Comparison of five bifunctional chelate techniques for 90Y-labeled monoclonal antibody CO17-1A.

Monoclonal antibody CO17-1A was radiolabeled with 90Y by five bifunctional chelate techniques. Radiation absorbed dose estimates for normal organs and tumor were calculated for each preparation based on timed tissue distribution studies in nude mice bearing SW 948 human colorectal carcinoma xenografts. The cyclic DTPA anhydride technique was inferior to the four other techniques studied. Data for SCN-Bz-DTPA and SCN-Bz-Mx-DTPA, which were conjugated to epsilon-lysyl amino groups, were similar to those for NH2-Bz-DTPA and NH2-Bz-Mx-DTPA, which were conjugated site specifically to oligosaccharides.

Radioimmunotherapy of human colorectal carcinoma xenografts using 90Y-labeled monoclonal antibody CO17-1A prepared by two bifunctional chelate techniques.

Monoclonal antibody CO17-1A, which has specificity for colorectal and pancreatic carcinomas, was radiolabeled with the pure beta emitter, 90Y, by either the cyclic diethylenetriaminepentaacetic acid (DTPA) anhydride technique or by a site-specific bifunctional chelate technique using 1-(p-aminobenzyl)DTPA (p-NH2-Bz-DTPA). Female nude mice bearing SW 948 human colorectal carcinoma xenografts were given injections i.v. of 90Y-labeled monoclonal antibody CO17-1A at dosages of 100, 150, and 200 muCi/25 g body weight. Unlabeled CO17-1A (100 micrograms/25 g body weight) was coadministered. In animals receiving 90Y-CO17-1A prepared by the cyclic DTPA anhydride technique, tumor volume was unchanged from base line at a dose of 200 microCi/25 g. As the dosage of 90Y-CO17-1A increased, the rate of tumor growth decreased, but all experimental animals in this group died between 14 and 21 days. In contrast, CO17-1A radiolabeled with 90Y by the site-specific p-NH2-Bz-DTPA bifunctional chelate technique produced a maximum tumor volume reduction of 87% in the 200 microCi/25 g group by day 15, and no deaths were noted in any of the 90Y-CO17-1A-treated groups for 71 days. Dose-response curves again showed increased tumoricidal effects with increased dosages of 90Y-CO17-1A. S-2-(3-Aminopropylamino)ethylphosphorothioic acid, commonly known as WR-2721, is a radioprotective drug which has been shown to protect against bone marrow depression in irradiated humans. No protection was observed when WR-2721 was used as an adjunct to treatment with 90Y-CO17-1A prepared by either the cyclic DTPA anhydride technique or the site-specific p-NH2-Bz-DTPA technique. When the site-specific p-NH2-Bz-DTPA technique was used, the reduction in WBC and hemoglobin levels correlated with increasing bone marrow toxicity at higher doses. We conclude that CO17-1A labeled with 90Y via the site-specific p-NH2-Bz-DTPA technique has potential for radioimmunotherapy of human colorectal carcinoma.

90Y-labeled monoclonal antibodies for cancer therapy.

Monoclonal antibody 17-1A, which has specificity for colorectal carcinoma, was labeled with 90Y (10-20% radiolabeling yield). Tissue distribution studies in tumor-bearing nude mice were carried out. 90Y-labeled 17-1A showed good uptake in the SW 948 colon carcinoma cell line. However, 90Y-labeled A5C3, a monoclonal antihepatitis virus antibody studied as a control, showed similar uptake in this tumor. Neither antibody was taken up well by a WM-9 melanoma. It is believed that the loss of specificity observed is due to the low specific activity of the 90Y-labeled monoclonal antibody preparations used. This hypothesis is supported by radioimmunoassay data.

Comparison of 68Ga-EDTA, [1-11 C]alpha-aminoisobutyric acid, and [99mTc]sodium pertechnetate in an experimental blood-brain barrier lesion.

Unilateral opening of the blood-brain barrier (BBB) in rats, induced by infusion of a hyperosmotic solution of mannitol into the left internal carotid artery, was applied to compare 68Ga-EDTA, [1-11C]alpha-aminoisobutyric acid (using the long-lived 14C-labeled analog), and [9mTc]sodium pertechnetate as agents for diagnosis of BBB disruption. Of the three agents and two time intervals studied, 68Ga-EDTA at 30 min postinjection gave the highest target-to-nontarget ratio. In addition, 68Ga-EDTA, unlike commonly used [99mTc]sodium pertechnetate, can be used in conjunction with positron emission tomography, which could make possible earlier and better assessment of BBB defects using 68Ga-EDTA.

Liposomal preparations of calcium- or zinc-DTPA have a high efficacy for removing colloidal Ytterbium-169 from rat tissues.

Results from this study demonstrate that liposomes increase the effectiveness of chelating agents in removing heavy metals from contaminated tissues in vivo. We compared the ability of free and liposomal preparations of Ca- and Zn-diethylenetriaminepentaacetic acid (DTPA) to remove 169Yb from tissues of rats previously injected intravenously with either soluble or colloidal forms of 169Yb . Although single injections of liposomal Zn-DTPA were better than the free chelator in reducing the body burden of 169Yb administered in a soluble form (citrate), the liposomal preparations of Zn-DTPA or Ca-DTPA were even more effective in removing the 169Yb that had been injected as the colloidal form. However, a second injection of liposomal Zn-DTPA given 8 days after the initial treatment was not as effective in removing 169Yb as a second injection of the free chelator. Whether injected in the free or liposomal forms, Ca-DTPA was more effective than Zn-DTPA in removing the colloidal 169Yb . Significantly lower amounts of colloidal 169Yb remained in the liver, kidney, muscle, bone, and blood of rats after injection of the liposomal preparations of Ca- or Zn-DTPA than in the corresponding organs of the controls (P less than 0.01). The liposomal preparations were also more efficient in reducing the retention of colloidal 169Yb in bone and blood than the free chelators (P less than 0.01).

Production of L-[1-11C]valine by HPLC resolution.

Based on a recently developed analytical technique, preparative high-performance liquid chromatographic (HPLC) resolution of DL-[1-11C]valine has been achieved. A conventional reverse-phase HPLC column and a chiral mobile phase (aqueous solution of L-proline, cupric acetate, and sodium acetate) were used. The copper can be removed from the L-valine fraction by precipitation as the sulfide, and final purification by cation-exchange chromatography yields L-[1-11C]valine in a form that is acceptable for clinical positron tomographic studies. This purification method does not remove the L-proline introduced in the resolution process, but added L-proline did not affect the tissue distribution of L-[1-14C]valine in rats. We have produced up to 60 mCi of L-[1-11C]valine in an overall synthesis and resolution time of 50 min. This procedure should be adaptable to the rapid resolution of other C-11-labeled amino acid racemates.

Studies of the vivo uptake of Ga-67 by an experimental abscess: concise communication.

The blocking of Ga -67 plasma protein-binding sites-by administration of scandium citrate, ferric citrate, and a colloidal hydrous ferric oxide preparation-reduced the uptake of Ga-67 in normal soft tissues and also that in the viable portion of an experimental abscess. On the other hand, enhancement of Ga-67 plasma protein binding by administration of rabbit apotransferrin increased Ga-67 uptake in both abscess and normal soft tissues. These results indicate that the pathways of Ga-67 from blood into inflammatory processes and normal soft tissues may be similar. However, when Ga-67 plasma protein binding was increased by inducing anemia, a markedly decreased Ga-67 uptake in the abscess resulted, whereas uptake in normal soft tissue was still elevated. It is possible that the discrepancy between the effects of apotransferrin and anemia on abscess-tissue uptake of Ga-67 resulted from a secondary effect produced by anemia, i.e., a decrease in the macrophage population in the abscess. Taken as a whole, the results obtained suggest that Ga-67 leaves the blood and enters inflammatory lesions by pathways that are probably quite different from those in a soft-tissue tumor, and that the routes for abscesses may be similar to those occurring in normal soft tissues.

Studies of the in vivo entry of Ga-67 into normal and malignant tissue.

Previous studies of the effect of scandium on the tissue distribution of Ga-67 suggest that Ga-67 makes its initial in vivo entry into normal and malignant tissues by different routes. (Scandium blocking of plasma protein Ga-67 binding increased Ga-67 excretion, decreased its uptake in normal tissues, but had little effect on rodent tumors.) In further studies we have used other methods to alter the plasma binding of Ga-67. Iron saturation of plasma produced effects on Ga-67 tissue distribution similar to those observed with scandium. On the other hand, increasing Ga-67 plasma binding through induction of anemia and administration of apotransferrin produced the reverse of the effects observed with scandium and iron. We conclude that the initial in vivo entry of Ga-67 into tumor tissue involves mainly an unbound or loosely bound form of Ga-67, whereas its uptake by normal soft tissues is strongly promoted by its binding to transferrin.

The effect of scandium on the tissue distribution of Ga-67 in normal and tumor-bearing rodents.

In rats and mice the intravenous administration of scandium before or with Ga-67 produces an increase in Ga-67 excretion and bone deposition, coupled with pronounced decreases in the uptake of Ga-67 in soft tissues. These effects result from the blocking by scandium of Ga-67 plasma-protein binding sites, which forces Ga-67 into an unbound or loosely bound state. This increases Ga-67 excretion and bone deposition, which in turn acts to produce greatly reduced Ga-67 uptake in soft tissues. When tumor-bearing rats and mice are administered scandium, similar effects occur, but the uptake of Ga-67 by tumor tissue remains unchanged. This suggests that Ga-67 enters tumor and normal soft tissues by different routes. With tumor, an unbound or loosely bound form of gallium is primarily involved, whereas with normal soft tissues this route is apparently of minor importance.

Effect of food intake on the tissue distribution of gallium-67: concise communication.

Fasting affects the body retention and tissue distribution of Ga-67 in experimental animals. In Ga-67 experiments, therefore, a difference in food intake between treated and control animals might result in confusing side effects. We have observed this in irradiation studies. It is suggested that a fasting regimen should be imposed in any Ga-67 animal study where an alteration in food intake might be experienced in the treated group.

DL-[Carboxyl-11C]tryptophan, a potential agent for pancreatic imaging; production and preclinical investigations.

In animal studies, DL-[carboxyl-14C]tryptophan [DL-Try(C-14)] showed a high specificity for the pancreas, which suggested the potential of DL-[carboxyl-11C]tryptophan [DL-Try(C-11)] for clinical pancreatic inaging. The blood clearance and tissue uptake of the amino acid were very rapid, and no carrier effect was observed through a dose of 5 mg/kg. None of three transplanted hamster pancreatic adenocarcinomas that we studied showed a selective uptake of DL-Try(C-14) by the tumor, and none of the three enzymatic regimens investigated gave significant enhancement of the pancreatic specificity. Commercial L-Try(C-14) gave slightly better pancreatic specificity than the analogous racemic compound but without enough improvement to warrant attempts at optical resolution. DL-Try(C-11) was synthesized in amounts up to 325 mCi using a rapid, high-temperature, high-pressure modification of the Bücherer-Strecker amino acid synthesis. Yields ranged from 30--60%, and a total of 40 min was required for synthesis and chromatographic purification. DL-Try(C-11) thus appears to have significant potential as a clinical pancreas-imaging agent, particularly when used in conjunction with positron computerized transaxial tomography.

A quantitative study of the subcellular localization of 67Ga.

This paper describes a quantitative procedure for the isolation of 67 Ga-binding granules (GBG) from normal rat liver and Morris 5123C hepatoma homogenates by a combination of rate and isopyknic density gradient zonal centrifugation. Another class of GBG has been found that is much smaller than the lysosomal GBG we have previously described. These smaller particles, or microvesicles, bind the largest portion of the 67Ga found in the hepatoma whereas, in the liver, the GBG lysosomes are the major binding component. Previously, we had shown that considerably more 67Ga is taken up in hepatoma than in liver (as percentage of administered dose per g of tissue). The preferential association of 67Ga with these microvesicles in the 5123C hepatoma may be indicative of a basic difference between normal and malignant tissue.

Gallium-67 localization in rat and mouse tumors.

Various nonosseous tumors in mice and rats concentrate gallium-67-ESR-586 preputial gland carcinoma, P-1798 lymphosarcoma, C3H/ HeJ mammary tumor, AKR/J thymic lymphoma, R-3259 giant cell sarcoma, Walker-256 carcinosarcoma, and a new transplantable neoplasm of rats. This last tumor showed the greatest affinity for galliun. Viable tumor cells are mainly responsible for uptake of gallium-67; autoradiographic studies indicate it is chiefly located in tumor cytoplasm.