Evaluation of 11C-colchicine for PET imaging of multiple drug resistance.

UNLABELLED: Overexpression of P-glycoprotein (P-gp) can confer multiple drug resistance (MDR) phenotype on cancer cells and tumors by reducing intracellular accumulation of various cytotoxic agents. Early diagnosis of MDR in the clinic will serve to improve the efficacy of chemotherapeutic intervention and the quality of life of patients. In this article we describe use of a positron-emitting MDR tracer, 11C-colchicine (CHC), to evaluate MDR by PET imaging. Unlike existing MDR tracers such as 99mTc-sestamibi, this compound is electroneutral, with biodistribution not affected by perturbations of membrane potential. METHODS: In vitro studies showed that resistance to CHC is correlated to resistance to Taxol (paclitaxel). The results of biodistribution experiments were found to be consistent with previously reported experiments with CHC labeled with other isotopes. On the basis of in vitro experiments with a series of drug-resistant variants of the human neuroblastoma BE (2)-C cell line, a mathematic model of 11C-CHC distribution in tumors was formulated. Dynamic PET 11C-CHC imaging experiments were performed with nude rats xenografted with the BE (2)-C-sensitive and -resistant strains. Each scan was accompanied by a transmissions scan and a static FDG scan. These scans allowed improved image localization. RESULTS: We observed an approximately 2-fold difference between 11C-CHC accumulation in sensitive and resistant tumors. Imaging data were analyzed using the mathematic model, and various parameters characterizing resistance could be identified and estimated. In particular, the parameter r, proportional to the level of resistance of the tumors, was obtained. We showed that the ratio of these r parameters determined from the sensitive and resistant tumors was identical to the ratio of CHC accumulation in the corresponding sensitive and resistant cell lines used for xenografting. CONCLUSION: These in vivo experiments provided additional evidence for the indirect effect of P-gp action on CHC-to-tubulin binding, which in turn determines CHC uptake in tumors. The significance of these findings and future plans is discussed.

Preparation of [11C]carbon dioxide from [11C]cyanide.

[11C]Carbon dioxide was prepared from [11C]cyanide. First, [11C]CH4 was produced by the (p, alpha) reaction on a N2/H2 mixture and was converted to [11C]CN- by reaction with ammonia over platinum at 1273 K. The [11C]CN- was adsorbed on a cobalt(II, III) oxide/ceramic furnace material at room temperature and was subsequently converted to [11C]CO2 by heating the cobalt(II, III) oxide to 948 K over a 10 min period and collecting the [11C]CO2 in a trap at 77 K. Specific activities as high as 600 mCi/mumol at end of bombardment (EOB) were obtained in a 15 microA irradiation for 10 min.

Synthesis of nitrogen-13 labeled alkylamines via amination of organoboranes.

Organoboranes react with nitrogen-13 labeled ammonia to produce alkylamines in moderate yield. When 13N labeled ammonia was bubbled into a tetrahydrofuran solution containing 0.5M tridecylborane, 1-[13N]aminodecane was formed in 25-30 min from the end of bombardment (EOB) in 40-60% overall yield. 1-[13N]aminooctane and 1-[13N]aminohexane were also synthesized from appropriate organoboranes in similar yield.

Carbon-11 labeled dialkylketones: synthesis of 9-[11C]heptadecan-9-one.

9-[11C]heptadecan-9-one was synthesized from di-n-octylthexylborane via cyanidation with K11CN. The rearrangement of the organoborane intermediate followed by alkaline oxidation produced the title compound in 55-60 min from the end of bombardment (EOB) in 50-70% overall yield. The reaction sequence is applicable for the synthesis of various dialkyl ketones.

Use of 11C as a tracer for studying the synthesis of [11C]urea from [11C]cyanide.

The use of reversed-phase liquid chromatography and radiochemical detection with carbon-11 (t1/2 = 20.4 min) as a tracer allowed the study of the preparation of [11C]urea from [11C]cyanide at no-carrier-added concentrations. [11C]cyanate was readily prepared by permanganate oxidation of [11C]cyanide at 75 degrees C. The conversion of NH4O11CN (approximately 0.03 mM) to [11C]urea in the presence of excess ammonium ions (0.28 M) was found to best fit pseudo first order reaction kinetics with a rate constant of 0.065 +/- 0.008 min-1 at 75 degrees C. Heating at higher temperatures (180-200 degrees C) revealed that the conversion of NH4O11CN to [11C]urea occurred in high yield in less than 3 min. The hydrolysis of [11C]cyanate to [11C]carbonate, a possible side reaction, was found to proceed at a rate of 0.010 +/- 0.001 min-1 at 113 degrees C.

Measurement of cerebral blood flow in the rat with intravenous injection of [11C]butanol by external coincidence counting: a repeatable and noninvasive method in the brain.

No method has been reported for measuring CBF, repeatedly and noninvasively, in the rat brain. A new method is described, which is noninvasive to the brain, skull, or cervical large vessels. Two pairs of coincidence detectors were positioned, one over the rat brain and the other at the loop of a catheter inserted into the femoral artery. The coincidence head curve and arterial curve were recorded after intravenous injection of 1-[11C]butanol in 15 rats. CBF was calculated by one-compartment curve fitting ( CBFo ) from 1-min data and with the recirculation corrected height/area method from 3-min data ( CBFh X 3 min) and 5-min data ( CBFh X 5 min). CBFo agreed well with CBFh X 5 min, although a slight overestimation was observed in CBFh X 3 min. The normal CBFo in the normocapnic group (n = 6, paCO2 36.7 +/- 2.3 mm Hg) was 1.76 +/- 0.49 ml/g min (mean +/- SD). A good correlation was observed between CBFo (y) and PaCO2 (x), and the regression line was y = 0. 0629x -0.715 (r = 0.88, p less than 0.0001). We concluded that this method gives the stable blood flow values noninvasively and with a minimum loss of blood (less than 0.28 ml per measurement). Applications of this method include activation studies, studies on the effect of drugs and treatments, and water and oxygen extraction fraction studies using different tracers in the same rat.

Substituent branching in phenethylamine-type hallucinogens: a comparison of 1-[2,5-dimethoxy-4-(2-butyl)phenyl]-2-aminopropane and 1-[2,5-dimethoxy-4-(2-methylpropyl)phenyl]-2-aminopropane.

Two novel hallucinogen analogues related to 1-(2,5-dimethoxy-4-methylphenyl)-2-aminopropane (DOM, STP) were synthesized and evaluated in the two-lever drug discrimination paradigm by using 0.08 mg/kg of LSD as the training drug stimulus. The two compounds differ from each other only with respect to the point of branching in the 4-alkyl group. However, pharmacological evaluation revealed a clear difference in difference in potency and degree of LSD generalization for the two isomers. Branching adjacent to the ring, as in the 4-(2-butyl) analogue, may provide steric interference to the formation of the drug-receptor complex, while branching one methylene unit removed from the ring, as in the 4-(2-methylpropyl) analogue, poses less of a steric problem for the drug-receptor interaction. This is consistent with the idea that formation of a charge-transfer complex between the hallucinogen molecule and the receptor may be one of the features of this drug-receptor interaction.

Syntheses with isotopically labelled carbon. Methyl iodide, formaldehyde and cyanide.

Many of the uniquely labelled synthetic precursors currently employed in the design of sophisticated radiolabelled compounds have their origins in the field of hot atom chemistry. Particularly, the development during the past few years of automated, on-line synthetic procedures which combine the nuclear reaction, hot atom and classical chemistry, and rapid purification methods has allowed the incorporation of useful radionuclides into suitable compounds of chemical and biochemical interest. The application of isotopically labelled methyl iodide, formaldehyde, and cyanide anion as synthetic intermediates in research involving human physiology and nuclear medicine, as well as their contributions to other scientific methodology, is reviewed.