Synthesis and Evaluation of ^99mTc-Labeled FAP Inhibitors with Different Linkers for Imaging of Fibroblast Activation Proteins in Tumors

Abstract

Fibroblast activation protein (FAP) is a potential target for tumor diagnosis and treatment due to its selective expression on cancer-associated fibroblasts (CAFs) in most solid tumor stroma. Two FAP inhibitor (FAPI) derived ligands (L1 and L2) containing different lengths of _DPro-Gly (PG) repeat units as linkers were designed and synthesized with high affinity for FAP. Two stable hydrophilic ^99mTc-labeled complexes ([^99mTc]Tc-L1 and [^99mTc]Tc-L2) were obtained. <i>In vitro</i> cellular studies show that the uptake mechanism is correlated with FAP uptake, and [^99mTc]Tc-L1 shows a higher cell uptake and specific binding to FAP. A nanomolar <i>K</i>_d value for [^99mTc]Tc-L1 indicates its significantly high target affinity for FAP. The biodistribution and microSPECT/CT images obtained for U87MG tumor mice show that [^99mTc]Tc-L1 has high tumor uptake with specificity to FAP and high tumor-to-nontarget ratios. As an inexpensive, easily made, and widely available tracer, [^99mTc]Tc-L1 holds great promise for clinical applications.