, Bray, F., Ferlay, J., Int. J. Cancer 8
3, 18-29 (1999)), and the number of cancer deaths in Japan in 1999 exceeded 290,000 (Ministry of Health and Welfare 1999 demographic statistics). The ratio by site is high for stomach cancer and lung cancer, followed by liver cancer. Cancer control is regarded as one of the most important issues in disease control worldwide, and research on therapeutic agents and treatments is being conducted from various angles. To date, cancer has been treated by chemotherapy to kill cancer cells by administration of anticancer drugs, surgical treatment to remove the affected area, radiation therapy to irradiate the affected area, and normal white blood cell type matching. Hematopoietic stem cell transplantation (bone marrow transplantation) or the like is generally performed, in which a healthy hematopoietic stem cell is provided and the bone marrow affected by leukemia is replaced with normal bone marrow. Antimetabolites such as fluorouracil, alkylating agents such as cyclophosphamide, antibiotics such as daunorubicin hydrochloride and bleomycin, and anticancer drugs such as plant alkaloids used in chemotherapy all have the same therapeutic effect as nausea and vomiting. It is accompanied by strong side effects such as fever, decreased white blood cells and platelets, hair loss, and stomatitis. Also,
Surgery is effective at an early stage when the cancer is limited to a specific site, but is less effective when the cancer has advanced and metastasized. Radiation therapy is likewise a topical therapy and is not suitable for systemic cancer. Hematopoietic stem cell transplantation requires the provision of normal hematopoietic stem cells from a leukocyte-matched stranger, with a matching rate of 25% in siblings and 1% in parents and relatives.
% Or less, and also imposes a burden such as general anesthesia on the provider. As described above, all of the conventional cancer treatment methods have substantially the same or better effects on the patient. Recently, it has been reported that t (15; 17) positive acute promyelocytic leukemia (APL) cells are killed by arsenite (As ₂ O ₃ ) (Che
n, GQ et al., Nat.Genet. 20, 259-265 (1998); Zh
u, J. et al., Proc. Natl. Acad. Sci. USA 94, 39
78-3983 (1997)) as a new cancer treatment. According to these reports, As ₂ O ₃ is converted to PML-RARa (retinoic acid receptor (R
AR) causes degradation of α and PML (promyelocytic leukemia cell) fusion protein, causes remodeling of PMLbody which is an oncogenic domain of PML, and accumulation of its aggregates causes APL cell death . In addition, it has been found that such APL cell death occurs sufficiently with 1 μM or less of As ₂ O ₃ . On the other hand, for solid cancer,
As mentioned above, more than 90% of cancer deaths are due to solid cancer (Ministry of Health and Welfare 1999 demographic statistics)
In order to kill cancer cells with As ₂ O ₃ , it is necessary to administer a large amount (several μM or more), which involves a danger of toxicity and the like, so that the effect of cancer treatment expected from APL cannot be expected. It is the fact that it was being done. Therefore, the invention of this application has been made in view of the circumstances as described above, and solves the problems of the prior art, has no toxicity or side effects, and can effectively treat solid cancer with a small burden on patients. An object of the present invention is to provide a solid cancer therapeutic preparation that can be treated.

Means for Solving the Problems The present inventors have
In addition, at least eight neurodegenerative diseases (Huntington
Disease, Machado-Joseph disease, etc.)
Polyglutamine, thought to be a molecule, is at the core of PMLbody
Accumulated and SEK1-JNK (SAPK (stress-activa
tedproteinkinase) ／ERK (extracellular signal-r
egulated proteinkinase)Kinase-c-Jun
NH_Two-TerminalKinase) Cell death kinase
PC12 nerve cultured to activate zeta cascade
Confirmed and reported from tests on cells (Yasuda,
S. et al., Genes Cells 4, 743-756 (1999)). Soshi
Thus, the present inventors_TwoO_ThreeOf APL cell death
Phenomena observed in such PC12 neurons
Is very similar to that of cell death caused by polyglutamine
And attention. In addition, the inventors have proposed polyglutamine.
Activated in cell death of neurons
As accumulated in the MLbody, As_TwoO_ThreeA with addition of
PL cells, especially NB4 cells (Lanotte, M. et al., Bl
ood 77, 1080-1076 (1997)).
Accumulation of activated SEK1 was observed in
(See Reference Examples 1 to 3). From these findings, APL cells
In, As_TwoO_ThreeOf PMLbody caused by
Is considered to be causing cell death.
Activating substance that causes or promotes composition
Coexist in cancer cells to kill cancer cells
Thought that you can. And the inventors are dedicated to
The role of PMLbody accumulation in cancer cells
After clarifying the essentials, the present invention was completed. This application
The invention of the above was made according to the circumstances described above.
As a solution to the problems of the prior art,
Is ASK1-SEK1-JNK other than arsenous acid
Solid cancer treatment characterized by containing a cascade activator
A therapeutic formulation is provided. Second, the invention of this application
The above-mentioned solid cancer treatment, wherein the concentration of arsenate is 0.1 to 2 μM.
A formulation is provided. The invention of this application relates to any of the aforementioned
Thirdly, in the preparation for treating solid cancer, A other than arsenite is used.
SK1-SEK1-JNK cascade activator is JN
Fourth, AS activators other than arsenous acid
K1-SEK1-JNK cascade activator is SEK
Fifth, AS other than arsenous acid
K1-SEK1-JNK cascade activator is ASK
(1) Activating substance is provided as a preferred embodiment thereof.
I do. Sixth, the invention of this application is based on the fifth aspect.
In the invention of the above, the ASK1 activator is
Substance that causes the dissociation of thioredoxin,
7 shows such a solution of thioredoxin from ASK1.
The substance that causes separation is an active oxygen generating substance.
The present invention provides a preparation for treating solid cancer, which is characterized in that: And this out
Eighth, the invention of the application is the fifth invention, wherein the AS
K1 activator is L-buthionine sulfoximine
Also provided are formulations for treating solid cancer.

DETAILED DESCRIPTION OF THE INVENTION
The drug is a cancer cell revealed by the inventors' intensive studies.
Treats solid tumors based on the mechanism of cell death
You. That is, the solid cancer therapeutic preparation of the invention of the present application comprises A
By SK1-SEK1-JNK cascade activator
Activated ASK1-SEK1-JNK cascade
Causes death of cancer cells
This mechanism is applied. As mentioned above, As_TwoO_Three
Has a low concentration of 1 μM against leukemia cells.
It has been known to induce spore death.
For cancer, As_TwoO_ThreeNeed to be added at high concentration
Was considered. In the invention of this application, the inventors
Study reveals the mechanism of cancer cell death
As a result, ASK1-SEK1-JNK cascade activation
Activating substance As_TwoO_ThreeLow concentration of A
s_TwoO_ThreeBut it can effectively kill solid tumors
Formulations for treating form cancer are provided. Therefore, this application
The pharmaceutical preparation for treating solid cancer according to the invention comprises at least arsenous acid (As
_TwoO_Three) And ASK1-SEK1-JNK residue other than arsenous acid
What is necessary is just to contain a kade activator,
The shape, the method of ingestion and the like are not particularly limited. Such a fixed
In the preparation for the treatment of morphological cancer, As_TwoO_ThreeThe concentration of
Preferably it is 2 μM. More preferably, it is 0.
5-1 μM. As_TwoO_ThreeLess than 0.1μM
In some cases, cell death of cancer cells does not occur.
If it is higher than the normal
Because it also destroys the patient and causes pain to the patient.
Absent. In the preparation for treating solid cancer of the invention of the present application, A
SK1-SEK1-JNK cascade activator
May be used, and is not particularly limited. An example
For example, JNK (c-JunNH_Two-terminalkinase) activator
Quality, SEK1 (SAPK (stress-activatedproteinkinas
e) /ERK (extracellular signal-regulated proteink
inase)kinase) activator or ASK1 (apopt
osissignal-regulatingkinase 1) Activator etc.
No. JNK activators include SEK1 activity
Activators and ASK1 activators have been considered.
A substance that suppresses the action of the K activation inhibitor
Is also good. Further, as the SEK1 activator, for example,
Hydrogen peroxide (H_TwoO_Two), But SEK1 activity
Various substances that inhibit the action of
Is also good. Further, ASK1 activators include ASK1
A substance that causes the dissociation of thioredoxin from 1
Considered preferably, such substances include H _TwoO_TwoSuch
Active oxygen generator, L-buthionine sulfoximine
(LBSO) and the like. Furthermore, Daxx
(Fasdeath domain-associated protein) and ASK1
It may be a substance that causes the interaction of This
As described above, the pharmaceutical composition for treating cancer according to the invention of
s_TwoO_ThreeAnd other ASK1-SEK1-JNK cascades
The drug is used in combination with
Can be selected according to the method of administration to the patient. example
If the effective amount of As_TwoO_ThreeAnd ASK1-SEK1-J
Pharmaceutically acceptable carrier for NK cascade activator
And a mixture produced by homogenization. This
At this point, the carrier should be compatible with the form of preparation desired for administration.
Thus, it can take various forms. Preferably,
These preparations can be taken orally or by injection or infusion
Those in a unit dosage form that can be administered more are exemplified.
Of course, depending on the form and location of the cancer,
The preparation may take the form as given. This out
When preparing the cancer therapeutic preparation of the present invention for oral administration
In addition to liquid preparations such as suspensions and syrups,
Powders, pills, capsules, tablets and the like can be used. Sutra
For oral liquid preparations, the above effective amount of As_TwoO_ThreeAnd AS
In addition to the K1-SEK1-JNK cascade activator
Water, sugars (eg, sucrose, fructose, etc.),
Glycols (eg, polyethylene glycol), oils
(Eg, soybean oil), preservatives (eg, alkyl parahi
Droxybenzoate, etc.), flavors and sweeteners (eg, strike
Roberry flavor, peppermint, etc.)
You may. Powders, pills, capsules, tablets,
Excipients (eg lactose, glucose, mannitol
Etc.), disintegrants (eg starch, sodium alginate)
), Lubricants (eg, talc, etc.), binders (eg,
Roxypropylcellulose, gelatin, etc.), surfactant
(Such as fatty acid esters), plasticizers (such as glycerol)
The effective amount of As_TwoO_ThreeAnd ASK1-SE
In addition to the K1-JNK cascade activator, conventional methods
It is manufactured by Furthermore, as a formulation solution for injection
Is a salt solution, a glucose solution, or a mixture thereof
The above effective amount of As_TwoO_ThreeAnd ASK1-SE
It can be prepared by adding a K1-JNK cascade activator.
Can be. The preparation for treating cancer of the present invention is orally administered.
Preferably administered or injected,
The effective dose is 10-100 mg / person / day,
The frequency of administration is preferably 1 to 3 times a day. Less than
Examples will be shown below, and the embodiments of the present invention will be further described.
Will be described in detail. Of course, the present invention is limited to the following examples.
It is not specified, and various aspects can be
It goes without saying that it is capable.

EXAMPLES In Reference Examples 1 and 2 below, cell staining was performed.
The color was determined by the following operation. First, 8
% Formaldehyde (final formaldehyde concentration
4%), left for 20 minutes, and then
The cells were obtained by centrifugation at 00 rpm for 10 minutes. this
Was washed with PBS, and 1% bovine serum albumin (blocky
Blocking with PBS
After that, each primary antibody (mouse monoclonal anti-PML anti-
Body (manufactured by Santa Cruz), rabbit polyclonal anti-Daxx
Antibody (Sumitomo Electric), rabbit polyclonal anti-ASK1
Antibodies (Saitoh, M. et al., EMBO J. 17, 2596-2606 (199
8)), or rabbit polyclonal anti-phosphorylation site specific
1: 1000 diluted for all SEK1 antibodies (NEB)
) At 4 ° C overnight. Got
Cells can be further conjugated to FITC and / or Texas Red
With secondary antibody (1: 1000 dilution from Vector Laboratory)
Both were incubated at room temperature for 4 hours, and DAPI (Ve
ctor Laboratory). <Reference Example 1>PMLbo in APL cells (NB4)
Accumulation of activated SEK1 in dy 1 μM As_TwoO_ThreeNB4 after 12 hours
Bubbles and As_TwoO_ThreeThe cells before addition are treated with anti-PML antibody and anti-phosphorus.
Oxidation site-specific SEK1 antibodies
Staining method (Dyck, J.A. et al., Cell 76, 333-343 (199
Co-immunostaining was performed according to 4)) and compared. Figure of fluorescence microscope image
1 is shown. As_TwoO_Three12 hours after the addition of
That activated SEK1 is present in the PMLbody
It has been certified. (FIGS. 1 (a) and (b)) In addition, simultaneous staining with DAPI allowed
These DNAs are aggregated and fragmented in
Cells suggest apoptosis-like cell death
Was. (FIG. 1 (c)) <Reference Example 2>Activation of SEK1 by As ₂ O ₃  1 μM As_TwoO_ThreeNB4 after 12 hours
Bubbles and As_TwoO_ThreeView cells before addition with a confocal laser microscope.
I thought. The microscope images are shown in FIGS. (1) Localization of Daxx: As_TwoO_ThreeWithout adding NB4
In cells, Daxx, in a wide range of NB4 cells, PM
It was confirmed that they coexist with L in the form of micro spots. (Figure 1
(A-1), (b-1)) On the other hand, As_TwoO_ThreeNB4 cells reconstituted in the nucleus
Daxx was localized in the resulting PMLbody. (FIG. 1 (a
-2), (b-2)) Also, as previously reported (Zhong, S. et.
 al., J. Exp..Med. 191, 631-640 (2000)), Retinoi
Reconstitution in NB4 cells to which acid (RA) was added
It was confirmed that Daxx was localized in the PMLbody
Was. (FIGS. 1 (a-3) and (b-3)) (2) Localization of ASK1 (FIG. 3): Using anti-ASK1 antibody
Observation of the localization pattern of ASK1
_TwoO_ThreeIn NB4 cells without addition of ASK1, ASK1 was mainly
Present in the cytoplasm. (FIGS. 3 (a-1) and (b-1)) On the other hand, As_TwoO_ThreeIn NB4 cells to which ASK1 was added,
Translocation occurs and PMLbody is
Restructuring has taken place. (Fig. 3 (a-2), (b-2)) However, such translocation and ASK1
Of PMLbody by NB4 cells with RA
Was not confirmed. (FIG. 3 (a-3), (b-
3)) <Reference example 3>NB4 cell death by As ₂ O ₃
Importance of SEK1 activation Whether SEK1 Activation is Essential for NB4 Cell Death
To confirm the presence of SEK1, dominant inactivity (dominant
Negative) expressing the mutant (DNSEK1)
The effect of suppressing death was confirmed. First, NB4 cells (Lanotte,
M. et al., Blood 77, 1080-1086 (1997))
RPM with inert FBS (Life Technologies)
The cells were cultured in I medium (manufactured by Life Technologies). This and
The DNA introduction and expression are performed by Gene Pulser Electric
This was performed using a perforator (manufactured by Bio-Rad) (Kakizuka, A.
et al., Cell 66, 663-674 (1991)). 2 × 10⁷Individual details
Vesicles with 10 μg of pEBG-SEK1K-R²⁶(Overt inactive
SEK1), 1 μg of pEGFP-C1 vector
(Manufactured by CLONTECH) and the expression of DNSEK1
Vesicles were obtained. . Four hours after expression, 1 μM As_TwoO_ThreeCultivate
Add to the ground, As_TwoO_ThreeWas added to the medium. Another 6 hours
After the passage, the nuclei were stained with Hoechst 33342, and
Observed with a light microscope. The apoptotic cell abundance is
GFP-SEK1 having an apoptotic aggregation nucleus or
Divide the number of GFP-DNSEK1 by the total number of GFP-cells
And was calculated by: (About 300 pieces)_TwoO_ThreeNB4 in the presence and absence and
Presence of apoptotic cells in DNSEK1-expressing cells
The percentage was shown. As_TwoO_ThreeIn NB4 added with
About 70% of SEK1 cells have apoptotic nuclei
However, in DNSEK1, As_TwoO_ThreeCell death by
Was greatly suppressed. (FIG. 4 (a)) Similarly, it is known that JNK activity is inhibited at about 30 μM.
SB202190 (Jacinto, E., Werlen, G., and Kari
n, M., Immunity 8, 31-41 (1988)) was added to the cells.
In that case, As_TwoO_ThreeCell death was significantly suppressed.
(FIG. 4 (b)) Such inhibition by SB202190 inhibits p38 kinase.
At a concentration of 1 μM,
From, As_TwoO_ThreeCauses NB4 cell death
Requires the SEK1-JNK cell death kinase cascade
It was shown to be. From the above, PMLbody is As_Two
O_ThreeIs involved in cancer cell death by As _TwoO_ThreeTo
Thus, the PMLbody was reconstructed, and the ASK1 PMLbody
And SEK1 is activated on PMLbody
It became clear that cell death was occurring. this
The concentration of As is as low as 0.5-1 μM._TwoO_ThreeBut in
 APL cells die in vitro and in vivo (in vitr
o: Chen, G.Q. et al., Blood 88, 1052-1061 (1996);
Zhu, J. et al., Proc. Natl. Acad. Sci. U.S.A. 94, 39
78-3983 (1997); in vivo: Chen, G.Q. et al., Blood
89, 3345-3353 (1997); Shen, Z.X. et al., Blood 89,
3354-3360 (1997); Soignet, S.L. et al., N. Engl.
J. Med. 339, 1341-1348 (1998)), ASK1-S
This shows that EK1-JNK is activated.
I was instigated. <Example 1>As ₂ O ₃ + LBSO of human cervical cancer cells
Cell death by addition 5 × 10^ThreeHuman cervical (HeLa) cells in 10% serum
Suspended in 100 μl of D-MEM medium containing 96 wells.
Seed on plate, and after 24 hours, DM with 10% serum
Each concentration of As prepared in 100 μl of EM medium_TwoO_ThreeOr
As_TwoO_ThreeAnd LBSO were added to the well. One condition
3 wells were set, and the average value was determined. 2 from drug addition
After 4 hours, WST-8 (Cell Counting Kit, Nakar
ai, Kyoto) and measure the relative cell number.
I got Dose obtained with 100% of cells containing only medium
The response curve is shown in FIG. According to FIG. 5, LBSO is added.
In this way, the sub-cell of HeLa cells depends on the LBSO concentration.
It has been confirmed that susceptibility to cell death by arsenic acid is increased.
Was. <Example 2>As ₂ O ₃ + LBS in prostate and breast cancer cells
Cell death due to O addition The same operation as in Example 1 was carried out by
Gen-reactive human prostate cancer cell line (LNCaP: Horoszew)
icz J.X., Leong, S.S., Kawinski, E., Karr, J.P., Ro
senthal, H., Chu, T.M., Mirand, E.A., Murphy, G.
P., Cancer Res. 43 (4), 1809-18 (1983)), bone metastasis
Androgen refractory human prostate cancer cell line (PC-3:
Perkel, V.S., Mohan, S., Herring, S.J., Baylink,
D.J., Linkhart, T.A., Cancer Res. 50 (21), 6902-9 (1
990)), androgen-refractory human prostate from brain metastasis
Cancer cell line (DU145: Stone, K.R., Mickey, D.D., W
underli, H., Mickey, G.H., Paulson, D.F., Int.
Cancer 21 (3), 274-8, (1978)) and antiandrogen
Responsive mouse breast cancer cells (SC115: Kitamura, Y., Okam
oto, S., Hayata, I., Uchida, N., Yamaguchi, K., Ma
tsumoto, K., CancerRes. 39 (11), 4713-9 (1979))
It was carried out using. 100% of cells containing only medium
From the dose-response curve₅₀(50% cytotoxicity concentration)
Was obtained and shown in FIG. From FIG. 6, 10 μM or 40 μM
The addition of μM LBSO allows the arsenite to induce cancer cell I
C₅₀Can be reduced from several μM to 1 μM or less.
Was. <Example 3>Apot by combined use of As ₂ O ₃ and LBSO
Morphological observation of cis induction 7.5 × 10^ThreeHeLa cells in 10% serum-containing DME
M medium 2 ml, and seeded on a 6-well plate.
After 4 hours, prepare with 2 ml of D-MEM containing 10% serum
0.5 μM of LBSO was added to the well. LB
24 hours after SO addition, 10 minutes with 4% formalin
Fixation and embedding medium containing DAPI (VECTASHIEEL)
D, at Vector Laboratories, Inc. Burlingame, CA)
FIG. 7 shows a 2 μM As sample._TwoO_ThreeOnly and 0.5 μM As
_TwoO_ThreeAnd optical microscope when 0.5 μM LBSO was added
The image and the fluorescence microscope image are shown. From FIG. 7, 2 μM A
s_TwoO_Three0.5 μM As compared to single administration of_TwoO_ThreeAnd 0.
The combined administration of 5 μM LBSO resulted in HeLa cell death
Was found to be effective.

As described above in detail, according to the present invention, low concentrations of As ₂ O ₃ and ASK1 exhibiting no side effects.
-By using the activator of SEK1-JNK cascade in combination, a pharmaceutical for solid cancer treatment capable of killing cancer cells with less burden on patients is provided.

[Brief description of the drawings]

FIG. 1 is a fluorescence microscopic image showing accumulation of activated SEK1 in cancer cells (NB4) before and after addition of As ₂ O ₃ in Reference Example 1 of the present invention. (A) staining with an anti-PML antibody, (b) staining with an anti-phosphorylation site-specific SEK1 antibody,
(C) Staining nuclei with DAPI.

FIG. 2 is a diagram illustrating the localization of Daxx in cancer cells (NB4) before and after addition of As ₂ O ₃ in a reference example of the present invention. (A) Staining with anti-PML antibody (1) As ₂
Before addition of O ₃ , (2) 12 hours after addition of As ₂ O ₃ ,
(3) 48 hours after the addition of retinoic acid (RA);
(B) Staining with rabbit polyclonal anti-Daxx antibody
(1) Before As ₂ O ₃ addition, (2) 12 hours after addition of As ₂ O ₃ , (3) 48 hours after addition of retinoic acid (RA); (c) (1) PML antibody staining image Images obtained by superimposing the stained images with the Daxx antibody, (2) and (3) nuclei were stained with DAPI.

FIG. 3 is a diagram illustrating the localization of ASK1 in cancer cells (NB4) before and after addition of As ₂ O ₃ in Reference Examples of the present invention. (1) Before adding As ₂ O ₃ , (2) As ₂ O ₃
12 hours after the addition, (3) 48 hours after the addition of retinoic acid (RA) (a) Staining with anti-PML antibody, (b)
Stained with rabbit polyclonal anti-ASK1 antibody, (c) D
Stain nuclei with API.

FIG. 4 shows (a) 1 μM of A in a reference example of the present invention.
s_TwoO_ThreeCells (NB4) and SE before and after the addition of
In K1 overt inactive (dominant negative) mutant
Shows the apoptotic cell abundance rate (average value of three times)
Graph and (b) As _TwoO_ThreeSB202190 on cell death by
Effect (30 μM) on the presence of apoptotic cells
It is the graph which showed the rate (average value of three times).

FIG. 5 shows the survival rate of HeLa cells when As ₂ O ₃ alone or As ₂ O ₃ and LBSO were added, and the number of HeLa cells in DMEM medium alone was 10 in Examples of the present invention.
It is a dose-response curve obtained as 0%.

FIG. 6 shows an embodiment of the present invention._TwoO_ThreeOnly
Or As_TwoO_ThreeLNCaP and P when LBSO was added
IC of C3, DU145, and SC115 cancer cells
₅₀(50% cytotoxicity) with cells in DMEM medium only
Is a graph obtained by assuming that the number is 100%.

FIG. 7 shows an embodiment of the present invention in which 2.0 μMAs ₂
O ₃ only or 0.5 μM As ₂ O ₃ +0.5 μM L
(A) Optical microscope image showing cell death of HeLa cells when BSO was added, and (b) a fluorescence microscope image showing nuclei stained with DAPI. (The cells that look round are dead cells.)

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Hidenori Ichijo 3-14-8 Koishikawa, Bunkyo-ku, Tokyo 302 Residence Koishikawa F-term (reference) 4C084 AA19 MA02 NA14 ZB261 ZB262 4C086 AA01 AA02 HA07 HA21 MA02 NA06 NA14 ZB26 4C206 AA01 AA02 JA76 MA02 NA14 ZB26

Claims (8)

[Claims] 1. An arsenous acid and ASK1-SEK other than arsenous acid
A preparation for treating solid cancer, which comprises a 1-JNK cascade activator. 2. The preparation for treating solid cancer according to claim 1, wherein the concentration of arsenous acid is 0.1 to 2 μM. 3. ASK1-SEK1-JN other than arsenous acid
3. The preparation for treating solid cancer according to claim 1, wherein the K cascade activator is a JNK activator. 4. ASK1-SEK1-JN other than arsenous acid
3. The preparation for treating solid cancer according to claim 1, wherein the K cascade activator is an SEK1 activator. 5. ASK1-SEK1-JN other than arsenous acid
3. The preparation for treating solid cancer according to claim 1, wherein the K cascade activator is an ASK1 activator. 6. The preparation for treating solid cancer according to claim 5, wherein the ASK1 activator is a substance that causes the dissociation of thioredoxin from ASK1. 7. The preparation for treating solid cancer according to claim 6, wherein the substance that causes the dissociation of thioredoxin from ASK1 is an active oxygen generating substance. 8. The preparation for treating a solid cancer according to claim 5, wherein the ASK1 activator is L-buthionine sulfoximine.