Distribution of [14C]suramin in tissues of male rats following a single intravenous dose.

PURPOSE: Suramin has been shown to have efficacy in treatment of prostate cancer. In the present study we evaluated distribution of [14C]suramin in tissues over time following a single intravenous dose. METHODS: Male rats were given a single IV dose of 300 mg/kg [14C]suramin and sacrificed at 1 or 6 hours, or at 1, 7, 14, 28, 56, or 84 days postdose. Radioactivity remaining in tissues was measured by quantitative whole body autoradiography. RESULTS: At one hour highest tissue activity was found in blood vessel walls and caecum, followed by lung, blood, skin, preputial, thyroid, brown fat, heart, kidney, lymph nodes, liver, salivary, adrenal, Harder's and lacrimal glands, prostate, and spleen. Considerable activity was present in membranes surrounding muscle groups, bone and other organs. Relatively low activity was found in brain tissue although persistent concentration was evident in choroid plexus. High levels were present in bladder and caecum contents. Activity declined in blood but continued to increase in many tissues at later time points. Kidney reached maximum levels at 7 days postdose and retained concentration considerably higher than other tissues over the course of the study. Concentrations in tissues were persistent and considerable activity remained at 84 days postdose. Terminal elimination half life in tissues was prolonged, approximately 39 days in blood and 91 and 102 days in kidney and spleen, respectively. Uptake in prostate was highest in membranous structures separating secretory lobules. CONCLUSION: Suramin is widely distributed to tissues and appears to have particular affinity for boundary membranes surrounding organs and other structural tissue elements, possibly due to uptake by glycosaminoglycans. Antitumor activity may be related to inhibition of growth factors associated with these elements.

Distribution of tacrine and metabolites in rat brain and plasma after single- and multiple-dose regimens. Evidence for accumulation of tacrine in brain tissue.

Tacrine [1,2,3,4-tetrahydro-9-acridinamine monohydrochloride monohydrate (THA), Cognex] is a potent acetylcholinesterase inhibitor recently approved for treatment of mild-to-moderate Alzheimer's disease. The potential for THA and/or a metabolite of THA to accumulate in brain tissue was investigated by autoradiographic and metabolic profiling techniques in rats given single and multiple doses of [14C]THA. In addition, the brain-to-plasma distribution time course of orally administered 1-hydroxy-THA (1-OH-THA, 24 mg/kg), a primary rat metabolite with anticholinesterase activity, was also examined. Results from a 16 mg/kg single-dose study showed THA to cross the blood-brain barrier readily and concentrate in brain tissue, approximately 5-fold compared with plasma. The metabolite 1-OH-THA was found in much lower amounts relative to THA and when given separately at a similar dose the levels in brain tissue were comparable with plasma concentrations. After multiple-dose administration, THA concentrations in brain tissue were approximately 3-fold higher than those achieved after a single oral dose. However, concentration of 1-OH-THA metabolite increased only 50%. These data suggest a marked difference between the ability of THA and 1-OH-THA to accumulate in brain tissue and may reflect differences in lipophilicity as estimated by calculated log p values. The relevance of THA accumulation in brain tissue to delays observed in THA clinical management of Alzheimer's disease remains to be established.

Disposition of gabapentin (neurontin) in mice, rats, dogs, and monkeys.

Gabapentin, an analog of gamma-aminobutyric acid, exhibits anticonvulsant properties in both animal models and humans. Gabapentin pharmacokinetics was studied in laboratory animals using HPLC and radiometry. Oral bioavailability was 40% in monkeys administered 25 mg/kg, 79% in mice and rats receiving 50 mg/kg, and 80% in dogs administered 50 mg/kg. Binding to plasma proteins was < 3%. Maximum blood or plasma concentrations generally occurred within 2 hr of an oral dose. In rats and monkeys, increases in maximum plasma concentrations and/or areas under the curve were less than dose-proportional following oral administration, most likely because of saturable absorption. However, intravenous pharmacokinetics in rats were linear over the dosage range of 4-500 mg/kg. Mean intravenous elimination half-life was 1.7 hr in rats, 2.9 hr (14C only) in dogs, and 3.0 hr in monkeys. In rats and dogs, repeated administration did not alter gabapentin or 14C pharmacokinetics. Additionally, gabapentin did not induce hepatic cytochrome P450 monooxygenases in rats. There were no age- (rats only) or gender-associated changes in pharmacokinetic parameters. [14C]Gabapentin was extensively distributed to tissues. In the dog, gabapentin was metabolized to N-methylgabapentin (approximately 34% of dose); whereas metabolism in mouse, rat, and monkey was minimal (< 5%). The principal route of excretion was via urine. In summary, as an antiepileptic drug, gabapentin exhibited desirable pharmacokinetic properties, such as linear elimination kinetics, not highly bound to plasma proteins, not extensively metabolized, and not an inducer of hepatic cytochrome P450.

Quantitative whole-body autoradiographic analysis of the tissue distribution of orally-administered [14C]cholesterol in hypercholesterolemic rats.

The cholesterol-fed rat model has been used to examine the distribution of radiolabeled cholesterol by whole-body autoradiographic and quantitative videodensitometric methods. Animals were fed a hypercholesterolemic diet for 7 days, and were subsequently killed at 3, 6, 12, 24, or 72 h following a single oral dose of [14C]cholesterol. Maximum blood and tissue levels were observed at 12 h, while liver and adrenals were the most intensely labeled tissues. Liver maintained consistently high levels over the course of the study, while activity in other tissues declined moderately by 72 h, indicating the long half-life of cholesterol radioequivalents in tissue. The results of these experiments suggest that autoradiographic examination of cholesterol distribution in animals treated with pharmaceutical agents designed to modify cholesterol absorption or clearance will be useful in providing supplemental or confirmatory information on the drugs' mode of action.

Whole-body autoradiographic localization of [3H]phencyclidine and its metabolites in mice.

When evaluated by whole-body autoradiography (WBAR) and quantitative densitometry, [3H]phencyclidine (PCP) equivalents were found to be removed rapidly from blood, after a single iv dose in mice, and avidly taken up as early as 1 min after dosage by glandular tissues including thyroid, salivary glands, pancreas, pituitary and, most prominently, by stomach mucosa. Stomach:blood [3H]PCP concentration ratios showed that rapid secretion of [3H]PCP from mucosa to the stomach contents occurred within 2 min after dosing. During early intervals, chromatographic analysis of tissue sections demonstrated that PCP was present in brain, liver, and gut primarily in its unaltered chemical form. Mice killed at 60 and 120 min showed persistently high levels of [3H]PCP equivalents within the stomach and intestines, these levels being the highest of all other tissues densitometrically measured. The early time course and magnitude of [3H]PCP uptake by stomach glandular mucosa strongly suggests that cycling of PCP occurs principally through gastroenteric recirculation. Very striking was the high concentration of [3H]PCP radioactivity observed within the adrenal as early as 5 min. The concentration of [3H]PCP equivalents in pituitary, choroid plexus, cortex, hippocampus, and thalamus was highest at 1-20 min following injection. Application of high-resolution quantitative WBAR was found to be a useful tool in the study of the biodistribution of labeled PCP, especially during very early post-treatment time points where alternative tissue counting techniques would not be feasible.

Regional distribution of lead in the brains of lead-intoxicated adult mice.

The susceptibility of adult neural tissues to the detrimental effects of Pb poisoning has prompted the present distributional analysis of lead in the brains of chronically lead-exposed mice. A high-resolution microPIXE method was developed for measuring Pb in whole-brain cryosections derived from chronically lead-exposed mice. Spatial resolutions as small as 20 micron were obtained. Details of the methodology are presented together with procedures for Pb standard preparation and control measures employed to reduce potential errors of Pb assay associated with taking brain sections with steel alloy knives. The unique advantages of making microPIXE Pb determinations in nonpreselected brain anatomic regions using freeze-dried semithick cryosections are reviewed. The study revealed that, in lead-intoxicated mice, there existed wide regional variation in Pb concentration in the ppm range, in 50 micron sagittal or coronal sections. Higher Pb levels were found in discrete brain regions of lead-treated adult mice than in matched control brains. Suggestions for further studies of Pb kinetics using microPIXE methods in adults and immature animals, including components of neural barrier tissues, are reviewed.

Quantitative whole-body autoradiography of radiolabeled antibody distribution in a xenografted human cancer model.

Quantitative whole-body autoradiography (WBAR) was used to study the biodistribution of goat anti-carcinoembryonic antigen and normal goat IgG, each labeled with 125I, in hamsters bearing the carcinoembryonic antigen-producing GW-39 human colonic carcinoma xenograft. Comparisons between computer-assisted videodensitometric profiles of WBARs and tissue radioactivity counts were made at 1, 3, and 7 days following administration of the radiolabeled IgGs. The results indicated that maximal tumor accretion of the radiolabeled antibody and normal IgG occurred within 1-3 days, with a marked selective accretion of antibody in the tumor being evidenced at 3-7 days because of clearance of normal IgG. Radioactivity derived from antibody IgG showed 6.5 to 118.7 times that found in other tissues, as measured by videodensitometry, whereas organ radioactivity counting revealed ratios of only 6.7 to 29.6. Specificity of tumor-cell accretion of the radiolabeled antibody was confirmed by microscopic autoradiography, showing intense labeling of the proliferating perimeters of GW-39 tumors. WBAR was found to have a resolution of 0.10 to 0.25 mm in 100-g hamsters, which appears to be greater than the resolving power of external body imaging by gamma camera scintigraphy. These studies suggest the use of WBAR and microautoradiography to complement external imaging methods for the analysis of antibody distribution and localization in cancer radioimmunodetection models.

Quantitative autoradiography with radiopharmaceuticals, Part 2: Applications in radiopharmaceutical research: concise communication.

We describe the application of macroautoradiography, a relatively simple, quantifiable method for the evaluation of positron-emitting and gamma-emitting radiopharmaceuticals. We have investigated the response properties of two types of film to positron (F-18) and negatron (C-14) emitters. Variations in the response of film to increasing film-to-source distance are described, along with the effects of different intensifying screens and mounting tape. Digitization of whole-body autoradiograms (WBARG) in small animals was performed by using a videodensitometry system (videocamera interfaced to a computer). Quantitation was derived from analysis of a series of step-wedge standards that covered the range of radioactivities in the sample. By using a close-up lens on the videocamera, a 2- by 2-cm field is digitized as a 128 X 128 array, each pixel representing 156 X 156 micron. The effect of chlorpromazine (CPZ) on glucose metabolism in mice was studied by giving C-14 2DG followed by CPZ and F-18 FDG in the same animal. Muscle activity decreased and brown-fat activity increased. The high spatial resolution of this technique enables quantification in structures as small as the basal ganglia in mice. The use of dual-nuclide ARG permits each animal to be its own control, which greatly increases the utility of this method.

Chlorpromazine distribution in hamsters and mice bearing transplantable melanoma.

Chlorpromazine (CPZ) distribution was measured in tissues of Syrian golden hamsters bearing Greene melanoma and in BALB/c mice bearing Harding-Passey melanoma. Distribution was evaluated as a function of time (0.5 to 14 days) and as a function of single and multiple doses (up to five) of from 5 to 50 mg CPZ per kg body weight. Routes of administration (i.p., i.v., p.o.) were compared. The physiological behavior of CPZ is of interest as it is used extensively as a tranquilizing drug (Thorazine). Further, since CPZ binds to the pigment melanin, the possibility exists of using CPZ to transport diagnostic or therapeutic agents to melanoma. It was found that, at 2 days postinjection, tumor/tissue concentration ratios exceeded 10 for metabolizing organs, such as liver and 100 for "back-ground" tissues, such as blood and muscle. Absolute concentrations of CPZ in tumor exceeding 100 microgram CPZ per g tumor were obtained with both single and multiple doses. This selective high concentration in tumor would make CPZ an ideal vehicle for the transport of boron to tumor for use in neutron capture therapy via the 10B(n, alpha)7Li reaction.

Whole-body localization of 14C-tocopheryl acetate in the rat following oral administration.

The whole-body localization of dl-alpha-[3, 4-14C2]-tocopheryl acetate was examined in the rat for intervals ranging between 12-96 hr following administration of a single oral dose. Quantitative evaluation of the macroautoradiograms revealed a rapid removal of 14C-vitamin E equivalents from the blood and gut and their accumulation in body tissues. When the densitometric data were expressed as dpm/mm2, the values for radioactivity uptake by the adrenal cortex were systematically the highest of all tissues examined for all time periods. At 12-96 hr after dosing, high activity was noted in spleen, bone marrow, liver, lymph nodes and fat; moderate activity was observed in myocardium, lung, gastric mucosa, pituitary, blood, hair follicles, Harder's gland and nasal mucosa; low values were found in brain, skeletal muscle and spinal cord. The nearly complete exclusion of radiovitamin E from the brain and its localization to the choroid plexus implied the operation of a blood-brain barrier. Labeled vitamin E uptake in brain, salivary glands and skeletal muscle was essentially linear and increased with time over 12-96 hr. For both early and late sacrifice periods, a differential pattern of 14C-radioactivity uptake was observed with the pituitary, where the pars nervosa exhibited 2.8 times greater activity than the pars distalis. The significance of the findings is discussed in relation to the suggested role of vitamin E in pituitary-adrenal functions, hemopoiesis and oral physiology.