Development of a Novel Gene Silencer Pyrrole-Imidazole Polyamide Targeting Human Connective Tissue Growth Factor

• Wan Jian-Xin

  Division of Nephrology Hypertension and Endocrinology, Department of Medicine, School of Medicine, Nihon University

• Fukuda Noboru

  Division of Nephrology Hypertension and Endocrinology, Department of Medicine, School of Medicine, Nihon University Advanced Research Institute of the Sciences and Humanities, Nihon University

• Ueno Takahiro

  Division of Nephrology Hypertension and Endocrinology, Department of Medicine, School of Medicine, Nihon University

• Watanabe Takayoshi

  Division of Cancer Genetics, Department of Advanced Medical Science, School of Medicine, Nihon University

• Matsuda Hiroyuki

  Advanced Research Institute of the Sciences and Humanities, Nihon University

• Saito Kosuke

  Advanced Research Institute of the Sciences and Humanities, Nihon University

• Nagase Hiroki

  Advanced Research Institute of the Sciences and Humanities, Nihon University Division of Cancer Genetics, Department of Advanced Medical Science, School of Medicine, Nihon University

• Matsumoto Yoshiaki

  Department of Clinical Pharmacokinetics, School of Pharmacy, Nihon University

• Matsumoto Koichi

  Division of Nephrology Hypertension and Endocrinology, Department of Medicine, School of Medicine, Nihon University

抄録

Pyrrole–imidazole (PI) polyamide can bind to specific sequences in the minor groove of double-helical DNA and inhibit transcription of the genes. We designed and synthesized a PI polyamide to target the human connective tissue growth factor (hCTGF) promoter region adjacent to the Smads binding site. Among coupling activators that yield PI polyamides, 1-[bis(dimethylamino)methylene]-5-chloro-1H-benzotriazolium 3-oxide hexafluorophosphate (HCTU) was most effective in total yields of PI polyamides. A gel shift assay showed that a PI polyamide designed specifically for hCTGF (PI polyamide to hCTGF) bound the appropriate double-stranded oligonucleotide. A fluorescein isothiocyanate (FITC)-conjugated PI polyamide to CTGF permeated cell membranes and accumulated in the nuclei of cultured human mesangial cells (HMCs) and remained there for 48 h. The PI polyamide to hCTGF significantly decreased phorbol 12-myristate acetate (PMA)- or transforming growth factor-β1 (TGF-β1)-stimulated luciferase activity of the hCTGF promoter in cultured HMCs. The PI polyamide to hCTGF significantly decreased PMA- or TGF-β1-stimulated expression of hCTGF mRNA in a dose-dependent manner. The PI polyamide to hCTGF significantly decreased PMA- or TGF-β1-stimulated levels of hCTGF protein in HMCs. These results indicate that the developed synthetic PI polyamide to hCTGF could be a novel gene silencer for fibrotic diseases.

収録刊行物

• Ｂｉｏｌｏｇｉｃａｌ　＆　Ｐｈａｒｍａｃｅｕｔｉｃａｌ　Ｂｕｌｌｅｔｉｎ

  Ｂｉｏｌｏｇｉｃａｌ　＆　Ｐｈａｒｍａｃｅｕｔｉｃａｌ　Ｂｕｌｌｅｔｉｎ 34 (10), 1572-1577, 2011

  公益社団法人 日本薬学会

キーワード

• pyrrole–imidazole polyamide
• connective tissue growth factor
• promoter activity
• Fmoc method
• gene therapy
• human

詳細情報

• CRID: 1390001204631335552
• NII論文ID: 130001086589
• NII書誌ID: AA10885497
• DOI: 10.1248/bpb.34.1572
• ISSN: 13475215
• ISSN: 09186158
• NDL書誌ID: 11250101
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I11250101
• Web Site: http://www.jstage.jst.go.jp/article/bpb/34/10/34_10_1572/_pdf
• Web Site: https://search.jamas.or.jp/link/ui/2012139288
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • CiNii Articles
  • KAKEN
• 抄録ライセンスフラグ: 使用不可