Fast clearing RGD-based near-infrared fluorescent probes for in vivo tumor diagnosis

A fast clearing hydrophilic near-infrared (NIR) dye ICG-Der-02 was used to constitute tumor targeting contrast agents. Cell adhesion molecule integrin av beta 3 served as the target receptor because of its unique expression on almost all sprouting tumor vasculatures. The purpose of this study was to synthesize and compare the properties of integrin av beta 3-targeted, fast clearing NIR probes both in vitro and in vivo for tumor diagnosis. ICG-Der-02 was covalently conjugated to three kinds of RGD peptide including linear, monoeric cyclic and dimeric RGD to form three RGD-based NIR probes. The integrin receptor specificities of these probes were evaluated in vitro by confocal microscopy. The dynamic bio-distribution and elimination ratse were in vivo real-time monitored by a near-infrared imaging system in normal mice. Further, the in vivo tumor targeting abilities of the RGD-based NIR probes were compared in av beta 3-positive MDA-MB-231, U87MG and alpha(v)beta 3-negtive MCF-7 xenograft mice models. Three RGD-based NIR probes were successfully synthesized with good optical properties. In vitro cellular experiments indicated that the probes have a clear binding affinity to alpha(U)beta 3-positive tumor cells, with a cyclic dimeric RGD probe owing the highest integrin affinity. Dynamic bio-distributions of these probes showed a rapid clearing rate through the renal pathway. In vivo tumor targeting ability of the RGD-based porbes was demonstrated on MDA-MB-231 and U87MG tumor models. As expected, the c(RGDyK)2-ICG-Der-02 probe displayed the highest tumor-to-normal tissue contrast. The in vitro and in vivo block experiments confirmed the receptor binding specificity of the probes. The hydrophilic dye-labeled NIR probes exhibited a fast clearing rate and deep tissue penetration capability. Further, the alpha(U)beta 3 receptor affinity of the three RGD-based NIR probes followed the order of dimer cyclic > monomer cyclic > linear. The results demonstrate potent fast clearing probes for in vivo early tumor diagnosis. Copyright (c) 2012 John Wiley & Sons, Ltd.