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The evaluation of new and isotopically labeled isoindoline nitroxides and an azaphenalene nitroxide for EPR oximetry

机译:评估新的和同位素标记的异吲哚啉氮氧化物和氮杂氮烯氮氧化物用于EPR血氧饱和度测定

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Isoindoline nitroxides are potentially useful probes for viable biological systems, exhibiting low cytotoxicity, moderate rates of biological reduction and favorable Electron Paramagnetic Resonance (EPR) characteristics. We have evaluated the anionic (5-carboxy-1,1,3,3-tetramethylisoindolin-2-yloxyl; CTMIO), cationic (5-(N,N,N-trimethylammonio)-1,1,3,3-tetramethylisoindolin-2-yloxyl iodide, QATMIO) and neutral (1,1,3,3-tetramethylisoindolin-2-yloxyl; TMIO) nitroxides and their isotopically labeled analogs (~2H_(12)- and/or ~2H_(12)-~15N-labeled) as potential EPR oximetry probes. An active ester analogue of CTMIO, designed to localize intracellularly, and the azaphenalene nitroxide 1,1,3,3-tetramethyl-2,3-dihydro- 2-azaphenalen-2-yloxyl (TMAO) were also studied. While the EPR spectra of the unlabeled nitroxides exhibit high sensitivity to O_2 concentration, deuteration resulted in a loss of superhyperfine features and a subsequent reduction in O_2 sensitivity. Labeling the nitroxides with ~(15)N increased the signal intensity and this may be useful in decreasing the detection limits for in vivo measurements. The active ester nitroxide showed approximately 6% intracellular localization and low cytotoxicity. The EPR spectra of TMAO nitroxide indicated an increased rigidity in the nitroxide ring, due to dibenzo-annulation.
机译:异吲哚啉氮氧化物是可行的生物系统潜在探针,具有低细胞毒性,中等程度的生物还原速率和良好的电子顺磁共振(EPR)特性。我们已经评估了阴离子(5-羧基-1,1,3,3-四甲基异吲哚-2-基氧基; CTMIO),阳离子(5-(N,N,N-三甲基铵)-1,1,3,3-四甲基异吲哚啉-2-基碘碘化物(QATMIO)和中性(1,1,3,3-四甲基异吲哚-2-基氧基; TMIO)氮氧化物及其同位素标记的类似物(〜2H_(12)-和/或〜2H_(12)-〜 15N标记)作为潜在的EPR血氧饱和度探针。还研究了设计用于细胞内定位的CTMIO的活性酯类似物,以及氮杂氮烯的氮氧化物1,1,3,3-四甲基-2,3-二氢-2-氮杂苯并-2-基氧基(TMAO)。尽管未标记的氮氧化物的EPR光谱对O_2浓度表现出高敏感性,但是氘化导致超高精细特征的丧失和随后O_2敏感性的降低。用〜(15)N标记一氧化氮可增加信号强度,这可能有助于降低体内测量的检测限。活性酯氮氧化物显示约6%的细胞内定位和低细胞毒性。 TMAO氮氧化物的EPR光谱表明,由于二苯并环化,氮氧化物环的刚性增加。

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