Copper-64 based radiopharmaceuticals for brain tumors and hypoxia imaging.

INTRODUCTION: The most common and aggressive primary malignancy of the central nervous system is Glioblastoma that, as a wide range of malignant solid tumor, is characterized by extensive hypoxic regions. A great number of PET radiopharmaceuticals have been developed for the identification of hypoxia in solid tumors, among these, we find copper-based tracers. The aim of the current review paper was to provide an overview of radiocopper compounds applied for preclinical and clinical research in brain tumors and hypoxia imaging or therapy. EVIDENCE ACQUISITION: Copper offers a wide variety of isotopes, useful for nuclear medicine applications, but only 64Cu and 67Cu are under the spotlight of the scientific community since being good candidates for theranostic applications. Between the two, 64Cu availability and production cost have attracted more interest of the scientific community. EVIDENCE SYNTHESIS: In order to better understand the application of copper-bis thiosemicarbazones in hypoxia imaging, an overview of the role of hypoxia in cancer, existing non-imaging and imaging techniques for hypoxia identification and promising future avenues regarding hypoxia is necessary. Different proposed uptake mechanisms of [64Cu][Cu(ATSM)] inside the cell will be discussed and other 64Cu-based tracers for brain tumors described. CONCLUSIONS: Among radio copper compounds [64Cu][Cu(ATSM)] is the most studied radiopharmaceutical for imaging and treatment of brain tumors. Experimental evidence suggested that [64Cu][Cu(ATSM)] could be more appropriately considered as a marker of over-reduced intracellular state rather than a pure hypoxia agent. Moreover, preliminary clinical data suggested that [64Cu]CuCl2 can be a potentially useful diagnostic agent for malignancies of the central nervous system (CNS).

IAEA contribution to the development of 64Cu radiopharmaceuticals for theranostic applications.

Copper-64 is a very attractive radioisotope with unique nuclear properties that allow using it as both a diagnostic and therapeutic agent, thus providing an almost ideal example of a theranostic radionuclide. A characteristic of Cu-64 stems from the intrinsic biological nature of copper ions that play a fundamental role in a large number of cellular processes. Cu-64 is a radionuclide that reflects the natural biochemical pathways of Cu-64 ions, therefore, can be exploited for the detection and therapy of certain malignancies and metabolic diseases. Beside these applications of Cu-64 ions, this radionuclide can be also used for radiolabelling bifunctional chelators carrying a variety of pharmacophores for targeting different biological substrates. These include peptide-based substrates and immunoconjugates as well as small-molecule bioactive moieties. Fueled by the growing interest of Member States (MS) belonging to the International Atomic Energy Agency (IAEA) community, a dedicated Coordinated Research Project (CRP) was initiated in 2016, which recruited thirteen participating MS from four continents. Research activities and collaborations between the participating countries allowed for collection of an impressive series of results, particularly on the production, preclinical evaluation and, in a few cases, clinical evaluation of various 64Cu-radiopharmaceuticals that may have potential impact on future development of the field. Since this CRP was finalized at the beginning of 2020, this short review summarizes outcomes, outputs and results of this project with the purpose to propagate to other MS and to the whole scientific community, some of the most recent achievements on this novel class of theranostic 64Cu-pharmaceuticals.

Development of (153) Sm-folate-polyethyleneimine-conjugated chitosan nanoparticles for targeted therapy.

The aim of this study was to develop biocompatible, water-soluble (153) Sm-labeled chitosan nanoparticles (NPs) containing folate and polyethyleneimine functionalities i.e. chitosan-graft-PEI-folate (CHI-DTPA-g-PEI-FA), suitable for targeted therapy. The physicochemical properties of the obtained NPs were characterized by dynamic light-scattering analysis for their mean size, size distribution, and zeta potential; scanning electron microscopy for surface morphology; and (1) H-NMR, FT-IR analyses for molecular dispersity of folate in the NPs. NPs were spherical with mean diameter below 250 nm, polydispersity of below 0.15, and positive zeta potential values. The NP complex ((153) Sm-CHI-DTPA-g-PEI-FA) was stable at 25 °C (6-8 h, >90% radiochemical purity, instant thin layer chromatography (ITLC)). Binding studies using fluorescent NPs for internalization also demonstrated significant uptake in MCF-7 cells. MCF-7 cell internalization was significantly greater for 4T1. In blocking studies, both MCF-7 and 4T1 cell lines demonstrated specific folate receptor (FR) binding (decreasing 45%). In vivo biodistribution studies indicated major excretion of NPs metabolites and/or free (153) Sm through the kidneys. The preliminary imaging studies in 4T1 tumor-bearing mice showed minor uptake up to 96 h. The present folic acid that functionalized chitosan NP is a candidate material for folate receptor therapy.

Potential Theranostic Boron Neutron Capture Therapy Agents as Multimodal Radiopharmaceuticals.

Boron neutron capture therapy (BNCT) has been extant for decades and continues to be practiced in many centers around the globe. Most of the active clinical trials utilize boronophenylalanine as the drug containing boron atoms. The important aspect that has been added to the BNCT practice is the use of an F-18 radiolabeled analog for ascertaining targeting and monitoring follow-up studies. The recent widespread application of therapeutic radiopharmaceuticals, especially peptides (somatostatin analogs), prostate-specific antigen-binding ligands, or immunomolecules, offers the ambit for invention of new tumor-specific BNCT agents, especially for BNCT-susceptible tumors, that is, locoregional cancers such as head and neck cancer. Such BNCT agents, when radiolabeled, can enable simultaneous imaging and/or therapeutic applications (depending on the radionuclide used) through multimodal approaches. Development of boron-rich moieties such as sodium borocaptate and neutral carboranes combined with tumor-targeting moieties can lead to a new horizon in BNCT. The review covers various aspects of drug design, tumor targeting, and possible future radiopharmaceutical development for multimodal theranostic application in humans.

Current status on cyclotron facilities and related infrastructure supporting PET applications in Latin America and the Caribbean.

This review presents the results of a survey conducted by the International Atomic Energy Agency on cyclotrons and related infrastructure used for radionuclide and radiopharmaceutical production which are supporting PET imaging applications in Latin America and the Caribbean region.

IAEA Contribution to Nanosized Targeted Radiopharmaceuticals for Drug Delivery.

The rapidly growing interest in the application of nanoscience in the future design of radiopharmaceuticals and the development of nanosized radiopharmaceuticals in the late 2000's, resulted in the creation of a Coordinated Research Project (CRP) by the International Atomic Energy Agency (IAEA) in 2014. This CRP entitled 'Nanosized delivery systems for radiopharmaceuticals' involved a team of expert scientist from various member states. This team of scientists worked on a number of cutting-edge areas of nanoscience with a focus on developing well-defined, highly effective and site-specific delivery systems of radiopharmaceuticals. Specifically, focus areas of various teams of scientists comprised of the development of nanoparticles (NPs) based on metals, polymers, and gels, and their conjugation/encapsulation or decoration with various tumor avid ligands such as peptides, folates, and small molecule phytochemicals. The research and development efforts also comprised of developing optimum radiolabeling methods of various nano vectors using diagnostic and therapeutic radionuclides including Tc-99m, Ga-68, Lu-177 and Au-198. Concerted efforts of teams of scientists within this CRP has resulted in the development of various protocols and guidelines on delivery systems of nanoradiopharmaceuticals, training of numerous graduate students/post-doctoral fellows and publications in peer reviewed journals while establishing numerous productive scientific networks in various participating member states. Some of the innovative nanoconstructs were chosen for further preclinical applications-all aimed at ultimate clinical translation for treating human cancer patients. This review article summarizes outcomes of this major international scientific endeavor.

Highlight selection of radiochemistry and radiopharmacy developments by editorial board.

BACKGROUND: The Editorial Board of EJNMMI Radiopharmacy and Chemistry releases a biyearly highlight commentary to update the readership on trends in the field of radiopharmaceutical development. RESULTS: This commentary of highlights has resulted in 21 different topics selected by each member of the Editorial Board addressing a variety of aspects ranging from novel radiochemistry to first in man application of novel radiopharmaceuticals. Also the first contribution in relation to MRI-agents is included. CONCLUSIONS: Trends in (radio)chemistry and radiopharmacy are highlighted demonstrating the progress in the research field being the scope of EJNMMI Radiopharmacy and Chemistry.

Production and Supply of α-Particle-Emitting Radionuclides for Targeted α-Therapy.

Encouraging results from targeted α-therapy have received significant attention from academia and industry. However, the limited availability of suitable radionuclides has hampered widespread translation and application. In the present review, we discuss the most promising candidates for clinical application and the state of the art of their production and supply. In this review, along with 2 forthcoming reviews on chelation and clinical application of α-emitting radionuclides, The Journal of Nuclear Medicine will provide a comprehensive assessment of the field.

IAEA Activities on 67Cu, 186Re, 47Sc Theranostic Radionuclides and Radiopharmaceuticals.

Despite interesting properties, the use of 67Cu, 186Re and 47Sc theranostic radionuclides in preclinical studies and clinical trials is curtailed by their limited availability due to a lack of widely established production methods. An IAEA Coordinated Research Project (CRP) was initiated to identify important technical issues related to the production and quality control of these emerging radionuclides and related radiopharmaceuticals, based on the request from IAEA Member States. The international team worked on targetry, separation, quality control and radiopharmaceutical aspects of the radionuclides obtained from research reactors and cyclotrons leading to preparation of a standard recommendations for all Member States. The CRP was initiated in 2016 with fourteen participants from thirteen Member States from four continents. Extraordinary results on the production, quality control and preclinical evaluation of selected radionuclides were reported in this project that was finalized in 2020. The outcomes, outputs and results of this project achieved by participating Member States are described in this minireview.

Cyclotron production of no carrier added 186gRe radionuclide for theranostic applications.

186gRe (T1/2 = 3.7183 d, E(ß-)mean = 346.7 keV, I(ß-)mean = 92.59%), a mixed beta and γ-emitter shows great potential for use in theranostic applications. The dominant 185Re(n,γ) route, via use of a nuclear reactor, provides 186gRe in carrier added form with low specific activity, while cyclotrons offer no carrier-added (NCA) high specific activity production of 186gRe. However, to be able to select the best possible nuclear reaction and to optimize the production route via the use of a cyclotron, information on the excitation function for the reaction of interest as well as for the competing reactions is necessary. Accordingly, we have conducted a detailed study of the excitation functions for natW(d, x) reactions in seeking optimized parameters for the NCA production of 186gRe. Noting a discrepancy among the experimental data, we made an evaluation of the available literature, finally selecting optimum parameters for the production of 186gRe via the 186W(d,2n)186Re reaction. These beam parameters were then used for batch production of 186gRe by irradiating an enriched 186W metallic powder target, followed by a subsequent automated chemical separation process. The preliminary results show 98.1% radionuclidic purity of 186gRe at 8 h subsequent to the End of Bombardment (EOB), offering the potential for use in clinical applications.

Preclinical studies of [61Cu]ATSM as a PET radiopharmaceutical for fibrosarcoma imaging.

[61Cu]diacetyl-bis(N4-methylthiosemicarbazone) ([61Cu] ATSM) was prepared using in house-made diacetyl-bis(N4-methylthiosemicarbazone) (ATSM) ligand and [61Cu]CuCl2 produced via the natZn(p, x)61Cu (180 muA proton irradiation, 22 MeV, 3.2 h) and purified by a ion chromatography method. [61Cu]ATSM radiochemical purity was >98 %, as shown by HPLC and RTLC methods. [61Cu]ATSM was administered into normal and tumor bearing rodents for up to 210 minutes, followed by biodistribution and co-incidence imaging studies. Significant tumor/non-tumor accumulation was observed either by animal sacrification or imaging. [61Cu]ATSM is a positron emission tomography (PET) radiotracer for tumor hypoxia imaging.

The development of radiogallium-acetylacetonate bis(thiosemicarbazone) complex for tumour imaging.

BACKGROUND: Various radiometal complexes have been developed for tumour imaging, especially Ga-68 tracer. In this work, the development of a radiogallium bis(thiosemicarbazone) complex has been reported. MATERIAL AND METHODS: [(67)Ga]acetylacetonate bis(thio-semicarbazone) complex ([(67)Ga]AATS) was prepared starting with [(67)Ga]Gallium acetate and freshly prepared acetylacetonate bis(thiosemicarbazone) (AATS) for 30 min at 90 degreesC. The partition co-efficient and stability of the tracer was determined in final solution (25 degreesC) and the presence of human serum (37 degreesC) for up to 24 hours. The biodistribution of the labelled compound in wild-type and fibrosarcoma-bearing rodents were determined for up to 72 hours. RESULTS: The radiolabelled Ga complex was prepared to a high radiochemical purity (> 97%, HPLC) followed by initial biodistribution data with the significant tumour accumulation of the tracer at two hours, which is far higher than free Ga-67 cation, while the compound wash-out is significantly faster. CONCLUSION: The above-mentioned pharmacokinetic properties suggest an interesting radiogallium complex prepared by the PET Ga radioisotope, (68)Ga, in accordance with the physical half life, for use in fibrosarcoma tumours and possibly in other malignancies.

Preparation and evaluation of [(61)Cu]-thiophene-2-aldehyde thiosemicarbazone for PET studies.

BACKGROUND: [(61)Cu]Thiophene-2-aldehyde thiosemicarbazone ([(61)Cu]TATS) (4) was prepared according to an analogy of carrier copper compound with antitumor activity, for eventual use in PET. MATERIAL AND METHODS: [(61)Cu]TATS was prepared using copper-61 acetate and in-house made ligand (TATS) in one step. (61)Cu was produced via the (nat)Zn(p,x) (61)Cu nuclear reaction (180 mircoA, 22 MeV, 3.2 h) followed by a two-step chromatography method (222 GBq of (61)Cu(2+)). [(61)Cu]TATS preparation was optimized for reaction conditions (buffer concentration and temperature). The tracer was finally administered to normal rats for biodistribution studies. RESULTS: Total radiolabelling of the tracer took 30 minutes with a radiochemical purity of more than 90% (using HPLC and RTLC) and specific activity of about 250-300 Ci/mmol. The complex was stable in the presence of human serum for an hour. The biodistribution of copper cation and the tracer was checked in wild-type rats for up to 2 hours with significant spleen and lung uptake of the tracer. CONCLUSIONS: The production of (61)Cu via the (nat)Zn(p,x) (61)Cu is an efficient and reproducible method with high specific activity leading to the production and preliminary evaluation of [(61,)Cu]TATS, a potential PET tracer, was reported.

Porphyrins as ligands for 64copper: background and trends.

Porphyrins and 64Cu have emerged as a novel synergic option for applications in PET molecular imaging. Both the characteristics and photophysical properties of macrocyclic porphyrins and the relatively long half-life of the copper isotope, in addition to the increased tumor-specific uptake of porphyrins compared to normal cells, make this complex an attractive option not only for diagnosis but also for therapeutic applications. Herein, we present an overview of the latest results on the development of PET agents based on porphyrins and 64Cu, including methods used to improve the selectivity of these macrocycles when conjugated with biological units such as monoclonal antibodies, peptides or proteins.

Production and biological evaluation of [18F]-6-thia-14-fluoro-heptadecanoic acid.

BACKGROUND: [(18)F]-6-thia-14-fluoro-heptadecanoic acid 3b, a free fatty acid, has been used in myocardial PET imaging. In order to establish an automated synthesis module for routine production in the country, a study was performed for optimization of the production conditions as well as making modifications. MATERIAL AND METHODS: [(18)F]Benzyl-14-Fluoro-6-thia-heptadecanoate 2b was prepared in no-carrier-added (n.c.a) form from Benzyl-14-tosyloxy-6-thia-heptadecanoate 1 in one step at 90 degrees C in Kryptofix2.2.2/[(18)F] with acetonitrile as the solvent followed by Silica column chromatography. The radiolabelled ester 2 was then hydrolysed to yield [(18)F]-6-thia-14-fluoro-heptadecanoic 3b. The final solution was concentrated using the C(18) SPE system and administered to normal rats for biodistribution and co-incidence imaging studies. RESULTS: The synthesis took 15 min with overall radiochemical yield of 15-25% (EOS) and chemical-radiochemical purity of more than 90%. Automation was performed using a two-pot synthesis. The best imaging time was shown to be 140-180 minutes post injection. CONCLUSIONS: Using this procedure a fast, reliable, automated synthesis for the cardial PET tracer, i.e. [(18)F]FTHA, can be obtained without an HPLC purification step.

Grafting of [(64)Cu]-TPPF20 porphyrin complex on Functionalized nano-porous MCM-41 silica as a potential cancer imaging agent.

Mesoporous silica, MCM-41, functionalized with 3-aminopropyltriethoxysilane (APTES) was investigated as a potential drug delivery system, using [(64)Cu]-5, 10, 15, 20-tetrakis penta fluorophenyl porphyrin complex. [(64)Cu]-TPPF20 complex was grafted on functionalized MCM-41. The product was characterized by paper chromatography, FTIR spectroscopy, low angle X-ray diffraction, CHN and TGA/DTA analyses and atomic force microscopy. The biological evaluations of the grafted complex, [(64)Cu]-TPPF20@NH2-MCM-41, were done in Fibrosarcoma tumor-bearing Sprague-Dawley rats using scarification studies and Sopha DST-XL Dual-Head SPECT system. The actual loading amount of aminopropyl groups was found about 1.6mmol per gram of final silica. The specific activity of the final compound was found to be 3Ci/g. Amine functionalized MCM-41 was found to be a good platform for theranostic radiopharmaceuticals such as copper-64 complexes. Considering the accumulation of the tracer in tumor cells, fast wash-out from normal tissues, the short half-life copper-64 and less imposed radiation doses to patients, [(64)Cu]-TPPF20@NH2-MCM-41 can potentially be a suitable candidate for tumor imaging applications and future PET studies.

Production of 68Ga-citrate Based on a SnO2 Generator for Short-Term Turpentine Oil-Induced Inflammation Imaging in Rats.

INTRODUCTION: Gallium-68 citrate has been successfully applied in the PET imaging of infections and inflammation in some centers; however further evaluation of the tracer in inflammation models is of great importance. METHODS: 68Ga-citrate prepared from [68Ga]GaCl3 (eluted form an SnO2 based 68Ge/68Ga generator) and sodium citrate at optimized conditions followed by quality control tests was injected to normal and turpentine-oil induced rats PET/CT imaging studies up to 290 min. RESULTS: 68Ga-citrate was prepared with acceptable radiochemical purity (>99 ITLC, >99% HPLC), specific activity (28-30 GBq/mM), chemical purity (Sn, Fe <0.3 ppm; Zn<0.2 ppm) in 15 min at 50°C. PET/CT imaging of the tracer demonstrated early detection of inflamed site in animal models in 60-80 min. CONCLUSION: This study demonstrated possible early detection of inflammation foci in vivo using 68Ga-citrate prepared using commercially available 68Ge/68Ga generators for PET imaging.

Development of a (68)Ga-peptide tracer for PET GnRH1-imaging.

OBJECTIVE: Total synthesis, quality control and preclinical evaluation of [(68)Ga]-DOTA-triptorelin ([(68)Ga]-DOTA-TRP) is reported as a possible PET radiotracer for GnRH receptor imaging. METHODS: DOTA-TRP was totally synthesized in two steps and after characterization went through radiolabelling optimization studies followed by tracer stability. The biodistribution of the tracer in normal male rats and 4T1 tumour-bearing mice was performed in 120 min after i.v. injection. RESULTS: The peptide and the conjugates were synthesized with >95 % chemical purity. [(68)Ga]-DOTA-TRP complex was prepared in high radiochemical purity (>99 %, ITLC, HPLC) and specific activity of 1400-2100 MBq/nM at 95 °C using 40-60 µg of the peptide in 5-7 min followed by solid phase purification. The IC50 [nM] DOTA-TRP was comparable to the intact peptide, 0.11 ± 0.01 and 0.22 ± 0.05, respectively. The biodistribution of the tracer demonstrated kidney, stomach, and testes significant uptake, all in accordance with GnRH receptor ligands. Significant tumour uptake was observed in 4T1 tumour-bearing female mice 30-120 min post-injection with tumour:blood and tumour:muscle ratios of 28 and >50 in 60 min, respectively. Kidney is rapidly washed from the tracer. [(68)Ga]-DOTA-TRP can be proposed as a possible tracer for GnRH-R imaging studies.

Current Status of Nuclear Medicine Practice in the Middle East.

The practice of nuclear medicine (NM) in the Middle East region has experienced an important growth in the last 2 decades and has become crucial in providing healthcare to the region's population of about 395 million people. Even though there are some countries in which the services provided are limited to basic coverage of studies with (99m)Tc and (131)I, most have well-established practices covering most of the available studies in this medical specialty; this is the case in for example, Iran, Israel, Kuwait, Saudi Arabia, and Turkey. According to data provided by the NM professionals in the 17 countries included in the present publication, which was collected by the International Atomic Energy Agency in 2015, the total number of gamma cameras in the region is 910 with an average of 2.3 gamma cameras per million inhabitants. Out of these, 107 cameras, or 12%, are SPECT/CT cameras. There are 194 operating PET/CT scanners, translating to one PET/CT scanner for 2.04 million people on average. The availability of PET/CT scanners in relation to population is the highest in Lebanon and Kuwait, with 2.2 and 1.7 scanners per million people, respectively. There is a total of 628 NM centers in the 17 countries, whereas most NM centers belong to the public healthcare system and in most of the countries are widely spread and not confined exclusively to capital cities. As for the radionuclide therapies, (131)I is used regularly in diagnostic workup as well as in therapeutic applications in all the countries included in this analysis. Only five countries have the capability of assembling (99)Mo-(99m)Tc generators (Egypt, Iran, Saudi Arabia, Israel, and Turkey), and cold kits are produced in several countries. Although there are no capabilities in the region to produce (99)Mo from nuclear reactors, a total of 46 cyclotrons are operated for production of PET radionuclides. The most widely used PET tracer in the region is (18)F-FDG followed by (18)F-NaF; concomitantly, the availability of (68)Ge-(68)Ga generators is increasing and studies involving prostate-specific membrane antigen or DOTA-chelated peptides or both are performed in at least seven countries. Although therapeutic radionuclide agents are mostly imported from outside the region, this does not limit the availability of therapies with (90)Y, (153)Sm, (177)Lu, (131)I, (188)Re, and (89)Sr. Nevertheless, therapies based on alpha particle emitters are still largely not available in the region and are currently only available in Israel and Turkey. Regarding human resources, according to the data provided there are 1157 NM physicians, 1953 technologists, 586 medical physicists, and 173 radiopharmacists or radiochemists in the region. Approximately half of all available human resources are accounted for by Turkey. The region has great potential for expanding the applications of NM; this becomes especially important in view of the high prevalence of non-communicable diseases. Further increasing awareness of the clinical applications of NM in healthcare and strengthening technical and human capacities including the establishment of training programs for all professionals and disciplines in the field are recognized as key components in advancing the practice of NM in the Middle East.