JP2002069005A - Solid carcinoma-curing formulation - Google Patents
Solid carcinoma-curing formulationInfo
- Publication number
- JP2002069005A JP2002069005A JP2000261674A JP2000261674A JP2002069005A JP 2002069005 A JP2002069005 A JP 2002069005A JP 2000261674 A JP2000261674 A JP 2000261674A JP 2000261674 A JP2000261674 A JP 2000261674A JP 2002069005 A JP2002069005 A JP 2002069005A
- Authority
- JP
- Japan
- Prior art keywords
- ask1
- cancer
- sek1
- cells
- activator
- Prior art date
- Legal status (The legal status 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 status listed.)
- Pending
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Abstract
Description
【発明の属する技術分野】この出願の発明は、癌の治療
用製剤に関するものである。さらに詳しくは、この出願
の発明は、副作用がなく、患者への負担も少なく、効果
的に癌細胞を死滅させる固形癌治療用製剤に関するもの
である。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.
【従来の技術とその課題】癌による死者は、1990年
には世界で約520万人であったが、2010年には、
800万人に近づくと推計されており(Pisani, P., Pa
rkin, D.M., Bray, F., Ferlay, J., Int. J. Cancer 8
3, 18-29 (1999))、日本国内でも1999年に癌死者
の数は29万人を超えた(厚生省 平成11年人口動態
統計)。その部位別の割合は、胃癌、肺癌が高く、次い
で肝臓癌となっている。癌対策は世界的にも疾病対策に
おいて最重要課題の1つと位置づけられており、その治
療薬や治療法についての研究は様々な角度からなされて
いる。癌の治療法としては、これまで、抗癌剤の投与に
より癌細胞を死滅させる化学療法や、患部を切除する外
科療法、患部に放射線を照射する放射線治療、さらに
は、白血球型の一致した他人から正常な造血幹細胞を提
供してもらい、白血病に侵された骨髄を正常な骨髄に置
換する造血幹細胞移植(骨髄移植)などが一般的に行わ
れている。化学療法において使用される、フルオロウラ
シルなどの代謝拮抗剤、シクロホスファミドなどのアル
キル化剤、塩酸ダウノルビシンやブレオマイシンなどの
抗生物質、植物アルカロイドなどの抗癌剤は、いずれも
治療効果と同時に、悪心、嘔吐、発熱、白血球や血小板
の減少、脱毛、口内炎などの強い副作用を伴う。また、
外科療法は、癌が特定部位に限定して存在している早期
段階では効果を示すが、癌が進行し、転移した場合には
効果が少ない。放射線療法も、同様に、局所療法とい
え、全身性の癌には適さない。造血幹細胞移植は、白血
球型の一致した他人からの正常造血幹細胞の提供を必要
とし、その一致率は、兄弟で25%、両親や親族では1
%以下と低い上、全身麻酔等の負担を提供者にも与え
る。このように、従来行われてきた癌治療法はいずれ
も、効果と同程度、あるいはそれ以上に患者への負担が
大きかったのが実情である。近年、t(15;17)ポ
ジティブな急性前骨髄球性白血病(APL)細胞が亜砒
酸(As2O3)によって死滅することが報告され(Che
n, G.Q. et al., Nat. Genet. 20, 259-265 (1998); Zh
u, J. et al., Proc. Natl. Acad. Sci. U.S.A. 94, 39
78-3983 (1997))、新たな癌治療法として注目されてい
る。これらの報告によれば、As2O3は、APL内にお
いて、PML−RARα(レチノイン酸レセプター(R
AR)αとPML(前骨髄球性白血病細胞)の融合タン
パク質)の劣化を生じさせ、PMLの発癌性ドメインで
あるPMLbodyの再構築を引き起こし、その凝集体の蓄
積により、APLの細胞死が起こる。また、このような
APL細胞の細胞死は、1μM以下のAs2O3で十分に
起こることが分かっている。一方、固形癌については、
前記のとおり、癌死の9割以上が固形癌によるものであ
る(厚生省 平成11年人口動態統計)にも関わらず、
As2O3によって癌細胞を死滅させるためには、大量
(数μM以上)の投与が必要となり、毒性等の危険を伴
うため、APLで期待されるような癌治療の効果は望め
ないと考えられていたのが実情である。そこで、この出
願の発明は、以上のとおりの事情に鑑みてなされたもの
であり、従来技術の問題点を解消し、毒性や副作用がな
く、患者への少ない負担で、固形癌を効果的に治療する
ことができる固形癌治療用製剤を提供することを課題と
している。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.
【課題を解決するための手段】発明者らは、これまで
に、少なくとも8種類の神経変性疾患(ハンチントン
病、マシャドジョセフ病など)において共通する病原性
分子と考えられるポリグルタミンが、PMLbodyの核に
蓄積され、SEK1−JNK(SAPK(stress-activa
ted protein kinase)/ERK(extracellular signal-r
egulated protein kinase) Kinase−c-Jun
NH2−terminal Kinase)細胞死キナー
ゼカスケードを活性化することを培養したPC12神経
細胞における試験より確認し、報告している(Yasuda,
S. et al., Genes Cells 4, 743-756(1999))。そし
て、発明者らは、As2O3によるAPLの細胞死におい
て観察される現象等が、このようなPC12神経細胞で
のポリグルタミンによる細胞死のそれとよく似ているこ
とに着目した。また、発明者らは、ポリグルタミンによ
る神経細胞の細胞死において活性化されたSEK1がP
MLbody内に蓄積されるように、As2O3を添加したA
PL細胞、とくにNB4細胞(Lanotte, M. et al., Bl
ood 77, 1080-1076 (1997))においても、PMLbody内
での活性化SEK1の蓄積が見られることを確認した
(参考例1〜3参照)。これらの知見から、APL細胞
においては、As2O3によって生じるPMLbodyの再構
成が細胞死を引き起こしていると考え、PMLbodyの再
構成を生じさせる、あるいは促進するような活性化物質
を癌細胞中に共存させることにより、癌細胞を死滅させ
ることができると考えた。そして、発明者らは、鋭意研
究を重ね、癌細胞におけるPMLbodyの蓄積機構の重要
な一端を解明し、この出願の発明を完成した。この出願
の発明は、以上のとおりの経緯によりなされたものであ
って、従来技術の課題を解決するものとして、第1に
は、亜砒酸と亜砒酸以外のASK1−SEK1−JNK
カスケード活性化物質を含むことを特徴とする固形癌治
療用製剤を提供する。第2には、この出願の発明は、亜
砒酸の濃度が0.1〜2μMである前記の固形癌治療用
製剤を提供する。この出願の発明は、前記のいずれかの
固形癌治療用製剤において、第3には、亜砒酸以外のA
SK1−SEK1−JNKカスケード活性化物質がJN
K活性化物質であること、第4には、亜砒酸以外のAS
K1−SEK1−JNKカスケード活性化物質がSEK
1活性化物質であること、第5には、亜砒酸以外のAS
K1−SEK1−JNKカスケード活性化物質がASK
1活性化物質であることをその好ましい態様として提供
する。さらに、第6には、この出願の発明は、前記第5
の発明において、ASK1活性化物質がASK1からの
チオレドキシンの解離を起こさせる物質であること、第
7には、このようなASK1からのチオレドキシンの解
離を起こさせる物質が活性酸素発生物質であることを特
徴とする固形癌治療用製剤を提供する。そして、この出
願の発明は、第8には、前記第5の発明において、AS
K1活性化物質がL−ブチオニンスルホキシミンである
固形癌治療用製剤をも提供する。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
NHTwo-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 inventorsTwoOThreeOf 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, AsTwoOThreeA 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, AsTwoOThreeOf 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.
【発明の実施の形態】この出願の発明の固形癌治療用製
剤は、発明者らの鋭意研究によって明らかにされた癌細
胞の細胞死の機構に基づき、固形癌を治療するものであ
る。つまり、この出願の発明の固形癌治療用製剤は、A
SK1−SEK1−JNKカスケード活性化物質によ
り、活性化されたASK1−SEK1−JNKカスケー
ドが、癌細胞の死滅を引き起こすという癌細胞の細胞死
の機構を応用するものである。前述のとおり、As2O3
は、白血病細胞に対しては、1μMという低濃度でも細
胞死を誘導することが知られていたが、これまで、固形
癌については、As2O3を高濃度で添加する必要がある
と考えられていた。この出願の発明では、発明者らの鋭
意研究により、癌細胞の細胞死の機構が明らかにされた
ことにより、ASK1−SEK1−JNKカスケード活
性化物質をAs2O3と併用することにより、低濃度のA
s2O3でも効果的に固形癌を死滅させることができる固
形癌の治療用製剤が提供される。したがって、この出願
の発明の固形癌治療用製剤は、少なくとも亜砒酸(As
2O3)と亜砒酸以外のASK1−SEK1−JNKカス
ケード活性化物質を含むものであればよく、その形態、
形状、摂取方法等はとくに限定されない。このような固
形癌治療用製剤において、As2O3の濃度は、0.1〜
2μMであることが好ましい。さらに好ましくは、0.
5〜1μMである。As2O3の添加量が0.1μM未満
では、癌細胞の細胞死が起こらない場合があり、2μM
よりも多い場合には、毒性が強くなり正常な細胞や組織
をも破壊したり患者に苦痛を与えたりするため好ましく
ない。この出願の発明の固形癌治療用製剤において、A
SK1−SEK1−JNKカスケード活性化物質は、ど
のようなものであってもよく、とくに限定されない。例
えば、JNK(c-Jun NH2-terminal kinase)活性化物
質、SEK1(SAPK (stress-activated protein kinas
e) / ERK (extracellular signal-regulated protein k
inase) kinase)活性化物質、あるいはASK1(apopt
osis signal-regulating kinase 1)活性化物質などが
挙げられる。JNK活性化物質としては、SEK1活性
化物質やASK1活性化物質が考えられているが、JN
K活性化阻害因子の働きを抑制するような物質であって
もよい。また、SEK1活性化物質としては、例えば、
過酸化水素(H2O2)などが挙げられるが、SEK1活
性化阻害因子の働きを抑制する種々の物質などであって
もよい。さらに、ASK1活性化物質としては、ASK
1からチオレドキシンの解離を起こさせるような物質が
好ましく考慮され、このような物質として、H 2O2など
の活性酸素発生物質、L−ブチオニンスルホキシミン
(LBSO)などが例示される。さらには、Daxx
(Fas death domain-associated protein)とASK1
の相互作用を起こさせるような物質であってもよい。こ
の出願の発明の癌の治療用製薬は、以上のとおりに、A
s2O3とそれ以外のASK1−SEK1−JNKカスケ
ード活性化物質を併用するものであるが、その製薬形態
は、患者への投与方法に応じて選択可能である。例え
ば、前記の有効量のAs2O3とASK1−SEK1−J
NKカスケード活性化物質を、薬理的に許容される担体
と混合し、均一化して製造されるものが挙げられる。こ
のとき、担体は、投与に対して望ましい製剤の形態に応
じて、様々な形態をとることができる。好ましくは、こ
れらの製剤としては、経口的に、あるいは注射や点滴に
より投与しうる単位服用形態にあるものが例示される。
もちろん、癌の形態や発症部位によっては、経皮的に投
与されるような形態をとる製剤であってもよい。この出
願の発明の癌治療製剤を経口投与用として調製する場合
には、懸濁液やシロップ剤のような液体調製物の他に、
散剤、丸剤、カプセル、錠剤等とすることができる。経
口液体調製物としては、上記の有効量のAs2O3とAS
K1−SEK1−JNKカスケード活性化物質に加え
て、水、糖類(例えばスクロースやフラクトース等)、
グリコール類(例えばポリエチレングリコール等)、油
類(例えば大豆油等)、防腐剤(例えばアルキルパラヒ
ドロキシベンゾエート等)、香料や甘味料(例えばスト
ロベリー・フレーバー、ペパーミント等)などを添加し
てもよい。散剤、丸剤、カプセル、錠剤は、いずれも、
腑形剤(例えばラクトース、グルコース、マンニトール
等)、崩壊剤(例えばデンプン、アルギン酸ソーダ
等)、滑沢剤(例えばタルク等)、結合剤(例えばヒド
ロキシプロピルセルロース、ゼラチン等)、表面活性剤
(例えば脂肪酸エステル等)、可塑剤(例えばグリセリ
ン等)などを上記の有効量のAs2O3とASK1−SE
K1−JNKカスケード活性化物質に加え、通常の方法
により製造される。さらに、注射用の製剤溶液として
は、塩溶液、グルコース溶液、またはこれらの混合液か
らなる担体に上記の有効量のAs2O3とASK1−SE
K1−JNKカスケード活性化物質を加えて調製するこ
とができる。この出願の発明の癌治療用製剤は、経口的
投与または注射投与されるものであることが好ましく、
その有効投与量は、10〜100mg/人/日であり、
投与回数は、1日1〜3回とすることが好ましい。以
下、実施例を示し、この発明の実施の形態についてさら
に詳しく説明する。もちろん、この発明は以下の例に限
定されるものではなく、細部については様々な態様が可
能であることはいうまでもない。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, AsTwoOThree
Has a low concentration of 1 μM against leukemia cells.
It has been known to induce spore death.
For cancer, AsTwoOThreeNeed 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 AsTwoOThreeLow concentration of A
sTwoOThreeBut 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
TwoOThree) 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, AsTwoOThreeThe concentration of
Preferably it is 2 μM. More preferably, it is 0.
5-1 μM. AsTwoOThreeLess 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-JunNHTwo-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 (HTwoOTwo), 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 TwoOTwoSuch
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
sTwoOThreeAnd 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 AsTwoOThreeAnd 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 AsTwoOThreeAnd 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 AsTwoOThreeAnd 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 AsTwoOThreeAnd 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.
【実施例】以下の参考例1および2において、細胞の染
色は、次のとおりの操作により行った。まず、培地に8
%ホルムアルデヒドを加え(最終ホルムアルデヒド濃度
4%)、20分間放置した後、サイトスピンを用いて6
00rpm、10分間で遠心分離して細胞を得た。これ
をPBSで洗浄し、1%ウシ血清アルブミン(ブロッキ
ングバッファー)を加えたPBSにおいてブロッキング
した後、各1次抗体(マウスモノクローナル抗PML抗
体(Santa Cruz製)、ウサギポリクローナル抗Daxx
抗体(住友電工製)、ウサギポリクローナル抗ASK1
抗体(Saitoh, M.et al., EMBO J. 17, 2596-2606 (199
8))、またはウサギポリクローナル抗リン酸化部位特異
的SEK1抗体(NEB製)いずれも1:1000希
釈)とともに4℃で一晩インキュベートした。得られた
細胞を、さらにFITC複合および/またはTexas Red複合
2次抗体(Vector Laboratory製 1:1000希釈)と
ともに、室温で4時間インキュベートし、DAPI(Ve
ctor Laboratory製)を含む耐退色性溶液に添加した。 <参考例1> APL細胞(NB4)におけるPMLbo
dy中の活性化SEK1の蓄積 1μMのAs2O3を添加して12時間経過後のNB4細
胞とAs2O3添加前の細胞を抗PML抗体および抗リン
酸化部位特異的SEK1抗体で、それぞれ免疫細胞学的
染色法(Dyck, J.A. et al., Cell 76, 333-343 (199
4))により共免疫染色し、比較した。蛍光顕微鏡像を図
1に示した。As2O3の添加から12時間で、再構築し
たPMLbodyに活性化したSEK1が存在することが確
認された。(図1(a)、(b)) また、DAPIによる同時染色により、これらの細胞内
で凝集し、断片化したDNAから、形態学的にこれらの
細胞がアポプトーシス様の細胞死に至ることが示唆され
た。(図1(c)) <参考例2> As2O3によるSEK1の活性化 1μMのAs2O3を添加して12時間経過後のNB4細
胞とAs2O3添加前の細胞を共焦点レーザー顕微鏡で観
察した。顕微鏡像を図2、図3に示した。 (1)Daxxの局在化:As2O3を添加しないNB4
細胞では、Daxxは、NB4細胞の広い範囲で、PM
Lとミクロ斑点状に共存することが確認された。(図1
(a−1)、(b−1)) 一方、As2O3を添加したNB4細胞では、核で再構築
したPMLbodyに、Daxxが局在化した。(図1(a
−2)、(b−2)) また、これまでに報告されているように(Zhong, S. et
al., J. Exp..Med. 191, 631-640 (2000))、レチノイ
ン酸(RA)を添加したNB4細胞においても、再構築
したPMLbodyにDaxxが局在化する様子が確認され
た。(図1(a−3)、(b−3)) (2)ASK1の局在化(図3):抗ASK1抗体を用
いてASK1の局在化パターンを観察したところ、As
2O3を添加しないNB4細胞では、ASK1は、主に細
胞質に存在した。(図3(a−1)、(b−1)) 一方、As2O3を添加したNB4細胞では、ASK1の
トランスロケーションが生じ、細胞核中でPMLbodyの
再構築が起こった。(図3(a−2)、(b−2)) しかし、このようなトランスロケーションとASK1に
よるPMLbodyの再構築は、RAを添加したNB4細胞
では確認されなかった。(図3(a−3)、(b−
3)) <参考例3> As2O3によるNB4の細胞死における
SEK1活性化の重要性 NB4の細胞死においてSEK1の活性化が必須か否か
を確認するために、SEK1の顕性不活性(ドミナント
ネガティブ)変異体(DNSEK1)を発現させ、細胞
死抑制効果を確認した。まず、NB4細胞(Lanotte,
M. et al., Blood 77, 1080-1086 (1997))を10%熱
不活性FBS(Life Technologies製)を加えたRPM
I培地(Life Technologies製)で培養した。このと
き、DNA導入・発現は、Gene Pulser電気
穿孔装置(Bio-Rad製)を用いて行った(Kakizuka, A.
et al., Cell 66, 663-674 (1991))。2×107個の細
胞を10μgのpEBG−SEK1K−R26(顕性不活
性SEK1)で、1μgのpEGFP−C1ベクター
(CLONTECH製)とともに発現させ、DNSEK1発現細
胞を得た。。発現から4時間後、1μM As2O3を培
地に添加し、As2O3を培地に添加した。さらに6時間
経過後、核をHoechst33342で染色して、蛍
光顕微鏡で観察した。アポプトチック細胞の存在率は、
アポプトチック凝集核を有するGFP−SEK1または
GFP−DNSEK1の数をGFP−細胞全数で割るこ
とによって算出された。(約300個) 図4にAs2O3存在下および非存在下でのNB4および
DNSEK1発現細胞におけるアポプトチック細胞の存
在率を示した。As2O3を添加したNB4においては、
約70%のSEK1細胞がアポプトチック核を有してい
たが、DNSEK1においては、As2O3による細胞死
が大幅に抑制された。(図4(a)) 同様に、30μM程度でJNK活性を阻害することで知
られるSB202190(Jacinto, E., Werlen, G., and Kari
n, M., Immunity 8, 31-41 (1988))を細胞に添加した
場合には、As2O3による細胞死が大幅に抑制された。
(図4(b)) このようなSB202190による抑制は、p38キナーゼを抑
制するとされている1μMの濃度では見られなかったこ
とから、As2O3によるNB4の細胞死が起こるために
は、SEK1−JNK細胞死キナーゼカスケードが必須
であることが示された。以上より、PMLbodyがAs2
O3による癌細胞の細胞死に関わっており、As 2O3に
よってPMLbodyが再構築され、ASK1のPMLbody
への移行が起こり、PMLbody上でSEK1が活性化さ
れて細胞死が起こっていることが明らかになった。これ
より、0.5〜1μMという低い濃度のAs2O3でもin
vitroおよびin vivoでAPL細胞が死滅する(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))のは、ASK1−S
EK1−JNKが活性化されているためであることが示
唆された。 <実施例1> ヒト子宮頚癌細胞のAs2O3+LBSO
添加による細胞死 5×103のヒト子宮頚(HeLa)細胞を10%血清
入りD−MEM培地100μlに懸濁し、96well
プレートに蒔き、24時間経過後、10%血清入りDM
EM培地100μlで調製した各濃度のAs2O3または
As2O3とLBSOをwellに添加した。1条件につ
き3well設定し、平均値を求めた。薬剤添加から2
4時間経過後にWST−8(Cell Counting Kit, Nakar
ai, Kyoto)を用いて吸光度を測定し、相対的な細胞数
を得た。培地のみの細胞を100%として得られた用量
反応曲線を図5に示した。図5より、LBSOを添加す
ることにより、LBSO濃度依存的にHeLa細胞の亜
砒酸による細胞死の感受性が高められることが確認され
た。 <実施例2> 前立腺癌と乳癌細胞のAs2O3+LBS
O添加による細胞死 実施例1と同様の操作を、リンパ節転移巣由来アンドロ
ゲン反応性ヒト前立腺癌細胞株(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))、骨転移巣由
来アンドロゲン不応性ヒト前立腺癌細胞株(PC−3:
Perkel, V.S., Mohan, S., Herring, S.J., Baylink,
D.J., Linkhart, T.A., Cancer Res.50(21), 6902-9 (1
990))、脳転移巣由来のアンドロゲン不応性ヒト前立腺
癌細胞株(DU145:Stone, K.R., Mickey, D.D., W
underli, H., Mickey, G.H., Paulson, D.F., Int. J.
Cancer 21(3), 274-8, (1978))およびアンドロゲン反
応性マウス乳癌細胞(SC115:Kitamura, Y., Okam
oto, S., Hayata, I., Uchida, N., Yamaguchi, K., Ma
tsumoto, K., CancerRes. 39(11), 4713-9 (1979))を
用いておこなった。培地のみの細胞を100%として得
られた用量反応曲線からIC50(50%細胞障害濃度)
を求め、図6に示した。図6より、10μMまたは40
μMのLBSOの添加により、亜砒酸による癌細胞のI
C50を数μMから1μM以下に低下できることが示され
た。 <実施例3> As2O3とLBSOの併用によるアポト
ーシス誘導の形態的観察 7.5×103のHeLa細胞を10%血清入りDME
M培地2mlに懸濁し、6wellプレートに蒔き、2
4時間経過後、10%血清入りD−MEM2mlで調製
した0.5μMのLBSOをwellに添加した。LB
SO添加から24時間経過後に4%ホルマリンで10分
間固定し、DAPI入り包埋剤(VECTASHIEL
D,Vector Laboratories, Inc. Burlingame, CA)にて
封入し、蛍光顕微鏡にて観察した 図7に2μMのAs2O3のみ、および0.5μMのAs
2O3と0.5μMのLBSOを添加した際の光学顕微鏡
像および蛍光顕微鏡像を示した。図7より、2μMのA
s2O3の単独投与よりも、0.5μM As2O3と0.
5μM LBSOの併合投与の方が、HeLaの細胞死
を効果的に起こすことが確認された。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 AsTwoOThreeNB4 after 12 hours
Bubbles and AsTwoOThreeThe 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. AsTwoOThree12 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 AsTwoOThreeNB4 after 12 hours
Bubbles and AsTwoOThreeView cells before addition with a confocal laser microscope.
I thought. The microscope images are shown in FIGS. (1) Localization of Daxx: AsTwoOThreeWithout 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, AsTwoOThreeNB4 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
TwoOThreeIn NB4 cells without addition of ASK1, ASK1 was mainly
Present in the cytoplasm. (FIGS. 3 (a-1) and (b-1)) On the other hand, AsTwoOThreeIn 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 AsTwoOThreeCultivate
Add to the ground, AsTwoOThreeWas 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)TwoOThreeNB4 in the presence and absence and
Presence of apoptotic cells in DNSEK1-expressing cells
The percentage was shown. AsTwoOThreeIn NB4 added with
About 70% of SEK1 cells have apoptotic nuclei
However, in DNSEK1, AsTwoOThreeCell 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, AsTwoOThreeCell death was significantly suppressed.
(FIG. 4 (b)) Such inhibition by SB202190 inhibits p38 kinase.
At a concentration of 1 μM,
From, AsTwoOThreeCauses NB4 cell death
Requires the SEK1-JNK cell death kinase cascade
It was shown to be. From the above, PMLbody is AsTwo
OThreeIs involved in cancer cell death by As TwoOThreeTo
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.TwoOThreeBut 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 × 10ThreeHuman 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 mediumTwoOThreeOr
AsTwoOThreeAnd 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 × 10ThreeHeLa 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.TwoOThreeOnly and 0.5 μM As
TwoOThreeAnd optical microscope when 0.5 μM LBSO was added
The image and the fluorescence microscope image are shown. From FIG. 7, 2 μM A
sTwoOThree0.5 μM As compared to single administration ofTwoOThreeAnd 0.
The combined administration of 5 μM LBSO resulted in HeLa cell death
Was found to be effective.
【発明の効果】以上詳しく説明したとおり、この発明に
よって、副作用を示さない低濃度のAs2O3とASK1
−SEK1−JNKカスケード活性化物質を併用するこ
とによって、患者の負担が少なく、癌細胞を死滅させる
ことのできる固形癌治療用製薬が提供される。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.
【図1】この発明の参考例1において、As2O3添加前
後の癌細胞(NB4)における、活性化SEK1の蓄積
を示す蛍光顕微鏡像である。(a)抗PML抗体で染
色、(b)抗リン酸化部位特異的SEK1抗体で染色、
(c)DAPIで核を染色。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.
【図2】この発明の参考例において、As2O3添加前後
の癌細胞(NB4)における、Daxxの局在化を例示
した図である。(a)抗PML抗体で染色 (1)As2
O3添加前、(2)As2O3添加後12時間経過、
(3)レチノイン酸(RA)を添加後48時間経過;
(b)ウサギポリクローナル抗Daxx抗体で染色
(1)As2O3添加前、(2)As2O3添加後12時間
経過、(3)レチノイン酸(RA)を添加後48時間経
過;(c)(1)PML抗体の染色像とDaxx抗体で
の染色像を重ねあわせた像、(2)(3)DAPIで核
を染色。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.
【図3】この発明の参考例において、As2O3添加前後
の癌細胞(NB4)における、ASK1の局在化を例示
した図である。(1)As2O3添加前、(2)As2O3
添加後12時間経過、(3)レチノイン酸(RA)を添
加後48時間経過 (a)抗PML抗体で染色、(b)
ウサギポリクローナル抗ASK1抗体で染色、(c)D
APIで核を染色。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.
【図4】この発明の参考例において、(a)1μMのA
s2O3を添加前後における癌細胞(NB4)およびSE
K1顕性不活性(ドミナントネガティブ)変異体におけ
るアポプトチック細胞の存在率(3回の平均値)を示し
たグラフと、(b)As 2O3による細胞死へのSB202190
の添加(30μM)の影響(アポプトチック細胞の存在
率(3回の平均値))を示したグラフである。FIG. 4 shows (a) 1 μM of A in a reference example of the present invention.
sTwoOThreeCells (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 TwoOThreeSB202190 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).
【図5】この発明の実施例において、As2O3のみ、ま
たはAs2O3とLBSOを添加した際のHeLa細胞の
生存率を、DMEM培地のみのHeLa細胞の数を10
0%として得た用量反応曲線である。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%.
【図6】この発明の実施例において、As2O3のみ、ま
たはAs2O3とLBSOを添加した際のLNCaP、P
C3、DU145、およびSC115の各癌細胞のIC
50(50%細胞障害濃度)を、DMEM培地のみの細胞
の数を100%として得たグラフである。FIG. 6 shows an embodiment of the present invention.TwoOThreeOnly
Or AsTwoOThreeLNCaP 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%.
【図7】この発明の実施例において、2.0μMAs2
O3のみ、または0.5μM As2O3+0.5μM L
BSOを添加した際のHeLa細胞の細胞死を示した
(a)光学顕微鏡像と、(b)DAPIで染色した核を
示す蛍光顕微鏡像である。(丸状に見えるものが死滅し
た細胞。)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.)
───────────────────────────────────────────────────── フロントページの続き (72)発明者 一條 秀憲 東京都文京区小石川3−14−8 小石川住 宅302 Fターム(参考) 4C084 AA19 MA02 NA14 ZB261 ZB262 4C086 AA01 AA02 HA07 HA21 MA02 NA06 NA14 ZB26 4C206 AA01 AA02 JA76 MA02 NA14 ZB26 ──────────────────────────────────────────────────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)
1−JNKカスケード活性化物質を含むことを特徴とす
る固形癌治療用製剤。1. An arsenous acid and ASK1-SEK other than arsenous acid
A preparation for treating solid cancer, which comprises a 1-JNK cascade activator.
求項1の固形癌治療用製剤。2. The preparation for treating solid cancer according to claim 1, wherein the concentration of arsenous acid is 0.1 to 2 μM.
Kカスケード活性化物質がJNK活性化物質である請求
項1または2のいずれかの固形癌治療用製剤。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.
Kカスケード活性化物質がSEK1活性化物質である請
求項1または2のいずれかの固形癌治療用製剤。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.
Kカスケード活性化物質がASK1活性化物質である請
求項1または2のいずれかの固形癌治療用製剤。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.
オレドキシンの解離を起こさせる物質である請求項5の
固形癌治療用製剤。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.
起こさせる物質が活性酸素発生物質である請求項6の固
形癌治療用製剤。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.
ルホキシミンである請求項5の固形癌治療用製剤。8. The preparation for treating a solid cancer according to claim 5, wherein the ASK1 activator is L-buthionine sulfoximine.
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2002060460A1 (en) * | 2001-01-31 | 2002-08-08 | Gifu International Institute Of Biotechnology | Remedies for cancer |
WO2004026407A1 (en) * | 2002-09-06 | 2004-04-01 | Institut Curie | Use of inhibitors of compounds involved in the jun kinase (jnk) biochemical cellular pathway for the treatment or diagnosis of liposarcomas |
-
2000
- 2000-08-30 JP JP2000261674A patent/JP2002069005A/en active Pending
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2002060460A1 (en) * | 2001-01-31 | 2002-08-08 | Gifu International Institute Of Biotechnology | Remedies for cancer |
WO2004026407A1 (en) * | 2002-09-06 | 2004-04-01 | Institut Curie | Use of inhibitors of compounds involved in the jun kinase (jnk) biochemical cellular pathway for the treatment or diagnosis of liposarcomas |
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