JPH0889277A - Production of 12-hydroxy-6-o-alkylerythromycin and its homologue - Google Patents
Production of 12-hydroxy-6-o-alkylerythromycin and its homologueInfo
- Publication number
- JPH0889277A JPH0889277A JP6232558A JP23255894A JPH0889277A JP H0889277 A JPH0889277 A JP H0889277A JP 6232558 A JP6232558 A JP 6232558A JP 23255894 A JP23255894 A JP 23255894A JP H0889277 A JPH0889277 A JP H0889277A
- Authority
- JP
- Japan
- Prior art keywords
- hydrogen atom
- alkylerythromycin
- analogs
- methylerythromycin
- hydroxy
- 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
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- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims abstract 4
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- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- GDSRMADSINPKSL-HSEONFRVSA-N gamma-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO GDSRMADSINPKSL-HSEONFRVSA-N 0.000 description 1
- 229940080345 gamma-cyclodextrin Drugs 0.000 description 1
- 235000021312 gluten Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- RUTXIHLAWFEWGM-UHFFFAOYSA-H iron(3+) sulfate Chemical compound [Fe+3].[Fe+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O RUTXIHLAWFEWGM-UHFFFAOYSA-H 0.000 description 1
- 229910000360 iron(III) sulfate Inorganic materials 0.000 description 1
- 239000003120 macrolide antibiotic agent Substances 0.000 description 1
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 1
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229960003966 nicotinamide Drugs 0.000 description 1
- 235000005152 nicotinamide Nutrition 0.000 description 1
- 239000011570 nicotinamide Substances 0.000 description 1
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 125000000962 organic group Chemical group 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000004810 partition chromatography Methods 0.000 description 1
- 235000021317 phosphate Nutrition 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000012064 sodium phosphate buffer Substances 0.000 description 1
- 229940074404 sodium succinate Drugs 0.000 description 1
- ZDQYSKICYIVCPN-UHFFFAOYSA-L sodium succinate (anhydrous) Chemical compound [Na+].[Na+].[O-]C(=O)CCC([O-])=O ZDQYSKICYIVCPN-UHFFFAOYSA-L 0.000 description 1
- 239000004328 sodium tetraborate Substances 0.000 description 1
- XBYBXDBJLZDASJ-UHFFFAOYSA-M sodium;dimethylarsinate;hydrochloride Chemical compound [Na+].Cl.C[As](C)([O-])=O XBYBXDBJLZDASJ-UHFFFAOYSA-M 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000012134 supernatant fraction Substances 0.000 description 1
- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
Abstract
Description
【0001】[0001]
【産業上の利用分野】本発明は12−ヒドロキシ−6−
O−メチルエリスロマイシン及びその類縁体を微生物を
利用して製造する方法に関する。The present invention relates to 12-hydroxy-6-
The present invention relates to a method for producing O -methylerythromycin and its analogs using a microorganism.
【0002】[0002]
【従来の技術】有機化学的方法により、6−O−アルキ
ルエリスロマイシン及びその類縁体の12位の水素原子
を直接水酸化することは極めて困難で、その例は今だ報
告されていない。また、微生物を用いた酵素化学的方法
により、6−O−アルキルエリスロマイシン及びその類
縁体の12位の水素原子を直接水酸化する方法もこれま
でに報告されていない。2. Description of the Related Art It is extremely difficult to directly hydroxylate the hydrogen atom at the 12-position of 6- O- alkylerythromycin and its analogs by an organic chemical method, and an example thereof has not been reported yet. Further, a method of directly hydroxylating the 12-position hydrogen atom of 6- O- alkylerythromycin and its analogs by an enzymatic chemical method using a microorganism has not been reported so far.
【0003】[0003]
【発明が解決しようとする課題】本発明は、より容易な
手段による12−ヒドロキシ−6−O−メチルエリスロ
マイシン及びその類縁体を得る方法を提供することを目
的とする。The object of the present invention is to provide a method for obtaining 12-hydroxy-6- O -methylerythromycin and its analogs by an easier means.
【0004】[0004]
【課題を解決するための手段】本発明者は、特定の微生
物を利用することにより、6−O−アルキルエリスロマ
イシン及びその類縁体の12位の水素原子を直接水酸化
できることを見いだし、本発明を完成した。本発明は、
6−O−アルキルエリスロマイシン及びその類縁体を水
酸化する放線菌菌体又はその菌体が産生する酵素を含有
する溶液中に、12位に水素原子を有する6−O−アル
キルエリスロマイシン及びその類縁体を加えてその水素
原子を水酸基に変換することを特徴とする12−ヒドロ
キシ−6−O−アルキルエリスロマイシン及びその類縁
体の製造方法である。The present inventor has found that the hydrogen atom at the 12-position of 6- O- alkylerythromycin and its analogs can be directly hydroxylated by utilizing a specific microorganism, and the present invention has been made. completed. The present invention
6- O- Alkylerythromycin and its analogs having a hydrogen atom at the 12-position in a solution containing actinomycetes or hydroxylated hydroxylated 6- O- alkylerythromycin and its analogs Is added to convert the hydrogen atom to a hydroxyl group, which is a method for producing 12-hydroxy-6- O- alkylerythromycin and its analogs.
【0005】本発明の製造方法は、12位に水素原子を
有する6−O−アルキルエリスロマイシン及びその類縁
体の12位を直接、1工程で水酸化する方法であり、1
2位以外にいずれの置換基を有していてもよい。従っ
て、本発明における12位に水素原子を有する6−O−
アルキルエリスロマイシン(以下、単に基質ということ
がある)とは、たとえば、6−O−メチルエリスロマイ
シンB、6−O−エチルエリスロマイシンB、6−O−
プロピルエリスロマイシンB、6−O−ブチルエリスロ
マイシンBなどである。また、12位に水素原子を有す
る6−O−アルキルエリスロマイシンの類縁体(以下、
単に基質ということがある)とは、たとえば、6−O−
メチルエリスロマイシンBオキシム、デクラジノシル−
6−O−メチルエリスロマイシンB、14−ヒドロキシ
−6−O−メチルエリスロマイシンB、6,11−ジ−
O−メチルエリスロマイシンB、6,4”−ジ−O−メ
チルエリスロマイシンB、6,11,4”−トリ−O−
メチルエリスロマイシンB、6−O−メチルエリスロノ
ライドBなどである。The production method of the present invention is a method of directly hydroxylating the 12-position of 6- O- alkylerythromycin and its analogs having a hydrogen atom at the 12-position in one step.
It may have any substituent other than the 2-position. Therefore, 6- O- having a hydrogen atom at the 12-position in the present invention.
Alkylerythromycin (hereinafter sometimes simply referred to as a substrate) means, for example, 6- O -methylerythromycin B, 6- O -ethylerythromycin B, 6- O-.
Examples include propylerythromycin B and 6- O -butylerythromycin B. Further, an analog of 6- O- alkylerythromycin having a hydrogen atom at the 12-position (hereinafter,
Sometimes referred to as a substrate), for example, 6- O-
Methylerythromycin B oxime, decladinosyl-
6- O -methylerythromycin B, 14-hydroxy-6- O -methylerythromycin B, 6,11-di-
O -methylerythromycin B, 6,4 "-di- O -methylerythromycin B, 6,11,4" -tri- O-
Methylerythromycin B, 6- O -methylerythronolide B and the like.
【0006】本発明に使用される放線菌とは、6−O−
メチルエリスロマイシン及びその類縁体の12位の水素
原子を水酸化する能力を有する菌である。それらは例え
ば、バージェーズ・マニュアル・オブ・システマティッ
ク・バクテリオロジー(Bergey's Manual of Systemati
c Bacteriology),第4巻,1989年に記載されてい
る放線菌であり、代表的にはアクチノマジュラ(Actino
madura)属、アミコラタ(Amycolata)属、ノカルジア
(Nocardia)属、ミクロモノスポラ(Micromonospora)
属、サッカロポリスポラ(Saccharopolyspora)属、ス
トレプトミセス(Streptomyces)属などを挙げることが
でき、好ましくはミクロモノスポラ属、ストレプトミセ
ス属、サッカロポリスポラ属に属する放線菌であり、最
も好ましくはミクロモノスポラ・メガロミセア亜種ニグ
ラ(Micromonospora megalomicea subsp. nigra) I
FO14114、ストレプトミセス・フェレウス(Stre
ptomyces felleus) JCM4031、サッカロポリス
ポラ・エリスレア(Saccharopolyspora erythraea)
IFO13426である。The actinomycetes used in the present invention are 6- O-
It is a bacterium having the ability to hydroxylate the hydrogen atom at the 12-position of methylerythromycin and its analogs. They are, for example, Bergey's Manual of Systemati
c Bacteriology), Vol. 4, 1989, typically Actinomajura ( Actino).
madura) genus, Amikorata (Amycolata) genus Nocardia (Nocardia) sp., Micromonospora (Micromonospora)
Genus, Saccharopolyspora ( Saccharopolyspora ) genus, Streptomyces ( Streptomyces ) genus and the like can be mentioned, preferably Micromonospora, Streptomyces, Saccharopolyspora actinomycetes, most preferably Micromonospora megalomicea subsp. Nigra I
FO14114, Streptomyces ferreus ( Stre
ptomyces felleus ) JCM4031, Saccharopolyspora erythraea
IFO 13426.
【0007】ミクロモノスポラ・メガロミセア亜種ニグ
ラ IFO14114(「Agricultural Research Cult
ure Collection」の保存株NRRL3275と起源を同
じくする菌株;文献:米国特許第3,632,750
号)及びサッカロポリスポラ・エリスレア IFO13
426[「Agricultural Research Culture Collectio
n」の保存株NRRL2338と起源を同じくする菌
株;文献:インターナショナル・ジャーナル・オブ・シ
ステマティック・バクテリオロジー(Int. J. Syst.Bac
teriol.),第37巻,458頁,1987年及びイン
ターナショナル・ジャーナル・オブ・システマティック
・バクテリオロジー(Int. J. Syst. Bacteriol.),第
37巻,19〜22頁,1987年]はいずれも財団法
人発酵研究所の保存株であり、それぞれ「FERM−P
*****」及び「FERM−P*****」として工
業技術院生命工学工業研究所に寄託されており、これら
の菌株の菌学的性状は前記文献により明かにされてい
る。ストレプトミセス・フェレウスJCM4031
(「Agricultural Research Culture Collection」の保
存株NRRL2251と起源を同じくする菌株;文献:
米国特許第2,693,433号)は理化学研究所微生
物系統保存施設の保存株であり、FERM−P****
*として工業技術院生命工学工業研究所に寄託されてお
り、前記文献により本菌株の菌学的性状は明かにされて
いる。Micromonospora megalomicea subsp. Nigra IFO14114 ("Agricultural Research Cult
Strains of the same origin as the preserved strain NRRL3275 of the "ure Collection"; Reference: US Pat. No. 3,632,750
No.) and Saccharopoli spora erythrea IFO13
426 ["Agricultural Research Culture Collectio
Strain that has the same origin as the "n" preserved strain NRRL2338; Reference: International Journal of Systematic Bacteriology (Int. J. Syst. Bac
teriol.), 37, 458, 1987, and International Journal of Systematic Bacteriology (Int. J. Syst. Bacteriol.), 37, 19-22, 1987]. Preserved strains of the Fermentation Research Institute, each of which is called "FERM-P
"****" and "FERM-P ****" have been deposited at the Institute of Biotechnology, Institute of Industrial Science and Technology, and the mycological properties of these strains have been clarified by the aforementioned literature. Streptomyces ferreus JCM4031
(A strain having the same origin as the conserved strain NRRL2251 of "Agricultural Research Culture Collection";
U.S. Pat. No. 2,693,433) is a preserved strain of the microbial strain preservation facility of RIKEN, and is FERM-P *****
* Has been deposited at the Institute of Biotechnology, Institute of Industrial Science and Technology, and the mycological properties of this strain have been clarified by the above-mentioned literature.
【0008】また、放線菌の菌体または胞子を例えば紫
外線照射処理やNTG(N-methyl-N'-nitro-N-nitrosog
uanidine)処理等をすることによって変異させ、より好
ましい菌株を取得して用いることもできる。Further, the cells or spores of actinomycetes are treated with, for example, ultraviolet rays or NTG (N-methyl-N'-nitro-N-nitrosog).
It is also possible to mutate it by subjecting it to uanidine) treatment or the like to obtain and use a more preferable strain.
【0009】本発明における反応に必要な放線菌の菌体
を得るためには、好気条件下で菌株の培養を培地中で行
う。培地は主として液体培地を用い、炭素源としてグル
コース、マルトース、デキストロース、スターチ、オー
トミール、アラビノース、キシロース、シュクロースを
単独又は混合して用いる。窒素源としては、ペプトン、
カゼイン、ポリペプトン、大豆ペプトン、酵母エキス、
肉エキス、コーンスチープリカー、マルトエキス、綿実
粕、グルテンミール及び大豆粉などを単独又は混合して
用いる。その他、本菌株の生育を助け、12−ヒドロキ
シ−6−O−アルキルエリスロマイシン及びその類縁体
の生産を促進する有機物及び無機塩を必要により添加す
ることができる。培養方法は振盪培養、通気撹拌培養等
の好気的条件下の培養が適しており、pH6〜8、26
〜30℃で2〜7日間培養する。In order to obtain the cells of actinomycetes required for the reaction in the present invention, the strain is cultured in a medium under aerobic conditions. As the medium, a liquid medium is mainly used, and glucose, maltose, dextrose, starch, oatmeal, arabinose, xylose, and sucrose are used alone or as a mixture as a carbon source. As a nitrogen source, peptone,
Casein, polypeptone, soybean peptone, yeast extract,
Meat extract, corn steep liquor, malt extract, cottonseed meal, gluten meal, soybean flour and the like are used alone or in combination. In addition, organic substances and inorganic salts that help the growth of this strain and promote the production of 12-hydroxy-6- O- alkylerythromycin and its analogs can be added if necessary. As for the culture method, culture under aerobic conditions such as shaking culture and aeration stirring culture is suitable, and the pH is 6 to 8, 26.
Incubate at -30 ° C for 2-7 days.
【0010】この培養により得られた放線菌菌体または
その菌体が産生する酵素を含有する溶液を12−ヒドロ
キシ−6−O−アルキルエリスロマイシン及びその類縁
体を生産する反応に用いる。すなわち、培養中の菌体を
含む培養液に基質を添加するか、培養終了後遠心分離ま
たは濾過により分離した菌体を溶液に懸濁したものに基
質を添加して反応を開始する。培養終了後に遠心分離ま
たは濾過により分離した菌体を破砕処理してその産生す
る酵素を抽出しこれに基質を添加して反応することもで
きる。また、反応液中にα−シクロデキストリン、β−
シクロデキストリン、γ−シクロデキストリン等のシク
ロデキストリン類や、アンバーライトXAD−7[商品
名;オルガノ(株)製]等の吸着剤等の有機物、あるい
はツイーン80等の界面活性剤や塩化マグネシウムなど
の無機塩を、反応を促進させる目的で添加することがで
きる。さらにまた、放線菌菌体やその菌体が産生する酵
素は光架橋性樹脂プレポリマー、たとえばENT340
0[商品名;関西ペイント(株)製]などや、ウレタン
・プレポリマー、たとえばPU−9[商品名;東洋ゴム
(株)製]などや、κ−カラギナンなどの多糖類に固定
化してから溶液に添加してもよい。また、前期菌体の凍
結乾燥したものを上記と同様に用いることもできる。The actinomycete cells obtained by this culture or a solution containing the enzyme produced by the cells is used in the reaction for producing 12-hydroxy-6- O- alkylerythromycin and its analogs. That is, the substrate is added to the culture solution containing the microbial cells in culture, or the substrate is added to a suspension of the microbial cells separated by centrifugation or filtration after the completion of the culture to start the reaction. After the completion of the culture, the bacterial cells separated by centrifugation or filtration may be crushed to extract the enzyme produced, and a substrate may be added to this to react. In addition, α-cyclodextrin, β-
Cyclodextrins such as cyclodextrin and γ-cyclodextrin, organic substances such as adsorbents such as Amberlite XAD-7 [trade name; manufactured by Organo Corporation], surfactants such as Tween 80 and magnesium chloride Inorganic salts can be added for the purpose of promoting the reaction. Furthermore, actinomycetes and enzymes produced by the cells are photocrosslinkable resin prepolymers such as ENT340.
0 [trade name; manufactured by Kansai Paint Co., Ltd.], urethane prepolymers such as PU-9 [trade name; manufactured by Toyo Tire & Rubber Co., Ltd.], and after being immobilized on polysaccharides such as κ-carrageenan It may be added to the solution. Also, freeze-dried cells of the pre-stage cells can be used in the same manner as above.
【0011】本発明において用いられる反応溶液は、前
記培地を用いて培養した培養液であるか、あるいはトリ
ス酢酸、トリス塩酸、コハク酸ナトリウム−コハク酸、
コハク酸カリウム−コハク酸、クエン酸ナトリウム−ク
エン酸、カリウム及びナトリウムなどのリン酸塩、カコ
ジル酸ナトリウム−塩酸、イミダゾール−塩酸、ホウ酸
−ホウ砂などの緩衝液を単独または混合したものであ
る。その他溶液には目的の12−ヒドロキシ−6−O−
アルキルエリスロマイシン及びその類縁体の生産を促進
させる有機物、界面活性剤及び無機塩を必要により添加
することができる。The reaction solution used in the present invention is a culture solution obtained by culturing using the above-mentioned medium, or tris-acetic acid, tris-hydrochloric acid, sodium succinate-succinic acid,
Potassium succinate-succinic acid, sodium citrate-citric acid, phosphates such as potassium and sodium, buffer solutions such as sodium cacodylate-hydrochloric acid, imidazole-hydrochloric acid, boric acid-borax, etc., alone or mixed. . Other solutions include the desired 12-hydroxy-6- O-
If necessary, an organic substance, a surfactant and an inorganic salt that accelerate the production of alkylerythromycin and its analogs can be added.
【0012】本発明においては、前記菌体を含有する溶
液中で振盪操作、通気撹拌操作等に付して好気条件下で
行うことが適しており、pH5〜9、18〜37℃で1
〜10日間撹拌振盪する。また、酸素気流下で反応する
ことができる。基質は撹拌振盪開始時に適量添加する。
また、培養中の菌体を含む培養液を用いる場合は基質を
添加後、さらに上記と同条件下で反応を行う。これらの
反応により製造された12−ヒドロキシ−6−O−アル
キルエリスロマイシン及びその類縁体を単離するには培
養液からエリスロマイシンAなどのマクロライド抗生物
質を採取する一般的な方法に準じて行えばよい。例え
ば、反応終了後反応液を酢酸エチルまたは塩化メチレン
等の有機溶媒で抽出する。この有機溶媒層を分取し濃縮
した後、順相シリカゲルカラム(キーゼルゲル60F2
54,メルク社製)及び逆相シリカゲルカラム(ローバ
ーカラム,リクロプレップC18,メルク社製)を用い
て精製することにより目的の12−ヒドロキシ−6−O
−メチルエリスロマイシン及びその類縁体を単離するこ
とができる。また、セファデックスLH−20(ファル
マシア社製)等を用いた分配クロマトグラフィーやシリ
カゲルの薄層クロマトグラフィー及びダイヤイオンHP
−20(商品名;三菱化成工業社製)等によっても12
−ヒドロキシ−6−O−アルキルエリスロマイシン及び
その類縁体を単離することができる。In the present invention, it is suitable to carry out a shaking operation, an aeration stirring operation, etc. in a solution containing the above-mentioned bacterial cells under aerobic conditions, and the pH is 5 to 9 and 18 to 37 ° C.
Shake for 10 days. Further, the reaction can be performed under an oxygen stream. An appropriate amount of substrate is added at the start of stirring and shaking.
When using a culture solution containing cells in culture, the substrate is added, and then the reaction is performed under the same conditions as above. In order to isolate 12-hydroxy-6- O- alkylerythromycin and its analogs produced by these reactions, a general method for collecting macrolide antibiotics such as erythromycin A from a culture solution can be performed according to a general method. Good. For example, after completion of the reaction, the reaction solution is extracted with an organic solvent such as ethyl acetate or methylene chloride. After separating and concentrating the organic solvent layer, a normal phase silica gel column (Kieselgel 60F2
54, manufactured by Merck & Co., Inc. and a reversed-phase silica gel column (row bar column, lycroprep C18, manufactured by Merck & Co., Inc.) for purification of desired 12-hydroxy-6- O.
-Methylerythromycin and its analogs can be isolated. Also, partition chromatography using Sephadex LH-20 (manufactured by Pharmacia), thin layer chromatography on silica gel, and Diaion HP.
-20 (trade name; manufactured by Mitsubishi Kasei Co., Ltd.)
-Hydroxy-6- O- alkylerythromycin and its analogs can be isolated.
【0013】[0013]
【発明の効果】本発明の方法により、12位に水素原子
を有する6−O−アルキルエリスロマイシン及びその類
縁体の12位を直接水酸化し12−ヒドロキシ−6−O
−アルキルエリスロマイシン及びその類縁体を製造する
ことが可能になった。By the method of the present invention, 6- O having a hydrogen atom at position 12 - alkyl erythromycin and 12 of its analogs directly hydroxylating 12-hydroxy-6-O
It has become possible to produce alkyl erythromycin and its analogs.
【0014】[0014]
【実施例】以下、実施例を挙げて本発明を更に具体的に
説明する。 実施例1 スターチ2.4%、デキストロース0.1%、ペプトン
0.5%、肉エキス0.3%、酵母エキス0.5%、炭
酸カルシウム0.2%からなるpH7.0の無菌液体培
地(以下GM培地と略す)40mlの入った200ml
の枝付き三角フラスコにミクロモノスポア・メガロミセ
ア亜種ニグラ IFO14114株を1白金耳接種し、
28℃で96時間撹拌振盪培養した。培養終了後、培養
液を遠心分離して菌体を集め、この菌体を15mMトリ
ス塩酸、25mMコハク酸ナトリウム、2mM塩化マグ
ネシウムからなる、pH7.4の緩衝液(以下緩衝液A
と略す)40mlに懸濁し、再びこの懸濁液を遠心分離
して菌体を集めた。この菌体を緩衝液A 40mlに十
分撹拌して懸濁し、200mlの枝付き三角フラスコに
いれた。この三角フラスコに400μlのエタノールに
溶解した4mgの6−O−メチルエリスロマイシンB
[文献:ザ・ジャーナル・オブ・アンチバイオチクス
(The Journal of Antibiotics),第43巻,第5号,
第544ページ,1990年]を添加し、28℃で4日
間撹拌振盪して反応を行った。反応終了後の反応液のp
Hを炭酸水素ナトリウムで10に調整後、塩化メチレン
40mlで抽出した塩化メチレン層を減圧条件下で濃縮
した後、順相シリカゲルの薄層クロマトグラフィー(薄
層プレート:キーゼルゲル 1.05715,メルク社
製,展開溶媒:塩化メチレン:メタノール:25%アン
モニア水=100:10:1)に付した。展開後これを
ヨード発色させ、クラリスロマイシン標品(大正製薬
製,文献:米国特許第4331803号及び米国特許第
4990602号,以下実施例において同じ)と一致す
る部分のシリカゲルを掻き取り、酢酸エチルで抽出し
た。この酢酸エチル抽出物を減圧濃縮した後、逆相シリ
カゲルの薄層クロマトグラフィー(薄層プレート:キー
ゼルゲルArt.5747,メルク社製,展開溶媒;メ
タノール:水:酢酸アンモニウム15%溶液=5:2:
1)に付した。展開後これをヨード発色させ、クラリス
ロマイシン標品と一致するシリカゲル部分を掻き取り、
酢酸エチルで抽出した。この酢酸エチル抽出物を減圧濃
縮乾固し生成物1.2mgを得た。これはクラリスロマ
イシン標品と液体クロマトグラフィー(TSK−gel
ODS−120A:東ソー製,展開溶媒:アセトニト
リル:メタノール:50mM リン酸ナトリウム pH
6.8=6:2:3,流速1ml/min,カラム温度
50℃,フォトダイオードアレイ検出器(日本ミリポア
製)で検出)の保持時間、紫外線吸収スペクトル、マス
スペクトル開裂パターンが完全に一致した。 液体クロマトグラフィーの保持時間: 8.7分 UV: λmax =288nm(アセトニトリル) FAB−MS(m/z):748([M+H]+),5
90([M+H]+−cladinose),158。EXAMPLES The present invention will be described in more detail below with reference to examples. Example 1 Sterile liquid medium of pH 7.0 consisting of starch 2.4%, dextrose 0.1%, peptone 0.5%, meat extract 0.3%, yeast extract 0.5%, calcium carbonate 0.2%. (Hereinafter abbreviated as GM medium) 200 ml containing 40 ml
Inoculate an Erlenmeyer flask with branches with 1 platinum loop of Micromonospore megalomicea subsp. Nigra IFO14114 strain,
Culture was carried out at 28 ° C. for 96 hours with shaking. After the completion of the culture, the culture solution was centrifuged to collect the cells, and the cells were mixed with 15 mM Tris-hydrochloric acid, 25 mM sodium succinate, and 2 mM magnesium chloride at a pH of 7.4 (hereinafter referred to as buffer solution A).
It is suspended in 40 ml, and the suspension is centrifuged again to collect the cells. The cells were sufficiently stirred and suspended in 40 ml of buffer solution A, and placed in a 200 ml Erlenmeyer flask with a branch. In this Erlenmeyer flask, 4 mg of 6- O -methylerythromycin B dissolved in 400 μl of ethanol was added.
[Reference: The Journal of Antibiotics, Vol. 43, No. 5,
Page 544, 1990] was added, and the reaction was carried out by stirring and shaking at 28 ° C. for 4 days. P of the reaction solution after completion of the reaction
After adjusting H to 10 with sodium hydrogen carbonate, the methylene chloride layer extracted with 40 ml of methylene chloride was concentrated under reduced pressure, and then subjected to thin layer chromatography on normal phase silica gel (thin layer plate: Kieselgel 1.05715, manufactured by Merck & Co., Inc.). , Developing solvent: methylene chloride: methanol: 25% aqueous ammonia = 100: 10: 1). After development, this was subjected to iodine color development, and the silica gel in the portion corresponding to the clarithromycin standard product (manufactured by Taisho Pharmaceutical Co., Ltd., reference: US Pat. No. 4,331,803 and US Pat. No. 4,990,602, the same in the following examples) was scraped off, and ethyl acetate It was extracted with. The ethyl acetate extract was concentrated under reduced pressure, and then subjected to thin layer chromatography on reverse-phase silica gel (thin layer plate: Kieselgel Art.5747, manufactured by Merck & Co., a developing solvent; methanol: water: ammonium acetate 15% solution = 5: 2:
It is attached to 1). After the development, this is colored with iodine, and the silica gel part that matches the clarithromycin standard is scraped off.
It was extracted with ethyl acetate. The ethyl acetate extract was concentrated under reduced pressure to dryness to obtain 1.2 mg of a product. This is a clarithromycin standard and liquid chromatography (TSK-gel
ODS-120A: manufactured by Tosoh, developing solvent: acetonitrile: methanol: 50 mM sodium phosphate pH
6.8 = 6: 2: 3, flow rate 1 ml / min, column temperature 50 ° C., retention time of photodiode array detector (manufactured by Nippon Millipore)), ultraviolet absorption spectrum, mass spectrum cleavage pattern completely matched. . Retention time in liquid chromatography: 8.7 minutes UV: λ max = 288 nm (acetonitrile) FAB-MS (m / z): 748 ([M + H] + ), 5
90 ([M + H] + -cladinose), 158.
【0015】実施例2 滅菌したGM培地1Lの入った内容量1.8Lのジャー
3基のそれぞれにあらかじめGM培地で培養したミクロ
モノスポア・メガロミセア亜種ニグラ IFO1411
4株の培養液40mlを接種し、28℃で72時間通気
撹拌培養した。この培養液を遠心集菌し緩衝液A2Lに
懸濁後、再度遠心集菌した。この菌体を緩衝液A3Lに
懸濁した懸濁液を、先の培養で用いた内容量1.8Lの
ジャー3基のそれぞれに1Lずつ入れた。これに基質6
−O−メチルエリスロマイシンBをジャー1基あたり2
00mg添加し、28℃で通気撹拌し反応を96時間行
った。反応終了後の反応液を集め、そのpHを炭酸水素
ナトリウムで10に調整後、酢酸エチル1.5Lで抽出
した。これを減圧条件下で濃縮した後、順相シリカゲル
のカラムクロマトグラフィー(ボンドエルート:商品
名,ユニフレックス社製,サイズ20ml容,溶出溶
媒:塩化メチレン−メタノール−25%アンモニア水=
10:1:0.1)に付した。140mlの溶出溶媒を
濃縮し、赤褐色の油状濃縮物を得た。これを5℃で12
時間放置後、得られた粗結晶を5mlのアセトニトリル
−メタノール−塩化メチレン=10:2:1に溶解し、
逆相シリカゲルカラムクロマトグラフィー(ローバーカ
ラム・サイズB リクロプレップC18;商品名,メル
ク社製,25mmI.D.×310mm,溶出溶媒:ア
セトニトリル−メタノール−50mM リン酸ナトリウ
ム緩衝液 pH6.8=6:2:3)に付した。クラリ
スロマイシン溶出画分を逆相シリカゲルの薄層クロマト
グラフィー(薄層プレート:キーゼルゲルArt.57
47,メルク社製,展開溶媒;メタノール:水:酢酸ア
ンモニウム15%溶液=5:2:1)で確認し、その溶
出画分を集めて濃縮乾固した。これをエタノールにより
再結晶しクラリスロマイシン46.4mgを得た。これ
はクラリスロマイシン標品と融点、紫外線吸収スペクト
ル、マススペクトル開裂パターン、IR吸収スペクトル
及び13C−NMRが完全に一致した。Example 2 Micromonospore megalomicea subsp. Nigra IFO1411, which was previously cultivated in GM medium in each of three jars having a volume of 1.8 L containing 1 L of sterilized GM medium
40 ml of the culture solution of 4 strains was inoculated, and the cells were cultured at 28 ° C. for 72 hours with aeration and stirring. The culture solution was collected by centrifugation, suspended in buffer A2L, and then collected again by centrifugation. A suspension prepared by suspending the cells in buffer A3L was added to each of the three jars having the internal volume of 1.8 L used in the above culture, 1 L each. Substrate 6
- O - methylerythromycin B jar per group 2
00 mg was added, and the reaction was carried out at 28 ° C. with aeration and stirring for 96 hours. After completion of the reaction, the reaction solutions were collected, adjusted to pH 10 with sodium hydrogen carbonate, and extracted with 1.5 L of ethyl acetate. After concentrating this under reduced pressure, column chromatography on normal phase silica gel (Bond Elute: trade name, manufactured by Uniflex, size 20 ml, elution solvent: methylene chloride-methanol-25% aqueous ammonia =
10: 1: 0.1). 140 ml of the eluting solvent was concentrated to obtain a reddish brown oily concentrate. This is 12 at 5 ℃
After standing for a period of time, the obtained crude crystals were dissolved in 5 ml of acetonitrile-methanol-methylene chloride = 10: 2: 1,
Reversed-phase silica gel column chromatography (Rover column, size B, Licroprep C18; trade name, manufactured by Merck, 25 mm ID × 310 mm, elution solvent: acetonitrile-methanol-50 mM sodium phosphate buffer pH 6.8 = 6: 2: 3). The clarithromycin elution fraction was subjected to thin layer chromatography on reversed-phase silica gel (thin layer plate: Kieselgel Art.57.
47, manufactured by Merck & Co., Inc., a developing solvent; methanol: water: 15% ammonium acetate solution = 5: 2: 1), and the elution fractions were collected and concentrated to dryness. This was recrystallized from ethanol to obtain 46.4 mg of clarithromycin. This was in perfect agreement with the clarithromycin standard in melting point, ultraviolet absorption spectrum, mass spectrum cleavage pattern, IR absorption spectrum and 13 C-NMR.
【0016】m.p.: 225〜227℃ UV: λmax =288nm(アセトニトリル) FAB−MS(m/z):748([M+H]+),5
90([M+H]+−cladinose),158 IR(KBr)νmax =3474,1733,1693
cm-1 13 C−NMR(100.4MHz,CDCl3) δ
(ppm):221.0,175.8,102.7,9
6.1,80.9,78.4,78.3,77.9,7
6.6,74.2,72.7,70.9,69.0,6
8.6,65.8,65.6,50.6,49.5,4
5.2,45.0,40.3,39.3,39.1,3
7.2,34.9,29.0,21.5,21.4,2
1.0,19.7,18.7,18.0,15.9,1
2.3,10.6,9.1。M. p. : 225-227 degreeC UV: (lambda) max = 288nm (acetonitrile) FAB-MS (m / z): 748 ([M + H] < +>), 5.
90 ([M + H] + -cladinose), 158 IR (KBr) ν max = 3474,1733,1693
cm -1 13 C-NMR (100.4 MHz, CDCl 3 ) δ
(Ppm): 221.0, 175.8, 102.7, 9
6.1, 80.9, 78.4, 78.3, 77.9, 7
6.6, 74.2, 72.7, 70.9, 69.0, 6
8.6, 65.8, 65.6, 50.6, 49.5, 4
5.2, 45.0, 40.3, 39.3, 39.1, 3
7.2, 34.9, 29.0, 21.5, 21.4, 2
1.0, 19.7, 18.7, 18.0, 15.9, 1
2.3, 10.6, 9.1.
【0017】実施例3 実施例1における操作で使用菌株をミクロモノスポア・
メガロミセア亜種ニグラ IFO14114株の代わり
に、サッカロポリスポラ・エリスラエア IFO134
26株及びストレプトミセス・フェレウス JCM40
31を用いて実施した。なお、サッカロポリスポラ属及
びストレプトミセス属の菌株では、オートミール2.0
%、グルコース2.0%、肉エキス0.3%、塩化ナト
リウム0.3%、硫酸第2鉄0.04%、塩化マンガン
0.04%、炭酸カルシウム0.3%、pH7.0から
なる無菌液体培地を培養に用いた。その結果、各菌株に
クラリスロマイシンの生成が認められた。各菌株におけ
るクラリスロマイシン生産量を以下の表1に記す。Example 3 The strain used in the procedure of Example 1 was micromonospores.
Saccharopolispora erythraea IFO134 in place of the megalomicea subsp. Nigra IFO14114 strain
26 strains and Streptomyces ferreus JCM40
31 was used. For strains of the genera Saccharopolispora and Streptomyces, oatmeal 2.0
%, Glucose 2.0%, meat extract 0.3%, sodium chloride 0.3%, ferric sulfate 0.04%, manganese chloride 0.04%, calcium carbonate 0.3%, pH 7.0 Sterile liquid medium was used for the culture. As a result, the production of clarithromycin was confirmed in each strain. The clarithromycin production in each strain is shown in Table 1 below.
【0018】[0018]
【表1】 [Table 1]
【0019】実施例4 実施例1における操作で、使用菌株にサッカロポリスポ
ラ・エリスレア IFO13426株を用い、基質6−
O−メチルエリスロマイシンBを添加すると同時に終濃
度0.2%になるようにβ−シクロデキストリン(メル
シャン社製)を添加して実施した結果、クラリスロマイ
シン0.4mgを得た。 実施例5 実施例2において反応時間を48時間にして実施した結
果、191mgのクラリスロマイシンを得た。Example 4 In the procedure of Example 1, Saccharopolyspora erythrea IFO 13426 was used as the strain to be used, and the substrate 6-
When O -methylerythromycin B was added and β-cyclodextrin (manufactured by Mercian) was added at a final concentration of 0.2%, the result was 0.4 mg of clarithromycin. Example 5 As a result of carrying out the reaction in Example 2 with a reaction time of 48 hours, 191 mg of clarithromycin was obtained.
【0020】実施例6 ミクロモノスポア・メガロミセア亜種ニグラ IFO1
4114株及びサッカロポリスポラ・エリスレア IF
O13426株をそれぞれGM培地及びJ3培地(グル
コース2%、バクトソイトン(商品名:デフィコ社製)
1.5%、コーンスチープリカー0.5%、食塩0.4
%、リン酸水素2カリウム0.04%、塩化マンガン4
水和物0.04%、酵母エキス0.2%、炭酸カルシウ
ム0.2%,pH7.0)各80mlを用いて28℃、
3日間通気撹拌培養した後、緩衝液C(トリス−塩酸8
0mM、pH7.5、ジチオスレイトール2mM、グリ
セロール15%)で遠心洗浄した。得られたミクロモノ
スポア・メガロミセア亜種ニグラ IFO14114株
及びサッカロポリスポラ・エリスレア IFO1342
6株の洗浄菌体を緩衝液Cに懸濁後、超音波処理して破
砕し、菌体を遠心除去後、上清画分として各40mlの
粗酵素液を得た。酵素反応は得られた各粗酵素液の一部
にNADPH0.26mM、フェレドキシンNADPリ
ダクターゼ0.4ユニット、フェレドキシン1.6m
g、グルコース6リン酸14mM、グルコース6リン酸
デヒドロゲナーゼ1ユニット、ニコチンアミド10m
M、6−O−メチルエリスロマイシンB0.4mgを添
加した反応液2ml中で30℃、12時間通気撹拌する
ことにより行った。各反応液をそれぞれ酢酸エチルで抽
出し得られたクラリスロマイシンの生成量を液体クロマ
トグラフィー(TSK−gel ODS−120A:東
ソー製,展開溶媒:アセトニトリル:メタノール:50
mM リン酸ナトリウム pH6.8=6:2:3,流
速1ml/min,カラム温度50℃,フォトダイオー
ドアレイ検出器(日本ミリポア製)で検出,定量)で分
析した。結果を以下の表2に記す。Example 6 Micromonospore megalomicea subsp. Nigra IFO1
4114 strain and Saccharopolyspora erythrea IF
The O13426 strain was used as a GM medium and a J3 medium (glucose 2%, Bactosoyton (trade name: manufactured by Defico), respectively.
1.5%, corn steep liquor 0.5%, salt 0.4
%, Dibasic hydrogen phosphate 0.04%, manganese chloride 4
Hydrate 0.04%, yeast extract 0.2%, calcium carbonate 0.2%, pH 7.0) 28 ° C. using 80 ml of each
After culturing with aeration and stirring for 3 days, buffer solution C (Tris-hydrochloric acid 8
It was washed by centrifugation with 0 mM, pH 7.5, dithiothreitol 2 mM, glycerol 15%). Obtained Micromonospore megalomicea subsp. Nigra IFO14114 strain and Saccharopolyspora erythrea IFO1342
The washed bacterial cells of the 6 strains were suspended in buffer C, sonicated and disrupted, and the bacterial cells were removed by centrifugation to obtain 40 ml of a crude enzyme solution as a supernatant fraction. In the enzyme reaction, NADPH 0.26 mM, ferredoxin NADP reductase 0.4 unit, ferredoxin 1.6 m was added to a part of each crude enzyme solution obtained.
g, glucose 6-phosphate 14 mM, glucose 6-phosphate dehydrogenase 1 unit, nicotinamide 10 m
It was carried out by aeration stirring at 30 ° C. for 12 hours in 2 ml of a reaction solution containing 0.4 mg of M, 6- O -methylerythromycin B. The amount of clarithromycin produced by extracting each reaction solution with ethyl acetate was measured by liquid chromatography (TSK-gel ODS-120A: manufactured by Tosoh Corporation, developing solvent: acetonitrile: methanol: 50).
mM sodium phosphate pH 6.8 = 6: 2: 3, flow rate 1 ml / min, column temperature 50 ° C., detection with photodiode array detector (manufactured by Nippon Millipore), quantitative analysis). The results are shown in Table 2 below.
【0021】[0021]
【表2】 [Table 2]
【0022】実施例7 6,4”−ジ−O−メチルエリスロマイシンAの製造:
文献[ザ・ジャーナル・オブ・アンチバイオチクス(Th
e Journal of Antibiotics),第43巻,第5号,第5
44頁,1990年]に記載されている6,4”−ジ−
O−メチルエリスロマイシンB 4.0mgを原料に用
いて実施例1と同様の方法に従って6,4”−ジ−O−
メチルエリスロマイシンA(融点:208〜210℃,
13C−NMR:図1)を1.0mg得た。Example 7 Preparation of 6,4 "-di- O -methylerythromycin A:
Reference [The Journal of Antibiotics (Th
e Journal of Antibiotics), Vol. 43, No. 5, No. 5
P. 44, 1990] 6,4 "-Ji-
According to the same method as in Example 1 using 4.0 mg of O -methylerythromycin B as a raw material, 6,4 ″ -di- O—
Methylerythromycin A (melting point: 208-210 ° C,
13 C-NMR: FIG. 1) (1.0 mg) was obtained.
【0023】実施例8 6,11,4”−トリ−O−メチルエリスロマイシンA
の製造:実施例7で挙げた文献に記載の6,11,4”
−トリ−O−メチルエリスロマイシンB 4.0mgを
原料に用いて実施例1と同様の方法に従って6,11,
4”−トリ−O−メチルエリスロマイシンA[文献:ザ
・ジャーナル・オブ・アンチバイオチクス(The Journa
l of Antibiotics),第43巻,第3号,第286頁,
1990年]を1.0mg得た。Example 8 6,11,4 "-tri- O -methylerythromycin A
Production: 6,11,4 "described in the literature cited in Example 7
-Tri- O -methylerythromycin B 4.0 mg was used as a starting material according to the same method as in Example 1,6,11,
4 "-tri- O -methylerythromycin A [Reference: The Journal of Antibiotics]
l of Antibiotics), Vol. 43, No. 3, p. 286,
1990].
【0024】実施例9 デクラジノシル−6−O−メチルエリスロマイシンAの
製造:6−O−メチルエリスロマイシンBを酸処理[文
献:エクスペリエンチア(Experientia )第27巻,第
362頁,1971年]することによって得られたデク
ラジノシル−6−O−メチルエリスロマイシンB(融
点:162〜164℃,13C−NMR:図2)を合成し
その4.0mgを原料に用いて実施例1と同様の方法に
従ってデクラジノシル−6−O−メチルエリスロマイシ
ンA[文献:ケモテラピー(Chemotherapy),第36巻
(S−3),第264ページ,1988年]0.6mg
を得た。Example 9 Preparation of decladinosyl-6- O -methylerythromycin A: 6- O -methylerythromycin B is acid-treated [Reference: Experientia Vol. 27, p. 362, 1971]. The decladinosyl-6- O -methylerythromycin B (melting point: 162-164 ° C, 13 C-NMR: Fig. 2) thus obtained was synthesized, and 4.0 mg thereof was used as a starting material according to the same method as in Example 1. Decladinosyl-6- O -methylerythromycin A [Reference: Chemotherapy, Volume 36 (S-3), 264, 1988] 0.6 mg
I got
【0025】実施例10 (14R)−14−ヒドロキシ−6−O−メチルエリス
ロマイシンAの製造 (14R)−14−ヒドロキシ−6−O−メチルエリス
ロマイシンB[文献:ザ・ジャーナル・オブ・アンチバ
イオチクス(The Journal of Antibiotics)第41巻,
第7号,908頁〜915頁,1988年]4.0mg
を原料に用いて実施例1と同様の方法に従って(14
R)−14−ヒドロキシ−6−O−メチルエリスロマイ
シンA(文献:上記と同じ)1.0mgを得た。[0025] Example 10 (14 R) -14- hydroxy-6-O - producing (14 R) methyl erythromycin A-14-hydroxy-6-O - methylerythromycin B [Reference: The Journal of Anti Biotics (The Journal of Antibiotics) Vol. 41,
No. 7, pp. 908-915, 1988] 4.0 mg.
In the same manner as in Example 1 using
R ) -14-Hydroxy-6- O -methylerythromycin A (literature: same as above) (1.0 mg) was obtained.
【図1】重クロロホルム中、75MHzで測定した6,
4”−ジ−O−メチルエリスロマイシンAの13C−NM
Rスペクトルを示す。FIG. 1: 6, measured in deuterated chloroform at 75 MHz
13 C-NM of 4 "-di- O -methylerythromycin A
The R spectrum is shown.
【図2】重クロロホルム中、75MHzで測定したデク
ラジノシル−6−O−メチルエリスロマイシンBの13C
−NMRスペクトルを示す。FIG. 2: 13 C of decladinosyl-6- O -methylerythromycin B measured at 75 MHz in deuterated chloroform.
-Shows an NMR spectrum.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 森本 繁夫 東京都豊島区高田3丁目24番1号 大正製 薬株式会社内 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Shigeo Morimoto 3-24-1 Takada, Toshima-ku, Tokyo Taisho Pharmaceutical Co., Ltd.
Claims (3)
その類縁体を水酸化する放線菌菌体またはその菌体が産
生する酵素を含有する溶液中に、12位に水素原子を有
する6−O−アルキルエリスロマイシン及びその類縁体
を加えてその水素原子を水酸基に変換することを特徴と
する12−ヒドロキシ−6−O−アルキルエリスロマイ
シン及びその類縁体の製造方法。1. A 6- O -alkyl having a hydrogen atom at the 12-position in a solution containing a 6- O- alkylerythromycin and an actinomycete that hydroxylates actinomycetes or an enzyme produced by the fungus. A method for producing a 12-hydroxy-6- O -alkylerythromycin and its analogs, which comprises adding erythromycin and its analogs to convert the hydrogen atom into a hydroxyl group.
キルエリスロマイシン及びその類縁体が、下記式で表さ
れる請求項1に記載の製造方法。 【化1】 [式中、R1は炭素原子数1〜4個のアルキル基を示
し、R2は水素原子またはメチル基を示し、R3は水素原
子または水酸基を示し、R4は水素原子または式 【化2】 (式中、R5は水素原子またはメチル基を示す。)で表
される基を示す。]2. The production method according to claim 1, wherein the 6- O- alkylerythromycin having a hydrogen atom at the 12-position and its analogs are represented by the following formula. Embedded image [In the formula, R 1 represents an alkyl group having 1 to 4 carbon atoms, R 2 represents a hydrogen atom or a methyl group, R 3 represents a hydrogen atom or a hydroxyl group, R 4 represents a hydrogen atom or a formula 2] (In the formula, R 5 represents a hydrogen atom or a methyl group). ]
その類縁体を水酸化する放線菌がミクロモノスポラ(Mi
cromonospora)属,ストレプトミセス(Streptomyces)
属,サッカロポリスポラ(Saccharopolyspora )属に属
する菌である請求項1または2に記載の製造方法。3. An actinomycete which hydroxylates 6- O- alkylerythromycin and its analogs is micromonospora ( Mi
cromonospora) genus Streptomyces (Streptomyces)
The method according to claim 1 or 2, which is a bacterium belonging to the genus Saccharopolyspora .
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP6232558A JPH0889277A (en) | 1994-09-28 | 1994-09-28 | Production of 12-hydroxy-6-o-alkylerythromycin and its homologue |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP6232558A JPH0889277A (en) | 1994-09-28 | 1994-09-28 | Production of 12-hydroxy-6-o-alkylerythromycin and its homologue |
Publications (1)
Publication Number | Publication Date |
---|---|
JPH0889277A true JPH0889277A (en) | 1996-04-09 |
Family
ID=16941216
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP6232558A Pending JPH0889277A (en) | 1994-09-28 | 1994-09-28 | Production of 12-hydroxy-6-o-alkylerythromycin and its homologue |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPH0889277A (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2003504071A (en) * | 1999-07-12 | 2003-02-04 | ジョジュシェルクタトー インテーゼト カーエフテー | Hydroxylation of compactin to pravastatin by micromonospora |
-
1994
- 1994-09-28 JP JP6232558A patent/JPH0889277A/en active Pending
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2003504071A (en) * | 1999-07-12 | 2003-02-04 | ジョジュシェルクタトー インテーゼト カーエフテー | Hydroxylation of compactin to pravastatin by micromonospora |
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