Synthesis and characterization of a Cu(ii) coordination-containing TAM radical as a nitroxyl probe

Nitroxyl (HNO) has been identified as an important signaling molecule in biological systems, and it plays critical roles in many physiological processes. However, its detection remains challenging because of the limited sensitivity and/or specificity of existing detection methods. Low-frequency electron paramagnetic resonance (EPR) spectroscopy and imaging, coupled with the use of exogenous paramagnetic probes, have been indispensable techniques for the in vivo measurement of various physiological parameters owing to their specificity, noninvasiveness and good depth of magnetic field penetration in animal tissues. However, the in vivo detection of HNO levels by EPR spectroscopy and imaging is limited due to the need for improved probes. We report the first "turn on-response" EPR probe for HNO utilizing a Cu(ii) coordination-containing TAM radical (denoted as Cu^II[TD1]). Upon reaction with HNO, Cu^II[TD1] shows a 16.1-fold turn-on in EPR signal with a low detection limit of 1.95 μM. Moreover, low-temperature EPR spectroscopic and ESI-MS studies showed that the sensing mechanism relies on the reduction of Cu(ii) by HNO. Lastly, Cu^II[TD1] is selective for HNO over other reactive nitrogen and oxygen species except for some reductants (Cys and Asc). This new Cu(ii) coordination-containing TAM radical shows great potential for in vivo EPR HNO applications in the absence of reducing agents and provides insights into developing improved and targeted EPR HNO probes for biomedical applications.