JP2022544457A - 核酸-リン酸カルシウムナノ粒子複合体及び生物鉱化におけるその使用 - Google Patents
核酸-リン酸カルシウムナノ粒子複合体及び生物鉱化におけるその使用 Download PDFInfo
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- JP2022544457A JP2022544457A JP2022505251A JP2022505251A JP2022544457A JP 2022544457 A JP2022544457 A JP 2022544457A JP 2022505251 A JP2022505251 A JP 2022505251A JP 2022505251 A JP2022505251 A JP 2022505251A JP 2022544457 A JP2022544457 A JP 2022544457A
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- biomineralizer
- bone
- nucleic acid
- collagen
- rna
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Abstract
Description
Claims (22)
- 生物鉱化剤であって、核酸と非晶質リン酸カルシウムナノ粒子で形成された複合体を含み、好ましくは、静電吸着により形成される、ことを特徴とする生物鉱化剤。
- 前記複合体において、前記核酸と非晶質リン酸カルシウムナノ粒子は、
核酸のリン酸基とナノ粒子のカルシウムイオンとの静電吸着と、
遊離のリン酸イオンと核酸に吸着したカルシウムイオンとの更なる静電吸着と、によって形成される、ことを特徴とする請求項1に記載の生物鉱化剤。 - 核酸溶液、塩化カルシウム溶液及びリン酸水素二カリウム溶液で混合されて形成される、請求項1~2のいずれか1項に記載の生物鉱化剤。
- カルシウムイオンの濃度は、1~10mMであり、好ましくは、2~5mMであり、より好ましくは、1.67~3.5mMであり、リン酸イオンの濃度は、0.5~10mMであり、好ましくは、1~5mMであり、より好ましくは、1.0~2.1mMである、ことを特徴とする請求項1~3のいずれか1項に記載の生物鉱化剤。
- カルシウムイオンとリン酸イオンの比は、10:1~1:5であり、好ましくは、5:1~1:3であり、好ましくは、3.5:1~1:1.25であり、より好ましくは、2:1~1:1であり、例えば、1.67:1である、ことを特徴とする請求項1~4のいずれか1項に記載の生物鉱化剤。
- 前記複合体の粒径が、1~100nmであり、好ましくは、10~100nmであり、より好ましくは、20~100nmであり、例えば、40~60nm又は60~100nmである、ことを特徴とする請求項1~5のいずれか1項に記載の生物鉱化剤。
- 前記核酸は、哺乳動物細胞から単離された全DNA又は全RNA又はプラスミドDNAなどのDNA又はRNAであり、又は、前記核酸は、miR-17-92、miR-26a又はmiR-148b又はBMP2-プラスミドDNAなどの、骨形成分化を促進し、及び/又は骨再生を促進する核酸を含む、ことを特徴とする請求項1~6のいずれか1項に記載の生物鉱化剤。
- 前記哺乳動物細胞は、骨細胞前駆細胞、前骨芽細胞、骨細胞、骨形成細胞、骨芽細胞、破骨細胞又は骨髄間葉系幹細胞である、ことを特徴とする請求項1~7のいずれか1項に記載の生物鉱化剤。
- 核酸の作用濃度は、10~500ug/mlであり、好ましい作用濃度は、50~500ug/mlであり、より好ましい作用濃度は、100~300ug/mlであり、例えば、150~250ug/mlである、請求項1~8のいずれか1項に記載の生物鉱化剤。
- そのpH値は、5.5~7であり、好ましくは、6.0~6.5である、請求項1~9のいずれか1項に記載の生物鉱化剤。
- 例えば、溶液又はコロイド液のような液体形態、ゲルなどの半固体形態、粉末又は凍結乾燥粉末のような固体形態であってもよい、ことを特徴とする請求項1~10のいずれか1項に記載の生物鉱化剤。
- 請求項1~11のいずれか1項に記載の生物鉱化剤を含み、又は請求項1~11のいずれかに記載の生物鉱化剤で処理される、ことを特徴とする鉱化されたコラーゲン線維製品。
- コラーゲン足場、コラーゲン膜、コラーゲン線維シート、脱灰骨組織、脱灰象牙質スライス、マウス尾部、歯又は骨修復材料、歯又は骨足場材料、歯又は骨の再生材料、又は歯又は骨埋め込み材料から選択される、ことを特徴とする請求項12に記載のコラーゲン線維製品。
- 患者の骨関連疾患又は障害を治療し、又は骨の状態を改善するための医薬又は医療機器の製造における、請求項1~11のいずれか1項に記載の生物鉱化剤又は請求項12~13のいずれか1項に記載の鉱化されたコラーゲン線維製品の使用。
- 前記医薬又は機器は、患者の骨欠損又は骨損失を治療し、又は骨修復、骨分化又は骨再生を促進するために用いられる、ことを特徴とする請求項14に記載の使用。
- 患者のコラーゲン線維を鉱化するための医薬の製造における、請求項1~10のいずれか1項に記載の生物鉱化剤の使用。
- 請求項1~11のいずれか1項に記載の生物鉱化剤を製造するための方法であって、
(1)、好ましくは、哺乳動物細胞から全DNA(各種のDNAを含む)又は全RNA(各種のRNAを含む)を抽出し、又はプラスミドDNAを抽出するための核酸を得るステップと、
(2)、ステップ(1)で得られた核酸を塩化カルシウム溶液と混合するステップと、
(3)、ステップ(2)で得られた混合物にリン酸水素二カリウム溶液を添加することにより、核酸と非晶質リン酸カルシウムナノ粒子とで形成された複合体を得るステップとを含む、ことを特徴とする方法。 - ステップ(1)で取得された核酸の出発濃度は、100~5000ng/μlであり、好ましい出発濃度は、500~5000ng/μlであり、より好ましい出発濃度は、1000~3000ng/μlであり、例えば、1500~2500ng/μlであり,核酸分子量の範囲が広く、好ましくは、核酸分子量が40kDaよりも大きい、ことを特徴とする請求項17に記載の方法。
- コラーゲン線維の生体模倣型鉱化を誘導し、又は鉱化されたコラーゲン線維製品を製造する方法であって、請求項1~11のいずれか1項に記載の生物鉱化剤を、コラーゲン線維又はコラーゲン線維を含む製品と接触させることを含む、ことを特徴とする方法。
- 核酸送達又はトランスフェクションシステムであって、前記核酸送達は、請求項1~10のいずれか1項に記載の生物鉱化剤を含む、ことを特徴とする核酸送達又はトランスフェクションシステム。
- 細胞トランスフェクションに用いることができ、骨欠損における骨形成促進タンパク質の発現を制御及び維持し、又は、骨形成分化及び骨再生を促進するために用いられる、ことを特徴とする請求項20に記載の核酸送達又はトランスフェクションシステム。
- 前記生物鉱化剤が、骨形成分化を促進し、及び/又は骨再生を促進する核酸を含む、ことを特徴とする請求項20又は21に記載の核酸送達又はトランスフェクションシステム。
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