Inorganic Chemistry I, Bioinorganic Chemistry, Faculty of Chemistry and Biochemistry, Ruhr-University Bochum, Universitätsstrasse 150, D-44801 Bochum, Germany.
Bioconjug Chem. 2012 Sep 19;23(9):1764-74. doi: 10.1021/bc200692g. Epub 2012 Aug 30.
Labeling of peptide nucleic acids (PNA) with metallocene complexes is explored herein for the modulation of the analytical characteristics, as well as biological properties of PNA. The synthesis of the first ruthenocene-PNA conjugate with a dodecamer, mixed-sequence PNA is described, and its properties are compared to a ferrocene-labeled analogue as well as an acetylated, metal-free derivative. The synthetic characteristics, chemical stability, analytical and thermodynamic properties, and the interaction with cDNA were investigated. Furthermore, the cytotoxicity of the PNA conjugates is determined on HeLa, HepG2, and PT45 cell lines. Finally, the cellular uptake of the metal-containing PNAs was quantified by high-resolution continuum source atomic absorption spectrometry (HR-CS AAS). An unexpectedly high cellular uptake to final concentrations of 4.2 mM was observed upon incubation with 50 μM solutions of the ruthenocene-PNA conjugate. The ruthenocene label was shown to be an excellent label in all respects, which is also more stable than its ferrocene analogue. Because of its high stability, low toxicity, and the lack of a natural background of ruthenium, it is an ideal choice for bioanalytical purposes and possible medicinal and biological applications like, e.g., the development of gene-targeted drugs.
本文探索了将金属茂配合物标记肽核酸（PNA），以调节 PNA 的分析特性和生物学特性。描述了第一个带有十二聚体、混合序列 PNA 的钌茂-PNA 缀合物的合成，并将其性质与铁茂标记类似物以及乙酰化、无金属衍生物进行了比较。研究了合成特性、化学稳定性、分析和热力学性质以及与 cDNA 的相互作用。此外，还测定了 PNA 缀合物在 HeLa、HepG2 和 PT45 细胞系上的细胞毒性。最后，通过高分辨率连续光源原子吸收光谱法（HR-CS AAS）定量测定了含金属 PNA 的细胞摄取量。在用 50 μM 溶液孵育时，观察到出人意料的高细胞摄取终浓度为 4.2 mM。钌茂标记在各方面都是一个极好的标记，而且比其铁茂类似物更稳定。由于其高稳定性、低毒性以及缺乏天然的钌背景，它是生物分析目的以及可能的医学和生物学应用的理想选择，例如，开发基因靶向药物。
