JP7323229B2 - ケロイドまたは肥厚性瘢痕の予防または治療用組成物 - Google Patents
ケロイドまたは肥厚性瘢痕の予防または治療用組成物 Download PDFInfo
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Description
(a)ケロイド組織またはケロイド細胞に試験物質を処理する段階;および
(b)前記試験物質が処理された組織または細胞で細胞内TXNDC5、PRRC1、S100A11、Galectin 1、Filamin A、eIF-5A、Annexin A2およびFABP5よりなる群から選ばれる一つ以上のタンパク質の活性または前記タンパク質をコードする遺伝子の発現量を分析する段階であって、正常組織での前記タンパク質の活性またはこれをコードする遺伝子の発現様相と異なる場合、前記試験物質をケロイドまたは肥厚性瘢痕の予防または治療物質と判断することができる。
本発明の態様を以下の項にさらに記載する:
[項1]
TXNDC5、PRRC1、S100A11、Galectin 1、Filamin A、eIF-5A、Annexin A2およびFABP5よりなる群から選ばれる一つ以上のタンパク質の活性を抑制したり、または前記一つ以上のタンパク質をコードする遺伝子の発現を抑制する物質を有効成分として含む、ケロイド(Keloid)または肥厚性瘢痕(Hypertrophic scar)の予防または治療用薬学的組成物。
[項2]
前記有効成分は、shRNA、siRNA、miRNA、アンチセンスオリゴヌクレオチド、リボザイム(ribozyme)、Crispr-cas9、DNAzyme、PNA(peptide nucleic acids)、ペプチド、抗体、アプタマー、天然抽出物または化学物質であることを特徴とする上記項1に記載のケロイド(Keloid)または肥厚性瘢痕(Hypertrophic scar)の予防または治療用薬学的組成物。
[項3]
前記有効成分は、前記遺伝子の発現を抑制するshRNA、siRNA、miRNAまたはアンチセンスオリゴヌクレオチドであることを特徴とする上記項2に記載のケロイド(Keloid)または肥厚性瘢痕(Hypertrophic scar)の予防または治療用薬学的組成物。
[項4]
(a)ケロイド(Keloid)または肥厚性瘢痕(Hypertrophic scar)患者に由来した組織に試験物質を処理する段階;および
(b)前記試験物質が処理された組織または細胞で TXNDC5、PRRC1、S100A11、Galectin 1、Filamin A、eIF-5A、Annexin A2およびFABP5よりなる群から選ばれる一つ以上のタンパク質の活性または前記タンパク質をコードする遺伝子の発現量を分析する段階であって、正常組織での前記タンパク質の活性またはこれをコードする遺伝子の発現様相と異なる場合、前記試験物質をケロイドまたは肥厚性瘢痕の予防または治療物質と判断する、ケロイド(Keloid)または肥厚性瘢痕(Hypertrophic scar)の予防または治療物質のスクリーニング方法。
[項5]
TXNDC5、PRRC1、S100A11、Galectin 1、Filamin A、eIF-5A、Annexin A2およびFABP5よりなる群から選ばれる一つ以上の遺伝子、またはTXNDC5、PRRC1、S100A11、Galectin 1、Filamin A、eIF-5A、Annexin A2およびFABP5よりなる群から選ばれる一つ以上のタンパク質の発現量を測定する物質を含む、ケロイド(Keloid)または肥厚性瘢痕(Hypertrophic scar)診断用組成物。
[項6]
ケロイドまたは肥厚性瘢痕を有したり予想される個体から得た試料でTXNDC5、PRRC1、S100A11、Galectin 1、Filamin A、eIF-5A、Annexin A2およびFABP5よりなる群から選ばれる一つ以上の遺伝子、またはTXNDC5、PRRC1、S100A11、Galectin 1、Filamin A、eIF-5A、Annexin A2およびFABP5よりなる群から選ばれる一つ以上のタンパク質の発現量を測定する段階;および
前記測定結果を正常個体の試料での前記遺伝子またはタンパク質の発現量と比較する段階;を含むケロイドまたは肥厚性瘢痕の診断方法。
[項7]
ケロイドまたは肥厚性パヌルルル有する個体に(a)有効成分としてTXNDC5、PRRC1、S100A11、Galectin 1、Filamin A、eIF-5A、Annexin A2およびFABP5よりなる群から選ばれる一つ以上の遺伝子の発現を抑制したり、またはTXNDC5、PRRC1、S100A11、Galectin 1、Filamin A、eIF-5A、Annexin A2およびFABP5よりなる群から選ばれる一つ以上のタンパク質の活性を抑制する物質を含む薬学的組成物を投与することを含む、ケロイド疾患または肥厚性瘢痕を有する個体を治療する方法。
ヒト皮膚線維芽細胞、正常組織および肥厚性瘢痕組織の準備
実験に使用した皮膚組織は、総3人の患者から獲得し、男女の正常と瘢痕(scar)組織を利用してIHCを進め、各組織に由来した初代線維芽細胞(primary fibroblast cell)を利用してsiRNA形質転換による効果を観察した。
Biopsyで得られた組織をO.C.T compound(Cell Poth,KMA-0100-00A)を入れ、ドライアイスに入れて凍結した。凍結した組織をアセトンとメタノールが1:1で含有されたバッファーで固定した。バッファーの除去後、0.5%Triton X-100を常温で10分間処理してpermeabilizationさせた後、Ultravision hydrogen peroxide(Thermo kit/Ultravision LP detection system)を利用してペルオキシダーゼ(peroxidase)活性を阻害した。Ultravision blockバッファーを常温で10分間反応させた後、当該1次抗体を処理し、4℃でオーバーナイト処理した。一次抗体は、それぞれTXNDC5(Abcam,Ab155684)、PRRC1(Abcam/ab12544)、S100A11(Abcam/ab97329)、Galectin 1(Abcam,Ab108389)、Filamin A(Millipore/MAB1680)、eIF5-A(Abcam/ab32014)、Annexin A2(Cell singnaling/8235)、FABP5(Abcam/ab37267)を使用した。一次抗体エンハンサー(Primary antibody enhancer)を常温で10分間処理した後、HRP polymerを光遮断状態で常温で15分間反応させた。HRP-polymerを除去した後、PRRC1を除いたすべての組織をAEC(Spring,ASS-125)、PRRC1は、DAB(Thermo,TA-125-HDX)を1分間処理した。Mayer’s hematoxylinで常温で組織を染色し、脱水させた後、Canada balsam mixed Xyleneでmounting進行後、組織を顕微鏡で観察した。
前記IHC結果で選別された前記タンパク質のうち3種(TXNDC5,PRRC1,S100A11)に対してsiRNAを利用して各遺伝子発現をノックダウン(knock-down)させた後、傷跡で過度に発現するコラーゲンおよび線維芽細胞増殖関連タンパク質などの発現を分析した。
Claims (2)
- PRRC1の発現レベルの測定のためのPRRC1に特異的に結合する抗体を含む、肥厚性瘢痕(Hypertrophic scar)診断用組成物。
- TXNDC5、S100A11、Galectin 1、Filamin A、eIF-5A、Annexin A2およびFABP5よりなる群から選ばれる一つ以上のタンパク質の発現レベルの測定のための、TXNDC5、S100A11、Galectin 1、Filamin A、eIF-5A、Annexin A2およびFABP5よりなる群から選ばれる一つ以上のタンパク質に特異的に結合する抗体をさらに含む、請求項1に記載の組成物。
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KR1020190117157A KR102100163B1 (ko) | 2019-09-24 | 2019-09-24 | 켈로이드 또는 비후성 반흔 예방 또는 치료용 조성물 |
KR10-2019-0117157 | 2019-09-24 | ||
PCT/KR2019/017039 WO2021060624A1 (ko) | 2019-09-24 | 2019-12-04 | 켈로이드 또는 비후성 반흔 예방 또는 치료용 조성물 |
JP2020509485A JP7132646B2 (ja) | 2019-09-24 | 2019-12-04 | ケロイドまたは肥厚性瘢痕の予防または治療用組成物 |
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CN112839664A (zh) | 2021-05-25 |
EP3824892A1 (en) | 2021-05-26 |
US20210388352A1 (en) | 2021-12-16 |
JP7132646B2 (ja) | 2022-09-07 |
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