JP4210744B2 - Sphingolipid derivatives - Google Patents

Description

（式中、Ｒ ^１ はアルキレン基を、Ｒ ^２ とＲ ^３ はアルケニル基を、Ｒ ^４ とＲ ^５ はホスホコリン残基を、Ｒ ^６ とＲ ^７ は水素原子またはアルキル基をそれぞれ示す。）
【０００７】
【発明の実施の形態】
本発明の目的生成物である新規なスフィンゴ脂質誘導体は、下記一般式（１）で表すことができる。
一般式（１）
【化３】

（式中の各記号は前記と同じ。）
【０００８】
前記Ｒ ^１ はアルキレン基を示す。アルキレン基としては、その主鎖を構成する炭素数が、１〜６０、好ましくは１〜２５のアルキレン基が挙げられる。このようなアルキレン基の具体例としては、後記するアルキル基から水素原子を１個除いたものが例示される。
また前記Ｒ ^２ とＲ ^３ はアルケニル基を示す。アルケニル基としては、その主鎖を構成する炭素数が、２〜５０、好ましくは２〜２５のアルケニル基が挙げられる。このようなアルケニル基の具体例としては、たとえば、ビニル、プロペニル、ブテニル、アクリル、メタクリル、オクチニル、ドデセニル、ウンデセニルなどが例示される。
また、前記Ｒ ^４ とＲ ^５ はホスホコリン残基を示す。
更に前記Ｒ ^６ とＲ ^７ は、水素原子またはアルキル基を示す。アルキル基としては、その主鎖を構成する炭素数が１〜６０、好ましくは１〜２５のものが挙げられる。このようなアルキル基としては、たとえば、メチル、エチル、 n −プロピル、イソプロピル、 n −ブチル、イソブチル、ｓ e ｃ−ブチル、 t −ブチル、 n −ペンチル、イソペンチル、 2 −メチルブチル、 1 −メチルブチル、 n −へキシル、イソヘキシル、 3 −メチルペンチル、 2 −メチルペンチル、 1 −メチルペンチル、ヘプチル、オクチル、イソオクチル、 2 −エチルヘキシル、ノニル、デシル、ウンデシル、ドデシル、テトラデシル、ヘキサデシル、オクタデシル、エイコシルなどが例示される。
【００２７】
本発明の一般式（１）で示されるスフィンゴ脂質誘導体は、たとえば、次のようにして合成することができる。
まず、表１に示されるように、前記非特許文献２に記載の方法に準じてセリンもしくはセリン誘導体を出発原料とし、中間体であるガーナ−アルデヒドまで合成する。つぎにこのガーナ−アルデヒドにＲ^２もしくはＲ^３をカップリングさせ、脱アセトナイド、脱Ｂｏｃを行い、スフィンゴシン誘導体に導く。引き続き、Ｒ^４もしくはＲ^５を１級水酸基と反応させた後、Ｒ^６もしくはＲ^７を２級水酸基と反応させ、化合物ＡおよびＢを合成する。ついで両者をＲ^１とカップリング反応させることにより所望とするスフィンゴ脂質誘導体を得ることができる。また、後記表２に示すようにスフィンゴシン誘導体とＲ^１を反応させた後、Ｒ^４もしくはＲ^５を１級水酸基と反応させ、引き続きＲ^６もしくはＲ^７を２級水酸基と反応させることにより所望のスフィンゴ脂質誘導体を得ることもできる。
【００２８】
【表１】

【００２９】
本発明の一般式（１）で示されるスフィンゴ脂質誘導体は、同一分子内にセラミド誘導体を２分子有するので、セラミド１分子からなるスフィンゴ脂質誘導体は、細胞膜の外膜や内膜に集積するが、内膜と外膜の位置は異なる。同一分子内にセラミド誘導体を２分子有するスフィンゴ脂質誘導体は、細胞膜での集積時、外膜と内膜とを貫通した状態で存在すると考えられ、膜貫通型の界面活性としての性質を有することから、抗腫瘍剤、抗インフルエンザ剤等の医薬、保湿剤、育毛剤等の化粧品、農薬、工業薬品などに幅広く利用することができる。
【００３０】
【実施例】
以下、本発明につき実施例を挙げて説明するが、その要旨を越えない限り以下に限定されるものではない。
【００３１】
表２に示す合成スキームにしたがって、本発明のスフィンゴ脂質誘導体（化合物１２）を合成した。
【００３２】
［化合物１の合成］
化合物１は下記文献を参考に合成した。
Ｈｅｌｖ．Ｃｈｉｍ．Ａｃｔａ．，１９８８，３５４、Ｊ．Ｍｅｄ．Ｃｈｅｍ．，１９９９，２６８７、Ｂｉｏｏｒｇ．Ｍｅｄ．Ｃｈｅｍ．Ｌｅｔｔ．，１９９８，１７９
【００３３】
［化合物２の合成］
窒素雰囲気下、１−ヘプチン（３．８７ｍＬ，２９．４ｍｍｏｌ）を脱水テトラヒドロフランに溶解し−４０度で攪拌した。この溶液に、ｎ−ブチルリチウムのヘキサン溶液（１ｇ／ｍＬ，１８．９ｍＬ）を滴下し、同温度で１５分間攪拌させた。この反応溶液に、化合物１（２．２５ｇ，９．８２ｍｍｏｌ）のテトラヒドロフラン溶液を滴下し、−２０度まで１時間かけてゆっくりと加温した。この溶液に塩化アンモニウム飽和水溶液を加えた後、酢酸エチルで３回抽出操作を行った。有機層を飽和食塩水で洗浄し、硫酸ナトリウムで乾燥した。有機層を減圧留去して得られたシロップ状の粗生成物（３．２１５ｇ）を、シリカゲルカラムクロマトグラフィー（n-ヘキサン：酢酸エチル＝１０：１）で精製し、化合物２（２．３ｇ,、７２．１％）を得た。
【００３４】
［化合物３の合成］
化合物２（1．5ｇ，４．６１ｍｍｏｌ）をメタノールに溶かし、p-トルエンスルホン酸１水和物（１０ｍｇ）を加えて窒素雰囲気下室温で攪拌した。２５時間後、p-トルエンスルホン酸１水和物（２０ｍｇ）を加えてさらに２４時間攪拌した。メタノールを減圧留去して得られた残渣に、炭酸水素ナトリウム飽和水溶液と酢酸エチルを加えて分液した。水層をさらに酢酸エチルで２回抽出し、得られた有機層を飽和食塩水で洗浄し、硫酸ナトリウムで乾燥した。有機層を減圧留去して得られたシロップ状の粗生成物（１．３０５ｇ）を、シリカゲルカラムクロマトグラフィー（n-ヘキサン：酢酸エチル＝３：１から１：１）で精製し、化合物３（１．２５ｇ，９４．９％）を得た。
【００３５】
［化合物４の合成］
窒素雰囲気下、脱アセタール体（７２６ｍｇ，２．５５ｍｍｏｌ）を脱水エチルエーテルに溶かし、−２０度で攪拌させた。この溶液に、Ｒｅｄ−Ａｌのトルエン溶液（濃度６５％，３．８２ｍＬ，１２．７ｍｍｏｌ）を脱水エチルエーテルで希釈した溶液を滴下し、−２０度から室温まで１時間かけてゆっくりと加温した。この反応溶液を、さらに室温で２４時間攪拌させたのち、メタノール、飽和のロッシェル塩水溶液（５０ｍＬ）を加え、酢酸エチルで３回抽出した。得られた有機層を飽和食塩水で洗浄し、硫酸マグネシウムで乾燥した。有機層を減圧留去して得られた粗生成物を、シリカゲルカラムクロマトグラフィー（n-ヘキサン：酢酸エチル＝２：１から１：２）で精製し、化合物５（４８８ｍｇ，６６．７％）を得た。
【００３６】
［化合物５の合成］
化合物４（４８５ｍｇ，１．６９ｍｍｏｌ）をトリフルオロ酢酸（１．０ｍＬ）に溶かし、氷冷下１５分間攪拌後、室温にて３０分攪拌した。さらに、トリフルオロ酢酸（０．５ｍＬ）を加え、室温にて１時間攪拌した。トリフルオロ酢酸を減圧留去させ、２Mの水酸化ナトリウム水溶液（５ｍＬ）を加えエチルエーテルで５回抽出した。得られた有機層を硫酸ナトリウムで乾燥し、エチルエーテルを減圧留去して化合物５（３１０ｍｇ，９８％）を得た。
【００３７】
［化合物６の合成］
化合物５（１９ｍｇ、０．１ｍｍｏｌ）の脱水テトラヒドロフラン溶液に、調製したヘキサデカン二酸スクシンイミドエステル（１５ｍｇ、０．０５ｍｍｏｌ）を加え、窒素雰囲気下室温で攪拌した。この溶液に脱水トリエチルアミン（０．０１４ｍＬ、０．１ｍｍｏｌ）を加え、窒素雰囲気下室温で２４時間攪拌した。その後、溶媒を減圧留去し、得られた残渣をシリカゲルカラムクロマトグラフィー（n-ヘキサン：酢酸エチル：メタノール＝１０：１０：１）で精製し、化合物６（２３．１ｍｇ、７３．９％）を得た。
【００３８】
生成物は、^１Ｈ−ＮＭＲスペクトルにより同定した。
【００３９】
^１Ｈ−ＮＭＲ（ＴＭＳ、ＣＤＣｌ_３）：６．２８（２Ｈ、ｄ、Ｊ＝７．７Ｈｚ）、５．７９（２Ｈ、ｄｔｄ、Ｊ＝１５．３、６．７、1．０Ｈｚ）、５．５３（２Ｈ、ｄｄｔ、Ｊ＝１５．３、６．４、１．０Ｈｚ）、４．３２（２Ｈ、ｂｒｓ）、３．９５（２Ｈ、ｂｒｄ、Ｊ＝１１．９Ｈｚ）、３．９１（２Ｈ、ｍ）、３．７１（２Ｈ、ｍ）、２．８５（４Ｈ、ｂｒｓ）、２．２３（４Ｈ、ｔ、Ｊ＝７．４Ｈｚ）、２．０６（４Ｈ、ｄｔ、Ｊ＝７．５、６．７Ｈｚ）、１．６４（４Ｈ、ｔｔ、Ｊ＝７．４、７．４Ｈｚ）、１．３８（４Ｈ、ｔｔ、Ｊ＝７．４、７．４Ｈｚ）、１．２５−１．３０（１４Ｈ、ｍ）、０．８９（６Ｈ、ｔ、Ｊ＝７．０Ｈｚ）。
【００４０】
［化合物７の合成］
化合物６（１０ｍｇ、０．０１６ｍｍｏｌ）の無水ピリジン溶液に、Ｎ，Ｎ−ジメチルアミノピリジン（０．０３２ｍｇ、０．００２６ｍｍｏｌ）を加え、室温で２時間攪拌後、塩化ベンゾイル（０．００６ｍｌ、０．０５２ｍｍｏｌ）を加え、同温度で２０時間攪拌した。飽和塩化アンモニウム水溶液を加え激しく攪拌した後、クロロホルムで３回抽出した。得られた有機層を飽和塩化ナトリウム水溶液で洗浄し、硫酸ナトリウムで脱水した。有機層を減圧留去して得られた粗生成物をシリカゲルカラムクロマトグラフィー（n-ヘキサン：酢酸エチル＝２：１からn-ヘキサン：酢酸エチル：メタノール＝６：６：１）で精製し、化合物７（７．３ｍｇ、５４．８％）を得た。
【００４１】
生成物は、^１Ｈ−ＮＭＲスペクトルにより同定した。
【００４２】
^１Ｈ−ＮＭＲ（ＴＭＳ、ＣＤＣｌ_３）：８．０２（４Ｈ、ｄ、Ｊ＝８．３Ｈｚ）、７．５８（２Ｈ、ｔ、Ｊ＝７．６Ｈｚ）、７．４５（４Ｈ、ｔ、Ｊ＝８．０Ｈｚ）、５．９８（２Ｈ、ｄ、Ｊ＝７．９ Ｈｚ）、５．７７（２Ｈ、ｄｔ、Ｊ＝１５．５、６．７Ｈｚ）、５．５３（２Ｈ、ｄｄｔ、Ｊ＝１５．５、６．７、１．０Ｈｚ）、４．５６（２Ｈ、ｄｄ、Ｊ＝１２．５、８．３ Ｈｚ）、４．４１（４Ｈ、ｍ）、４．２７（２Ｈ、ｂｒｓ）、２．８７（２Ｈ、ｄ、Ｊ＝５．０Ｈｚ）、２．１９（４Ｈ、ｔ、Ｊ＝７．６Ｈｚ）、２．０１（４Ｈ、ｍ）、１．５８（４Ｈ、ｍ）、１．３４（４Ｈ、ｍ）、１．３４−１．２０（２８Ｈ、ｍ）、０．８７（６Ｈ、ｔ、Ｊ＝７．０Ｈｚ）。
【００４３】
［化合物８の合成］
氷冷下、化合物７（２０ｍｇ、０．０２４ｍｍｏｌ）の脱水ジクロロメタン溶液に２，６−ルチジン（０．０１１ｍｌ、０．０９６ｍｍｏｌ）を加え、同温度で１時間攪拌した。この溶液にｔ−ブチルジメチルシラントリフラート（０．０７２ｍｍｏｌ）をゆっくり滴下後、室温までゆっくり上昇させ、さらに室温で１時間攪拌した。飽和食塩水を加え攪拌し、ジクロロメタンで３回抽出を行った。得られた有機層を硫酸ナトリウムで乾燥し、減圧留去して化合物８を得た。
【００４４】
［化合物９の合成］
化合物８（２０ｍｇ、０．０２４ｍｍｏｌ）の脱水ジクロロメタン：脱水メタノール溶液（１：１）に炭酸カリウム（２０ｍｇ）を加え、冷蔵庫で３日間静置させ、化合物９を定量的に得た。
【００４５】
［化合物１０の合成］
５当量の２−ブロモエチルジクロロホスフェートの無水ジエチルエーテル溶液に、氷冷下、当量のトリエチルアミン、化合物９の無水ジエチルエーテル溶液を滴下した。同温にて３．５時間撹拌した後、０．５Ｎ塩化カリウム水溶液を加え、ジエチルエーテルで希釈し１時間撹拌した。ジエチルエーテルで抽出した後、水、飽和食塩水の順で洗浄した。分離した有機層を無水硫酸マグネシウムで乾燥、濾過した後、溶媒を減圧下にて留去し、化合物１０を得た。
【００４６】
[化合物１１の合成]
化合物１０をクロロホルム：イソプロピルアルコール：アセトニトリル（３：３：５）に溶解させ、３０％トリメチルアミン水溶液を加えて、５０℃にて４時間撹拌した。冷後、減圧下にて濃縮した。残渣をシリカゲルカラムクロマトグラフィー（クロロホルム：メタノール＝１０：１）で精製し、化合物１１を７４％で得た。
【００４７】
[化合物１２の合成]
化合物１１の無水テトラヒドロフラン溶液に氷冷下、２当量のテトラn-ブチルアンモニウムフルオリド（１．０Ｍテトラヒドロフラン溶液）を加え、室温にて４時間撹拌した。反応溶液を減圧下にて濃縮し、得られた残渣をシリカゲルカラムクロマトグラフィー（クロロホルム：メタノール＝５：１）で精製し、化合物１２を収率８８％で得た。
【００４８】
生成物は^１Ｈ−ＮＭＲスペクトルにより同定した。
【００４９】
^１Ｈ−ＮＭＲ（ＣＤＣｌ_３−ＣＤ_３ＯＤ、ＴＭＳ）：７．１９（２Ｈ、ｄ、Ｊ＝８．３Ｈｚ）、５．６４（２Ｈ、ｄｔｄ、Ｊ＝１５．３、６．７、1．０Ｈｚ）、５．４０（２Ｈ、ｄｄｔ、Ｊ＝１５．３、６．４、１．０Ｈｚ）、４．３２（２Ｈ、ｂｒｓ）、３．９２−３．８１（８Ｈ、ｍ）、３．７１（２Ｈ、ｍ）、３．４３（４Ｈ、ｍ）、３．２０（１８Ｈ、ｓ）、２．８５（２Ｈ、ｂｒｓ）、２．２３（４Ｈ、ｔ、Ｊ＝７．４Ｈｚ）、２．０６（４Ｈ、ｄｔ、Ｊ＝７．５、６．７Ｈｚ）、１．６４（４Ｈ、ｔｔ、Ｊ＝７．４、７．４Ｈｚ）、１．３８（４Ｈ、ｔｔ、Ｊ＝７．４、７．４Ｈｚ）、１．２５−１．３０（２８Ｈ、ｍ）、０．８９（６Ｈ、ｔ、Ｊ＝７．０Ｈｚ）。
【００５０】
【表２】

【００５１】
【発明の効果】
本発明によれば、同一分子内にセラミド誘導体を２分子有する新規スフィンゴ脂質誘導体が提供される。
本発明方法で得られるスフィンゴ脂質誘導体は、抗腫瘍剤、抗インフルエンザ剤等の医薬、保湿剤、育毛剤等の化粧品、農薬、工業薬品などに幅広く用いられる。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a novel sphingolipid derivative applicable to a wide range of uses such as pharmaceuticals such as antitumor agents and anti-influenza agents, cosmetics such as humectants and hair restorers, agricultural chemicals and industrial chemicals.
[0002]
[Prior art]
Conventionally, sphingolipid derivatives are known as important compounds widely used in medicines such as antitumor agents and anti-influenza agents, cosmetics such as moisturizers and hair restorers, agricultural chemicals and industrial chemicals.
As an artificial method for synthesizing such sphingolipid derivatives, development of a method for synthesizing sphingomyelin using serine or a serine derivative as a starting material has been mainly carried out. In particular, garner aldehyde, sphingosine, and ceramide, which are intermediates of sphingomyelin, have been developed. A synthetic route has been established for non-patent literature 1 (Non-Patent Document 1), and a total paper synthesis method for sphingomyelin is reported in a report by Slotte et al.
[0003]
However, sphingolipid-related compounds are in the midst of research despite their interest in properties and physical properties, and there are still many undocumented compounds. For example, ceramide derivatives in the same molecule No sphingolipid derivative having a molecule has been known.
[0004]
[Non-Patent Document 1]
Chem. Phys. Lipids, 1999, p. 3 [Non-Patent Document 2]
Biochemistry, 1991, pp. 10746 [0005]
[Problems to be solved by the invention]
An object of the present invention is to provide a novel sphingolipid derivative having two ceramide derivatives in the same molecule, which is useful as a medicine, cosmetics and the like.
[0006]
[Means for Solving the Problems]
As a result of intensive studies to solve the above problems, the present inventors have completed the present invention.
That is, according to the present invention, a novel sphingolipid derivative having two ceramide derivatives in the same molecule represented by the following general formula (1) is provided.
General formula (1)
[Chemical formula 2]

(In the formula, R ¹ represents an alkylene group, R ² and R ³ represent an alkenyl group, R ⁴ and R ⁵ represent a phosphocholine residue, and R ⁶ and R ⁷ represent a hydrogen atom or an alkyl group, respectively.)
[0007]
DETAILED DESCRIPTION OF THE INVENTION
The novel sphingolipid derivative that is the target product of the present invention can be represented by the following general formula (1).
General formula (1)
[Chemical 3]

(Each symbol in the formula is the same as above.)
[0008]
R ¹ represents an alkylene group. Examples of the alkylene group include alkylene groups having 1 to 60, preferably 1 to 25, carbon atoms constituting the main chain. Specific examples of such an alkylene group include those obtained by removing one hydrogen atom from an alkyl group described later.
R ² and R ³ represent an alkenyl group. Examples of the alkenyl group include alkenyl groups having 2 to 50, preferably 2 to 25 carbon atoms constituting the main chain. Specific examples of such alkenyl groups include vinyl, propenyl, butenyl, acrylic, methacryl, octynyl, dodecenyl, undecenyl and the like.
R ⁴ and R ⁵ represent phosphocholine residues.
R ⁶ and R ⁷ represent a hydrogen atom or an alkyl group. Examples of the alkyl group include those having 1 to 60 carbon atoms, preferably 1 to 25 carbon atoms constituting the main chain. Examples of such an alkyl group, e.g., methyl, ethyl, n - propyl, isopropyl, n - butyl, isobutyl, s e c- butyl, t - butyl, n - pentyl, isopentyl, 2 - methylbutyl, 1 - methylbutyl, Examples include n- hexyl, isohexyl, 3 -methylpentyl, 2 -methylpentyl, 1 -methylpentyl, heptyl, octyl, isooctyl, 2 -ethylhexyl, nonyl, decyl, undecyl, dodecyl, tetradecyl, hexadecyl, octadecyl, eicosyl, etc. Is done.
[0027]
The sphingolipid derivative represented by the general formula (1) of the present invention can be synthesized, for example, as follows.
First, as shown in Table 1, according to the method described in Non-Patent Document 2, serine or a serine derivative is used as a starting material, and an intermediate Ghana-aldehyde is synthesized. Next, R ² or R ³ is coupled to this Ghana-aldehyde, followed by deacetonide and de-Boc, leading to a sphingosine derivative. Subsequently, R ⁴ or R ⁵ is reacted with a primary hydroxyl group, and then R ⁶ or R ⁷ is reacted with a secondary hydroxyl group to synthesize compounds A and B. Subsequently, a desired sphingolipid derivative can be obtained by coupling the two with R ¹ . In addition, as shown in Table 2 below, the sphingosine derivative and R ¹ are reacted, then R ⁴ or R ⁵ is reacted with a primary hydroxyl group, and then R ⁶ or R ⁷ is reacted with a secondary hydroxyl group to obtain the desired Sphingolipid derivatives can also be obtained.
[0028]
[Table 1]

[0029]
Since the sphingolipid derivative represented by the general formula (1) of the present invention has two ceramide derivatives in the same molecule, the sphingolipid derivative consisting of one ceramide molecule accumulates on the outer membrane and inner membrane of the cell membrane. The positions of the intima and outer membrane are different. A sphingolipid derivative having two ceramide derivatives in the same molecule is thought to exist in the state of penetrating the outer and inner membranes when accumulated in the cell membrane, and has the property of a transmembrane type surface activity. It can be widely used in medicines such as antitumor agents and anti-influenza agents, cosmetics such as moisturizers and hair restorers, agricultural chemicals and industrial chemicals.
[0030]
【Example】
EXAMPLES Hereinafter, although an Example is given and demonstrated about this invention, unless it exceeds the summary, it is not limited to the following.
[0031]
According to the synthesis scheme shown in Table 2, the sphingolipid derivative (Compound 12) of the present invention was synthesized.
[0032]
[Synthesis of Compound 1]
Compound 1 was synthesized with reference to the following literature.
Helv. Chim. Acta. , 1988, 354; Med. Chem. , 1999, 2687, Bioorg. Med. Chem. Lett. , 1998, 179
[0033]
[Synthesis of Compound 2]
Under a nitrogen atmosphere, 1-heptin (3.87 mL, 29.4 mmol) was dissolved in dehydrated tetrahydrofuran and stirred at -40 degrees. To this solution, a hexane solution of n-butyllithium (1 g / mL, 18.9 mL) was added dropwise and stirred at the same temperature for 15 minutes. To this reaction solution, a tetrahydrofuran solution of compound 1 (2.25 g, 9.82 mmol) was added dropwise and slowly heated to −20 ° C. over 1 hour. A saturated aqueous ammonium chloride solution was added to the solution, and then extraction was performed three times with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate. The syrup-like crude product (3.215 g) obtained by distilling off the organic layer under reduced pressure was purified by silica gel column chromatography (n-hexane: ethyl acetate = 10: 1) to give compound 2 (2.3 g). , 72.1%).
[0034]
[Synthesis of Compound 3]
Compound 2 (1.5 g, 4.61 mmol) was dissolved in methanol, p-toluenesulfonic acid monohydrate (10 mg) was added, and the mixture was stirred at room temperature under a nitrogen atmosphere. After 25 hours, p-toluenesulfonic acid monohydrate (20 mg) was added, and the mixture was further stirred for 24 hours. To the residue obtained by distilling off methanol under reduced pressure, a saturated aqueous solution of sodium hydrogen carbonate and ethyl acetate were added for liquid separation. The aqueous layer was further extracted twice with ethyl acetate, and the resulting organic layer was washed with saturated brine and dried over sodium sulfate. A syrupy crude product (1.305 g) obtained by distilling off the organic layer under reduced pressure was purified by silica gel column chromatography (n-hexane: ethyl acetate = 3: 1 to 1: 1) to give compound 3 (1.25 g, 94.9%) was obtained.
[0035]
[Synthesis of Compound 4]
Under a nitrogen atmosphere, the deacetal form (726 mg, 2.55 mmol) was dissolved in dehydrated ethyl ether and stirred at −20 degrees. To this solution, a solution obtained by diluting a toluene solution of Red-Al (concentration 65%, 3.82 mL, 12.7 mmol) with dehydrated ethyl ether was added dropwise, and the mixture was slowly heated from −20 ° C. to room temperature over 1 hour. . The reaction solution was further stirred at room temperature for 24 hours, methanol and a saturated aqueous Rochelle salt solution (50 mL) were added, and the mixture was extracted 3 times with ethyl acetate. The obtained organic layer was washed with saturated brine and dried over magnesium sulfate. The crude product obtained by distilling off the organic layer under reduced pressure was purified by silica gel column chromatography (n-hexane: ethyl acetate = 2: 1 to 1: 2) to give compound 5 (488 mg, 66.7%). Got.
[0036]
[Synthesis of Compound 5]
Compound 4 (485 mg, 1.69 mmol) was dissolved in trifluoroacetic acid (1.0 mL), stirred for 15 minutes under ice cooling, and then stirred at room temperature for 30 minutes. Further, trifluoroacetic acid (0.5 mL) was added, and the mixture was stirred at room temperature for 1 hour. Trifluoroacetic acid was distilled off under reduced pressure, 2M aqueous sodium hydroxide solution (5 mL) was added, and the mixture was extracted 5 times with ethyl ether. The obtained organic layer was dried over sodium sulfate, and ethyl ether was distilled off under reduced pressure to obtain Compound 5 (310 mg, 98%).
[0037]
[Synthesis of Compound 6]
The prepared hexadecanedioic acid succinimide ester (15 mg, 0.05 mmol) was added to a dehydrated tetrahydrofuran solution of compound 5 (19 mg, 0.1 mmol), and the mixture was stirred at room temperature in a nitrogen atmosphere. To this solution was added dehydrated triethylamine (0.014 mL, 0.1 mmol), and the mixture was stirred at room temperature for 24 hours under a nitrogen atmosphere. Thereafter, the solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane: ethyl acetate: methanol = 10: 10: 1) to obtain compound 6 (23.1 mg, 73.9%). Got.
[0038]
The product was identified by ¹ H-NMR spectrum.
[0039]
¹ H-NMR (TMS, CDCl ₃ ): 6.28 (2H, d, J = 7.7 Hz), 5.79 (2H, dtd, J = 15.3, 6.7, 1.0 Hz), 5 .53 (2H, ddt, J = 15.3, 6.4, 1.0 Hz), 4.32 (2H, brs), 3.95 (2H, brd, J = 11.9 Hz), 3.91 ( 2H, m), 3.71 (2H, m), 2.85 (4H, brs), 2.23 (4H, t, J = 7.4 Hz), 2.06 (4H, dt, J = 7. 5, 6.7 Hz), 1.64 (4H, tt, J = 7.4, 7.4 Hz), 1.38 (4H, tt, J = 7.4, 7.4 Hz), 1.25-1 .30 (14H, m), 0.89 (6H, t, J = 7.0 Hz).
[0040]
[Synthesis of Compound 7]
N, N-dimethylaminopyridine (0.032 mg, 0.0026 mmol) was added to an anhydrous pyridine solution of compound 6 (10 mg, 0.016 mmol), and the mixture was stirred at room temperature for 2 hours, and then benzoyl chloride (0.006 ml, 0.006 mmol). 052 mmol) was added and stirred at the same temperature for 20 hours. Saturated aqueous ammonium chloride solution was added and stirred vigorously, followed by extraction three times with chloroform. The obtained organic layer was washed with a saturated aqueous sodium chloride solution and dehydrated with sodium sulfate. The crude product obtained by evaporating the organic layer under reduced pressure was purified by silica gel column chromatography (n-hexane: ethyl acetate = 2: 1 to n-hexane: ethyl acetate: methanol = 6: 6: 1), Compound 7 (7.3 mg, 54.8%) was obtained.
[0041]
The product was identified by ¹ H-NMR spectrum.
[0042]
¹ H-NMR (TMS, CDCl ₃ ): 8.02 (4H, d, J = 8.3 Hz), 7.58 (2H, t, J = 7.6 Hz), 7.45 (4H, t, J = 8.0 Hz), 5.98 (2H, d, J = 7.9 Hz), 5.77 (2H, dt, J = 15.5, 6.7 Hz), 5.53 (2H, ddt, J = 15.5, 6.7, 1.0 Hz), 4.56 (2H, dd, J = 12.5, 8.3 Hz), 4.41 (4H, m), 4.27 (2H, brs) ), 2.87 (2H, d, J = 5.0 Hz), 2.19 (4H, t, J = 7.6 Hz), 2.01 (4H, m), 1.58 (4H, m), 1.34 (4H, m), 1.34-1.20 (28H, m), 0.87 (6H, t, J = 7.0 Hz).
[0043]
[Synthesis of Compound 8]
Under ice cooling, 2,6-lutidine (0.011 ml, 0.096 mmol) was added to a dehydrated dichloromethane solution of compound 7 (20 mg, 0.024 mmol), and the mixture was stirred at the same temperature for 1 hour. To this solution, t-butyldimethylsilane triflate (0.072 mmol) was slowly added dropwise, and then slowly raised to room temperature, and further stirred at room temperature for 1 hour. Saturated brine was added and the mixture was stirred and extracted three times with dichloromethane. The obtained organic layer was dried over sodium sulfate and distilled under reduced pressure to obtain Compound 8.
[0044]
[Synthesis of Compound 9]
To a dehydrated dichloromethane: dehydrated methanol solution (1: 1) of compound 8 (20 mg, 0.024 mmol), potassium carbonate (20 mg) was added and allowed to stand in a refrigerator for 3 days to obtain compound 9 quantitatively.
[0045]
[Synthesis of Compound 10]
To 5 equivalents of 2-bromoethyl dichlorophosphate in anhydrous diethyl ether, an equivalent amount of triethylamine and Compound 9 in anhydrous diethyl ether were added dropwise under ice cooling. After stirring at the same temperature for 3.5 hours, 0.5N aqueous potassium chloride solution was added, diluted with diethyl ether, and stirred for 1 hour. After extraction with diethyl ether, the mixture was washed with water and saturated brine in this order. The separated organic layer was dried over anhydrous magnesium sulfate and filtered, and then the solvent was distilled off under reduced pressure to obtain Compound 10.
[0046]
[Synthesis of Compound 11]
Compound 10 was dissolved in chloroform: isopropyl alcohol: acetonitrile (3: 3: 5), 30% trimethylamine aqueous solution was added, and the mixture was stirred at 50 ° C. for 4 hours. After cooling, the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: methanol = 10: 1) to obtain Compound 11 at 74%.
[0047]
[Synthesis of Compound 12]
To an anhydrous tetrahydrofuran solution of compound 11, 2 equivalents of tetra n-butylammonium fluoride (1.0 M tetrahydrofuran solution) was added under ice cooling, and the mixture was stirred at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform: methanol = 5: 1) to obtain Compound 12 in 88% yield.
[0048]
The product was identified by ¹ H-NMR spectrum.
[0049]
¹ H-NMR (CDCl ₃ -CD ₃ OD, TMS): 7.19 (2H, d, J = 8.3 Hz), 5.64 (2H, dtd, J = 15.3, 6.7, 1. 0 Hz), 5.40 (2H, ddt, J = 15.3, 6.4, 1.0 Hz), 4.32 (2H, brs), 3.92-3.81 (8H, m), 3. 71 (2H, m), 3.43 (4H, m), 3.20 (18H, s), 2.85 (2H, brs), 2.23 (4H, t, J = 7.4 Hz), 2 .06 (4H, dt, J = 7.5, 6.7 Hz), 1.64 (4H, tt, J = 7.4, 7.4 Hz), 1.38 (4H, tt, J = 7.4) 7.4 Hz), 1.25-1.30 (28 H, m), 0.89 (6 H, t, J = 7.0 Hz).
[0050]
[Table 2]

[0051]
【The invention's effect】
According to the present invention, a novel sphingolipid derivative having two ceramide derivatives in the same molecule is provided.
The sphingolipid derivatives obtained by the method of the present invention are widely used in medicines such as antitumor agents and anti-influenza agents, cosmetics such as humectants and hair restorers, agricultural chemicals and industrial chemicals.

Claims (1)

Represented by the following general formula (1),
General formula (1)

(In the formula, R ¹ represents an alkylene group, R ² and R ³ represent an alkenyl group, R ⁴ and R ⁵ represent a phosphocholine residue, and R ⁶ and R ⁷ represent a hydrogen atom or an alkyl group, respectively.)
A sphingolipid derivative represented by: