Activating brown adipose tissue (BAT) could provide a potential approach for the treatment of obesity and metabolic disease in humans. Obesity is associated with upregulation of the endocannabinoid system, and blocking the cannabinoid type 1 receptor (CB1R) has been shown to cause weight loss and to decrease cardiometabolic risk factors. These effects may be mediated partly via increased BAT metabolism, since there is evidence that CB1R antagonism activates BAT in rodents. To investigate the significance of CB1R in BAT function, we quantified the density of CB1R in human and rodent BAT using the positron emission tomography radioligand [¹⁸F]FMPEP-d₂ and measured BAT activation in parallel with the glucose analog [¹⁸F]fluorodeoxyglucose. Activation by cold exposure markedly increased CB1R density and glucose uptake in the BAT of lean men. Similarly, β3-receptor agonism increased CB1R density in the BAT of rats. In contrast, overweight men with reduced BAT activity exhibited decreased CB1R in BAT, reflecting impaired endocannabinoid regulation. Image-guided biopsies confirmed CB1R mRNA expression in human BAT. Furthermore, CB1R blockade increased glucose uptake and lipolysis of brown adipocytes. Our results highlight that CB1Rs are significant for human BAT activity, and the CB1Rs provide a novel therapeutic target for BAT activation in humans.

Background: Copper-mediated radiofluorination is a straightforward method to produce a variety of [¹⁸F]fluoroarenes and [¹⁸F]fluoroheteroarenes. To minimize the number of steps in the production of ¹⁸F-labelled radiopharmaceuticals, we have developed a short and efficient azeotropic drying-free ¹⁸F-labelling method using copper-mediated fluorination. Our goal was to improve the copper-mediated method to achieve wide substrate scope with good radiochemical yields with short synthesis time. Results: Solid phase extraction with Cu (OTf)₂ in dimethylacetamide is a suitable activation method for [¹⁸F]fluoride. Elution efficiency with Cu (OTf)₂ is up to 79% and radiochemical yield (RCY) of a variety of model molecules in the crude reaction mixture has reached over 90%. Clinically relevant molecules, norepinephrine transporter tracer [¹⁸F]NS12137 and monoamine transporter tracer [¹⁸F]CFT were produced with 16.5% RCY in 98 min and 5.3% RCY in 64 min, respectively. Conclusions: Cu (OTf)₂ is a suitable elution agent for releasing [¹⁸F]fluoride from an anion exchange cartridge. The method is fast and efficient and the Cu-complex is customizable after the release of [¹⁸F]fluoride. Alterations in the [¹⁸F]fluoride elution techniques did not have a negative effect on the subsequent labelling reactions. We anticipate this improved [¹⁸F]fluoride elution technique to supplant the traditional azeotropic drying of [¹⁸F]fluoride in the long run and to concurrently enable the variations of the copper-complex.

Cannabinoid receptor 1 (CB1R) controls various physiological and pathological conditions, including memory, motivation, and inflammation and is thus an interesting target for positron emission tomography (PET). Herein, we report a ruthenium-mediated radiolabeling synthesis and preclinical evaluation of a new CB1R specific radiotracer, [¹⁸F]FPATPP. [¹⁸F]FPATPP was produced with 16.7 ± 5.7% decay-corrected radiochemical yield and >95 GBq/µmol molar activity. The tracer showed high stability, low defluorination and high specific binding to CB1Rs in mouse brain.

Aims/hypothesis: Stem cell-derived islets (SC-islets) are being used as cell replacement therapy for insulin-dependent diabetes. Non-invasive long-term monitoring methods for SC-islet grafts, which are needed to detect misguided differentiation in vivo and to optimise their therapeutic effectiveness, are lacking. Positron emission tomography (PET) has been used to monitor transplanted primary islets. We therefore aimed to apply PET as a non-invasive monitoring method for SC-islet grafts. Methods: We implanted different doses of human SC-islets, SC-islets derived using an older protocol or a state-of-the-art protocol and SC-islets genetically rendered hyper- or hypoactive into mouse calf muscle to yield different kinds of grafts. We followed the grafts with PET using two tracers, glucagon-like peptide 1 receptor-binding [¹⁸F]F-dibenzocyclooctyne-exendin-4 ([¹⁸F]exendin) and the dopamine precursor 6-[¹⁸F]fluoro-l-3,4-dihydroxyphenylalanine ([¹⁸F]FDOPA), for 5 months, followed by histological assessment of graft size and composition. Additionally, we implanted a kidney subcapsular cohort with different SC-islet doses to assess the connection between C-peptide and stem cell-derived beta cell (SC-beta cell) mass. Results: Small but pure and large but impure grafts were derived from SC-islets. PET imaging allowed detection of SC-islet grafts even <1 mm³ in size, [¹⁸F]exendin having a better detection rate than [¹⁸F]FDOPA (69% vs 44%, 1 mm³). Graft volume quantified with [¹⁸F]exendin (r²=0.91) and [¹⁸F]FDOPA (r²=0.86) strongly correlated with actual graft volume. [¹⁸F]exendin PET delineated large cystic structures and its uptake correlated with graft SC-beta cell proportion (r²=0.68). The performance of neither tracer was affected by SC-islet graft hyper- or hypoactivity. C-peptide measurements under fasted or glucose-stimulated conditions did not correlate with SC-islet graft volume or SC-beta cell mass, with C-peptide under hypoglycaemia having a weak correlation with SC-beta cell mass (r²=0.52). Conclusions/interpretation: [¹⁸F]exendin and [¹⁸F]FDOPA PET enable non-invasive assessment of SC-islet graft size and aspects of graft composition. These methods could be leveraged for optimising SC-islet cell replacement therapy in diabetes. Graphical

Here, we describe the development of an in‐house‐built device for the fully automated multistep synthesis of the cannabinoid CB₁ receptor imaging tracer (3R,5R)‐5‐(3‐([¹⁸F]fluoromethoxy‐d₂)phenyl)‐3‐(((R)‐1‐phenylethyl)amino)‐1‐(4‐(trifluoromethyl)phenyl)pyrrolidin‐2‐one ([¹⁸F]FMPEP‐d₂), following good manufacturing practices. The device is interfaced to a HPLC and a sterile filtration unit in a clean room hot cell. The synthesis involves the nucleophilic ¹⁸F‐fluorination of an alkylating agent and its GC purification, the subsequent ¹⁸F‐fluoroalkylation of a precursor molecule, the semipreparative HPLC purification of the ¹⁸F‐fluoroalkylated product, and its formulation for injection. We have optimized the duration and temperature of the ¹⁸F‐fluoroalkylation reaction and addressed the radiochemical stability of the formulated product. During the past 5 years (2013–2018), we have performed a total of 149 syntheses for clinical use with a 90% success rate. The activity yield of the formulated product has been 1.0 ± 0.4 GBq starting from 11 ± 2 GBq and the molar activity 600 ± 300 GBq/μmol at the end of synthesis.

[¹⁸F]NS12137 (exo‐3‐[(6‐[¹⁸F]fluoro‐2‐pyridyl)oxy]8‐azabicyclo[3.2.1]octane) is a highly selective norepinephrine transporter (NET) tracer. NETs are responsible for the reuptake of norepinephrine and dopamine and are linked to several neurodegenerative and neuropsychiatric disorders. The aim of this study was to develop a copper‐mediated ¹⁸F‐fluorination method for the production of [¹⁸F]NS12137 with straightforward synthesis conditions and high radiochemical yield and molar activity. [¹⁸F]NS12137 was produced in two steps. Radiofluorination of [¹⁸F]NS12137 was performed via a copper‐mediated pathway starting with a stannane precursor and using [¹⁸F]F⁻ as the source of the fluorine‐18 isotope. Deprotection was performed via acid hydrolysis. The radiofluorination reaction was nearly quantitative as was the deprotection based on HPLC analysis. The radiochemical yield of the synthesis was 15.1 ± 0.5%. Molar activity of [¹⁸F]NS12137 was up to 300 GBq/μmol. The synthesis procedure is straightforward and can easily be automated and adapted for clinical production.