JP2923692B2 - Enzymatic resolution method - Google Patents
Enzymatic resolution methodInfo
- Publication number
- JP2923692B2 JP2923692B2 JP33572190A JP33572190A JP2923692B2 JP 2923692 B2 JP2923692 B2 JP 2923692B2 JP 33572190 A JP33572190 A JP 33572190A JP 33572190 A JP33572190 A JP 33572190A JP 2923692 B2 JP2923692 B2 JP 2923692B2
- Authority
- JP
- Japan
- Prior art keywords
- compound
- ethyl
- pyrano
- indolizine
- ethylenedioxy
- 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.)
- Expired - Fee Related
Links
Description
【発明の詳細な説明】 〔産業上の利用分野〕 本発明は酵素を利用した光学分割法に関し、更に詳細
には合成抗癌剤として有用であるカンプトテシン誘導体
の重要な合成中間体である次式（１） で表わされる（Ｓ）−４−アシルオキシ−４−エチル−
6,6−（エチレンジオキシ）−7,8−ジヒドロ−1H−ピラ
ノ〔3,4−ｆ〕インドリジン−3,10（4H）−ジオン酵素
を利用した光学分割により製造する方法に関する。Description: TECHNICAL FIELD The present invention relates to an optical resolution method using an enzyme, and more particularly, to the following formula (1) which is an important synthetic intermediate of a camptothecin derivative useful as a synthetic anticancer agent. ) (S) -4-acyloxy-4-ethyl- represented by
The present invention relates to a method for producing a compound by optical resolution using an enzyme of 6,6- (ethylenedioxy) -7,8-dihydro-1H-pyrano [3,4-f] indolizine-3,10 (4H) -dione.
従来、次の式（３） で表わされるカンプトテシン誘導体は、合成抗癌剤とし
て有用であり、その合成中間体として下記化合物（４）
が重要であることが知られている（特開昭63−152382号
公報）。Conventionally, the following equation (3) Is useful as a synthetic anticancer agent, and as a synthetic intermediate thereof, the following compound (4)
Is known to be important (JP-A-63-152382).
かかる化合物（４）の製造法としては、当該化合物
（４）のラセミ体をジアステレオマーに誘導後分別結晶
化による光学分割法（特開昭63−152382号公報）等が知
られている。 As a method for producing such a compound (4), an optical resolution method by inducing a racemate of the compound (4) into a diastereomer and then fractional crystallization (JP-A-63-152382) is known.
しかしながら、これら従来の光学分割法は操作が煩雑
である、生成物の収率が低い等の欠点を有し、工業的な
製法としては充分満足できるものではなかった。However, these conventional optical resolution methods have disadvantages such as complicated operation and low product yield, and have not been sufficiently satisfactory as an industrial production method.
かかる実状において、本発明者らは酵素を利用して化
合物（４）のみを選択的に得るべく種々探索したとこ
ろ、パパイン又はフィシンはラセミ体である下記化合物
（２）のＲ体にのみ作用してアシル基を除去し、Ｓ体に
は作用しないため、化合物（４）へ容易に導くことので
きる化合物（１）が選択的に得られることを見出し、本
発明を完成するに到った。In this situation, the present inventors have conducted various searches for selectively obtaining only compound (4) using an enzyme, and found that papain or ficin acts only on the R-form of the following racemic compound (2). It has been found that the compound (1), which can be easily led to the compound (4), can be selectively obtained because the acyl group is removed and the compound (1) does not act on the S-isomer, thereby completing the present invention.
本発明は次の反応式によって示される。 The present invention is illustrated by the following reaction scheme.
（式中、Ｒはアルキル基を示す） すなわち、一般式（２）で表わされる４−アシルオキ
シ−４−エチル−6,6−（エチレンジオキシ）−7,8−ジ
ヒドロ−1H−ピラノ〔3,4−ｆ〕インドリジン−3,10（4
H）−ジオンのラセミ体に、パパイン又はフィシンを作
用させることを特徴とする化合物（１）の製造法であ
る。 (Wherein, R represents an alkyl group) That is, 4-acyloxy-4-ethyl-6,6- (ethylenedioxy) -7,8-dihydro-1H-pyrano [3 , 4-f] indolizine-3,10 (4
A method for producing a compound (1), which comprises reacting papain or ficin with a racemic form of H) -dione.
本発明方法の原料化合物を表わす上記一般式（２）
中、Ｒで示されるアルキル基としてはメチル基、エチル
基、ｎ−プロピル基等の低級アルキル基が好ましい。The above general formula (2) representing a starting compound of the method of the present invention.
Among them, the alkyl group represented by R is preferably a lower alkyl group such as a methyl group, an ethyl group, and an n-propyl group.
かかる化合物（２）のラセミ体（以下、単に化合物
（２）という）は、例えば特開昭63−152382号記載の方
法に従って得られた４−エチル−6,6−（エチレンジオ
キシ）−7,8−ジヒドロ−４−ヒドロキシ−1H−ピラノ
〔3,4−ｆ〕インドリジン−3,10（4H）−ジオンに、脂
肪酸無水物、脂肪酸ハライド等の脂肪酸の反応性誘導体
を反応させることにより製造される。Such a racemic compound (2) (hereinafter simply referred to as compound (2)) can be obtained, for example, from 4-ethyl-6,6- (ethylenedioxy) -7 obtained by the method described in JP-A-63-152382. , 8-dihydro-4-hydroxy-1H-pyrano [3,4-f] indolizine-3,10 (4H) -dione is reacted with a reactive derivative of a fatty acid such as a fatty acid anhydride or a fatty acid halide. Manufactured.
本発明方法を実施するには、まず化合物（２）を水又
は水と酢酸エチル、イソプロピルエーテル、ｎ−ヘキサ
ン、ベンゼン、トルエンの如き有機溶媒との混合溶媒に
懸濁し、次いでパパイン又はフィシンを静かに加え撹拌
すればよい。In order to carry out the method of the present invention, the compound (2) is first suspended in water or a mixed solvent of water and an organic solvent such as ethyl acetate, isopropyl ether, n-hexane, benzene and toluene, and then papain or ficin is gently suspended. And stirring.
反応は、通常５〜60℃、好ましくは30〜50℃、pH4〜
９好ましくは、６〜７で10〜48時間程度行われる。反応
に使用する化合物（２）の濃度は、0.1〜５重量％の間
で可能であるが通常0.5〜２重量％程度で行うのが望ま
しい。パパイン又はフィシンの使用量は特に限定されな
いが、化合物（２）に対し１重量倍から10重量倍用いる
のが適当である。反応終了後、反応液を濾過、減圧濃
縮、カラムクロマトグラフィー等の手段を用いて化合物
（１）を単離することができる。The reaction is usually performed at 5 to 60 ° C, preferably 30 to 50 ° C, pH 4 to
9 Preferably, it is performed for 6 to 7 for about 10 to 48 hours. The concentration of the compound (2) used in the reaction can be between 0.1 and 5% by weight, but it is usually desirable to carry out the reaction at about 0.5 to 2% by weight. The amount of papain or ficin to be used is not particularly limited, but it is appropriate to use 1 to 10 times by weight the compound (2). After the completion of the reaction, the compound (1) can be isolated using a means such as filtration, concentration under reduced pressure, and column chromatography.
得られた化合物（１）は、脱アシル化して化合物
（４）へと導き、次いで例えば特開昭63−152382号公報
記載の方法により前記カンプトテシン誘導体（３）、そ
の他の合成抗癌剤に導くことができる。The obtained compound (1) can be deacylated to obtain a compound (4), and then can be introduced into the camptothecin derivative (3) and other synthetic anticancer agents by the method described in, for example, JP-A-63-152382. it can.
本発明の製造法は温和な条件下で反応を行うことがで
き、反応における副反応がほとんど無いため目的の化合
物（１）を高純度に製造することができる。従って本発
明の製造法は、化合物（１）の工業的製造方法として優
れたものである。According to the production method of the present invention, the reaction can be carried out under mild conditions, and since there are almost no side reactions in the reaction, the target compound (1) can be produced with high purity. Therefore, the production method of the present invention is excellent as an industrial production method of compound (1).
以下、本発明を更に実施例により説明するが、本発明
はこれにより限定されるものではない。Hereinafter, the present invention will be further described with reference to examples, but the present invention is not limited thereto.
参考例１ ４−アセトキシ−４−エチル−6,6−（エチレンジオキ
シ）−7,8−ジヒドロ−1H−ピラノ（3,4−ｆ）インドリ
ジン−3,10（4H）−ジオン ４−エチル−6,6−（エチレンジオキシ）−7,8−ジヒ
ドロ−４−ヒドロキシ−1H−ピラノ〔3,4−ｆ〕インド
リジン−3,10（4H）−ジオン3.07gを塩化メチレン10ml
に懸濁し氷冷撹拌下、ジメチルアミノピリジン124mg、
無水酢酸1.13mlを加え室温で４時間放置した。HPLCで反
応の終了を確認後、反応液を減圧下濃縮した。残渣をイ
ソプロピルアルコール20mlから結晶化し濾過した。結晶
を冷イソプロピルアルコールで洗浄後乾燥し標記化合物
3.41gを得た。Reference Example 1 4-acetoxy-4-ethyl-6,6- (ethylenedioxy) -7,8-dihydro-1H-pyrano (3,4-f) indolizine-3,10 (4H) -dione 4- 3.07 g of ethyl-6,6- (ethylenedioxy) -7,8-dihydro-4-hydroxy-1H-pyrano [3,4-f] indolizine-3,10 (4H) -dione and 10 ml of methylene chloride
Suspended under ice-cooling and stirring, dimethylaminopyridine 124 mg,
1.13 ml of acetic anhydride was added and left at room temperature for 4 hours. After confirming the completion of the reaction by HPLC, the reaction solution was concentrated under reduced pressure. The residue was crystallized from 20 ml of isopropyl alcohol and filtered. The crystals were washed with cold isopropyl alcohol and dried to give the title compound.
3.41 g was obtained.
IRνmaxcm-11740,1660,1610（Ｃ＝０） FT−NMR（CDCl3中のδ値ppm） 0.87（3H,t,J＝7Hz） 2.07（2H,t,J＝7Hz） 2.11（3H,s） 2.42（2H,t,J＝7Hz） 4.0〜4.3（6H,m） 5.36（2H,ABquartet,J＝17Hz,45Hz） 6.07（1H,s） 実施例１ （Ｓ）−４−アセトキシ−４−エチル−6,6−（エチレ
ンジオキシ）−7,8−ジヒドロ−1H−ピラノ〔3,4−ｆ〕
インドリジン−3,10（4H）−ジオン ４−アセトキシ−４−エチル−6,6−（エチレンジオ
キシ）−7,8−ジヒドロ−1H−ピラノ〔3,4−ｆ〕インド
リジン−3,10（4H）−ジオン1.50gを水：酢酸エチル（9
0:10）の混合溶媒（含0.1Mリン酸緩衝液pH6.0）150mlに
懸濁し、次いでパパイン（メルク社製）7.5g、２−メル
カプトエタノール0.6mlを加え40℃で24時間撹拌した。H
PLCで反応の終了を確認後、反応液を分液（水−クロロ
ホルム）し生成物を抽出した（50ml×３）。有機層を水
で洗浄後乾燥し濃縮した。残渣をシリカゲルクロマトグ
ラフィー（クロロホルム−アセトン）で精製し、705mg
の標題化合物と594mgの（Ｒ）−４−エチル−6,6−（エ
チレンジオキシ）−7,8−ジヒドロ−４−ヒドロキシ−1
H−ピラノ〔3,4−ｆ〕インドリジン−3,10（4H）−ジオ
ンを回収した。IRν max cm -1 1740,1660,1610 (C = 0) FT-NMR (δ value ppm in CDCl 3 ) 0.87 (3H, t, J = 7 Hz) 2.07 (2H, t, J = 7 Hz) 2.11 (3H , s) 2.42 (2H, t, J = 7Hz) 4.0-4.3 (6H, m) 5.36 (2H, ABquartet, J = 17Hz, 45Hz) 6.07 (1H, s) Example 1 (S) -4-acetoxy- 4-ethyl-6,6- (ethylenedioxy) -7,8-dihydro-1H-pyrano [3,4-f]
Indolizine-3,10 (4H) -dione 4-acetoxy-4-ethyl-6,6- (ethylenedioxy) -7,8-dihydro-1H-pyrano [3,4-f] indolizine-3, 1.50 g of 10 (4H) -dione in water: ethyl acetate (9
0:10) in 150 ml of a mixed solvent (containing 0.1 M phosphate buffer, pH 6.0), 7.5 g of papain (manufactured by Merck) and 0.6 ml of 2-mercaptoethanol were added, and the mixture was stirred at 40 ° C. for 24 hours. H
After confirming the completion of the reaction by PLC, the reaction solution was separated (water-chloroform) and the product was extracted (50 ml × 3). The organic layer was washed with water, dried and concentrated. The residue was purified by silica gel chromatography (chloroform-acetone) to give 705 mg.
594 mg of (R) -4-ethyl-6,6- (ethylenedioxy) -7,8-dihydro-4-hydroxy-1
H-pyrano [3,4-f] indolizine-3,10 (4H) -dione was recovered.
得られた目的化合物の機器分析値は別途合成のそれと
完全に一致し光学純度は、光学異性体分離カラムによる
分析により99％e.e.以上であった。The instrumental analysis value of the obtained target compound completely coincided with that of the separately synthesized compound, and the optical purity was 99% ee or more as analyzed by an optical isomer separation column.
IRνmaxcm-11740,1660,1610（Ｃ＝０） FT−NMR（CDCl3中のδ値ppm） 0.87（3H,t,J＝7Hz） 2.07（2H,t,J＝7Hz） 2.11（3H,s） 2.42（2H,t,J＝7Hz） 4.0〜4.3（6H,m） 5.36（2H,ABquartet,J＝17Hz,45Hz） 6.07（1H,s） HPLC条件 カラム:ULTRON ES−OVM（信和化工社製） 移動相:2.5％エタノール（含20mMリン酸緩衝液pH6.
0） 流 速:1.0ml/min 波 長:254nm 実施例２ （Ｓ）−４−アセトキシ−４−エチル−6,6−（エチレ
ンジオキシ）−7,8−ジヒドロ−1H−ピラノ〔3,4−ｆ〕
インドリジン−3,10（4H）−ジオン ４−アセトキシ−４−エチル−6,6−（エチレンジオ
キシ）−7,8−ジヒドロ−1H−ピラノ〔3,4−ｆ〕インド
リジン−3,10（4H）−ジオン750mgを0.1Mリン酸緩衝液
（pH6.0）150mlに懸濁し、次いでフィシン（長瀬産業株
式会社製）7.5g、２−メルカプトエタノール0.6mlを加
え40℃で48時間撹拌した。HPLCで反応の終了を確認後、
反応液を分液（水−クロロホルム）し生成物を抽出した
（25ml×３）。有機層を水で洗浄後乾燥し濃縮した後、
残渣をシリカゲルクロマトグラフィー（クロロホルム−
アセトン）で精製し、360mgの標題化合物と290mgの
（Ｒ）−４−エチル−6,6−（エチレンジオキシ）−7,8
−ジヒドロ−４−ヒドロキシ−1H−ピラノ〔3,4−ｆ〕
インドリジン−3,10（4H）−ジオンを回収した。IRν max cm -1 1740,1660,1610 (C = 0) FT-NMR (δ value ppm in CDCl 3 ) 0.87 (3H, t, J = 7 Hz) 2.07 (2H, t, J = 7 Hz) 2.11 (3H , s) 2.42 (2H, t, J = 7Hz) 4.0 to 4.3 (6H, m) 5.36 (2H, ABquartet, J = 17Hz, 45Hz) 6.07 (1H, s) HPLC conditions Column: ULTRON ES-OVM (Shinwa Chemical Industries, Ltd.) Mobile phase: 2.5% ethanol (20 mM phosphate buffer pH 6.
0) Flow rate: 1.0 ml / min Wavelength: 254 nm Example 2 (S) -4-acetoxy-4-ethyl-6,6- (ethylenedioxy) -7,8-dihydro-1H-pyrano [3, 4-f]
Indolizine-3,10 (4H) -dione 4-acetoxy-4-ethyl-6,6- (ethylenedioxy) -7,8-dihydro-1H-pyrano [3,4-f] indolizine-3, 750 mg of 10 (4H) -dione was suspended in 150 ml of 0.1 M phosphate buffer (pH 6.0), and then 7.5 g of ficin (manufactured by Nagase & Co., Ltd.) and 0.6 ml of 2-mercaptoethanol were added, followed by stirring at 40 ° C. for 48 hours. did. After confirming the completion of the reaction by HPLC,
The reaction solution was separated (water-chloroform) to extract the product (25 ml × 3). After washing the organic layer with water, drying and concentrating,
The residue is purified by silica gel chromatography (chloroform-
(Acetone) to give 360 mg of the title compound and 290 mg of (R) -4-ethyl-6,6- (ethylenedioxy) -7,8.
-Dihydro-4-hydroxy-1H-pyrano [3,4-f]
Indolizine-3,10 (4H) -dione was recovered.
得られた目的化合物の光学純度は、光学異性体分離カ
ラムによる分析により96％e.e.であった。The optical purity of the obtained target compound was 96% ee by analysis with an optical isomer separation column.
───────────────────────────────────────────────────── フロントページの続き (58)調査した分野(Int.Cl.6，ＤＢ名) C12P 41/00 ＲＥＧＩＳＴＲＹ（ＳＴＮ) ＣＡ（ＳＴＮ)────────────────────────────────────────────────── ─── Continued on the front page (58) Field surveyed (Int. Cl. 6 , DB name) C12P 41/00 REGISTRY (STN) CA (STN)
Claims (1)
（エチレンジオキシ）−7,8−ジヒドロ−1H−ピラノ
〔3,4−ｆ〕インドリジン−3,10（4H）−ジオンのラセ
ミ体に、パパイン又はフィシンを作用させることを特徴
とする、次式（１） で表わされる（Ｓ）−４−アシルオキシ−４−エチル−
6,6−（エチレンジオキシ）−7,8−ジヒドロ−1H−ピラ
ノ〔3,4−ｆ〕インドリジン−3,10（4H）−ジオンの製
造法。1. The general formula (2) (Wherein R represents an alkyl group) 4-acyloxy-4-ethyl-6,6-
Reacting a racemate of (ethylenedioxy) -7,8-dihydro-1H-pyrano [3,4-f] indolizine-3,10 (4H) -dione with papain or ficin. The following equation (1) (S) -4-acyloxy-4-ethyl- represented by
A method for producing 6,6- (ethylenedioxy) -7,8-dihydro-1H-pyrano [3,4-f] indolizine-3,10 (4H) -dione.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP33572190A JP2923692B2 (en) | 1990-11-30 | 1990-11-30 | Enzymatic resolution method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP33572190A JP2923692B2 (en) | 1990-11-30 | 1990-11-30 | Enzymatic resolution method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH04200392A JPH04200392A (en) | 1992-07-21 |
| JP2923692B2 true JP2923692B2 (en) | 1999-07-26 |
Family
ID=18291729
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP33572190A Expired - Fee Related JP2923692B2 (en) | 1990-11-30 | 1990-11-30 | Enzymatic resolution method |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2923692B2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SG104284A1 (en) * | 1996-10-30 | 2004-06-21 | Tanabe Seiyaku Co | S type 2-substituted hydroxy-2-indolidinylbutyric ester compounds and process for preparation thereof |
-
1990
- 1990-11-30 JP JP33572190A patent/JP2923692B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JPH04200392A (en) | 1992-07-21 |
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