JP2000510446A - Rnaを切断するオリゴリボヌクレオチドおよびリボヌクレアーゼ - Google Patents
Rnaを切断するオリゴリボヌクレオチドおよびリボヌクレアーゼInfo
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Abstract
Description
Claims (1)
- 【特許請求の範囲】 1. あらかじめ選択されたRNA標的に特異的にハイブリダイズすることがで きる合成のオリゴマー性化合物であって、 あらかじめ選択されたRNA標的に対する前記化合物の結合親和性を、RNA標 的に対する修飾されていないオリゴリボヌクレオチドの結合親和性と比較して改 良するよう修飾された少なくとも1つのリボフラノシルヌクレオシドサブユニッ トを含む第1のセグメント;および 2’−ヒドロキシル成分をそこに有する少なくとも4個の連続するリボフラノシ ルヌクレオシドサブユニットを含む第2のセグメント; を含み、 前記オリゴマー性化合物の前記ヌクレオシドサブユニットは、ホスホジエステル 結合と比較して前記結合を分解から安定化するように修飾されているヌクレオチ ド間結合により連結されていることを特徴とする、 合成のオリゴマー性化合物。 2. あらかじめ選択されたRNA標的に対する前記化合物の結合親和性を、R NA標的に対する修飾されていないオリゴリボヌクレオチドの結合親和性と比較 して改良するように修飾されている少なくとも1つのリボフラノシルヌクレオシ ドサブユニットを含む第3のセグメントをさらに含む、請求項1記載のオリゴマ ー性化合物。 3. 前記RNA標的にハイブリダイズしたときに、二本鎖RNAse酵素を活 性化して前記RNA標的を切断することができる、請求項1記載のオリゴマー性 化合物。 4. 前記第2のセグメントが前記第1のセグメントと前記第3のセグメントと の間に位置する、請求項2記載のオリゴマー性化合物。 5. 前記第1のセグメントおよび第3のセグメントがそれぞれ少なくとも3個 のサブユニットを含む、請求項2記載のオリゴマー性化合物。 6. 前記第2のセグメントが4−12個のヌクレオシドサブユニットを含む、 請求項2記載のオリゴマー性化合物。 7. 前記第2のセグメントが5−9個のヌクレオシドサブユニットを含む、請 求項6記載のオリゴマー性化合物。 8. 前記第2のセグメントが少なくとも5個のサブユニットを有し、前記第1 のセグメントおよび第3のセグメントがそれぞれ少なくとも3個のサブユニット を有する、請求項2記載のオリゴマー性化合物。 9. 前記第2のセグメントが少なくとも7個のヌクレオシドサブユニットを有 する、請求項8記載のオリゴマー性化合物。 10. あらかじめ選択されたRNA標的に特異的にハイブリダイズすることが できる合成のオリゴマー性化合物であって、修飾されていない化合物と比較して 、化合物の薬物動態学的結合、吸収、分布またはクリアランス特性;前記化合物 の前記標的RNAに対する親和性または特異性;または前記化合物の電荷の改変 、の少なくとも1つを改良するように修飾されている少なくとも1つのリボフラ ノシルヌクレオシドサブユニットを有する第1のセグメント;および 2’−ヒドロキシル成分をそこに有する少なくとも4個の連続するリボフラノシ ルヌクレオシドサブユニットを含む第2のセグメント; を含み、 前記オリゴマー性化合物の前記ヌクレオシドサブユニットは、ホスホジエステル 結合と比較して前記結合を分解から安定化するように修飾されているヌクレオチ ド間結合により連結されていることを特徴とする、合成のオリゴマー性化合物。 11. 修飾されていない化合物と比較して、化合物の薬物動態学的結合、吸収 、分布またはクリアランス特性;前記化合物の前記標的RNAに対する親和性ま たは特異性;または前記化合物の電荷の改変、の少なくとも1つを改良するよう に修飾されている少なくとも1つのリボフラノシルヌクレオシドサブユニットを 含む第3のセグメントをさらに含む、請求項10記載のオリゴマー性化合物。 12. 前記RNA標的にハイブリダイズしたときに、二本鎖RNAse酵素を 活性化して前記RNA標的を切断しうる、請求項10記載のオリゴマー性化合物 。 13. 前記第2のセグメントが前記第1のセグメントと前記第3のセグメント との間に位置する、請求項11記載のオリゴマー性化合物。 14. 前記第1のセグメントおよび第3のセグメントがそれぞれ少なくとも3 個のサブユニットを含む、請求項11記載のオリゴマー性化合物。 15. 前記第2のセグメントが4−12個のヌクレオシドサブユニットを含む 、請求項11記載のオリゴマー性化合物。 16. 前記第2のセグメントが5−9個のヌクレオシドサブユニットを含む、 請求項15記載のオリゴマー性化合物。 17. 前記第2のセグメントが少なくとも5個のサブユニットを有し、かつ前 記第1のセグメントおよび第3のセグメントがそれぞれ少なくとも3個のサブユ ニットを有する、請求項11記載のオリゴマー性化合物。 18. 前記第2のセグメントが少なくとも7個のヌクレオシドサブユニットを 有する、請求項17記載のオリゴマー性化合物。 19. あらかじめ選択されたRNA標的に特異的にハイブリダイズしうる合成 のオリゴマー性化合物であって、 少なくとも1つの2’−O−C1-20アルキル、2’−O−置換C1-20アルキルま たは2’−フルオロ修飾リボフラノシルヌクレオシドサブユニットを有し、前記 アルキル上の置換が、アミノ、ヒドロキシまたはC1−?アルキルエーテル修飾 である第1のセグメント;および 2’−ヒドロキシル成分をそこに有する少なくとも4個の連続するリボフラノシ ルヌクレオシドサブユニットを含む第2のセグメント; を含み、 前記オリゴマー性化合物の前記ヌクレオシドサブユニットは、ホスホジエステル 結合と比較して分解に対して安定なヌクレオチド間結合により連結されているこ とを特徴とするオリゴマー性化合物。 20. 少なくとも1つの2’−O−C1-20アルキル、2’−O−置換C1-20ア ルキルまたは2’−フルオロ修飾リボフラノシルヌクレオシドサブユニットを含 む第3のセグメントをさらに含み、ここで、前記アルキル上の置換がアミノ、ヒ ドロキシまたはC1-10アルキルエーテルである、請求項19記載のオリゴマー性 化合物。 21. 前記第2のセグメントが前記第1のセグメントと前記第3のセグメント との間に位置する、請求項20記載のオリゴマー性化合物。 22. 前記第1のセグメントおよび第3のセグメントがそれぞれ少なくとも3 個のサブユニットを含む、請求項20記載のオリゴマー性化合物。 23. 前記第2のセグメントが4−12個のヌクレオシドサブユニットを含む 、請求項20記載のオリゴマー性化合物。 24. 前記第2のセグメントが5−9個のヌクレオシドサブユニットを含む、 請求項20記載のオリゴマー性化合物。 25. 前記第2のセグメントが少なくとも5個のサブユニットを有し、および 前記第1のセグメントおよび第3のセグメントがそれぞれ少なくとも3個のサブ ユニットを有する、請求項20記載のオリゴマー性化合物。 26. 前記第2のセグメントが少なくとも7個のヌクレオシドサブユニットを 有する、請求項20記載のオリゴマー性化合物。 27. 前記RNA標的とハイブリダイズしたときに、二本鎖RNAse酵素を 活性化して前記RNA標的を切断しうる、請求項19記載のオリゴマー性化合物 。 28. 前記第1のセグメントおよび前記第3のセグメントの前記リボフラノシ ルヌクレオシドサブユニットがそれぞれ、2’−O−C1-20アルキル、2’−O −置換C1-20アルキルまたは2’−フルオロを有するように修飾されており、か つ前記アルキル上の置換がアミノ、ヒドロキシまたはC1-10アルキルエーテルで ある、請求項22記載のオリゴマー性化合物。 29. 前記ヌクレオシドサブユニットの少なくとも2つが、ホスホロチオエー ト、3’−デオキシ−3’−チオ−ホスホロチオエート、5’−デオキシ−5’ −チオ−ホスホロチオエート、ホスホロジチオエート、ホスホロセレネート、3 ’−デオキシホスフィネート、5’−デオキシホスフィネート、ボラノホスフェ ート、3’−デオキシ−3’−アミノホスホルアミデート、5’−デオキシ−5 ’−アミノホスホルアミデート、水素ホスホネート、ボラノホスフェートエステ ル、ホスホルアミデート、アルキルホスホネート、アリールホスホネートまたは ホスホトリエステル結合により連結されている、請求項19記載のオリゴマー性 化合物。 30. 前記第1のセグメントのヌクレオシドサブユニットの各々が、ホスホロ チオエート、3’−デオキシ−3’−チオ−ホスホロチオエート、5’−デオキ シ−5’−チオ−ホスホロチオエート、ホスホロジチオエート、ホスホロセレネ ート、3’−デオキシホスフィネート、5’−デオキシホスフィネート、ボラノ ホスフェート、3’−デオキシ−3’−アミノホスホルアミデート、5’−デオ キシ−5’−アミノホスホルアミデート、水素ホスホネート、ボラノホスフェー トエステル、ホスホルアミデート、アルキルホスホネート、アリールホスホネー トまたはホスホトリエステル結合により連結されている、請求項19記載のオリ ゴマー性化合物。 31. 前記第1のセグメントの前記ヌクレオシドサブユニットの各々がホスホ ロチオエート結合により連結されている、請求項19記載のオリゴマー性化合物 。 32. 前記第2のセグメントの前記ヌクレオシドサブユニットの各々がホスホ ロチオエート結合により連結されている、請求項19記載のオリゴマー性化合物 。 33. 前記第3のセグメントの前記ヌクレオシドサブユニットの各々が、ホス ホロチオエート、3’−デオキシ−3’−チオ−ホスホロチオエート、5’−デ オキシ−5’−チオ−ホスホロチオエート、ホスホロジチオエート、ホスホロセ レネート、3’−デオキシホスフィネート、5’−デオキシホスフィネート、ボ ラノホスフェート、3’−デオキシ−3’−アミノホスホルアミデート、5’− デオキシ−5’−アミノホスホルアミデート、水素ホスホネート、ボラノホスフ ェートエステル、ホスホルアミデート、アルキルホスホネート、アリールホスホ ネートまたはホスホトリエステル結合により連結されている、請求項20記載の オリゴマー性化合物。 34. 前記第3のセグメントの前記ヌクレオシドサブユニットの各々がホスホ ロチオエート結合により連結されている、請求項20記載のオリゴマー性化合物 。 35. 前記第1のサブユニット、前記第2のサブユニットおよび前記第3のサ ブユニットの前記ヌクレオシドサブユニットがそれぞれ、ホスホロチオエート結 合により連結されている、請求項20記載のオリゴマー性化合物。 36. 前記第1のセグメントのヌクレオシドサブユニットの各々が、カーボネ ート、カルバメート、シリル、イオウ、スルホネート、スルホンアミド、ホルム アセタール、チオホルムアセタール、オキシム、メチレンイミノ、メチレンメチ ルイミノ、メチレンヒドラゾ、メチレンジメチルヒドラゾ、メチレンオキシメチ ルイミノまたはメチレンカルボニルアミノ結合により連結されている、請求項1 9記載のオリゴマー性化合物。 37. 前記第3のセグメントのヌクレオシドサブユニットの各々が、カーボネ ート、カルバメート、シリル、イオウ、スルホネート、スルホンアミド、ホルム アセタール、チオホルムアセタール、オキシム、メチレンイミノ、メチレンメチ ルイミノ、メチレンヒドラゾ、メチレンジメチルヒドラゾ、メチレンオキシメチ ルイミノまたはメチレンカルボニルアミノ結合により連結されている、請求項2 0記載のオリゴマー性化合物。 38. 配列中に少なくとも12個のリボフラノシルヌクレオシドを含み、あら かじめ選択されたRNAに特異的にハイブリダイズしうる合成のオリゴマー性化 合物であって; 前記ヌクレオシドサブユニットは、ホスホジエステル結合と比較して分解に対し てより安定なヌクレオチド間結合により連結されており; 前記化合物はギャップ部分が間に入った2つのウイング部分を有し; ウイング部分はそれぞれ少なくとも1つの修飾ヌクレオシドサブユニットを含み 、前記修飾ヌクレオシドサブユニットは、修飾されていない化合物と比較して、 化合物の薬物動態学的結合、吸収、分布またはクリアランス特性;前記化合物の 前記標的RNAに対する親和性または特異性;または前記化合物の電荷の改変、 の少なくとも1つを改良するように修飾されており、 前記ギャップ部分は、少なくとも4個の連続するリボヌクレオシドサブユニット を有する、 ことを特徴とする化合物。 39. 前記ギャップ部分が少なくとも5個の連続するリボヌクレオシドサブユ ニットを有する、請求項38記載のオリゴマー性化合物。 40. 複数の2’−O−アルキルヌクレオシドサブユニットを有する第1のセ グメント; 少なくとも4個の連続する2’−ヒドロキシルリボヌクレオシドサブユニットを 有する第2のセグメント;および 複数の2’−O−アルキルヌクレオシドサブユニットを有する第3のセグメン ト; を順に含み、オリゴマーのヌクレオシドサブユニットはホスホロチオエートヌク レオチド間結合により連結されている、合成のオリゴマー性化合物。 41. 前記第2のセグメントが少なくとも5個の連続する2’−ヒドロキシル リボヌクレオチドサブユニットを有する、請求項40記載のオリゴマー性化合物 。 42. 前記オリゴマーがあらかじめ選択されたRNAに特異的にハイブリダイ ズしうる、請求項40記載のオリゴマー性化合物。 43. あらかじめ選択されたRNAを特異的に切断する方法であって、 前記RNAを、少なくとも12個のリボフラノシルヌクレオシドサブユニットを 前記あらかじめ選択されたRNAと特異的にハイブリダイズしうる配列で含むオ リゴマー性化合物と接触させることを含み、 前記ヌクレオシドサブユニットは、ホスホジエステル結合と比較して分解に対し てより安定なヌクレオチド間結合により連結されており; 前記化合物は、少なくとも1つの修飾ヌクレオシドサブユニットを含む少なくと も1つのセグメントを有し、前記修飾ヌクレオシドサブユニットは、修飾されて いない化合物と比較して、化合物の薬物動態学的結合、吸収、分布またはクリア ランス特性;前記化合物の前記標的RNAに対する親和性または特異性;または 前記化合物の電荷の改変、の少なくとも1つを改良するように修飾されており; 前記化合物は、少なくとも4個の連続する2’−ヒドロキシルリボヌクレオシド サブユニットを有するさらなるセグメントを有する、 ことを特徴とする方法。 44. 前記さらなるセグメントが少なくとも5個の連続するリボヌクレオシド サブユニットを有する、請求項43記載の方法。 45. 蛋白質の望ましくない産生により特徴づけられる疾患を有する生物を治 療する方法であって、 生物を、リボ核酸の相補鎖と特異的にハイブリダイズしうるヌクレオシドサブユ ニットの配列を有する本発明のオリゴマー性化合物と接触させることを含み、ヌ クレオシドサブユニットの少なくとも1つは、修飾されていない化合物と比較し て、化合物の薬物動態学的結合、吸収、分布またはクリアランス特性;前記標 的RNAに対する前記化合物の親和性または特異性;または前記化合物の電荷の 改変、の少なくとも1つを改良するように修飾されており、 複数のヌクレオシドサブユニットは連続する配列中に位置しており、かつ2’− ヒドロキシル−ペントフラノシル糖成分を有する、 ことを特徴とする方法。 46. 薬学的に有効な量のオリゴマー性化合物を薬学的に許容しうる希釈剤ま たは担体中に含む組成物であって、前記オリゴマー性化合物は、RNAの相補鎖 と特異的にハイブリダイズしうるヌクレオシドサブユニットの配列を含み、オリ ゴマー性化合物の複数のヌクレオシドサブユニットは、修飾されていない化合物 と比較して、化合物の薬物動態学的結合、吸収、分布またはクリアランス特性; 前記標的RNAに対する前記化合物の親和性または特異性;または前記化合物の 電荷の改変、の少なくとも1つを改良するように修飾されており; さらなる複数のヌクレオシドサブユニットが2’−ヒドロキシル−ペントフラノ シル糖成分を有する、 ことを特徴とする組成物。 47. 配列特異的標的RNAのインビトロ修飾の方法であって、dsRNas e酵素および前記標的RNAを含む試験溶液を、前記標的RNAに特異的にハイ ブリダイズしうるヌクレオシドサブユニットの配列を有するオリゴマー性化合物 と接触させることを含み、 ここでヌクレオシドサブユニットの少なくとも1つは、前記化合物の前記標的R NAに対する親和性または特異性を改良するように修飾されており;および 複数のヌクレオシドサブユニットは2’−ヒドロキシル−ペントフラノシル糖成 分を有する、 ことを特徴とする方法。 48. 生物においてハイブリダイゼーションおよびdsRNase酵素活性化 を同時に促進する方法であって、生物を、標的RNAの相補鎖と特異的にハイブ リダイズしうるヌクレオシドサブユニットの配列を有するオリゴマー性化合物と 接触させることを含み、 ここで、ヌクレオシドサブユニットの少なくとも1つは、修飾されていない化合 物と比較して、化合物の薬物動態学的結合、吸収、分布またはクリアランス特性 ;前記化合物の前記標的RNAに対する親和性または特異性;または前記化合物 の電荷の改変;の少なくとも1つを改良するように修飾されており、 複数のヌクレオシドサブユニットは2’−ヒドロキシ−ペントフラノシル糖成分 を有することを特徴とする方法。 49. あらかじめ選択されたRNA標的と特異的にハイブリダイズしうる合成 のオリゴマー性化合物であって; 少なくとも1つの代用物ヌクレオシドサブユニットを有する第1のセグメント; 連続する配列中に位置する少なくとも4個のリボフラノシルヌクレオシドサブユ ニットを含み、かつ2’−ヒドロキシル成分をそこに有する第2のセグメント; を含み、 前記オリゴマー性化合物の前記ヌクレオシドサブユニットは、ホスホジエステル 結合と比較して分解に対して安定なヌクレオチド間結合により連結されているこ とを特徴とする化合物。 50. 前記ヌクレオシドサブユニット代用物がペプチド核酸サブユニットであ る、請求項49記載のオリゴマー性化合物。 51. 前記ヌクレオシドサブユニット代用物がモルホリノヌクレオシドサブユ ニットである、請求項49記載のオリゴマー性化合物。 52. 前記ヌクレオシド代用物がシクロブチルヌクレオシドである、請求項4 9記載のオリゴマー性化合物。 53. 前記ヌクレオシド代用物がピロリジンヌクレオシドである、請求項49 記載のオリゴマー性化合物。 54. あらかじめ選択されたRNA標的と特異的にハイブリダイズしうる合成 のオリゴマー性化合物であって、 少なくとも2つのヌクレオシドサブユニットを含む第1のセグメント;前記第1 のセグメントの前記ヌクレオシドサブユニットは、非リンヌクレオチド間結合に より連結されており;および 2’−ヒドロキシル成分をそこに有する少なくとも4個の連続するリボフラノシ ルヌクレオシドサブユニットを含む第2のセグメント;前記第2のセグメントの 前記ヌクレオシドサブユニットは、ホスホジエステル結合と比較して分解に対し て安定であるヌクレオチド間結合により連結されている、 を含む化合物。 55. 前記非リン結合が、カーボネート、カルバメート、シリル、イオウ、ス ルホネート、スルホンアミド、ホルムアセタール、チオホルムアセタール、オキ シム、メチレンイミノ、メチレンメチルイミノ、メチレン−ヒドラゾ、メチレン ジメチルヒドラゾ、メチレンオキシメチルイミノまたはメチレンカルボニルアミ ノヌクレオチド間結合である、請求項54記載のオリゴマー性化合物。 56. 前記非リンヌクレオチド間結合が、ホルムアセタール、チオホルムアセ タール、メチレンメチルイミノ、メチレンジメチルヒドラゾ、メチレンオキシメ チルイミノまたはメチレンカルボニルアミノヌクレオチド間結合である、請求項 54記載のオリゴマー性化合物。 57. 前記第2のセグメントの前記ヌクレオシドサブユニットがホスホロチオ エートヌクレオチド間結合により連結されている、請求項54記載のオリゴマー 性化合物。 58. あらかじめ選択されたRNA標的に特異的にハイブリダイズしうる合成 のオリゴマー性化合物であって、 少なくとも3個のヌクレオシドサブユニットを含む第1のセグメント;前記第1 のセグメントの前記ヌクレオシドサブユニットは、交互のリンおよび非リンヌク レオチド間結合により連結されており;および 2’−ヒドロキシル成分をそこに有する少なくとも4個の連続するリボフラノシ ルヌクレオシドサブユニットを含む第2のセグメント;前記第2のセグメントの 前記ヌクレオシドサブユニットは、ホスホジエステル結合と比較して分解に対し てより安定なヌクレオチド間結合により連結されている、 を含む化合物。 59. 前記非リン結合が、カーボネート、カルバメート、シリル、イオウ、ス ルホネート、スルホンアミド、ホルムアセタール、チオホルムアセタール、オキ シム、メチレンイミノ、メチレンメチルイミノ、メチレンヒドラゾ、メチレンジ メチルヒドラゾ、メチレンオキシメチルイミノまたはメチレンカルボニルアミノ ヌクレオチド間結合である、請求項58記載のオリゴマー性化合物。 60. 前記非リンヌクレオチド間結合が、ホルムアセタール、チオホルムアセ タール、メチレンメチルイミノ、メチレンジメチルヒドラゾ、メチレンオキシメ チルイミノまたはメチレンカルボニルアミノヌクレオチド間結合である、請求項 58記載のオリゴマー性化合物。 61. 前記第2のセグメントの前記ヌクレオシドサブユニットが、ホスホロチ オエートヌクレオチド間結合により連結されている、請求項58記載のオリゴマ ー性化合物。 62. あらかじめ選択されたRNA標的に特異的にハイブリダイズしうる合成 のオリゴマー性化合物であって; 少なくとも2つのヌクレオシドサブユニットを含む第1のセグメント;前記第1 のセグメントの前記ヌクレオシドサブユニットは、3’−デオキシ−3’−チオ −ホスホロチオエート、5’−デオキシ−5’−チオ−ホスホロチオエート、ホ スホロジチオエート、ホスホロセレネート、3’−デオキシホスフィネート、5 ’−デオキシホスフィネート、ボラノホスフェート、3’−デオキシ−3’−ア ミノホスホルアミデート、5’−デオキシ−5’−アミノホスホルアミデート、 水素ホスホネート、ボラノホスフェートエステル、ホスホルアミデート、アルキ ルホスホネート、アリールホスホネートまたはホスホトリエステルホスフェート 結合により連結されており;および 2’−ヒドロキシル成分をそこに有する少なくとも4個の連続するリボフラノシ ルヌクレオシドサブユニットを含む第2のセグメント;前記第2のセグメントの 前記ヌクレオシドサブユニットは、ホスホジエステル結合と比較して分解に対し てより安定であるヌクレオチド間結合により連結されている、 を含む化合物。 63. 少なくとも2つのヌクレオシドサブユニットの第3のセグメントを含み 、前記第3のセグメントの前記ヌクレオシドサブユニットが、3’−デオキシ− 3’−チオ−ホスホロチオエート、5’−デオキシ−5’−チオ−ホスホロチオ エート、ホスホロジチオエート、ホスホロセレネート、3’−デオキシホスフィ ネート、5’−デオキシホスフィネート、ボラノホスフェート、3’−デオキシ −3’ −アミノホスホルアミデート、5’−デオキシ−5’−アミノホスホルアミデー ト、水素ホスホネート、ボラノホスフェートエステル、ホスホルアミデート、ア ルキルホスホネート、アリールホスホネートまたはホスホトリエステルホスフェ ート結合により連結されている、請求項62記載のオリゴマー性化合物。 64. 前記第1のセグメントおよび第3のセグメントの前記ヌクレオシドサブ ユニットが、ホスホロジチオエート、ホスホロセレネート、3’−デオキシホス フィネート、3’−デオキシ−3’−アミノホスホルアミデート、ホスホルアミ デート、アルキルホスホネート、アリールホスホネートまたはホスホトリエステ ルホスフェート結合により連結されている、請求項62記載のオリゴマー性化合 物。 65. あらかじめ選択されたRNA標的に特異的にハイブリダイズしうる合成 のオリゴマー性化合物であって、 DNAまたはRNA主要構築ブロックヌクレオシドではない少なくとも1つのリ ボフラノシルヌクレオシドサブユニットを有する第1のセグメント; 2’−ヒドロキシル成分をそこに有する少なくとも4個の連続するリボフラノシ ルヌクレオシドサブユニットを含む第2のセグメント; を含み、 前記オリゴマー性化合物の前記ヌクレオシドサブユニットは、ホスホジエステル 結合と比較して前記結合を分解から安定化するように修飾されたヌクレオチド間 結合により連結されている、ことを特徴とする化合物。 66. 前記第1のセグメントヌクレオシドサブユニットが、キサンチン、ヒポ キサンチン、2−アミノアデニン、アデニンおよびグアニンの6−アルキル誘導 体、アデニンおよびグアニンの2−アルキル誘導体、アデニンおよびグアニンの 7−アルキル誘導体、5−ハロウラシルおよびシトシン、5−プロピニルウラシ ルおよびシトシン、6−アゾウラシル、シトシンおよびチミン、5−ウラシル( シュードウラシル)、4−チオウラシル、2−チオウラシルおよびシトシン、8 −ハロ、アミノ、チオール、チオアルキル、ヒドロキシルアデニンおよびグアニ ン、および5−トリフルオロメチルウラシルおよびシトシンをその複素環塩基と して有するヌクレオシドから選択される、請求項65記載の化合物。 67. あらかじめ選択されたRNA標的と特異的にハイブリダイズすることが でき、 アデノシン、2’−デオキシアデノシン、グアノシン、2’−デオキシグアノシ ン、シチジン、2’−デオキシシチジン、ウリジンおよび2’−デオキシチミジ ンからなるヌクレオシド群を除く少なくとも1つのリボフラノシルヌクレオシド サブユニットを有する第1のセグメント; 2’−ヒドロキシル成分をそこに有する少なくとも4個の連続するリボフラノシ ルヌクレオシドサブユニットを含む第2のセグメント; を含む合成のオリゴマー性化合物であって、 前記オリゴマー性化合物の前記ヌクレオシドサブユニットは、ホスホジエステル 結合と比較して前記結合を分解から安定化するように修飾されているヌクレオチ ド間結合により連結されていることを特徴とするオリゴマー性化合物。 68. 二本鎖基質の分解を触媒する活性を有する哺乳動物リボヌクレアーゼで あって、前記基質の前記鎖の一方はmRNAであり、および前記基質の前記鎖の 他方は、複数の2’修飾ヌクレオシドサブユニットを含む第1のセグメントおよ び2’−ヒドロキシル成分をそこに有する少なくとも4個の連続するリボフラノ シルヌクレオシドサブユニットを含む第2のセグメントを順に有する化合物を含 むことを特徴とするリボヌクレアーゼ。 69. 前記化合物の前記サブユニットが、ホスホロチオエートヌクレオチド間 結合またはホスホジエステルヌクレオチド間結合により連結されている、請求項 68記載の哺乳動物リボヌクレアーゼ。 70. 前記化合物の前記第1のセグメントの前記サブユニットが、ホスホロチ オエートヌクレオチド間結合により連結されている、請求項68記載の哺乳動物 リボヌクレアーゼ。 71. 前記化合物の前記第2のセグメントの前記サブユニットが、ホスホジエ ステルヌクレオチド間結合により連結されている、請求項70記載の哺乳動物ボ ヌクレアーゼ。 72. 前記化合物の前記第2のセグメントの前記サブユニットが、ホスホロチ オエートヌクレオチド間結合により連結されている、請求項70記載の哺乳動物 リボヌクレアーゼ。 73. 前記化合物の前記第1のセグメントの前記サブユニットが、2’−O− アルキルヌクレオシドサブユニットである、請求項68記載の哺乳動物リボヌク レアーゼ。 74. (A) 前記活性はNaClにより阻害され; (B) 前記活性はMg++を必要とし;および (D) 前記哺乳動物リボヌクレアーゼは、SDS−PAGEにより決定して約 50から約80キロダルトンの見かけの分子量を有する、 請求項68記載の哺乳動物リボヌクレアーゼ。 75. 前記リボヌクレアーゼが核から単離される、請求項68記載の哺乳動物 リボヌクレアーゼ。 76. 前記リボヌクレアーゼが細胞質から単離される、請求項68記載の哺乳 動物リボヌクレアーゼ。 77. 前記リボヌクレアーゼがヒト細胞または組織から単離しうる、請求項6 8記載の哺乳動物蛋白質。 78. 第1のオリゴヌクレオチドおよび第2のオリゴヌクレオチドのデュープ レックスを含む二本鎖RNA基質であって、 (A) 前記第1のオリゴヌクレオチドおよび前記第2のオリゴヌクレオチドは それぞれホスホジエステル結合を有する少なくとも4個の連続するリボフラノシ ル残基を有する中心部分を有し、 ここで前記中心部分は、前記デュープレックス中で互いに塩基対を形成しており ; (B) 前記第1のオリゴヌクレオチドおよび前記第2のオリゴヌクレオチドの 少なくとも1つは、一本鎖ヌクレアーゼに対してそれらを耐性にする化学修飾を 有する、前記中心部分に隣接する部分を有する ことを特徴とする基質。 79. 第1のオリゴヌクレオチドおよび第2のオリゴヌクレオチドのデュープ レックスを含む二本鎖RNA基質であって、 (A) 前記第1のオリゴヌクレオチドおよび前記第2のオリゴヌクレオチドは それぞれホスホジエステル結合を有する少なくとも4個の連続するリボフラノシ ル残基を有する中心部分を有し、ここで前記中心部分は前記デュープレックス中 において互いに塩基対を形成しており; (B) 前記第1のオリゴヌクレオチドおよび前記第2のオリゴヌクレオチドの 少なくとも1つは、一本鎖ヌクレアーゼに対してそれらを耐性にし、かつデュー プレックスの他方のオリゴヌクレオチドに対するそれらの親和性を増加させる化 学修飾を有する、前記中心部分に隣接する部分を有する ことを特徴とする基質。 80. 前記化学修飾がホスホロチオエート結合または2’−メトキシ修飾であ る、請求項78記載の二本鎖RNA基質。 81. 請求項78記載のdsRNA基質を含む親和性マトリックス。 82. リボヌクレアーゼまたは非分解性RNA結合蛋白質を精製する方法であ って、前記リボヌクレアーゼまたは非分解性RNA結合蛋白質を含む試料を請求 項81記載の親和性マトリックスと接触させることを含む方法。 83. ホスホロチオエートヌクレオチド間結合により連結されている複数の2 ’−O−アルキルヌクレオシドサブユニットを有する第1のセグメント;および ホスホロチオエートヌクレオチド間結合またはホスホジエステルヌクレオチド間 結合により連結されている少なくとも4個の連続する2’−ヒドロキシルリボヌ クレオシドサブユニットを有する第2のセグメント を順に含む、合成のオリゴマー性化合物。 84. ホスホロチオエートヌクレオチド間結合により連結されている複数の2 ’−O−アルキルヌクレオシドサブユニットを有する第3のセグメントをさらに 含み、前記第2のセグメントは、前記オリゴマー性化合物中で前記第1のセグメ ントと前記第3のセグメントとの間に位置している、請求項83記載の化合物。 85. 前記第2のセグメントがホスホジエステルヌクレオチド間結合を有する 、請求項83記載の化合物。 86. 前記第2のセグメントがホスホロチオエートヌクレオチド間結合を有す る、請求項83記載の化合物。 87. ホスホロチオエートヌクレオチド間結合により連結されている複数の2 ’−O−アルキルヌクレオシドサブユニットを有する第1のセグメント; ホスホロチオエートヌクレオチド間結合またはホスホジエステルヌクレオチド間 結合により連結されている少なくとも4個の連続する2’−ヒドロキシルリボヌ クレオシドサブユニットを有する第2のセグメント;および ホスホロチオエートヌクレオチド間結合により連結されている複数の2’−O− アルキルヌクレオシドサブユニットを有する第3のセグメント を順に含む、合成のオリゴマー性化合物 88. 前記第2のセグメントがホスホロチオエートヌクレオチド間結合を有す る、請求項87記載の化合物。 89. 前記mRMAによりコードされる蛋白質の望ましくない産生により特徴 づけられる疾患を有する生物を治療するための、請求項68記載の前記リボヌク レアーゼの使用。 90. 前記mRNAまたは前記mRNAによりコードされる蛋白質の1つを同 定するための、請求項68記載の前記リボヌクレアーゼの使用。 91. 生物において、前記mRNAによりコードされる蛋白質に関連する異常 な状態を診断するための、請求項68記載の前記リボヌクレアーゼの使用。 92. 二本鎖基質の分解を触媒する活性を有する哺乳動物リボヌクレアーゼで あって、前記基質の前記鎖の一方がmRNAであり、かつ前記鎖の他方が請求項 1記載の化合物を含むことを特徴とするリボヌクレアーゼ。 93. 前記オリゴヌクレオチドの1つが配列番号8のヌクレオチド配列を有す る、請求項78記載の二本鎖RNA基質。
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1996
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- 1997-06-06 WO PCT/US1997/009963 patent/WO1997046570A1/en active IP Right Grant
- 1997-06-06 EP EP04078164A patent/EP1621545A3/en not_active Withdrawn
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
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WO2005021571A1 (ja) * | 2003-08-29 | 2005-03-10 | Mitsui Chemicals, Inc. | N4−ベンゾイルシチジン誘導体の製造法 |
JP2010136725A (ja) * | 2003-11-03 | 2010-06-24 | Isis Pharmaceuticals Inc | Sglt2発現の調節 |
JP2016513950A (ja) * | 2012-09-26 | 2016-05-19 | ミルクス セラピューティクス アクティーゼルスカブMirrx Therapeutics A/S | オフターゲットプロファイルの改善されたオリゴマー |
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EP0928290A4 (en) | 2000-01-19 |
EP2361923A2 (en) | 2011-08-31 |
US7695902B2 (en) | 2010-04-13 |
AU3383297A (en) | 1998-01-05 |
US7432250B2 (en) | 2008-10-07 |
ATE292141T1 (de) | 2005-04-15 |
EP1621545A3 (en) | 2008-08-06 |
WO1997046570A1 (en) | 1997-12-11 |
US7919612B2 (en) | 2011-04-05 |
EP0928290B9 (en) | 2005-11-09 |
US7629321B2 (en) | 2009-12-08 |
EP2361923A3 (en) | 2012-10-31 |
EP1600506A2 (en) | 2005-11-30 |
US20030096784A1 (en) | 2003-05-22 |
EP0928290B1 (en) | 2005-03-30 |
DE69732911T2 (de) | 2006-05-11 |
EP1600506A3 (en) | 2008-07-09 |
JP5006485B2 (ja) | 2012-08-22 |
US20040171033A1 (en) | 2004-09-02 |
DE69732911D1 (de) | 2005-05-04 |
US20030096286A1 (en) | 2003-05-22 |
US7432249B2 (en) | 2008-10-07 |
JP2011142911A (ja) | 2011-07-28 |
US6107094A (en) | 2000-08-22 |
EP1621545A2 (en) | 2006-02-01 |
US20100151458A1 (en) | 2010-06-17 |
US5898031A (en) | 1999-04-27 |
US20030096287A1 (en) | 2003-05-22 |
US20040191773A1 (en) | 2004-09-30 |
EP0928290A1 (en) | 1999-07-14 |
US20030119777A1 (en) | 2003-06-26 |
US20020165189A1 (en) | 2002-11-07 |
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