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Rold1, 4, Silvia S. Jurisson5, Charles J. Results The metallated conjugates were produced in high radiochemical yield and displayed very high binding affinity for GRPR 8.
Low accumulation and rapid clearance of tracer in normal tissues was observed. Conclusions Favorable pharmacokinetics for the 90Y-radiolabeled tracer in in vivo studies support further investigations for microPET imaging or therapy in animal models. Thus, radiolabeled with a positron emitter this strategy represents a highly attractive platform for the design of radiopharmaceuticals for PET imaging of various oncological entities and for therapeutic applications.
Methods Conjugation with tris- bromomethyl benzene via the three reactive cysteines of a partly randomized amino acid sequence and fusion to the phage geneprotein generated a library of bicyclic peptide conjugates with two peptide loops . A subsequent affinity selection process yielded several binders with variants showing single digit nanomolar affinity and high specificity for MMP The selected optimal variant was conjugated to DOTA and analyzed preclinically according to its cell binding and in vivo tumor-targeting properties.
Organ distribution at 30 min. Conclusions The phage display selection of bicyclic peptides was shown to be a highly flexible and attractive strategy for generating and developing peptidic drug leads.
With the Bicyclic DOTA-compounds being roughly Da in size, organ distribution studies revealed MMPselective tumor uptake in the xenograft model and rapid clearance from non-target crucial organs resulting in high imaging contrasts.
Directed against tumor specific membrane-type biological targets such as MMP and labeled with radiometals these molecules represent highly promising radiopharmaceuticals for future clinical PET imaging and radioendotherapy of various oncological diseases.
Louis, Missouri, United States Objectives Radiopharmaceuticals capable of imaging the functional transport activity of breast cancer resistance protein BCRP, ABCG2 are desired to assist in the stratification of chemotherapeutic choices. Galmydar, a fluorescent and moderately hydrophobic Ga III monocationic complex, was evaluated as a substrate capable of monitoring BCRP functional transport activity.
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To assess the sensitivity and specificity of Galmydar to probe BCRP-mediated efflux activity in cellulo, cellular accumulation was determined in transfected cells. Methods For cellular accumulation studies, live-cell fluorescence microscopy and 67Ga-radiotracer uptake assays were performed.
Results Cellular accumulation profiles of Galymdar in HEK cells, measured via live-cell fluorescence microscopy and 67Ga-radiotracer uptake assays, are inversely proportional to BCRP expression. Conclusions Live-animal imaging data coupled with cellular accumulation data establishes Galmydar as a template scaffold for development of a PET tracer for imaging BCRP-mediated functional transport in vivo.
Acknowledgements We thank Julie L. Prior for technical assistance.
An imaging agent assessing Met status would aid in diagnosis, patient selection for Met-targeted therapies and monitor therapeutic responses. Previously, we described a peptide targeting Met labeled with [99mTc] AH which exhibited nM affinity and targeting to warrant further development for human use .
Conclusions [94mTc]MetP bound to Met with high affinity and had uptakes correlating with Met expression levels in vitro and in vivo.
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These results suggest that [94mTc]MetP may have potential to identify patients whose tumors express moderate-high levels of Met in tumors and therefore, who may benefit from Met-targeted therapies. Recent publications showed that Tenascin C expression by cancer lesions predicts tumor growth, metastasis, and angiogenesis, suggesting Tenascin C as a potential therapeutic target. Currently there is no noninvasive method to determine tumoral Tenascin C expression in vivo. Methods Tenascin C aptamer was radiolabeled with 18F and 64Cu was found to be stable in vitro and in vivo.
PET imaging studies for the evaluation of tumor uptake and pharmacokinetics of Tenascin C aptamer were performed in comparison to a non-specific scrambled aptamer Sc aptamer. The uptake of Tenascin C aptamer was significantly higher than that of Sc aptamer in Tenascin Cpositive tumors. The labeled Tenascin C aptamer had fast clearance from the blood and other nonspecific organs through the kidneys, resulting in high tumor contrast.
Conclusions Our data suggest that suitably labeled Tenascin C aptamer can be used as a PET tracer to image tumor expression of Tenascin C with a high tumor-to-background ratio and might provide insightful and personalized medical data that will help determine appropriate treatment and monitoring.
Vugts3, Guus A. To date, 89Zr -based imaging probes are obtained exclusively through derivatives of desferrioxamine DFO , a chelator which does not complete the octadentate coordination sphere of the radiometal. There is compelling preclinical evidence that the incomplete coordination of 89Zr by DFO is responsible for the observed instability of the chelate in vivo.
The release of the radiometal from the tracer results in the non-specific accumulation of the radiometal in radiation sensitive bones, a potential safety issue for clinical applications. Methods Herein, we wish to report the rational design, DFT-calculation guided synthesis, and in vitro evaluation of the first octadentate bifunctional chelating agent BFCA for the development 89Zr-based radiopharmaceuticals with an improved stability profile.
References  Fischer, G. Molecules , 18,  Patra M.
Stability, specificity, affinity, cellular processing, and pharmacokinetics were studied. Results The conjugates were synthesized, stably labeled with 68Ga and In with retained binding specificity.
IC50 were in low nanomolar range. In mice bearing PC-3 tumors conjugates demonstrated specific uptake in tumors. The influence of chelators on biodistribution of conjugates was essential and differed between 68Ga and In. Stability in human serum was checked after labeling with 68Ga.
T2weighted MR 3. After i. The presence of IO in tissue was. Existence of ferric iron was confirmed in the extracted liver and spleen. Figure 1. The design of peptidomimetics to mimic these structural elements is essential to effectively target these interactions in the design of diagnostics and therapeutics.
The objective of this project is to design and synthesize a novel amino acid containing bipyridine to allow for the incorporation of a bipyridine chelator into a peptide by standard solid-phase methods. Methods The bipyridine amino acid was synthesized by Ullmann coupling of 3-nitrochloropyridine, followed by reduction to the diamine, addition of a succinic group to one of the amines, followed by protection of the free amine with an Fmoc group.
Characterization by CD spectroscopy suggests the retention of the secondary structure upon complexation to Re.
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HPLC analysis showed a peak with a similar retention time as the Re standard. References  Whitby, LR et al. The objective of the present work is to prepare a new zinc bioconjugate derivative of folic acid 1 that can be used to prepare new technetium and rhenium radiopharmaceuticals with this biomolecule following this new approach.
Next, the reaction between this compound and trifluoroacetic acid  yielded to compound 2. Finally, the zinc folate derivative 4 was prepared by means of the reaction between the succinimidyl ester of the zinc II dithiocarbamate of isonipecotic acid 3 and compound 2 using previously reported methods .
Results The activated ester group of the zinc complex 3 reacts with the amine group of the HMDA-folic acid compound 2 , which was synthesised from folic acid 1 using previously reported methods [2, 3], to yield the target metal complex 4. Conclusions We have successfully synthesised a new bioconjugated zinc complex of folic acid useful to prepare radiopharmaceuticals with high specific activity.
References  Lecina, J. These ligands rapidly form highly stable CuII complexes that are resistant to metal leaching. The ligand structure enables convenient introduction of additional conjugatable groups like -COOH groups to allow coupling to various biomolecules.
Two coupling groups such as maleimide 1 or isothiocyanate 2 have been attached for further conjugation. These ligands were then conjugated to the thiolated dPGS via Michael addition of 1 and also via direct labeling of 2 to dPGS amine to yield highly stable conjugates.
Challenge experiments were conducted in presence of EDTA or copper seeking superoxide dismutase SOD for evaluation of in vitro stability. Biodistribution and PET studies were conducted in male Wistar rats. Conclusions DMPTACN ligands are a versatile platform which can be applied for labeling small molecules, proteins as well as polymers for detailed insight on their biodistribution profiles using 64Cu that allows studying relatively long biochemical processes and thus, prove to be an attractive candidate for PET imaging.
Goscinski1, Timothy A. Wencewicz1, Suzanne E. Lapi2 1 Chemistry, Washington University in St. Zirconium is an emerging radiometal with chemistry similar to iron. Therefore, we are investigating 89Zr-siderophores as a promising class of imaging agents for assessment of bacterial infection. Cells were labeled in LB Miller broth, human serum and 0. Labeled cultures were incubated for various amounts of time, the cells isolated and cell-associated activity quantified.
Additionally, the Gram-positive and -negative bacteria exhibited different uptake behavior: S. The media in which cells were grown and labeled greatly affected 89Zr-DFO uptake behavior, as did length of incubation and iron availability during growth.
Experiments examining the dose- and time-dependence of the uptake are ongoing. Conclusions We have shown that 89Zr-DFO is rapidly taken up by bacteria and that the extent of this uptake and internalization depends on the type of bacteria exposed to the complex as well as the conditions under which the exposure occurs.
Currently, our group and others  are working on new urea-based PSMA ligands suitable for endoradiotherapy of prostate cancer PCa. Their in vivo tumor-targeting properties have been optimized by modification of the linker regions.
By an effort to elucidate the structure-activity relationships SAR of up to 30 new PSMA ligands, the tailor-made candidate PSMA has been identified with the putatively best in vivo characteristics. Organ distribution revealed specific uptake in LNCaP tumors 8. PSMA exhibited a rapid clearance from the kidneys to 2.
Conclusions The overall characteristics of the here presented new PSMA ligands could be realized successfully by systematic chemical modification of the respective linker region.
Tailor-made PSMA exhibits high PSMAspecific tumor uptake, rapid background and fast kidney excretion and is, thus, ready for first-in-man theranostic studies. However, the radiation dose to the kidneys has been a concern for the possibility of repeated imaging studies in humans.
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The organ distribution data in each species was extrapolated to those of a human, assuming similar distribution pattern. Results The extrapolated human whole body effective dose was 0. The absorbed dose to the kidneys was the limiting parameter: 0.
Thomas N. Bulenga was supported by the Swedish Institute Scholarship. It affects functionality of vital organs such as liver, lung, heart, and kidney. Two novel imaging agents for Positron emission tomography PET have pre-clinically demonstrated promising target binding and organ distribution characteristics . However, the relevant disease monitoring in clinical setup would require multiple repetitive examinations per year.
Thus it is of paramount importance to investigate the absorbed doses and total effective doses, and thus potential maximum number of examinations per year.
Methods Two cyclic peptide analogues coupled via ethylene glycol linker to either 2- 4,7-bis 2- tert-butoxy -2oxoethyl -1,4,7-triazonanyl acetic acid NO2A-Col or 4- 4,7-bis 2- tert-butoxy oxoethyl -1,4,7triazacyclononanyl tert-butoxy oxopentanoic acid NODAGA-Col were labelled with 68Ga. Ex vivo organ distribution study was performed at 5, 10, 20, 40, 60, and min time points.
Results Ex vivo organ distribution revealed fast blood clearance half-life of 1. Organ absorbed dose for kidneys was 0. The total effective dose was 0. The statistically significant difference in the uptake of the agents in liver and spleen has been demonstrated previously  and was also reflected in this study with higher residence time and organ absorbed doses for [68Ga]Ga-NODAGA-Col.
Conclusions The total effective dose would allow for at least six examinations per year that might be sufficient for the adequate disease monitoring in longitudinal studies and routine clinical setup. Acknowledgements References  Velikyan et al. Labelling peptides with the Modular-Lab EAZY module is performed using an acetone-free method2 without purification of the product. From a radiopharmaceutical point of view the reagents and the cassettes need to be produced according to guideline ICH Q7A — GMP for APIs, since they contribute to the composition of the final product.
All reagents were supplied in GMP quality. Acknowledgements References 1Oxboel, J. The biological properties of the complexes were investigated by in vitro binding assays and uptake studies in DU and MCF-7 human prostate cancer and breast adenocarcinoma, respectively cell lines. The in vivo kinetics of the 99mTc-labeled radiotracer was investigated in ICR mice by biodistribution studies.
The log D value at pH 7. The uptake of [99mTc]6 in DU and MCF-7 cells was time-dependent and could be blocked by haloperidol dosedependently. Biodistribution studies in mice showed low uptake of [99mTc]6 in muscle at 4 h postinjection. Administration of haloperidol 5 min prior to injection of [99mTc]6 significantly reduced the radiotracer accumulation at 1 h and 2 h postinjection in the brain, heart, lungs, spleen, and kidney.
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Product may includes warranty, and accessories found with the original product.Page 5 Battery may toshiba satellite as charge while computer is consuming full power. Satellite AS Notebook psa70uowl00g. Francesconi2, Jason S. As potential sites for radiolabelling the aprotinin sequence contains four lysine residues, which do not contribute to the inhibitory properties .
Prepared characteristic representatives of all three groups are shown in Figure 1. Toshiba Satellite AS A70 Series drivers are tiny programs that enable your Laptop hardware to communicate with your operating system software. Therefore, we are investigating 89Zr-siderophores as a promising class of imaging agents for assessment of bacterial infection.
Louis, Missouri, United States Objectives Radiopharmaceuticals capable of imaging the functional transport activity of breast cancer resistance protein BCRP, ABCG2 are desired to assist in the stratification of chemotherapeutic choices.
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