Access to 18F-Labeled FAP Inhibitor variants via [18F]SuFEx Reaction

´Objectives:

The emerging significance of the tumor microenvironment (TME) as a new frontier for cancer diagnosis and therapy can be primarily attributed to its unique features, such as the interconnection between stromal and cancer cells.1 Cancer-associated fibroblasts (CAFs) within the TME are identified by biomarkers such as fibroblast activation protein alpha (FAP), which are expressed on their surfaces. Targeting FAP using small molecule 18F-labeled inhibitors (FAPIs) have recently garnered significant attention for noninvasive tumor visualization using PET.2 Currently, the predominant 18F-fluorination method for radiolabeling FAPIs involves chelation-based radiofluorination strategies using aluminum [18F]fluoride ([18F]AlF). Herein, a powerful radiofluorination protocol for the preparation of an 18F-labeled FAPI via the sulfur [18F]fluoride exchange ([18F]SuFEx) reaction is disclosed.3
Methods:

The incorporation of the aryl fluorosulfate motif into the linker of the FAPI core structure (2) via amide bond formation allowed the radiolabeling precursor 3 to be accessed in moderate yield (Scheme 1 A, 46%). The radiosynthesis commenced with [18F]fluoride loading onto a QMA-cartridge which was eluted with a methanolic solution containing Et4NHCO3, followed by evaporation of the solvent under reduced pressure at 70 oC for 5 min (Scheme 1 B). Thereafter, the precursor 3 (100 µg, 145 nmol) in MeCN was added to the reaction vial, and allowed to react by [18F]SuFEx at room temperature for 5 min. The reaction was quenched by water dilution followed by SPE-based purification using a C18 cartridge. [18F]3 was isolated by elution from the cartridge with EtOH and the identity of the product was confirmed by UHPLC.

Results

The optimized radiosynthesis of 18F-labeled FAPI ([18F]3) was obtained with non-decay corrected isolated activity yields (AY) of 54 ± 3% (n = 3) and >99% RCP in 25 min. The automated radiosynthesis afforded [18F]3 in an unoptimized 11% AY, with >95% RCP and molar activity (Am) of 25 GBq/µmol (n = 1) in 30 min. The product was obtained in 2 mL EtOH, which can easily be further diluted with water or saline solution for subsequent biological evaluation.