Development and First-in-Human evaluation of a Site-Specific [F]-Labeled PD-L1 nanobody PET radiotracer for noninvasive imaging in NSCLC.

Immunohistochemistry (IHC) for PD-L1 detection is limited by its invasiveness and heterogeneity of tumors. To address these challenges, a new PD-L1-targeted nanobody-based immune-PET radiotracer [F]AlF-APN09 was developed using the site-specific radiolabeling method with the complexing agent (Mal-RESCA) under mild conditions. [F]AlF-APN09 was prepared at room temperature (pH 4.6-4.8) within 20 min with satisfactory radiochemical yields (45.8 ± 4.48 %, non-decay corrected), high radiochemical purity (>98 %) and moderate apparent molar activity (15-35 GBq/μmol), and remained stable in both PBS and 5 % HSA after 4 h (>90 %). Cell uptake studies indicated variable levels of surface PD-L1 expression in the following order: A549 > H1975 > A549. In micro-PET/CT imaging, A549 and H1975 tumors were distinctly visualized in a 6.0:1 and 3.2:1 ratios over PD-L1-negative A549 tumors in vivo. Ex vivo biodistribution studies showed tumor uptake values of 6.47 ± 1.06 %ID/g (A549) and 2.27 ± 0.19 %ID/g (H1975), significantly higher than 0.90 ± 0.28 %ID/g in A549 tumors. The estimated effective radiation dose in humans was 8.65E-03 mSv/MBq, lower than that of conventional [F]FDG. First-in-human imaging was conducted on a single resectable non-small cell lung cancer (NSCLC) subject without any adverse reactions. The radiotracer exhibited renal excretion with minimal hepatobiliary clearance. Tumor uptake reached SUV 4.20 at 2 h post-injection, demonstrating high contrast and rapid clearance. After PD-1 inhibitor immunotherapy and chemotherapy, the subject showed a therapeutic response and postoperative pathological specimens confirmed a major pathological response (MPR). These results suggest that we have successfully developed a new PD-L1-targeted nanobody PET tracer using the site-specific labeling method with the complexing agent (Mal-RESCA) within 20 min under mild conditions and [F]AlF-APN09 is a promising noninvasive PET radiotracer for visualizing PD-L1 expression in tumors, offering rapid tumor targeting, excellent signal-to-noise ratios, and favorable clearance properties.