Synthesis and Preliminary Study of ^99mTc-Labeled HYNIC-FAPi for Imaging of Fibroblast Activation Proteins in Tumors

Abstract

Fibroblast activation protein (FAP) is an emerging target for cancer diagnosis. Different types of FAP inhibitor (FAPI)-based radiotracers have been developed and applied for tumor imaging. However, few FAPI tracers for single photon emission computed tomography (SPECT) imaging have been reported. SPECT imaging is less expensive and more widely distributed than positron emission tomography (PET), and thus, 99m Tclabeled FAPIs would be more available to patients in developing regions. Herein, we developed a FAPI-04-derived radiotracer, HYNIC-FAPi-04 (HFAPi), for SPECT imaging. 99m Tc-HFAPi, with a radiochemical purity of >98%, was prepared using a kit formula within 30 min. The specificity of 99m Tc-HFAPi for FAP was validated by a cell binding assay in vitro and SPECT/CT imaging in vivo. The binding affinity (K d value) of 99m Tc-HFAPi for human FAP and murine FAP was 4.49 and 2.07 nmol/L, respectively. SPECT/CT imaging in HT1080-hFAP tumor-bearing mice showed the specific FAP targeting ability of 99m Tc-HFAPi in vivo. In U87MG tumor-bearing mice, 99m Tc-HFAPi had a higher tumor uptake compared with that of HT1080-hFAP and 4T1-mFAP tumor models. Interestingly, 99m Tc-HFAPi showed a relatively high uptake in some murine joints. 99m Tc-HFAPi accumulated in tumor lesions with a high tumor-to-background ratio. A preliminary clinical study was also performed in breast cancer patients. Additionally, 99m Tc-HFAPi exhibited an advantage over 18 F-FDG in the detection of lymph node metastatic lesions in breast cancer patients, which is helpful in improving treatment strategies. In short, 99m Tc-HFAPi showed excellent affinity and specificity for FAP and is a promising SPECT radiotracer for (re)staging and treatment planning of breast cancers.

Radiolabelled FGF-2 for Imaging Activated Fibroblasts in the Tumor Micro-Environment

“…For these reasons, in recent years, small molecules that inhibit this protein have gained interest in detecting and treating tumors. Indeed, FAP-specific enzyme inhibitors (FAPIs) have demonstrated high specificity and affinity and good pharmacokinetic profiles in both preclinical and clinical phases [31][32][33][34][35]. Indeed, in the last 10 years, several new FAPI-based radiopharmaceuticals have been developed, with promising results for diagnostic and therapeutic purposes [36][37][38][39][40][41][42].…”