An improved method for the preparation of [<Superscript>14</Superscript>C]thiourea of high radiochemical purity

Summary An improved method for the preparation of [¹⁴C]thiourea of high radiochemical purity is described. [¹⁴C]thiourea is prepared by the barium cyanamide route and is purified by vacuum-sublimation. The labeled product showed ammonium [¹⁴C]thiocyanate as a radiochemical impurity in the range of 2-4%. This was further purified by silica-gel column chromatography to get the product having more than 99% radiochemical purity.     

Synthesis of ammonium [14C]thiocyanate

Ammonium [¹⁴C]thiocyanate was prepared from potassium [¹⁴C]cyanide with a radiochemical yield of 90%. [¹⁴C]hydrocyanic acid, generated from potassium [¹⁴C]cyanide by sulphuric acid, reacts with aqueous ammonia and elemental sulphur in the presence of trace amounts of ammonium sulphide to yield ammonium [¹⁴C]thiocyanate.     

Source tracing of chromium-, manganese-, nickel- and zinc-containing particles (PM<Subscript>10</Subscript>) by micro-PIXE spectrum

Due to the interesting anti-proliferative properties of gallium-thiosemicarbazone complexes, the production of [⁶⁷Ga]labeled 2-acetylpyridine 4,4-dimethyl thiosemicarbazone (APTSM₂) was investigated. The freshly prepared [⁶⁷Ga]GaCl₃ was mixed with 2-acetylpyridine 4,4-dimethyl thiosemicarbazone for 60 minutes at 90 °C to yield [⁶⁷Ga]APTSM₂ with a radiochemical yield of more than 98%. Radio-thin-layer-chromatography (RTLC) showed a radiochemical purity of more than 99%. A specific activity of about 370–740 MBq/mmol (10–20 Ci/mmol) was obtained. The stability of the final product was checked in the absence and presence of human serum at 37 °C. The partition coefficient of the final complex was also determined. The biodistribution of the labeled compound in normal rats was compared with that of free Ga³⁺ cation up to 22 hours.     

Facile conversion of [1-<Superscript>14</Superscript>C]lauronitrile to [1-<Superscript>14</Superscript>C]lauric acid under microwave irradiation

Summary A highly efficient and an optimized synthesis of [1-¹⁴C]lauric acid with high specific activity (50 mCi/mmol) is described. [1-¹⁴C]lauric acid was prepared from [1-¹⁴C]lauronitrile, in 2 minutes with a mixture of concentrated hydrochloric acid: propionic acid (1: 2 v/v) under microwave irradiation, in quantitative yield.     

Preparation and preliminary evaluation of [<Superscript>67</Superscript>Ga]-tetra phenyl porphyrin complexes as possible imaging agents

[⁶⁷Ga]labeled tetraphenyl porphyrin ([⁶⁷Ga]-TPP) was prepared using freshly prepared [⁶⁷Ga]GaCl₃ and tetraphenyl porphyrin (TPPH₂) for 30–60 min at 25 °C (radiochemical purity: >97 ± 1% ITLC, >98 ± 0.5% HPLC, specific activity: 13–14 GBq/mmol). Stability of the complex was checked in final formulation and human serum for 24 h. The partition coefficient was calculated for the compound (log P 1.89). The biodistribution of the labeled compound in vital organs of wild-type rats was studied using scarification studies and SPECT imaging up to 24 h. A detailed comparative pharmacokinetic study performed for ⁶⁷Ga cation and [⁶⁷Ga]-TPP. The complex is mostly washed out from the circulation through kidneys and can be an interesting tumor imaging/targeting agent due to low liver uptake and rapid excretion through the urinary tract.     

Development of [<Superscript>64</Superscript>Cu]-DOTA-anti-CD20 for targeted therapy

Copper-64 was produced as a by-product of ⁵⁵Co via ⁶⁴Ni(p,n)⁶⁴Cu by 15 MeV proton bombardment of ^natNi resulting in a thick target yield of 5.31 MBq/μAh (143.5 μCi/μAh) and a radiochemical separation yield of 95% (radionuclide purity >97% after 25 hours of bombardment). Rituximab was successively labeled with [⁶⁴Cu]-CuCl₂. N-succinimidyl-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA-NHS) was prepared at 25 °C using DOTA and N-hydroxy succinimide (NHS) in CH₂Cl₂ followed by the addition of 1 ml of a Rituximab pharmaceutical solution. Radiolabeling was performed at 37 °C in 3 hours. Radio thin-layer chromatography showed an overall radiochemical purity of 90–95% at optimized conditions (specific activity=30 GBq/mg, labeling efficacy; 82%) using various chromatography systems. The final isotonic ⁶⁴Cu-DOTA-Rituximab complex was passed through a 0.22 μm filter and checked by gel electrophoresis for radiolysis control. Stability of the final product was checked in the formulation and in presence of human serum at 37 °C.     

Preparation and evaluation of [<Superscript>201</Superscript>Tl](III)-DTPA complex for cell labeling

Due to interesting physical properties and wide availability of ²⁰¹Tl as a SPECT radionuclide, the incorporation of this nuclide into DTPA for cell labeling was targeted. Thallium-201 (T _1/2 = 3.04 d) in Tl⁺ form was converted to Tl³⁺ cation in the presence of O₃/6M HCl and di-isopropyl ether, controlled by RTLC/gel electrophoresis methods. The final evaporated activity reacted with cDTPA in normal saline to yield [²⁰¹Tl](III)DTPA at room temperature after 0.5 hour, followed by solid phase extraction purification using C₁₈ Sep-Pak column (radiochemical yield >95%). Radiochemical purity of more than 99% was obtained using RTLC with specific activity of about 260 GBq/mmol. The stability of the tracer was checked in the final product in the presence of human serum at 37 °C up to 3 days. The partition coefficient was also measured. The labeled compound was used in red blood cell (RBC) labeling. The cell uptake ratio was determined at 4, 25 and 37 °C up to 3 hours.     

Preparation of <Superscript>125</Superscript>I-celecoxib with high purity as a possible tumor agent

A preparation of ¹²⁵I-celecoxib is carried out via an electrophilic substitution reaction. The reaction parameters studied were celecoxib concentration, reaction temperature, pH of the reaction mixture and kinds of oxidizing agents in order to obtain a high radiochemical yield of the ¹²⁵I-celecoxib. Using 3.7 MBq of Na¹²⁵I, 150 μg (3.9 mM (mmol/L)) of celecoxib, and 1.6 mM (mmol/L) of chloramine-T (CAT) as oxidant at pH 4 and 60 °C for 15 min a maximum radiochemical yield of ¹²⁵I-celecoxib (65%) was obtained. The labeled compound was separated and purified by means of high pressure liquid chromatography (HPLC). The biological distribution in infected mice indicates the suitability of radioiodinated celecoxib as imaging of tumor.     

Facile syntheses of isotope-labeled chiral octahydroindole-2-carboxylic acid and its <Emphasis Type="Italic">N</Emphasis>-methyl analog

We have synthesized deuterium and carbon-14 labeled enantiomerically pure octahydroindole-2-carboxylic acid (PD0140417), N-methyl octahydroindole-2-carboxylic acid (PD0348183) and their racemic analogs (PD0108405 and PD0338055). [ring-U-¹⁴C]PD0140417 was prepared from [ring-U-¹⁴C]benzoic acid in a seven-step synthesis in 6.2% overall radiochemical yield. [¹⁴C]PD0348183 was prepared from [¹⁴C]BaCO₃ in a five-step synthesis in 16% radiochemical yield. Additionally, [D]PD0108405 and [D]PD0338055 were synthesized by direct platinum-catalyzed hydrogenation with deuterium gas.     

Synthesis of <Emphasis Type="Italic">O</Emphasis>-[2-[<Superscript>18</Superscript>F]fluoro-3-(2-nitro-1<Emphasis Type="Italic">H</Emphasis>-imidazole-1-yl)propyl]tyrosine ([<Superscript>18</Superscript>F]FNT]) as a new class of tracer for imaging hypoxia

For detection of hypoxic tumor tissue, all radiotracers synthesized until now, are based on the concept that cellular uptake is being controlled by diffusion. As a new approach, we chose the concept to have the tracer hypothetically transported into the cells by well known carrier systems like the amino acid transporters. For this purpose, radiosynthesis of O-[2-[¹⁸F]fluoro-3-(2-nitro-1H-imidazole-1yl)propyl]tyrosine ([¹⁸F]FNT]) was carried out from methyl 2-(benzyloxycarbonyl)-3-(4-3-(2-nitro-1H-imidazol-1-yl)-2-(tosyloxy)propoxy) phenyl)propanoate via no-carrier-added nucleophilic aliphatic substitution. After labelling, 81 ± 0.9% of labelled intermediate i.e. methyl 2-(benzyloxycarbonyl)-3-(4-(2-[¹⁸F]fluoro-3-(2-nitro-1H-imidazole-1-yl)propoxy) phenyl)propanoate was obtained at 140 °C. At the end of radiosynthesis, [¹⁸F]FNT was obtained in an overall radiochemical yield of 40 ± 0.9% (not decay corrected) within 90 min in a radiochemical purity of >98% in a formulation ready for application in the clinical studies for PET imaging of hypoxia.     

One-step conjugation of glycylglycine with [<Superscript>18</Superscript>F]FDG and a pilot PET imaging study

This study describes a single step conjugation of Glycylglycine (GlyGly) which is a small peptide, with [¹⁸F]FDG via oxime formation. Amiooxy-functionalization of GlyGly (AO-GlyGly) was accomplished through the reaction of Boc-aminooxy succinimide ester. Conjugation reaction was performed at 100 °C for 30 min in a vial containing AO-GlyGly and [¹⁸F]FDG solution. The radiolabeled product ([¹⁸F]FDG-GlyGly) was obtained with 98.65?±?0.35% yield without any purification step which makes this method more attractive for ¹⁸F radiolabeling. The present study is concluded with an in vivo pilot animal PET study to assess biodistribution and kinetics of chemoselectively [¹⁸F]FDG tagged GlyGly in vivo.     

A convenient method for <Superscript>14</Superscript>C-labeling of <Emphasis Type="Italic">N</Emphasis>-(2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)-1H-pyrrole-2-carboxamide-[carboxy-<Superscript>14</Superscript>C] as CCK-A antagonist

N-(2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)-1H-pyrrole-2-carboxamide-[carboxy-¹⁴C] was prepared as part of a five-step sequence from pyrrol-2-carbonitrile-[cyano-¹⁴C] as a key synthetic intermediate which has been synthesized from 2-bromopyrrole and zinc [¹⁴C]-cyanide in the presence of tetrakis (triphenylphosphine)palladium.     

Synthesis and analysis of [4−14C] phenylbutazone

[4?¹⁴C] phenylbutazone (4-butyl-1,2-diphenyl-3,5-[4?¹⁴C] pyrazolidinedione) has been synthesised on the 5 mmole scale. [2?¹⁴C] Diethyl malonate was reacted with n-butyl bromide and the product condensed with hydrazobenzene in the same reaction vessel. The radiochemical yield of highly purified product was 48%. It is proposed that care should be taken in analyses for metabolic products, particularly by TLC, since 4-hydroxyphenylbutazone was shown to be an oxidation product of phenylbutazone in solution and on TLC plates under quite mild conditions. A pure sample of 4-hydroxy [4?¹⁴C] phenylbutazone was isolated.     

Development of <Superscript>111</Superscript>In labeled insulin for receptor imaging/therapy

In order to target insulin receptors in various diabetic and insulinoma conditions, human recombinant insulin was successively labeled with [¹¹¹In]-indium chloride after conjugation with freshly prepared cyclic DTPA-dianhydride (ccDTPA). The best results of the conjugation were obtained by addition of 0.5 mL of an insulin pharmaceutical solution (5 mg/mL, in phosphate buffer, pH 8) to a glass tube pre-coated with DTPA-dianhydride (0.01 mg) at 25 °C with continuous mild stirring for 30 minutes. Radio-thin layer chromatography (RTLC), instant thin layer chromatography (ITLC) and high performance liquid chromatography (HPLC) have shown an overall radiochemical purity of higher than 93% at optimized conditions (specific activity = 550–750 MBq/mg, radiochemical yield =81%). The white blood cell labeling capacity of the tracer was determined up to 4 hours at 37 °C. Preliminary in vivo studies in normal rat model was performed to determine the biodistribution of the radiotracer up to 48 hours. It showed a high liver and spleen uptake of the tracer which is consistent with other reported radiolabeled insulins. SPECT images have also shown high liver accumulation of the tracer.     

Synthesis of <Emphasis Type="Italic">O</Emphasis>-(2-[<Superscript>18</Superscript>F]fluoroethyl)-<Emphasis Type="Italic">L</Emphasis>-tyrosine and its biological evaluation in B16 melanoma-bearing mice as PET tracer for tumor imaging

O-(2-[¹⁸F]fluoroethyl)-L-tyrosine ([¹⁸F]FET), a fluorine-18 labeled analogue of tyrosine, has been synthesized and biologically evaluated in tumor-bearing mice. The whole synthesis procedure is completed within 50 min. The radiochemical yield is about 40% (no decay corrected) and radiochemical purity more than 97% after simplified solid phase extraction. [¹⁸F]FET shows rapid, high uptake and long retention in the tumor as well as low uptake in the brain. The ratios of tumor-to-muscle (T/M) and tumor-to-blood (T/B) of [¹⁸F]FET are similar to those of [¹⁸F]FDG, but the ratios of tumor-to-brain (T/Br) are 2–3 times higher than that of [¹⁸F]FDG. Autoradiography of [¹⁸F]FET demonstrates a remarkable accumulation in melanoma with high contrast. It appears to be a probable competitive candidate for melanoma imaging with PET. Supported by the Knowledge Innovation Project of Chinese Academy of Sciences (No. KJCX1-SW-08) and the National Natural Science Foundation of China (Grant No. 30371634)     

Synthesis of radioiodinated 4-[*I]iodoantipyrine via isotopic exchange

Radioiodinated 4-[*I]iodoantipyrine labeled with radioiodine (i.e., ¹²³I or ¹²⁵I or ¹³¹I) has been used for modeling radiation damage on cell nuclei of tumor cells where the characteristic high linear energy transfer (high-LET) of the Auger electron could be demonstrated. Also, the compound is currently used for the measurement of regional cerebral blood flow (rCBF) in autoradiography. 4-[¹³¹I]iodoantipyrine was synthesized by two methods via a nucleophilic isotopic exchange reaction between ¹³¹I as iodide ion [¹³¹I^–] and inactive 4-[¹²⁷I]iodoantipyrine: either in absolute ethyl alcohol catalyzed by ammonium acetate or in dry state molten ammonium acetate (m.p. 114 °C) as an isotopic exchange medium without carrier addition. The first one is called wet method: where a solution of 4-iodoantipyrine and ammonium acetate in absolute ethyl alcohol and lyophilized Na¹³¹I was heated briefly up to boiling (80 to 90 °C) for 30 minutes under reflux. The second one is called dry state-molten method: where the alcoholic solution containing 4-iodoantipyrine and ammonium acetate and the lyophilized Na¹³¹I were heated briefly in a nitrogen stream to dryness at 120 to 125 °C for 5 minutes or melted by gradual heating at 150 to 160 °C for 5 minutes. A radiochemical yield ranged between 90%–95% in each method has been obtained for 4-[¹³¹I]iodoantipyrine. In both methods, the reaction proceeds properly without carrier addition by an addition – elimination mechanism. The physico-chemical parameters affecting the radiochemical yield of the isotopic exchange reaction [i.e., reaction time, temperature, exchange medium, concentration of the reactants, carrier (KI) addition and pH] were investigated. Chromatographic analysis i.e., TLC and HPLC were used to determine the radiochemical yield as well as the purity of the final product, which was as pure as 99.9%.     

Production and quality control of [<Superscript>166</Superscript>Ho]-DOTA-bevacizumab for therapeutic applications

Antiangiogenic monoclonal antibodies in combination with therapeutic radionuclides are potential targeted therapy agents in cancer. In this study, bevacizumab was successively labeled with [¹⁶⁶Ho]HoCl₃ after conjugation with DOTA-NHS-ester with a radiochemical purity of higher than 95% (RTLC). The conjugates were purified by molecular filtration, the average number of DOTA conjugated per mAb was calculated and total concentration was determined by spectrophotometric method and the average chelate to antibody ratio (c/a) for the conjugate used in this study was 5.8:1 and protein integrity experiments (SDS-PAGE). The biodistribution studies in wild-type rats demonstrate a similar pattern to the other radiolabeled anti-vascular endothelial growth factor A (VEGF-A) immunoconjugates. ¹⁶⁶Ho-DOTA-bevacizumab is a potential compound for therapy/imaging of VEGF-A expression in oncology.     

Production of carrier free <Superscript>188</Superscript>Re radioisotope generator based on aluminum tungstate matrix

Improved radionuclide generator include a substantially insoluble salt of a radioactive parent which may be directly packed in column for subsequent elution of the daughter radionuclide. An improved ¹⁸⁸Re generator was prepared by reacting a radioactive tungsten (¹⁸⁸W) as parent radionuclide incorporated with aluminum chloride to obtain an insoluble radioactive aluminum tungstate matrix. The investigated matrix was characterized on the basis of the chemical composition, IR, thermal analysis and mechanical stabilities. The factors affecting the elution performance were studied such as influence of pH, molar ratio and drying temperature. From the obtained data, the molar ratio W:Al was 1.5:1 at pH = 4, the matrix dried at 105 °C for 2 h. Chromatographic and multichannel analysis has been currently used to investigate the radiochemical and radionuclidic purity respectively on eluted ¹⁸⁸Re. An elution yield more than 80%, with radiochemical purity <98% and radionuclidic purity <99% with a ¹⁸⁸W break through >10⁻⁴% of the column. The Al⁺³ and W contents value were about 2 and 3 μg/mL eluate. The obtained data approved the stability of the prepared generator and its suitability for medical application.     

Synthesis,bioevaluation and gamma scintigraphy of <Superscript>99m</Superscript>Tc-N-2-(furylmethyl iminodiacetic acid) complex as a new renal radiopharmaceutical

A ligand of N-2-(furylmethyl iminodiacetic acid) (FMIDA) has been easily labeled by a tetradentate chelating agent of [^99mTc]. Factors like a stannous chloride solution as a reducing agent (100 μg), substrate amount (100 μg), pH (7), in vitro stability (8 h) and temperature (37 °C) have been systematically studied to optimize high radiochemical yield (98.0%). The radiochemical conversion was calculated on thin-layer chromatography, paper electrophoresis, and high performance liquid chromatography. Biodistribution study showed that this complex was removed from the kidneys and bladder path way during 1 h post injection. Therefore, [^99mTc]FMIDA may be used as renal function radiotracer.     

Development of an in vivo radionuclide generator by labeling bleomycin with <Superscript>191</Superscript>Os

Bleomycin (BLM) has been labeled with various radioisotopes and widely used in therapy and diagnosis. ¹⁹¹Os is a parent radionuclide with 15.4 day half-life and decays by beta emission to ^191mIr, which is a radionuclide with 4.96 s half-life. BLM was labeled with ¹⁹¹Os-hexachloro-osmate and its distribution and stability in wild-type mice was determined. The complex was obtained at the pH 2 in normal saline at 90 °C in 48 h. Radio-TLC showed an overall radiochemical yield of 93–97%, radio-chemical purity > 97%. The biodistribution study for ¹⁹¹Os-hexachloro-osmate and ¹⁹¹Os-BLM were carried in wild type-mice up to 14 days. Lungs, liver and spleen uptake increased 24–72 h after administration of ¹⁹¹Os-BLM. 24 h after administration, the radioactivity of the kidney increased and remained constant.