Norman Koglin

Exploratory clinical trial of (4S)-4-(3-[18F]fluoropropyl)-L-glutamate for imaging xC- transporter using positron emission tomography in patients with non-small cell lung or breast cancer.

Purpose: (4S)-4-(3-[18F]fluoropropyl)-l-glutamate (BAY 94-9392, alias [18F]FSPG) is a new tracer to image xC− transporter activity with positron emission tomography (PET). We aimed to explore the tumor detection rate of [18F]FSPG in patients relative to 2-[18F]fluoro-2-deoxyglucose ([18F]FDG). The correlation of [18F]FSPG uptake with immunohistochemical expression of xC− transporter and CD44, which stabilizes the xCT subunit of system xC−, was also analyzed. Experimental Design: Patients with non–small cell lung cancer (NSCLC, n = 10) or breast cancer (n = 5) who had a positive [18F]FDG uptake were included in this exploratory study. PET images were acquired following injection of approximately 300 MBq [18F]FSPG. Immunohistochemistry was done using xCT- and CD44-specific antibody. Results: [18F]FSPG PET showed high uptake in the kidney and pancreas with rapid blood clearance. [18F]FSPG identified all 10 NSCLC and three of the five breast cancer lesions that were confirmed by pathology. [18F]FSPG detected 59 of 67 (88%) [18F]FDG lesions in NSCLC, and 30 of 73 (41%) in breast cancer. Seven lesions were additionally detected only on [18F]FSPG in NSCLC. The tumor-to-blood pool standardized uptake value (SUV) ratio was not significantly different from that of [18F]FDG in NSCLC; however, in breast cancer, it was significantly lower (P < 0.05). The maximum SUV of [18F]FSPG correlated significantly with the intensity of immunohistochemical staining of xC− transporter and CD44 (P < 0.01). Conclusions: [18F]FSPG seems to be a promising tracer with a relatively high cancer detection rate in patients with NSCLC. [18F]FSPG PET may assess xC− transporter activity in patients with cancer. Clin Cancer Res; 18(19); 5427–37. ©2012 AACR.

New protein deposition tracers in the pipeline

Traditional nuclear medicine ligands were designed to target cellular receptors or transporters with a binding pocket and a defined structure–activity relationship. More recently, tracers have been developed to target pathological protein aggregations, which have less well-defined structure–activity relationships. Aggregations of proteins such as tau, α-synuclein, and β-amyloid (Aβ) have been identified in neurodegenerative diseases, including Alzheimer’s disease (AD) and other dementias, and Parkinson’s disease (PD). Indeed, Aβ deposition is a hallmark of AD, and detection methods have evolved from coloured dyes to modern 18F-labelled positron emission tomography (PET) tracers. Such tracers are becoming increasingly established in routine clinical practice for evaluation of Aβ neuritic plaque density in the brains of adults who are being evaluated for AD and other causes of cognitive impairment. While similar in structure, there are key differences between the available compounds in terms of dosing/dosimetry, pharmacokinetics, and interpretation of visual reads. In the future, quantification of Aβ-PET may further improve its utility. Tracers are now being developed for evaluation of tau protein, which is associated with decreased cognitive function and neurodegenerative changes in AD, and is implicated in the pathogenesis of other neurodegenerative diseases. While no compound has yet been approved for tau imaging in clinical use, it is a very active area of research. Development of tau tracers comprises in-depth characterisation of existing radiotracers, clinical validation, a better understanding of uptake patterns, test-retest/dosimetry data, and neuropathological correlations with PET. Tau imaging may allow early, more accurate diagnosis, and monitoring of disease progression, in a range of conditions. Another marker for which imaging modalities are needed is α-synuclein, which has potential for conditions including PD and dementia with Lewy bodies. Efforts to develop a suitable tracer are ongoing, but are still in their infancy. In conclusion, several PET tracers for detection of pathological protein depositions are now available for clinical use, particularly PET tracers that bind to Aβ plaques. Tau-PET tracers are currently in clinical development, and α-synuclein protein deposition tracers are at early stage of research. These tracers will continue to change our understanding of complex disease processes.

[18F]Fluoropyruvate: radiosynthesis and initial biological evaluation

The radiosynthesis of [(18)F]fluoropyruvate was investigated using numerous precursors were synthesized from ethyl 2,2-diethoxy-3-hydroxypropanoate (5) containing different leaving groups: mesylate, tosylate, triflate, and nonaflate. These precursors were evaluated for [(18)F]fluoride incorporation with triflate being superior. The subsequent hydrolysis step was investigated, and an acidic hydrolysis was optimized. After establishing suitable purification and formulation methods, the [(18)F]fluoropyruvate could be isolated in ca. 50% d.c. yield. The [(18)F]fluoropyruvate was evaluated in vitro for its uptake into tumor cells using adenocarcinomic human alveolar basal epithelial cells (A549) and unfortunately showed an uptake of approximately 0.1% of the applied dose per 100,000 cells after 30 min. Initial pharmacokinetic properties were assessed in vivo using nude mice showed a high degree of bone uptake from defluorination, which will limit its potential as an imaging agent for metabolic processes.

How to Identify Suitable Molecular Imaging Biomarkers

Molecular imaging biomarkers offer the unique possibility for earlier disease detection and better characterisation, as well as enabling of drug development. Addressing a relevant biomedical question, selecting the right target and the optimisation of ligand binding as well as pharmacokinetic parameters in preclinical models are the crucial steps before a suitable imaging agent can enter the clinical evaluation. General concepts for the selection of a suitable molecular imaging biomarker and preclinical aspects for target and ligand evaluation are presented and illustrated with examples. The clinical development of new imaging agents is discussed in light of the recently approved beta-amyloid plaque imaging agents and the diagnostic need in Alzheimer’s disease.

Abstract LB-227: Pilot clinical trials of FSPG (BAY 94-9392): An 18F-labeled glutamate derivative for PET imaging of system xC- activity in tumors

Proceedings: AACR 103rd Annual Meeting 2012‐‐ Mar 31‐Apr 4, 2012; Chicago, IL Visualizing adaptations of tumor metabolism by employing positron emission tomography (PET) with a specific imaging probe has the potential to better characterize tumors with higher sensitivity and specificity. The glutamate-cystine exchanger (system xC-, a SLC7A11/SLC3A2 heterodimer) is an important transporter in tumor cells and a promising new target for imaging. This exchanger transports the amino acids L-cystine and L-glutamate, thus providing a rate-limiting precursor for glutathione biosynthesis. This contributes to redox maintenance, which is especially important in tumor cells. Recently, it was reported that a splice variant of the stem cell marker CD44 stabilizes the xCT subunit of system xC- at the cell membrane and promotes its proper function. FSPG (BAY 94-9392) is an 18F-labeled glutamate derivative that provides specific visualization of system xC- activity with PET/CT. Through this multicenter pilot trial, FSPG has successfully been translated into cancer patients and shows favorable pharmacokinetics and an absence of adverse events. A favorable biodistribution pattern previously seen in mice was also confirmed in these patients with different tumor types. A high lesion detection rate (80-100%) was found in lung cancer (n=10), hepatocellular cancer (n=5), head and neck cancer (n=5), and colorectal cancer (n=5) patients who also had positive FDG-PET/CT scans. Importantly, very low background uptake in healthy brain parenchyma enabled the detection of high-grade brain cancers as well as small brain metastases and skull base lesions (n=4) not well seen with FDG-PET. Primary breast tumors (n=4) and a low-grade brain tumor, however, showed minimal uptake of FSPG. Immunohistochemical (IHC) analyses on these subjects’ pathology samples showed significant correlation between FSPG uptake and protein expression of both the xCT subunit and CD44. In breast tumor samples specifically, IHC showed that absence of CD44 results in low signal from FSPG-PET even if xCT is present, indicating the importance of CD44 co-expression for system xC- function. Future clinical studies will further explore the potential of this compound in additional tumor types and expand upon the initial findings in a larger number of subjects. Here we present the first clinical examples demonstrating the ability of the tracer to measure system xC- activity in patients. FSPG-PET/CT may provide improved characterization and sensitivity of tumor detection in patients and further improve the understanding of the role of system xC- in cancer biology. It could be a valuable tool for improved tumor classification in patients, linking uptake of the tracer with malignancy and cancer stem cell properties of tumors. Furthermore, the ability to noninvasively probe this important pathway in humans may allow for more rational treatment planning, patient stratification and better development of new drugs. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-227. doi:1538-7445.AM2012-LB-227

Abstract 2800: BAY 94-9392 – A new derivative of L-glutamate for tumor specific PET imaging

There is a high demand for a tumor specific imaging, staging and therapy monitoring by PET. Currently, FDG (2-[F-18]fluoro-2-deoxyglucose) is the commonly used agent for tumor detection and staging. However, this method cannot differentiate between tumor and inflammatory lesions. In addition, FDG does not accumulate in tumors with low glycolytic activities, such as prostate cancer. It is not optimal for specific detection of brain tumors and metastases due to a high background signal resulting from healthy brain metabolism. The amino acids L-glutamine and L-glutamate as well as their corresponding transporters play a key role in the adapted intermediary metabolism of tumors. We synthesized and evaluated a new PET imaging agent BAY 94-9392, a derivative of L-glutamate. This agent was successfully used for tumor detection with high sensitivity and specificity as demonstrated in various preclinical models. Methods: The radiosynthesis of (4S)-4-(3-[F-18]fluoropropyl)-L-glutamate (BAY 94-9392) was established on a commercial synthesis module. BAY 94-9392 was investigated in a variety of tumor cell lines as well as tumor models for uptake, biodistribution, mode of action, and its potential to differentiate between tumor versus inflammation. Results: BAY 94-9392 is specifically transported by the cystine/glutamate exchanger (xCT, SLC7A11) and it is not metabolized. Compared to FDG, the agent displayed a high accumulation rate in a panel of tumor cells. PET imaging with excellent tumor visualization and high tumor to background ratios was achieved in a variety of preclinical tumor models. BAY 94-9392 did not accumulate in inflammatory lesions, whereas FDG shows a strong uptake in this turpentine oil induced and histopathologically confirmed rat model. Conclusions: BAY 94-9392 is a new, promising tumor specific PET tracer. Both preclinical and clinical studies are in progress for further characterization. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2800. doi:10.1158/1538-7445.AM2011-2800