JP2019533697A - 5−ハロウラシル修飾マイクロrna及びがんの処置におけるその使用 - Google Patents
5−ハロウラシル修飾マイクロrna及びがんの処置におけるその使用 Download PDFInfo
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Abstract
Description
本出願は、2017年2月28日出願の米国仮特許出願第62/464,491号;2016年11月15日出願の米国仮特許出願第62/422,298号及び2016年11月1日出願の米国仮特許出願第62/415,740号の優先権を主張し、その全体の内容が参照により本明細書に組み込まれる。
本開示は、National Institutes of Healthによって授与された許可番号HL127522及びCA197098下の政府支援によりなされた。政府は、本開示において一定の権利を有する。
2017年10月30日に作成され、United States Patent and Trademark OfficeにEFS-Webを介して提出された、R8859_PCT_SequenceListing.txtと命名された、7KBのASCIIテキストファイル中の配列表は、参照により本明細書に組み込まれる。
用語「マイクロRNA」又は「miRNA」又は「miR」は、互換的に使用され、メッセンジャーRNA分子(mRNA)、DNA又はタンパク質との相互作用により遺伝子の発現を制御する能力があるスモール非コードリボース核酸(RNA)分子を指す。典型的には、マイクロRNAは、約19〜25ヌクレオチド(塩基)の核酸配列から構成され、哺乳動物細胞において見出される。
別の態様において、本開示は、本明細書に記載される修飾核酸組成物の製剤を対象とする。例えば、本核酸組成物は、薬学的用途のため製剤化されうる。特定の実施形態において、製剤は、本明細書に記載される核酸組成物及び薬学的に許容される担体を含有する医薬組成物である。他の実施形態において、本開示の製剤は、修飾miR-129核酸、修飾miR-15a核酸、修飾miR-140核酸、修飾miR-192核酸、修飾miR-502、修飾miR-506核酸又はその組合せ及び薬学的に許容される担体を含む。より具体的には、以下のヌクレオチド配列に示される修飾マイクロRNA核酸は、薬学的適用及び使用のため製剤化されうる;CUFUFUFUFUFGCGGUFCUFGGGCUFUFGC[配列番号4]、CUUUUUGCGGUFCUFGGGCUFUFGC[配列番号5]、UFAGCAGCACAUFAAUFGGUFUFUFGUFG[配列番号6]、UAGCAGCACAUFAAUFGGUFUFUFGUFG[配列番号7]、CAGUFGGUUUUACCCUFAUGGUFAG[配列番号9]、CUFGACCUFAUFGAAUFUFGACAGCC[配列番号1]、AUFCCUFUFGCUAUFCUFGGGUFGCUFA[配列番号13]、及びUFAUFUFCAGGAAGGUFGUFUFACUFUFAA[配列番号15]。
上述したとおり、本開示の修飾マイクロRNA核酸組成物及びその製剤は、天然マイクロRNA及び/又は公知のがん治療(化学療法)、例えば、5-FUによって示される活性と比較した場合に、予想外の及び特別な抗がん活性を示す。したがって、本開示の別の態様は、哺乳動物に本開示の有効量の1つ以上の修飾マイクロRNA核酸組成物又はその製剤を投与する工程による、哺乳動物におけるがんを治療するための方法を提供する。
材料及び方法。
修飾miR-129:5-FU修飾miR-129分子を、自動化オリゴヌクレオチド合成プロセスによって合成し、HPLCによって精製した。2つの鎖をアニールして、成熟修飾5-FU-miR-129を作った。より具体的には、「2'-ACE RNA合成」と呼ばれるプロセスを使用した。2'-ACE RNA合成は、シリルエーテルを用いて、2'-ヒドロキシにおける酸不安定性オルトエステル保護基(2'-ACE)を組み合わせて5'-ヒドロキシル基を保護する、保護基スキームに基づく。この保護基の組合せは、次に標準的なホスホラミダイト固相合成技術で使用される。例えば、それらの各々の全体の内容が本明細書に明示的に組み込まれる、S.A. Scaringe、F.E. Wincott、and M.H. Caruthers、J. Am. Chem. Soc.、120 (45)、11820〜11821 (1998);国際PCT出願WO/1996/041809;M.D. Matteucci、M.H. Caruthers、J. Am. Chem. Soc.、103、3185〜3191 (1981);S.L. Beaucage、M.H. Caruthers、Tetrahedron Lett. 22、1859〜1862 (1981)を参照されたい。
本開示の修飾マイクロRNAは抗がん活性を有する。
図3、図8B、図12A-B、図13A-B及び図14A-Dに示されるように、修飾miRNA(修飾miR:129、15a、192(215)、140、502、及び506)は、非修飾miRNA前駆体よりも、結腸がん、膵がん及び肺がん細胞増殖の阻害においてより有効である。加えて、修飾miRNAは、トランスフェクション試薬なしでがん細胞に送達することができる(データ示さず)。とりわけ、結果は、いくつかの異なる結腸直腸がん細胞株、膵臓がん細胞株、及び肺がん細胞株を越え、がん細胞増殖が、対照マイクロRNAで処置したがん細胞と比較した場合に、有意に阻害されることを示している。
修飾miR-129核酸は抗がん活性を有する。
以下の実験では、5-FUを、miR-129に組み込んだ。1つの実験では、miR-129中の全てのU塩基を、図1Aに提供される構造に示されるように、5-FUで置き換えた、ここで、「UF」は、5-フルオロウラシル又は他の5-ハロウラシルを表す。別の実験では、miR-129のシード領域を除く、全てのU塩基を、図1Bに提供される構造に示されるように、5-FUで置き換えた。
修飾miR-15a及びその抗がん活性。
例示的な修飾miR-15a組成物は、抗がん活性を有する。図1C及び図1Dに示されるように、miR-15a核酸配列の全てのウラシル塩基(図1C)又は非シード領域中のウラシル塩基のみ(図1D)が、5-ハロウラシル(すなわち、5-フルオロウラシル)で置き換えられた、例示的な修飾miR-15aを、上記に示されるように合成した。
Claims (22)
- 少なくとも1つのウラシル核酸を含む修飾マイクロRNAヌクレオチド配列を含む核酸組成物であって、前記少なくとも1つのウラシル核酸の1つ以上が5-ハロウラシルである、核酸組成物。
- 前記修飾マイクロRNAヌクレオチド配列が、miR-129、miR-15a、miR-140、miR-192、miR-502及びmiR-506からなる群から選択されるマイクロRNAヌクレオチド配列を含む、請求項1に記載の核酸組成物。
- 前記修飾マイクロRNAヌクレオチド配列が、CUFUFUFUFUFGCGGUFCUFGGGCUFUFGC[配列番号4]、CUUUUUGCGGUFCUFGGGCUFUFGC[配列番号5]、UFAGCAGCACAUFAAUFGGUFUFUFGUFG[配列番号6]、UAGCAGCACAUFAAUFGGUFUFUFGUFG[配列番号7]、CAGUFGGUUUUACCCUFAUGGUFAG[配列番号9]、CUFGACCUFAUFGAAUFUFGACAGCC[配列番号11]、AUFCCUFUFGCUAUFCUFGGGUFGCUFA[配列番号13]、及びUFAUFUFCAGGAAGGUFGUFUFACUFUFAA[配列番号15]からなる群から選択されるマイクロRNAヌクレオチド配列を含む、請求項2に記載の核酸組成物。
- 前記修飾マイクロRNAヌクレオチド配列が、配列番号1に示されるmiR-129又は配列番号2に示されるmiR-15aのマイクロRNAヌクレオチド配列を含む、請求項2に記載の核酸組成物。
- 前記マイクロRNAヌクレオチド配列が、配列番号1に示されるmiR-129ヌクレオチド配列であり、少なくとも1つのウラシル核酸が5-ハロウラシルである、請求項4に記載の核酸組成物。
- 前記マイクロRNAヌクレオチド配列が、配列番号2に示されるmiR-15aヌクレオチド配列であり、少なくとも1つのウラシル核酸が5-ハロウラシルである、請求項4に記載の核酸組成物。
- 前記5-ハロウラシルが、5-フルオロウラシルである、請求項1に記載の組成物。
- ヌクレオチド配列中のウラシル核酸の少なくとも2つが、5-ハロウラシルである、請求項1に記載の組成物。
- ヌクレオチド配列中のウラシル核酸の少なくとも3つが、5-ハロウラシルである、請求項8に記載の組成物。
- ヌクレオチド配列中のウラシル核酸の少なくとも4つが、5-ハロウラシルである、請求項9に記載の組成物。
- ヌクレオチド配列中のウラシル核酸の少なくとも5つが、5-ハロウラシルである、請求項10に記載の組成物。
- ヌクレオチド配列中のウラシル核酸の少なくとも6つが、5-ハロウラシルである、請求項11に記載の組成物。
- ヌクレオチド配列中のウラシル核酸の全てが、5-ハロウラシルである、請求項1に記載の組成物。
- 前記5-ハロウラシルが、5-フルオロウラシルである、請求項2に記載の組成物。
- 請求項1に記載の核酸組成物及び薬学的に許容される担体を含む医薬組成物。
- 核酸組成物が、CUFUFUFUFUFGCGGUFCUFGGGCUFUFGC[配列番号4]、CUUUUUGCGGUFCUFGGGCUFUFGC[配列番号5]、UFAGCAGCACAUFAAUFGGUFUFUFGUFG[配列番号6]、UAGCAGCACAUFAAUFGGUFUFUFGUFG[配列番号7]、CAGUFGGUUUUACCCUFAUGGUFAG[配列番号9]、CUFGACCUFAUFGAAUFUFGACAGCC[配列番号11]、AUFCCUFUFGCUAUFCUFGGGUFGCUFA[配列番号13]、及びUFAUFUFCAGGAAGGUFGUFUFACUFUFAA[配列番号15]からなる群から選択され、UFが、5-ハロウラシルである、請求項14に記載の医薬組成物。
- 前記5-ハロウラシルが、5-フルオロウラシルである、請求項15に記載の医薬組成物。
- がんを治療するための方法であって、対象に有効量の請求項1に記載の核酸組成物を投与する工程を含み、前記対象が、がんを有するか又はがんを発症する素因を有すると診断されており、前記がんの進行が阻害される、方法。
- 前記哺乳動物がヒトである、請求項18に記載の方法。
- 前記対象が、結腸直腸、膵臓又は肺がんからなる群から選択されるがんを有する、請求項19に記載の方法。
- 前記対象が、結腸直腸がんを有する、請求項20に記載の方法。
- 核酸組成物が、CUFUFUFUFUFGCGGUFCUFGGGCUFUFGC[配列番号4]、CUUUUUGCGGUFCUFGGGCUFUFGC[配列番号5]、UFAGCAGCACAUFAAUFGGUFUFUFGUFG[配列番号6]、UAGCAGCACAUFAAUFGGUFUFUFGUFG[配列番号7]、CAGUFGGUUUUACCCUFAUGGUFAG[配列番号9]、CUFGACCUFAUFGAAUFUFGACAGCC[配列番号11]、AUFCCUFUFGCUAUFCUFGGGUFGCUFA[配列番号13]、及びUFAUFUFCAGGAAGGUFGUFUFACUFUFAA[配列番号15]からなる群から選択され、前記5-ハロウラシルが、5-フルオロウラシルである、請求項17に記載の方法。
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KR20190086688A (ko) | 2019-07-23 |
AU2017353907B2 (en) | 2023-11-30 |
JP7498230B2 (ja) | 2024-06-11 |
CA3042401A1 (en) | 2018-05-11 |
AU2019371830A1 (en) | 2021-06-03 |
MX2019005101A (es) | 2019-08-22 |
EP3873613A4 (en) | 2023-11-15 |
EP3534912A1 (en) | 2019-09-11 |
AU2017353907A1 (en) | 2019-05-23 |
WO2020092466A1 (en) | 2020-05-07 |
IL282836A (en) | 2021-06-30 |
JP2022173194A (ja) | 2022-11-18 |
US20190062754A1 (en) | 2019-02-28 |
KR20210088614A (ko) | 2021-07-14 |
EP3534912A4 (en) | 2020-10-07 |
EP3873613A1 (en) | 2021-09-08 |
WO2018085198A1 (en) | 2018-05-11 |
US11584932B2 (en) | 2023-02-21 |
MX2024000277A (es) | 2024-01-31 |
BR112019008810A2 (pt) | 2019-07-16 |
KR102502248B1 (ko) | 2023-02-21 |
US20230365972A1 (en) | 2023-11-16 |
SG11202104571RA (en) | 2021-06-29 |
CN113573781A (zh) | 2021-10-29 |
CN110290794A (zh) | 2019-09-27 |
JP7130639B2 (ja) | 2022-09-05 |
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