JP2623285B2 - Method for producing 2 (3-hydroxy-1,3,5 (10) -estrien-17-yl) propionic acid - Google Patents

Method for producing 2 (3-hydroxy-1,3,5 (10) -estrien-17-yl) propionic acid

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Publication number
JP2623285B2
JP2623285B2 JP63053453A JP5345388A JP2623285B2 JP 2623285 B2 JP2623285 B2 JP 2623285B2 JP 63053453 A JP63053453 A JP 63053453A JP 5345388 A JP5345388 A JP 5345388A JP 2623285 B2 JP2623285 B2 JP 2623285B2
Authority
JP
Japan
Prior art keywords
hydroxy
estrien
propionic acid
producing
substance
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 - Lifetime
Application number
JP63053453A
Other languages
Japanese (ja)
Other versions
JPH01229000A (en
Inventor
貢 飯田
誠 吉浜
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Snow Brand Milk Products Co Ltd
Original Assignee
Snow Brand Milk Products Co Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Snow Brand Milk Products Co Ltd filed Critical Snow Brand Milk Products Co Ltd
Priority to JP63053453A priority Critical patent/JP2623285B2/en
Publication of JPH01229000A publication Critical patent/JPH01229000A/en
Application granted granted Critical
Publication of JP2623285B2 publication Critical patent/JP2623285B2/en
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

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  • Steroid Compounds (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Description

【発明の詳細な説明】 (産業上の利用分野) 本発明は医薬、およびその中間体として有用な2(3
−ハイドロキシ−1,3,5(10)−エストリエン−17−イ
ル)プロピオン酸の製造法に関する。
DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention relates to pharmaceuticals and 2 (3) useful as intermediates thereof.
-Hydroxy-1,3,5 (10) -estrien-17-yl) propionic acid.

(従来の技術と発明が解決しようとする課題) 従来、種々のエストロジェン及びその誘導体が合成さ
れており、種々の生物学的活性を有することが知られて
いる。
(Prior Art and Problems to be Solved by the Invention) Conventionally, various estrogens and derivatives thereof have been synthesized and are known to have various biological activities.

特に医薬等において、生体内においてエストロジェン
活性を有する化合物については、その有用性から多くの
研究開発が行われてきた。
Particularly in pharmaceuticals and the like, many research and development have been conducted on compounds having estrogenic activity in vivo due to their usefulness.

これらの研究開発の一環として、本発明者らは、種々
のステロイド化合物の製造原料として知られているカン
ペステロール、コレステロール及びβ−シトステロール
の19位水酸化物を基質とし、これに、ロドコッカス・エ
クイの変異株であるK−3株を作用させると、新規なエ
ストロジェンである2(3−ハイドロキシ−1,3,5(1
0)−エストリエン−17−イル)プロピオン酸を産生す
ることを見いだし、本発明をなすに至った。
As a part of these research and development, the present inventors used campesterol, cholesterol and β-sitosterol, which are known as raw materials for production of various steroid compounds, as a substrate and provided a substrate for Rhodococcus equi. When the K-3 strain, which is a mutant strain of the present invention, is acted on, a new estrogen, 2 (3-hydroxy-1,3,5 (1
0) -Estrien-17-yl) propionic acid, which has led to the present invention.

従って、本発明は生物学的活性を有する新規なエスト
ロジェン、2(3−ハイドロキシ−1,3,5(10)−エス
トリエン−17−イル)プロピオン酸を産生することを課
題とする。
Therefore, an object of the present invention is to produce a novel estrogen having biological activity, 2 (3-hydroxy-1,3,5 (10) -estrien-17-yl) propionic acid.

(課題を解決するための手段) 本発明は、2(3−ハイドロキシ−1,3,5(10)−エ
ストリエン−17−イル)プロピオン酸として、次式: で表される物質を製造する方法であり、19位水酸基を有
するコレステロールを基質とし、ロドコッカス・エクイ
変異株K−3(FERM P9880)による微生物変換により得
ることを特徴とする。
(Means for Solving the Problems) The present invention provides 2 (3-hydroxy-1,3,5 (10) -estrien-17-yl) propionic acid represented by the following formula: Which is obtained by microbial conversion using Rhodococcus equi mutant K-3 (FERM P9880) using cholesterol having a hydroxyl group at position 19 as a substrate.

以下本発明を詳しく説明する。 Hereinafter, the present invention will be described in detail.

本物質は、生体内において顕著なエストロジェン活性
の生物学的活性を有する。
The substance has a remarkable estrogenic biological activity in vivo.

本物質は下記の理化学的性質を有することにより特定
される。
This substance is specified by having the following physicochemical properties.

外観 白色粉末 分子量 328 分子式 C21H28O3 EIマススペクトル m/z=342(メチル化物)(第1図) プロトン核磁気共鳴スペクトル(第
2図) 溶解性 エタノール、メタノール、酢酸エチル、クロロ
ホルムに可溶。水、ヘキサンに難溶。
Appearance White powder Molecular weight 328 Molecular formula C 21 H 28 O 3 EI mass spectrum m / z = 342 (methylated) (Fig. 1) Proton nuclear magnetic resonance spectrum (Fig. 2) Solubility Ethanol, methanol, ethyl acetate, chloroform soluble. Poorly soluble in water and hexane.

本発明にかかる2(3−ハイドロキシ−1,3,5(10)
−エストリエン−17−イル)プロピオン酸は、例えば下
記の方法により製造することが出来る。
2 (3-Hydroxy-1,3,5 (10) according to the present invention
-Estrien-17-yl) propionic acid can be produced, for example, by the following method.

ロドコッカス・エクイ変異株K−3株(微工研菌寄第
9880号、Rhodococcus equi MIL−1045)をコーンスティ
ーブリカー、酵母エキス等の炭素源、窒素源並びに無機
塩類を含有する培地中で培養、次いで培養終了後、19−
OHコレステロールまたは19−OHβ−シトステロール等の
19水酸化ステロイド化合物を上記培養液に添加して反応
を行う。反応終了後、培養液に硫酸などの酸を用いて、
培養液のpHを2付近まで低下させ、酢酸エチルにて抽出
を行う。得られた抽出液から溶媒成分を除去して粗精製
画分を得る。
Rhodococcus equi mutant K-3 strain
No. 9880, Rhodococcus equi MIL-1045) was cultured in a medium containing a carbon source such as corn steep liquor and yeast extract, a nitrogen source and inorganic salts.
OH cholesterol or 19-OHβ-sitosterol
A reaction is carried out by adding a 19-hydroxylated steroid compound to the above culture solution. After completion of the reaction, using an acid such as sulfuric acid in the culture solution,
The pH of the culture is lowered to around 2, and extraction is performed with ethyl acetate. The solvent component is removed from the obtained extract to obtain a crudely purified fraction.

この粗精製画分をシリカゲルカラム(Wako gel C−20
0)を用い、ヘキサン−酢酸エチルにて2(3−ハイド
ロキシ−1,3,5(10)−エストリエン−17−イル)プロ
ピオン酸を溶出して分取する。
This roughly purified fraction is applied to a silica gel column (Wako gel C-20).
Using (0), 2 (3-hydroxy-1,3,5 (10) -estrien-17-yl) propionic acid is eluted with hexane-ethyl acetate and fractionated.

なお、本物質と同時にエストロンが併産されるが、シ
リカゲルクロマトグラフィーでの溶出時間差に基づいて
分離、分取し得る。
Estrone is produced simultaneously with this substance, but can be separated and fractionated based on the elution time difference in silica gel chromatography.

次に本物質のエストロジェン活性を試験した結果を示
す。
Next, the results of testing the estrogenic activity of this substance are shown.

ラットを用いた膣脂垢法によった。 Vaginal staining using rats.

生後26〜27日の雌ラットにプロピレングリコールに溶
解した本物質10μgを3日間皮下注射した。4日目にこ
のラットより膣脂垢を採取し、検鏡したところ、有角上
皮ならびに角化細胞のみを認め、白血球の存在を認めな
かった。
26-27 days old female rats were injected subcutaneously with 10 μg of this substance dissolved in propylene glycol for 3 days. On the fourth day, vaginal dirt was collected from the rat and examined by microscopy. As a result, only corneal epithelium and keratinocytes were observed, and the presence of leukocytes was not observed.

従って、本物質はエストロジェンとしての生物学的活
性を有するものと判定された。
Therefore, this substance was determined to have biological activity as an estrogen.

(発明の効果) 本発明物質、2(3−ハイドロキシ−1,3,5(10)−
エストリエン−17−イル)プロピオン酸は、公知の物質
である19−OHコレステロール等を基質として特定な微生
物を利用して容易に製造する事ができ、また本物質はエ
ストロジェン活性を有するので医療分野での利用が期待
される。
(Effect of the Invention) The substance of the present invention, 2 (3-hydroxy-1,3,5 (10)-
Estrien-17-yl) propionic acid can be easily produced using a known microorganism such as 19-OH cholesterol as a substrate using a specific microorganism, and since this substance has estrogenic activity, it can be used in the medical field. The use of is expected.

(実施例) 以下に実施例により、本物質の製造法を具体的に説明
する。
(Examples) Hereinafter, the production method of the present substance will be specifically described with reference to Examples.

2(3−ハイドロキシ−1,3,5(10)−エストリエン−1
7−イル)プロピオン酸の製造法 ロドコッカス・エクイ変異株K−3株(微工研菌寄第
9880号、Rhodococcus equi MIL−1045)を表1に示した
組成のH.C.培地80mlに接種し、500mlの三角フラスコ中
で、27℃の温度にて22時間、振盪しながら(220r.p.
m.)培養した。
2 (3-hydroxy-1,3,5 (10) -estriene-1
Method for producing 7-yl) propionic acid Rhodococcus equi mutant K-3 strain
No. 9880, Rhodococcus equi MIL-1045) was inoculated into 80 ml of an HC medium having the composition shown in Table 1 and shaken (220 r.p. in a 500 ml Erlenmeyer flask) at a temperature of 27 ° C. for 22 hours.
m.) Cultured.

培養終了後、基質として19−OHコレステロールを上記
フラスコに8.5mg添加し、更に5時間上記と同様の条件
で反応を行った。
After completion of the culture, 8.5 mg of 19-OH cholesterol as a substrate was added to the flask, and the reaction was further performed under the same conditions as described above for 5 hours.

反応終了後、得られた培養液から酢酸エチルで3回抽
出を行い、抽出成分から、ロータリーエバポレーターに
より溶媒成分を除去した。
After completion of the reaction, the obtained culture solution was extracted three times with ethyl acetate, and the solvent component was removed from the extracted components by a rotary evaporator.

得られた粗精製画分を少量の酢酸エチルで溶解し、シ
リカゲルカラム(1cmφ×30cm)を用い、溶出溶媒とし
てヘキサン:酢酸エチル=8:2を用い、2(3−ハイド
ロキシ−1,3,5(10)−エストリエン−17−イル)プロ
ピオン酸を溶出、分取した。収量は6.16mg(72.5%)で
あった。
The obtained crude purified fraction was dissolved in a small amount of ethyl acetate, and using a silica gel column (1 cmφ × 30 cm), hexane: ethyl acetate = 8: 2 as an elution solvent, and 2 (3-hydroxy-1,3,3 5 (10) -Estrien-17-yl) propionic acid was eluted and fractionated. The yield was 6.16 mg (72.5%).

得られた物質の理化学的性質は、前述の通りであっ
た。
The physicochemical properties of the obtained substance were as described above.

表1 H.C.培地 Na2HPO4・12H2O 6.0g KH2PO4 3.0g NH4Cl 1.0g MgSO4・7H2O 0.2g NaCl 0.5g 酵母エキス 0.1g コーンスティープリカー 5.0g 精製水 1000 ml pH 7.2Table 1 HC medium Na 2 HPO 4・ 12H 2 O 6.0g KH 2 PO 4 3.0g NH 4 Cl 1.0g MgSO 4・ 7H 2 O 0.2g NaCl 0.5g Yeast extract 0.1g Corn steep liquor 5.0g Purified water 1000 ml pH 7.2

【図面の簡単な説明】[Brief description of the drawings]

第1図は、本発明実施例による2(3−ハイドロキシ−
1,3,5(10)−エストリエン−17−イル)プロピオン酸
のEIマススペクトル(m/z=342(メチル化物))を示す
図、 第2図は、前記化合物のプロトン核磁気共鳴スペクトル
を示す図である。
FIG. 1 shows 2 (3-hydroxy-) according to an embodiment of the present invention.
FIG. 2 shows an EI mass spectrum (m / z = 342 (methylated)) of 1,3,5 (10) -estrien-17-yl) propionic acid. FIG. 2 shows a proton nuclear magnetic resonance spectrum of the compound. FIG.

Claims (1)

(57)【特許請求の範囲】(57) [Claims] 【請求項1】19位水酸基を有するコレステロールを基質
とし、ロドコッカス・エクイ変異株K−3(FERM P−98
80)による微生物変換により得ることを特徴とする、次
式: で表される2(3−ハイドロキシ−1,3,5(10)−エス
トリエン−17−イル)プロピオン酸の製造法。
A cholesterol having a hydroxyl group at the 19-position is used as a substrate, and Rhodococcus equi mutant K-3 (FERM P-98) is used.
80) characterized by being obtained by microbial conversion according to the following formula: A method for producing 2 (3-hydroxy-1,3,5 (10) -estrien-17-yl) propionic acid represented by the formula:
JP63053453A 1988-03-09 1988-03-09 Method for producing 2 (3-hydroxy-1,3,5 (10) -estrien-17-yl) propionic acid Expired - Lifetime JP2623285B2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP63053453A JP2623285B2 (en) 1988-03-09 1988-03-09 Method for producing 2 (3-hydroxy-1,3,5 (10) -estrien-17-yl) propionic acid

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP63053453A JP2623285B2 (en) 1988-03-09 1988-03-09 Method for producing 2 (3-hydroxy-1,3,5 (10) -estrien-17-yl) propionic acid

Publications (2)

Publication Number Publication Date
JPH01229000A JPH01229000A (en) 1989-09-12
JP2623285B2 true JP2623285B2 (en) 1997-06-25

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ID=12943275

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Country Status (1)

Country Link
JP (1) JP2623285B2 (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB9910934D0 (en) * 1999-05-11 1999-07-14 Res Inst Medicine Chem Chemical compounds
CN109423455B (en) * 2017-08-30 2022-08-19 中国石油化工股份有限公司 Rhodococcus equi, and identification method and application thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BIOCHEMISTRY=1968 *
J.AM.CHEM.SOC=1967 *

Also Published As

Publication number Publication date
JPH01229000A (en) 1989-09-12

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