JPH0436240A - Treating agent for adult t-cell leukemia - Google Patents
Treating agent for adult t-cell leukemiaInfo
- Publication number
- JPH0436240A JPH0436240A JP14034790A JP14034790A JPH0436240A JP H0436240 A JPH0436240 A JP H0436240A JP 14034790 A JP14034790 A JP 14034790A JP 14034790 A JP14034790 A JP 14034790A JP H0436240 A JPH0436240 A JP H0436240A
- Authority
- JP
- Japan
- Prior art keywords
- adult
- digitoxin
- cell leukemia
- treating agent
- drug
- 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
Links
- 208000009746 Adult T-Cell Leukemia-Lymphoma Diseases 0.000 title claims abstract description 8
- 208000016683 Adult T-cell leukemia/lymphoma Diseases 0.000 title claims abstract description 8
- 201000006966 adult T-cell leukemia Diseases 0.000 title claims abstract description 8
- 239000003814 drug Substances 0.000 claims abstract description 18
- WDJUZGPOPHTGOT-OAXVISGBSA-N Digitoxin Natural products O([C@H]1[C@@H](C)O[C@@H](O[C@@H]2C[C@@H]3[C@@](C)([C@@H]4[C@H]([C@]5(O)[C@@](C)([C@H](C6=CC(=O)OC6)CC5)CC4)CC3)CC2)C[C@H]1O)[C@H]1O[C@@H](C)[C@H](O[C@H]2O[C@@H](C)[C@@H](O)[C@@H](O)C2)[C@@H](O)C1 WDJUZGPOPHTGOT-OAXVISGBSA-N 0.000 claims abstract description 17
- 229960000648 digitoxin Drugs 0.000 claims abstract description 17
- WDJUZGPOPHTGOT-XUDUSOBPSA-N digitoxin Chemical compound C1[C@H](O)[C@H](O)[C@@H](C)O[C@H]1O[C@@H]1[C@@H](C)O[C@@H](O[C@@H]2[C@H](O[C@@H](O[C@@H]3C[C@@H]4[C@]([C@@H]5[C@H]([C@]6(CC[C@@H]([C@@]6(C)CC5)C=5COC(=O)C=5)O)CC4)(C)CC3)C[C@@H]2O)C)C[C@@H]1O WDJUZGPOPHTGOT-XUDUSOBPSA-N 0.000 claims abstract description 17
- 229940124597 therapeutic agent Drugs 0.000 claims description 4
- 239000004480 active ingredient Substances 0.000 claims description 3
- 229940079593 drug Drugs 0.000 abstract description 14
- 239000000843 powder Substances 0.000 abstract description 5
- 239000003795 chemical substances by application Substances 0.000 abstract description 4
- 150000001875 compounds Chemical class 0.000 abstract description 2
- 229930182470 glycoside Natural products 0.000 abstract description 2
- 150000002338 glycosides Chemical class 0.000 abstract description 2
- 230000003177 cardiotonic effect Effects 0.000 abstract 1
- 210000000813 small intestine Anatomy 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 18
- LPMXVESGRSUGHW-HBYQJFLCSA-N ouabain Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@@H]1C[C@@]2(O)CC[C@H]3[C@@]4(O)CC[C@H](C=5COC(=O)C=5)[C@@]4(C)C[C@@H](O)[C@@H]3[C@@]2(CO)[C@H](O)C1 LPMXVESGRSUGHW-HBYQJFLCSA-N 0.000 description 8
- 210000000265 leukocyte Anatomy 0.000 description 7
- LPMXVESGRSUGHW-UHFFFAOYSA-N Acolongiflorosid K Natural products OC1C(O)C(O)C(C)OC1OC1CC2(O)CCC3C4(O)CCC(C=5COC(=O)C=5)C4(C)CC(O)C3C2(CO)C(O)C1 LPMXVESGRSUGHW-UHFFFAOYSA-N 0.000 description 6
- LPMXVESGRSUGHW-GHYGWZAOSA-N Ouabain Natural products O([C@@H]1[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O1)[C@H]1C[C@@H](O)[C@@]2(CO)[C@@](O)(C1)CC[C@H]1[C@]3(O)[C@@](C)([C@H](C4=CC(=O)OC4)CC3)C[C@@H](O)[C@H]21 LPMXVESGRSUGHW-GHYGWZAOSA-N 0.000 description 6
- 244000166550 Strophanthus gratus Species 0.000 description 6
- 230000001472 cytotoxic effect Effects 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 229960003343 ouabain Drugs 0.000 description 6
- 239000000523 sample Substances 0.000 description 5
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 4
- 239000008101 lactose Substances 0.000 description 4
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 4
- 230000001988 toxicity Effects 0.000 description 4
- 231100000419 toxicity Toxicity 0.000 description 4
- 231100000002 MTT assay Toxicity 0.000 description 3
- 238000000134 MTT assay Methods 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 229940097217 cardiac glycoside Drugs 0.000 description 3
- 239000002368 cardiac glycoside Substances 0.000 description 3
- 239000000496 cardiotonic agent Substances 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 229930002534 steroid glycoside Natural products 0.000 description 3
- FDWRIIDFYSUTDP-UHFFFAOYSA-N 102850-49-7 Natural products CC1OC(O)CC(O)C1O FDWRIIDFYSUTDP-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 229920002261 Corn starch Polymers 0.000 description 2
- JWFRNGYBHLBCMB-UHFFFAOYSA-N D-Canaytose Natural products CC(O)C(O)C(O)CC=O JWFRNGYBHLBCMB-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 208000033855 HTLV-1 carrier Diseases 0.000 description 2
- 229920002153 Hydroxypropyl cellulose Polymers 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 239000008120 corn starch Substances 0.000 description 2
- 210000004748 cultured cell Anatomy 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- JWFRNGYBHLBCMB-NGJCXOISSA-N digitoxose Chemical compound C[C@@H](O)[C@@H](O)[C@@H](O)CC=O JWFRNGYBHLBCMB-NGJCXOISSA-N 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000001863 hydroxypropyl cellulose Substances 0.000 description 2
- 235000010977 hydroxypropyl cellulose Nutrition 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 235000019359 magnesium stearate Nutrition 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 231100000816 toxic dose Toxicity 0.000 description 2
- AZKSAVLVSZKNRD-UHFFFAOYSA-M 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Chemical compound [Br-].S1C(C)=C(C)N=C1[N+]1=NC(C=2C=CC=CC=2)=NN1C1=CC=CC=C1 AZKSAVLVSZKNRD-UHFFFAOYSA-M 0.000 description 1
- 108091006112 ATPases Proteins 0.000 description 1
- 102000057290 Adenosine Triphosphatases Human genes 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 241000598436 Human T-cell lymphotropic virus Species 0.000 description 1
- 102000043276 Oncogene Human genes 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 208000001388 Opportunistic Infections Diseases 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 102000005393 Sodium-Potassium-Exchanging ATPase Human genes 0.000 description 1
- 108010006431 Sodium-Potassium-Exchanging ATPase Proteins 0.000 description 1
- 208000000389 T-cell leukemia Diseases 0.000 description 1
- 208000028530 T-cell lymphoblastic leukemia/lymphoma Diseases 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 230000007059 acute toxicity Effects 0.000 description 1
- 231100000403 acute toxicity Toxicity 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000036765 blood level Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229940084030 carboxymethylcellulose calcium Drugs 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000001668 nucleic acid synthesis Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229940126701 oral medication Drugs 0.000 description 1
- 239000013610 patient sample Substances 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 229920001592 potato starch Polymers 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 150000008143 steroidal glycosides Chemical class 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
【発明の詳細な説明】
東！上９上月±１
本発明はジキトキシンを有効成分として含有することを
特徴とする成人Ｔ細胞白血病治療剤に関する。[Detailed description of the invention] East! The present invention relates to a therapeutic agent for adult T-cell leukemia characterized by containing diquitoxin as an active ingredient.
従ｊトク乏迷
ジギトキシンは、下記の構造式：
％式％）
（Ｄ−ジギト寿ソース）（Ｄ−ジイトキソース＞（Ｄ−
ジギト臀ソ＝ス）を有する強心配糖体の一種で、経口剤
として用いられてきた１強心配糖体の薬理作用は種類に
依らず極めてよく似ており、心臓に対する作用が主なも
のであるが、やや大量では腎臓や中枢神経系にも作用す
る０作用点は、細胞膜のナトリウムポンプを抑制（Ｎａ
”、Ｋｏ、ＡＴＰａｓｅの抑制）する結果、細胞内Ｃａ
”＋イオンの増大がおこるためと考えられている。この
作用はｍｐ毒性とは区別できていなかった２強心配糖体
の１種である０ｕａｂａｉｎ の細胞毒性に関する報告
は多い。Digitoxin has the following structural formula: % formula %) (D-digitoxose) (D-digitoxose
Cardiac glycoside is a type of cardiac glycoside that has been used as an oral drug.The pharmacological actions of cardiac glycosides are very similar regardless of the type, and the main action is on the heart. However, at moderately high doses, the 0-point of action also acts on the kidneys and central nervous system, inhibiting the sodium pump in the cell membrane (Na
”, Ko, ATPase inhibition), intracellular Ca
This effect is thought to be due to an increase in + ions.This effect could not be distinguished from MP toxicity.There are many reports regarding the cytotoxicity of Ouabain, a type of dicardial glycoside.
０ｕａｂａｉｎはｐｈｙｔｏｈａａｍａｇｇｌｕｔｉｎ
ｉｎ （ＰＨＡ ）でのヒトリンパ球の活性化を抑制し
、この抑制は０ｕａｂａｉｎ添加後直ちにタンパク合成
阻害が、遅れて核酸合成阻害が起きる結果と考えられた
。Ouabain is phytohaamaggglutin
In (PHA), activation of human lymphocytes was suppressed, and this suppression was thought to be the result of immediate protein synthesis inhibition and delayed nucleic acid synthesis inhibition after addition of Ouabain.
０ｕａｂａｉｎの抗！瘍活性については次のような報告
がある１種々のオンコジーン（Ｈ−ｒａｓ、 Ｋ−ｒａ
ｓ。0uabain's resistance! Regarding tumor activity, there are reports on various oncogenes (H-ras, K-ra
s.
ｍｅｔ、 ｒａｆ、 ｆｍｓ、 ｍｏｓ、 ｓｉｓ ）
でトランスホームしたマウスＮＩＨ／３Ｔ３細胞は０ｕ
ａｂａｉｎに対する感受性が高くなったと言う、また、
ヒト細胞（ＨＯ５ｃｅｌｌ １ｉｎｅ）でもに−ｒａｓ
でトランスホームすると同時に０ｕａｂａｉｎに感受性
が高まった。このように０ｕａｂａｉｎは増殖の盛んな
細胞に、より強い毒性を示すことが分かる。しかし、ジ
ギトキシンについてはこのような報告はまだなされてい
ない。met, raf, fms, mos, sis)
Mouse NIH/3T3 cells transformed with 0u
It is said that the sensitivity to abain has increased, and
Human cells (HO5cell 1ine) Moni-ras
The sensitivity to Ouabain increased at the same time as the transformation. Thus, it can be seen that Ouabain exhibits stronger toxicity toward cells that are actively proliferating. However, no such report has been made regarding digitoxin.
明が解決しようとする課
成人Ｔ細胞白血病（Ａｄｕｌｔ Ｔ−ｃｅｌｌ ｌｅｕ
ｋｅｍｉａ（ＡＴＬ） ）はヒトＴ細胞白血病ウィルス
（ＨＴＬＶ−１）の感染に起因して起きる疾病である。Adult T-cell leukemia (Adult T-cell leukemia)
kemia (ATL)) is a disease caused by infection with human T-cell leukemia virus (HTLV-1).
その感染経路としては、母から乳児へ、また輸血などの
感染ノンバ球の混入によるものと考えられている。The route of infection is thought to be from mother to infant, or through contamination with infected non-baby cells such as through blood transfusions.
ＨＴＬＶ−１健康保因者のうち、１５００人に１人の割
合で４０〜６０才代で発症する。ＴｍＰの腫瘍化により
免疫機能低下を来し、多くは日和見感染により死に至る
。しかし、まだ的確な治療薬も存在しないのが現状であ
る。Among healthy HTLV-1 carriers, 1 in 1,500 people will develop the disease between the ages of 40 and 60. Tumorization of TmP causes a decline in immune function, and in many cases, death occurs due to opportunistic infections. However, the current situation is that there is still no exact therapeutic drug.
課 を 決しようとする手段
本発明者らは、公知のジギトキシンが成人Ｔ細胞白血病
細胞阻害作用を有することを発見し、本発明を完成する
に至った。上記のとおり抗ＡＴＬ活性を持つ化合物が待
望きれており、本発明は抗ＡＴＬ治療に大きく貢献する
ものである。The present inventors have discovered that the known digitoxin has an inhibitory effect on adult T-cell leukemia cells, and have completed the present invention. As mentioned above, compounds with anti-ATL activity have been long-awaited, and the present invention will greatly contribute to anti-ATL treatment.
本発明はジギトキシンを有効成分として含有することを
特徴とする成人Ｔ細胞白血病治療剤に関し、その製剤例
としては、錠剤あるいは散剤が挙げられる０例えば、錠
剤１錠中、生薬としてジギトキシンを約０．１ｍｇ含有
させ、賦形剤・結合剤として乳糖、デンプン、結晶セル
ロース、ヒドロキシプロピルセルロースなど、崩壊剤と
してカルボキシメチルセルロースカルシウムなど、滑沢
剤としてステアリン酸マグネシウム、タルクなどを用い
て製剤化することができる。また、散剤１ｇ中、ンギト
キシン０．１ｍｇを含有させ、賦形剤として乳糖、デン
プンなどを用いて製剤化することができる。The present invention relates to a therapeutic agent for adult T-cell leukemia characterized by containing digitoxin as an active ingredient, and examples of its formulation include tablets or powders.For example, one tablet contains about 0.0% of digitoxin as a crude drug. It can be formulated using lactose, starch, crystalline cellulose, hydroxypropyl cellulose, etc. as an excipient/binder, carboxymethyl cellulose calcium, etc. as a disintegrant, and magnesium stearate, talc, etc. as a lubricant. . Further, it can be formulated by containing 0.1 mg of ngitoxin per 1 g of powder and using lactose, starch, etc. as excipients.
ジギトキシンの急性毒性（ＬＤｇｏ）： ７．５ｍｇ／
ｋｇ（マウス経口）
投与方法、患者の年齢、性別、症状により異なるが、成
人に対しては、１回０．０１〜１ｍｇを１Ｂ１ないし３
回経口投与すればよい。Acute toxicity of digitoxin (LDgo): 7.5mg/
kg (mouse oral) Although it varies depending on the administration method, age, sex, and symptoms of the patient, for adults, 0.01 to 1 mg at a time is 1B1 to 3.
It can be administered orally.
（以下余白）
１亙忽
下記組成を有する錠剤および散剤を一般的製法に従って
製剤化した。(Left below) Tablets and powders having the following compositions were prepared according to a general manufacturing method.
（１）錠剤
ジギトキンン ０ 、１ ｒｒ、
ｇ乳糖 ９０ ｍｇ
トウモロコシデンプン 適量ヒドロキシプロ
ピルセルロース ２ ｍｇステアリン酸マグネシ
ウム ２ ｍｇ（２）散剤（１万倍敗）
ジギトキシン
乳糖
バレイショデンブン
トウモロコシデンプン
０．１ｍｇ
４０ ｍｇ
適量
ｇ
合計
ｇ
抗ＡＴＬ活性
材料
薬剤：ジギトキシン（シグマ社、分子量７６４．９２）
およびｏｕａｂａｉｎ （シグマ社：分子量５８４．６
４）を先ずエタノールで溶解し、それを１０倍のＰＢＳ
に溶
解した（５００μＭ）。(1) Tablet digitkin 0, 1 rr,
g lactose 90 mg
Corn starch Appropriate amount Hydroxypropyl cellulose 2 mg Magnesium stearate 2 mg (2) Powder (10,000x loss) Digitoxin Lactose Potato starch Corn starch 0.1 mg 40 mg Appropriate amount g Total g Anti-ATL active material Drug: Digitoxin (Sigma, molecular weight 764.92)
and ouabain (Sigma: molecular weight 584.6
4) was first dissolved in ethanol, and then dissolved in 10x PBS.
(500 μM).
株化細胞：ＡＴＬ由来Ｔ細胞株４株（ＭＴ（、ＭＴ−２
、Ｍｌ−４、Ｈｕτ−１０２） ； ＨＩＬＶ−１キャ
ノア由来Ｔｍ胞株１株（τＬ−５ｕ ）細胞培養：培地
Ｆ ＲＰＭＩ−１６４０＋ １０％ウシ胎児血清を用い
、５％炭酸ガス培養、３
７℃で実験
末梢白血球（Ｐｅｒｉｐｈｅｒａｌ ｂｌｏｏｄ ｌｙ
ｍｐｈｏｃｙｔａ（ＰＢＬ） ）の分離：凝固阻止剤（
ヘパリン）加面液からＰＢＬをＦｉｃｏｌｌ−Ｐａｑｕ
ｅ （７アルマシア社）を用いて分離した６株
化細胞と同じ培地を用いた。但し、
Ｐｈｙｔｏｈａｅｍｍａｇｇｌｕｔｉｎｉｎ ｐ （Ｐ
ＨＡ−ｐ ）を１：１０００添加した。正常人１５人お
よびＡＴＬ患者８人の検体を用いた。Established cell lines: 4 ATL-derived T cell lines (MT (, MT-2)
, Ml-4, Huτ-102) ; 1 HILV-1 Canoa-derived Tm cell line (τL-5u) Cell culture: Using medium F RPMI-1640 + 10% fetal bovine serum, 5% carbon dioxide culture, 37°C Experimental peripheral leukocytes (Peripheral blood ly
mphocyta (PBL)): Separation of anticoagulant (
Ficoll-Paqu (Heparin)
The same medium as used for the 6 cell lines isolated using 7A (Almasia) was used. However, Phytohaemmagglutinin p (P
HA-p) was added at a ratio of 1:1000. Specimens from 15 normal subjects and 8 ATL patients were used.
ＭＴＴアッセイ法による細胞毒活性試験：９６穴プレー
トを用いて、まず培地で薬剤を１０００．５００．２５
０．１２５．６２．５．３１．３．１５．６．７．８そ
して無添加対照のＯｎＭをそれぞれ３ウエルずつ用意し
た。これらに等量（１００μＰ）の細胞浮遊液を加えた
。従って、薬剤の最終濃度は上記の半分となる。培養３
日後、ＭＴＴアッセイを行なった。Cytotoxic activity test using MTT assay method: Using a 96-well plate, first add 1000.500.25 ml of the drug to the culture medium.
0.125.62.5.31.3.15.6.7.8 and OnM as a non-additive control were prepared in three wells each. An equal volume (100 μP) of cell suspension was added to these. Therefore, the final concentration of drug will be half of the above. Culture 3
Days later, MTT assay was performed.
ＭＴＴアッセイ法：３日培養後の各ウェルにＭＴＴ液（
３−（４，５−ジメチルチアゾール−２−イル）−２，
５−ジフェニルテトラゾリウム ブロマイド） ５ ｍ
ｇ／ｍｌ Ｐ Ｂ ５（−））を３０μｉずつ加え、き
らに１時間培養する。各ウェルから培養液を１５０μ！
除去し、代わりに１００μ！の溶解液（２ｍｌ濃塩酸入
り５００ｍ１のインプロパツールで１０％トリトンＸ−
１００としたもの）を加え、結晶状のｆｏｒｍａｓａｎ
（Ｍ Ｔ Ｔから生きた細胞中で形成きれた色素）を
激しく攪拌溶解する。MTT assay method: After 3 days of culture, add MTT solution (
3-(4,5-dimethylthiazol-2-yl)-2,
5-diphenyltetrazolium bromide) 5 m
Add 30 μi of PB 5(-)) to the seedlings and culture for 1 hour. 150μ of culture solution from each well!
Removed and replaced with 100μ! solution (10% Triton X-
100) and add crystalline formasan.
(Dye formed in living cells from MTT) is dissolved by stirring vigorously.
各ウェルを５５０ｎｍ波長（対照：６３０ｎｍ）で吸光
度（０ｐｔｉｃａｌ ｄｅｎｓｉｔｙ ： ＯＤ ）を
測定する。The optical density (OD) of each well is measured at a wavelength of 550 nm (control: 630 nm).
薬剤最終濃度５００ｎＭ（最高濃度）のウェルの測定値
をブランク（ｏ、ｏ ｏ ｏ ）と設定した。このウェ
ルは薬剤により細胞が１００％死に至る対照になる。The reading of the well with a final drug concentration of 500 nM (highest concentration) was set as blank (o, o o o ). This well serves as a control where the drug kills 100% of the cells.
５０％細胞毒性濃度（ＣＣ，。）の計算法＝（１）各薬
剤濃度毎の３ウエルの平均ＯＤ値を求める。Calculation method for 50% cytotoxic concentration (CC, .) = (1) Calculate the average OD value of 3 wells for each drug concentration.
（２）薬剤濃度Ｏｎ Ｍ （対照）のＯＤ値（Ａ）を細
胞毒活性０％、薬剤濃度５００ ｎＭのＯＤ値（Ｂ）で
細胞毒活性を１００％とした。その他の薬剤濃度のＯＤ
値を（Ｃ）としたときの細胞毒活性は（Ａ−Ｃ）／（Ａ
−Ｂ）Ｘｉ ｏ ｏ％と計算きれる。（３）全ての薬剤
濃度において（２）の細胞毒活性％が得られたら、その
活性値５０％を挾む活性値をＹｌおよびＹ２とし、これ
に対応する薬剤濃度（ｎ Ｍ ）をＸｌおよびＸ２とす
る。(2) The OD value (A) of drug concentration On M (control) was taken as 0% cytotoxic activity, and the OD value (B) of drug concentration 500 nM was taken as 100% cytotoxic activity. OD of other drug concentrations
When the value is (C), the cytotoxic activity is (A-C)/(A
-B) It can be calculated as Xi o o%. (3) When the cytotoxic activity % of (2) is obtained at all drug concentrations, the activity values that sandwich the 50% activity value are defined as Yl and Y2, and the corresponding drug concentrations (n M ) are defined as Xl and Y2. Let it be X2.
（以下余白）
（４）ｃｃｓ。の計算式は
結果：
ａ）正常人対照ＰＢＬ
対照として１５人の１０ｍ１ヘパリン加血液を採り、そ
のＰＢＬを用いた結果を下記表１に示す。(Left below) (4) ccs. The calculation formula is the result: a) Normal human control PBL 10 ml of heparinized blood was collected from 15 people as a control, and the results using the PBL are shown in Table 1 below.
（以下余白） ム ｂ）ＡＴＬ患者ＰＢＬ ８検体のＡＴＬ患者由来のＰＢＬの結果を表２に示す。(Margin below) Mu b) ATL patient PBL Table 2 shows the results of PBL derived from 8 ATL patients.
（以下余白） Ｃ）組織培養細胞株 ＡＴＬ由来細胞株４種とＨＴＬＶ ア由来細胞株１種の結果を表３に示す。(Margin below) C) Tissue culture cell line Four ATL-derived cell lines and HTLV Table 3 shows the results for one cell line derived from A.
表１
１キヤリ
ンギトキンンの強心剤としての臨床経験からその有効血
中濃度は１３〜３２．７ ｎＭで、中毒量（以下余白）
は３２．７ ｎＭ以上である。このことを基にして結果
を考察すると、正常人対照データでは試料４および１４
の検体以外は、ＣＣ５゜値は中毒量の範囲に入る結果と
なった。即ち、固体毒性（ｉｎｖｉｖｏ ）と細胞毒性
（ｉｎ ｖｉｔｒｏ ）の結果がよく合致した結果とな
った。しかし、ジギトキシンの毒性には個人差が存在す
ることも示きれ、生体投与時にはこの点注意が肝要であ
る。事実、強心剤として使用される時も、血中濃度をモ
ニターして十分な注意を払って行なわれている。Table 1 Based on clinical experience as a cardiotonic agent, its effective blood concentration is 13 to 32.7 nM, and the toxic dose (hereinafter referred to as the margin) is 32.7 nM or more. Considering the results based on this, the normal control data shows that samples 4 and 14
Except for the samples above, the CC5° values were within the toxic dose range. That is, the solid toxicity (in vivo) and cytotoxicity (in vitro) results were in good agreement. However, it has been shown that there are individual differences in the toxicity of digitoxin, and it is important to pay attention to this point when administering it to living organisms. In fact, even when it is used as a cardiotonic agent, blood levels are monitored and sufficient care is taken.
ＡＴＬ患者ＰＢＬのデータについて見ると、試料７の検
体以外はジギトキンンの臨床的有効濃度の範囲内の値と
なった。また、正常人対照の範囲の値であった検体７を
省いた平均ＣＣ，。は２６゜１±３．４ｎＭで、正常人
対照と比較すると有意に低い値となった。正常人対照と
同様に、検体７で見られた個人差が何に由来するかのか
は不明である０表２の臨床データによれば検体７が特別
正常に近い検体とは考えられなかった。Looking at the data for ATL patient PBL, all samples other than sample 7 had values within the clinically effective concentration range of digitokin. Also, the average CC, excluding sample 7, which had a value in the range of normal human controls. was 26°1±3.4 nM, which was significantly lower than that of normal human controls. As with normal human controls, it is unclear what causes the individual differences observed in Sample 7. According to the clinical data in Table 2, Sample 7 was not considered to be a sample that is particularly close to normal.
組織培養細胞株のデータでは、ジギトキシンの強心剤と
しての臨床的有効量の範囲より低い値となった。ＡＴＬ
由来細胞株とＨＴＬＶ−１キャリア由来細胞株の結果に
有意差は認められなかった。これは、株化Ｉ５た組織培
養細胞では由来が正常であってもｉｎ ｖｉｔｒｏで腫
瘍化している為と考えられる。患者検体よりも培養細胞
株のほうが、ジギトキシンに対してより高い感受性を示
した。Data from tissue culture cell lines indicate values below the range of clinically effective doses of digitoxin as a cardiotonic agent. ATL
No significant difference was observed between the results of the derived cell line and the HTLV-1 carrier derived cell line. This is considered to be because the tissue culture cells of cell line I5 turn into tumors in vitro even if they are of normal origin. Cultured cell lines were more sensitive to digitoxin than patient samples.
これは培養細胞株が均一な細胞集団の為と考えられる。This is considered to be because the cultured cell line is a homogeneous cell population.
以上のことから、ジギトキシンは抗ＡＴＬ剤として有用
であると充分考えられる。また、対照薬として用いた０
ｕａｂａｉｎは消化管での吸収が殆ど認められず、経口
投与が不可能であることも治療薬としては適していない
が、ジギトキシンはノＪ＼膓からの吸収が良く、経口投
与も可能である。従ってジギトキシンは抗ＡＴＬ剤とし
ての臨床応用の可能性が示唆される。From the above, digitoxin is fully considered to be useful as an anti-ATL agent. In addition, 0 used as a control drug
Uabain is hardly absorbed in the gastrointestinal tract and cannot be administered orally, making it unsuitable as a therapeutic agent, but digitoxin is well absorbed from the stomach and can be administered orally. Therefore, the possibility of clinical application of digitoxin as an anti-ATL agent is suggested.
Claims (1)
徴とする成人Ｔ細胞白血病治療剤。(1) A therapeutic agent for adult T-cell leukemia characterized by containing digitoxin as an active ingredient.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14034790A JPH0436240A (en) | 1990-05-29 | 1990-05-29 | Treating agent for adult t-cell leukemia |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14034790A JPH0436240A (en) | 1990-05-29 | 1990-05-29 | Treating agent for adult t-cell leukemia |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0436240A true JPH0436240A (en) | 1992-02-06 |
Family
ID=15266717
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP14034790A Pending JPH0436240A (en) | 1990-05-29 | 1990-05-29 | Treating agent for adult t-cell leukemia |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0436240A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006029020A2 (en) | 2004-09-02 | 2006-03-16 | Bionaut Pharmaceuticals, Inc. | Treatments of refractory cancers using na+/k+-atpase inhibitors |
| JP2008122403A (en) * | 2000-12-04 | 2008-05-29 | Cascade Microtech Inc | Method for assembly of wafer probe |
| WO2010032582A1 (en) | 2008-09-22 | 2010-03-25 | 国立大学法人鹿児島大学 | Therapeutic agent for adult t-cell leukemia |
-
1990
- 1990-05-29 JP JP14034790A patent/JPH0436240A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008122403A (en) * | 2000-12-04 | 2008-05-29 | Cascade Microtech Inc | Method for assembly of wafer probe |
| WO2006029020A2 (en) | 2004-09-02 | 2006-03-16 | Bionaut Pharmaceuticals, Inc. | Treatments of refractory cancers using na+/k+-atpase inhibitors |
| WO2006029020A3 (en) * | 2004-09-02 | 2006-06-01 | Bionaut Pharmaceuticals Inc | Treatments of refractory cancers using na+/k+-atpase inhibitors |
| WO2010032582A1 (en) | 2008-09-22 | 2010-03-25 | 国立大学法人鹿児島大学 | Therapeutic agent for adult t-cell leukemia |
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