JP3761958B2 - 3'-carbamoylalkyl nucleoside derivatives - Google Patents

Description

【０００７】
〔式中、Ｂはシトシン、ウラシル、４−Ｎ−メチルシトシン又は４−Ｎ，Ｎ−ジメチルシトシンの核酸塩基を示し、Ｚは一般式−ＣＨ₂ＣＯＮＲ⁴Ｒ⁵ （Ｒ⁴ 及びＲ⁵ は同一又は相異なって、水素原子、炭素数１〜６の直鎖状又は分枝状アルキル基又はベンジル基を示す）で表される基を示し、Ｒ¹ 、Ｒ² 及びＲ³ は同一又は相異なって、水素原子、後記の一般式（２）で表される置換シリル基又は生体内で容易に脱離しうるエステル形成残基を示す〕で表される３′−カルバモイルアルキルヌクレオシド誘導体又はその薬学的に許容される塩を提供するものである。
【０００８】
上記一般式（１）で表わされる本発明化合物は、優れた抗腫瘍活性を有し、種々の腫瘍の治療剤等の医薬として有効である。
従って、また、本発明は、上記一般式（１）の化合物又はその薬学的に許容される塩と、薬学的担体とを含有する医薬組成物を提供するものである。
更に、本発明は、上記一般式（１）の化合物又はその薬学的に許容される塩を有効成分とする医薬、特に抗腫瘍剤を提供するものである。
【０００９】
【発明の実施の形態】
上記一般式（１）中、Ｂで示される核酸残基は、シトシン、ウラシル、４−Ｎ−メチルシトシン又は４−Ｎ，Ｎ−ジメチルシトシンである。
【００１５】
また、Ｒ¹ 、Ｒ² 及びＲ³ で示される置換シリル基は下記一般式（２）
【００１６】
【化４】

【００１７】
（式中、Ｒ^a 、Ｒ^b 及びＲ^c は同一又は相異なって、低級アルキル基又はフェニル基を示す）で表される基であって、具体的には、例えばトリメチルシリル、トリエチルシリル、トリプロピルシリル、トリイソプロピルシリル、トリｔｅｒｔ−ブチルシリル、トリヘキシルシリル、ジメチルエチルシリル、ジメチルイソプロピルシリル、ジエチルイソプロピルシリル、ジイソプロピルメチルシリル、ジｔｅｒｔ−ブチルメチルシリル、ｔｅｒｔ−ブチルジメチルシリル基等の炭素数１〜６の直鎖状又は分枝状のアルキル基でトリ置換されたシリル基、ジフェニルメチルシリル、ジメチルフェニルシリル、ｔｅｒｔ−ブチルジフェニルシリル、トリフェニルシリル基等が挙げられる。
【００１８】
Ｒ¹ 、Ｒ² 及びＲ³ で示される生体内で容易に脱離しうるエステル形成残基とは、ヒトを含む哺乳動物の血液及び組織内で容易に開裂して対応する水酸基化合物（すなわち、Ｒ¹ 、Ｒ² 及び／又はＲ³ が水素原子となる化合物）を放出する無毒のエステル残基を意味し、該エステル形成残基としては、通常よく知られるヌクレオシドの水酸基を保護しエステルを形成するものであればよく、例えば置換基を有してもよい脂肪族アシル基又は置換基を有してもよい芳香族アシル基等のアシル基、低級アルキルカルバモイル基、アミノ酸残基等が挙げられる。
【００１９】
置換基を有していてもよい脂肪族又は芳香族のアシル基としては、例えば低級アルカノイル基、アリールカルボニル基、ヘテロ環カルボニル基、アリールオキシカルボニル基、低級アルコキシカルボニル基、アシルオキシアシル基等が挙げられる。
低級アルカノイル基としては、例えばホルミル、アセチル、プロピオニル、ブチリル、イソブチリル、ペンタノイル、ヘキサノイル、クロロアセチル、ジクロロアセチル、トリクロロアセチル、トリフルオロアセチル、メトキシアセチル、エトキシアセチル基等の置換基としてハロゲン原子、低級アルコキシ基等を有することのある炭素数１〜６のアルカノイル基が挙げられる。
【００２０】
アリールカルボニル基としては、例えばベンゾイル、α−ナフチルカルボニル、β−ナフチルカルボニル、２−メチルベンゾイル、３−メチルベンゾイル、４−メチルベンゾイル、２，４−ジメチルベンゾイル、４−エチルベンゾイル、２−メトキシベンゾイル、３−メトキシベンゾイル、４−メトキシベンゾイル、２，４−ジメトキシベンゾイル、４−エトキシベンゾイル、２−メトキシ−４−エトキシベンゾイル、４−プロポキシベンゾイル、２−クロロベンゾイル、３−クロロベンゾイル、４−クロロベンゾイル、２，３−ジクロロベンゾイル、２−ブロモベンゾイル、４−フルオロベンゾイル、２−カルボキシベンゾイル、３−カルボキシベンゾイル、４−カルボキシベンゾイル、２−シアノベンゾイル、４−シアノベンゾイル、２−ニトロベンゾイル、４−ニトロベンゾイル又は２，４−ジニトロベンゾイル基等の置換基として低級アルキル基、低級アルコキシ基、ハロゲン原子、カルボキシル基、ニトロ基及びシアノ基等を有することのあるベンゾイル又はナフチルカルボニル基が挙げられる。
【００２１】
ヘテロ環カルボニル基としては、例えば２−フラニルカルボニル、４−チアゾリルカルボニル、２−キノリルカルボニル、２−ピラジニルカルボニル、２−ピリジルカルボニル、３−ピリジルカルボニル、４−ピリジルカルボニル基等が挙げられる。
アリールオキシカルボニル基としては、例えばフェノキシカルボニル、α−ナフチルオキシカルボニル、β−ナフチルオキシカルボニル、２−メチルフェノキシカルボニル、３−メチルフェノキシカルボニル、４−メチルフェノキシカルボニル、２，４−ジメチルフェノキシカルボニル、４−エチルフェノキシカルボニル、２−メトキシフェノキシカルボニル、３−メトキシフェノキシカルボニル、４−メトキシフェノキシカルボニル、２，４−ジメトキシフェノキシカルボニル、４−エトキシフェノキシカルボニル、２−メトキシ−４−エトキシフェノキシカルボニル、２−クロロフェノキシカルボニル、３−クロロフェノキシカルボニル、４−クロロフェノキシカルボニル、２，３−ジクロロフェノキシカルボニル、２−ブロモフェノキシカルボニル、４−フルオロフェノキシカルボニル、β−メチル−α−ナフチルオキシカルボニル、β−クロロ−α−ナフチルオキシカルボニル基等が挙げられる。
【００２２】
低級アルコキシカルボニル基としては、例えばメトキシカルボニル、エトキシカルボニル、ｎ−プロポキシカルボニル、イソプロポキシカルボニル、ｎ−ブトキシカルボニル、イソブトキシカルボニル、ｓｅｃ−ブトキシカルボニル、ｔｅｒｔ−ブトキシカルボニル、ペンチルオキシカルボニル、ヘキシルオキシカルボニル基等の炭素数２〜７の直鎖状又は分枝状のアルコキシカルボニル基が挙げられる。
アシルオキシアシル基としては、例えばアセチルオキシアセチル、プロピオニルオキシアセチル、α−（アセチルオキシ）プロピオニル、β−（プロピオニルオキシ）プロピオニル基等が挙げられる。
【００２３】
低級アルキルカルバモイル基としては、例えばメチルカルバモイル、エチルカルバモイル、プロピルカルバモイル、ブチルカルバモイル、ペンチルカルバモイル、ヘキシルカルバモイル、ジメチルカルバモイル、ジエチルカルバモイル基等の炭素数１〜６の直鎖状又は分枝状のアルキル基でモノ又はジ置換されたカルバモイル基が挙げられる。
アミノ酸残基としては、アミノ酸のカルボキシル基から水酸基を除いて形成される基を示し、天然又は合成のアミノ酸のいずれの由来のものでもよく、該アミノ酸としては、例えばグリシン、アラニン、β−アラニン、バリン、イソロイシン等が挙げられるが、特開平１−１０４０９３号記載のアミノ酸残基であればいずれであってもよい。
【００２４】
その他エステル形成残基としては、例えばＴＨＥＯＤＯＲＡ Ｗ．ＧＲＥＥＮＥ，”ＰＲＯＴＥＣＴＩＶＥ ＧＲＯＵＰＳ ＩＮ ＯＲＧＡＮＩＣ ＳＹＮＴＨＥＳＩＳ Ｓｅｃｏｎｄ Ｅｄｉｔｉｏｎ”，ＪＯＨＮ ＷＩＬＥＹ ＆ ＳＯＮＳ，ＩＮＣ．（１９９１）、日本化学会編＜新実験化学講座４＞「有機化合物の合成と反応（Ｖ）」１１章 ｐ２４９５ 丸善（１９８３）、特開昭６１−１０６５９３号、特開昭６２−１４９６９６号、特開平１−１５３６９６号に記載の通常のエステル形成残基として慣用されるもののいずれであってもよい。
【００２６】
Ｚとしては、好適にはカルバモイルメチル、メチルカルバモイルメチル、エチルカルバモイルメチル、ベンジルカルバモイルメチル又はジメチルカルバモイルメチル基が挙げられ、更に好適にはカルバモイルメチル基が挙げられる。
Ｒ¹ 、Ｒ² 及びＲ³ で示される生体内で容易に脱離しうるエステル形成残基としては、好適にはアシル基が挙げられ、更に好適にはアセチル基、ベンゾイル基が挙げられる。
Ｒ¹ 、Ｒ² 及びＲ³ としては、好適には水素原子が挙げられる。
【００２７】
本発明の好ましい化合物は、Ｂがシトシン、ウラシル、４―Ｎ−メチルシトシン又は４−Ｎ，Ｎ−ジメチルシトシンであり、Ｚがカルバモイルメチル、メチルカルバモイルメチル、エチルカルバモイルメチル、ベンジルカルバモイルメチル又はジメチルカルバモイルメチル基であり、Ｒ¹、Ｒ²及びＲ³が水素原子である３'− カルバモイルアルキルヌクレオシド誘導体である。
【００２８】
また更に好ましい化合物は、一般式（１）において、Ｂがシトシン、ウラシル、４−Ｎ−メチルシトシン又は４−Ｎ，Ｎ−ジメチルシトシンであり、Ｚがカルバモイルメチル基であり、Ｒ¹ 、Ｒ² 及びＲ³ が水素原子である３′−カルバモイルアルキルヌクレオシド誘導体である。
【００２９】
本発明化合物は塩の形態をも包含するものであり、かかる塩としては薬学的に許容される塩であれば特に制限はなく、例えばＲ³ が水素原子であるものの場合には塩酸塩、臭化水素酸塩、硫酸塩等の無機酸塩、メタンスルホン酸塩、ベンゼンスルホン酸塩等の有機スルホン酸塩、酢酸塩、プロピオン酸塩、トリフルオロ酢酸塩等の脂肪族カルボン酸塩等の有機酸塩等の酸付加塩が例示される。また、本発明化合物はその水和物をも包含するものである。
【００３０】
一般式（１）で表される本発明化合物は、例えば下記反応工程式１〜３に従い製造することができる。
【００３１】
【化５】

【００３２】
（式中、Ｂ、Ｒ⁴ 及びＲ⁵ は前記に同じ。Ｒ¹′は置換シリル基を示す。）
置換シリル基としては、前記の一般式（２）で表される基及びテトライソプロピルジシロキシル（ＴＩＰＤＳ）基等が挙げられる。好ましくはトリメチルシリル、ｔｅｒｔ−ブチルジメチルシリル、メチルジイソプロピルシリル及びトリイソプロピルシリル基挙げられる。
【００３３】
（Ａ工程）
一般式（３）で表される化合物をテトラヒドロフラン中、ヨウ化サマリウムと反応させることにより一般式（４）で表されるラクトン体化合物を得る。
一般式（３）で表される化合物は、公知の化合物であるかもしくは公知の方法により得られるが、具体的には後記反応工程式４に従い製造することができる。反応の割合は一般式（３）の化合物に対し、ヨウ化サマリウムを１〜１０倍モル量、好ましくは１〜５倍モル量使用するのがよい。反応温度は最初−７８℃で混合し、次に０〜５０℃、好ましくは室温付近で行われ、反応時間は０．１〜５０時間、好ましくは１〜２４時間で反応は有利に進行する。
【００３４】
（Ｂ工程）
Ａ工程で得られた一般式（４）で表される化合物を、適当な溶媒中、Ｒ⁴Ｒ⁵ＮＨで表されるアミン化合物と反応させることにより、一般式（１−ａ）で表される本発明化合物を得る。
溶媒としては反応に関与しないものであれば特に限定されず、いずれのものでもよく、例えばメタノール、エタノール等の低級アルコール類、ジメチルホルムアミド等が挙げられる。
またこの反応の際、更にジメチルアミノピリジン、トリエチルアミン等のアミン類を触媒として添加してもよい。
【００３５】
反応の割合は一般式（４）で表される化合物に対し、アミン化合物を１〜１０倍モル量、触媒を使用する時は触媒量を使用するのが好ましい。反応温度はメタノール性アンモニアを使用した場合、−７０℃付近であり、置換アミン化合物を使用した場合は室温付近である。反応時間は０．１〜５０時間、好ましくは１〜２４時間で反応は有利に進行する。
【００３６】
（Ｃ工程）
一般式（１−ａ）で表される化合物を、低級アルコール中、アンモニウムフルオリドと反応させることにより、一般式（１−ｂ）で表される本発明化合物を得る。
反応の割合は一般式（１−ａ）で表される化合物に対し、アンモニウムフルオリドを５〜５０倍モル量、好ましくは１０〜３０倍モル量使用するのがよい。反応は加熱還流で行い、反応時間は０．１〜２０時間、好ましくは１〜１０時間で反応は有利に進行する。
【００３７】
【化６】

【００３８】
（式中、Ｂ及びＲ¹′は前記に同じ。Ｒ²′及びＲ³′は置換シリル基を示す。）Ｒ²′及びＲ³′で示されるシリル基としては、前記のＲ¹′と同様のものが挙げられる。
【００３９】
（Ｄ工程）
反応工程式１のＢ工程で得られた一般式（１−ａ）で表される化合物中、Ｒ⁴ 及びＲ⁵ が水素原子である化合物（１−ａ′）を、ジメチルホルムアミド中、イミダゾールの存在下、ｔｅｒｔ−ブチルジメチルシリルクロリドと反応させることにより、一般式（１−ｃ）で表される本発明化合物を得る。
反応の割合は一般式（１−ａ′）で表される化合物に対し、ｔｅｒｔ−ブチルジメチルシリルクロリドを１〜１０倍モル量、好ましくは１〜５倍モル量、イミダゾールを１〜１０倍モル量、好ましくは２〜７倍モル量使用するのがよい。反応温度は０〜１００℃、好ましくは室温付近であり、反応時間は０．１〜５０時間、好ましくは０．５〜２４時間で反応は有利に進行する。
【００４０】
（Ｅ工程）
Ｄ工程で得られた一般式（１−ｃ）で表される化合物を、ピリジン中、ｐ−トルエンスルホニルクロリドと反応させることにより、一般式（１−ｄ）で表される本発明化合物を得る。
反応の割合は一般式（１−ｃ）で表される化合物に対し、ｐ−トルエンスルホニルクロリドを１〜１０倍モル量、好ましくは１〜５倍モル量使用するのがよい。反応は加熱還流で行い、反応時間は０．１〜５０時間、好ましくは０．５〜２４時間で反応は有利に進行する。
【００４１】
（Ｆ工程）
反応工程式１のＣ工程と同様に反応させることにより、一般式（１−ｅ）で表される本発明化合物を得る。
【００４２】
【化７】

【００４３】
（式中、Ｚ、Ｒ¹′及びＲ³′は前記に同じ。Ｒ⁶ 及びＲ⁷ は同一又は相異なって、水素原子、低級アルキル基又は置換基を有してもよいベンジル基を示す。）
【００４４】
（Ｇ工程）
反応工程式２で得られた一般式（１−ｃ）及び（１−ｄ）で表される化合物中、Ｂがウラシルである化合物（１−ｆ）を、アセトニトリル中、２，４，６−トリイソプロピルベンゼンスルホニル、ジメチルアミノピリジン及びトリエチルアミンの存在下、Ｒ⁶Ｒ⁷ＮＨで表されるアミン水溶液と反応させることにより、一般式（１−ｇ）で表されるシトシン誘導体である本発明化合物に変換することができる。
【００４５】
反応の割合は一般式（１−ｆ）で表される化合物に対し、アミン化合物を１〜１０倍モル量、好ましくは１〜５倍モル量、２，４，６−トリイソプロピルベンゼンスルホニル、ジメチルアミノピリジン及びトリエチルアミンをそれぞれ１〜１０倍モル量、好ましくは１〜５倍モル量使用するのがよい。反応温度は０〜１００℃、好ましくは室温付近であり、反応時間は０．１〜１００時間、好ましくは１〜５０時間で反応は有利に進行する。
【００４６】
反応工程式１〜３で得られた一般式（１−ｂ）及び（１−ｅ）の化合物の２′位、３′位及び５′位水酸基へのエステル形成残基の導入を、従来公知の方法、例えば上記で挙げた”ＰＲＯＴＥＣＴＩＶＥ ＧＲＯＵＰＳ ＩＮ ＯＲＧＡＮＩＣ ＳＹＮＴＨＥＳＩＳ Ｓｅｃｏｎｄ Ｅｄｉｔｉｏｎ”や日本化学会編＜新実験化学講座４＞「有機化合物の合成と反応（Ｖ）」に従うか、又は特開昭５８−１５２８９８号、特開昭６０−５６９９６号、特開昭６１−１０６５９３号、特開昭６２−１４９６９６号、特開平１−１５３６９６号等に記載の方法に従い行うことにより、他の本発明化合物へと導くことができる。
【００４７】
また、上記反応により得られた本発明化合物は、これを適当な溶媒中、前記無機酸又は有機酸と反応させる等の従来公知の方法により、塩の形態とすることができる。溶媒としては、例えば水、メタノール、エタノール、ジクロロメタン、テトラヒドロフラン、酢酸エチル、ヘキサン等が例示できる。反応温度は０〜５０℃で行われるのが好ましい。
また、前記反応工程式１で用いる原料化合物（３）は、例えば下記の反応工程式４に従って製造される。
【００４８】
【化８】

【００４９】
（式中、Ｂ、Ｒ¹′及びＲ³′は前記に同じ。）
【００５０】
（Ｈ工程）
一般式（５）で表される化合物を部分加水分解、例えばＪ．Ｏｒｇ．Ｃｈｅｍ．，５５、４１０−４１２（１９９０）に記載の方法、すなわちトリフルオロ酢酸−水混合液中、０℃で反応させることにより、５′位の選択的な脱シリル化反応を行ない一般式（６）で表される化合物を得る。
一般式（５）で表される化合物は、公知の化合物であるかもしくは公知の方法、例えば上記のＪ．Ｏｒｇ．Ｃｈｅｍ．，ＳＹＮＴＨＥＳＩＳ、２８３−２８８（１９９１）、Ｔｅｔｒａｈｅｄｒｏｎ ４７、１７２７−１７３６（１９９１）に記載の方法により得られる。
【００５１】
（Ｉ工程）
Ｈ工程で得られた一般式（６）で表される化合物を適当な溶媒中、ブロモアセチルブロミドと反応させることにより、一般式（３）で表される化合物を得る。溶媒としては反応に関与しないものであれば特に限定されず、いずれのものでもよく、例えばクロロホルム、ジクロロメタン等のハロゲン化炭化水素類、ジメチルホルムアミド等が挙げられる。
またこの反応の際、更にジメチルアミノピリジン、トリエチルアミン、２，６−ルチジン等のアミン類を触媒として添加してもよい。
【００５２】
反応の割合は一般式（６）で表される化合物に対し、ブロモアセチルブロミドを１〜１０倍モル量、好ましくは１〜５倍モル量、触媒を使用する時は触媒量を使用するのが好ましい。反応温度は−７８℃付近がよい。反応時間は１分〜２４時間、好ましくは１分〜１時間で反応は有利に進行する。
【００５３】
上記方法により得られる本発明化合物及び各化合物は、通常公知の分離精製手段、例えば濃縮、溶媒抽出、濾過、再結晶、各種クロマトグラフィー等を用いることにより単離精製することができる。
【００５４】
本発明の化合物は、適当な薬学的担体を用いて通常の方法に従い、医薬組成物とすることができる。ここで用いられる担体としては、通常の薬剤に汎用される各種のもの、例えば賦形剤、結合剤、崩壊剤、滑沢剤、着色剤、矯味剤、矯臭剤、界面活性剤等を使用することができる。
【００５５】
本発明医薬又は医薬組成物をヒトを含む哺乳動物の腫瘍の治療剤として使用する際の投与単位形態は特に限定されず、治療目的に応じて適宜選択でき、具体的には注射剤、坐剤、外用剤（軟膏剤、貼付剤等）、エアゾール剤等の非経口剤、錠剤、被覆錠剤、散剤、顆粒剤、カプセル剤、丸剤、液剤（懸濁剤、乳剤等）等の経口剤が挙げられる。
上記各種組成物は、この分野で通常知られた製剤化方法により製剤化される。
【００５６】
注射剤の形態にするに際しては、担体として、例えば水、エチルアルコール、マクロゴール、プロピレングリコール、エトキシ化イソステアリルアルコール、ポリオキシ化イソステアリルアルコール、ポリオキシエチレンソルビタン脂肪酸エステル類等の希釈剤、クエン酸ナトリウム、酢酸ナトリウム、リン酸ナトリウム等のｐＨ調整剤及び緩衝剤、ピロ亜硫酸ナトリウム、エチレンジアミン四酢酸、チオグリコール酸、チオ乳酸等の安定化剤などを使用できる。尚、この場合、等張性の溶液を調製するに充分な量の食塩、ブドウ糖或いはグリセリンを医薬製剤中に含有せしめてもよく、また通常の溶解補助剤、無痛化剤、局所麻酔剤等を添加してもよい。これらの担体を添加して、常法により、皮下、筋肉内、静脈内用注射剤を製造することができる。
【００５７】
坐剤の形態にするに際しては、担体として、例えばポリエチレングリコール、カカオ脂、ラノリン、高級アルコール、高級アルコールのエステル類、ゼラチン、半合成グリセライド、ウィテップゾール（登録商標：ダイナマイトノーベル社）等に適当な吸収促進剤を添加して使用できる。
【００５８】
軟膏剤、例えばペースト、クリーム及びゲルの形態に調製する際には、通常使用される基剤、安定剤、湿潤剤、保存剤等が必要に応じて配合され、常法により混合、製剤化される。基剤としては、例えば白色ワセリン、パラフィン、グリセリン、セルロース誘導体、ポリエチレングリコール、シリコン、ベントナイト等を使用できる。保存剤としては、パラオキシ安息香酸メチル、パラオキシ安息香酸エチル、パラオキシ安息香酸プロピル等を使用できる。
【００５９】
貼付剤を製造する場合には、通常の支持体に上記軟膏、クリーム、ゲル、ペースト等を常法により塗布すればよい。支持体としては、綿、スフ、化学繊維からなる織布、不織布や軟質塩化ビニル、ポリエチレン、ポリウレタン等のフィルムあるいは発泡体シートが適当である。
【００６０】
錠剤、散剤、顆粒剤等の経口用固形製剤の形態にするに際しては、担体として、例えば乳糖、白糖、塩化ナトリウム、ブドウ糖、尿素、デンプン、炭酸カルシウム、カオリン、結晶セルロース、ケイ酸、メチルセルロース、グリセリン、アルギン酸ナトリウム、アラビアゴム等の賦形剤、単シロップ、ブドウ糖液、デンプン液、ゼラチン溶液、ポリビニルアルコール、ポリビニルエーテル、ポリビニルピロリドン、カルボキシメチルセルロース、セラック、メチルセルロース、エチルセルロース、水、エタノール、リン酸カリウム等の結合剤、乾燥デンプン、アルギン酸ナトリウム、カンテン末、ラミナラン末、炭酸水素ナトリウム、炭酸カルシウム、ポリオキシエチレンソルビタン脂肪酸エステル類、ラウリル硫酸ナトリウム、ステアリン酸モノグリセリド、デンプン、乳糖等の崩壊剤、白糖、ステアリン酸、カカオバター、水素添加油等の崩壊抑制剤、第４級アンモニウム塩基、ラウリル硫酸ナトリウム等の吸収促進剤、グリセリン、デンプン等の保湿剤、デンプン、乳糖、カオリン、ベントナイト、コロイド状ケイ酸等の吸着剤、精製タルク、ステアリン酸塩、ホウ酸末、ポリエチレングリコール等の滑沢剤等を使用できる。更に錠剤は、必要に応じ通常の剤皮を施した錠剤、例えば糖衣錠、ゼラチン被包錠、腸溶被錠、フィルムコーティング錠、二重錠、多層錠等とすることができる。
【００６１】
カプセル剤は上記で例示した各種の担体と混合し、硬質ゼラチンカプセル、軟質カプセル等に充填して調製される。
丸剤の形態にするに際しては、担体として、例えばブドウ糖、乳糖、デンプン、カカオ脂、硬化植物油、カオリン、タルク等の賦形剤、アラビアゴム末、トラガント末、ゼラチン、エタノール等の結合剤、ラミナラン、カンテン等の崩壊剤などを使用できる。
液体製剤は水性又は油性の懸濁液、溶液、シロップ、エリキシル剤であってもよく、これらは通常の添加剤を用いて常法に従い、調製される。
【００６２】
上記製剤中に含有されるべき本発明化合物の量は、剤型、投与経路、投与計画等により異なり一概には言えず、広い範囲から適宜選択されるが、通常製剤中に１〜７０重量％程度とするのが好ましい。
【００６３】
上記製剤の投与方法は特に限定されず、製剤の形態、患者等の投与対象の年齢、性別その他の条件、症状の程度等に応じて、例えば経腸投与、経口投与、直腸投与、口腔内投与、経皮投与等が適宜決定される。例えば錠剤、丸剤、液剤、懸濁剤、乳剤、顆粒剤及びカプセル剤の場合には経口投与され、坐剤の場合には直腸内投与される。注射剤の場合には単独で又はブドウ糖、アミノ酸等の通常の補液と混合して静脈内投与され、更に必要に応じて単独で動脈内、筋肉内、皮内、皮下もしくは腹腔内投与される。軟膏剤は、皮膚、口腔内粘膜等に塗布される。
【００６４】
本発明の化合物の投与量は、用法、患者等の投与対象の年齢、性別、状態、腫瘍の種類、投与される本発明化合物の種類、その他の条件等に応じて適宜選択されるが、一般に投与単位形態当たり経口剤では約１〜１０００ｍｇ、注射剤では約０．１〜５００ｍｇ、坐剤では約５〜１０００ｍｇとするのが望ましい。また、上記投与形態を有する薬剤の１日当たりの投与量は、通常０．１〜２００ｍｇ／ｋｇ体重／日程度、好ましくは０．５〜１００ｍｇ／ｋｇ体重／日程度の範囲となる量を目安とするのがよい。これら本発明製剤は１日に１回又は２〜４回程度に分けて投与することができる。
【００６５】
本発明化合物を含有する製剤を投与することにより治療できる悪性腫瘍としては、特に制限はなく、例えば頭頚部癌、食道癌、胃癌、結腸癌、直腸癌、肝臓癌、胆のう・胆管癌、膵臓癌、肺癌、乳癌、卵巣癌、膀胱癌、前立腺癌、睾丸腫瘍、骨・軟部肉腫、悪性リンパ腫、白血病、子宮頚癌、皮膚癌、脳腫瘍等が挙げられる。
【００６６】
【発明の効果】
本発明の３′−カルバモイルアルキルヌクレオシド誘導体は優れた抗腫瘍作用を有し、種々の腫瘍の治療剤等の医薬として有用である。
【００６７】
【実施例】
以下に参考例、実施例及び薬理試験例を示し、本発明を更に詳しく説明するが、本発明は何らこれらによって限定されるものではない。
【００６８】
参考例１
1-(5-O-ブロモアセチル-2-O-tert-ブチルジメチルシリル-β-D-エリスロ-ペントフラン-3-ウロシル)ウラシルの合成：
1-(2-O-tert-ブチルジメチルシリル-β-D-エリスロ-ペントフラン-3-ウロシル)ウラシル 4.48 g (12.6 mmol) をジクロロメタン 30 mlに溶解し、アルゴン雰囲気下、-78℃に冷却後、2,6-ルチジン 2.20ml(17.6 mmol)及び臭化ブロモアセチル1.41ml(15.1mmol)を滴下し、５分間撹拌した。反応混合液に飽和炭酸水素ナトリウム水溶液10mlを加え、ジクロロメタン層を抽出し、水 (３×30ml）、飽和食塩水（30ml）で洗浄後、無水硫酸ナトリウムで乾燥した。濾過後、濾液を減圧下留去し、残渣を少量の酢酸エチルに溶解し、シリカゲルに吸着させ溶媒を留去した後、シリカゲルカラムクロマトグラフィー (４×20cm，30％酢酸エチル/ヘキサン) により精製し、標記化合物4.12g （収率70％）を黄色泡状物質として得た。
【００６９】
FAB-MS : m/z 478 (MH⁺)
¹H-NMR (CDCl₃) δ： 9.67 (br s, 1H, NH-3, exchanged with D₂O),
7.49(d, 1H, H-6, J_6,5 = 8.2Hz), 5.97 (d,1H, H-1', J_1',_2'= 7.1Hz),
5.84 (d, 1H, H-5, J_5,6 = 8.2Hz),
4.51 (d, 1H, H-5'a, J_5'a,5'b = 11.9Hz),
4.47 (d, 1H, H-2', J_2', _1' = 7.1Hz),
4.44 (dd, 1H, H-4', J_4',_5'a = 2.0, J_4' ,_5'b = 3.5Hz),
4.41 (dd, 1H, H-5b', J_5'b,4' = 3.5, J_5'b,_5'a = 11.9Hz),
0.83 (s, 9H, t-Bu), 0.09, 0.02 (各 s, 各 3H, Me) .
¹³C-NMR (CDCl₃) δ： 206.2, 166.4, 163.2, 150.6, 140.3, 104.1,
87.93, 78.77, 75.35, 64.54, 25.81, 25.64,
25.54, 18.33, -4.4, -5.01.
【００７０】
参考例２
一般式（４）で表されるラクトン体（Ｂはウラシル、Ｒ¹′はtert-ブチルジメチルシリル）の合成：
0.1Nヨウ化サマリウム-テトラヒドロフラン溶液14.2ml(1.24mmol)をアルゴン雰囲気下、-78℃に冷却後、参考例１で得た化合物338mg(0.567mmol)をテトラヒドロフラン6mlに溶解したものを、ゆっくり滴下した。反応混合液を室温に戻し、1N塩酸4mlを加えて撹拌した後、酢酸エチル50mlで抽出し、水 (２×20ml) 、飽和食塩水 (20ml) で洗浄後、無水硫酸ナトリウムで乾燥した。濾過後、濾液を減圧下留去し、残渣を少量の酢酸エチルに溶解し、シリカゲルに吸着させ溶媒を留去した後、シリカゲルカラムクロマトグラフィー (２×11cm, 30％酢酸エチル/ヘキサン) により精製し、標記のラクトン体219mg(収率75％)を黄白色グラス状物質として得た。分析用試料として酢酸エチル-ヘキサンより結晶化し淡黄色結晶を得た。
【００７１】
FAB-MS : m/z 399 (MH⁺)
¹H-NMR (CDCl₃) δ： 8.19 (br s, 1H, NH-3, exchanged with D₂O),
7.16 (d, 1H, H-6, J_6,5 = 8.1Hz),
5.98 (d,1H, H-1', J_1',_2' = 7.5Hz),
5.84 (d, 1H, H-5, J_5,6 = 8.1Hz),
4.46 (d, 1H, H-5'a, J_5'a,_5'b = 13.1Hz), 4.41 (br s, 1H, H-4'),
4.37 (dd, 1H, H-5'b, J_5'b, _4' = 2.4, J_5'b,_5'a = 13.1Hz),
3.66 (d, 1H, H-2', J_2',_1' = 7.5Hz),
3.49 (s, 1H, 3'-OH, exchanged with D₂O),
2.78, 2.71( 各 d, 各 1H, H-3''a, H-3''b, J = 15.3Hz),
0.91 (s, 9H, t-Bu), 0.11, -0.02 (各 s, 各 3H, Me).
¹³C-NMR (CDCl₃) δ： 169.6, 162.4, 150.3, 131.6, 104.4, 85.02,
81.39, 74.08, 68.36, 40.63, 25.68, 17.97, 11.68, -4.52, -4.64.
【００７２】
実施例１
1-(2-O-tert-ブチルジメチルシリル-3-C-カルバモイルメチル-β-D-リボフラノシル)ウラシル（化合物１）の合成：
参考例２で得たラクトン化合物808mg(2.03mmol)をメタノール5mlに溶解し、-7０℃に冷却したアンモニア飽和メタノール溶液20mlを加え、-70℃で3時間放置した。反応混合液を減圧下留去し、残渣を少量のメタノールに溶解し、シリカゲルに吸着させ溶媒を留去した後、シリカゲルカラムクロマトグラフィー (４×10cm)により精製し、標記化合物１を825mg(収率98％)を白色粉状物質として得た。分析用試料としてメタノール/クロロホルムより結晶化し淡黄色針状結晶を得た。
【００７３】
FAB-MS : m/z 415 (MH⁺)
元素分析：
計算値（C₁₇H₂₉N₃O₇Siとして）：C, 49.26; H, 6.81; N, 10.14.
実測値：C, 49.10; H, 7.02; N, 10.13.
【００７４】
実施例２
1-(2,5-O-ジ-tert-ブチルジメチルシリル-3-C-カルバモイルメチル-β-D-リボフラノシル)ウラシル（化合物２）の合成：
実施例１で得た化合物１の1.50g(3.62mmol)をジメチルホルムアミド50mlに溶解し、イミダゾール738mg(10.7mmol)及びtert-ブチルジメチルシリルクロリド819mg(5.43mmol)を加え、室温で１２時間撹拌した。反応混合液にエタノール5mlを加えて15分間撹拌し、溶媒を減圧下留去した。残渣に酢酸エチル100ml及び水100mlを加えて分配し、さらに酢酸エチル層を水(３×50ml)、飽和食塩水(50ml)で洗浄後、無水硫酸ナトリウムで乾燥した。濾過後、濾液を減圧下留去し、残渣をシリカゲルカラムクロマトグラフィー (４×8.5cm,４％メタノール/クロロホルム)により精製し、標記化合物２を1.9g(収率99％)黄色泡状物質として得た。
【００７５】
FAB-MS : m/z 529 (MH⁺)
¹H-NMR (CDCl₃) δ： 8.14 (br s, 1H, NH-3, exchanged with D₂O),
8.00 (d, 1H, H-6, J_6,5 = 8.2Hz),
6.88 (br s, 1H, アミド, exchanged with D₂O),
6.17 (d, 1H, H-1', J_1',_2' = 7.2Hz),
5.74 (dd, 1H, H-5, J_5,6 = 8.2Hz, J₅,_NH-3 = 2.0Hz),
5.42 (br s, 1H, アミド, exchanged with D₂O),
4.27 (br s, 1H, H-4'), 4.07 (d, 1H, H-2', J_2',_1' = 7.2Hz),
3.91 (dd, 1H, H-5'a, J_5'a,_4' = 2.1, J_5'a,_5'b = 12.1Hz),
3.87 (dd, 1H, H-5'b, J_5'b,_4' = 2.2, J_5'b,_5'a = 12.1Hz),
3.47 (s, 1H, 3'-OH, exchanged with D₂O),
2.64, 2.56 (各 d, 各 1H, H-3''a, H-3''b, J = 15.9Hz),
0.97, 0.91 (各 s, 各 9H, t-Bu),
0.23, 0.22, 0.06, -0.05 (各 s, 各 3H, Me).
¹³C-NMR (CDCl₃) δ： 172.3, 163.19, 151.0, 140.54, 103.49, 86.12,
85.10, 78.84, 78.31, 63.31, 40.68, 26.24, 25.80, 18.54, 18.00,
11.69, -4.35, -4.50, -5.10, -5.52.
【００７６】
実施例３
1-(2,5-O-ジ-tert-ブチルジメチルシリル-3-C-カルバモイルメチル-β-D-リボフラノシル)シトシン（化合物３）の合成：
実施例２で得た化合物２の1.05g(2mmol)をアルゴン雰囲気下、アセトニトリル50mlに溶解し、塩化2,4,6-トリイソプロピルベンゼンスルホニル1.21g(4.00mmol)、ジメチルアミノピリジン487mg(4.00mmol)及びトリエチルアミン0.28ml(4.00mmol)を加え室温で24時間撹拌後、濃アンモニア水 (28％,20ml)を加えて更に室温で2 時間撹拌した。反応混合液を減圧下溶媒留去し、残渣を少量のメタノールに溶解し、シリカゲルに吸着させ溶媒を留去した後、シリカゲルカラムクロマトグラフィー (２×18cm, 20％メタノール/クロロホルム) により精製し、標記化合物３を830mg(収率76％)白色結晶として得た。分析用試料としてメタノール/クロロホルムより再結晶し、白色平板状結晶を得た。
【００７７】
FAB-MS : m/z 530 (MH⁺)
¹H-NMR (CDCl₃) δ： 8.04 (d, 1H, H-6, J_6,5 = 8.2Hz),
7.23 (br s, 1H, アミド a, exchanged with D₂O),
6.31 (d, 1H, H-1', J_1',_2' =7.2Hz), 5.95 (br s, 1H, H-5),
5.49 (br s, 1H, アミド b, exchanged with D₂O),
4.24 (br s, 1H, H-4'), 4.06 (d, 1H, H-2', J_2',1' = 7.2Hz),
4.01 (d, 1H, H-5'a, J_5'a,5'b = 11.8Hz),
3.89 (dd, 1H, H-5'b, J_5'b,_4' = 2.0, J_5'b,5'a = 12.2Hz),
2.59 (t, 2H, H-3''a, H-3''b, J = 15.1Hz),
0.96, 0.85 ( 各 s, 各 9H, t-Bu),
0.17, -0.01, -0.10 (各 s, 各 3H, Me).
【００８２】
実施例６
1-(3-C-カルバモイルメチル-β-D-リボフラノシル)ウラシル（化合物６）の合成：
実施例１で得た化合物１の281mg(0.678mmol)をメタノール20mlに溶解し、アンモニウムフロリド502mg(13.6mmol)を加え、2時間加熱還流した。反応混合液を減圧下溶媒留去し、残渣に水50ml及びクロロホルム50mlを加えて分配し、さらに水層をクロロホルム (２×30ml)で洗浄後、減圧下20mlまで濃縮し、上清にUV吸収がなくなるまで活性炭を加え、カラムにつめて水100mlで洗浄し、50％メタノール水溶液で溶出した。溶出液を減圧下濃縮し、標記化合物６を205mg(収率99％)白色粉状物質として得た。分析用試料として水/メタノールより結晶化し、白色針状結晶を得た。
【００８３】
FAB-MS : m/z 302 (MH⁺)
¹H-NMR (DMSO-d₆) δ： 9.45 (br s, 1H, NH-3, exchanged with D₂O),
8.05 (d, 1H, H-6, J_6,5 = 8.0Hz),
7.47, 7.06 (各 br s, 各 1H, アミド, exchanged with D₂O),
5.91 (d, 1H, H-1', J_1', _2' = 7.9Hz),
5.67 (d, 1H, H-5, J_5,6 = 8.1Hz),
5.56 (d, 1H, 2'-OH, J_2'-OH,2' = 6.1Hz, exchanged with D₂O),
5.23 (t, 1H, 5'-OH, J _5'-OH,5' = 3.9Hz, exchanged with D₂O),
5.19 (s, 1H, 3'-OH, exchanged with D₂O), 3.96 (br s, 1H, H-4'),
3.94 (dd, 1H, H-2', J_2',2'-OH = 6.3, J_2',1' = 7.8Hz),
3.71 (ddd, 1H, H-5'a, J_5'a,4' = 1.5,
J_5'a,5'b = 12.0, J _5'a,5'-OH = 3.4Hz),
3.59 (ddd, 1H, H-5'b, J_5'b,4' = 3.0,
J_5'b,5'a = 12.0, J_5'b,5'-OH = 3.4Hz),
2.57, 2.50 (各 d, 各 1H, H-3''a, 3''b, J = 15.3Hz).
【００８４】
実施例７
1-(3-C-カルバモイルメチル-β-D-リボフラノシル)シトシン (化合物７) の合成：
実施例３で得た化合物３の105mg(0.2mmol)をメタノール5mlに溶解し、アンモニウムフロリド148mg(4.00mmol)を加え、2時間加熱還流した。反応混合液を減圧下溶媒留去し、残渣に水10ml及びクロロホルム10mlを加えて分配し、さらに水層をクロロホルム (２×10ml)で洗浄後、減圧下5mlまで濃縮し、上清にUV吸収がなくなるまで活性炭を加え、カラムにつめて水50mlで洗浄し、50％メタノール水溶液で溶出した。溶出液を減圧下濃縮し、標記化合物７を47.5mg(収率79％)白色粉状物質として得た。分析用試料として水/メタノールより結晶化し白色粉末を得た。
【００８５】
FAB-MS : m/z 301 (MH⁺)
¹H-NMR (DMSO-d₆) δ： 7.90 (d, 1H, H-6, J_6,5 = 7.4Hz),
7.47 (br s, 1H, アミド a, exchanged with D₂O),
7.17 (br s, 2H, NH₂, exchanged with D₂O),
7.05 (br s, 1H, アミド b, exchanged with D₂O),
5.88 (d, 1H, H-1', J_1',2' = 7.9Hz), 5.75 (br s, 1H, H-5) _,
5.41 (d, 1H, 2'-OH, J_2'-OH,2' = 6.2Hz, exchanged with D₂O),
5.22 (t, 1H, 5'-OH, J_5'-OH,5' = 4.5Hz, exchanged with D₂O),
5.13 (s, 1H, 3'-OH, exchanged with D₂O),
3.98 (dd, 1H, H-2', J_2',2'-OH = 7.1, J_2',1' = 7.1Hz),
3.93 (br s, 1H, H-4'),
3.66 (ddd, 1H, H-5'a,
J_5'a,4' = 2.9, J _5'a,5'b = 9.8, J_5'a,5'-OH = 4.5Hz),
3.57 (ddd, 1H, H-5'b, J_5'b,4' = 2.4,
J_5'b,5'a = 9.7, J_5'b,5'-OH = 4.5Hz),
2.55, 2.49 (各 d, 各 1H, H-3''a, 3''b, J = 16.3Hz).
【００９０】
実施例１０
1-[2-O-tert-ブチルジメチルシリル-3-C-(N-メチルカルバモイルメチル)-β-D-リボフラノシル]ウラシル（化合物１０）の合成：
参考例２で得たラクトン化合物398mg(1mmol)をジメチルホルムアミド5mlに溶解し、メチルアミン塩酸塩333mg(5mmol)及びトリエチルアミン700 μl(5.00mmol)を加え室温で7時間撹拌した。反応混合液を減圧下留去し、残渣を酢酸エチル100ml及び水50mlで分配し、さらに酢酸エチル層を水 (３×50ml)、飽和食塩水(50ml)で洗浄後、無水硫酸ナトリウムで乾燥した。濾過後、濾液を減圧下留去し、残渣をシリカゲルカラムクロマトグラフィー (２×６cm,４％メタノール/クロロホルム) により精製し、標記化合物１０を456mg(収率99％)黄色グラス状物質として得た。
【００９１】
FAB-MS : m/z 430 (MH⁺)
¹H-NMR (CDCl₃) δ： 8.15 (br s, 1H, NH-3, exchanged with D₂O),
7.70 (d, 1H, H-6, J_6,5 = 8.2Hz),
6.60 (d, 1H,アミド, J _NH,Me = 4.7Hz, exchanged with D₂O),
5.79 (dd, 1H, H-5, J_5,6 = 8.2, J_5,NH-3 = 2.2Hz),
6.17 (d, 1H, H-1', J_1',2' = 7.3Hz),
4.07 (d, 1H, H-2', J_2',1' = 7.3Hz),
4.27 (br s, 1H, H-4'),
3.88 (dd, 1H, H-5'a, J_5'a,4' = 2.4, J_5'a,5'b = 12.6Hz),
3.76 (dd, 1H, H-5'b, J_5'b,4' = 1.2, J_5'b,5'a = 12.6Hz),
3.54 (s, 1H, 3'-OH, exchanged with D₂O),
2.83 (d, 3H, Me, J_Me,NH = 4.7Hz),
2.80, 2.56 (各 d, 各 1H, H-3''a, H-3''b, J = 15.7Hz),
0.89 (s, 9H, t-Bu), 0.63, -0.01 (各 s, 各 3H, Me).
【００９２】
実施例１１
1-[2-O-tert-ブチルジメチルシリル-3-C-(N,N-ジメチルカルバモイルメチル)-β-D-リボフラノシル]ウラシル（化合物１１）の合成：
参考例２で得たラクトン化合物398mg(1mmol)から実施例１０と同様にして、黄色グラス状物質として標記化合物１１を収率99％で得た。
【００９３】
FAB-MS : m/z 444 (MH⁺)
¹H-NMR (CDCl₃)δ： 8.19 (br s, 1H, NH-3, exchanged with D₂O),
7.32 (d 1H, H-6, J_6,5 = 8.0Hz),
5.77 (dd, 1H, H-5, J_5,6 = 8.0, J_5,NH-3 = 2.2Hz),
5.41 (d, 1H, H-1', J_1',2' = 7.7Hz),
4.75 (d, 1H, H-2', J_2',1' = 7.7Hz),
4.39 (br s, 1H, H-4'),
3.83 (dd, 1H, H-5'a, J_5'a,4' = 2.3, J_5'a,5'b = 13.1Hz),
3.57 (dd, 1H, H-5'b, J_5'b,4' = 1.0, J_5'b,5'a = 1.3Hz),
3.03, 2.97 (各 s, 各 3H, Me),
2.99, 2.75 (各 d, 各 1H, H-3''a, H-3''b, J = 16.5Hz),
0.89 (s, 9H, t-Bu), 0.86, -0.06 (各 s, 各 3H, Me) .
【００９４】
実施例１２
1-[2-O-tert-ブチルジメチルシリル-3-C-(N-エチルカルバモイルメチル)-β-D-リボフラノシル]ウラシル（化合物１２）の合成：
参考例２で得たラクトン化合物398mg(1mmol)から実施例１０と同様にして、黄色グラス状物質として標記化合物１２を収率99％で得た。
【００９５】
FAB-MS : m/z 444 (MH⁺)
¹H-NMR (CDCl₃) δ： 9.28 (br s, 1H, NH-3, exchanged with D₂O),
7.78 (d, 1H, H-6, J_6,5 = 8.1Hz),
6.80 (d, 1H,アミド, J_NH,CH2 = 5.3Hz, exchanged with D₂O),
5.79 (d, 1H, H-1', J_1',2' = 7.3Hz),
5.71 (dd, 1H, H-5, J_5,6 = 8.1, J_5,NH-3 = 2.2Hz),
4.40 (d, 1H, H-2', J_2',1' = 7.3Hz),
4.20 (br s, 1H, 5'-OH, exchanged with D₂O),
4.17 (s, 1H, 3'-OH, exchanged with D₂O),
4.01 (br s, 1H, H-4'),
3.80 (br d, 1H, H-5'a, J_5'a,4' = 1.0, J_5'a,5'b = 12.1Hz),
3.73 (dd, 1H, H-5'b, J_5'b,4' = 5.0, J_5'b,5'a = 12.1Hz),
3.26 (dq, 2H, CH₂CH₃, J_CH2,NH = 5.3, J = 7.1Hz),
2.73, 2.51 (各 d, 各 1H, H-3''a, H-3''b, J = 15.7Hz),
1.12 (t, 1H, Me, J = 7.1Hz), 0.89 (s, 9H, t-Bu),
0.63, -0.01 (各 s, 各 3H, Me).
【００９６】
実施例１３
1-[2-O-tert-ブチルジメチルシリル-3-C-(N-ベンジルカルバモイルメチル)-β-D-リボフラノシル]ウラシル（化合物１３）の合成：
参考例２で得たラクトン化合物398mg(1mmol)から実施例１０と同様にして、無色グラス状物質として標記化合物１３を収率80％で得た。
【００９７】
FAB-MS : m/z 506 (MH⁺)
¹H-NMR (CDCl₃) δ： 8.10 (br s, 1H, NH-3, exchanged with D₂O),
8.05 (d, 1H, H-6, J_6,5 = 8.3Hz), 7.37-7.33 (m, 5H, aromatic),
7.15 (t, 1H,アミド, J_NH,CH = 5.6Hz, exchanged with D₂O),
6.15 (d, 1H, H-1', J_1',2' = 7.3Hz),
5.72 (dd, 1H, H-5, J_5,6 = 8.1, J_5,NH-3 = 2.3Hz),
4.56 (d, 2H, PhCH₂, J = 5.6Hz), 4.19 (br s, 1H, H-4'),
4.07 (d, 1H, H-2', J_2',1' = 7.3Hz),
3.87 (dd, 1H, H-5'a, J_5'a,4' = 1.2, J _5'a,5'b = 12.5Hz),
3.82 (dd, 1H, H-5'b, J_5'b,4' = 2.1, J_5'b,5'a = 12.5Hz),
3.40 (s, 1H, 3'-OH, exchanged with D₂O),
2.68, 2.44 (各 d, 各 1H, H-3''a, H-3''b, J = 15.5Hz),
0.96, 0.89 (各 s, 各 9H, t-Bu),
0.18, 0.17, 0.03, -0.07 (各 s, 各 3H, Me).
【００９８】
実施例１４
1-[2,5-O-ジ-tert-ブチルジメチルシリル-3-C-(N-メチルカルバモイルメチル)-β-D-リボフラノシル]ウラシル（化合物１４）の合成：
実施例１０で得た化合物１０の300mg(0.7mmol)をジメチルホルムアミド20mlに溶解し、イミダゾール190mg(2.8mmol)及びtert-ブチルジメチルシリルクロリド211mg(1.4mmol)を加え室温で１時間撹拌した。反応混合液にエタノール 2mlを加えて15分間撹拌し、溶媒を減圧下留去した。残渣に酢酸エチル50ml及び水20mlを加えて分配し、さらに酢酸エチル層を水 (３×20ml)、飽和食塩水(20ml)で洗浄後、無水硫酸ナトリウムで乾燥した。濾過後、濾液を減圧下留去し、残渣をシリカゲルカラムクロマトグラフィー (２×6cm,40％酢酸エチル/ヘキサン) により精製し、標記化合物１４を398mg(収率100％) 白色泡状物質として得た。
【００９９】
FAB-MS : m/z 544 (MH⁺)
¹H-NMR (CDCl₃) δ： 8.06 (br s, 1H, NH-3, exchanged with D₂O),
8.03 (d, 1H, H-6, J_6,5 = 8.2Hz),
6.79 (d, 1H, アミド, J_NH,Me = 4.7Hz, exchanged with D₂O),
6.16 (d , 1H, H-1', J_1',2' = 7.2Hz),
5.73 (dd, 1H, H-5, J_5,6 = 8.1, J_5,NH-3 = 2.2Hz),
4.22 (br s, 1H, H-4'), 4.06 (d, 1H, H-2', J_2',1' = 7.2Hz),
3.88 (br s, 2H, H-5'a,b),
3.52 (s, 1H, 3'-OH, exchanged with D₂O),
2.81 (d, 3H, Me, J_Me,NH = 4.7Hz),
2.63, 2.51 (各 d, 各 1H, H-3''a, H-3''b, J = 15.5Hz),
0.96, 0.89 (各 s, 各 9H, t-Bu),
0.18, 0.10, 0.03, -0.06 (各 s, 各 3H, Me).
【０１００】
実施例１５
1-[2,5-O-ジ-tert-ブチルジメチルシリル-3-C-(N,N-ジメチルカルバモイルメチル)-β-D-リボフラノシル]ウラシル（化合物１５）の合成：
実施例１１で得た化合物１１の192mg(0.44mmol)から実施例１４と同様にして、白色泡状物質として標記化合物１５を収率85％で得た。
【０１０１】
FAB-MS : m/z 558 (MH⁺)
¹H-NMR (CDCl₃) δ： 7.97 (br s, 1H, NH-3, exchanged with D₂O),
7 .88 (d, 1H, H-6, J_6,5 = 8.1Hz),
6.23 (d, 1H, H-1', J_1',2' = 7.5Hz),
5.71 (dd, 1H, H-5, J_5,6 = 8.1, J_5,NH-3 = 2.3Hz),
4.46 (br s, 1H, H-4'), 4.04 (d, 1H, H-2', J_2',1' = 7.4Hz),
3.88 (dd, 1H, H-5'a, J_5'a,4' = 1.2, J_5'a,5'b = 12.0Hz),
3.86 (dd, 1H, H-5'b, J_5'b,4' = 2.4, J_5'b,5'a = 12.0Hz),
3.51 (s, 1H, 3'-OH, exchanged with D₂O),
3.00, 2.98 (各 s, 各 3H, Me),
2.80, 2.71 (各 d, 各 1H, H-3''a, H-3''b, J = 16.3Hz),
0.95, 0.90 (各 s, 各 9H, t-Bu),
0.12, 0.11, 0.04, -0.07 (各 s, 各 3H, Me)_.
【０１０２】
実施例１６
1-[2,5-O-ジ-tert-ブチルジメチルシリル-3-C-(N-エチルカルバモイルメチル)-β-D-リボフラノシル]ウラシル（化合物１６）の合成：
実施例１２で得た化合物１２の222mg(0.5mmol)から実施例１４と同様にして、白色泡状物質として標記化合物１６を257mg(収率92％)得た。
【０１０３】
FAB-MS : m/z 558 (MH⁺)
¹H-NMR (CDCl₃) δ： 7.97 (br s, 1H, NH-3, exchanged with D₂O),
7.88 (d, 1H, H-6, J_6,5 = 8.1Hz),
6.23 (d, 1H, H-1', J_1',2' = 7.5Hz),
5.71 (dd, 1H, H-5, J_5,6 = 8.1, J_5,NH-3 = 2.3Hz),
4.46 (br s, 1H, H-4'), 4.04 (d, 1H, H-2', J_2',1' = 7.4Hz),
3.88 (dd, 1H, H-5'a, J_5'a,4' = 1.2, J_5'a,5'b = 12.0Hz),
3.86 (dd, 1H, H-5'b, J_5'b,4' = 2.4, J_5'b,5'a = 12.0Hz),
3.51 (s, 1H, 3'-OH, exchanged with D₂O),
3.00, 2.98 (各 s, 各 3H, Me),
2.80, 2.71 (各 d, 各 1H, H-3''a, H-3''b, J = 16.3Hz),
0.95, 0.90 (各 s, 各 9H, t-Bu),
0.12, 0.11, 0.04, -0.07 (各 s, 各 3H, Me).
【０１０４】
実施例１７
1-[2,5-O-ジ-tert-ブチルジメチルシリル-3-C-(N-ベンジルカルバモイルメチル)-β-D-リボフラノシル]ウラシル（化合物１７）の合成：
実施例１３で得た化合物１３の404mg(0.8mmol)から実施例１４と同様にして、白色泡状物質として標記化合物１７を460mg(収率93％) 得た。
【０１０５】
FAB-MS : m/z 620(MH⁺)
¹H-NMR (CDCl₃) δ： 8.06 (br s, 1H, NH-3, exchanged with D₂O),
8.01 (d, 1H, H-6, J_6,5 = 8.3Hz), 7.37-7.26 (m, 5H, aromatic),
7.13 (t, 1H,アミド, J_NH,CH = 5.7Hz, exchanged with D₂O),
6.15 (d, 1H, H-1', J_1',2' = 7.3Hz),
5.72 (dd, 1H, H-5, J_5,6 = 8.1, J_5,NH-3 = 2.3Hz),
4.46-4.45 (各 d, 各 1H, PhCH ₂, J = 5.8Hz),
4.19 (br s, 1H, H-4'), 4.07 (d, 1H, H-2', J_2',1' = 7.3Hz),
3.87 (dd, 1H, H-5'a, J_5'a,4' = 1.2, J_5'a,5'b = 12.5Hz),
3.82 (dd, 1H, H-5'b, J_5'b,4' = 2.1, J _5'b,5'a = 12.5Hz),
3.40 (s, 1H, 3'-OH, exchanged with D₂O),
2.68, 2.44 (各 d, 各 1H, H-3''a, H-3''b, J = 15.5Hz),
0.96, 0.89 (各 s, 各 9H, t-Bu),
0.18, 0.17, 0.03, -0.07 (各 s, 各 3H, Me).
【０１０６】
実施例１８
1-[2,5-O-ジ-tert-ブチルジメチルシリル-3-C-(N-メチルカルバモイルメチル)-β-D-リボフラノシル]シトシン（化合物１８）の合成：
実施例１４で得た化合物１４の217mg(0.4mmol)をアルゴン雰囲気下、アセトニトリル10mlに溶解し、塩化2,4,6-トリイソプロピルベンゼンスルホニル242mg(0.8mmol)、ジメチルアミノピリジン97mg(0.8mmol)及びトリエチルアミン0.056ml(0.8mmol)を加え室温で24時間撹拌後、濃アンモニア水 (28％_, 10ml)を加えて更に室温で２時間撹拌した。反応混合液を減圧下溶媒留去し、残渣を少量のメタノールに溶解し、シリカゲルに吸着させ溶媒を留去した後、シリカゲルカラムクロマトグラフィー (２×8cm, 10％メタノール/クロロホルム) により精製し、標記化合物１８を157mg(収率73％)白色結晶として得た。分析用試料としてメタノール/クロロホルムより再結晶し、白色平板状結晶を得た。
【０１０７】
FAB-MS : m/z 543 (MH⁺)
¹H-NMR (CDCl₃) δ： 8.06 (d, 1H, H-6, J_6,5 = 8.2Hz),
7.05 (d, 1H,アミド, J _NH,Me = 4.5Hz, exchanged with D₂O),
6.23 (d, 1H, H-1', J_1',2' = 6.7Hz), 6.15 (br s, 1H, H-5),
4.19 (br s, 1H, H-4'), 4.05 (d, 1H, H-2', J_2',1' = 6.7Hz),
3.99 (d, 1H, H-5'a, J_5'a,5'b = 11.5Hz),
3.87 (d, 1H, H-5'b, J_5'b,5'a = 11.5Hz),
2.80 (d, 3H, Me, J_Me,NH = 4.5Hz),
2.60, 2.53 (各 d, 各 1H, H-3''a, H-3''b, J = 15.1Hz),
0.95, 0.84 (各 s, 各 9H, t-Bu),
0.17, 0.16, -0.01, -0.03 (各 s, 各 3H, Me).
【０１０８】
実施例１９
1-[2,5-O-ジ-tert-ブチルジメチルシリル-3-C-(N,N-ジメチルカルバモイルメチル)-β-D-リボフラノシル]シトシン（化合物１９）の合成：
実施例１５で得た化合物１５の167mg(0.3mmol)から実施例１８と同様にして、白色粉状物質として標記化合物１９を115mg(収率69％) 得た。
【０１０９】
FAB-MS : m/z 557(MH⁺)
¹H-NMR (CDCl₃) δ： 8.15 (d, 1H, H-6, J_6,5 = 6.5Hz),
6.30 (br s, 1H, H-5), 6.23 (d, 1H, H-1', J_1',2' = 7.3Hz),
4.43 (br s, 1H, H-4'), 4.03 (d, 1H, H-2', J_2',1' = 7.3Hz),
3.87 (br s, 2H, H-5'a, b), 2.98, 2.96 (各 s, 各 3H, Me),
2.76, 2.74 (各 d, 各 1H, H-3''a, H-3''b, J = 16.8Hz),
0.93, 0.88 (各 s, 各 9H, t-Bu),
0.12, 0.10, 0.02, -0.01 ( 各 s, 各 3H, Me).
【０１１０】
実施例２０
1-[2,5-O-ジ-tert-ブチルジメチルシリル-3-C-(N-エチルカルバモイルメチル)-β-D-リボフラノシル]シトシン（化合物２０）の合成：
実施例１６で得た化合物１６の280mg(0.5mmol)から実施例１８と同様にして、無色グラス状物質として標記化合物２０を280mg(収率100％)得た。
【０１１１】
FAB-MS : m/z 557(MH⁺)
¹H-NMR (CDCl₃) δ： 8.02 (d, 1H, H-6, J_6,5 = 7.5Hz),
7.10 (br s, 1H, アミド, exchanged with D₂O),
6.31 (d, 1H, H-5, J_5,6 = 7.5Hz),
6.31 (d, 1H, H-1', J_1',2' = 7.1Hz),
4.15 (br s, 1H, H-4'), 4.05 (d, 1H, H-2', J_2',1' = 7.1Hz),
3.95 (d, 1H, H-5'a, J_5'a,5'b = 11.8Hz),
3.85 (dd, 1H, H-5'b, J_5'b,4' = 11.8Hz),
3.27 (dq, 2H, NCH₂ , J = 7.2Hz),
2.59, 2.52 (各 d, 各 1H, H-3''a, H-3''b, J = 16.8Hz),
1.13 (t, 3H, NCH₂CH₃, J = 7.2Hz),
0.95, 0.86 (各 s, 各 9H, t-Bu),
0.16, 0.15, -0.04, -0.10 (各 s, 各 3H, Me).
【０１１２】
実施例２１
1-[2,5-O-ジ-tert-ブチルジメチルシリル-3-C-(N-ベンジルカルバモイルメチル)-β-D-リボフラノシル]シトシン（化合物２１）の合成：
実施例１７で得た化合物１７の402mg(0.640mmol)から実施例１８と同様にして、白色粉状物質として標記化合物２１を352mg(収率89.0％)得た。
【０１１３】
FAB-MS : m/z 619 (MH⁺)
¹H-NMR (CDCl₃) δ： 8.03 (d, 1H, H-6, J_6,5 = 7.4Hz),
7.56 (br s, 1H, アミド, exchanged with D₂O),
7.34-7.24 (m, 5H, aromatic),
6.31 (d, 1H, H-1', J_1',2' = 7.3Hz),
5.88 (br s, 1H, H-5), 4.44 (s, 2H, PhCH₂),
4.15 (br s, 1H, H-4'), 4.06 (d, 1H, H-2', J_2',1' = 7.3Hz),
3.97 (d, 1H, H-5'a, J_5'a,5'b = 11.8Hz),
3.82 (dd, 1H, H-5'b, J_5'b,4' = 1.9, J _5'b,5'a = 11.8Hz),
2.65, 2.59 (各 d, 各 1H, H-3''a, H-3''b, J = 15.1Hz),
0.95, 0.82 (各 s, 各 9H, t-Bu),
0.16, 0.15, -0.04, -0.06 (各 s, 各 3H, Me)
【０１１４】
実施例２２
1-[3-C-(N-メチルカルバモイルメチル)-β-D-リボフラノシル]シトシン（化合物２２）の合成：
実施例１８で得た化合物１８の108mg(0.2mmol)をメタノール5mlに溶解し、アンモニウムフルオリド74mg(4mmol)を加え、２時間加熱還流した。反応混合液を減圧下溶媒留去し、残渣に水10ml及びクロロホルム10mlを加えて分配し、さらに水層をクロロホルム (２×10ml)で洗浄後、減圧下10mlまで濃縮し、上清にUV吸収がなくなるまで活性炭を加え、カラムにつめて水50mlで洗浄し、50％メタノール水溶液で溶出した。溶出液を減圧下濃縮し、標記化合物２２を54mg(収率86％)白色粉状物質として得た。分析用試料として水/メタノールより結晶化し白色粉状結晶を得た。
【０１１５】
FAB-MS : m/z 315 (MH⁺)
¹H-NMR (DMSO-d₆) δ： 8.05 (d, 1H, H-6, J_6,5 = 8.1Hz),
7.90 (q, 1H,アミド, J = 4.5Hz, exchanged with D₂O),
7.28 (d, 2H, 4-NH₂, exchanged with D₂O),
5.91 (d, 1H, H-1', J_1',2' = 8.0Hz),
5.68 (d, 1H, H-5, J_5,6 = 8.1Hz),
5.58 (d, 1H, 2'-OH, J_2'-OH,2' = 6.0Hz, exchanged with D₂O),
5.25 (t, 1H, 5'-OH, J_5'-OH,5' = 3.9Hz, exchanged with D₂O),
5.10 (s, 1H, 3'-OH, exchanged with D₂O),
3.96-3.93 (m, 2H, H-2',4'),
3.75 (ddd, 1H, H-5'a, J_5'a,4' = 1.5, J_5,a,5'b = 12.1, J_5'a,5'-OH
= 3.9Hz),
3.59 (ddd, 1H, H-5'b, J_5'b,4' = 3.5, J_5'b,5'a=
12.1, J _5'b,5'-OH = 3.9Hz),
2.55, 2.54 (各 d, 各 1H, H-3''a, 3''b, J = 16.2Hz).
【０１１６】
実施例２３
1-[3-C-(N, N-ジメチルカルバモイルメチル)-β-D-リボフラノシル]シトシン（化合物２３）の合成：
実施例１９で得た化合物１９の111mg(0.2mmol)から実施例２２と同様にして、白色粉状物質として標記化合物２３を60.5mg(収率92％)得た。
【０１１７】
FAB-MS : m/z 329 (MH⁺)
¹H-NMR (DMSO-d₆) δ：7.96 (d, 1H, H-6, J_6,5 = 7.3Hz),
7.26 (br d, 2H, 4-NH₂, exchanged with D₂O),
5.91 (d, 1H, H-1', J_1',2' = 8.1Hz),
5.75 (d, 1H, H-5, J_5,6 = 7.3Hz),
5.27 (t, 1H, 5'-OH, J_5'-OH,5' = 3.9Hz, exchanged with D₂O),
5.26 (d, 1H, 2'-OH, J_2'-OH,2' = 5.4Hz, exchanged with D₂O),
5.17 (s, 1H, 3'-OH, exchanged with D₂O),
4.03 (br s, 1H, H-4'),
3.97 (dd, 1H, H-2', J_2',2'-OH = 5.4, J_2',1' = 7.3Hz),
3.58 (br s, 2H, H-5'a,b), 2.99, 2.85 (各 d, 各 3H, Me),
2.80, 2.70 (各 d, 各 1H, H-3''a, 3''b, J = 15.3Hz)
【０１１８】
実施例２４
1-[3-C-(N-エチルカルバモイルメチル)-β-D-リボフラノシル]シトシン（化合物２４）の合成：
実施例２０で得た化合物２０の111mg(0.2mmol)から実施例２２と同様にして、白色粉状物質として標記化合物２４を56.9mg(収率86％)得た。
【０１１９】
FAB-MS : m/z 329 (MH⁺)
¹H-NMR (DMSO-d₆) δ： 8.32 (d, 1H, H-6, J_6,5 = 7.8Hz),
8.03 (t, 1H,アミド, J_NH,CH2 = 5.4Hz, exchanged with D₂O),
6.95 (br s, 2H, 4-NH₂, exchanged with D₂O),
6.08 (d, 1H, H-1', J_1',2' = 7.8Hz),
5.93 (d, 1H, H-5, J_5,6 = 7.8Hz),
5.76 (d, 1H, 2'-OH, J_2'-OH,2' = 6.5Hz, exchanged with D₂O),
5.33 (t, 1H, 5'-OH, J_5'-OH,5' = 3.2Hz, exchanged with D₂O),
5.21 (s, 1H, 3'-OH, exchanged with D₂O),
4.56 (dq, 2H, CH₂CH₃ , J_CH2,NH = 5.4, J = 7.3Hz),
4.02 (br s, 1H, H-4'),
3.96 (dd, 1H, H-2', J_2',2'-OH = 6.5, J_2',1' = 7.8Hz),
3.77 (ddd, 1H, H-5'a, J_5'a,4' = 1.3, J_5'a,5'b =
12.5, J_5'a,5'-OH = 3.2Hz),
3.61 (ddd, 1H, H-5'b, J_5'b,4' = 3.2, J _5'b,5'a =
12.5, J _5'b,5'-OH = 3.2Hz),
2.55, 2.54 (各 d, 各 1H, H-3''a, 3''b, J = 15.3Hz),
1.01 (t, 3H, CH₃ , J = 7.3Hz).
【０１２０】
実施例２５
1-[3-C-(N-ベンジルカルバモイルメチル)-β-D-リボフラノシル]シトシン（化合物２５）の合成：
実施例２１で得た化合物２１の124mg(0.2mmol)から実施例２２と同様にして、白色粉状物質として標記化合物２５を55.4mg(収率71％)得た。
【０１２１】
FAB-MS : m/z 391 (MH⁺)
¹H-NMR (DMSO-d₆) δ： 8.45 (br t, 1H,アミド, J_NH,CH2 = 5.8Hz,
exchanged with D₂O), 7.91 (d, 1H, H-6, J_6,5 = 7.4Hz),
7.33-7.22 (m, 5H, aromatic),
7.16 (br s, 2H, 4-NH₂ , exchanged with D₂O),
5.99 (d, 1H, H-1', J_1',2' = 7.9Hz),
5.78 (br s, 1H, H-5),
5.42 (d, 1H, 2'-OH, J_2'-OH,2'= 6.1Hz, exchanged with D₂O),
5.20 (t, 1H, 5'-OH, J_5'-OH,5' = 4.4Hz, exchanged with D₂O),
5.02 (s, 1H, 3'-OH, exchanged with D₂O),
4.30 (d, 2H, PhCH₂ , J_CH2,NH = 5.8Hz),
4.02 (dd, 1H, H-2', J_2',2'-OH = 6.1, J_2',1' = 7.9Hz),
3.96 (br s, 1H, H-4'),
3.70 (ddd, 1H, H-5'a, J_5'a,4' = 1.2, J_5'a,5'b =
11.9, J_5'a,5'-OH = 4.4Hz),
3.58 (ddd, 1H, H-5'b, J_5'b,4' = 3.1, J_5'b,5'a =
11.9, J_5'a,5'-OH = 4.4Hz),
2.65, 2.61 (各 d, 各 1H, H-3''a, 3''b, J = 15.1Hz)
【０１２２】
実施例２６
1-(2,5-O-ジ-tert-ブチルジメチルシリル-3-C-カルバモイルメチル-β-D-リボフラノシル)-4-N-メチルシトシン（化合物２６）の合成：
実施例２で得た化合物２の265mg(0.5mmol)をアルゴン雰囲気下、アセトニトリル20mlに溶解し、塩化2,4,6-トリイソプロピルベンゼンスルホニル450mg(1.5mmol)、ジメチルアミノピリジン182mg(1.5mmol)及びトリエチルアミン0.105ml(1.5mmol)を加え室温で24時間撹拌後、50％メチルアミン水溶液5mlを加え、更に室温で2時間撹拌した。反応混合液を減圧下溶媒留去し、残渣をクロロホルム50ml及び水20mlで分配し、クロロホルム層をさらに水 (２×20ml)、飽和食塩水(20ml)で洗浄後、無水硫酸ナトリウムで乾燥した。濾過後、濾液を減圧下留去し、残渣をシリカゲルカラムクロマトグラフィー (２×15cm, 5％メタノール/クロロホルム) により精製し、標記化合物２６を256mg(収率94％) 黄色結晶として得た。分析用試料としてクロロホルムより再結晶し、白色平板状結晶を得た。
【０１２３】
FAB-MS : m/z 543 (MH⁺)
¹H-NMR (CDCl₃) δ： 8.13 (br s, 1H,アミド a, exchanged with D₂O),
7.79 (d, 1H, H-6, J_6,5 = 8.1Hz),
7.05 (br s, 1H, 4-NH, exchanged with D₂O),
6.24 (d, 1H, H-1', J_1',2' = 7.2Hz), 5.71 (br s, 1H, H-5),
5.47 (br s, 1H, アミド b, exchanged with D₂O),
4.21 (br s, 1H, H-4'), 4.10 (d, 1H, H-2', J_2',1' = 7.2Hz),
3.86 (br s, 2H, H-5'a,b), 2.81 (d, 3H, Me),
2.60 (t, 2H, H-3''a, H-3''b, J = 15.8Hz),
0.93, 0.86 (各 s, 各 9H, t-Bu),
0.18, 0.17, -0.01, -0.10 (各 s, 各 3H, Me).
【０１２４】
実施例２７
1-(2,5-O-ジ-tert-ブチルジメチルシリル-3-C-カルバモイルメチル-β-D-リボフラノシル)-4-N,N-ジメチルシトシン（化合物２７）の合成：
実施例２で得た化合物２の167mg(0.3mmol)と50％ジメチルアミン水溶液5mlから実施例２６と同様にして、黄色グラス状物質として標記化合物２７を240mg(収率86％)得た。
【０１２５】
FAB-MS : m/z 543 (MH⁺)
¹H-NMR (CDCl₃) δ： 7.99 (d, 1H, H-6, J_6,5 = 7.9Hz),
7.10 (br s, 1H, アミド a, exchanged with D₂O),
6.27 (d, 1H, H-1', J_1',2' = 6.7Hz),
5.82 (d, 1H, H-5, J = 7.6Hz),
5.39 (br s, 1H, アミド b, exchanged with D₂O),
4.18 (br s, 1H, H-4'), 4.08 (d, 1H, H-2', J_2',1' = 6.7Hz),
3.38 (dd, 1H, H-5'a, J_5'a,4' = 2.9, J _5'a,5'b = 12.2Hz),
3.28, 3.12 (各 br s, 各 3H, Me),
2.62, 2.54 (各 d, 各 1H, H-3''a, H-3''b, J = 15.7Hz),
0.97, 0.86 (各 s, 各 9H, t-Bu),
0.17, 0.16, -0.00, -0.05 (各 s, 各 3H, Me).
【０１２６】
実施例２８
1-(3-C-カルバモイルメチル-β-D-リボフラノシル)-4-N-メチルシトシン（化合物２８）の合成：
実施例２６で得た化合物２６の108mg(0.2mmol)をメタノール5mlに溶解し、アンモニウムフルオリド74mg(4mmol)を加え、2時間加熱還流した。反応混合液を減圧下溶媒留去し、残渣に水10ml及びクロロホルム10mlを加えて分配し、さらに水層をクロロホルム (２×10ml)で洗浄後、減圧下10mlまで濃縮し、上清にUV吸収がなくなるまで活性炭を加え、カラムにつめて水50mlで洗浄し、50％メタノール水溶液で溶出した。溶出液を減圧下濃縮し、標記化合物２８を50mg(収率80％)白色粉状物質として得た。分析用試料として水/メタノールより結晶化し白色粉状結晶を得た。
【０１２７】
FAB-MS : m/z 315 (MH⁺)
¹H-NMR (DMSO-d₆) δ： 7.96 (d, 1H, H-6, J_6,5 = 7.4Hz),
7.24 (br d, 1H, 4-NH, exchanged with D₂O),
7.28 (d, 2H, アミド, exchanged with D₂O),
5.93 (d, 1H, H-5, J_5,6 = 7.9Hz),
5.78 (d, 1H, H-1', J_1',2' = 7.4Hz),
5.50 (d, 1H, 2'-OH, J_2'-OH,2' = 6.0Hz, exchanged with D₂O),
5.32 (t, 1H, 5'-OH, J _5'-OH,5' = 4.2Hz, exchanged with D₂O),
5.10 (s, 1H, 3'-OH, exchanged with D₂O),
4.03 (t, 2H, H-2', J_2',1' = 7.4, J_2',2'-OH = 4.2Hz),
3.97 (br s, 1H, H-4'),
3.74 (ddd, 1H, H-5'a, J _5'a,4' = 3.0, J_5'a,5'b =
11.9, J_5'a,5'-OH = 4.2Hz),
3.62 (ddd, 1H, H-5'b, J_5'b,4' = 3.2, J_5'b,5'a =
11.9, J_5'b,5'-OH = 4.2Hz), 2.63 (d, 3H, Me, J_Me,NH = 4.5Hz),
2.57 (s, 2H, H-3''a, 3''b)_.
【０１２８】
薬理試験例１（殺細胞作用）
ヒトＫＢ細胞を１×１０⁵ cells/wellで９６穴プレートに播種した。本発明化合物を精製水に溶解させた後、ＲＰＭＩ １６４０メディウムで種々の濃度に希釈し、各wellに添加して培養した。５％ＣＯ₂ インキュベーターで３７℃、３日間の接触後、ＭＴＴ法により細胞数を計測した。
各化合物の殺細胞作用を、コントロールの細胞数を５０％減少させる薬剤濃度（ＩＣ₅₀)として表した。結果を表１に示す。
【０１２９】
【表１】

【０１３０】
この結果から明らかなように、本発明化合物は、既知化合物である１−（３−Ｃ−エチル−β−Ｄ−リボフラノシル）ウラシルと比較して極めて強力な殺細胞活性を示した。
【０１３１】
薬理試験例２（殺細胞作用）
表２に示す癌細胞を使用し、薬理試験例１と同様にして、化合物７のＩＣ₅₀を求めた。結果を表２に示す。
【０１３２】
【表２】

【０１３３】
以下に本発明化合物を用いた製剤例を示す。
製剤例１（カプセル剤）
カプセル剤を下記配合割合で常法に従い調製した。
【表３】
化合物３ ２００mg
乳糖 ３０mg
トウモロコシデンプン ５０mg
結晶セルロース １０mg
ステアリン酸マグネシウム ３ mg
１カプセル当たり ２９３mg
【０１３４】
製剤例２（錠剤）
錠剤を下記配合割合で常法に従い調製した。
【表４】
化合物７ １００mg
乳糖 ４７mg
トウモロコシデンプン ５０mg
結晶セルロース ５０mg
ヒドロキシプロピルセルロース １５mg
タルク ２mg
ステアリン酸マグネシウム ２mg
エチルセルロース ３０mg
不飽和脂肪酸グリセリド ２mg
二酸化チタン ２ mg
１錠当たり ３００mg
【０１３５】
製剤例３（顆粒剤）
顆粒剤を下記配合割合で常法に従い調製した。
【表５】
化合物２４ ２００mg
マンニトール ５４０mg
トウモロコシデンプン １００mg
結晶セルロース １００mg
ヒドロキシプロピルセルロース ５０mg
タルク １０ mg
１包当たり １０００mg
【０１３６】
製剤例４（細粒剤）
細粒剤を下記配合割合で常法に従い調製した。
【表６】
化合物２８ ２００mg
マンニトール ５２０mg
トウモロコシデンプン １００mg
結晶セルロース １００mg
ヒドロキシプロピルセルロース ７０mg
タルク １０ mg
１包当たり １０００mg
【０１３７】
製剤例５（注射剤）
注射剤を下記配合割合で常法に従い調製した。
【表７】
化合物７ １００mg
注射用蒸留水 適量
１管中 ２ml
【０１３８】
製剤例６（坐剤）
坐剤を下記配合割合で常法に従い調製した。
【表８】
化合物６ ２００mg
ウイテップゾールＳ−５５
（ラウリン酸からステアリン酸までの飽和脂肪酸
のモノ−、ジ−、トリ−グリセライド混合物、
ダイナマイトノーベル社製） １３００ mg
１個あたり １５００mg[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a novel nucleic acid derivative, and more particularly to a 3'-carbamoylalkyl nucleoside derivative having excellent antitumor activity and useful as a medicament such as an antitumor agent.
[0002]
[Prior art]
Conventionally, as an antitumor agent which is a nucleic acid antimetabolite, pyrimidine compounds such as 5-fluorouracil, tegafur, UFT (DFT), doxyfluridine, carmofur, cytarabine, and inositabine are known.
On the other hand, pyrimidines or purine nucleosides having an alkyl group at the 3-position of the sugar moiety are described in JP-B Nos. 45-11908 and 46-4376, but their anti-tumor effects are extremely weak and are useful as anticancer agents. There is no utility value at present.
[0003]
[Problems to be solved by the invention]
Accordingly, an object of the present invention is to provide a novel nucleic acid derivative having excellent antitumor activity and useful as a medicament such as an antitumor agent.
[0004]
[Means for Solving the Problems]
In this situation, the present inventor has conducted extensive research and found that a specific nucleic acid derivative having a substituent introduced at the 3-position of sugar exhibits excellent antitumor activity and is useful as an antitumor agent. The present invention has been completed.
[0005]
That is, the present invention provides the following general formula (1)
[0006]
[Chemical Formula 3]

[0007]
[Where B isOf cytosine, uracil, 4-N-methylcytosine or 4-N, N-dimethylcytosineRepresents a nucleobase, and Z represents a general formula —CH₂CONR^FourR^Five(R^FourAnd R^FiveAre the same or different, hydrogen atoms, Linear or branched having 1 to 6 carbon atomsAlkylGroup or baseA group represented by R) and R¹, R²And R^ThreeAre the same or different, a hydrogen atom,Represented by general formula (2) belowIndicates a substituted silyl group or an ester-forming residue that can be easily eliminated in vivo.]3′-carbamoylalkyl nucleoside derivative represented by the formula: or a pharmaceutically acceptable salt thereof.
[0008]
The compound of the present invention represented by the above general formula (1) has an excellent antitumor activity and is effective as a pharmaceutical agent such as a therapeutic agent for various tumors.
Accordingly, the present invention also provides a pharmaceutical composition comprising the compound of the above general formula (1) or a pharmaceutically acceptable salt thereof and a pharmaceutical carrier.
Furthermore, the present invention provides a pharmaceutical, particularly an antitumor agent, containing the compound of the above general formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
[0009]
DETAILED DESCRIPTION OF THE INVENTION
  In the above general formula (1), the nucleic acid residue represented by BGroup, CytosineTheRasil4-N-methylcytosine or 4-N, N-dimethylcytosine.
[0015]
  R¹, R²And R^ThreeSubstituted silyl represented byGroupThe following general formula (2)
[0016]
[Formula 4]

[0017]
(Wherein R^a, R^bAnd R^cAre the same or different and each represents a lower alkyl group or a phenyl group)BecauseSpecifically, for example, trimethylsilyl, triethylsilyl, tripropylsilyl, triisopropylsilyl, tritert-butylsilyl, trihexylsilyl, dimethylethylsilyl, dimethylisopropylsilyl, diethylisopropylsilyl, diisopropylmethylsilyl, ditert-butylmethyl A silyl group tri-substituted with a linear or branched alkyl group having 1 to 6 carbon atoms such as silyl, tert-butyldimethylsilyl group, diphenylmethylsilyl, dimethylphenylsilyl, tert-butyldiphenylsilyl, triphenyl A silyl group etc. are mentioned.
[0018]
R¹, R²And R^ThreeAn ester-forming residue which can be easily eliminated in vivo, represented by the following, is easily cleaved in the blood and tissues of mammals including humans and corresponding hydroxyl group compounds (that is, R¹, R²And / or R^ThreeIs a non-toxic ester residue that releases a hydrogen atom, and the ester-forming residue may be any one that protects the hydroxyl group of a well-known nucleoside to form an ester. An acyl group such as an aliphatic acyl group which may have a substituent or an aromatic acyl group which may have a substituent, a lower alkylcarbamoyl group, an amino acid residue and the like.
[0019]
Examples of the aliphatic or aromatic acyl group which may have a substituent include a lower alkanoyl group, an arylcarbonyl group, a heterocyclic carbonyl group, an aryloxycarbonyl group, a lower alkoxycarbonyl group, and an acyloxyacyl group. It is done.
Examples of the lower alkanoyl group include a halogen atom, lower alkoxy as a substituent such as formyl, acetyl, propionyl, butyryl, isobutyryl, pentanoyl, hexanoyl, chloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl, methoxyacetyl, ethoxyacetyl group And an alkanoyl group having 1 to 6 carbon atoms which may have a group or the like.
[0020]
Examples of the arylcarbonyl group include benzoyl, α-naphthylcarbonyl, β-naphthylcarbonyl, 2-methylbenzoyl, 3-methylbenzoyl, 4-methylbenzoyl, 2,4-dimethylbenzoyl, 4-ethylbenzoyl and 2-methoxybenzoyl. 3-methoxybenzoyl, 4-methoxybenzoyl, 2,4-dimethoxybenzoyl, 4-ethoxybenzoyl, 2-methoxy-4-ethoxybenzoyl, 4-propoxybenzoyl, 2-chlorobenzoyl, 3-chlorobenzoyl, 4-chloro Benzoyl, 2,3-dichlorobenzoyl, 2-bromobenzoyl, 4-fluorobenzoyl, 2-carboxybenzoyl, 3-carboxybenzoyl, 4-carboxybenzoyl, 2-cyanobenzoyl, 4-cyanobenzoyl, 2- Benzoyl or naphthylcarbonyl group which may have a lower alkyl group, a lower alkoxy group, a halogen atom, a carboxyl group, a nitro group, a cyano group or the like as a substituent such as a nitrobenzoyl, 4-nitrobenzoyl or 2,4-dinitrobenzoyl group Is mentioned.
[0021]
Examples of the heterocyclic carbonyl group include 2-furanylcarbonyl, 4-thiazolylcarbonyl, 2-quinolylcarbonyl, 2-pyrazinylcarbonyl, 2-pyridylcarbonyl, 3-pyridylcarbonyl, 4-pyridylcarbonyl group and the like. Is mentioned.
Examples of the aryloxycarbonyl group include phenoxycarbonyl, α-naphthyloxycarbonyl, β-naphthyloxycarbonyl, 2-methylphenoxycarbonyl, 3-methylphenoxycarbonyl, 4-methylphenoxycarbonyl, 2,4-dimethylphenoxycarbonyl, 4 -Ethylphenoxycarbonyl, 2-methoxyphenoxycarbonyl, 3-methoxyphenoxycarbonyl, 4-methoxyphenoxycarbonyl, 2,4-dimethoxyphenoxycarbonyl, 4-ethoxyphenoxycarbonyl, 2-methoxy-4-ethoxyphenoxycarbonyl, 2-chloro Phenoxycarbonyl, 3-chlorophenoxycarbonyl, 4-chlorophenoxycarbonyl, 2,3-dichlorophenoxycarbonyl, 2-bromophenoxy Examples include carbonyl, 4-fluorophenoxycarbonyl, β-methyl-α-naphthyloxycarbonyl, β-chloro-α-naphthyloxycarbonyl group and the like.
[0022]
Examples of the lower alkoxycarbonyl group include a methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, isopropoxycarbonyl, n-butoxycarbonyl, isobutoxycarbonyl, sec-butoxycarbonyl, tert-butoxycarbonyl, pentyloxycarbonyl, hexyloxycarbonyl group And a linear or branched alkoxycarbonyl group having 2 to 7 carbon atoms such as
Examples of the acyloxyacyl group include acetyloxyacetyl, propionyloxyacetyl, α- (acetyloxy) propionyl, β- (propionyloxy) propionyl group and the like.
[0023]
Examples of the lower alkylcarbamoyl group include linear or branched alkyl groups having 1 to 6 carbon atoms such as methylcarbamoyl, ethylcarbamoyl, propylcarbamoyl, butylcarbamoyl, pentylcarbamoyl, hexylcarbamoyl, dimethylcarbamoyl, and diethylcarbamoyl groups. And mono- or di-substituted carbamoyl group.
The amino acid residue is a group formed by removing the hydroxyl group from the carboxyl group of the amino acid, and may be derived from any of natural or synthetic amino acids, such as glycine, alanine, β-alanine, Although valine, isoleucine, etc. are mentioned, any may be sufficient as long as it is an amino acid residue of Unexamined-Japanese-Patent No. 1-104093.
[0024]
Other ester-forming residues include, for example, THEODORA W. GREEN, “PROTECTIVE GROUPS IN ORGANIC SYNTHESIS Second Edition”, JOHN WILEY & SONS, INC. (1991), Chemical Society of Japan <New Experimental Chemistry Course 4> “Synthesis and Reaction of Organic Compounds (V)”, Chapter 11 p2495 Maruzen (1983), JP-A 61-106593, JP-A 62-149696, Any of those commonly used as ordinary ester-forming residues described in JP-A-1-153696 may be used.
[0026]
Z is preferably a carbamoylmethyl, methylcarbamoylmethyl, ethylcarbamoylmethyl, benzylcarbamoylmethyl or dimethylcarbamoylmethyl group, more preferably a carbamoylmethyl group.
R¹, R²And R^ThreeExamples of the ester-forming residue which can be easily eliminated in vivo include an acyl group, and more preferably an acetyl group and a benzoyl group.
R¹, R²And R^ThreeAs a preferable example, a hydrogen atom is used.
[0027]
  Preferred compounds of the invention are those wherein B is cytosine, uracil4-N-methylcytosine or 4-N, N-dimethylcytosine, Z is a carbamoylmethyl, methylcarbamoylmethyl, ethylcarbamoylmethyl, benzylcarbamoylmethyl or dimethylcarbamoylmethyl group, R¹, R²And R^ThreeIs a 3′-carbamoylalkyl nucleoside derivative in which is a hydrogen atom.
[0028]
Still more preferred compounds are those in the general formula (1), wherein B is cytosine, uracil, 4-N-methylcytosine or 4-N, N-dimethylcytosine, Z is a carbamoylmethyl group, R¹, R²And R^ThreeIs a 3'-carbamoylalkyl nucleoside derivative in which is a hydrogen atom.
[0029]
The compound of the present invention includes a salt form, and the salt is not particularly limited as long as it is a pharmaceutically acceptable salt.^ThreeIs a hydrogen atom, inorganic acid salts such as hydrochloride, hydrobromide and sulfate, organic sulfonates such as methanesulfonate and benzenesulfonate, acetate, propionate, Examples include acid addition salts such as organic acid salts such as aliphatic carboxylates such as fluoroacetates. The compounds of the present invention also include hydrates thereof.
[0030]
The compound of the present invention represented by the general formula (1) can be produced, for example, according to the following reaction process formulas 1 to 3.
[0031]
[Chemical formula 5]

[0032]
(Where B, R^FourAnd R^FiveIs the same as above. R¹'Represents a substituted silyl group. )
Examples of the substituted silyl group include a group represented by the general formula (2) and a tetraisopropyldisiloxyl (TIPDS) group. Preferred are trimethylsilyl, tert-butyldimethylsilyl, methyldiisopropylsilyl and triisopropylsilyl groups.
[0033]
(Process A)
The lactone compound represented by the general formula (4) is obtained by reacting the compound represented by the general formula (3) with samarium iodide in tetrahydrofuran.
Although the compound represented by General formula (3) is a well-known compound or is obtained by a well-known method, specifically, it can manufacture according to postscript reaction process formula 4. The reaction ratio is 1 to 10 times mol, preferably 1 to 5 times mol of samarium iodide with respect to the compound of general formula (3). The reaction temperature is initially mixed at −78 ° C. and then carried out at 0 to 50 ° C., preferably around room temperature, and the reaction proceeds advantageously in a reaction time of 0.1 to 50 hours, preferably 1 to 24 hours.
[0034]
(Process B)
The compound represented by the general formula (4) obtained in Step A is mixed with R in a suitable solvent.^FourR^FiveBy reacting with an amine compound represented by NH, the compound of the present invention represented by the general formula (1-a) is obtained.
The solvent is not particularly limited as long as it does not participate in the reaction, and any solvent may be used, and examples thereof include lower alcohols such as methanol and ethanol, dimethylformamide and the like.
In this reaction, amines such as dimethylaminopyridine and triethylamine may be added as a catalyst.
[0035]
The reaction rate is preferably 1 to 10 times the molar amount of the amine compound relative to the compound represented by the general formula (4), and the catalyst amount is preferably used when the catalyst is used. The reaction temperature is around -70 ° C when methanolic ammonia is used, and around room temperature when a substituted amine compound is used. The reaction proceeds advantageously in a reaction time of 0.1 to 50 hours, preferably 1 to 24 hours.
[0036]
(Process C)
This invention compound represented by general formula (1-b) is obtained by making the compound represented by general formula (1-a) react with ammonium fluoride in a lower alcohol.
The reaction rate is 5 to 50 times mol, preferably 10 to 30 times mol of ammonium fluoride with respect to the compound represented by the general formula (1-a). The reaction is carried out by heating under reflux, and the reaction proceeds advantageously in a reaction time of 0.1 to 20 hours, preferably 1 to 10 hours.
[0037]
[Chemical 6]

[0038]
(Wherein B and R¹'Is the same as above. R²'And R^Three'Represents a substituted silyl group. ) R²'And R^ThreeAs the silyl group represented by ′, the above R¹The same thing as' is mentioned.
[0039]
(D process)
In the compound represented by the general formula (1-a) obtained in Step B of Reaction Process Formula 1, R^FourAnd R^FiveThe compound of the present invention represented by the general formula (1-c) is obtained by reacting a compound (1-a ′) in which is a hydrogen atom with tert-butyldimethylsilyl chloride in dimethylformamide in the presence of imidazole. obtain.
The proportion of the reaction is 1 to 10 times mol amount, preferably 1 to 5 times mol amount of tert-butyldimethylsilyl chloride, and 1 to 10 times mol of imidazole, relative to the compound represented by the general formula (1-a ′). The amount, preferably 2 to 7 times the molar amount is used. The reaction proceeds advantageously at a reaction temperature of 0 to 100 ° C., preferably around room temperature, and a reaction time of 0.1 to 50 hours, preferably 0.5 to 24 hours.
[0040]
(E process)
The compound represented by the general formula (1-c) obtained in the step D is reacted with p-toluenesulfonyl chloride in pyridine to obtain the present compound represented by the general formula (1-d). .
The proportion of the reaction is 1 to 10 times mol, preferably 1 to 5 times mol, of p-toluenesulfonyl chloride with respect to the compound represented by the general formula (1-c). The reaction is carried out by heating under reflux, and the reaction proceeds advantageously in a reaction time of 0.1 to 50 hours, preferably 0.5 to 24 hours.
[0041]
(F process)
The compound of the present invention represented by the general formula (1-e) is obtained by reacting in the same manner as in Step C of Reaction Step 1.
[0042]
[Chemical 7]

[0043]
(Where Z, R¹'And R^Three'Is the same as above. R⁶And R⁷Are the same or different and each represents a hydrogen atom, a lower alkyl group or a benzyl group which may have a substituent. )
[0044]
(G process)
Among the compounds represented by the general formulas (1-c) and (1-d) obtained by the reaction process formula 2, the compound (1-f) in which B is uracil is converted into 2,4,6- In the presence of triisopropylbenzenesulfonyl, dimethylaminopyridine and triethylamine, R⁶R⁷By reacting with an aqueous amine solution represented by NH, it can be converted to the present compound which is a cytosine derivative represented by the general formula (1-g).
[0045]
The proportion of the reaction is 1 to 10 times, preferably 1 to 5 times, 2,4,6-triisopropylbenzenesulfonyl, dimethyl amine compound with respect to the compound represented by formula (1-f). Aminopyridine and triethylamine are each used in an amount of 1 to 10 times by mole, preferably 1 to 5 times by mole. The reaction temperature is 0 to 100 ° C., preferably around room temperature, and the reaction proceeds advantageously in a reaction time of 0.1 to 100 hours, preferably 1 to 50 hours.
[0046]
The introduction of ester-forming residues into the 2′-position, 3′-position and 5′-position hydroxyl group of the compounds of the general formulas (1-b) and (1-e) obtained in the reaction process formulas 1 to 3 is conventionally known. According to, for example, “PROTECTIVE GROUPS IN ORGANIC SYNTHESIS Second Edition” mentioned above, the Chemical Society of Japan <New Experimental Chemistry Lecture 4> “Synthesis and Reaction of Organic Compounds (V)”, or Japanese Patent Application Laid-Open No. 58-152898. , JP-A-60-56996, JP-A-61-106593, JP-A-62-149696, JP-A-1-153696, etc., leading to other compounds of the present invention. be able to.
[0047]
Further, the compound of the present invention obtained by the above reaction can be converted into a salt form by a conventionally known method such as reacting the compound with the inorganic acid or organic acid in a suitable solvent. Examples of the solvent include water, methanol, ethanol, dichloromethane, tetrahydrofuran, ethyl acetate, hexane and the like. The reaction temperature is preferably 0 to 50 ° C.
Moreover, the raw material compound (3) used by the said reaction process formula 1 is manufactured according to the following reaction process formula 4, for example.
[0048]
[Chemical 8]

[0049]
(Where B, R¹'And R^Three'Is the same as above. )
[0050]
(H process)
The compound represented by the general formula (5) is partially hydrolyzed, for example, J.P. Org. Chem. , 55, 410-412 (1990), that is, by reacting at 0 ° C. in a trifluoroacetic acid-water mixture, selective desilylation reaction at the 5′-position is carried out to give the general formula (6) To obtain a compound represented by:
The compound represented by the general formula (5) is a known compound or a known method, for example, the above-mentioned J.P. Org. Chem. , SYNTHESIS, 283-288 (1991), Tetrahedron 47, 1727-1736 (1991).
[0051]
(Process I)
The compound represented by the general formula (6) obtained in the H step is reacted with bromoacetyl bromide in an appropriate solvent to obtain the compound represented by the general formula (3). The solvent is not particularly limited as long as it does not participate in the reaction, and any solvent may be used, and examples thereof include halogenated hydrocarbons such as chloroform and dichloromethane, dimethylformamide and the like.
In this reaction, amines such as dimethylaminopyridine, triethylamine, 2,6-lutidine and the like may be added as a catalyst.
[0052]
The proportion of the reaction is 1 to 10 times mol amount, preferably 1 to 5 times mol amount of bromoacetyl bromide with respect to the compound represented by the general formula (6), and the catalyst amount is used when using the catalyst. preferable. The reaction temperature is preferably around -78 ° C. The reaction proceeds advantageously in a reaction time of 1 minute to 24 hours, preferably 1 minute to 1 hour.
[0053]
The compound of the present invention and each compound obtained by the above method can be isolated and purified by using generally known separation and purification means such as concentration, solvent extraction, filtration, recrystallization, various chromatography and the like.
[0054]
The compound of the present invention can be made into a pharmaceutical composition according to a usual method using an appropriate pharmaceutical carrier. As the carrier used here, various substances commonly used for ordinary drugs, such as excipients, binders, disintegrants, lubricants, colorants, flavoring agents, flavoring agents, surfactants and the like are used. be able to.
[0055]
The dosage unit form when using the pharmaceutical or pharmaceutical composition of the present invention as a therapeutic agent for tumors of mammals including humans is not particularly limited, and can be appropriately selected according to the therapeutic purpose. Oral preparations such as external preparations (ointments, patches, etc.), parenteral preparations such as aerosols, tablets, coated tablets, powders, granules, capsules, pills, liquids (suspensions, emulsions, etc.) Can be mentioned.
The above various compositions are formulated by a formulation method generally known in this field.
[0056]
In the form of an injection, the carrier may be, for example, a diluent such as water, ethyl alcohol, macrogol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters, citric acid PH adjusting agents and buffers such as sodium, sodium acetate and sodium phosphate, stabilizers such as sodium pyrosulfite, ethylenediaminetetraacetic acid, thioglycolic acid and thiolactic acid can be used. In this case, a sufficient amount of sodium chloride, glucose or glycerin may be contained in the pharmaceutical preparation to prepare an isotonic solution, and a normal solubilizing agent, soothing agent, local anesthetic, etc. may be added. It may be added. By adding these carriers, subcutaneous, intramuscular and intravenous injections can be produced by conventional methods.
[0057]
In the form of a suppository, it is suitable as a carrier, for example, polyethylene glycol, cacao butter, lanolin, higher alcohol, higher alcohol esters, gelatin, semi-synthetic glyceride, Witepsol (registered trademark: Dynamite Nobel), etc. It can be used with the addition of various absorption promoters.
[0058]
When preparing ointments such as pastes, creams and gels, commonly used bases, stabilizers, wetting agents, preservatives, etc. are blended as necessary and mixed and formulated by conventional methods. The As the base, for example, white petrolatum, paraffin, glycerin, cellulose derivative, polyethylene glycol, silicon, bentonite and the like can be used. As preservatives, methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, and the like can be used.
[0059]
When producing a patch, the ointment, cream, gel, paste or the like may be applied to a normal support by a conventional method. As the support, a woven fabric, nonwoven fabric, soft vinyl chloride, polyethylene, polyurethane, or a film or foam sheet made of cotton, suf, or chemical fiber is suitable.
[0060]
In the form of oral solid preparations such as tablets, powders and granules, for example, lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid, methylcellulose, glycerin as carriers Excipients such as sodium alginate and gum arabic, simple syrup, glucose solution, starch solution, gelatin solution, polyvinyl alcohol, polyvinyl ether, polyvinyl pyrrolidone, carboxymethyl cellulose, shellac, methyl cellulose, ethyl cellulose, water, ethanol, potassium phosphate, etc. Binder, dry starch, sodium alginate, agar powder, laminaran powder, sodium bicarbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearin Disintegrating agents such as monoglycerides, starch, lactose, disintegration inhibitors such as sucrose, stearic acid, cocoa butter, hydrogenated oil, quaternary ammonium bases, absorption promoters such as sodium lauryl sulfate, humectants such as glycerin and starch, Adsorbents such as starch, lactose, kaolin, bentonite and colloidal silicic acid, lubricants such as purified talc, stearate, boric acid powder and polyethylene glycol can be used. Further, the tablets can be made into tablets with ordinary coatings as necessary, for example, sugar-coated tablets, gelatin-encapsulated tablets, enteric-coated tablets, film-coated tablets, double tablets, multilayer tablets and the like.
[0061]
Capsules are prepared by mixing with various carriers exemplified above and filling hard gelatin capsules, soft capsules, and the like.
In the form of pills, as carriers, for example, excipients such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, kaolin, talc, binders such as gum arabic powder, tragacanth powder, gelatin, ethanol, laminaran Disintegrants such as agar can be used.
Liquid preparations may be aqueous or oily suspensions, solutions, syrups and elixirs, which are prepared according to conventional methods using conventional additives.
[0062]
The amount of the compound of the present invention to be contained in the preparation varies depending on the dosage form, administration route, administration schedule, etc., and cannot be generally specified, and is appropriately selected from a wide range, but is usually 1 to 70% by weight in the preparation. It is preferable to set the degree.
[0063]
The administration method of the above preparation is not particularly limited, and for example, enteral administration, oral administration, rectal administration, buccal administration, depending on the form of the preparation, the age of the subject to be administered such as a patient, sex and other conditions, the degree of symptoms, etc. Transdermal administration and the like are determined as appropriate. For example, in the case of tablets, pills, solutions, suspensions, emulsions, granules and capsules, it is administered orally, and in the case of suppositories, it is administered rectally. In the case of an injection, it is administered intravenously alone or mixed with a normal fluid such as glucose or amino acid, and further administered alone, if necessary, intraarterially, intramuscularly, intradermally, subcutaneously or intraperitoneally. The ointment is applied to the skin, oral mucosa and the like.
[0064]
The dose of the compound of the present invention is appropriately selected according to the usage, age of the subject to be administered such as a patient, sex, condition, type of tumor, type of the compound of the present invention to be administered, other conditions, etc. It is desirable that the dosage is about 1-1000 mg for oral preparations, about 0.1-500 mg for injections, and about 5-1000 mg for suppositories. The daily dosage of the drug having the above dosage form is usually about 0.1 to 200 mg / kg body weight / day, preferably about 0.5 to 100 mg / kg body weight / day. It is good to do. These preparations of the present invention can be administered once a day or divided into about 2 to 4 times a day.
[0065]
The malignant tumor that can be treated by administering the preparation containing the compound of the present invention is not particularly limited. For example, head and neck cancer, esophageal cancer, stomach cancer, colon cancer, rectal cancer, liver cancer, gallbladder / bile duct cancer, pancreatic cancer Lung cancer, breast cancer, ovarian cancer, bladder cancer, prostate cancer, testicular tumor, bone / soft tissue sarcoma, malignant lymphoma, leukemia, cervical cancer, skin cancer, brain tumor and the like.
[0066]
【The invention's effect】
The 3′-carbamoylalkyl nucleoside derivative of the present invention has an excellent antitumor action and is useful as a medicament for various therapeutic agents for tumors.
[0067]
【Example】
Reference Examples, Examples and Pharmacological Test Examples are shown below to explain the present invention in more detail, but the present invention is not limited by these.
[0068]
Reference example 1
Synthesis of 1- (5-O-bromoacetyl-2-O-tert-butyldimethylsilyl-β-D-erythro-pentofuran-3-urosyl) uracil:
1- (2-O-tert-butyldimethylsilyl-β-D-erythro-pentofuran-3-urosyl) uracil 4.48 g (12.6 mmol) was dissolved in 30 ml of dichloromethane and cooled to -78 ° C under argon atmosphere. 2,6-lutidine (2.20 ml, 17.6 mmol) and bromoacetyl bromide (1.41 ml, 15.1 mmol) were added dropwise and stirred for 5 minutes. To the reaction mixture was added 10 ml of saturated aqueous sodium hydrogen carbonate solution, and the dichloromethane layer was extracted, washed with water (3 × 30 ml) and saturated brine (30 ml), and dried over anhydrous sodium sulfate. After filtration, the filtrate was distilled off under reduced pressure. The residue was dissolved in a small amount of ethyl acetate, adsorbed on silica gel, the solvent was distilled off, and purified by silica gel column chromatography (4 × 20 cm, 30% ethyl acetate / hexane). Thus, 4.12 g (yield 70%) of the title compound was obtained as a yellow foamy substance.
[0069]
FAB-MS: m / z 478 (MH⁺)
¹H-NMR (CDCl_Three) δ: 9.67 (br s, 1H, NH-3, exchanged with D₂O),
7.49 (d, 1H, H-6, J_6,5= 8.2Hz), 5.97 (d, 1H, H-1 ', J_{1 '},_{2 '}= 7.1Hz),
  5.84 (d, 1H, H-5, J_5,6 = 8.2Hz),
4.51 (d, 1H, H-5'a, J_{5'a, 5'b}= 11.9Hz),
4.47 (d, 1H, H-2 ', J_{2 '},_{1 '} = 7.1Hz),
4.44 (dd, 1H, H-4 ', J_Four',_5'a= 2.0, J_Four',_5'b= 3.5Hz),
    4.41 (dd, 1H, H-5b ', J_{5'b, 4 '} = 3.5, J_5'b,_5'a= 11.9Hz),
0.83 (s, 9H, t-Bu), 0.09, 0.02 (each s, each 3H, Me).
¹³C-NMR (CDCl_Three) δ: 206.2, 166.4, 163.2, 150.6, 140.3, 104.1,
87.93, 78.77, 75.35, 64.54, 25.81, 25.64,
25.54, 18.33, -4.4, -5.01.
[0070]
Reference example 2
Lactone compound represented by the general formula (4) (B is uracil, R¹′ Is the synthesis of tert-butyldimethylsilyl):
After cooling 14.2 ml (1.24 mmol) of 0.1N samarium iodide-tetrahydrofuran solution to −78 ° C. under an argon atmosphere, 338 mg (0.567 mmol) of the compound obtained in Reference Example 1 dissolved in 6 ml of tetrahydrofuran was slowly added dropwise. . The reaction mixture was returned to room temperature, stirred with 4 ml of 1N hydrochloric acid, extracted with 50 ml of ethyl acetate, washed with water (2 × 20 ml) and saturated brine (20 ml), and dried over anhydrous sodium sulfate. After filtration, the filtrate was distilled off under reduced pressure. The residue was dissolved in a small amount of ethyl acetate, adsorbed on silica gel, the solvent was distilled off, and purified by silica gel column chromatography (2 × 11 cm, 30% ethyl acetate / hexane). Thus, 219 mg (yield 75%) of the title lactone was obtained as a yellowish white glassy substance. Crystallization from ethyl acetate-hexane as an analytical sample gave pale yellow crystals.
[0071]
FAB-MS: m / z 399 (MH⁺)
¹H-NMR (CDCl_Three) δ: 8.19 (br s, 1H, NH-3, exchanged with D₂O),
7.16 (d, 1H, H-6, J_6,5= 8.1Hz),
5.98 (d, 1H, H-1 ', J_{1 '},_{2 '} = 7.5Hz),
5.84 (d, 1H, H-5, J_5,6= 8.1Hz),
4.46 (d, 1H, H-5'a, J_5'a,_5'b= 13.1Hz), 4.41 (br s, 1H, H-4 '),
4.37 (dd, 1H, H-5'b, J_5'b,_Four' = 2.4, J_5'b,_5'a= 13.1Hz),
3.66 (d, 1H, H-2 ', J_{2 '},_{1 '} = 7.5Hz),
3.49 (s, 1H, 3'-OH, exchanged with D₂O),
    2.78, 2.71 (each d, each 1H, H-3``a, H-3''b, J = 15.3Hz),
0.91 (s, 9H, t-Bu), 0.11, -0.02 (each s, each 3H, Me).
<sup>13</sup>C-NMR (CDCl<sub>Three</sub>) δ: 169.6, 162.4, 150.3, 131.6, 104.4, 85.02,
81.39, 74.08, 68.36, 40.63, 25.68, 17.97, 11.68, -4.52, -4.64.
[0072]
Example 1
Synthesis of 1- (2-O-tert-butyldimethylsilyl-3-C-carbamoylmethyl-β-D-ribofuranosyl) uracil (compound 1):
The lactone compound 808 mg (2.03 mmol) obtained in Reference Example 2 was dissolved in 5 ml of methanol, 20 ml of a saturated ammonia methanol solution cooled to −70 ° C. was added, and the mixture was allowed to stand at −70 ° C. for 3 hours. The reaction mixture was evaporated under reduced pressure, the residue was dissolved in a small amount of methanol, adsorbed onto silica gel, the solvent was distilled off, and the residue was purified by silica gel column chromatography (4 × 10 cm). 98%) was obtained as a white powder. Crystallization from methanol / chloroform as a sample for analysis gave pale yellow needles.
[0073]
FAB-MS: m / z 415 (MH<sup>+</sup>)
Elemental analysis:
Calculated value (C<sub>17</sub>H<sub>29</sub>N<sub>Three</sub>O<sub>7</sub>As Si): C, 49.26; H, 6.81; N, 10.14.
Found: C, 49.10; H, 7.02; N, 10.13.
[0074]
Example 2
Synthesis of 1- (2,5-O-di-tert-butyldimethylsilyl-3-C-carbamoylmethyl-β-D-ribofuranosyl) uracil (compound 2):
1.50 g (3.62 mmol) of Compound 1 obtained in Example 1 was dissolved in 50 ml of dimethylformamide, 738 mg (10.7 mmol) of imidazole and 819 mg (5.43 mmol) of tert-butyldimethylsilyl chloride were added, and the mixture was stirred at room temperature for 12 hours. . Ethanol (5 ml) was added to the reaction mixture and stirred for 15 minutes, and the solvent was evaporated under reduced pressure. The residue was partitioned by adding 100 ml of ethyl acetate and 100 ml of water, and the ethyl acetate layer was washed with water (3 × 50 ml) and saturated brine (50 ml) and then dried over anhydrous sodium sulfate. After filtration, the filtrate was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (4 × 8.5 cm, 4% methanol / chloroform) to give 1.9 g (99% yield) of the title compound 2 as a yellow foam. Obtained.
[0075]
FAB-MS: m / z 529 (MH<sup>+</sup>)
<sup>1</sup>H-NMR (CDCl<sub>Three</sub>) δ: 8.14 (br s, 1H, NH-3, exchanged with D<sub>2</sub>O),
8.00 (d, 1H, H-6, J<sub>6,5</sub>= 8.2Hz),
6.88 (br s, 1H, amide, exchanged with D<sub>2</sub>O),
6.17 (d, 1H, H-1 ', J<sub>1 '</sub>,<sub>2 '</sub> = 7.2Hz),
5.74 (dd, 1H, H-5, J<sub>5,6</sub>= 8.2Hz, J<sub>Five</sub>,<sub>NH-3</sub> = 2.0Hz),
5.42 (br s, 1H, amide, exchanged with D<sub>2</sub>O),
4.27 (br s, 1H, H-4 '), 4.07 (d, 1H, H-2', J<sub>2 '</sub>,<sub>1 '</sub> = 7.2Hz),
3.91 (dd, 1H, H-5'a, J<sub>5'a</sub>,<sub>Four'</sub> = 2.1, J<sub>5'a</sub>,<sub>5'b</sub>= 12.1Hz),
3.87 (dd, 1H, H-5'b, J<sub>5'b</sub>,<sub>Four'</sub> = 2.2, J<sub>5'b</sub>,<sub>5'a</sub>= 12.1Hz),
3.47 (s, 1H, 3'-OH, exchanged with D<sub>2</sub>O),
2.64, 2.56 (each d, each 1H, H-3``a, H-3''b, J = 15.9Hz),
0.97, 0.91 (each s, each 9H, t-Bu),
0.23, 0.22, 0.06, -0.05 (each s, each 3H, Me).
¹³C-NMR (CDCl_Three) δ: 172.3, 163.19, 151.0, 140.54, 103.49, 86.12,
85.10, 78.84, 78.31, 63.31, 40.68, 26.24, 25.80, 18.54, 18.00,
11.69, -4.35, -4.50, -5.10, -5.52.
[0076]
Example 3
Synthesis of 1- (2,5-O-di-tert-butyldimethylsilyl-3-C-carbamoylmethyl-β-D-ribofuranosyl) cytosine (compound 3):
1.05 g (2 mmol) of Compound 2 obtained in Example 2 was dissolved in 50 ml of acetonitrile under an argon atmosphere, 1.21 g (4.00 mmol) of 2,4,6-triisopropylbenzenesulfonyl chloride, 487 mg (4.00 mmol) of dimethylaminopyridine. And 0.28 ml (4.00 mmol) of triethylamine were added and stirred at room temperature for 24 hours, concentrated aqueous ammonia (28%, 20 ml) was added, and the mixture was further stirred at room temperature for 2 hours. The reaction mixture was evaporated under reduced pressure, the residue was dissolved in a small amount of methanol, adsorbed on silica gel, evaporated to remove the solvent, and purified by silica gel column chromatography (2 × 18 cm, 20% methanol / chloroform). The title compound 3 was obtained as white crystals (830 mg, yield 76%). Recrystallization from methanol / chloroform as a sample for analysis gave white plate crystals.
[0077]
  FAB-MS: m / z 530 (MH⁺)
¹H-NMR (CDCl_Three) δ: 8.04 (d, 1H, H-6, J_6,5= 8.2Hz),
7.23 (br s, 1H, amide a, exchanged with D₂O),
6.31 (d, 1H, H-1 ', J_{1 '},_{2 '} = 7.2Hz), 5.95 (br s, 1H, H-5),
5.49 (br s, 1H, amide b, exchanged with D₂O),
4.24 (br s, 1H, H-4 '), 4.06 (d, 1H, H-2', J_{2 ', 1'}= 7.2Hz),
4.01 (d, 1H, H-5'a, J_{5'a, 5'b}= 11.8Hz),
3.89 (dd, 1H, H-5'b, J_5'b,_Four' = 2.0, J_{5'b, 5'a}= 12.2Hz),
2.59 (t, 2H, H-3``a, H-3''b, J = 15.1Hz),
0.96, 0.85 (each s, each 9H, t-Bu),
0.17, -0.01, -0.10 (each s, each 3H, Me).
[0082]
Example 6
Synthesis of 1- (3-C-carbamoylmethyl-β-D-ribofuranosyl) uracil (Compound 6):
281 mg (0.678 mmol) of Compound 1 obtained in Example 1 was dissolved in 20 ml of methanol, 502 mg (13.6 mmol) of ammonium fluoride was added, and the mixture was heated to reflux for 2 hours. The reaction mixture was evaporated under reduced pressure, and the residue was partitioned by adding 50 ml of water and 50 ml of chloroform. The aqueous layer was further washed with chloroform (2 × 30 ml) and concentrated to 20 ml under reduced pressure. Activated charcoal was added until the water disappeared, and the column was packed into a column, washed with 100 ml of water, and eluted with 50% aqueous methanol. The eluate was concentrated under reduced pressure to obtain 205 mg (yield 99%) of the title compound 6 as a white powder. Crystallization from water / methanol as an analytical sample gave white needle crystals.
[0083]
FAB-MS: m / z 302 (MH<sup>+</sup>)
<sup>1</sup>H-NMR (DMSO-d<sub>6</sub>) δ: 9.45 (br s, 1H, NH-3, exchanged with D<sub>2</sub>O),
8.05 (d, 1H, H-6, J<sub>6,5</sub> = 8.0Hz),
7.47, 7.06 (each br s, each 1H, amide, exchanged with D<sub>2</sub>O),
5.91 (d, 1H, H-1 ', J<sub>1 '</sub>,<sub>2 '</sub> = 7.9Hz),
    5.67 (d, 1H, H-5, J<sub>5,6</sub>= 8.1Hz),
5.56 (d, 1H, 2'-OH, J<sub>2'-OH, 2 '</sub> = 6.1Hz, exchanged with D<sub>2</sub>O),
5.23 (t, 1H, 5'-OH, J<sub>5'-OH, 5 '</sub> = 3.9Hz, exchanged with D<sub>2</sub>O),
5.19 (s, 1H, 3'-OH, exchanged with D<sub>2</sub>O), 3.96 (br s, 1H, H-4 '),
3.94 (dd, 1H, H-2 ', J<sub>2 ', 2'-OH</sub> = 6.3, J<sub>2 ', 1'</sub>= 7.8Hz),
3.71 (ddd, 1H, H-5'a, J<sub>5'a, 4 '</sub> = 1.5,
J<sub>5'a, 5'b</sub>= 12.0, J<sub>5'a, 5'-OH</sub>= 3.4Hz),
3.59 (ddd, 1H, H-5'b, J<sub>5'b, 4 '</sub> = 3.0,
J<sub>5'b, 5'a</sub>= 12.0, J<sub>5'b, 5'-OH</sub>= 3.4Hz),
2.57, 2.50 (each d, each 1H, H-3``a, 3''b, J = 15.3Hz).
[0084]
Example 7
Synthesis of 1- (3-C-carbamoylmethyl-β-D-ribofuranosyl) cytosine (Compound 7):
105 mg (0.2 mmol) of the compound 3 obtained in Example 3 was dissolved in 5 ml of methanol, 148 mg (4.00 mmol) of ammonium fluoride was added, and the mixture was heated to reflux for 2 hours. The reaction mixture was evaporated under reduced pressure, the residue was partitioned by adding 10 ml of water and 10 ml of chloroform, and the aqueous layer was washed with chloroform (2 × 10 ml) and concentrated to 5 ml under reduced pressure. Activated charcoal was added until there was no more, and it was packed in a column, washed with 50 ml of water, and eluted with 50% aqueous methanol. The eluate was concentrated under reduced pressure to obtain 47.5 mg (yield 79%) of the title compound 7 as a white powder. A white powder was obtained by crystallization from water / methanol as an analytical sample.
[0085]
FAB-MS: m / z 301 (MH⁺)
¹H-NMR (DMSO-d₆) δ: 7.90 (d, 1H, H-6, J_6,5= 7.4Hz),
7.47 (br s, 1H, amide a, exchanged with D₂O),
7.17 (br s, 2H, NH₂, exchanged with D₂O),
7.05 (br s, 1H, amide b, exchanged with D₂O),
5.88 (d, 1H, H-1 ', J_{1 ', 2'}= 7.9Hz), 5.75 (br s, 1H, H-5)_,
    5.41 (d, 1H, 2'-OH, J_{2'-OH, 2 '} = 6.2Hz, exchanged with D₂O),
5.22 (t, 1H, 5'-OH, J_{5'-OH, 5 '} = 4.5Hz, exchanged with D₂O),
5.13 (s, 1H, 3'-OH, exchanged with D₂O),
3.98 (dd, 1H, H-2 ', J_{2 ', 2'-OH} = 7.1, J_{2 ', 1'}= 7.1Hz),
3.93 (br s, 1H, H-4 '),
3.66 (ddd, 1H, H-5'a,
J_{5'a, 4 '} = 2.9, J_{5'a, 5'b}= 9.8, J_{5'a, 5'-OH}= 4.5Hz),
3.57 (ddd, 1H, H-5'b, J_{5'b, 4 '} = 2.4,
J_{5'b, 5'a}= 9.7, J_{5'b, 5'-OH}= 4.5Hz),
2.55, 2.49 (each d, each 1H, H-3``a, 3''b, J = 16.3Hz).
[0090]
Example 10
Synthesis of 1- [2-O-tert-butyldimethylsilyl-3-C- (N-methylcarbamoylmethyl) -β-D-ribofuranosyl] uracil (Compound 10):
The lactone compound obtained in Reference Example 2 (398 mg, 1 mmol) was dissolved in dimethylformamide (5 ml), methylamine hydrochloride (333 mg, 5 mmol) and triethylamine (700 μl, 5.00 mmol) were added, and the mixture was stirred at room temperature for 7 hours. The reaction mixture was evaporated under reduced pressure, the residue was partitioned between 100 ml of ethyl acetate and 50 ml of water, and the ethyl acetate layer was further washed with water (3 × 50 ml) and saturated brine (50 ml) and then dried over anhydrous sodium sulfate. . After filtration, the filtrate was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (2 × 6 cm, 4% methanol / chloroform) to obtain 456 mg (yield 99%) of the title compound 10 as a yellow glassy substance. .
[0091]
FAB-MS: m / z 430 (MH<sup>+</sup>)
<sup>1</sup>H-NMR (CDCl<sub>Three</sub>) δ: 8.15 (br s, 1H, NH-3, exchanged with D<sub>2</sub>O),
7.70 (d, 1H, H-6, J<sub>6,5</sub>= 8.2Hz),
6.60 (d, 1H, amide, J<sub>NH, Me</sub>= 4.7Hz, exchanged with D<sub>2</sub>O),
5.79 (dd, 1H, H-5, J<sub>5,6</sub>= 8.2, J<sub>5, NH-3</sub> = 2.2Hz),
6.17 (d, 1H, H-1 ', J<sub>1 ', 2'</sub>= 7.3Hz),
4.07 (d, 1H, H-2 ', J<sub>2 ', 1'</sub>= 7.3Hz),
4.27 (br s, 1H, H-4 '),
3.88 (dd, 1H, H-5'a, J<sub>5'a, 4 '</sub> = 2.4, J<sub>5'a, 5'b</sub>= 12.6Hz),
3.76 (dd, 1H, H-5'b, J<sub>5'b, 4 '</sub> = 1.2, J<sub>5'b, 5'a</sub>= 12.6Hz),
3.54 (s, 1H, 3'-OH, exchanged with D<sub>2</sub>O),
2.83 (d, 3H, Me, J<sub>Me, NH</sub>= 4.7Hz),
2.80, 2.56 (each d, each 1H, H-3``a, H-3''b, J = 15.7Hz),
0.89 (s, 9H, t-Bu), 0.63, -0.01 (each s, each 3H, Me).
[0092]
Example 11
Synthesis of 1- [2-O-tert-butyldimethylsilyl-3-C- (N, N-dimethylcarbamoylmethyl) -β-D-ribofuranosyl] uracil (Compound 11):
In the same manner as in Example 10 from 398 mg (1 mmol) of the lactone compound obtained in Reference Example 2, the title compound 11 was obtained in a yield of 99% as a yellow glassy substance.
[0093]
FAB-MS: m / z 444 (MH⁺)
¹H-NMR (CDCl_Three) δ: 8.19 (br s, 1H, NH-3, exchanged with D₂O),
7.32 (d 1H, H-6, J_6,5= 8.0Hz),
5.77 (dd, 1H, H-5, J_5,6= 8.0, J_{5, NH-3} = 2.2Hz),
5.41 (d, 1H, H-1 ', J_{1 ', 2'}= 7.7Hz),
    4.75 (d, 1H, H-2 ', J_{2 ', 1'}= 7.7Hz),
4.39 (br s, 1H, H-4 '),
3.83 (dd, 1H, H-5'a, J_{5'a, 4 '} = 2.3, J_{5'a, 5'b}= 13.1Hz),
3.57 (dd, 1H, H-5'b, J_{5'b, 4 '} = 1.0, J_{5'b, 5'a}= 1.3Hz),
3.03, 2.97 (each s, each 3H, Me),
2.99, 2.75 (each d, each 1H, H-3``a, H-3''b, J = 16.5Hz),
0.89 (s, 9H, t-Bu), 0.86, -0.06 (each s, each 3H, Me).
[0094]
Example 12
Synthesis of 1- [2-O-tert-butyldimethylsilyl-3-C- (N-ethylcarbamoylmethyl) -β-D-ribofuranosyl] uracil (Compound 12):
In the same manner as in Example 10 from 398 mg (1 mmol) of the lactone compound obtained in Reference Example 2, the title compound 12 was obtained in a yield of 99% as a yellow glassy substance.
[0095]
FAB-MS: m / z 444 (MH<sup>+</sup>)
<sup>1</sup>H-NMR (CDCl<sub>Three</sub>) δ: 9.28 (br s, 1H, NH-3, exchanged with D<sub>2</sub>O),
7.78 (d, 1H, H-6, J<sub>6,5</sub>= 8.1Hz),
6.80 (d, 1H, amide, J<sub>NH, CH2</sub> = 5.3Hz, exchanged with D<sub>2</sub>O),
5.79 (d, 1H, H-1 ', J<sub>1 ', 2'</sub>= 7.3Hz),
5.71 (dd, 1H, H-5, J<sub>5,6</sub>= 8.1, J<sub>5, NH-3</sub> = 2.2Hz),
4.40 (d, 1H, H-2 ', J<sub>2 ', 1'</sub>= 7.3Hz),
4.20 (br s, 1H, 5'-OH, exchanged with D<sub>2</sub>O),
4.17 (s, 1H, 3'-OH, exchanged with D<sub>2</sub>O),
4.01 (br s, 1H, H-4 '),
3.80 (br d, 1H, H-5'a, J<sub>5'a, 4 '</sub> = 1.0, J<sub>5'a, 5'b</sub>= 12.1Hz),
3.73 (dd, 1H, H-5'b, J<sub>5'b, 4 '</sub> = 5.0, J<sub>5'b, 5'a</sub>= 12.1Hz),
3.26 (dq, 2H, CH<sub>2</sub>CH<sub>Three</sub>, J<sub>CH2, NH</sub> = 5.3, J = 7.1Hz),
2.73, 2.51 (each d, each 1H, H-3``a, H-3''b, J = 15.7Hz),
1.12 (t, 1H, Me, J = 7.1Hz), 0.89 (s, 9H, t-Bu),
    0.63, -0.01 (each s, each 3H, Me).
[0096]
Example 13
  Synthesis of 1- [2-O-tert-butyldimethylsilyl-3-C- (N-benzylcarbamoylmethyl) -β-D-ribofuranosyl] uracil (Compound 13):
In the same manner as in Example 10 from 398 mg (1 mmol) of the lactone compound obtained in Reference Example 2, the title compound 13 was obtained as a colorless glassy substance in a yield of 80%.
[0097]
FAB-MS: m / z 506 (MH⁺)
¹H-NMR (CDCl_Three) δ: 8.10 (br s, 1H, NH-3, exchanged with D₂O),
8.05 (d, 1H, H-6, J_6,5= 8.3Hz), 7.37-7.33 (m, 5H, aromatic),
7.15 (t, 1H, amide, J_{NH, CH}= 5.6Hz, exchanged with D₂O),
6.15 (d, 1H, H-1 ', J_{1 ', 2'}= 7.3Hz),
5.72 (dd, 1H, H-5, J_5,6= 8.1, J_{5, NH-3} = 2.3Hz),
4.56 (d, 2H, PhCH₂, J = 5.6Hz), 4.19 (br s, 1H, H-4 '),
4.07 (d, 1H, H-2 ', J_{2 ', 1'}= 7.3Hz),
3.87 (dd, 1H, H-5'a, J_{5'a, 4 '} = 1.2, J_{5'a, 5'b}= 12.5Hz),
3.82 (dd, 1H, H-5'b, J_{5'b, 4 '} = 2.1, J_{5'b, 5'a}= 12.5Hz),
3.40 (s, 1H, 3'-OH, exchanged with D₂O),
2.68, 2.44 (each d, each 1H, H-3``a, H-3''b, J = 15.5Hz),
0.96, 0.89 (each s, each 9H, t-Bu),
0.18, 0.17, 0.03, -0.07 (each s, each 3H, Me).
[0098]
Example 14
Synthesis of 1- [2,5-O-di-tert-butyldimethylsilyl-3-C- (N-methylcarbamoylmethyl) -β-D-ribofuranosyl] uracil (compound 14):
300 mg (0.7 mmol) of the compound 10 obtained in Example 10 was dissolved in 20 ml of dimethylformamide, 190 mg (2.8 mmol) of imidazole and 211 mg (1.4 mmol) of tert-butyldimethylsilyl chloride were added, and the mixture was stirred at room temperature for 1 hour. 2 ml of ethanol was added to the reaction mixture and stirred for 15 minutes, and the solvent was distilled off under reduced pressure. The residue was partitioned by adding 50 ml of ethyl acetate and 20 ml of water, and the ethyl acetate layer was washed with water (3 × 20 ml) and saturated brine (20 ml) and then dried over anhydrous sodium sulfate. After filtration, the filtrate was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (2 × 6 cm, 40% ethyl acetate / hexane) to obtain 398 mg (yield 100%) of the title compound 14 as a white foam. It was.
[0099]
FAB-MS: m / z 544 (MH<sup>+</sup>)
<sup>1</sup>H-NMR (CDCl<sub>Three</sub>) δ: 8.06 (br s, 1H, NH-3, exchanged with D<sub>2</sub>O),
8.03 (d, 1H, H-6, J<sub>6,5</sub>= 8.2Hz),
6.79 (d, 1H, amide, J<sub>NH, Me</sub>= 4.7Hz, exchanged with D<sub>2</sub>O),
6.16 (d, 1H, H-1 ', J<sub>1 ', 2'</sub>= 7.2Hz),
5.73 (dd, 1H, H-5, J<sub>5,6</sub>= 8.1, J<sub>5, NH-3</sub> = 2.2Hz),
4.22 (br s, 1H, H-4 '), 4.06 (d, 1H, H-2', J<sub>2 ', 1'</sub>= 7.2Hz),
3.88 (br s, 2H, H-5'a, b),
3.52 (s, 1H, 3'-OH, exchanged with D<sub>2</sub>O),
2.81 (d, 3H, Me, J<sub>Me, NH</sub>= 4.7Hz),
2.63, 2.51 (each d, each 1H, H-3``a, H-3''b, J = 15.5Hz),
0.96, 0.89 (each s, each 9H, t-Bu),
0.18, 0.10, 0.03, -0.06 (each s, each 3H, Me).
[0100]
Example 15
Synthesis of 1- [2,5-O-di-tert-butyldimethylsilyl-3-C- (N, N-dimethylcarbamoylmethyl) -β-D-ribofuranosyl] uracil (Compound 15):
In the same manner as in Example 14, from 192 mg (0.44 mmol) of Compound 11 obtained in Example 11, the title compound 15 was obtained as a white foam in a yield of 85%.
[0101]
FAB-MS: m / z 558 (MH⁺)
¹H-NMR (CDCl_Three) δ: 7.97 (br s, 1H, NH-3, exchanged with D₂O),
7 .88 (d, 1H, H-6, J_6,5= 8.1Hz),
    6.23 (d, 1H, H-1 ', J_{1 ', 2'}= 7.5Hz),
5.71 (dd, 1H, H-5, J_5,6= 8.1, J_{5, NH-3} = 2.3Hz),
4.46 (br s, 1H, H-4 '), 4.04 (d, 1H, H-2', J_{2 ', 1'}= 7.4Hz),
3.88 (dd, 1H, H-5'a, J_{5'a, 4 '} = 1.2, J_{5'a, 5'b}= 12.0Hz),
3.86 (dd, 1H, H-5'b, J_{5'b, 4 '} = 2.4, J_{5'b, 5'a}= 12.0Hz),
3.51 (s, 1H, 3'-OH, exchanged with D₂O),
3.00, 2.98 (each s, each 3H, Me),
2.80, 2.71 (each d, each 1H, H-3``a, H-3''b, J = 16.3Hz),
0.95, 0.90 (each s, each 9H, t-Bu),
0.12, 0.11, 0.04, -0.07 (each s, each 3H, Me)<sub>.</sub>
[0102]
Example 16
Synthesis of 1- [2,5-O-di-tert-butyldimethylsilyl-3-C- (N-ethylcarbamoylmethyl) -β-D-ribofuranosyl] uracil (Compound 16):
257 mg (yield 92%) of the title compound 16 was obtained as a white foam from 222 mg (0.5 mmol) of the compound 12 obtained in Example 12 in the same manner as in Example 14.
[0103]
FAB-MS: m / z 558 (MH<sup>+</sup>)
<sup>1</sup>H-NMR (CDCl<sub>Three</sub>) δ: 7.97 (br s, 1H, NH-3, exchanged with D<sub>2</sub>O),
7.88 (d, 1H, H-6, J<sub>6,5</sub>= 8.1Hz),
6.23 (d, 1H, H-1 ', J<sub>1 ', 2'</sub>= 7.5Hz),
5.71 (dd, 1H, H-5, J<sub>5,6</sub>= 8.1, J<sub>5, NH-3</sub> = 2.3Hz),
4.46 (br s, 1H, H-4 '), 4.04 (d, 1H, H-2', J<sub>2 ', 1'</sub>= 7.4Hz),
3.88 (dd, 1H, H-5'a, J<sub>5'a, 4 '</sub> = 1.2, J<sub>5'a, 5'b</sub>= 12.0Hz),
3.86 (dd, 1H, H-5'b, J<sub>5'b, 4 '</sub> = 2.4, J<sub>5'b, 5'a</sub>= 12.0Hz),
3.51 (s, 1H, 3'-OH, exchanged with D<sub>2</sub>O),
3.00, 2.98 (each s, each 3H, Me),
2.80, 2.71 (each d, each 1H, H-3``a, H-3''b, J = 16.3Hz),
0.95, 0.90 (each s, each 9H, t-Bu),
    0.12, 0.11, 0.04, -0.07 (each s, each 3H, Me).
[0104]
Example 17
Synthesis of 1- [2,5-O-di-tert-butyldimethylsilyl-3-C- (N-benzylcarbamoylmethyl) -β-D-ribofuranosyl] uracil (Compound 17):
460 mg (93% yield) of the title compound 17 was obtained as a white foam from 404 mg (0.8 mmol) of the compound 13 obtained in Example 13 in the same manner as in Example 14.
[0105]
FAB-MS: m / z 620 (MH⁺)
¹H-NMR (CDCl_Three) δ: 8.06 (br s, 1H, NH-3, exchanged with D₂O),
8.01 (d, 1H, H-6, J_6,5= 8.3Hz), 7.37-7.26 (m, 5H, aromatic),
7.13 (t, 1H, amide, J_{NH, CH}= 5.7Hz, exchanged with D₂O),
6.15 (d, 1H, H-1 ', J_{1 ', 2'}= 7.3Hz),
5.72 (dd, 1H, H-5, J_5,6= 8.1, J_{5, NH-3} = 2.3Hz),
4.46-4.45 (each d, each 1H, PhCH₂, J = 5.8Hz),
4.19 (br s, 1H, H-4 '), 4.07 (d, 1H, H-2', J_{2 ', 1'}= 7.3Hz),
3.87 (dd, 1H, H-5'a, J_{5'a, 4 '} = 1.2, J_{5'a, 5'b}= 12.5Hz),
3.82 (dd, 1H, H-5'b, J_{5'b, 4 '} = 2.1, J_{5'b, 5'a}= 12.5Hz),
3.40 (s, 1H, 3'-OH, exchanged with D₂O),
2.68, 2.44 (each d, each 1H, H-3``a, H-3''b, J = 15.5Hz),
0.96, 0.89 (each s, each 9H, t-Bu),
0.18, 0.17, 0.03, -0.07 (each s, each 3H, Me).
[0106]
Example 18
Synthesis of 1- [2,5-O-di-tert-butyldimethylsilyl-3-C- (N-methylcarbamoylmethyl) -β-D-ribofuranosyl] cytosine (Compound 18):
217 mg (0.4 mmol) of the compound 14 obtained in Example 14 was dissolved in 10 ml of acetonitrile under an argon atmosphere, 242 mg (0.8 mmol) of 2,4,6-triisopropylbenzenesulfonyl chloride, 97 mg (0.8 mmol) of dimethylaminopyridine. And 0.056 ml (0.8 mmol) of triethylamine were added and stirred at room temperature for 24 hours, and then concentrated aqueous ammonia (28%<sub>,</sub>10 ml) was added and the mixture was further stirred at room temperature for 2 hours. The reaction mixture was evaporated under reduced pressure, the residue was dissolved in a small amount of methanol, adsorbed on silica gel, the solvent was distilled off, and purified by silica gel column chromatography (2 × 8 cm, 10% methanol / chloroform). The title compound 18 was obtained as white crystals (157 mg, yield 73%). Recrystallization from methanol / chloroform as a sample for analysis gave white plate crystals.
[0107]
FAB-MS: m / z 543 (MH<sup>+</sup>)
<sup>1</sup>H-NMR (CDCl<sub>Three</sub>) δ: 8.06 (d, 1H, H-6, J<sub>6,5</sub>= 8.2Hz),
7.05 (d, 1H, amide, J<sub>NH, Me</sub>= 4.5Hz, exchanged with D<sub>2</sub>O),
6.23 (d, 1H, H-1 ', J<sub>1 ', 2'</sub>= 6.7Hz), 6.15 (br s, 1H, H-5),
4.19 (br s, 1H, H-4 '), 4.05 (d, 1H, H-2', J<sub>2 ', 1'</sub>= 6.7Hz),
3.99 (d, 1H, H-5'a, J<sub>5'a, 5'b</sub>= 11.5Hz),
3.87 (d, 1H, H-5'b, J<sub>5'b, 5'a</sub>= 11.5Hz),
2.80 (d, 3H, Me, J<sub>Me, NH</sub>= 4.5Hz),
2.60, 2.53 (each d, each 1H, H-3``a, H-3''b, J = 15.1Hz),
0.95, 0.84 (each s, each 9H, t-Bu),
0.17, 0.16, -0.01, -0.03 (each s, each 3H, Me).
[0108]
Example 19
Synthesis of 1- [2,5-O-di-tert-butyldimethylsilyl-3-C- (N, N-dimethylcarbamoylmethyl) -β-D-ribofuranosyl] cytosine (Compound 19):
115 mg (yield 69%) of the title compound 19 was obtained as a white powder from 167 mg (0.3 mmol) of the compound 15 obtained in Example 15 in the same manner as in Example 18.
[0109]
FAB-MS: m / z 557 (MH⁺)
¹H-NMR (CDCl_Three) δ: 8.15 (d, 1H, H-6, J_6,5= 6.5Hz),
6.30 (br s, 1H, H-5), 6.23 (d, 1H, H-1 ', J_{1 ', 2'}= 7.3Hz),
4.43 (br s, 1H, H-4 '), 4.03 (d, 1H, H-2', J_{2 ', 1'}= 7.3Hz),
    3.87 (br s, 2H, H-5'a, b), 2.98, 2.96 (each s, each 3H, Me),
2.76, 2.74 (each d, each 1H, H-3``a, H-3''b, J = 16.8Hz),
0.93, 0.88 (each s, each 9H, t-Bu),
0.12, 0.10, 0.02, -0.01 (each s, each 3H, Me).
[0110]
Example 20
Synthesis of 1- [2,5-O-di-tert-butyldimethylsilyl-3-C- (N-ethylcarbamoylmethyl) -β-D-ribofuranosyl] cytosine (Compound 20):
In the same manner as in Example 18, 280 mg (yield 100%) of the title compound 20 was obtained as a colorless glassy substance from 280 mg (0.5 mmol) of the compound 16 obtained in Example 16.
[0111]
FAB-MS: m / z 557 (MH<sup>+</sup>)
<sup>1</sup>H-NMR (CDCl<sub>Three</sub>) δ: 8.02 (d, 1H, H-6, J<sub>6,5</sub>= 7.5Hz),
7.10 (br s, 1H, amide, exchanged with D<sub>2</sub>O),
6.31 (d, 1H, H-5, J<sub>5,6</sub>= 7.5Hz),
6.31 (d, 1H, H-1 ', J<sub>1 ', 2'</sub>= 7.1Hz),
4.15 (br s, 1H, H-4 '), 4.05 (d, 1H, H-2', J<sub>2 ', 1'</sub>= 7.1Hz),
3.95 (d, 1H, H-5'a, J<sub>5'a, 5'b</sub>= 11.8Hz),
3.85 (dd, 1H, H-5'b, J<sub>5'b, 4 '</sub> = 11.8Hz),
3.27 (dq, 2H, NCH<sub>2</sub> , J = 7.2Hz),
2.59, 2.52 (each d, each 1H, H-3``a, H-3''b, J = 16.8Hz),
1.13 (t, 3H, NCH₂CH_Three, J = 7.2Hz),
0.95, 0.86 (each s, each 9H, t-Bu),
0.16, 0.15, -0.04, -0.10 (each s, each 3H, Me).
[0112]
Example 21
Synthesis of 1- [2,5-O-di-tert-butyldimethylsilyl-3-C- (N-benzylcarbamoylmethyl) -β-D-ribofuranosyl] cytosine (Compound 21):
352 mg (yield: 89.0%) of the title compound 21 was obtained as a white powder from 402 mg (0.640 mmol) of the compound 17 obtained in Example 17 in the same manner as in Example 18.
[0113]
FAB-MS: m / z 619 (MH⁺)
¹H-NMR (CDCl_Three) δ: 8.03 (d, 1H, H-6, J_6,5= 7.4Hz),
7.56 (br s, 1H, amide, exchanged with D₂O),
7.34-7.24 (m, 5H, aromatic),
6.31 (d, 1H, H-1 ', J_{1 ', 2'}= 7.3Hz),
5.88 (br s, 1H, H-5), 4.44 (s, 2H, PhCH₂),
4.15 (br s, 1H, H-4 '), 4.06 (d, 1H, H-2', J_{2 ', 1'}= 7.3Hz),
3.97 (d, 1H, H-5'a, J_{5'a, 5'b}= 11.8Hz),
3.82 (dd, 1H, H-5'b, J_{5'b, 4 '} = 1.9, J_{5'b, 5'a}= 11.8Hz),
2.65, 2.59 (each d, each 1H, H-3``a, H-3''b, J = 15.1Hz),
0.95, 0.82 (each s, each 9H, t-Bu),
0.16, 0.15, -0.04, -0.06 (each s, each 3H, Me)
[0114]
Example 22
Synthesis of 1- [3-C- (N-methylcarbamoylmethyl) -β-D-ribofuranosyl] cytosine (Compound 22):
108 mg (0.2 mmol) of the compound 18 obtained in Example 18 was dissolved in 5 ml of methanol, 74 mg (4 mmol) of ammonium fluoride was added, and the mixture was heated to reflux for 2 hours. The reaction mixture was evaporated under reduced pressure, the residue was partitioned by adding 10 ml of water and 10 ml of chloroform, and the aqueous layer was washed with chloroform (2 × 10 ml), concentrated to 10 ml under reduced pressure, and UV absorbed into the supernatant. Activated charcoal was added until there was no more, and it was packed in a column, washed with 50 ml of water and eluted with 50% aqueous methanol. The eluate was concentrated under reduced pressure to obtain 54 mg (yield 86%) of the title compound 22 as a white powder. A white powdery crystal was obtained by crystallization from water / methanol as a sample for analysis.
[0115]
FAB-MS: m / z 315 (MH<sup>+</sup>)
<sup>1</sup>H-NMR (DMSO-d<sub>6</sub>) δ: 8.05 (d, 1H, H-6, J<sub>6,5</sub>= 8.1Hz),
    7.90 (q, 1H, amide, J = 4.5Hz, exchanged with D<sub>2</sub>O),
7.28 (d, 2H, 4-NH<sub>2</sub>, exchanged with D<sub>2</sub>O),
5.91 (d, 1H, H-1 ', J<sub>1 ', 2'</sub>= 8.0Hz),
5.68 (d, 1H, H-5, J<sub>5,6</sub>= 8.1Hz),
5.58 (d, 1H, 2'-OH, J<sub>2'-OH, 2 '</sub> = 6.0Hz, exchanged with D<sub>2</sub>O),
5.25 (t, 1H, 5'-OH, J<sub>5'-OH, 5 '</sub> = 3.9Hz, exchanged with D<sub>2</sub>O),
5.10 (s, 1H, 3'-OH, exchanged with D<sub>2</sub>O),
3.96-3.93 (m, 2H, H-2 ', 4'),
3.75 (ddd, 1H, H-5'a, J<sub>5'a, 4 '</sub> = 1.5, J<sub>5, a, 5'b</sub>= 12.1, J<sub>5'a, 5'-OH</sub>
= 3.9Hz),
3.59 (ddd, 1H, H-5'b, J<sub>5'b, 4 '</sub> = 3.5, J<sub>5'b, 5'a</sub>=
12.1, J<sub>5'b, 5'-OH</sub>= 3.9Hz),
2.55, 2.54 (each d, each 1H, H-3``a, 3''b, J = 16.2Hz).
[0116]
Example 23
Synthesis of 1- [3-C- (N, N-dimethylcarbamoylmethyl) -β-D-ribofuranosyl] cytosine (Compound 23):
In the same manner as in Example 22 from 111 mg (0.2 mmol) of Compound 19 obtained in Example 19, 60.5 mg (yield 92%) of the title compound 23 was obtained as a white powder.
[0117]
FAB-MS: m / z 329 (MH⁺)
¹H-NMR (DMSO-d₆) δ: 7.96 (d, 1H, H-6, J_6,5= 7.3Hz),
7.26 (br d, 2H, 4-NH₂, exchanged with D₂O),
5.91 (d, 1H, H-1 ', J_{1 ', 2'}= 8.1Hz),
5.75 (d, 1H, H-5, J_5,6= 7.3Hz),
5.27 (t, 1H, 5'-OH, J_{5'-OH, 5 '} = 3.9Hz, exchanged with D₂O),
5.26 (d, 1H, 2'-OH, J_{2'-OH, 2 '} = 5.4Hz, exchanged with D₂O),
5.17 (s, 1H, 3'-OH, exchanged with D₂O),
4.03 (br s, 1H, H-4 '),
    3.97 (dd, 1H, H-2 ', J_{2 ', 2'-OH} = 5.4, J_{2 ', 1'}= 7.3Hz),
3.58 (br s, 2H, H-5'a, b), 2.99, 2.85 (each d, each 3H, Me),
2.80, 2.70 (each d, each 1H, H-3``a, 3''b, J = 15.3Hz)
[0118]
Example 24
Synthesis of 1- [3-C- (N-ethylcarbamoylmethyl) -β-D-ribofuranosyl] cytosine (Compound 24):
56.9 mg (yield 86%) of the title compound 24 was obtained as a white powder from 111 mg (0.2 mmol) of the compound 20 obtained in Example 20 in the same manner as in Example 22.
[0119]
FAB-MS: m / z 329 (MH<sup>+</sup>)
<sup>1</sup>H-NMR (DMSO-d<sub>6</sub>) δ: 8.32 (d, 1H, H-6, J<sub>6,5</sub>= 7.8Hz),
8.03 (t, 1H, amide, J<sub>NH, CH2</sub> = 5.4Hz, exchanged with D<sub>2</sub>O),
6.95 (br s, 2H, 4-NH<sub>2</sub>, exchanged with D<sub>2</sub>O),
6.08 (d, 1H, H-1 ', J<sub>1 ', 2'</sub>= 7.8Hz),
5.93 (d, 1H, H-5, J<sub>5,6</sub>= 7.8Hz),
5.76 (d, 1H, 2'-OH, J<sub>2'-OH, 2 '</sub> = 6.5Hz, exchanged with D<sub>2</sub>O),
5.33 (t, 1H, 5'-OH, J<sub>5'-OH, 5 '</sub> = 3.2Hz, exchanged with D<sub>2</sub>O),
5.21 (s, 1H, 3'-OH, exchanged with D<sub>2</sub>O),
4.56 (dq, 2H, CH<sub>2</sub>CH<sub>Three</sub>, J<sub>CH2, NH</sub> = 5.4, J = 7.3Hz),
4.02 (br s, 1H, H-4 '),
3.96 (dd, 1H, H-2 ', J<sub>2 ', 2'-OH</sub> = 6.5, J<sub>2 ', 1'</sub>= 7.8Hz),
3.77 (ddd, 1H, H-5'a, J<sub>5'a, 4 '</sub> = 1.3, J<sub>5'a, 5'b</sub>=
12.5, J<sub>5'a, 5'-OH</sub>= 3.2Hz),
3.61 (ddd, 1H, H-5'b, J<sub>5'b, 4 '</sub> = 3.2, J<sub>5'b, 5'a</sub>=
12.5, J<sub>5'b, 5'-OH</sub>= 3.2Hz),
2.55, 2.54 (each d, each 1H, H-3``a, 3''b, J = 15.3Hz),
1.01 (t, 3H, CH_Three , J = 7.3Hz).
[0120]
Example 25
Synthesis of 1- [3-C- (N-benzylcarbamoylmethyl) -β-D-ribofuranosyl] cytosine (Compound 25):
55.4 mg (yield 71%) of the title compound 25 was obtained as a white powder from 124 mg (0.2 mmol) of the compound 21 obtained in Example 21 in the same manner as in Example 22.
[0121]
FAB-MS: m / z 391 (MH⁺)
¹H-NMR (DMSO-d₆) δ: 8.45 (br t, 1H, amide, J_{NH, CH2} = 5.8Hz,
exchanged with D₂O), 7.91 (d, 1H, H-6, J_6,5= 7.4Hz),
7.33-7.22 (m, 5H, aromatic),
7.16 (br s, 2H, 4-NH₂ , exchanged with D₂O),
5.99 (d, 1H, H-1 ', J_{1 ', 2'}= 7.9Hz),
5.78 (br s, 1H, H-5),
5.42 (d, 1H, 2'-OH, J_{2'-OH, 2 '}= 6.1Hz, exchanged with D₂O),
5.20 (t, 1H, 5'-OH, J_{5'-OH, 5 '} = 4.4Hz, exchanged with D₂O),
5.02 (s, 1H, 3'-OH, exchanged with D₂O),
4.30 (d, 2H, PhCH₂, J_{CH2, NH} = 5.8Hz),
4.02 (dd, 1H, H-2 ', J_{2 ', 2'-OH} = 6.1, J_{2 ', 1'}= 7.9Hz),
3.96 (br s, 1H, H-4 '),
3.70 (ddd, 1H, H-5'a, J_{5'a, 4 '} = 1.2, J_{5'a, 5'b}=
11.9, J_{5'a, 5'-OH}= 4.4Hz),
3.58 (ddd, 1H, H-5'b, J_{5'b, 4 '} = 3.1, J_{5'b, 5'a}=
11.9, J_{5'a, 5'-OH}= 4.4Hz),
2.65, 2.61 (each d, each 1H, H-3``a, 3''b, J = 15.1Hz)
[0122]
Example 26
Synthesis of 1- (2,5-O-di-tert-butyldimethylsilyl-3-C-carbamoylmethyl-β-D-ribofuranosyl) -4-N-methylcytosine (Compound 26):
265 mg (0.5 mmol) of Compound 2 obtained in Example 2 was dissolved in 20 ml of acetonitrile under an argon atmosphere, 2,4,6-triisopropylbenzenesulfonyl chloride 450 mg (1.5 mmol), dimethylaminopyridine 182 mg (1.5 mmol). After adding 0.105 ml (1.5 mmol) of triethylamine and stirring at room temperature for 24 hours, 5 ml of 50% aqueous methylamine solution was added, and the mixture was further stirred at room temperature for 2 hours. The reaction mixture was evaporated under reduced pressure, and the residue was partitioned between chloroform (50 ml) and water (20 ml). The chloroform layer was further washed with water (2 × 20 ml) and saturated brine (20 ml), and dried over anhydrous sodium sulfate. After filtration, the filtrate was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (2 × 15 cm, 5% methanol / chloroform) to obtain 256 mg (yield 94%) of the title compound 26 as yellow crystals. As a sample for analysis, recrystallization from chloroform gave white flat crystals.
[0123]
FAB-MS: m / z 543 (MH<sup>+</sup>)
<sup>1</sup>H-NMR (CDCl<sub>Three</sub>) δ: 8.13 (br s, 1H, amide a, exchanged with D<sub>2</sub>O),
7.79 (d, 1H, H-6, J<sub>6,5</sub>= 8.1Hz),
7.05 (br s, 1H, 4-NH, exchanged with D<sub>2</sub>O),
6.24 (d, 1H, H-1 ', J<sub>1 ', 2'</sub>= 7.2Hz), 5.71 (br s, 1H, H-5),
5.47 (br s, 1H, amide b, exchanged with D<sub>2</sub>O),
4.21 (br s, 1H, H-4 '), 4.10 (d, 1H, H-2', J<sub>2 ', 1'</sub>= 7.2Hz),
3.86 (br s, 2H, H-5'a, b), 2.81 (d, 3H, Me),
2.60 (t, 2H, H-3``a, H-3''b, J = 15.8Hz),
0.93, 0.86 (each s, each 9H, t-Bu),
0.18, 0.17, -0.01, -0.10 (each s, each 3H, Me).
[0124]
Example 27
Synthesis of 1- (2,5-O-di-tert-butyldimethylsilyl-3-C-carbamoylmethyl-β-D-ribofuranosyl) -4-N, N-dimethylcytosine (Compound 27):
In the same manner as in Example 26, 167 mg (0.3 mmol) of Compound 2 obtained in Example 2 and 5 ml of 50% dimethylamine aqueous solution were used to obtain 240 mg (yield 86%) of the title compound 27 as a yellow glassy substance.
[0125]
FAB-MS: m / z 543 (MH⁺)
¹H-NMR (CDCl_Three) δ: 7.99 (d, 1H, H-6, J_6,5= 7.9Hz),
7.10 (br s, 1H, amide a, exchanged with D₂O),
6.27 (d, 1H, H-1 ', J_{1 ', 2'}= 6.7Hz),
5.82 (d, 1H, H-5, J = 7.6Hz),
5.39 (br s, 1H, amide b, exchanged with D₂O),
4.18 (br s, 1H, H-4 '), 4.08 (d, 1H, H-2', J_{2 ', 1'}= 6.7Hz),
3.38 (dd, 1H, H-5'a, J_{5'a, 4 '} = 2.9, J_{5'a, 5'b}= 12.2Hz),
3.28, 3.12 (each br s, each 3H, Me),
2.62, 2.54 (each d, each 1H, H-3``a, H-3''b, J = 15.7Hz),
0.97, 0.86 (each s, each 9H, t-Bu),
0.17, 0.16, -0.00, -0.05 (each s, each 3H, Me).
[0126]
Example 28
Synthesis of 1- (3-C-carbamoylmethyl-β-D-ribofuranosyl) -4-N-methylcytosine (Compound 28):
108 mg (0.2 mmol) of the compound 26 obtained in Example 26 was dissolved in 5 ml of methanol, 74 mg (4 mmol) of ammonium fluoride was added, and the mixture was heated to reflux for 2 hours. The reaction mixture was evaporated under reduced pressure, the residue was partitioned by adding 10 ml of water and 10 ml of chloroform, and the aqueous layer was washed with chloroform (2 × 10 ml), concentrated to 10 ml under reduced pressure, and UV absorbed into the supernatant. Activated charcoal was added until there was no more, and it was packed in a column, washed with 50 ml of water and eluted with 50% aqueous methanol. The eluate was concentrated under reduced pressure to obtain 50 mg (yield 80%) of the title compound 28 as a white powdery substance. A white powdery crystal was obtained by crystallization from water / methanol as a sample for analysis.
[0127]
FAB-MS: m / z 315 (MH<sup>+</sup>)
<sup>1</sup>H-NMR (DMSO-d<sub>6</sub>) δ: 7.96 (d, 1H, H-6, J<sub>6,5</sub>= 7.4Hz),
7.24 (br d, 1H, 4-NH, exchanged with D<sub>2</sub>O),
7.28 (d, 2H, amide, exchanged with D<sub>2</sub>O),
    5.93 (d, 1H, H-5, J<sub>5,6</sub>= 7.9Hz),
5.78 (d, 1H, H-1 ', J<sub>1 ', 2'</sub>= 7.4Hz),
5.50 (d, 1H, 2'-OH, J<sub>2'-OH, 2 '</sub> = 6.0Hz, exchanged with D<sub>2</sub>O),
5.32 (t, 1H, 5'-OH, J<sub>5'-OH, 5 '</sub> = 4.2Hz, exchanged with D<sub>2</sub>O),
5.10 (s, 1H, 3'-OH, exchanged with D<sub>2</sub>O),
4.03 (t, 2H, H-2 ', J<sub>2 ', 1'</sub>= 7.4, J<sub>2 ', 2'-OH</sub> = 4.2Hz),
3.97 (br s, 1H, H-4 '),
3.74 (ddd, 1H, H-5'a, J<sub>5'a, 4 '</sub> = 3.0, J<sub>5'a, 5'b</sub>=
11.9, J<sub>5'a, 5'-OH</sub>= 4.2Hz),
3.62 (ddd, 1H, H-5'b, J<sub>5'b, 4 '</sub> = 3.2, J<sub>5'b, 5'a</sub>=
11.9, J<sub>5'b, 5'-OH</sub>= 4.2Hz), 2.63 (d, 3H, Me, J<sub>Me, NH</sub>= 4.5Hz),
2.57 (s, 2H, H-3``a, 3''b)_.
[0128]
Pharmacological test example 1 (cytocidal action)
1 x 10 human KB cells^FiveIt seed | inoculated to 96-well plate by cells / well. The compound of the present invention was dissolved in purified water, diluted with RPMI 1640 medium to various concentrations, added to each well, and cultured. 5% CO₂After contact at 37 ° C. for 3 days in an incubator, the number of cells was counted by the MTT method.
The cytotoxicity of each compound is determined by the drug concentration that reduces the number of control cells by 50% (IC₅₀). The results are shown in Table 1.
[0129]
[Table 1]

[0130]
As is apparent from the results, the compound of the present invention showed extremely strong cell killing activity as compared with 1- (3-C-ethyl-β-D-ribofuranosyl) uracil which is a known compound.
[0131]
Pharmacological test example 2 (cell killing effect)
Using the cancer cells shown in Table 2, in the same manner as in Pharmacological Test Example 1, compound 7 IC₅₀Asked. The results are shown in Table 2.
[0132]
[Table 2]

[0133]
Formulation examples using the compound of the present invention are shown below.
Formulation Example 1 (Capsule)
Capsules were prepared according to a conventional method at the following blending ratio.
[Table 3]
Compound 3 200mg
Lactose 30mg
Corn starch 50mg
Crystalline cellulose 10mg
Magnesium stearate 3 mg
293mg per capsule
[0134]
Formulation Example 2 (tablet)
Tablets were prepared according to a conventional method at the following blending ratio.
[Table 4]
Compound 7 100mg
Lactose 47mg
Corn starch 50mg
Crystalline cellulose 50mg
Hydroxypropylcellulose 15mg
Talc 2mg
Magnesium stearate 2mg
Ethylcellulose 30mg
Unsaturated fatty acid glycerides 2mg
Titanium dioxide 2 mg
300mg per tablet
[0135]
Formulation Example 3 (Granule)
Granules were prepared according to a conventional method at the following blending ratio.
[Table 5]
Compound 24 200mg
Mannitol 540mg
Corn starch 100mg
Crystalline cellulose 100mg
Hydroxypropylcellulose 50mg
Talc 10 mg
1000mg per package
[0136]
Formulation Example 4 (Fine granules)
A fine granule was prepared according to a conventional method at the following blending ratio.
[Table 6]
Compound 28 200mg
Mannitol 520mg
Corn starch 100mg
Crystalline cellulose 100mg
Hydroxypropylcellulose 70mg
Talc 10 mg
1000mg per package
[0137]
Formulation Example 5 (Injection)
An injection was prepared according to a conventional method at the following blending ratio.
[Table 7]
Compound 7 100mg
Distilled water for injection
2ml in one tube
[0138]
Formulation Example 6 (suppository)
A suppository was prepared according to a conventional method at the following blending ratio.
[Table 8]
Compound 6 200mg
Witepsol S-55
(Saturated fatty acids from lauric acid to stearic acid
Mono-, di-, tri-glyceride mixtures of
Dynamite Nobel) 1300 mg
1500mg per piece

Claims (10)

The following general formula (1)

[ Wherein B represents a nucleobase of cytosine, uracil, 4-N-methylcytosine or 4-N, N-dimethylcytosine , and Z represents a general formula —CH ₂ CONR ⁴ R ⁵ (R ⁴ and R ⁵ are the same) or different from each other, a hydrogen atom, a group represented by a straight or branched alkyl group or a base Njiru group having 1 to 6 carbon atoms), R ^1, R ² and R ³ are the same or different Differently, hydrogen atom, general formula (2)

(Wherein R ^a , R ^b and R ^c are the same or different and represent a linear or branched alkyl group having 1 to 6 carbon atoms or a phenyl group) or a substituted silyl group represented by easily 3'carbamoylalkyl nucleoside derivative or a pharmaceutically acceptable salt thereof represented by shown to] desorbed capable ester forming group in the body. Z is carbamoylmethyl, methylcarbamoylmethyl, diethylcarbamoylmethyl, claim 1 Symbol mounting No 3'carbamoylalkyl nucleoside derivative or a pharmaceutically acceptable salt thereof is Benzylcarbamoyl methyl or dimethylcarbamoylmethyl group. The 3'-carbamoylalkyl nucleoside derivative or the pharmaceutically acceptable salt thereof according to claim 1 or 2 , wherein Z is a carbamoylmethyl group. The 3′-carbamoylalkyl nucleoside derivative or the pharmaceutical thereof according to any one of claims 1 to 3 , wherein the ester-forming residue which can be easily eliminated in vivo represented by R ¹ , R ² and R ³ is an acyl group. Acceptable salt. R < ¹ >, R < ² > and R < ³ > are hydrogen atoms, The 3'-carbamoyl alkyl nucleoside derivative or pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 . B is cytosine, uracil , 4 -N-methylcytosine or 4-N, N-dimethylcytosine, Z is a carbamoylmethyl, methylcarbamoylmethyl, ethylcarbamoylmethyl, benzylcarbamoylmethyl or dimethylcarbamoylmethyl group, R ¹ The 3'-carbamoylalkyl nucleoside derivative according to claim 1, wherein R ² and R ³ are hydrogen atoms, or a pharmaceutically acceptable salt thereof. The B according to claim 1, wherein B is cytosine, uracil, 4-N-methylcytosine or 4-N, N-dimethylcytosine, Z is a carbamoylmethyl group, and R ¹ , R ² and R ³ are hydrogen atoms. 3′-carbamoylalkyl nucleoside derivative or a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising the 3'-carbamoylalkyl nucleoside derivative according to any one of claims 1 to 7 , or a pharmaceutically acceptable salt thereof, and a pharmaceutical carrier. A medicament comprising the 3'-carbamoylalkyl nucleoside derivative or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 7 as an active ingredient. The medicament according to claim 9 , which is an antitumor agent.