JPH03157367A - Isoindolinone derivative, cytocidal agent of cervical carcinoma cell comprising same derivative as active ingredient and production same agent - Google Patents
Isoindolinone derivative, cytocidal agent of cervical carcinoma cell comprising same derivative as active ingredient and production same agentInfo
- Publication number
- JPH03157367A JPH03157367A JP1294415A JP29441589A JPH03157367A JP H03157367 A JPH03157367 A JP H03157367A JP 1294415 A JP1294415 A JP 1294415A JP 29441589 A JP29441589 A JP 29441589A JP H03157367 A JPH03157367 A JP H03157367A
- Authority
- JP
- Japan
- Prior art keywords
- organic solvent
- water
- derivative
- agent
- isoindolinone
- 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.)
- Granted
Links
- PXZQEOJJUGGUIB-UHFFFAOYSA-N isoindolin-1-one Chemical class C1=CC=C2C(=O)NCC2=C1 PXZQEOJJUGGUIB-UHFFFAOYSA-N 0.000 title claims description 13
- 238000004519 manufacturing process Methods 0.000 title claims description 6
- 239000004480 active ingredient Substances 0.000 title claims description 3
- 230000000445 cytocidal effect Effects 0.000 title abstract description 5
- 231100000409 cytocidal Toxicity 0.000 title abstract description 3
- 208000019065 cervical carcinoma Diseases 0.000 title abstract 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims abstract description 42
- 239000003960 organic solvent Substances 0.000 claims abstract description 31
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000008346 aqueous phase Substances 0.000 claims abstract description 11
- 238000000605 extraction Methods 0.000 claims abstract description 10
- 239000000126 substance Substances 0.000 claims abstract description 6
- 238000000622 liquid--liquid extraction Methods 0.000 claims abstract description 5
- 238000001704 evaporation Methods 0.000 claims abstract description 4
- 230000022534 cell killing Effects 0.000 claims description 14
- 206010008342 Cervix carcinoma Diseases 0.000 claims description 9
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 claims description 9
- 201000010881 cervical cancer Diseases 0.000 claims description 9
- 239000000284 extract Substances 0.000 claims description 9
- 239000003139 biocide Substances 0.000 claims description 7
- 238000005194 fractionation Methods 0.000 claims description 3
- 238000001953 recrystallisation Methods 0.000 claims description 3
- 238000004587 chromatography analysis Methods 0.000 claims description 2
- 150000001875 compounds Chemical class 0.000 abstract description 7
- 235000001674 Agaricus brunnescens Nutrition 0.000 abstract description 6
- 239000003795 chemical substances by application Substances 0.000 abstract description 4
- 241000123222 Hericium Species 0.000 abstract description 2
- 240000000588 Hericium erinaceus Species 0.000 abstract description 2
- 235000007328 Hericium erinaceus Nutrition 0.000 abstract description 2
- 241000959662 Hydnaceae Species 0.000 abstract description 2
- 235000013399 edible fruits Nutrition 0.000 abstract 1
- 239000000463 material Substances 0.000 abstract 1
- 238000002360 preparation method Methods 0.000 abstract 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 14
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 12
- 239000010410 layer Substances 0.000 description 11
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- 150000002137 ergosterols Chemical class 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000012916 structural analysis Methods 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 241000222501 Agaricaceae Species 0.000 description 1
- 241001327634 Agaricus blazei Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 238000002738 Giemsa staining Methods 0.000 description 1
- 241000222355 Trametes versicolor Species 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000010200 folin Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
- Indole Compounds (AREA)
Abstract
Description
【発明の詳細な説明】
〈産業上の利用分野〉
本発明は、ハリタケ科（Ｈｙｄｎａｃｅａｅ ） 、サ
ンゴハリタケ属（Ｈｅｒｉｃｉｕｍ）のキノコであるヤ
マブシタケ（Ｈｅｒｉｃｉｕｍ ｅｒｉｎａｃｅｕｍ）
の子実体中に存在するイソインドリノン誘導体及び該イ
ンインドリノン誘導体を増動成分とする子宮頚癌細胞の
５９細胞剤並びに該殺細胞剤の製造方法に関する。[Detailed Description of the Invention] <Industrial Application Field> The present invention relates to Hericium erinaceum, which is a mushroom of the family Hydnaceae and the genus Hericium.
The present invention relates to an isoindolinone derivative present in the fruiting bodies of 59, a cell agent for cervical cancer cells containing the inindolinone derivative as an increasing component, and a method for producing the cell killing agent.
〈従来の技術〉
従来、キノコに含まれる化合物及び該化合物の癌細胞に
対する殺細胞効果について複数の報告がある。例えば、
サルノコシカケ科のキノコであるカワラタケ（Ｐｏｌｙ
ｐｏｒｕｓ ｖｅｒｓｉｃｏｌｏｒ）にはエルゴステロ
ール誘導体が含まれており、該エルゴステロール誘導体
には肝臓癌細胞（Ｈｅｐａｔｏｍａ ｃｅｌｌｓ）に対
する殺細胞効果のあることがテトラヘドロン（Ｔｅｔｒ
ａｈｅｄｒｏｎ）３９ 、２７７９〜２７８５ （１９
８３）に報告されている。またハラタケ科のキノコであ
るヒメマツタケ（Ａｇａｒｉｃｕｓ ｂｌａｚｅｉ）に
もエルゴステロール誘導体が含まれており、該エルゴス
テロール誘導体には子宮頚癌細胞に対する殺細胞効果の
あることがフィトケミストリ
（Ｐｈｙｔｏｃｈｅｍｉｓｔｒｙ） ２７ 、２７７７
〜２７８９ （１９８８）に報告されている。そして同
様のことが特公昭４８−６７６６号公報２特開昭５５−
７１７０２号公報及び特開昭５８−６２１１８号公報等
にも報告されている。<Prior Art> There have been several reports regarding compounds contained in mushrooms and the cell-killing effects of these compounds on cancer cells. for example,
Poly
porus versicolor) contains an ergosterol derivative, and this ergosterol derivative has been shown to have a cell-killing effect on liver cancer cells (Hepatoma cells).
ahedron) 39, 2779-2785 (19
83). In addition, Agaricus blazei, a mushroom of the Agaricaceae family, also contains ergosterol derivatives, and these ergosterol derivatives have been shown to have a cytocidal effect on cervical cancer cells (Phytochemistry 27, 2777).
~2789 (1988). The same thing can be said in Japanese Patent Publication No. 48-6766 2.
It has also been reported in JP-A No. 71702 and Japanese Patent Application Laid-Open No. 58-62118.
〈発明が解決しようとする課題〉
しかし、ヤマプシタケについては上記のような報告がな
い、ヤマブシタケに含まれる化合物及びその癌細胞に対
する殺細胞効果については全く報告がないのである。<Problems to be Solved by the Invention> However, there are no reports as mentioned above regarding Yamapushitake, and there are no reports at all regarding the compounds contained in Yamapushitake and their cell-killing effects on cancer cells.
く課題を解決するためのｆ段〉
しかして本発明者らは、以上の如き実情に鑑みヤマブシ
タケに含まれる化合物及びその癌細胞に対する殺細胞効
果について鋭意研究した結果。Step F for Solving the Problems> However, in view of the above-mentioned circumstances, the present inventors have conducted intensive research on compounds contained in Yamabushitake mushrooms and their cell-killing effects on cancer cells.
ヤマブシタケには特定の化学構造から成る新規のイソイ
ンドリノン誘導体が含まれており、該インインドリノン
誘導体は子宮頚癌細胞に対して優れた殺細胞効果を有し
ていて、かかる殺細胞効果を有する剤が所定の抽出処理
でヤマブシタケから効率的に分離され得ることを見出し
た。Yamabushitake contains a novel isoindolinone derivative with a specific chemical structure, and this inindolinone derivative has an excellent cell killing effect on cervical cancer cells. It has been found that the agent having the following properties can be efficiently separated from Yamabushitake by a predetermined extraction process.
すなわち本発明は、下記構造式で示されるイソインドリ
ノン誘導体、及び該インインドリノン誘導体を有効成分
とする子宮頚癌細胞の殺細胞剤、並びにヤマブシタケの
子実体を水及び有機溶媒の均一・系で抽出処理し、その
抽出液から有機溶媒を蒸発して水相を得た後、該水相を
水及び有機溶媒の混合系で液液分配抽出処理して有機溶
媒層を分取し、そして該有機溶媒層から有機溶媒を蒸発
することを骨子とする上記殺細胞剤の製造方法に係わる
。That is, the present invention provides an isoindolinone derivative represented by the following structural formula, a cell-killing agent for cervical cancer cells containing the inindolinone derivative as an active ingredient, and a homogeneous system of water and an organic solvent using the fruiting body of Yamabushitake. After the organic solvent is evaporated from the extract to obtain an aqueous phase, the aqueous phase is subjected to liquid-liquid distribution extraction with a mixed system of water and an organic solvent to separate the organic solvent layer, and The present invention relates to a method for producing the above-mentioned cell killing agent, the gist of which is evaporating the organic solvent from the organic solvent layer.
上記構造式で示されるイソインドリノン誘導体はヤマブ
シタケの子実体を次のように処理することによって得ら
れる。先ず、ヤマブシタケの生成いは乾燥子実体を水及
び壱機溶媒の均−系で抽出処理し、濾過や遠心分離等で
固液分離したその抽出液から有機溶媒を蒸発して水相を
得る。この場合、水及び有機溶媒の均−系としては、８
０〜８５％メタノールやエタノール、８５％アセトン等
がある。抽出は通常室温で行なうが、加熱還流してもよ
く、抽出時間は通常１〜７２時間である。The isoindolinone derivative represented by the above structural formula can be obtained by treating the fruiting body of Yamabushitake mushroom as follows. First, the produced or dried fruiting bodies of Yamabushitake are extracted with a homogeneous system of water and a single solvent, and the organic solvent is evaporated from the extract, which is separated into solid and liquid by filtration, centrifugation, etc., to obtain an aqueous phase. In this case, the homogeneous system of water and organic solvent is 8
Examples include 0-85% methanol, ethanol, and 85% acetone. Extraction is usually carried out at room temperature, but may also be heated under reflux, and the extraction time is usually 1 to 72 hours.
例工ば、８５％エタノール中にヤマブシタケの朱子実体
を加え、ホモジナイズ処理し、これ奢室温で一昼夜放置
した後、濾過して抽出液な得、該抽出液を減圧下に４０
〜４５°Ｃで加熱してエタノールを蒸発することにより
水相を得るのである０次に、該水相を水及び有機溶媒の
混合系で液液分配抽出処理して有機溶媒層を分取し、該
有機溶媒層から有機溶媒を蒸発して乾固物を得る。この
場合、有機溶媒としては、クロロホルム、酢酸エチルジ
エチルエーテル等があるが、収率の点でクロロホルムが
好ましい４例えば、上記水相にクロロホルムを加え、振
盪後、放置して分層したクロロホルム層を分取し、該ク
ロロホルム層を減圧下に４０〜４５°Ｃで加熱してクロ
ロホルムを蒸発することにより乾固物を得るのである。For example, the vermilion fruiting bodies of Yamabushitake were added to 85% ethanol, homogenized, left at room temperature overnight, filtered to obtain an extract, and the extract was heated under reduced pressure for 40 minutes.
An aqueous phase is obtained by heating at ~45°C to evaporate the ethanol.Next, the aqueous phase is subjected to liquid-liquid partition extraction with a mixed system of water and an organic solvent to separate the organic solvent layer. , the organic solvent is evaporated from the organic solvent layer to obtain a dry solid. In this case, organic solvents include chloroform, ethyl acetate diethyl ether, etc., but chloroform is preferable from the viewpoint of yield4.For example, chloroform is added to the above aqueous phase, shaken, and then left to separate into chloroform layers. The chloroform layer is separated and heated at 40 to 45°C under reduced pressure to evaporate the chloroform to obtain a dry product.
上記乾固物はそれ自体が子宮頚癌細胞の殺細胞剤として
有効なものであるが、該乾固物から不純物を除去してそ
の子宮頚癌細胞に対する殺細胞効果を高めるために、該
乾固物をクロマト分画処理するのが好ましく、クロマト
分画処理したものを更に再結晶処理するのがより好まし
い。この場合詳しくは実施例で後述するように、ヘキサ
ン、クロロホルム、クロロホルム／アセトン等を展開溶
媒とするシリカゲルクロマトグラフィー或いは１層クロ
マトグラフィーを用いてクロマト分画処理することがで
き、またクロロホルム／ジエチルエーテルを用いて再結
晶処理することができる。The dried product itself is effective as a cytocidal agent for cervical cancer cells, but in order to remove impurities from the dried product and enhance its cytocidal effect on cervical cancer cells, It is preferable to subject the solid substance to chromatographic fractionation, and it is more preferable to further recrystallize the chromatographed substance. In this case, as will be described in detail later in Examples, chromatographic fractionation can be carried out using silica gel chromatography or single-layer chromatography using hexane, chloroform, chloroform/acetone, etc. as a developing solvent, or chloroform/diethyl ether. can be used for recrystallization treatment.
かくして再結晶処理することにより単離される化合物の
物理化学的性質及び構造解析結果は下記の通りである。The physicochemical properties and structural analysis results of the compound isolated by the recrystallization treatment are as follows.
（１）分子量：４３３
（２）赤外線吸収スペクトル： ３３００−２１３００
．１６８０．１８８０ｃｍ−１
（３）核磁気共鳴スペクトル（’Ｈ−ＮＭＲ）：１．８
１（ｓ）。(1) Molecular weight: 433 (2) Infrared absorption spectrum: 3300-21300
．． 1680.1880cm-1 (3) Nuclear magnetic resonance spectrum ('H-NMR): 1.8
1(s).
１．８８（ｓ）、 ２．１８（ｓ）、 ２．９７（ｔ、
７．３３）。1.88(s), 2.18(s), 2.97(t,
7.33).
３．１４（ｇ）、 ３．５６（ｄ、 ８．７４）、 ３
．８４（ｔ、 ７．３３）。3.14 (g), 3.56 (d, 8.74), 3
．． 84 (t, 7.33).
３．８４（ｓ）、 ４．２０（ｓ）、 ５．３０（ｔ、
８．７４）。3.84 (s), 4.20 (s), 5.30 (t,
8.74).
８．０８（ｓ）、 ８．９８（ｓ）、 ７．２０〜７．
２８（ａ）（４）溶媒に対する溶解性：酢酸エチル、ク
ロロホルム、アセトンに可溶、ヘキサン、メタノール、
エタノールにやや可溶、水に不溶
（５）呈色反応：フォーリン反応陽性
（６）塩基性、中性、酸性の区別：酸性物質（７）色及
び形状：
白色結晶（融点１３６〜１３８℃）
上記の物理化学的性質及び構造解析結果から、単離され
る化合物は前記構造式で示されるイソインドリノン誘導
体であり、６− ［（２’Ｅ）−３°、７°−ジメチル
−５°−オキソ−２’、６ ’オクタジェニル］−７−
ヒドロキシ−５−メトキシ−Ｎ−（２”−フェニルエチ
ル）−１−イソインドリノンであることが決定された。8.08(s), 8.98(s), 7.20-7.
28(a)(4) Solubility in solvents: Soluble in ethyl acetate, chloroform, acetone, hexane, methanol,
Slightly soluble in ethanol, insoluble in water (5) Color reaction: Folin reaction positive (6) Basic, neutral, acidic distinction: Acidic substance (7) Color and shape: White crystals (melting point 136-138°C) From the above physicochemical properties and structural analysis results, the isolated compound is an isoindolinone derivative represented by the above structural formula, and is 6-[(2'E)-3°,7°-dimethyl-5°- oxo-2',6'octagenyl]-7-
It was determined to be hydroxy-5-methoxy-N-(2''-phenylethyl)-1-isoindolinone.
〈実施例〉
・イソインドリノン誘導体の抽出及び単離８５％エタノ
ール６文にヤマブシタケの朱子実体７．３ｋｇ￥を加え
、ホモジナイズ処理し、これを室温で一昼夜放置した後
、濾過して抽出液を得た。残渣に８５％エタノール４Ｌ
；Ｌを加え、同様に抽出処理を行なって抽出液を得、こ
れを１回目の抽出液と合わせた。そして合わせた抽出液
を減圧下に４０〜４５℃で加熱してエタノールを蒸発す
ることにより水相を得た。該水相にクロロホルム１文を
加え、振盪後、放置して分層したクロロホルム層を分取
した。残渣にクロロホルムｔｉを加え、同様に液液分配
抽出処理を行なってクロロホルム層を分取し、１回目の
クロロホルム層と合わせた０合わせたクロロホルム層を
減圧下に４０〜４５℃で加熱してクロロホルムを蒸発し
、更にデシケータで乾燥して、乾固物（Ａ）４．９９ｇ
を得た。<Example> ・Extraction and isolation of isoindolinone derivatives 7.3 kg of vermilion fruiting body of Yamabushitake was added to 6 volumes of 85% ethanol, homogenized, left overnight at room temperature, and then filtered to obtain the extract. Obtained. 4L of 85% ethanol to the residue
;L was added and the extraction process was performed in the same manner to obtain an extract, which was combined with the first extract. Then, the combined extracts were heated at 40 to 45° C. under reduced pressure to evaporate the ethanol, thereby obtaining an aqueous phase. One portion of chloroform was added to the aqueous phase, and after shaking, the mixture was left to stand and the separated chloroform layer was separated. Add chloroform ti to the residue, perform the same liquid-liquid partition extraction process to separate the chloroform layer, and combine with the first chloroform layer.The combined chloroform layer is heated at 40-45°C under reduced pressure to extract chloroform. was evaporated and further dried in a desiccator to obtain 4.99 g of dry matter (A).
I got it.
上記乾固物をヘキサンで溶解し、ワコーゲルＣ−２００
（和光紬薬社製）を用いてカラムクロマトグラフィーを
行なった。この際、展開溶媒として、順次極性が大きく
なるように、ヘキサン→クロロホルム呻りロロホルム／
アセトン（８／２）を各６０１用い、１０ｍ１の両分を
合計１８両分得た。このうちの第６及び７両分について
クロロホルム／ジエチルエーテル（７／３）で再結晶処
理を行ない、この際の結晶析出の時間的ズレにより合計
３グループを得、このうちの第１グループから前記構造
式で示されるイソインドリノン誘導体（Ｂ）３．２ｍｇ
を単離した。Dissolve the above dry matter in hexane and add Wakogel C-200.
Column chromatography was performed using (manufactured by Wako Tsumugi Pharmaceutical Co., Ltd.). At this time, as a developing solvent, hexane → chloroform, roloform/
Using 601 volumes of acetone (8/2) each, a total of 18 volumes of 10 ml were obtained. Of these, the 6th and 7th portions were recrystallized with chloroform/diethyl ether (7/3), and due to the time lag in crystal precipitation, a total of 3 groups were obtained. Isoindolinone derivative (B) represented by the structural formula 3.2 mg
was isolated.
Φ評価
粛代培養した子宮頚癌細胞を、細胞数が４×１０４／層
１となるように、牛胎児血清ｌＯ％を含むイーグルＭＥ
Ｍ培地で福釈して、懸濁液を調製し、該懸濁液をプラス
チック製９６六マイクロプレート（コーニング社製）の
各式にそれぞれ２００ｇ１注入した。これを５％炭酸ガ
ス培養器中で３７℃、２４時間培養後、この培養液中に
薬剤溶液をそれぞれｓｇ＋加え、更に上記と同様の条件
下で７２時間培養した。そして培地上清を除いた上で細
胞をメタノールで固定化し、ギムザ染色後、細胞増殖の
状態を鏡検した。薬剤溶液は、前記乾固物（Ａ）及び前
記イソインドリノン誘導体（Ｂ）をそれぞれ種々の濃度
となるようにメタノールに溶解して作製した。ΦEvaluation Subcultured cervical cancer cells were added to Eagle ME containing 10% fetal bovine serum so that the number of cells was 4 x 104/layer 1.
A suspension was prepared by enriching with M medium, and 200 g of the suspension was injected into each type of plastic 966 microplate (manufactured by Corning). After culturing this in a 5% carbon dioxide incubator at 37°C for 24 hours, each drug solution (sg+) was added to this culture solution, and the culture was further cultured for 72 hours under the same conditions as above. After removing the medium supernatant, the cells were fixed with methanol, and after Giemsa staining, the state of cell proliferation was examined under a microscope. The drug solution was prepared by dissolving the dried product (A) and the isoindolinone derivative (B) in methanol to various concentrations.
上記の鏡検下で殺細胞効果を調べ、生細胞数が全く認め
られない培地中の薬剤の最小濃度を最終有効濃度とした
。最終有効濃度は、乾固物（Ａ）の場合に１２５ ｇｇ
／ｍｌであり、またイソインドリノン誘導体（Ｂ）の
場合に６ 、３４ｇ ／ｍｌであったφ
〈発明の効果〉
以上説明した通りであるから、本発明に係る新規のイソ
インドリノン誘導体は子宮頚癌細胞に対して優れた殺細
胞効果を有し
しかもこれをヤマ
ブシタケから効率的に得ることができるという効果があ
る。The cell-killing effect was examined under the above-mentioned microscopic examination, and the minimum concentration of the drug in the medium at which no viable cells were observed was defined as the final effective concentration. The final effective concentration is 125 gg in the case of dry matter (A)
/ml, and in the case of isoindolinone derivative (B), it was 6.34 g /ml. <Effects of the Invention> As explained above, the novel isoindolinone derivative according to the present invention It has an excellent cell-killing effect on cervical cancer cells and can be efficiently obtained from Yamabushitake.
Claims (1)
とする子宮頚癌細胞の殺細胞剤。 ３、請求項２記載の殺細胞剤を製造するに際して、ヤマ
ブシタケの子実体を水及び有機溶媒の均一系で抽出処理
し、その抽出液から有機溶媒を蒸発して水相を得た後、
該水相を水及び有機溶媒の混合系で液液分配抽出処理し
て有機溶媒層を分取し、そして該有機溶媒層から有機溶
媒を蒸発することを特徴とする殺細胞剤の製造方法。 ４、水及び有機溶媒の混合系が水及びクロロホルムの混
合系である請求項３記載の殺細胞剤の製造方法。 ５、有機溶媒層から有機溶媒を蒸発した残りの乾固物を
クロマト分画処理し、更に再結晶処理する請求項３又は
４記載の殺細胞剤の製造方法。[Claims] 1. An isoindolinone derivative represented by the following structural formula. ▲There are mathematical formulas, chemical formulas, tables, etc.▼ 2. A cell-killing agent for cervical cancer cells containing the isoindolinone derivative according to claim 1 as an active ingredient. 3. When producing the cell killing agent according to claim 2, the fruiting body of Yamabushitake is extracted with a homogeneous system of water and an organic solvent, and the organic solvent is evaporated from the extract to obtain an aqueous phase.
A method for producing a cellicide, which comprises performing a liquid-liquid partition extraction treatment on the aqueous phase using a mixed system of water and an organic solvent to separate an organic solvent layer, and then evaporating the organic solvent from the organic solvent layer. 4. The method for producing a cell killing agent according to claim 3, wherein the mixed system of water and an organic solvent is a mixed system of water and chloroform. 5. The method for producing a cell-killing agent according to claim 3 or 4, wherein the dry matter remaining after evaporating the organic solvent from the organic solvent layer is subjected to chromatography fractionation treatment and further recrystallization treatment.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1294415A JP2757342B2 (en) | 1989-11-13 | 1989-11-13 | Isoindolinone derivative, cervical cancer cell killing agent containing the same as active ingredient, and method for producing the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1294415A JP2757342B2 (en) | 1989-11-13 | 1989-11-13 | Isoindolinone derivative, cervical cancer cell killing agent containing the same as active ingredient, and method for producing the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH03157367A true JPH03157367A (en) | 1991-07-05 |
| JP2757342B2 JP2757342B2 (en) | 1998-05-25 |
Family
ID=17807459
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1294415A Expired - Fee Related JP2757342B2 (en) | 1989-11-13 | 1989-11-13 | Isoindolinone derivative, cervical cancer cell killing agent containing the same as active ingredient, and method for producing the same |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2757342B2 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108440380A (en) * | 2018-03-27 | 2018-08-24 | 湖南新汇制药股份有限公司 | A kind of compound detached from hedgehog fungus mycelium |
| EP4342481A1 (en) * | 2022-09-26 | 2024-03-27 | Hifas da Terra SL | Use of an extract of hericium erinaceus for the treatment of vaginal and cervical diseases |
-
1989
- 1989-11-13 JP JP1294415A patent/JP2757342B2/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108440380A (en) * | 2018-03-27 | 2018-08-24 | 湖南新汇制药股份有限公司 | A kind of compound detached from hedgehog fungus mycelium |
| EP4342481A1 (en) * | 2022-09-26 | 2024-03-27 | Hifas da Terra SL | Use of an extract of hericium erinaceus for the treatment of vaginal and cervical diseases |
| WO2024068563A1 (en) * | 2022-09-26 | 2024-04-04 | Hifas Da Terra, S.L. | Use of an extract of hericium erinaceus for the treatment of vaginal and cervical diseases |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2757342B2 (en) | 1998-05-25 |
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