Izumi Ogihara-Umeda

Active removal of radioactivity in the blood circulation using biotin-bearing liposomes and avidin for rapid tumour imaging

In order to shorten the delay between the administration of tumour-imaging agents and obtainment of scintigrams, rapid delivery of radionuclide to tumours followed by rapid clearance from the blood is required. We used liposomes with biotin bound on their surface (B-liposomes) as carriers for rapid delivery. For rapid blood clearance, we employed avidin in the expectation that the strong affinity between biotin and avidin would result in the aggregation of B-liposomes in the blood circulation, and that these aggregates would be taken up rapidly by the reticuloendothelial system, resulting in the rapid elimination of liposomes and the radionuclide encapsulated in them. When B-liposomes encapsulating gallium-67 deferoxamine were intravenously administered to mice bearing sarcoma 180, large amounts of 67Ga were delivered to tumours by 4 h after injection, though the 67Ga blood level remained high. On the other hand, administration of avidin 4 h after administration of the B-liposomes dramatically reduced the blood level so that it was only 5% of that in the non-treated group 1 h later. As a result, the tumour-to-blood ratio reached nearly 14 at 5 h after radionuclide administration, suggesting that rapid tumour-imaging will be feasible by means of this method.

Cholesterol enhances the delivery of liposome-encapsulated gallium-67 to tumors

The effect of incorporation of cholesterol (CH) into liposome membranes on the delivery of 67Ga encapsulated in liposomes to tumors was studied. The changes of the blood clearance of liposomes, liposome, stability in serum and liposome uptake by the liver and spleen were also examined. Liposomes were prepared from distearoylphosphatidylcholine with various amounts of CH. It became clear that large amounts of CH (above 33 mol%) dramatically enhanced liposomal delivery of 67Ga to sarcoma 180 solid tumor in mice. Large amounts of CH increased the liposome stability in serum and decreased liposome uptake by the liver and spleen after intravenous injection, thus prolonging the blood clearance of liposomes. These observations suggested that the large delivery of 67Ga by CH-rich liposomes resulted from the extended retention and increased amount of liposomes in the circulation, caused by the incorporation of large amounts of CH. Small amounts of CH decreased liposome stability and hastened blood clearance, but had little effect on 67Ga delivery to the tumor. CH-rich liposomes showed high tumor uptake, with high tumor to blood and tumor to tissue ratios, of 67Ga. It is anticipated that 67Ga-carrying liposomes will be an excellent tumor imaging agent for clinical use, provided that a correct choice of CH content is made.

Development of a liposome-encapsulated radionuclide with preferential tumor accumulation—the choice of radionuclide and chelating ligand

Various radionuclide-ligand complexes were encapsulated in liposomes and their accumulations in tumors were studied. Increased tumor accumulation was observed with every complex in the liposome-encapsulated form. However, different accumulation levels were registered for the various radionuclides even though they were all delivered using a similar liposome formulation. Though the liposomes remained intact in the circulation, they were degraded in the tumor, liver and spleen eventually. Thus, this suggests that tumor accumulation of liposome-encapsulated radionuclides is dependent on not only the in vivo behavior of the liposomes themselves, but also the characteristics of nuclide-ligand complexes after their release from liposomes. A correct choice of radionuclides and ligands for encapsulation in liposomes would enable excellent tumor-imaging agents to be achieved.

Preventive effect of pyrroloquinoline quinone (PQQ) on bilivedrin accumulation of the liver of chick embryo after glucocorticoid administration

Administration of a high dose of hydrocortisone hemisuccinate sodium (0.25 mumol/egg) to 15-day-old fertilized hens eggs caused accumulation of biliverdin in the embryonic liver. The responses were 10 to 20-fold higher than controls within 48 hr as a result of stimulation of biliverdin synthesis in the liver and the decrease of biliverdin excretion from the liver. This accumulation was effectively prevented by PQQ, possibly through the enhancement of biliverdin excretion from the liver to the gallbladder. This PQQ action was due to its preventive effect against the decrease of glutathione in the liver caused by glucocorticoid since glutathione is suggested to play a role in elimination of bile components from the liver.

Involvement of hepatic glucocorticoid receptor-mediated functions in steroid-induced cataract formation

Determination of whether the steroid-induced cataract formation is caused through glucocorticoid (GC) receptor-mediated process was conducted by using GC antagonist (RU486) and anti-GC receptor antibody, and by sucrose density gradient ultracentrifugation analysis. (1) When 15 day-old chick embryos were treated with dexamethasone (DEX, 0.025 micromol per egg), their lenses started to form an opaque ring around the peri-nuclear region (stage II-III) after 12 hr and developed into nuclear-like cataract (stage IV-V) after 44 hr. The cataract formation examined at the 44 hr could be effectively prevented by administration of RU486 (0.2 micromol per egg) ranging from 2 hr before to 12 hr after the DEX administration. (2) GC receptor was present in liver, but could not be determined in lens by western blot analysis using monoclonal anti-GC receptor antibody. (3) Sucrose gradient ultracentrifugation analysis indicated that the receptor (9S) in the liver could be transformed to the 4S form after 0.4M NaCl treatment. Combined with our previous data, this suggests that changes in hepatic functions mediated by the GC receptor after the GC administration may be involved in the process of the cataract formation.

Sequential enzymatic synthesis and biodistribution of radiolabelled inositol and inositol analogs

The desired radiolabelled inositol (Ins) and inositol-1-phosphate (IP) were obtained from radiolabelled glucose by sequential enzyme reactions in a short time. Rapid separation and fractionation of enzymes and labelled products from the reaction mixture was achieved by HPLC with a gel-permeation chromatography column. Examination of the biodistribution of radiolabelled Ins, IP and their acetylated analogs suggested that intact Ins labelled with 11C would be more effective than 11C-labelled acetylated Ins as a brain diagnostic agent for positron emission tomographic studies of the metabolism of phosphatidylinositol and its role as a second messenger in the brain.