[show abstract][hide abstract] ABSTRACT: The PET radioligand (11)C-CUMI-101 was previously suggested as a putative agonist radioligand for the serotonin 1A (5-hydroxytryptamine 1A [5-HT1A]) receptor in recombinant cells expressing human 5-HT1A receptor. However, a recent study showed that CUMI-101 behaved as a potent 5-HT1A receptor antagonist in rat brain. CUMI-101 also has moderate affinity (Ki = 6.75 nM) for α1 adrenoceptors measured in vitro. The current study examined the functional properties and selectivity of CUMI-101, both in vitro and in vivo.
The functional assay was performed using (35)S-GTPγS (GTP is guanosine triphosphate) in primate brains. The cross-reactivity of CUMI-101 with α1 adrenoceptors was performed using in vitro radioligand binding studies in rat, monkey, and human brains as well as in vivo PET imaging in mouse, rat, and monkey brains.
CUMI-101 did not stimulate (35)S-GTPγS binding in primate brain, in contrast to 8-OH-DPAT, a potent 5-HT1A receptor agonist. Instead, CUMI-101 behaved as a potent 5-HT1A receptor antagonist by dose-dependently inhibiting 8-OH-DPAT-stimulated (35)S-GTPγS binding. Both in vitro and in vivo studies showed that CUMI-101 had significant α1 adrenoceptor cross-reactivity. On average, across all 3 species examined, cross-reactivity was highest in the thalamus (>45%) and lowest in the neocortex and cerebellum (<10%). PET imaging further confirmed that only preblocking with WAY-100635 plus prazosin decreased (11)C-CUMI-101 brain uptake to that of self-block.
CUMI-101 behaves as a 5-HT1A receptor antagonist in primate brain, with significant, regional-dependent α1 adrenoceptor cross-reactivity, limiting its potential use as a PET radioligand in humans.
Journal of Nuclear Medicine 01/2014; 55(1):141-6. · 5.77 Impact Factor
[show abstract][hide abstract] ABSTRACT: Positron emission tomography (PET) has growing importance as a molecular imaging technique in clinical research and drug development. Methods for producing PET radiotracers utilizing cyclotron-produced [(18)F]fluoride ion (t1/2 = 109.7 min) without the need for complete removal of irradiated target [(18)O]water and addition of cryptand are keenly sought for practical convenience and efficiency. Several structurally diverse diaryliodonium tosylates, XArI(+)Ar'Y TsO(-) (X = H or p-MeO), were investigated in a microfluidic apparatus for their reactivity towards radiofluorination with high specific activity (no-carrier-added) [(18)F]fluoride ion in mixtures of DMF and irradiated target [(18)O]water in the absence of cryptand. Salts bearing a para or ortho electron-withdrawing group Y (e.g., Y = p-CN) reacted rapidly (∼3 min) to give the expected major [(18)F]fluoroarene product, [(18)F]FArY, in useful moderate radiochemical yields even when the solvent had an [(18)O]water content up to 28%. Salts bearing electron-withdrawing groups in meta position (e.g., Y = m-NO2), or an electron-donating substituent (Y = p-OMe), gave low radiochemical yields under the same conditions.
[show abstract][hide abstract] ABSTRACT: Here we aimed to explore the feasibility of enhancing the fluorination of organosilanes by appending potassium-chelating groups to the substrates. For this purpose, eight organosilanes were prepared in which a linear or cyclic leaving group, with putative potassium-chelating ability, was attached covalently to a congested silicon atom via an ether linkage to serve as a potential nucleophilic assisting leaving group (NALG). Organosilicon-NALGs with expected strong potassium-chelating capability enhanced reactions with potassium fluoride in acetonitrile to produce organfluorosilanes without any need to separately add phase transfer reagent. Similar rate enhancements were also observed with cyclotron-produced [18F]fluoride ion (t1/2 = 109.7 min, β+ = 97%) in the presence of potassium carbonate in MeCN-0.5% H2O. This study found that metal-chelating NALG units can accelerate fluorination and radiofluorination reactions at sterically crowded silicon atoms.
Journal of Fluorine Chemistry 01/2013; · 1.94 Impact Factor
[show abstract][hide abstract] ABSTRACT: Rhodamine-123 is a known substrate for the efflux transporter, P-glycoprotein (P-gp). We wished to assess whether rhodamine-123 might serve as a useful substrate for developing probes for imaging efflux transporters in vivo with positron emission tomography (PET). For this purpose, we aimed to label rhodamine-123 with carbon-11 (t(1/2)=20.4min) and to study its biodistribution in rodents.
[(11)C]Rhodamine-123 was prepared by treating rhodamine-110 (desmethyl-rhodamine-123) with [(11)C]methyl iodide. The biodistribution of this radiotracer was studied with PET in wild-type mice and rats, in efflux transporter knockout mice, in wild-type rats pretreated with DCPQ (an inhibitor of P-gp) or with cimetidine (an inhibitor of organic cation transporters; OCT), and in P-gp knockout mice pretreated with cimetidine. Unchanged radiotracer in forebrain, plasma and peripheral tissues was also measured ex vivo at 30min after radiotracer administration to wild-type and efflux transporter knockout rodents.
[(11)C]Rhodamine-123 was obtained in 4.4% decay-corrected radiochemical yield from cyclotron-produced [(11)C]carbon dioxide. After intravenous administration of [(11)C]rhodamine-123 to wild-type rodents, PET and ex vivo measurements showed radioactivity uptake was very low in brain, but relatively high in some other organs such as heart, and especially liver and kidney. Inhibition of P-gp increased uptake in brain, heart, kidney and liver, but only by up to twofold. Secretion of radioactivity from kidney was markedly reduced by OCT knockout or pretreatment with cimetidine.
[(11)C]Rhodamine-123 was unpromising as a PET probe for P-gp function and appears to be a strong substrate of OCT in kidney. Cimetidine appears effective for blocking OCT in kidney in vivo.
Nuclear Medicine and Biology 08/2012; 39(8):1128-36. · 2.52 Impact Factor
[show abstract][hide abstract] ABSTRACT: A known chemotype of H(3) receptor ligand was explored for development of a radioligand for imaging brain histamine subtype 3 (H(3)) receptors in vivo with positron emission tomography (PET), namely nonimidazole 2-aminoethylbenzofurans, represented by the compound (R)-(2-(2-(2-methylpyrrolidin-1-yl)ethyl)benzofuran-5-yl)(4-fluorophenyl)methanone (9). Compound 9 was labeled with fluorine-18 (t(1/2) = 109.7 min) in high specific activity by treating the prepared nitro analogue (12) with cyclotron-produced [(18)F]fluoride ion. [(18)F]9 was studied with PET in mouse and in monkey after intravenous injection. [(18)F]9 showed favorable properties as a candidate PET radioligand, including moderately high brain uptake with a high proportion of H(3) receptor-specific signal in the absence of radiodefluorination. The nitro compound 12 was found to have even higher H(3) receptor affinity, indicating the potential of this chemotype for the development of further promising PET radioligands.
Journal of Medicinal Chemistry 03/2012; 55(5):2406-15. · 5.61 Impact Factor
[show abstract][hide abstract] ABSTRACT: Rapid and direct: the carboxylation of boronic acid esters with (11)CO(2) provides [(11)C]carboxylic acids as a convenient entry into [(11)C]esters and [(11)C]amides. This conversion of boronates is tolerant to diverse functional groups (e.g., halo, nitro, or carbonyl).
Angewandte Chemie International Edition 02/2012; 51(11):2698-702. · 13.73 Impact Factor
[show abstract][hide abstract] ABSTRACT: 3-Fluoro-1-((thiazol-4-yl)ethynyl)benzenes constitute an important class of high-affinity metabotropic glutamate subtype 5 receptor (mGluR5) ligands, some of which have been labeled with fluorine-18 (t(1/2) = 109.7 min), to provide radioligands for molecular imaging of brain mGluR5 in living animal and human subjects with positron emission tomography (PET). Labeling in the 3-fluoro position of such ligands can be achieved through aromatic nucleophilic substitution of a halide leaving group with [(18)F]fluoride ion when a weakly activating m-nitrile group is present, but is generally very low yielding (<8%). Here we used a microfluidic reaction platform to show that greatly enhanced (up to 6-fold) radiochemical yields can be achieved from suitably synthesized diaryliodonium tosylate precursors. The presence of a m-nitrile or other activating group is not required. Similar conditions were adopted in a more conventional automated radiochemistry platform having a single-pot reactor, to produce mGluR5 radioligands with useful radioactivities for PET imaging.
[show abstract][hide abstract] ABSTRACT: Effective methods for the introduction of the short-lived positron-emitter fluorine-18 (t(1/2) = 109.7 min) at high specific radioactivity into fluoroarenes are valuable for the development of radiotracers for molecular imaging with positron emission tomography. Here we have explored the scope of the radiofluorination of diaryliodonium salts with no-carrier-added [(18)F]fluoride ion for the preparation of otherwise difficult to access meta-substituted [(18)F]fluoroarenes. A microfluidic reaction platform was used to establish optimal radiochemical yields. Rapid, high yielding and selective radiofluorinations were achieved in unsymmetrical diaryliodonium tosylates (ArI(+)Ar'TsO(-)) in which Ar carried either a meta electron-withdrawing (CN, NO(2), CF(3)) or a meta electron-donating (Me or MeO) group, and in which the partner aryl group (Ar') was relatively electron-rich, such as Ph, 3-Me-C(6)H(4), 4-MeO-C(6)H(4), 2-thienyl or 5-Me-2-thienyl. The radiofluorination of appropriate diaryliodonium tosylates is therefore a generally useful method for the preparation of simple [(18)F]m-fluoroarenes ([(18)F]ArF).
Annalen der Chemie und Pharmacie 06/2011; 2011(23):4439 - 4447. · 3.10 Impact Factor
[show abstract][hide abstract] ABSTRACT: [(18)F]Xenon difluoride ([(18)F]XeF(2)), was produced by treating xenon difluoride with cyclotron-produced [(18)F]fluoride ion to provide a potentially useful agent for labeling novel radiotracers with fluorine-18 (t(1/2) = 109.7 min) for imaging applications with positron emission tomography, Firstly, the effects of various reaction parameters, for example, vessel material, solvent, cation and base on this process were studied at room temperature. Glass vials facilitated the reaction more readily than polypropylene vials. The reaction was less efficient in acetonitrile than in dichloromethane. Cs(+) or K(+) with or without the cryptand, K 2.2.2, was acceptable as counter cation. The production of [(18)F]XeF(2) was retarded by K(2)CO(3), suggesting that generation of hydrogen fluoride in the reaction milieu promoted the incorporation of fluorine-18 into xenon difluoride. Secondly, the effect of temperature was studied using a microfluidic platform in which [(18)F]XeF(2) was produced in acetonitrile at elevated temperature (≥ 85 °C) over 94 s. These results enabled us to develop a method for obtaining [(18)F]XeF(2) on a production scale (up to 25 mCi) through reaction of [(18)F]fluoride ion with xenon difluoride in acetonitrile at 90 °C for 10 min. [(18)F]XeF(2) was separated from the reaction mixture by distillation at 110 °C. Furthermore, [(18)F]XeF(2) was shown to be reactive towards substrates, such as 1-((trimethylsilyl)oxy)cyclohexene and fluorene.
Journal of Fluorine Chemistry 10/2010; 131(10):1032-1038. · 1.94 Impact Factor
[show abstract][hide abstract] ABSTRACT: [carbonyl-(11)C]Benzyl acetate ([(11)C]1) has been proposed as a potential agent for imaging glial metabolism of acetate to glutamate and glutamine with positron emission tomography (PET). [(11)C]1 was synthesized from [(11)C]carbon monoxide, iodomethane and benzyl alcohol via palladium-mediated chemistry. The radiosynthesis was automated with a modified Synthia platform controlled with in-house developed Labview software. Under production conditions, [(11)C]1 was obtained in 10% (n = 6) decay-corrected radiochemical yield from [(11)C]carbon monoxide in > 96% radiochemical purity and with an average specific radioactivity of 2,415 mCi/micromol. The total radiosynthesis time was about 45 min. Peak uptake of radioactivity in brain (SUV = 3.1) was relatively high and may be amenable to measuring uptake and metabolism of acetate in glial cells of the brain.
Journal of Labelled Compounds 07/2010; 53(8):548 - 551.
[show abstract][hide abstract] ABSTRACT: Dopamine released by amphetamine decreases the in vivo binding of PET radioligands to the dopamine D(2) receptor. Although concentrations of extracellular dopamine largely return to baseline within 1 to 2 h after amphetamine treatment, radioligand binding remains decreased for several hours. The purpose of this study was to determine whether the prolonged decrease of radioligand binding after amphetamine administration is caused by receptor internalization. To distinguish dopamine displacement from receptor internalization, we used wild-type and arrestin3 (arr3) knockout mice, which are incapable of internalizing D(2) receptors. In addition, we used both the D(2) selective agonist [(11)C]MNPA (which is thought to bind to the high affinity state of the receptor) and the D(2) selective antagonist [(18)F]fallypride (which does not differentiate between high and low affinity state). After an initial baseline scan, animals were divided in three groups for a second scan: either 30 min or 4 h after amphetamine administration (3 mg/kg, i.p.) or as retest. At 30 min, [(11)C]MNPA showed greater displacement than [(18)F]fallypride, but each radioligand gave similar displacement in knockout and wild-type mice. At 4 h, the binding of both radioligands returned to baseline in arr3 knockout mice, but remained decreased in wild-type mice. Radioligand binding was unaltered on retest scanning. Our results suggest that the prolonged decrease of radioligand binding after amphetamine is mainly due to internalization of the D(2) receptor rather than dopamine displacement. In addition, this study demonstrates the utility of small animal PET to study receptor trafficking in vivo in genetically modified mice.
[show abstract][hide abstract] ABSTRACT: Recent applications of micro-reactor (microfluidics) technology to radiofluorination chemistry within our laboratory are presented, based on use of either a simple T-shaped glass micro-reactor or a more advanced microfluidics instrument. The topics include reaction optimization and radioligand production, in particular the study of the radiofluorination of diaryliodonium salts, [(18)F]fluoride ion exchange with xenon difluoride(,) esterification with [(18)F]2-fluoroethyl tosylate, and the syntheses of [(18)F]fallypride, [(18)F]FBR and [(18)F]SL702 from [(18)F]fluoride ion.
Journal of Labelled Compounds and Radiopharmaceuticals 05/2010; 53(5-6):234-238. · 1.24 Impact Factor
[show abstract][hide abstract] ABSTRACT: A microreactor was applied to produce ortho-substituted [(18)F]fluoroarenes from the reactions of cyclotron-produced [(18)F]fluoride ion (t(1/2) = 109.7 min) with diaryliodonium salts. The microreactor provided a very convenient means for running sequential reactions rapidly with small amounts of reagents under well-controlled conditions, thereby allowing reaction kinetics to be followed and Arrhenius activation energies (E(a)) to be measured. Prepared symmetrical iodonium chlorides (Ar(2)I(+)Cl(-)) rapidly (<4 min) gave moderate (Ar = 2-MeOC(6)H(4), 51%) to high (Ar = Ph or 2-MeC(6)H(4), 85%) decay-corrected radiochemical yields (RCYs) of a single radioactive product (Ar(18)F). Reaction velocity with respect to Ar group was 2-MeOC(6)H(4) < Ph < 2-MeC(6)H(4). Activation energies were in the range 18-28 kcal/mol. Prepared unsymmetrical salts (e.g., 2-RC(6)H(4)I(+)2'-R'C(6)H(4)X(-); X = Cl or OTs) also rapidly gave two products (2-RC(6)H(4)(18)F and 2-R'C(6)H(4)(18)F) in generally high total RCYs (79-93%). Selectivity for product [(18)F]fluoroarene was controlled by the nature of the ortho substituents. The power of ortho substituents to impart an ortho-effect was in the following order:, 2,6-di-Me > 2,4,6-tri-Me > Br > Me > Et approximately (i)Pr > H > OMe. For (2-methyphenyl)(phenyl)iodonium chloride, the time-course of reaction product selectivity was constant and consistent with the operation of the Curtin-Hammett principle. These results will aid in the design of diaryliodonium salt precursors to (18)F-labeled tracers for molecular imaging.
The Journal of Organic Chemistry 04/2010; 75(10):3332-8. · 4.56 Impact Factor
[show abstract][hide abstract] ABSTRACT: In vitro and ex vivo measurements have shown that the binding of the selective high-affinity agonist, S14506 (1-[2-(4-fluorobenzoylamino)ethyl]-4-(7-methoxy-naphthyl)piperazine), to 5-HT(1A) receptors, is similar in affinity (K(d) = 0.79 nM) and extent (B(max)) to that of the antagonist, WAY 100635. We aimed to test whether S14506, labeled with a positron-emitter, might serve as a radioligand for imaging brain 5-HT(1A) receptors in vivo with positron emission tomography (PET). Here we evaluated [(11)C]S14506 and [(18)F]S14506 in rat and rhesus monkey in vivo. After intravenous administration of [(11)C]S14506 into rat, radioactivity entered brain, reaching 210% SUV at 2 min. Radioactivity uptake into brain was higher (~ 350% SUV) in rats pre-treated with the P-glycoprotein (P-gp) inhibitor, cyclosporin A. In rhesus monkey, peak brain uptake of radioactivity after administration of [(11)C]S14506 or [(18)F]S14506 was also moderate and for [(11)C]S14506 increased from ~ 170% SUV after 7 min, to 240% SUV in a monkey pre-treated with the P-gp inhibitor, tariquidar. The ratios of radioactivity in 5-HT(1A) receptor-rich regions, such as cingulate or hippocampus to that in receptor-poor cerebellum reached between 1.35 and 1.5 at 60 min for both [(11)C]S14506 and [(18)F]S14506. [(11)C]S14506 gave one major polar radiometabolite in monkey plasma, and [(18)F]S14506 gave three and two more polar radiometabolites in rat and monkey plasma, respectively. The rat radiometabolites of [(18)F]S14506 did not accumulate in brain. [(18)F]S14506 was not radiodefluorinated in monkey. Thus, despite high-affinity and lack of troublesome brain radiometabolites, both [(11)C]S14506 and [(18)F]S14506 were ineffective for imaging rat or monkey brain 5-HT(1A) receptors in vivo, even under P-gp inhibited conditions. Explanations for the failure of these radioligands are offered.
[show abstract][hide abstract] ABSTRACT: A series of arylsulfonate nucleophile assisting leaving groups (NALGs) were prepared in which the metal chelating unit is attached to the aryl ring via an ether linker. These NALGs exhibited significant rate enhancements in halogenation reactions using metal halides. Studies with a NALG containing a macrocyclic ether unit suggest that rate enhancements of these nucleophilic halogenation reactions are facilitated by stabilization of charge in the transition state rather than through strong precomplexation with metal cation. In several cases, a primary substrate containing one of the new leaving groups rivaled or surpassed the reactivity of triflates when exposed to nucleophile but was otherwise highly stable and isolable. These and previously disclosed chelating leaving groups were used in (18)F-fluorination reactions using no-carrier-added [18F]fluoride ion (t(1/2) = 109.7 min, beta+ = 97%) in CH3CN. Under microwave irradiation and without the assistance of a cryptand, such as K2.2.2, primary substrates with select NALGs led to a substantial improvement (2-3-fold) in radiofluorination yields over traditional leaving groups.
The Journal of Organic Chemistry 08/2009; 74(15):5290-6. · 4.56 Impact Factor
[show abstract][hide abstract] ABSTRACT: Elevated levels of peripheral benzodiazepine receptors (PBR) are associated with activated microglia in their response to inflammation. Hence, PBR imaging in vivo is valuable for investigating brain inflammatory conditions. Sensitive, easily prepared, and readily available radioligands for imaging with positron emission tomography (PET) are desirable for this purpose. We describe a new 18F-labeled PBR radioligand, namely [18F]N-fluoroacetyl-N-(2,5-dimethoxybenzyl)-2-phenoxyaniline ([18F]9). [18F]9 was produced easily through a single and highly efficient step, the reaction of [18F]fluoride ion with the corresponding bromo precursor, 8. Ligand 9 exhibited high affinity for PBR in vitro. PET showed that [18F]9 was avidly taken into monkey brain and gave a high ratio of PBR-specific to nonspecific binding. [18F]9 was devoid of defluorination in rat and monkey and gave predominantly polar radiometabolite(s). In rat, a low level radiometabolite of intermediate lipophilicity was identified as [18F]2-fluoro-N-(2-phenoxyphenyl)acetamide ([18F]11). [18F]9 is a promising radioligand for future imaging of PBR in living human brain.
Journal of Medicinal Chemistry 02/2009; 52(3):688-99. · 5.61 Impact Factor