Abstract
BackgroundThe P2X7 receptor has been shown to play a fundamental role in the initiation and sustenance of the inflammatory cascade. The development of a novel fluorine-18 PET tracer superior and with a longer half-life to those currently available is a promising step towards harnessing the therapeutic and diagnostic potential offered by this target. Inspired by the known antagonist A-804598, the present study outlines the design via molecular docking, synthesis and biological evaluation of the novel P2X7 tracer [18F]EFB. The tracer was radiolabelled via a three-step procedure, in vitro binding assessed in P2X7-transfected HEK293 and in B16 cells by calcium influx assays and an initial preclinical evaluation was performed in a lipopolysaccharide (LPS)-injected rat model of neuroinflammation.ResultsThe novel tracer [18F]EFB was synthesised in 210 min in 3–5% decay-corrected radiochemical yield (DC RCY), >99% radiochemical purity (RCP) and >300 GBq/μmol and fully characterised. Functional assays showed that the compound binds with nM Ki to human, rat and mouse P2X7 receptors. In vivo, [18F]EFB displayed a desirable distribution profile, and while it showed low blood–brain barrier penetration, brain uptake was quantifiable and displayed significantly higher mean longitudinal uptake in inflamed versus control rat CNS regions.Conclusions[18F]EFB demonstrates strong in vitro affinity to human and rodent P2X7 and limited yet quantifiable BBB penetration. Considering the initial promising in vivo data in an LPS rat model with elevated P2X7 expression, this work constitutes an important step in the development of a radiotracer useful for the diagnosis and monitoring of clinical disorders with associated neuroinflammatory processes.
Highlights
The P2X7 receptor has been shown to play a fundamental role in the initiation and sustenance of the inflammatory cascade
Neither [11C]A740003 nor [11C]SMW64-D16 showed appreciable brain uptake; both succeeded in recognising the site of inflammation in two different rodent models when examined by in vitro autoradiography [5, 6]. [11C]GSK1482160, shows greater promise due to its efficient radiosynthesis and strong P2X7 selectivity and blood–brain barrier (BBB) permeability, but its preclinical evaluation is still in progress [7]
This study presents the novel fluorine-18-labelled PET tracer [18F]EFB 2-cyano-1-(4-[18F]fluorobenzyl)-3-(quinolin-5-yl)guanidine developed from the known blood–brain barrier permeable P2X7 antagonist A-804598 [8, 9]
Summary
The P2X7 receptor has been shown to play a fundamental role in the initiation and sustenance of the inflammatory cascade. The tracer was radiolabelled via a three-step procedure, in vitro binding assessed in P2X7-transfected HEK293 and in B16 cells by calcium influx assays and an initial preclinical evaluation was performed in a lipopolysaccharide (LPS)-injected rat model of neuroinflammation. In vivo, [18F]EFB displayed a desirable distribution profile, and while it showed low blood–brain barrier penetration, brain uptake was quantifiable and displayed significantly higher mean longitudinal uptake in inflamed versus control rat CNS regions. Considering the initial promising in vivo data in an LPS rat model with elevated P2X7 expression, this work constitutes an important step in the development of a radiotracer useful for the diagnosis and monitoring of clinical disorders with associated neuroinflammatory processes. [11C]GSK1482160, shows greater promise due to its efficient radiosynthesis and strong P2X7 selectivity and blood–brain barrier (BBB) permeability, but its preclinical evaluation is still in progress [7]. Neither [11C]A740003 nor [11C]SMW64-D16 showed appreciable brain uptake; both succeeded in recognising the site of inflammation in two different rodent models when examined by in vitro autoradiography [5, 6]. [11C]GSK1482160, shows greater promise due to its efficient radiosynthesis and strong P2X7 selectivity and blood–brain barrier (BBB) permeability, but its preclinical evaluation is still in progress [7].
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