JP4305010B2 - Phytosphingosine analogues and their production - Google Patents

Description

（式中、Xは炭素数１〜１５を有するアルキル基を意味し、Ｐ¹は水酸基の保護基を意味する）
【０００６】
本発明の化合物（１２）の製造法につき以下に詳細に説明する。
その製造工程は以下のスキーム１で示される。
スキーム１
【化３３】

（上記式中、Xは炭素数１〜１５を有するアルキル基、Zは蛍光標識基、P¹は水酸基の保護基をそれぞれ意味する。またＲ¹およびＲ²は、同一または異なって水素原子、炭素数１〜４のアルキル基、フェニル基を意味し、Ｒ¹およびＲ²は隣接する炭素原子と共に炭素数３〜６のシクロアルキル環を形成してもよい。）
【０００７】
式（１）で表される化合物の炭素炭素二重結合をエポキシ化することにより式（２）で表される化合物が得られる。
エポキシ化に使用する試薬としては、過安息香酸、ｍ−クロロ過安息香酸、モノペルオキシフタル酸、トリフルオロ過酢酸、過酢酸等の過酸、ジメチルジオキシラン等の過酸化物、バナジウム、モリブテン、タングステン、チタン等の金属触媒の存在下でのｔ−ブチルヒドロペルオキシド等が挙げられる。後で述べるが、不斉エポキシ化反応を行う場合、光学活性な酒石酸ジアルキルを用いたシャープレスの不斉エポキシ化反応やサレン−マンガン錯体を用いたジェイコブセンの不斉エポキシ化反応を行えばよい。
使用する溶媒としてはヘキサン、ベンゼン、トルエン等の炭化水素系溶媒、Ｎ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒、酢酸エチル、酢酸ブチル等のエステル系溶媒、テトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライムジエチレングリコールモノメチルエーテル等のエーテル系溶媒、アセトン、メチルエチルケトン、メチルイソブチルケトン等のケトン系溶媒、アセトニトリル等のニトリル系溶媒、ジクロロメタン、１，２−ジクロロエタン等のハロゲン系溶媒、エタノール、メタノール等のアルコール系溶媒、水媒体並びにこれらの混合溶媒等が挙げられる。
反応温度は−７８℃から溶媒の還流温度まででよく、より好ましくは−７８℃から２５℃である。
【０００８】
式（２）で表される化合物のエポキシを位置選択的に開環することにより式（３）で表される化合物が得られる。
反応としては、水素雰囲気下での接触還元、水素化リチウムアルミニウム、水素化ジイソブチルアルミニウム等を用いたヒドリド還元が好ましく挙げられるが、より好ましくは水素化ジイソブチルアルミニウムである。使用する溶媒としては、ヘキサン、ベンゼン、トルエン等の炭化水素系溶媒、ジエチルエーテル、テトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライムジエチレングリコールモノメチルエーテル等のエーテル系溶媒が好ましく挙げられる。
反応温度は、−１００℃から溶媒の還流温度まででよく、より好ましくは−７８℃から２５℃である。
【０００９】
一般式（３）で表される化合物の水酸基保護基Ｐ¹を除去することにより一般式（４）で表される化合物が得られる。保護基に応じて常法により除去できるが、例えば、保護基Ｐ¹がテトラヒドロピラニル基の場合、酸性イオン交換樹脂、ｐ−トルエンスルホン酸、ピリジニウムパラトルエンスルホネート等の有機酸等を用いて脱保護してよい。
使用する溶媒としてはヘキサン、ベンゼン、トルエン等の炭化水素系溶媒、Ｎ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒、酢酸エチル、酢酸ブチル等のエステル系溶媒、テトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライムジエチレングリコールモノメチルエーテル等のエーテル系溶媒、アセトン、メチルエチルケトン、メチルイソブチルケトン等のケトン系溶媒、アセトニトリル等のニトリル系溶媒、ジクロロメタン、１，２−ジクロロエタン等のハロゲン系溶媒、エタノール、メタノール等のアルコール系溶媒、水媒体並びにこれらの混合溶媒等が好ましく挙げられる。
反応温度は０℃から溶媒の還流温度まででよく、より好ましくは０℃から２５℃である。
【００１０】
得られた式（４）で表される化合物の２つの２級水酸基を酸の存在下アセタール化剤で保護することにより式（５）で表される化合物が得られる。
使用する酸としては、ｐ−トルエンスルホン酸、ピリジニウムパラトルエンスルホンネート、カンファースルホン酸等の有機酸、塩酸、硫酸、リン酸等の鉱酸、三フッ化ホウ素等のルイス酸が好ましく挙げられる。
アセタール化剤としては、例えば式（５）においてＲ¹＝Ｒ²＝Ｈの場合はホルムアルデヒドを、Ｒ¹＝Ｒ²＝フェニルの場合はベンゾフェノンを、Ｒ¹、Ｒ²が隣接する炭素と共に６員環を形成する化合物の場合はシクロヘキサノンを用いればよい。また、Ｒ¹＝Ｒ²＝メチルの場合はアセタール化剤としてアセトン、２，２−ジメトキシプロパン、２−メトキシプロペンを使用することが特に好ましい。溶媒としては、ヘキサン、ベンゼン、トルエン等の炭化水素系溶媒、Ｎ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒、酢酸エチル、酢酸ブチル等のエステル系溶媒、テトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライムジエチレングリコールモノメチルエーテル等のエーテル系溶媒、アセトン、メチルエチルケトン、メチルイソブチルケトン等のケトン系溶媒、アセトニトリル等のニトリル系溶媒、ジクロロメタン、１，２−ジクロロエタン等のハロゲン系溶媒等が挙げられ、アセタール化剤を溶媒として用いることもできる。
反応温度は０℃から溶媒の還流温度まででよく、より好ましくは０℃から２５℃である。
【００１１】
式（５）で表される化合物の１級水酸基をスルホニル化またはハロゲン化を行うことで脱離基に変え、続いて金属アジド化合物と反応させることにより式（６）で表される化合物が得られる。
スルホニル化は有機溶剤中、塩基存在下、式（５）で表される化合物とスルホニル化剤とを反応させて行うことができる。
スルホニル化剤としては、トルエンスルホニルクロリド、メタンスルホニルクロリド、無水メタンスルホン酸等が挙げられる。スルホニル化の使用量は基質に対して１当量以上でよく、好ましくは１〜５当量である。
使用する溶媒としてはテトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライム、ジエチルエーテル、ジイソプロピルエーテル、ｔ−ブチルメチルエーテル等のエーテル系溶媒、ジクロロメタン、クロロホルム、四塩化炭素、１，２−ジクロロエタン、クロロベンゼン等のハロゲン化炭化水素系溶媒、Ｎ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒、ベンゼン、トルエン等の芳香族炭化水素系溶媒ならびにこれらの混合溶媒等が挙げられる。
また、使用する塩基としてはトリエチルアミン、エチルジイソプロピルアミン、Ｎ，Ｎ−ジメチルアニリン、Ｎ，Ｎ−ジエチルアニリン、ピリジン、ピコリン、ルチジン、コリジン、４−Ｎ，Ｎ−ジメチルアミノピリジン等の３級アミンが挙げられる。これらは単独で用いてもよいし、混合して用いてもよい。
反応温度は−１００℃から溶媒の還流温度まででよく、より好ましくは−５℃から２５℃である。
【００１２】
ハロゲン化は溶媒中、式（５）で表される化合物とハロゲン化剤とを反応させて行う。ハロゲン化剤としては、三塩化リン、三臭化リン、三ヨウ化リン、五塩化リン、五臭化リン、四ヨウ化二リン、オキシ塩化リン、オキシ臭化リン、ジフェニルトリクロロホスホラン、ジフェニルトリブロモホスホラン、トリフェニルホスフィンジクロロニド、トリフェニルホスフィンジブロモニド、ホスホン酸トリフェニルジクロロニド、ホスホン酸トリフェニルジブロモニド、ホスホン酸トリフェニルジヨードニド等のハロゲン化リン化合物；塩化チオニル、臭化チオニル、塩化スルフリル等のハロゲン化硫黄化合物；ベンジルクロリド−ホスホン酸トリフェニル、ヨウ化メチル−ホスホン酸トリフェニル、四塩化炭素−トリオクチルホスフィン、四塩化炭素−トリフェニルホスフィン、四臭化炭素−トリフェニルホスフィン等の有機ハロゲン化物と有機リン化合物との混合物；塩化Ｎ，Ｎ−ジメチルクロロホルミウム、臭化Ｎ，Ｎ−ジメチルクロロホルミウム等のヴィルスマイヤー試薬等が挙げられる。ハロゲン化剤の使用量は基質に対して１当量以上でよく、好ましくは１〜２当量である。
使用する溶媒としてはハロゲン化剤に不活性な溶媒なら何ら限定されるものではない。例えば、テトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライム、ジエチルエーテル、ジイソプロピルエーテル、ｔ−ブチルメチルエーテル等のエーテル系溶媒、ジクロロメタン、クロロホルム、四塩化炭素、１，２−ジクロロエタン、クロロベンゼン等のハロゲン化炭化水素系溶媒、Ｎ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒、ベンゼン、トルエン等の芳香族炭化水素系溶媒、ペンタン、ヘキサン、ヘプタン、デカン、シクロヘキサン等の脂肪族炭化水素系溶媒ならびにこれらの混合溶媒等が挙げられる。また、ハロゲン化剤そのものを溶媒として使用することもできる。
反応温度は−１００℃から溶媒の還流温度までで、より好ましくは−５℃から２５℃である。
【００１３】
金属アジド化合物との反応であるが、使用する試薬としてはアジ化ナトリウムが挙げられる。
使用する溶媒としてはヘキサン、ベンゼン、トルエン等の炭化水素系溶媒、Ｎ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒、酢酸エチル、酢酸ブチル等のエステル系溶媒、テトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライムジエチレングリコールモノメチルエーテル等のエーテル系溶媒、アセトン、メチルエチルケトン、メチルイソブチルケトン等のケトン系溶媒、アセトニトリル等のニトリル系溶媒、ジクロロメタン、１，２−ジクロロエタン等のハロゲン系溶媒、水媒体並びにこれらの混合溶媒等が挙げられるが、好ましくはＮ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒である。
反応温度は０℃から溶媒の還流温度まででよく、より好ましくは２５℃から溶媒の還流温度である。
【００１４】
式（６）で表される化合物のアミノ基を保護することにより式（７）で表される化合物が得られる。 アミノ基の保護基としては公知のアミノ基の保護基はいずれも使用できるが、好ましくはメトキシカルボニル基、ｔ−ブトキシカルボニル基、ベンジルオキシカルボニル基、エトキシカルボニル基、アセチル基、トリフルオロアセチル基、ベンジル基等が挙げられ、特に好ましいのはｔ−ブトキシカルボニル基である。使用する塩基としては、トリエチルアミン、ジイソプロピルエチルアミン、ピリジン、イミダゾール等の３級アミンが挙げられる。また、４−Ｎ，Ｎ−ジメチルアミノピリジン等の触媒を用いることにより反応時間が短縮される。
使用する溶媒としてはヘキサン、ベンゼン、トルエン等の炭化水素系溶媒、Ｎ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒、酢酸エチル、酢酸ブチル等のエステル系溶媒、テトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライムジエチレングリコールモノメチルエーテル等のエーテル系溶媒、アセトン、メチルエチルケトン、メチルイソブチルケトン等のケトン系溶媒、アセトニトリル等のニトリル系溶媒、ジクロロメタン、１，２−ジクロロエタン等のハロゲン系溶媒、水媒体並びにこれらの混合溶媒等が挙げられるが、好ましくはＮ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒である。
反応温度は−７８℃から溶媒の還流温度まででよく、より好ましくは０℃から２５℃である。
【００１５】
式（７）で表される化合物のオキサゾリジノン環を加水分解して開環することにより式（８）で表される化合物が得られる。用いる塩基としては炭酸ナトリウム、炭酸カリウム、炭酸カルシウム、炭酸セシウム等のアルカリ金属もしくはアルカリ土類金属炭酸塩、水酸化ナトリウム、水酸化カリウム、水酸化カルシウム、水酸化バリウム等のアルカリ金属もしくはアルカリ土類金属水酸化物等が挙げられる。塩基の使用量は、基質に対して１〜３当量、好ましくは１〜１．５当量である。
使用する溶媒としては、例えばメタノール、エタノール、２−プロパノール等のアルコール系溶媒、ヘキサン、ベンゼン、トルエン等の炭化水素系溶媒、Ｎ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒、酢酸エチル、酢酸ブチル等のエステル系溶媒、テトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライムジエチレングリコールモノメチルエーテル等のエーテル系溶媒、アセトン、メチルエチルケトン、メチルイソブチルケトン等のケトン系溶媒、アセトニトリル等のニトリル系溶媒、ジクロロメタン、１，２−ジクロロエタン等のハロゲン系溶媒、水媒体並びにこれらの混合溶媒等が挙げられる。
反応温度は−７８℃から溶媒の還流温度まででよく、より好ましくは０℃から２５℃である。
【００１６】
式（８）で表される化合物のアジドを還元することにより式（９）で表される化合物が得られる。
還元の方法としては水素雰囲気下、溶媒中での接触還元またはトリフェニルホスフィンを用いる方法が挙げられる。メタノール、エタノール、２−プロパノール等のアルコール系溶媒、ヘキサン、ベンゼン、トルエン等の炭化水素系溶媒、Ｎ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒、酢酸エチル、酢酸ブチル等のエステル系溶媒、テトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライムジエチレングリコールモノメチルエーテル等のエーテル系溶媒、アセトン、メチルエチルケトン、メチルイソブチルケトン等のケトン系溶媒、アセトニトリル等のニトリル系溶媒、ジクロロメタン、１，２−ジクロロエタン等のハロゲン系溶媒、水媒体並びにこれらの混合溶媒等が挙げられるが、好ましくはテトラヒドロフラン−水である。反応温度は０℃から溶媒の還流温度まででよく、より好ましくは２５℃から溶媒の還流温度である。
【００１７】
式（９）で表される化合物に蛍光標識基（Ｚ）を含む試薬を反応させることにより式（１０）で表される化合物が得られる。
蛍光標識基を含む試薬としては、基質（９）のアミノ基と反応する置換基（例えばハロゲンなど）を有する蛍光標識基ユニットを含む化合物なら、何ら限定されない。例えば４−クロロ−７−ニトロベンゾ−２−オキサ−１，３−ジアゾール、４−フルオロ−７−ニトロベンゾ−２−オキサ−１，３−ジアゾール、９−ブロモアントラセン、ダンシルクロリド等が挙げられる。
この場合、使用する塩基としては、トリエチルアミン、ジイソプロピルエチルアミン、ピリジン、イミダゾール等の３級アミンが挙げられる。
また、使用する溶媒としては、ヘキサン、ベンゼン、トルエン等の炭化水素系溶媒、Ｎ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒、酢酸エチル、酢酸ブチル等のエステル系溶媒、テトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライムジエチレングリコールモノメチルエーテル等のエーテル系溶媒、アセトン、メチルエチルケトン、メチルイソブチルケトン等のケトン系溶媒、アセトニトリル等のニトリル系溶媒、ジクロロメタン、１，２−ジクロロエタン等のハロゲン系溶媒、水媒体並びにこれらの混合溶媒等が挙げられるが、好ましくはテトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライムジエチレングリコールモノメチルエーテル等のエーテル系溶媒である。
反応温度は０℃から溶媒の還流温度まででよく、より好ましくは０℃から５０℃である。
【００１８】
式（１０）で表される化合物を脱アセトナイドすることにより式（１１）で表される化合物が得られる。
脱アセトナイドは公知の方法で行うことができるが、例えば、メタノール、エタノール等のアルコール系溶媒中、希塩酸や希硫酸等の鉱酸やｐ−トルエンスルホン酸やピリジニウムパラトルエンスルホネート等の有機酸を用いることにより脱アセトナイドすることができる。
反応温度は−７８℃から溶媒の還流温度までで、より好ましくは０℃から５０℃である。
【００１９】
式（１１）で表される化合物の水酸基に近いほうのアミノ基を脱保護することにより、式（１２）で表されるフィトスフィンゴシン類縁体が得られる。
アミノ基の脱保護は保護基に適した方法により行うことができるが、例えばｔ−ブトキシカルボニル基の場合、トリメチルシリルブロミド、トリメチルシリルアイオダイド等を用いることができる。
使用する溶媒としてはヘキサン、ベンゼン、トルエン等の炭化水素系溶媒、Ｎ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒、酢酸エチル、酢酸ブチル等のエステル系溶媒、テトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライムジエチレングリコールモノメチルエーテル等のエーテル系溶媒、アセトン、メチルエチルケトン、メチルイソブチルケトン等のケトン系溶媒、アセトニトリル等のニトリル系溶媒、ジクロロメタン、１，２−ジクロロエタン等のハロゲン系溶媒、エタノール、メタノール等のアルコール系溶媒が挙げられる。
反応温度は０℃から溶媒の還流温度までで、好ましくは０℃から５０℃である。
【００２０】
また、光学活性なフィトスフィンゴミエリン類縁体は、出発物質である上記スキーム１に記載の式（１）で表される化合物に光学活性体を用い、加えて（１）から（２）の反応を不斉エポキシ化することで製造することができる。
スキーム２
【化３４】

（式中、Xは炭素数１〜１５を有するアルキル基を意味し、P¹は水酸基の保護基を意味する。）
【００２１】
式（１ａ）で表される化合物を不斉エポキシ化することにより式（２ａ）で表される化合物が得られる。
不斉エポキシ化の方法としては、先ほども述べたが光学活性な酒石酸ジアルキルを用いたシャープレスの不斉エポキシ化反応やサレン−マンガン錯体を用いたジェイコブセンの不斉エポキシ化反応を行ってよい。式（２ａ）で表される化合物のエポキシを位置選択的に開環することにより式（３ａ）で表される化合物が得られる。
反応としては、水素雰囲気下での接触還元、水素化リチウムアルミニウム、水素化ジイソブチルアルミニウム等を用いたヒドリド還元が挙げられるが、好ましくは水素化ジイソブチルアルミニウムである。
使用する溶媒としては、ヘキサン、ベンゼン、トルエン等の炭化水素系溶媒、ジエチルエーテル、テトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライムジエチレングリコールモノメチルエーテル等のエーテル系溶媒が挙げられる。
反応温度は、−７８℃から溶媒の還流温度まででよく、より好ましくは−７８℃から２５℃である。
以後は上記スキーム１に記載の反応工程により、光学活性なフィトスフィンゴシン類縁体を合成できる。
【００２２】
なお、本発明の対象である式（１２）で表されるフィトスフィンゴシン類縁体には３つの不斉炭素が存在するが、これらの立体構造をＮＭＲにより決定する際、試料を次の操作により処理することができる。
【化３５】

（上記式中、Xは炭素数１〜１５を有するアルキル基、Zは蛍光標識基、P¹は水酸基および１級アミノ基の保護基をそれぞれ意味する。）
【００２３】
つまり、式（１２）で表される化合物の３つの水酸基と１級アミノ基を同時に保護することにより式（１３）で表される化合物が得られる。使用する試薬としては立体が決定できる保護基であれば何ら限定されないが好ましいのはアセチル基である。
使用する試薬としては、アセチルクロリド等のアシルクロリド、無水酢酸等の無水カルボン酸無水物が好ましく挙げられ、特に好ましいのは無水酢酸等の無水カルボン酸無水物である。
使用する溶媒としては、ヘキサン、ベンゼン、トルエン等の炭化水素系溶媒、Ｎ，Ｎ−ジメチルホルムアミド、ジメチルスルホキシド等の非プロトン性極性溶媒、酢酸エチル、酢酸ブチル等のエステル系溶媒、テトラヒドロフラン、１，４−ジオキサン、１，２−ジメトキシエタン、ジグライム、トリグライムジエチレングリコールモノメチルエーテル等のエーテル系溶媒、アセトン、メチルエチルケトン、メチルイソブチルケトン等のケトン系溶媒、アセトニトリル等のニトリル系溶媒、ジクロロメタン、１，２−ジクロロエタン等のハロゲン系溶媒、並びにこれらの混合溶媒等を用いてよい。
使用する塩基としてはトリエチルアミン、エチルジイソプロピルアミン、Ｎ，Ｎ−ジメチルアニリン、Ｎ，Ｎ−ジエチルアニリン、ピリジン、ピコリン、ルチジン、コリジン、４−Ｎ，Ｎ−ジメチルアミノピリジン等の３級アミンが挙げられるが、好ましくはピリジンであり、これは溶媒としても使用できる。
反応温度は、−７８℃から溶媒の還流温度まででよく、より好ましくは０℃から２５℃である。
【００２４】
以下に実施例を示すが、これに限定されるものではない。なお、実施例中の略記号THPはテトラヒドロピラニル基、Ｎ−Ｂｏｃはターシャリーブトキシカルボニル基、NBDは７−ニトロベンゾ−２−オキサ−１，３−ジアゾール基を表す。
【実施例】
実施例１
エポキシド体（２２）の合成
4Å モレキュラーシーブ（2.68 ｇ）、ジクロロメタン(36.3 mL)縣濁液に−２０℃で（＋）−酒石酸ジエチル（0.75 mL, 4.36mmol）、チタニウムテトライソプロポキシド (0.86 mL, 2.90 mmol)を加え１０分間攪拌した後、１．３Ｍ ｔ−ブチルヒドロペルオキシドのトルエン溶液 (22.3mL, 29.0 mmol)、アルコール体（２１）(5.36 ｇ, 14.5 mmol)の塩化メチレン（36.3 mL）溶液を加え徐々に０℃まで昇温した。反応終了後、反応混合物を濾過し、そのろ液に硫酸鉄、クエン酸飽和水溶液を加え０℃で３０分間攪拌した。クロロホルムで抽出を行い、有機層を飽和食塩水で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(from 40% to 67% ethyl acetate in hexane)により分離・精製し、エポキシド体（２２）とそのジアステレオマーの混合物 （８：１） を得た(5.48 g, 97.9%)。
【化３６】

【００２５】
実施例２
ジオール体（２３）の合成
エポキシド（２２）とそのジアステレオマーの混合物（2.25 ｇ, 5.84 mmol）のトルエン(29.2 mL)溶液を−７８℃で１Ｍ ジイソブチルアルムニウムヒドリドのトルエン溶液(29.2 mL, 29.2 mmol)を滴下し徐々に０℃まで昇温した。８時間攪拌した後、２Ｎ塩酸で中和し、反応混合物を酢酸エチルで抽出した。有機層を、飽和炭酸水素ナトリウム水溶液、飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(from 75% to 80% ethyl acetate in hexane)により分離・精製し、ジオール体（２３）とそのジアステレオマーの混合物を得た(1.46 g, 64.6%)。
【化３７】

【００２６】
実施例３
トリオール体（２４）の合成
ジオール（２３）とそのジアステレオマーの混合物(1.46 g, 3.77mmol)のメタノール(18.9 mL)溶液にイオン交換樹脂（ＤｏｗｅＸ ５０Ｗ−Ｘ８、439 mg）を加え、室温で１４時間攪拌した後、反応混合物を濾過後、ろ液を減圧濃縮した。その残さを酢酸エチル−メタノールで再結晶を行い、トリオール体（２４） (383 mg, 33.4％)を得た。
【化３８】

[≡]_D ^24.0 6.00 (c=0.776, CH₃OH)
IR (KBr disk): 3464, 3351, 3285, 2917, 1742, 1699, 1470, 1252, 1074, 1030 cm^-1
1H NMR (CD₃OD, 400MHz) δ: 4.46 (dd, J = 6.10, 8.79 Hz, 1H), 4.39 (dd, J = 8.79, 8.79 Hz, 1H), 4.11 (ddd, J = 2.68, 6.10, 6.10 Hz, 1H), 3.53 (dd, J = 6.83, 6.83 Hz, 1H), 3.41 (m, 2H), 1.68 (m, 1H), 1.50 (m, 1H), 1.32 (m, 14H);
¹³C NMR (CD₃OD, 100MHz) δ:162.73, 75.68, 73.63, 67.22, 63.01, 55.45, 34.77, 33.66, 30.83, 30.74, 30.69, 30.60, 26.94, 26.60 .
【００２７】
実施例４
アセトナイド体（２５）の合成
トリオール体（２４） (140 mg, 0.46 mmol)のＮ，Ｎ−ジメチルホルムアミド(2.3 mL)溶液に０℃で、２，２−ジメトキシプロパン (0.17 mL, 1.38 mmol)、カンファースルホン酸 (54 mg, 0.23 mmol)を加え、室温にて５時間攪拌した。その後反応混合物を０℃に冷やし、水を加え５分間攪拌した。反応混合物に飽和炭酸水素ナトリウム水溶液を加え、中和した後、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(from 50% to 67% ethyl acetate in hexane)により分離・精製し、アセトナイド体（２５） (155 mg, 98%)を得た。
【化３９】

[≡]_D ^24.8 ≡9.47 (c=0.835, CHCl₃)
IR (KBr disk): 3287, 2924, 2857, 1748, 1723, 1397, 1233, 1051 cm^-1
1H NMR (CDCl₃, 400MHz) δ:6.05 (d, J = 13.91 Hz, 1H), 4.45 (dd, J = 8.78, 8.78 Hz, 1H), 4.41 (dd, J = 5.86, 9.03 Hz, 1H), 4.18 (m 1H ), 4.00 (dd, J = 5.85, 7.56 Hz, 1H), 3.89 (m, 1H), 3.64 (t, J = 5.12 Hz, 2H), 1.80 (brd, J = 4.88 Hz,1H), 1.60-1.30 (m, 24H) ;
¹³C NMR (CDCl₃, 100MHz) δ:160.12, 108.36, 79.06, 76.81, 67.71, 62.96, 51.66, 32.71, 29.44, 29.37,29.33, 29.29, 27.74, 26.63, 25.66, 25.24 .
【００２８】
実施例５
アジド体（２６）の合成
アセトナイド体（２５）(792 mg, 2.31 mmol)のテトラヒドロフラン(11.5 mL)溶液に０℃でトリエチルアミン(1.04 mL, 6.92 mmol)、メタンスルホニルクロリド(0.27 mL, 3.46 mmol)を順次加え、２０分間攪拌した。反応混合物に飽和塩化アンモニウム水溶液を加え、中和した後、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣へＮ，Ｎ−ジメチルホルムアミド(11.5 mL)、アジ化ナトリウム(750 mg, 11.5 mmol)を順次加えた後、５０℃に昇温し３時間攪拌した。反応混合物に水を加えた後、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(from 20% to 25% ethyl acetate in hexane)により分離・精製し、アジド体（２６）(738 mg, 86.8%)を得た。
【化４０】

[≡]_D ^24.0 ≡6.94 (c=1.021, CHCl₃)
IR (KBr disk): 3274, 2917, 2857, 2095, 1746, 1719, 1233, 192, 1051 cm^-1
1H NMR (CDCl₃, 400MHz) δ:7.27 (m, 1H), 4.45 (dd, J = 8.30, 9.02 Hz, 1H), 4.41 (dd, J = 5.85, 9.02 Hz, 1H), 4.18 (ddd, J = 5.36, 5.61, 7.66 Hz 1H ), 4.00 (dd, J = 5.85, 7.56 Hz, 1H), 3.89 (m, 1H), 3.26 (t, J = 6.84 Hz, 2H), 1.72-1.29 (m, 24H) ;
¹³C NMR (CDCl₃, 100MHz) δ:160.11, 108.37, 79.09, 76.82, 67.73, 51.67, 51.46, 29.38, 29.32, 29.09, 28.79, 27.76, 26.68, 25.25 .
【００２９】
実施例６
Ｎ−Ｂｏｃ体（２７）の合成
アジド体（２６）(738 mg, 2.00 mmol)のＮ，Ｎ−ジメチルホルムアミド(10.0 mL)溶液に０℃で４−Ｎ，Ｎ−ジメチルアミノピリジン(122 mg, 1.00 mmol)、トリエチルアミン(0.45 mL, 3.00 mmol)、二炭酸ジ−ｔ−ブチル(655 mg, 3.00 mmol)を順次加え、３時間攪拌した。反応混合物に飽和塩化アンモニウム水溶液を加え、中和した後、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(from 11% to 20% ethyl acetate in hexane)により分離・精製し、Ｎ−Ｂｏｃ体（２７）(925 mg, 98.5%)を得た。
【化４１】

[≡]_D ^24.0 ≡63.48 (c=1.042, CHCl₃)
IR (Neat): 3437, 2932, 2857, 2097, 1823, 1798, 1721, 1333, 1260, 1209, 1161, 1080 cm^-1
1H NMR (CDCl₃, 400MHz) δ:4.53 (dd, J =7.32, 7.32 Hz, 1H), 4.45 (dd, J = 6.34, 6.34 Hz, 1H), 4.26 (m 3H ), 3.26 (t, J = 6.83 Hz, 2H), 1.65-1.29 (m, 33H) ;
¹³C NMR (CDCl₃, 100MHz) δ:152.06, 149.60, 108.57, 84.03, 76.28, 75.40, 62.57, 55.69, 51.44, 29.73, 29.46, 29.37, 29.33, 29.08, 28.79, 28.01, 26.83, 26.66, 26.08, 24.50 .
【００３０】
実施例７
アルコール体（２８）の合成
Ｎ−Ｂｏｃ体（２７）(925 mg, 1.97 mmol)のメタノール(0.99 mL)溶液に炭酸セシウム(322 mg, 0.987 mmol)を加え、室温で１．５時間攪拌した。反応混合物に飽和塩化アンモニウム水溶液を加え、中和した後、減圧濃縮を行い、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(from 20% to 25% ethyl acetate in hexane)により分離・精製し、アルコール体（２８）(820 mg, 93.8%)を得た。
【化４２】

[≡]_D ^24.0 9.83 (c=0.543, CHCl₃)
IR (Neat): 3360, 2928, 2930, 2857, 2097, 1698, 1524, 1368, 1248, 1171, 1046 cm^-1
1H NMR (CDCl₃, 400MHz) δ:4.96 (d, J =8.44 Hz, 1H), 4.17 (m, 1H), 4.08 (dd, J = 6.10, 6.10 Hz, 1H), 3.84 (m 1H ), 3.77 (m, 1H), 3.69 (ddd, J =3.66, 7.07, 10.73, 1H), 3.25 (t, J = 6.83 Hz, 2H), 2.33 (br, 1H), 1.67-1.28 (m, 33H) ;
¹³C NMR (CDCl₃, 100MHz) δ:155.39, 108.07, 79.73, 78.17, 77.83, 63.80, 51.47, 51.06, 29.51, 29.41, 29.20, 29.12, 28.81, 28.33, 27.60, 26.68, 25.26 .
【００３１】
実施例８
アミン体（２９）の合成
アルコール体（２８）(794 mg, 1.79 mmol)のテトラヒドロフラン(16.1 mL)溶液に水(1.8 mL)、トリフェニルホスフィン(706 mg, 2.69 mmol)を順次加えた後、６０℃に昇温し3時間攪拌した。反応混合物に水を加えた後、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(from 13% to 25% methanoal in chloroform)により分離・精製し、アミン体（２９）(709 mg, 94.9%)を得た。
【化４３】

[≡]_D ^24.0 8.34 (c=0.873, CHCl₃)
IR (KBr disk): 3362, 2928, 1686, 1528, 1368, 1173 cm^-1
1H NMR (CDCl₃, 400MHz) δ:7.80 (d, J =7.80 Hz, 1H), 4.16 (m, 1H), 4.08 (dd, J = 6.10, 6.10 Hz, 1H), 3.83 (dd, J =2.69, 10.74, 1H ), 3.76 (m, 1H), 3.69 (dd, J =3.17, 10.97, 1H), 2.69 (t, J = 6.83 Hz, 2H), 2.12 (brs, 3H), 1.56-1.28 (m, 33H) ;
¹³C NMR (CDCl₃, 100MHz) δ:155.39, 108.00, 77.93, 77.84, 63.52, 51.09, 42.01, 33.40, 29.50, 29.44, 29.39, 29.14, 28.33, 27.64, 26.80, 26.67, 25.32 .
【００３２】
実施例９
NBD体（３０）の合成
アミン体（２９）(648 mg, 1.56 mmol)のテトラヒドロフラン(7.80 mL)溶液に０℃でトリエチルアミン(0.35 mL, 2.33 mmol)、４−クロロ−７−ニトロベンゾ−２−オキサ−１，３−ジアゾール(466 mg, 2.33 mmol)を順次加えた後、室温に昇温し１時間攪拌した。反応混合物に飽和塩化アンモニウム水溶液を加え、中和した後、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(from 25% to 50% ethyl acetate in hexane)により分離・精製し、NBD体（３０）(666 mg, 73.9%)を得た。
【化４４】

[≡]_D ^25.0 2.70 (c=0.888, CHCl₃)
IR (KBr disk): 3343, 2930, 1698, 1586, 1304, 1169, 1044 cm^-1
1H NMR (CDCl₃, 400MHz) δ:8.49 (d, J =8.78 Hz, 1H), 6.44 (brs, 1H), 6.18 (d, J =8.78 Hz, 1H), 4.98 (d, J =9.27 Hz, 1H), 4.17 (m, 1H), 4.09 (m, 1H), 3.84 (m, 1H ), 3.78 (m, 1H), 3.70 (ddd, J =3.66, 7.07, 10.98, 1H), 3.50 (t, J = 7.07 Hz, 1H), 3.49 (t, J = 7.07 Hz, 1H), 2.39 (brs, 1H), 1.81(ddd, J =7.32, 7.32, 14.64, 2H) 1.57-1.29 (m, 35H) ;
¹³C NMR (CDCl₃, 100MHz) δ:155.45, 144.24, 144.13, 143.94, 136.53, 108.08, 98.47, 79.74, 78.19, 77.81, 63.78, 51.09, 43.99, 29.46, 29.31, 29.20, 29.09, 28.45, 28.32, 27.57, 26.87, 26.66, 25.25 .
【００３３】
実施例１０
脱アセトナイド体（３１）の合成
NBD体（３０）(578 mg, 1.00 mmol)のメタノール(5.00 mL)溶液に０℃で２Ｎ塩酸水溶液(2.00 mL)を加えた後、室温に昇温し３．５時間攪拌した。反応混合物に飽和炭酸水素ナトリウム水溶液を加え、中和した後、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(from 50% to 75% ethyl acetate in hexane)により分離・精製し、脱アセトナイド体（３１）(446 mg, 82.9%)を得た。
【化４５】

[≡]_D ^25.0 6.11 (c=0892, CHCl₃)
IR (KBr disk): 3339, 2928, 1688, 1588, 1304, 1171cm^-1
1H NMR (CDCl₃, 400MHz) δ:8.48 (d, J =8.79 Hz, 1H), 6.69 (brs, 1H), 6.18 (d, J = 8.79 Hz, 1H), 5.43 (d, J = 8.30 Hz, 1H), 3.89-3.65 (m, 5H), 3.51 (t, J = 6.35 Hz, 1H), 3.50 (t, J =6.59, 1H), 3.31 (brs,1H), 1.94 (brs, 2H), 1.81 (m, 2H), 1.68-1.28 (m, 25H) ;
¹³C NMR (CDCl₃, 100MHz) δ:156.46, 144.25, 144.11, 143.93, 136.66, 123.52, 98.53, 80.10, 76.18, 72.89, 61.90, 52.90, 44.03, 32.98, 29.39, 29.32, 29.23, 29.21, 29.03, 28.34, 26.82, 25.73 .
【００３４】
実施例１１
NBD−フィトスフィンゴシン（３２）の合成
脱アセトナイド体（３１）(230 mg, 0.426 mmol)のメタノール(2.1 mL)溶液に、室温でトリメチルシリルブロミド (0.28 mL, 2.13 mmol)を滴下し、1.5時間攪拌した。反応混合物を減圧濃縮し、残渣を逆相HPLCにより分離・精製し、NBD−フィトスフィンゴシン（３２）(171 mg, 88.6%)を得た。
【化４６】

[≡]_D ^25.0 ≡1.79 (c=0.699, CH₃OH)
IR (KBr disk): 3341, 2926, 2855, 1588, 1298, 1269 cm^-1
1H NMR (CD₃OD, 400MHz) δ: 8.49 (d, J = 8.78 Hz, 1H), 6.33 (d, J = 9.03 Hz, 1H), 3.94 (dd, J = 3.42, 11.47 Hz, 1H), 3.77 (dd, J = 8.54, 11.47 Hz, 1H), 3.57-3.44 (m, 5H), 1.81-1.32 (m, 18H);
¹³C NMR (CD₃OD, 100MHz) δ:146.66, 145.79, 145.53, 138.56, 122.77, 99.59, 73.37, 73.36, 58.82, 56.45, 44.84, 35.38, 30.77, 30.69, 30.60, 30.34, 29.28, 28.04, 26.23 .
【００３５】
実施例１２
テトラアセチル体（３３）の合成
NBD−フィトスフィンゴシン（３２）(49 mg, 0.112 mmol)のピリジン(0.56 mL)溶液に０℃で無水酢酸(0.22 mL)を加えた後、室温に昇温し3時間攪拌した。飽和硫酸銅水溶液を加えた後、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(from 25% to 20% ethyl acetate in hexane)により分離・精製し、テトラアセチル体（３３）(50 mg, 91.2% for 2steps)を得た。
【化４７】

[≡]_D ^24.5 17.33 (c=0.745, CHCl₃)
IR (Neat): 3322, 2930, 1742, 1586, 1302, 1252, 1046 cm^-1
1H NMR (CDCl₃, 400MHz) δ: 8.49 (d, J = 8.54 Hz, 1H), 6.54 (t, J = 4.88 Hz, 1H), 6.18 (d, J = 8.54 Hz, 1H), 6.03 (d, J = 9.52 Hz, 1H), 5.11 (dd, J = 3.17, 8.05 Hz 1H), 4.95 (dt, J = 3.17, 9.76 Hz, 1H), 4.49 (m, 1H), 4.30 (dd, J = 4.88, 11.46 Hz 1H), 4.03 (dd, J = 3.17, 11.71 Hz 1H), 3.51 (t, J = 6.83 Hz 1H), 3.49 (t, J = 7.07 Hz 1H), 2.08 (s, 3H), 2.05 (s, 6H), 2.02 (s, 3H) 1.20-1.85 (m, 18H);
¹³C NMR (CDCl₃, 100MHz) δ: 98.8, 67.6, 63.0, 62.3, 32.8, 30.7, 29.7, 29.5, 29.45, 29.40, 29.3, 26.2, 25.7, 25.5, 19.6.[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a phytosphingosine analog necessary for elucidating the intracellular behavior of sphingolipids involved in various in vivo information transmission such as protein kinase C inhibition, cell proliferation and cell differentiation, and a method for producing the same.
[0002]
[Background Art and Problems to be Solved by the Invention]
Sphingolipid, a sphingomyelin metabolite, is attracting a great deal of attention because it is involved in various in vivo signal transduction such as cell death, cell differentiation, and protein kinase C inhibition. And many researchers are actively investigating the phenomena related to these lipids. One of the sphingolipids, phytosphingosine, is mainly present in yeast cells and corresponds to sphingosine which is present in mammalian cells and is known as a signal molecule, and many studies have been conducted. However, information on the location of phytosphingosine in the cell and the pathway leading to the phenomenon has not been sufficiently obtained.
At present, as a mechanism for elucidating this metabolic mechanism, compounds such as C6-NBD phytosphingosine and C12-NBD phytosphingosine are commercially available, and this has a fluorescent labeling unit in the acyl group which is a sub-chain. Since it is cleaved by the hydrolase, it is not useful for elucidating the intracellular location of phytosphingosine-1-phosphate in which phytosphingosine is metabolized and the behavior of the metabolite.
[0003]
[Means for Solving the Problems]
Therefore, the present inventors have succeeded in synthesizing a novel phytosphingosine analog, which is a compound having a fluorescent labeling group introduced into the main chain not subjected to metabolism represented by the following formula (12).
Further, using a compound represented by the formula (1) easily obtained from 3-benzyl-4-alkoxycarbonyl-2-oxazolidinone represented by the following formula (14) disclosed in JP-A-10-87777 as a raw material, It has been found that the phytosphingosine analog of the present invention can be synthesized efficiently.
[0004]
When synthesizing an optically active compound, it is important that the operation is simple, the yield is good, and the optical purity is kept high. As a production method meeting such a demand, there is a method of synthesizing a chiral intermediate that is easily chemically converted, and producing a target product through this compound. The important point in this method is that the chiral intermediate is easy to handle in terms of operation, and is inexpensive and available in large quantities. The compounds according to the present invention often exist as optical isomers, and the method of the present invention is a method that satisfies these needs.
[0005]
That is, the present invention is to provide a phytosphingosine analog having a fluorescent labeling group moiety and a method for producing the same, particularly an efficient method.
Embedded image

(In the formula, X means an alkyl group having 1 to 15 carbon atoms, P¹Means a hydroxyl protecting group)
[0006]
The production method of the compound (12) of the present invention will be described in detail below.
The manufacturing process is shown in Scheme 1 below.
Scheme 1
Embedded image

(In the above formula, X is an alkyl group having 1 to 15 carbon atoms, Z is a fluorescent labeling group, P¹Means a hydroxyl-protecting group. Also R¹And R²Are the same or different and each represents a hydrogen atom, an alkyl group having 1 to 4 carbon atoms, or a phenyl group, and R¹And R²May form a cycloalkyl ring having 3 to 6 carbon atoms with adjacent carbon atoms. )
[0007]
A compound represented by the formula (2) is obtained by epoxidizing the carbon-carbon double bond of the compound represented by the formula (1).
Reagents used for epoxidation include perbenzoic acid, m-chloroperbenzoic acid, monoperoxyphthalic acid, trifluoroperacetic acid, peracetic acid such as peracetic acid, peroxides such as dimethyldioxirane, vanadium, molybdenum, And t-butyl hydroperoxide in the presence of a metal catalyst such as tungsten or titanium. As will be described later, when an asymmetric epoxidation reaction is performed, a sharp epoxidation asymmetric epoxidation reaction using optically active dialkyl tartrate or a Jacobsen asymmetric epoxidation reaction using a salen-manganese complex may be performed. .
Solvents used include hydrocarbon solvents such as hexane, benzene and toluene, aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide, ester solvents such as ethyl acetate and butyl acetate, tetrahydrofuran, 1,4 -Ether solvents such as dioxane, 1,2-dimethoxyethane, diglyme, triglyme diethylene glycol monomethyl ether, ketone solvents such as acetone, methyl ethyl ketone, methyl isobutyl ketone, nitrile solvents such as acetonitrile, dichloromethane, 1,2-dichloroethane And halogen solvents such as ethanol, alcohol solvents such as ethanol and methanol, aqueous media, and mixed solvents thereof.
The reaction temperature may be from -78 ° C to the reflux temperature of the solvent, more preferably from -78 ° C to 25 ° C.
[0008]
A compound represented by the formula (3) is obtained by regioselectively opening the epoxy of the compound represented by the formula (2).
Preferred examples of the reaction include catalytic reduction under a hydrogen atmosphere, hydride reduction using lithium aluminum hydride, diisobutylaluminum hydride, and the like, and more preferred is diisobutylaluminum hydride. As the solvent to be used, hydrocarbon solvents such as hexane, benzene and toluene, ether solvents such as diethyl ether, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, diglyme and triglyme diethylene glycol monomethyl ether are preferable. Can be mentioned.
The reaction temperature may be from −100 ° C. to the reflux temperature of the solvent, more preferably from −78 ° C. to 25 ° C.
[0009]
Hydroxyl protecting group P of the compound represented by formula (3)¹Is removed to obtain the compound represented by the general formula (4). Depending on the protecting group, it can be removed by conventional methods, for example protecting group P¹When is a tetrahydropyranyl group, it may be deprotected using an organic acid such as an acidic ion exchange resin, p-toluenesulfonic acid, pyridinium paratoluenesulfonate, or the like.
Solvents used include hydrocarbon solvents such as hexane, benzene and toluene, aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide, ester solvents such as ethyl acetate and butyl acetate, tetrahydrofuran, 1,4 -Ether solvents such as dioxane, 1,2-dimethoxyethane, diglyme, triglyme diethylene glycol monomethyl ether, ketone solvents such as acetone, methyl ethyl ketone, methyl isobutyl ketone, nitrile solvents such as acetonitrile, dichloromethane, 1,2-dichloroethane Preferable examples include halogen solvents such as ethanol, alcohol solvents such as ethanol and methanol, aqueous media, and mixed solvents thereof.
The reaction temperature may be from 0 ° C to the reflux temperature of the solvent, more preferably from 0 ° C to 25 ° C.
[0010]
A compound represented by the formula (5) is obtained by protecting the two secondary hydroxyl groups of the compound represented by the formula (4) with an acetalizing agent in the presence of an acid.
Preferable examples of the acid used include organic acids such as p-toluenesulfonic acid, pyridinium paratoluenesulfonate, and camphorsulfonic acid, mineral acids such as hydrochloric acid, sulfuric acid, and phosphoric acid, and Lewis acids such as boron trifluoride.
As an acetalizing agent, for example, R in formula (5)¹= R²= H, formaldehyde, R¹= R²= Benzophenone in the case of phenyl, R¹, R²In the case of a compound that forms a 6-membered ring with adjacent carbon, cyclohexanone may be used. R¹= R²In the case of = methyl, it is particularly preferable to use acetone, 2,2-dimethoxypropane or 2-methoxypropene as the acetalizing agent. Examples of the solvent include hydrocarbon solvents such as hexane, benzene and toluene, aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide, ester solvents such as ethyl acetate and butyl acetate, tetrahydrofuran, 1,4- Ether solvents such as dioxane, 1,2-dimethoxyethane, diglyme, triglyme diethylene glycol monomethyl ether, ketone solvents such as acetone, methyl ethyl ketone, methyl isobutyl ketone, nitrile solvents such as acetonitrile, dichloromethane, 1,2-dichloroethane, etc. And an acetalizing agent can also be used as a solvent.
The reaction temperature may be from 0 ° C to the reflux temperature of the solvent, more preferably from 0 ° C to 25 ° C.
[0011]
The primary hydroxyl group of the compound represented by the formula (5) is converted into a leaving group by sulfonylation or halogenation, and subsequently reacted with a metal azide compound to obtain the compound represented by the formula (6). It is done.
The sulfonylation can be performed by reacting the compound represented by the formula (5) with a sulfonylating agent in an organic solvent in the presence of a base.
Examples of the sulfonylating agent include toluenesulfonyl chloride, methanesulfonyl chloride, methanesulfonic anhydride and the like. The amount of sulfonylation to be used may be 1 equivalent or more, preferably 1 to 5 equivalents relative to the substrate.
Solvents used include ether solvents such as tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, diglyme, triglyme, diethyl ether, diisopropyl ether, t-butyl methyl ether, dichloromethane, chloroform, carbon tetrachloride, 1 Halogenated hydrocarbon solvents such as 1,2-dichloroethane and chlorobenzene, aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide, aromatic hydrocarbon solvents such as benzene and toluene, and mixed solvents thereof Can be mentioned.
The base used is tertiary amine such as triethylamine, ethyldiisopropylamine, N, N-dimethylaniline, N, N-diethylaniline, pyridine, picoline, lutidine, collidine, 4-N, N-dimethylaminopyridine and the like. Can be mentioned. These may be used alone or in combination.
The reaction temperature may be from −100 ° C. to the reflux temperature of the solvent, more preferably from −5 ° C. to 25 ° C.
[0012]
The halogenation is performed by reacting the compound represented by the formula (5) with a halogenating agent in a solvent. As halogenating agents, phosphorus trichloride, phosphorus tribromide, phosphorus triiodide, phosphorus pentachloride, phosphorus pentabromide, phosphorus tetraiodide, phosphorus oxychloride, phosphorus oxybromide, diphenyltrichlorophosphorane, diphenyl Phosphorus halide compounds such as tribromophosphorane, triphenylphosphine dichloronide, triphenylphosphine dibromonide, phosphonic acid triphenyl dichloronide, phosphonic acid triphenyldibromonide, phosphonic acid triphenyldiiodonide; thionyl chloride, bromide Halogenated sulfur compounds such as thionyl, sulfuryl chloride; benzyl chloride-triphenyl phosphonate, methyl iodide-triphenyl phosphonate, carbon tetrachloride-trioctylphosphine, carbon tetrachloride-triphenylphosphine, carbon tetrabromide-tri Organic halo such as phenylphosphine Mixture of emission product and the organic phosphorus compound; chloride N, N- dimethyl chloro holmium bromide N, Vilsmeier reagent such as N- dimethylchlorosilane holmium and the like. The amount of the halogenating agent used may be 1 equivalent or more, preferably 1 to 2 equivalents relative to the substrate.
The solvent used is not limited as long as it is inert to the halogenating agent. For example, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, diglyme, triglyme, diethyl ether, diisopropyl ether, t-butyl methyl ether and other ether solvents, dichloromethane, chloroform, carbon tetrachloride, 1,2- Halogenated hydrocarbon solvents such as dichloroethane and chlorobenzene, aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide, aromatic hydrocarbon solvents such as benzene and toluene, pentane, hexane, heptane, decane and cyclohexane And aliphatic hydrocarbon solvents such as these and mixed solvents thereof. Moreover, the halogenating agent itself can also be used as a solvent.
The reaction temperature is from −100 ° C. to the reflux temperature of the solvent, more preferably from −5 ° C. to 25 ° C.
[0013]
Although it is reaction with a metal azide compound, sodium azide is mentioned as a reagent to be used.
Solvents used include hydrocarbon solvents such as hexane, benzene and toluene, aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide, ester solvents such as ethyl acetate and butyl acetate, tetrahydrofuran, 1,4 -Ether solvents such as dioxane, 1,2-dimethoxyethane, diglyme, triglyme diethylene glycol monomethyl ether, ketone solvents such as acetone, methyl ethyl ketone, methyl isobutyl ketone, nitrile solvents such as acetonitrile, dichloromethane, 1,2-dichloroethane And a halogen solvent such as N, N-dimethylformamide, and dimethyl sulfoxide. Preferred are aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide.
The reaction temperature may be from 0 ° C. to the reflux temperature of the solvent, more preferably from 25 ° C. to the reflux temperature of the solvent.
[0014]
A compound represented by the formula (7) is obtained by protecting the amino group of the compound represented by the formula (6). Any known protecting group for amino group can be used as the protecting group for amino group, but preferably methoxycarbonyl group, t-butoxycarbonyl group, benzyloxycarbonyl group, ethoxycarbonyl group, acetyl group, trifluoroacetyl group, Examples thereof include a benzyl group, and a t-butoxycarbonyl group is particularly preferable. Examples of the base to be used include tertiary amines such as triethylamine, diisopropylethylamine, pyridine, and imidazole. Further, the reaction time is shortened by using a catalyst such as 4-N, N-dimethylaminopyridine.
Solvents used include hydrocarbon solvents such as hexane, benzene and toluene, aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide, ester solvents such as ethyl acetate and butyl acetate, tetrahydrofuran, 1,4 -Ether solvents such as dioxane, 1,2-dimethoxyethane, diglyme, triglyme diethylene glycol monomethyl ether, ketone solvents such as acetone, methyl ethyl ketone, methyl isobutyl ketone, nitrile solvents such as acetonitrile, dichloromethane, 1,2-dichloroethane And a halogen solvent such as N, N-dimethylformamide, and dimethyl sulfoxide. Preferred are aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide.
The reaction temperature may be from -78 ° C to the reflux temperature of the solvent, more preferably from 0 ° C to 25 ° C.
[0015]
Hydrolysis of the oxazolidinone ring of the compound represented by formula (7) to open the ring yields a compound represented by formula (8). Bases used include alkali metals or alkaline earth metal carbonates such as sodium carbonate, potassium carbonate, calcium carbonate and cesium carbonate, alkali metals or alkaline earth metals such as sodium hydroxide, potassium hydroxide, calcium hydroxide and barium hydroxide. A metal hydroxide etc. are mentioned. The usage-amount of a base is 1-3 equivalent with respect to a substrate, Preferably it is 1-1.5 equivalent.
Examples of the solvent used include alcohol solvents such as methanol, ethanol and 2-propanol, hydrocarbon solvents such as hexane, benzene and toluene, aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide, and acetic acid. Ester solvents such as ethyl and butyl acetate, ether solvents such as tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, diglyme and triglyme diethylene glycol monomethyl ether, and ketone solvents such as acetone, methyl ethyl ketone, and methyl isobutyl ketone Nitrile solvents such as acetonitrile, halogen solvents such as dichloromethane and 1,2-dichloroethane, aqueous media, and mixed solvents thereof.
The reaction temperature may be from -78 ° C to the reflux temperature of the solvent, more preferably from 0 ° C to 25 ° C.
[0016]
The compound represented by the formula (9) is obtained by reducing the azide of the compound represented by the formula (8).
Examples of the reduction method include catalytic reduction in a solvent under hydrogen atmosphere or a method using triphenylphosphine. Alcohol solvents such as methanol, ethanol and 2-propanol, hydrocarbon solvents such as hexane, benzene and toluene, aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide, esters such as ethyl acetate and butyl acetate Solvents, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, diglyme, triglyme diethylene glycol monomethyl ether and other ether solvents, acetone, methyl ethyl ketone, methyl isobutyl ketone and other nitrile solvents, acetonitrile and the like And halogen solvents such as dichloromethane and 1,2-dichloroethane, aqueous media, and mixed solvents thereof, among which tetrahydrofuran-water is preferred. The reaction temperature may be from 0 ° C. to the reflux temperature of the solvent, more preferably from 25 ° C. to the reflux temperature of the solvent.
[0017]
The compound represented by the formula (10) is obtained by reacting the compound represented by the formula (9) with a reagent containing a fluorescent labeling group (Z).
The reagent containing a fluorescent labeling group is not limited as long as it is a compound containing a fluorescent labeling group unit having a substituent (for example, halogen) that reacts with the amino group of the substrate (9). Examples include 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole, 4-fluoro-7-nitrobenzo-2-oxa-1,3-diazole, 9-bromoanthracene, dansyl chloride, and the like.
In this case, examples of the base to be used include tertiary amines such as triethylamine, diisopropylethylamine, pyridine, and imidazole.
Examples of the solvent used include hydrocarbon solvents such as hexane, benzene and toluene, aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide, ester solvents such as ethyl acetate and butyl acetate, tetrahydrofuran, Ether solvents such as 1,4-dioxane, 1,2-dimethoxyethane, diglyme, triglyme diethylene glycol monomethyl ether, ketone solvents such as acetone, methyl ethyl ketone and methyl isobutyl ketone, nitrile solvents such as acetonitrile, dichloromethane, Examples thereof include halogen solvents such as 2-dichloroethane, aqueous media, and mixed solvents thereof. Tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, diglyme, triglyme diethyleneglycol are preferable. An ether solvents such as monomethyl ether.
The reaction temperature may be from 0 ° C to the reflux temperature of the solvent, more preferably from 0 ° C to 50 ° C.
[0018]
The compound represented by the formula (11) is obtained by deacetonating the compound represented by the formula (10).
Deacetonide can be performed by a known method. For example, in an alcohol solvent such as methanol or ethanol, a mineral acid such as dilute hydrochloric acid or dilute sulfuric acid, or an organic acid such as p-toluenesulfonic acid or pyridinium paratoluenesulfonate is used. Can be deacetonated.
The reaction temperature is from -78 ° C to the reflux temperature of the solvent, more preferably from 0 ° C to 50 ° C.
[0019]
By deprotecting the amino group closer to the hydroxyl group of the compound represented by the formula (11), the phytosphingosine analog represented by the formula (12) is obtained.
Deprotection of the amino group can be carried out by a method suitable for the protecting group. For example, in the case of a t-butoxycarbonyl group, trimethylsilyl bromide, trimethylsilyl iodide, or the like can be used.
Solvents used include hydrocarbon solvents such as hexane, benzene and toluene, aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide, ester solvents such as ethyl acetate and butyl acetate, tetrahydrofuran, 1,4 -Ether solvents such as dioxane, 1,2-dimethoxyethane, diglyme, triglyme diethylene glycol monomethyl ether, ketone solvents such as acetone, methyl ethyl ketone, methyl isobutyl ketone, nitrile solvents such as acetonitrile, dichloromethane, 1,2-dichloroethane And halogen solvents such as ethanol and alcohol solvents such as ethanol and methanol.
The reaction temperature is from 0 ° C to the reflux temperature of the solvent, preferably from 0 ° C to 50 ° C.
[0020]
In addition, the optically active phytosphingomyelin analog is obtained by using the optically active substance for the compound represented by the formula (1) described in the above-mentioned scheme 1, which is the starting material, and additionally performing the reactions (1) to (2). It can be produced by asymmetric epoxidation.
Scheme 2
Embedded image

(In the formula, X means an alkyl group having 1 to 15 carbon atoms, P¹Means a hydroxyl-protecting group. )
[0021]
A compound represented by the formula (2a) is obtained by asymmetric epoxidation of the compound represented by the formula (1a).
As described above, the method of asymmetric epoxidation may be performed by sharp pressing asymmetric epoxidation reaction using optically active dialkyl tartrate or a Jacobsen asymmetric epoxidation reaction using salen-manganese complex. . A compound represented by the formula (3a) is obtained by regioselectively opening the epoxy of the compound represented by the formula (2a).
Examples of the reaction include catalytic reduction under a hydrogen atmosphere, hydride reduction using lithium aluminum hydride, diisobutylaluminum hydride, and the like, preferably diisobutylaluminum hydride.
Examples of the solvent used include hydrocarbon solvents such as hexane, benzene, and toluene, and ether solvents such as diethyl ether, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, diglyme, and triglyme diethylene glycol monomethyl ether. It is done.
The reaction temperature may be from -78 ° C to the reflux temperature of the solvent, more preferably from -78 ° C to 25 ° C.
Thereafter, an optically active phytosphingosine analog can be synthesized by the reaction step described in Scheme 1 above.
[0022]
There are three asymmetric carbons in the phytosphingosine analog represented by formula (12), which is the subject of the present invention. When these steric structures are determined by NMR, the sample is treated by the following operation. can do.
Embedded image

(In the above formula, X is an alkyl group having 1 to 15 carbon atoms, Z is a fluorescent labeling group, P¹Means a protective group for a hydroxyl group and a primary amino group, respectively. )
[0023]
That is, the compound represented by the formula (13) is obtained by simultaneously protecting the three hydroxyl groups and the primary amino group of the compound represented by the formula (12). The reagent to be used is not particularly limited as long as it is a protecting group capable of determining a steric structure, but an acetyl group is preferred.
Preferred examples of the reagent to be used include acyl chlorides such as acetyl chloride and carboxylic anhydrides such as acetic anhydride, and particularly preferred are carboxylic anhydrides such as acetic anhydride.
Solvents used include hydrocarbon solvents such as hexane, benzene and toluene, aprotic polar solvents such as N, N-dimethylformamide and dimethyl sulfoxide, ester solvents such as ethyl acetate and butyl acetate, tetrahydrofuran, Ether solvents such as 4-dioxane, 1,2-dimethoxyethane, diglyme and triglyme diethylene glycol monomethyl ether, ketone solvents such as acetone, methyl ethyl ketone and methyl isobutyl ketone, nitrile solvents such as acetonitrile, dichloromethane, 1,2- A halogen-based solvent such as dichloroethane, a mixed solvent thereof or the like may be used.
Examples of the base used include tertiary amines such as triethylamine, ethyldiisopropylamine, N, N-dimethylaniline, N, N-diethylaniline, pyridine, picoline, lutidine, collidine, and 4-N, N-dimethylaminopyridine. Is preferably pyridine, which can also be used as a solvent.
The reaction temperature may be from -78 ° C to the reflux temperature of the solvent, more preferably from 0 ° C to 25 ° C.
[0024]
Although an Example is shown below, it is not limited to this. In the examples, the abbreviation THP represents a tetrahydropyranyl group, N-Boc represents a tertiary butoxycarbonyl group, and NBD represents a 7-nitrobenzo-2-oxa-1,3-diazole group.
【Example】
Example 1
Synthesis of epoxide (22)
4 To a suspension of molecular sieve (2.68 g) and dichloromethane (36.3 mL), add (+)-diethyl tartrate (0.75 mL, 4.36 mmol) and titanium tetraisopropoxide (0.86 mL, 2.90 mmol) at −20 ° C. 10 After stirring for 1 minute, a solution of 1.3M t-butyl hydroperoxide in toluene (22.3 mL, 29.0 mmol) and a solution of alcohol (21) (5.36 g, 14.5 mmol) in methylene chloride (36.3 mL) was gradually added to 0 ° C. The temperature was raised to. After completion of the reaction, the reaction mixture was filtered, and iron sulfate and a saturated aqueous solution of citric acid were added to the filtrate, followed by stirring at 0 ° C. for 30 minutes. Extraction was performed with chloroform, and the organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography (from 40% to 67% ethyl acetate in hexane) to obtain a mixture (8: 1) of epoxide (22) and its diastereomer (5.48 g, 97.9%) .
Embedded image

[0025]
Example 2
Synthesis of diol (23)
A toluene solution (29.2 mL) of a mixture of the epoxide (22) and its diastereomer (2.25 g, 5.84 mmol) was slowly added dropwise with a 1M diisobutylalumnium hydride solution in toluene (29.2 mL, 29.2 mmol) at -78 ° C. The temperature was raised to 0 ° C. After stirring for 8 hours, the mixture was neutralized with 2N hydrochloric acid, and the reaction mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium hydrogen carbonate solution and a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography (from 75% to 80% ethyl acetate in hexane) to obtain a mixture of the diol (23) and its diastereomer (1.46 g, 64.6%).
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[0026]
Example 3
Synthesis of triol body (24)
An ion exchange resin (DoweX 50W-X8, 439 mg) was added to a solution of diol (23) and its diastereomer (1.46 g, 3.77 mmol) in methanol (18.9 mL), and the mixture was stirred at room temperature for 14 hours. After the mixture was filtered, the filtrate was concentrated under reduced pressure. The residue was recrystallized from ethyl acetate-methanol to obtain a triol form (24) (383 mg, 33.4%).
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[≡]_D ^24.06.00 (c = 0.776, CH_ThreeOH)
IR (KBr disk): 3464, 3351, 3285, 2917, 1742, 1699, 1470, 1252, 1074, 1030 cm^-1
1H NMR (CD_ThreeOD, 400MHz) δ: 4.46 (dd, J = 6.10, 8.79 Hz, 1H), 4.39 (dd, J = 8.79, 8.79 Hz, 1H), 4.11 (ddd, J = 2.68, 6.10, 6.10 Hz, 1H), 3.53 (dd, J = 6.83, 6.83 Hz, 1H), 3.41 (m, 2H), 1.68 (m, 1H), 1.50 (m, 1H), 1.32 (m, 14H);
¹³C NMR (CD_Three(OD, 100MHz) δ: 162.73, 75.68, 73.63, 67.22, 63.01, 55.45, 34.77, 33.66, 30.83, 30.74, 30.69, 30.60, 26.94, 26.60.
[0027]
Example 4
Synthesis of acetonide (25)
To a solution of triol form (24) (140 mg, 0.46 mmol) in N, N-dimethylformamide (2.3 mL) at 0 ° C., 2,2-dimethoxypropane (0.17 mL, 1.38 mmol), camphorsulfonic acid (54 mg, 0.23 mmol) was added and stirred at room temperature for 5 hours. Thereafter, the reaction mixture was cooled to 0 ° C., water was added, and the mixture was stirred for 5 minutes. Saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture for neutralization, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography (from 50% to 67% ethyl acetate in hexane) to obtain an acetonide compound (25) (155 mg, 98%).
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[≡]_D ^24.8≡9.47 (c = 0.835, CHCl_Three)
IR (KBr disk): 3287, 2924, 2857, 1748, 1723, 1397, 1233, 1051 cm^-1
1H NMR (CDCl_Three, 400MHz) δ: 6.05 (d, J = 13.91 Hz, 1H), 4.45 (dd, J = 8.78, 8.78 Hz, 1H), 4.41 (dd, J = 5.86, 9.03 Hz, 1H), 4.18 (m 1H) , 4.00 (dd, J = 5.85, 7.56 Hz, 1H), 3.89 (m, 1H), 3.64 (t, J = 5.12 Hz, 2H), 1.80 (brd, J = 4.88 Hz, 1H), 1.60-1.30 ( m, 24H);
¹³C NMR (CDCl_Three, 100MHz) δ: 160.12, 108.36, 79.06, 76.81, 67.71, 62.96, 51.66, 32.71, 29.44, 29.37, 29.33, 29.29, 27.74, 26.63, 25.66, 25.24.
[0028]
Example 5
Synthesis of azide (26)
Triethylamine (1.04 mL, 6.92 mmol) and methanesulfonyl chloride (0.27 mL, 3.46 mmol) were sequentially added to a solution of acetonide compound (25) (792 mg, 2.31 mmol) in tetrahydrofuran (11.5 mL) at 0 ° C., and the mixture was stirred for 20 minutes. . A saturated aqueous ammonium chloride solution was added to the reaction mixture for neutralization, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. N, N-dimethylformamide (11.5 mL) and sodium azide (750 mg, 11.5 mmol) were sequentially added to the residue, and then the mixture was heated to 50 ° C. and stirred for 3 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography (from 20% to 25% ethyl acetate in hexane) to obtain an azide compound (26) (738 mg, 86.8%).
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[≡]_D ^24.0≡6.94 (c = 1.021, CHCl_Three)
IR (KBr disk): 3274, 2917, 2857, 2095, 1746, 1719, 1233, 192, 1051 cm^-1
1H NMR (CDCl_Three, 400MHz) δ: 7.27 (m, 1H), 4.45 (dd, J = 8.30, 9.02 Hz, 1H), 4.41 (dd, J = 5.85, 9.02 Hz, 1H), 4.18 (ddd, J = 5.36, 5.61, 7.66 Hz 1H), 4.00 (dd, J = 5.85, 7.56 Hz, 1H), 3.89 (m, 1H), 3.26 (t, J = 6.84 Hz, 2H), 1.72-1.29 (m, 24H);
¹³C NMR (CDCl_Three, 100MHz) δ: 160.11, 108.37, 79.09, 76.82, 67.73, 51.67, 51.46, 29.38, 29.32, 29.09, 28.79, 27.76, 26.68, 25.25.
[0029]
Example 6
Synthesis of N-Boc body (27)
To a solution of the azide (26) (738 mg, 2.00 mmol) in N, N-dimethylformamide (10.0 mL) at 0 ° C., 4-N, N-dimethylaminopyridine (122 mg, 1.00 mmol), triethylamine (0.45 mL, 3.00 mmol) and di-t-butyl dicarbonate (655 mg, 3.00 mmol) were sequentially added and stirred for 3 hours. A saturated aqueous ammonium chloride solution was added to the reaction mixture for neutralization, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography (from 11% to 20% ethyl acetate in hexane) to obtain N-Boc body (27) (925 mg, 98.5%).
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[≡]_D ^24.0≡63.48 (c = 1.042, CHCl_Three)
IR (Neat): 3437, 2932, 2857, 2097, 1823, 1798, 1721, 1333, 1260, 1209, 1161, 1080 cm^-1
1H NMR (CDCl_Three, 400MHz) δ: 4.53 (dd, J = 7.32, 7.32 Hz, 1H), 4.45 (dd, J = 6.34, 6.34 Hz, 1H), 4.26 (m 3H), 3.26 (t, J = 6.83 Hz, 2H) , 1.65-1.29 (m, 33H);
¹³C NMR (CDCl_Three, 100MHz) δ: 152.06, 149.60, 108.57, 84.03, 76.28, 75.40, 62.57, 55.69, 51.44, 29.73, 29.46, 29.37, 29.33, 29.08, 28.79, 28.01, 26.83, 26.66, 26.08, 24.50.
[0030]
Example 7
Synthesis of alcohol (28)
Cesium carbonate (322 mg, 0.987 mmol) was added to a solution of N-Boc body (27) (925 mg, 1.97 mmol) in methanol (0.99 mL), and the mixture was stirred at room temperature for 1.5 hours. The reaction mixture was neutralized with a saturated aqueous ammonium chloride solution, concentrated under reduced pressure, and extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography (from 20% to 25% ethyl acetate in hexane) to obtain an alcohol form (28) (820 mg, 93.8%).
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[≡]_D ^24.09.83 (c = 0.543, CHCl_Three)
IR (Neat): 3360, 2928, 2930, 2857, 2097, 1698, 1524, 1368, 1248, 1171, 1046 cm^-1
1H NMR (CDCl_Three, 400MHz) δ: 4.96 (d, J = 8.44 Hz, 1H), 4.17 (m, 1H), 4.08 (dd, J = 6.10, 6.10 Hz, 1H), 3.84 (m 1H), 3.77 (m, 1H) , 3.69 (ddd, J = 3.66, 7.07, 10.73, 1H), 3.25 (t, J = 6.83 Hz, 2H), 2.33 (br, 1H), 1.67-1.28 (m, 33H);
¹³C NMR (CDCl_Three, 100MHz) δ: 155.39, 108.07, 79.73, 78.17, 77.83, 63.80, 51.47, 51.06, 29.51, 29.41, 29.20, 29.12, 28.81, 28.33, 27.60, 26.68, 25.26.
[0031]
Example 8
Synthesis of amine body (29)
Water (1.8 mL) and triphenylphosphine (706 mg, 2.69 mmol) were added sequentially to a solution of the alcohol (28) (794 mg, 1.79 mmol) in tetrahydrofuran (16.1 mL), then the temperature was raised to 60 ° C. for 3 hours. Stir. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography (from 13% to 25% methanoal in chloroform) to obtain an amine compound (29) (709 mg, 94.9%).
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[≡]_D ^24.08.34 (c = 0.873, CHCl_Three)
IR (KBr disk): 3362, 2928, 1686, 1528, 1368, 1173 cm^-1
1H NMR (CDCl_Three, 400MHz) δ: 7.80 (d, J = 7.80 Hz, 1H), 4.16 (m, 1H), 4.08 (dd, J = 6.10, 6.10 Hz, 1H), 3.83 (dd, J = 2.69, 10.74, 1H) , 3.76 (m, 1H), 3.69 (dd, J = 3.17, 10.97, 1H), 2.69 (t, J = 6.83 Hz, 2H), 2.12 (brs, 3H), 1.56-1.28 (m, 33H);
¹³C NMR (CDCl_Three, 100MHz) δ: 155.39, 108.00, 77.93, 77.84, 63.52, 51.09, 42.01, 33.40, 29.50, 29.44, 29.39, 29.14, 28.33, 27.64, 26.80, 26.67, 25.32.
[0032]
Example 9
Synthesis of NBD body (30)
To a solution of the amine compound (29) (648 mg, 1.56 mmol) in tetrahydrofuran (7.80 mL) at 0 ° C., triethylamine (0.35 mL, 2.33 mmol), 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole ( 466 mg, 2.33 mmol) was sequentially added, and the mixture was warmed to room temperature and stirred for 1 hour. A saturated aqueous ammonium chloride solution was added to the reaction mixture for neutralization, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography (from 25% to 50% ethyl acetate in hexane) to obtain NBD form (30) (666 mg, 73.9%).
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[≡]_D ^25.02.70 (c = 0.888, CHCl_Three)
IR (KBr disk): 3343, 2930, 1698, 1586, 1304, 1169, 1044 cm^-1
1H NMR (CDCl_Three, 400MHz) δ: 8.49 (d, J = 8.78 Hz, 1H), 6.44 (brs, 1H), 6.18 (d, J = 8.78 Hz, 1H), 4.98 (d, J = 9.27 Hz, 1H), 4.17 ( m, 1H), 4.09 (m, 1H), 3.84 (m, 1H), 3.78 (m, 1H), 3.70 (ddd, J = 3.66, 7.07, 10.98, 1H), 3.50 (t, J = 7.07 Hz, 1H), 3.49 (t, J = 7.07 Hz, 1H), 2.39 (brs, 1H), 1.81 (ddd, J = 7.32, 7.32, 14.64, 2H) 1.57-1.29 (m, 35H);
¹³C NMR (CDCl_Three, 100MHz) δ: 155.45, 144.24, 144.13, 143.94, 136.53, 108.08, 98.47, 79.74, 78.19, 77.81, 63.78, 51.09, 43.99, 29.46, 29.31, 29.20, 29.09, 28.45, 28.32, 27.57, 26.87, 26.66, 25.25 .
[0033]
Example 10
Synthesis of deacetonide (31)
To a solution of NBD body (30) (578 mg, 1.00 mmol) in methanol (5.00 mL) was added 2N aqueous hydrochloric acid solution (2.00 mL) at 0 ° C., then the mixture was warmed to room temperature and stirred for 3.5 hours. Saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture for neutralization, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated saline solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography (from 50% to 75% ethyl acetate in hexane) to obtain a deacetonide compound (31) (446 mg, 82.9%).
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[≡]_D ^25.06.11 (c = 0892, CHCl_Three)
IR (KBr disk): 3339, 2928, 1688, 1588, 1304, 1171cm^-1
1H NMR (CDCl_Three, 400MHz) δ: 8.48 (d, J = 8.79 Hz, 1H), 6.69 (brs, 1H), 6.18 (d, J = 8.79 Hz, 1H), 5.43 (d, J = 8.30 Hz, 1H), 3.89- 3.65 (m, 5H), 3.51 (t, J = 6.35 Hz, 1H), 3.50 (t, J = 6.59, 1H), 3.31 (brs, 1H), 1.94 (brs, 2H), 1.81 (m, 2H) , 1.68-1.28 (m, 25H);
¹³C NMR (CDCl_Three, 100MHz) δ: 156.46, 144.25, 144.11, 143.93, 136.66, 123.52, 98.53, 80.10, 76.18, 72.89, 61.90, 52.90, 44.03, 32.98, 29.39, 29.32, 29.23, 29.21, 29.03, 28.34, 26.82, 25.73.
[0034]
Example 11
Synthesis of NBD-phytosphingosine (32)
Trimethylsilyl bromide (0.28 mL, 2.13 mmol) was added dropwise to a solution of the deacetonide compound (31) (230 mg, 0.426 mmol) in methanol (2.1 mL) at room temperature, and the mixture was stirred for 1.5 hours. The reaction mixture was concentrated under reduced pressure, and the residue was separated and purified by reverse phase HPLC to obtain NBD-phytosphingosine (32) (171 mg, 88.6%).
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[≡]_D ^25.0≡1.79 (c = 0.699, CH_ThreeOH)
IR (KBr disk): 3341, 2926, 2855, 1588, 1298, 1269 cm^-1
1H NMR (CD_ThreeOD, 400MHz) δ: 8.49 (d, J = 8.78 Hz, 1H), 6.33 (d, J = 9.03 Hz, 1H), 3.94 (dd, J = 3.42, 11.47 Hz, 1H), 3.77 (dd, J = 8.54, 11.47 Hz, 1H), 3.57-3.44 (m, 5H), 1.81-1.32 (m, 18H);
¹³C NMR (CD_Three(OD, 100MHz) δ: 146.66, 145.79, 145.53, 138.56, 122.77, 99.59, 73.37, 73.36, 58.82, 56.45, 44.84, 35.38, 30.77, 30.69, 30.60, 30.34, 29.28, 28.04, 26.23.
[0035]
Example 12
Synthesis of tetraacetyl derivative (33)
Acetic anhydride (0.22 mL) was added at 0 ° C. to a solution of NBD-phytosphingosine (32) (49 mg, 0.112 mmol) in pyridine (0.56 mL), and the mixture was warmed to room temperature and stirred for 3 hours. A saturated aqueous copper sulfate solution was added, followed by extraction with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography (from 25% to 20% ethyl acetate in hexane) to obtain a tetraacetyl form (33) (50 mg, 91.2% for 2steps).
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[≡]_D ^24.517.33 (c = 0.745, CHCl_Three)
IR (Neat): 3322, 2930, 1742, 1586, 1302, 1252, 1046 cm^-1
1H NMR (CDCl_Three, 400MHz) δ: 8.49 (d, J = 8.54 Hz, 1H), 6.54 (t, J = 4.88 Hz, 1H), 6.18 (d, J = 8.54 Hz, 1H), 6.03 (d, J = 9.52 Hz, 1H), 5.11 (dd, J = 3.17, 8.05 Hz 1H), 4.95 (dt, J = 3.17, 9.76 Hz, 1H), 4.49 (m, 1H), 4.30 (dd, J = 4.88, 11.46 Hz 1H), 4.03 (dd, J = 3.17, 11.71 Hz 1H), 3.51 (t, J = 6.83 Hz 1H), 3.49 (t, J = 7.07 Hz 1H), 2.08 (s, 3H), 2.05 (s, 6H), 2.02 (s, 3H) 1.20-1.85 (m, 18H);
¹³C NMR (CDCl_Three, 100MHz) δ: 98.8, 67.6, 63.0, 62.3, 32.8, 30.7, 29.7, 29.5, 29.45, 29.40, 29.3, 26.2, 25.7, 25.5, 19.6.

Claims (4)

Formula (1)

(In the formula, X means an alkyl group having 1 to 15 carbon atoms, and P ¹ means a hydroxyl-protecting group.)
By epoxidizing the carbon-carbon double bond of the compound represented by formula (2)

(Wherein X and P ¹ are the same as above)
A compound represented by formula (3) is obtained by regioselectively opening the epoxide.

(Wherein X and P ¹ are the same as above)
A compound represented by formula (4) is obtained by deprotecting the hydroxyl group.

(Wherein X is the same as above)
And acetalizing two adjacent hydroxyl groups to obtain a compound represented by formula (5)

(In the formula, X is the same as above. R ¹ and R ² are the same or different and each represents a hydrogen atom, an alkyl group having 1 to 4 carbon atoms, or a phenyl group, and R ¹ and R ² are adjacent carbon atoms. And may form a cycloalkyl ring having 3 to 6 carbon atoms.)
And the hydroxyl group is changed to azide to obtain a compound represented by formula (6)

(In the formula, X, R ¹ and R ² are the same as above.)
The compound represented by formula (7) is obtained by protecting the amino group.

(In the formula, X, R ¹ and R ² are the same as above. P ² means an amino protecting group.)
The compound represented by formula (8) is obtained by hydrolyzing the oxazolidinone ring and opening the ring.

(In the formula, X, R ¹ , R ² and P ² are the same as above.)
To obtain a compound represented by formula (9)

(In the formula, X, R ¹ , R ² and P ² are the same as above.)
A compound represented by formula (10), (11) or (12), which is reacted with a reagent having a fluorescent labeling group, and then, if necessary, a protecting group is eliminated.

(In the formula, Z means a fluorescent labeling group. X, R ¹ , R ² and P ² are the same as above.)

(In the formula, X, Z and P ² are the same as above.)

(Wherein X and Z are the same as above)
A method for producing a phytosphingosine analog represented by the formula: or a compound in which a hydroxyl group or an amino group thereof is protected. Formula (1)

(In the formula, X means an alkyl group having 1 to 15 carbon atoms, and P ¹ means a hydroxyl-protecting group.)
By epoxidizing the carbon-carbon double bond of the compound represented by formula (2)

(Wherein X and P ¹ are the same as above)
A compound represented by formula (3) is obtained by regioselectively opening the epoxide.

(Wherein X and P ¹ are the same as above)
A method for producing an intermediate of a phytosphingosine analog, characterized by obtaining a compound represented by the formula: Formula (1a)

(In the formula, X means an alkyl group having 1 to 15 carbon atoms, and P ¹ means a hydroxyl-protecting group.)
By performing asymmetric epoxidation on the optically active compound represented by formula (2a)

(Wherein X and P ¹ are the same as above)
To obtain a compound represented by formula (3a):

(Wherein X and P ¹ are the same as above)
A process for producing an intermediate of an optically active phytosphingosine analog, characterized in that a compound represented by the formula:

(In the formula, X represents an alkyl group having 1 to 15 carbon atoms, R ¹ and R ² are the same or different and each represents a hydrogen atom, an alkyl group having 1 to 4 carbon atoms, or a phenyl group; ¹ and R ² may form a cycloalkyl ring having 3 to 6 carbon atoms with adjacent carbon atoms.
In the compound represented by formula (6), the hydroxyl group is changed to azide.

(In the formula, X, R ¹ and R ² are the same as above.)
The compound represented by formula (7) is obtained by protecting the amino group.

(Wherein X, R ¹ and R ² are the same as above, P ² means an amino protecting group), and the compound represented by formula (8) is obtained by hydrolysis.

(Wherein X, R ¹ , R ² and P ² are the same as above) to obtain the compound represented by formula (9)

(In the formula, X, R ¹ , R ² and P ² are the same as above.)
A compound represented by formula (10), (11) or (12), which is reacted with a reagent having a fluorescent labeling group, and then, if necessary, a protecting group is eliminated.

(In the formula, Z means a fluorescent labeling group. X, R ¹ , R ² and P ² are the same as above.)

(In the formula, X, Z and P ² are the same as above.)

(Wherein X and Z are the same as above)
A method for producing a phytosphingosine analog represented by the formula: or a compound in which a hydroxyl group or an amino group thereof is protected.