A new polysaccharide macromolecular contrast agent for MR imaging: biodistribution and imaging characteristics

The aims of this study were to characterize certain physicochemical, pharmacokinetic, and enhancement properties of a new macromolecular contrast agent, carboxymethyl hydroxyethyl starch-(Gd-DO3A)(35) [CMHES-(Gd-DO3A)(35)], consisting of a polysaccharide backbone covalently derivatized with multiple macrocyclic chelating groups for gadolinium. CMHES-(Gd-DO3A)(35) has an average molecular weight of 72 kD and a plasma half-time of 8.4 hours. T1 and T2 relaxivities are 14.1 +/- 0.1 L mmol(-1) * sec(-1) and 17.8 +/- 0.9 L mmol(-1) * sec(-1), respectively, for each gadolinium ion measured at 39 degrees C and 20 Mhz; this T1 relaxivity is more than 4 times that of gadopentetate. Seven days after intravenous administration only relatively small amounts of gadolinium could be detected in blood or other tissues of rats. The compound was well tolerated in diagnostic dosages by all experimental animals. Magnetic resonance angiography performed within 1 hour of CMHES-(Gd-DO3A)(35) administration showed a near-constant and strong enhancement of blood in arteries and veins. Analysis of dynamic enhancement patterns of experimental tumors (MAT-LyLu prostate cancer implanted in rats) following intravenous CMHES-(Gd-DO3A)(35) administration yielded quantitative estimates of tumor plasma volume and microvessel permeability; the demonstrated hyperpermeability of tumor microvessels was easily distinguished from the absence of measurable microvascular permeability in non-neoplastic soft tissues.     

Improved imaging of infections by avidin-induced clearance of 99mTc-biotin-PEG liposomes.

This article describes the preparation and optimization of biotin-polyethyleneglycol (PEG) liposomes and their application in experimental infection models to improve the scintigraphic imaging of infection and inflammation. METHODS: Biotin was coupled to PEG-distearoylphosphatidylethanolamine (DSPE) and subsequently incorporated in the PEG liposomes. Biotinylated liposomes were radiolabeled with 99mTc-hydrazinonicotinamide. In vitro binding studies were performed to find the optimal biotin concentration in the liposomes. In rats the biodistribution of the 99mTc-biotin-PEG liposomes was compared with the biodistribution of normal (nonbiotinylated) 99mTc-PEG liposomes. Furthermore, in vivo studies in rats were performed to study both the effect of the biotin content and the optimal avidin dose for efficient clearance of the liposomes. Liposomes containing 0.5 or 1.0 mol% biotin-PEG-DSPE were compared in rats with a Staphylococcus aureus infection in the left calf muscle. Avidin was injected 4 h after injection of the liposomes. RESULTS: Biotinylation of the liposomes did not affect their in vivo behavior. All biotin-PEG liposome formulations tested showed good in vitro avidin binding with 50% inhibitory concentrations ranging from 36 to 8 micromol/L. With avidin doses higher than 100 microg, both preparations rapidly cleared from the circulation. As a result, abscess-to-blood ratios increased 5-fold. To illustrate the potential of the avidin-induced clearance of radiolabeled PEG liposomes, we also studied the 99mTc-biotin-PEG liposomes in rabbits with a subcutaneous S. aureus abscess. The infection was visualized only after injection of 100 microg avidin. CONCLUSION: This study shows that biotin-coated 99mTc-PEG liposomes in combination with the injection of avidin can lead to improved imaging of infection or inflammation localized especially in regions with high blood-pool activity.     

Biodistribution and clearance of contrast-carrying MREV liposomes

Contrast-carrying liposomes (CCLs) have been shown to increase the attenuation coefficient of the liver and spleen during computed tomographic (CT) imaging. A modification of the reverse-phase evaporation preparative technique ("MREV") led to production of CCLs that entrap contrast media efficiently. After intravenous injection, MREVs are phagocytized by the Kupffer cells of the liver and the macrophages of the spleen. The biodistribution and clearance of MREVs were studied to evaluate their potential for clinical use, MREVs carrying iodine-125-iotrolan were administered intravenously to 12 rats at doses of 400 mg lipid (containing 400 mg iodine) per kilogram body weight. Pairs of rats were anesthetized and scanned at 3 hours, and 1, 7, 14, 27 and 48 days; CT attenuation values of liver, spleen, blood, kidneys and bladder were measured. Immediately following CT, the rats were killed and tissue specimens were radioassayed. Maximum iodine content in liver and spleen was reached at one day and sustained at high levels for seven days. Biologic removal half-time of the agent in both the liver and spleen was six days. Maximum CT enhancements over baseline were observed at 24 hours and reached 210 delta HU in the liver and 880 delta HU in the spleen per gram iodine injected per kilogram body weight. It is concluded that MREVs have appropriate imaging characteristics, biodistribution, and clearance to be effective CT contrast agents.     

Synthesis, radiolabeling, biodistribution and fluorescent imaging of histidine-coupled hematoporphyrin

IntroductionHematoporphyrin (Hp) and hematoporphyrin derivatives (HpDs) have been widely used as photosensitizers in photodynamic therapy (PDT). Radiolabeling of HpDs is helpful for preclinical and clinical studies of PDT.MethodsThe histidine-coupled hematoporphyrin (His-Hp) was synthesized and radiolabeled with [^99mTc(CO)₃(H₂O)₃]⁺. Biodistribution of the radioligand and fluorescent imaging of His-Hp in mice bearing S180 tumor were investigated.Results[^99mTc(CO)₃]⁺-labeled His-Hp was electrically neutral, hydrophilic and stable. The biodistribution of the radioligand in S180 tumor-bearing mice was similar with that of nonlabeled HpD in the literature. The uptake of His-Hp in tumors and livers was confirmed by fluorescent imaging.ConclusionsThe complex [^99mTc(CO)₃]⁺–His-Hp might be suitable for in vivo dose evaluation of HpD in PDT.     

Hepatocyte-targeted nuclear imaging using 99mTc-galactosylated chitosan: conjugation, targeting, and biodistribution.

Galactosyl-methylated chitosan (GMC) is a galactosylated chitosan (GC) that is chemically modified to improve labeling efficiency with (99m)Tc compared with native GC. The aim of this study was to investigate the possibility of liver-targeted nuclear imaging with (99m)Tc-GMC bound to asialoglycoprotein receptors (ASGP-R). METHODS: GMC was obtained after the coupling of lactobionic acid, as the galactose moiety, and methyl iodide with chitosan. Using GMC-labeled fluorescein isothiocyanate (FITC-GMC), we examined whether GMC was localized in hepatocytes. After injection via the tail vein of mice with (99m)Tc-GMC and galactose-free (99m)Tc-methylated chitosan (MC), images were acquired with a gamma-camera equipped with a pinhole collimator. Biodistribution was obtained from 10, 60, and 120 min after injection. RESULTS: The composition of galactose groups in GC and tri-, di-, and monomethylated GC was confirmed by nuclear magnetic resonance spectroscopy. FITC-GMC was primarily positioned in hepatocytes, and not in Kupffer cells, of the mouse with a scattered pattern. The gamma-camera images showed rapid localization of (99m)Tc-GMC to liver. The percentage injected doses per gram (%ID/g) of liver were 11.155 +/- 2.332, 14.018 +/- 6.081, and 14.082 +/- 1.670 %ID/g (mean +/- SD) at 10, 60, and 120 min after injection, respectively. By contrast, galactose-free (99m)Tc-MC accumulated faintly in the liver. CONCLUSION: (99m)Tc-GMC specifically localized to the liver except for the kidneys in the mouse. GMC may be used to target the ASGP-R on the hepatocytes for nuclear imaging.     

Characterization of (111)In(3+) complexes of DTPA amide derivatives: biodistribution and clearance studied by gamma imaging

A large series of structurally related diethylenetriaminepentaacetic acid amide derivatives with different structures and lipophilic properties were synthesized and radiolabeled with (111)In(3+). Two of the more hydrophobic compounds studied ([(111)In]L(9) and [(111)In]L(10)) showed high affinity for human serum albumin (HSA). The biodistribution and clearance properties shown by all complexes upon injection in Wistar rats were followed by gamma imaging. The blood retention time of the chelates correlates better with their binding to HSA than with their hydrophilic/lipophilic ratio. Hydrophilic and negatively charged complexes undergo renal retention, while the majority of the lipophilic complexes are retained in the blood for a longer period of time and are cleared through the liver.     

99m Tc-glycopeptide: synthesis, biodistribution and imaging in breast tumor-bearing rodents.

This study was aimed to develop a glycopeptide (GP) to be used as a carrier for anti-cancer drug delivery. GP was synthesized by conjugating glutamate peptide and chitosan using carbodiimide as a coupling agent. Elemental analysis and capillary electrophoresis confirmed the purity was >95%. GP was labeled with sodium pertechnetate (Na99m TcO4) for in vitro and in vivo studies. Rhenium-GP was synthesized to support the binding site of 99m Tc at the glutamate positions 3-5. In vitro cellular uptake of 99m Tc-GP was performed in breast cancer cells. Cytosol had 60% whereas nucleus had 40% uptake of 99m Tc-GP. When cancer cells were incubated with glutamate or aspartate, followed by 99m Tc-GP, there was decreased uptake in cells treated with glutamate but not aspartate. The findings indicated that cellular uptake of 99m Tc-GP was via glutamate transporters. In addition, 99m Tc-GP was able to measure uptake differences after cells treated with paclitaxel. Biodistribution and planar imaging were conducted in breast tumor-bearing rats. Biodistribution of 99m Tc-GP showed increased tumor-to-tissue ratios as a function of time. Planar images confirmed that 99m Tc-GP could assess tumor uptake changes after paclitaxel treatment. In vitro and in vivo studies indicated that GP could target tumor cells, thus, GP may be a useful carrier for anti-cancer drug delivery.     

Contrast, resolution, and detectability in MR imaging

With the introduction of fast scan techniques and high field imagers, the ability to achieve very high resolution MR images in reasonable imaging times is now possible. Increased resolution allows for better detection of small, high contrast pathological features, but at some cost. Increasing resolution leads to a nonrecoverable decrease in signal-to-noise ratio per pixel and a loss of low contrast detectability for constant imaging time. This article examines the tradeoffs between image resolution, signal-to-noise ratio, and low contrast detectability in MR imaging. Contrast detail curves are presented for images collected in a constant imaging time, with constant field of view and bandwidth but at different resolutions, and these are compared with theoretical curves. The problem of measuring contrast levels in magnitude images, with different resolutions and receiver attenuation values, is discussed and a definition that accommodates these parameters developed. In addition, a clinical example is shown demonstrating a decrease in soft tissue differentiation with increasing resolution, again for fixed imaging time. The results indicate that moving to high resolution imaging matrices requires consideration be given to the sacrifice in low contrast detectability that occurs. Most importantly, it is shown that filtering a high resolution image to a lower resolution image, through nearest neighbor averaging, does not regain the detectability lost in initially collecting the high resolution image.     

A new myocardial imaging agent: synthesis, characterization, and biodistribution of gallium-68-BAT-TECH

In order to develop a new myocardial perfusion agent for positron emission tomography (PET), a new lipid-soluble gallium complex was evaluated. Synthesis, radiolabeling, characterization, and biodistribution of a unique gallium complex, [67Ga]BAT-TECH (bis-aminoethanethiol-tetraethyl-cyclohexyl), are described. The complex formation between Ga+3 and BAT-TECH ligand is simple, rapid, and of high yield (greater than or equal to 95%). This process is amenable to kit formulation. The complex has a net charge of +1 and a Ga/ligand ratio of 1:1. Biodistribution in rats shows high uptake in the heart as well as in the liver. When [68Ga] BAT-TECH was injected into a monkey, the heart and liver are clearly delineated by PET imaging, suggesting that this complex may be a possible tracer for myocardial perfusion imaging.     

Kinetics of avidin‐induced clearance of biotinylated bimodal liposomes for improved MR molecular imaging

Dual labeled liposomes, carrying both paramagnetic and fluorescent lipids, were recently proposed as potent contrast agents for MR molecular imaging. These nanoparticles are coated with poly(ethylene glycol) (PEG) to increase their blood circulation half‐life, which should allow extensive accumulation at the targeted site. To eliminate nonspecific blood pool signal from the MR images, the circulating liposomes should ideally be cleared from the circulation when sufficient target‐specific contrast enhancement is obtained. To that aim, we designed an avidin chase that allowed controlled and rapid clearance of paramagnetic biotinylated liposomes from the blood circulation in C57BL/6 mice. Avidin‐induced alterations in blood clearance kinetics and tissue distribution were studied quantitatively by determination of the Gd content in blood and tissue samples ex vivo. Intrinsic liposomal blood clearance showed bi‐exponential behavior with half‐lives t_1/2α = 2.1 ± 1.1 and t_1/2β = 15.1 ± 5.4 hours, respectively. In contrast, the contrast agent was cleared from the blood by the avidin infusion to <1% of the initial dose within 4 hours. Avidin‐induced liposomal blood clearance was also demonstrated in vivo by dynamic T₁‐weighted MRI. The ability to rapidly clear circulating contrast agents opens up exciting possibilities to study targeting kinetics, to increase the specificity of molecular MRI and to optimize nanoparticulate contrast agent formulations. Magn Reson Med 60:1444–1456, 2008. © 2008 Wiley‐Liss, Inc.     

99mTc demotate 1: biodistribution and elimination characteristics in rats

BACKGROUND AND METHODS: In this study, we investigated the biodistribution and elimination characteristics of a new radiolabelled somatostatin analogue, 99mTc demotate 1, in rats by in-vivo biodistribution and elimination experiments, perfused rat liver and kidney experiments and micro-autoradiography of renal tissue. RESULTS: Rapid clearance from blood and most organs was found. High and long-term uptake in organs with high density of somatostatin receptors (the adrenals and pancreas) and in stomach and intestine was reduced in non-radiolabelled octreotide pretreated animals. The predominant urine excretion was associated with an accumulation of 99mTc demotate 1 in the kidney, mainly in the renal cortex. This uptake was not affected by non-radiolabelled octreotide pretreatment. CONCLUSION: 99mTc demotate 1 is a prospective radiopharmaceutical for use in human medicine in somatostatin receptor-positive tumour imaging and its potential should be confirmed in further experiments and clinical trials.     

Synthesis, characterization, and biodistribution of [113mIn]TE-BAT: a new myocardial imaging agent

In order to develop a new myocardial perfusion agent, new lipid-soluble complexes containing a net charge of +1 were evaluated. Synthesis, radiolabeling, characterization, and biodistribution of a unique indium complex, [113mIn]TE-BAT (tetraethyl-bis-aminoethanethiol), are described. The complex formation between In +3 and TE-BAT ligand is rapid, simple, and of high yield (greater than or equal to 95%). This process is amenable to kit formulation. The complex has a net charge of +1 and an In/ligand ratio of 1:1. Biodistribution in mice shows higher heart uptake and longer retention as compared to 201TI. This complex, when labeled with 111In, shows promise as a possible tracer for myocardial perfusion imaging.     

Characterisation of 67Ga3+ complexes of triaza macrocyclic ligands: biodistribution and clearance studies

The 67Ga3+ complexes of three triazamacrocycles, 1,4,7-triazacyclononane-N,N',N'-triacetic acid (NOTA), its phosphonate analog 1,4,7-triazacyclononane-N,N',N'-tris(methylenephosphonic) acid (NOTP), and the monoethyl ester of NOTP, 1,4,7-triazacyclononane-N,N',N'-tris (methylenephosphonate-monoethylester) (NOTPME) were studied for possible use as radiopharmaceuticals. Biodistribution studies and gamma imaging were performed in Wistar rats. The present results demonstrated that all the macrocyclic complexes studied display renal clearance and are almost completely eliminated within 24 h. The [67Ga](NOTP)3- chelate, with a large negative charge, has a considerably slower uptake and elimination by the kidneys than the neutral [67Ga](NOTA) and [67Ga](NOTPME) chelates. We have thus demonstrated a charge-clearance relationship for a series of stable and well characterized complexes. The high stability and rapid renal excretion properties displayed by the NOTA and NOTPME chelates support their possible application as imaging agents for kidney structural and functional studies.     

99mTc-Daunorubicin a potential brain imaging and theranostic agent: synthesis,quality control,characterization, biodistribution and scintigraphy

Daunorubicin is a chemotherapeutic antibiotic of the anthracycline family used for the treatment of many type of cancers when doxorubicin or other less effective drugs cannot be used. The aim of the present study was labeling of Daunorubicin with ^99mTc, quality control, characterization, and biodistribution of radiolabeled Daunorubicin. Labeling efficiency was determined by ascending paper chromatography. All the experiments were performed at room temperature (25 °C ± 2 °C). More than 96% labeling efficiency with ^99mTc was achieved at pH 5–6, 2–4 μg stannous chloride and 300 μg of ligand in few minutes. The characterization of the compound was performed by using HPLC, electrophoresis and shake flask assay. Electrophoresis indicates that Tc-99 m-Daunorubicin is neutral, HPLC confirms the single specie of the labeled compound, while shake flask assay confirms high lipophilicity. The biodistribution studies of ^99mTc-Daunorubicin were performed in rats. Significantly higher accumulation of ^99mTc-Daunorubicin was seen in brain of normal rats. Scintigraphy was also indicating higher accumulation of ^99mTc-Daunorubicin in brain of normal rabbits.     

Radioiodination of glycoprotein-conjugated liposomes by using the Bolton-Hunter reagent and biodistribution in tumor-bearing mice

We have developed a suitable radiolabeling method for our new type of glycoprotein-liposome conjugate (GCL), in order to investigate its potential utility as a drug carrier that can target the cellular functions of carbohydrate-binding proteins. In order to obtain radiolabeled GCL with high labeling efficiency, we introduced p-hydroxyphenylpropyl groups into the liposome membrane through the amine moiety of a constitutive phospholipid, dipalmitoylphosphatidylethanolamine (DPPE) by using Bolton-Hunter reagent (BHR). Radioiodination of the introduced tyrosyl groups was performed by the Chloramine-T method. The labeling efficiency of the BHR-treated liposome conjugate was high in comparison with that of the BHR-untreated liposome conjugate. An in vitro inhibition study showed that the binding affinity of 125I-labeled BHR-treated GCL (125I-F3S-BH) with lectin was twice as high as that of untreated conjugate (125I-F3S). The biodistribution of 125I-F3S-BH in mice was considerably different from that of 125I-F3S. 125I-F3S-BH was more rapidly taken up by the liver and was more rapidly excreted from the liver than 125I-F3S. Moreover, 125I-F3S-BH accumulated more rapidly into the kidneys, which resulted a lower radioactivity in the blood circulation at an earlier time point than in the case of 125I-F3S. The characteristics of tumor accumulation of 125I-F3S-BH and 125I-F3S were similar to those in blood. If F3S is to be employed as an in vivo targeting ligand in biodistribution studies, BHR would be a suitable tool for radiolabeling because it allows GCL to retain the biological activity and characteristics of the unmodified conjugate.     

Diverse characteristics of 111In labelled anti-CEA monoclonal antibodies for tumour immunoscintigraphy: radiolabelling, biodistribution and imaging studies in mice with human tumour xenografts

Three monoclonal anti-CEA antibodies, designated 161, 198 (both IgG1) and 228 (IgG2a) have been labelled with 111In via DTPA chelation and assessed for localization in human gastro-intestinal carcinomas as xenografts in athymic nude mice. Following reaction of the antibodies with DTPA anhydride, efficiency of chelation of 111In varied between the antibodies with mean values of 30%, 52% and 62% with 161, 198 and 228 respectively. Gel filtration chromatography with all three labelled antibodies showed radiolabel predominantly coincident with IgG with little radioactivity in either high molecular weight form or as free 111In. However, the efficiency of binding of radiolabelled antibodies to CEA producing tumour cells varied, with maxima of 42%, 65% and 20% for 161, 198 and 228. In vivo, in mice, 111In was excreted at virtually identical rates (half times approx. 12 days) with all three preparations and this was similar to the clearance of indium injected as 111In-indium chloride, but 111In-DTPA was rapidly eliminated (half time approximately 5 h). After injection into mice with CEA producing xenografts of colon carcinoma HT29 and LS174T and gastric carcinoma MKN 45, circulating radiolabel was still predominantly in the form of labelled antibody with little or no detectable immune complexes or 111In labelled transferrin. Tumour localization of all three antibodies was visualized by gamma camera imaging with target: non target ratios of up to 5:1. Dissection of mice with MKN45 gastric carcinoma xenograft showed 16%, 19.5% and 13% of the injected dose of 111In from 161, 198 and 228 antibodies in each g of tumour tissue.(ABSTRACT TRUNCATED AT 250 WORDS)