Imaging the 5-HT(1A) receptors with PET: WAY-100635 and analogues

This paper summarizes our work on WAY-100635 and analogues, labeled either with carbon-11 or fluorine-18, as potential radioligands for the 5-hydroxytryptamine(1A) (5-HT(1A)) neuroreceptors. Other facets of our work include: (1) human studies with [O-methyl-(11)C]WAY-100634, the major radioactive metabolite of [O-methyl-(11)C]WAY-100635, and with [(11)C]CPC 222; (2) use of a human liver metabolism model to screen in vitro potential metabolic problems in humans; (3) modification of the "dry method" to prepare [carbonyl-(11)C]WAY-100635; and (4) validation studies in humans with [carbonyl-(11)C]WAY-100635.     

PET evaluation of [(18)F]FCWAY, an analog of the 5-HT(1A) receptor antagonist, WAY-100635

We synthesized [(18)F]FCWAY, an analog of [carbonyl-(11)C]WAY-100635 ?[(11)C]N-(2-(1-(4-(2-methoxyphenyl)-piperazinyl)ethyl))-N-(2-(pyridi nyl))cyclohexanecarboxamide?, by replacing the cyclohexanecarbonyl group acid with a trans-4-fluorocyclohexanecarbonyl group (FC). Control and preblocking studies were performed in anesthetized monkeys. Plasma radioactive metabolite analysis showed the presence of [(18)F]FC and [(18)F]fluoride. Tissue time-radioactivity curves were corrected for metabolite contamination based on separate positron-emission tomography studies of these two labeled metabolites. Analysis using a two-tissue compartment model gave distribution volume (V) estimates (mL/mL) ranging from 33 in frontal cortex to 4 in cerebellum. Preblocking data showed uniform V of 2-3 mL/mL. These studies demonstrate that [(18)F]FCWAY has very similar kinetic characteristics to [(11)C]WAY-100635.     

Use of PET and the radioligand [carbonyl-(11)C]WAY-100635 in psychotropic drug development

Positron-emission tomography (PET) provides potential in neuropsychiatric drug development by expanding knowledge of drug action in the living human brain and reducing time consumption and costs. The 5-hydroxytryptamine(1A) (5-HT(1A)) receptor is of central interest as a target for the treatment of anxiety, depression, and schizophrenia. Research on the clinical significance of the 5-HT(1A) receptor now benefits from the highly selective radioligand [carbonyl-(11)C]WAY-100635 (WAY) for quantitative determination of 5-HT(1A) receptors in the primate and human brain in vivo using PET. In this paper, three studies are reviewed to demonstrate the suitability of WAY as radioligand for quantification of central 5-HT(1A) receptors in brain and as an applicable tool for drug development. In the first study a monkey model was used to characterize WAY binding. It was confirmed that the reference ligand 8-OH-DPAT and psychoactive drugs such as buspirone and pindolol occupies 5-HT(1A) receptors in the primate brain. Pindolol is an beta-adrenoreceptor antagonist with a high affinity to 5-HT(1A) receptors. This drug has been suggested in combination with selective serotonin reuptake inhibitors for the treatment of depression and was given to healthy males in the second study. Pindolol induced a marked inhibition of central 5-HT(1A) receptors as calculated by the ratio-analysis method and simplified reference tissue model, 2 h after administration of 10 mg as a single oral dose. This observation suggests that pindolol may have a role for the suggested potentiation of selective serotonin reuptake inhibitor treatment of depression. The third study was on robalzotan (NAD-299), a recently developed 5-HT(1A) receptor antagonist and putative drug with implications for the treatment of depression. In the cynomolgus monkey brain, robalzotan in the dose range 2-100 microg/kg IV occupied 5-HT(1A) receptors in a dose-dependent and saturable manner with a maximal calculated occupancy of 70-80%. The relationship between robalzotan plasma concentration and 5-HT(1A) receptor occupancy could be described by a hyperbolic function that was used to guide the selection of appropriate doses in man. In a subsequent PET study of robalzotan binding to 5-HT(1A) receptors in the living human brain, similar results have been replicated recently. These studies reviewed here illustrate and corroborate that quantitative neuroimaging of receptor binding has potential for the evaluation and dose finding of new central nervous system drugs.     

A retrospect on the discovery of WAY-100635 and the prospect for improved 5-HT(1A) receptor PET radioligands

Abstract. An outline is given of the developments that led to the identification of [O-methyl-(11)C]WAY-100635 (4) as the first useful PET ligand for imaging serotonin(1A) (5-HT(1A)) receptors in the living human brain. Recent attempts to develop 5-HT(1A) receptor radioligands superior to 4 are reviewed, and [carbonyl-(11)C]WAY-100635 (6) has been shown to be the best currently available radioligand for human studies. Of other (11)C-radiolabelled compounds, [O-methyl-(11)C](R,S)-CPC-222 (9), DWAY (8), and [(11)C]NAD-299 (14) all demonstrate specific binding to 5-HT(1A) receptors in animals and warrant further expedited studies in humans. The trans-fluorocyclohexane, 12, and fluorobenzene, [(18)F]p-MPPF 13, are highlighted as examples of promising (18)F-labelled ligands.     

The PET radioligand [carbonyl-(11)C]desmethyl-WAY-100635 binds to 5-HT(1A) receptors and provides a higher radioactive signal than [carbonyl-(11)C]WAY-100635 in the human brain.

5-Hydroxytryptamine (serotonin)-1A (5-HT(1A)) receptors are of key interest in research on the pathophysiology and treatment of psychiatric disorders. The PET radioligand [carbonyl-(11)C]WAY-100635 ((11)C-WAY), where WAY-100635 is (3)H-(N-(2-(1-(4-(2-methoxyphenyl)-1-piperazinyl)ethyl)-N-(2-pyridyl) cyclohexane-carboxamide, is commonly used for quantitation of 5-HT(1A) receptors in the human brain. The aim of this PET study was to compare (11)C-WAY with the putative metabolite and selective radioligand [carbonyl-(11)C]desmethyl-WAY-100635 ((11)C-DWAY). METHODS: A PET examination was performed on each of 5 healthy male volunteers after intravenous injection of (11)C-WAY and (11)C-DWAY on separate occasions. Radioactive metabolites in plasma were determined with high-performance liquid chromatography. The plasma metabolite--corrected input function was used in a kinetic compartment analysis. The simplified reference tissue model and peak equilibrium method, using the cerebellum as reference region, was applied for comparison of data. RESULTS: For both radioligands, the highest radioactivity was observed in the neocortex and the raphe nuclei, whereas radioactivity was low in the cerebellum. The regional binding potentials were similar for the 2 radioligands. The brain uptake was more than 2-fold higher for (11)C-DWAY than for (11)C-WAY, in part because of higher delivery (first-order rate constant K(1), 0.38 vs. 0.16). The time--activity curves were well described by a 3-compartment model for all regions, whereas uptake in the cerebellum could not be described by a 2-compartment model, supporting the existence of kinetically distinguishable nonspecific binding in the cerebellum or radioactive metabolites in the brain for both radioligands. Both radioligands were rapidly metabolized, and <10% of the radioactivity in plasma represented unchanged (11)C-WAY or (11)C-DWAY at 10 min after injection. The metabolic pattern was similar for both radioligands, with the formation of radiolabeled cyclohexanecarboxylic acid and more polar components. For (11)C-WAY, small amounts of an additional labeled metabolite comigrated with reference desmethyl-WAY-100635. CONCLUSION: The advantages of (11)C-DWAY over (11)C-WAY for research on central 5-HT(1A) receptors is supported by a significantly higher radioactivity signal at equipotent doses, providing improved imaging statistics and advantages in biomathematic modeling and the preclusion of (11)C-DWAY as a metabolite interfering with PET measurements.     

Quantification of [Carbonyl-(11)C]WAY-100635 binding: considerations on the cerebellum

The specific binding of [carbonyl-(11)C]WAY-100635 to 5-hydroxytryptamine(1A) receptors was quantitated using different kinetic models. Ten healthy subjects were studied using a GE Advance PET scanner. We suggest that tissue heterogeneity explains part of the nonspecific binding seen in cerebellum, which leads to an underestimation of binding potential with reference tissue methods. The nonlinear three-compartmental model with metabolite-corrected plasma input provides accurate estimates of receptor binding because of the favorable kinetics of tracer in the brain and circulation.     

Drug action at the 5-HT(1A) receptor in vivo: autoreceptor and postsynaptic receptor occupancy examined with PET and [carbonyl-(11)C]WAY-100635

Serotonin(1A) (5-HT(1A)) receptors have been implicated in the pathophysiology and treatment of anxiety and depression and are a target for novel drug development. In this qualitative study, positron emission tomography (PET) and [carbonyl-(11)C]WAY-100635 were used to assess 5-HT(1A) autoreceptor and postsynaptic receptor occupancy in man in vivo by five different compounds with nanomolar affinity for this site. Occupancy by pindolol, penbutolol, buspirone, EMD 68843, and S 15535 was compared to test-retest data from 10 healthy volunteers. All drugs, apart from buspirone, displayed occupancy at the 5-HT(1A) receptor site. Pindolol demonstrated a preferential occupancy at the autoreceptor compared to the postsynaptic receptor over a plasma range of about 10-20 ng/mL. Differential occupancy may be an important component of novel drug action. The level of autoreceptor or postsynaptic occupancy needed to achieve significant physiological effects is not known, although it is of note that none of the drugs in this study achieved occupancies beyond 60%. Overall this study demonstrates the utility of PET in aiding novel drug development.     

Synthesis of S-([18F]fluoromethyl)-(+)-McN5652 as a potential PET radioligand for the serotonin transporter

The present study describes the synthesis of the [18F]fluoromethyl analogue of (+)-McN5652 ([18F]FMe-McN) as a new potential tracer for the serotonin transporter. In vitro binding studies have shown that FMe-McN displays only slightly lower affinity for the serotonin transporter (K(i) = 2.3 +/- 0.1 nM) than (+)-McN5652 (K(i) = 0.72 +/- 0.2 nM). The radiofluorinated tracer [18F]FMe-McN was prepared by reaction of normethyl (+)-McN5652 with the fluoromethylation agent [18F]bromofluoromethane in an overall radiochemical yield of 5 +/- 1% (decay-corrected, related to [18F]fluoride) and with high specific radioactivity (200-2,000 GBq/micromol at the end of synthesis).     

Synthesis of fluorine-18-labeled ciprofloxacin for PET studies in humans

Ciprofloxacin (1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-(1-piperazinyl)-quinoline-3-carboxylic acid), a widely-prescribed antibiotic, was labeled with fluorine-18 with the aim to perform positron emission tomography studies in humans for pharmacokinetic measurements. Due to a lack of chemical activation of ciprofloxacin for a direct nucleophilic exchange reaction a novel two-step synthetic approach, which employed an activated 6-fluoro-7-chloro substituted precursor molecule, was developed. The radiosynthesis yielded, starting from 52.5 +/- 11.3 GBq of [(18)F]fluoride, 1.3 +/- 0.6 GBq (n = 13) [(18)F]ciprofloxacin ready for intravenous administration in about 130 min synthesis time. A series of analytical tests was performed in order to prove the identity of the radiolabeled compound and its suitability for human applications.     

The serotonin-1A receptor distribution in healthy men and women measured by PET and [carbonyl-11C]WAY-100635

Purpose  The higher prevalence rates of depression and anxiety disorders in women compared to men have been associated with sexual dimorphisms in the serotonergic system. The present positron emission tomography (PET) study investigated the influence of sex on the major inhibitory serotonergic receptor subtype, the serotonin-1A (5-HT_1A) receptor. Methods  Sixteen healthy women and 16 healthy men were measured using PET and the highly specific radioligand [carbonyl-¹¹C]WAY-100635. Effects of age or gonadal hormones were excluded by restricting the inclusion criteria to young adults and by controlling for menstrual cycle phase. The 5-HT_1A receptor BP_ND was quantified using (1) the ‘gold standard’ manual delineation approach with ten regions of interest (ROIs) and (2) a newly developed delineation method using a PET template normalized to the Montreal Neurologic Institute space with 45 ROIs based on automated anatomical labeling. Results  The 5-HT_1A receptor BP_ND was found equally distributed in men and women applying both the manual delineation method and the automated delineation approach. Women had lower mean BP_ND values in every region investigated, with a borderline significant sex difference in the hypothalamus (p = 0.012, uncorrected). There was a high intersubject variability of the 5-HT_1A receptor BP_ND within both sexes compared to the small mean differences between men and women. Conclusions  To conclude, when measured in the follicular phase, women do not differ from men in the 5-HT_1A receptor binding. To explain the higher prevalence of affective disorders in women, further studies are needed to evaluate the relationship between hormonal status and the 5-HT_1A receptor expression.     

Imaging infection and inflammation in children with (18)F-FDG PET and (18)F-FDG PET/CT

The goal of this article is to delineate indications for (18)F-FDG PET/CT pertaining to inflammation in the pediatric population, with emphasis on those that have been described in the literature. The limitations of (18)F-FDG PET/CT in this type of evaluation are also described, and the importance of using as low a dose as feasible is discussed. After reading this article, the reader should be able to list several clinical situations in which (18)F-FDG PET/CT may be appropriate, describe some limitations of (18)F-FDG PET/CT, and explain why dose is particularly important in the pediatric population.     

肺癌18F-FLT PET和18F-FDG PET的临床对比研究

目的 探讨18F-FLT PET在肺癌诊断和肿瘤增殖检测方面的应用价值.方法 对2005年9月-2008年10月解放军总医院收治的36例(男27例,女9例,年龄38～74岁)胸部CT疑似肺癌患者进行18F-FLT PET扫描,对同期收治的42例(男29例,女13例,年龄37～75岁)胸部CT疑似肺癌患者进行18F-FD...     

6-18F-多巴的制备及其大鼠体内分布研究

目的：制备PET显像剂6－^18F-多巴（6－^18F-DOPA)并研究其在大鼠体内的生物分布。方法：使用小型回旋加速器生产^18F离子,经亲核取代、手性相转移催化烷基化等4步反应,合成得到无载体的6-18F-DOPA。24只SD大鼠分为6组,每只经尾静脉注射1．11MBq 6-^18F-DOPA,经5,30,60,90,120和150min后分别处死,解剖,取有关组织、脏器称重并测定放射性计数。结果：6-^18F-DOPA合成的放化产率为3．8％－7．5％（衰变校正后）,合成时间100min,放化纯度大于99％。大鼠体内生物分布显示纹状体内有明显的放射性浓集,而大脑、皮质、小脑中放射性较低;纹状体／小脑放射性比值在60min达5．9;其他组织器官中的放射性快速消除,无明显浓集。结论：建立的6－^18F-DOPA合成方法有效可靠,6－^18F-DOPA主要分布在纹状体内。     

Synthesis and biodistribution of labelled p-iodo phentermine (IP), N,N,-dimethyl-p-iodo phentermine (IDMP) and N-isopropyl-p-iodo phentermine (IIP) in rats

p-Iodo-phentermine (IP) and two of its derivatives, N,N,-dimethyl-p-iodo-phentermine (IDMP) and N-isopropyl-p-iodo-phentermine (IIP) were synthesized and radiolabelled with iodine by isotopic exchange. They were evaluated as potential brain imaging agents and compared to IAMP biodistribution in rats did not show any superiority to IAMP.     

Preparation of (Re) Re-AEDP and its biodistribution studies

The synthesis of the Re (V) complex and preparation of ¹⁸⁸Re-AEDP are described using ¹⁸⁸Re which was obtained from the alumina-based ¹⁸⁸W/¹⁸⁸Re generator. Dependence of the radiolabeling yields of ¹⁸⁸Re-AEDP on reducing agent concentration, AEDP concentration, pH and addition of carrier was examined. In the case of optimum conditions, the radiolabeling yields of ¹⁸⁸Re-AEDP were 92–93% for carrier-free ¹⁸⁸Re and 95–98% for carrier-added ¹⁸⁸Re. The stability of ¹⁸⁸Re-AEDP at pH≈6 was studied and it is found that the carrier has a significant effect on the stability of ¹⁸⁸Re-AEDP. The biodistribution of carrier-free and carrier-added ¹⁸⁸Re-labelled compounds in rats was also measured. The results show that ¹⁸⁸Re (carrier-added)-AEDP is a potential bone palliation radiopharmaceutical due to its high skeletal uptake, rapid blood clearance and relatively low soft tissue absorption.     

Prevalence, challenges, and solutions for (18)F-FDG PET studies of obese patients: a technologist's perspective

Obesity has reached epidemic proportions in the United States; hence, it is frequently encountered in patients undergoing (18)F-FDG PET studies. The purpose of the current study was to present a technologist's perspective on the prevalence of obesity and the challenges and solutions in imaging obese patients in our PET facility. METHODS: From October 2002 to October 2003, whole-body (18)F-FDG PET was performed on 1,164 patients with a known or suspected malignancy. Images were acquired 45-60 min after (18)F-FDG injection (7.4 MBq [0.2 mCi]/kg, with a maximum of 925 MBq [25 mCi]) on a PET scanner using a 4-min emission and 3-min transmission time per bed position. A database was maintained of patient height and weight, and body mass index (BMI) was calculated. Patient obesity was classified as overweight (BMI > or = 25 kg/m(2)), obese (BMI > or = 30 kg/m(2)), or malignantly obese (BMI > or = 40 kg/m(2)). In addition, PET technologists recorded any problems and attempted solutions related to the patient weight. RESULTS: BMI calculations showed that 528 patients (45.4%) were overweight or obese (322 men and 206 women; mean age, 55 y). Of those, 201 (38%) were overweight, 270 (51%) were obese, and 57 (11%) were malignantly obese. Problems encountered in these patients included difficult intravenous access (15%), difficult patient positioning (10%), patient motion (7%), an incomplete study (emission only) (1%), and potentially higher radiation exposure to the technologist because of extra time spent near the patient. Attempted solutions included adjusting the schedule to allow more time per patient, adjusting the dose based on body weight, using varied positioning techniques, dividing the study to allow a respite between different image combinations, and dividing time spent with obese patients among the technologists involved. CONCLUSION: Excessive body weight and related problems have commonly been encountered in our PET facility. (18)F-FDG PET studies of obese patients represent an ongoing challenge, which requires patient-tailored solutions to avoid compromising image quality and risking higher radiation exposure to the technologists.     

Synthesis and evaluation of (18)F-labeled choline analogs as oncologic PET tracers.

Elevated levels of choline (trimethyl-2-hydroxyethylammonium) and choline kinase (CK) activity in neoplasms have motivated the development of positron-labeled choline analogs for noninvasive detection of cancer using PET. The aim of this study was to further evaluate [(18)F]fluorocholine (fluoromethyl-dimethyl-2-hydroxyethylammonium [FCH]) as an oncologic probe in comparison with several other closely related molecules. METHODS: FCH, [(18)F]fluoromethyl-methylethyl-2-hydroxyethylammonium (FMEC), [(18)F]fluoroethyl-dimethyl-2-hydroxyethylammonium (FEC), and [(18)F]fluoropropyl-dimethyl-2-hydroxyethylammonium (FPC) were synthesized through [(18)F]fluoroalkylation reactions. In vitro phosphorylation rates of the (18)F-labeled choline analogs and [methyl-(14)C]choline (CH) were studied using yeast CK. Several choline radiotracers were also evaluated in cultured PC-3 human prostate cancer cells. Data on chemical stability, radiation dosimetry, and toxicity of FCH were obtained. PET studies with FCH were performed on a patient with prostate cancer and a patient with a brain tumor. RESULTS: FCH and FMEC revealed in vitro phosphorylation by CK that was similar to that of choline, whereas rates of phosphorylation of FEC and FPC were 30% (P < 0.01) and 60% (P < 0.01) lower, respectively. Accumulations of FCH, CH, and FPC in cultured PC-3 cancer cells were comparable, whereas uptake of FEC was approximately one fifth that of FCH. Dosimetry estimates using FCH biodistribution data in mice indicated that the kidneys are radiation-dose-critical organs for FCH. PET images of a patient with recurrent prostate cancer showed uptake of FCH in the prostatic bed and in metastases to lymph nodes. FCH PET showed uptake in malignancies in a patient with metastatic breast cancer. PET revealed FCH uptake in biopsy-proven recurrent brain tumor with little confounding uptake by normal brain tissues. CONCLUSION: The fluoromethyl choline analog FCH may serve as a probe of choline uptake and phosphorylation in cancer cells, whereas fluoroethyl (FEC) and fluoropropyl (FPC) analogs appear to have relatively poorer biologic compatibility. Preliminary PET studies on patients with prostate cancer and with breast cancer and brain tumor support further studies to evaluate the usefulness of FCH as an oncologic probe.