An RGD modified water-soluble fluorophore probe for in vivo NIR-II imaging of thrombosis.

Venous thrombosis leads to severe symptoms and death through pulmonary embolism. There is a great need for high sensitivity imaging methods to identify acute patients who would benefit from thrombolysis. We designed a novel, organic near-infrared second-window (NIR-II) probe, which targets the glycoprotein IIb/IIIa receptor (GPIIb/IIIa) on activated platelets. The probe's structure was characterized by MALDI-TOF-MS, TEM, UV-visible absorption and NIR-II fluorescent spectroscopy. The probe's specificity for activated platelets was investigated in vitro and in vivo. Thrombosis in mice was induced by administration of FeCl in the external jugular vein and imaged by using a NIR-II imager. The donor-acceptor-donor fluorescent core TTQ was prepared from donor and acceptor units. TTQ-PEG-NH was synthesized by sequential modification of PEGylated TTQ, followed by c(RGD) condensation. Signal strength was continuously monitored for 24 h following TTQ-PEG-c(RGD) or non-specific fluorescent dye injection. The contralateral external jugular vein, sham surgery and a competitive inhibition experiment served as controls. TTQ-PEG-c(RGD) presented high NIR-II intensity, good stability and excellent affinity for activated platelets. The NIR-II fluorescence signal of TTQ-PEG-c(RGD) injected mice significantly increased at the thrombus site and peaked at 4 h, whereas there was no significant change in the control mice, and the competitive inhibition of the RGD antagonist suppressed the enhancement of the NIR-II fluorescence signal. Comparison between fresh and old thrombi confirmed that TTQ-PEG-c(RGD) could be used to distinguish a fresh thrombus from an old thrombus. TTQ-PEG-c(RGD) can specifically target thrombosis in vitro and in vivo, providing a potential tool for noninvasive diagnosis of early thrombi.