Melatonin and abnormal movements induced by L-dopa in mice.

Melatonin has blocked adventitious movements induced by L-dopa in intact mice. It has reversed the adventitious turning to the right, and it has induced running to the left in mice receiving L-dopa after a lesion in the right caudate nucleus.

Propylbutyldopamine. Mechanism of blood pressure lowering in hypertensive patients.

The effects of propylbutyldopamine ( PBDA ), an analog of dopamine lacking significant vasoconstrictor effects, were examined in seven patients with essential hypertension. Cardiovascular hemodynamics, renal plasma flow, urinary sodium excretion, and the renin-angiotensin-aldosterone system were examined during PBDA infusion both before and after administration of low (8 micrograms/kg) and high (40 micrograms/kg) doses of the dopamine receptor antagonist metoclopramide. Infusion of PBDA at a rate of 20 micrograms/kg/min lowered mean arterial pressure from an average control value of 112 +/- 4 to 94 +/- 3 mm Hg during the last 5 minutes of infusion (p less than 0.01), and increased effective renal plasma flow from 330 +/- 22 ml/min to 591 +/- 46 ml/min (p less than 0.01). Changes in heart rate (+ 16% +/- 5% increase from control values of 77 +/- 3 bpm), urinary sodium excretion (+ 13% +/- 5% increase from control value of 121 +/- 11 muEq/min), plasma renin activity (+ 23% +/- 15% increase over control value of 1.3 +/- 0.3 ng angiotensin I/ml/hr), and plasma aldosterone (+ 26% +/- 12% increase over control value of 17 +/- 6 ng/dl) accompanied PBDA infusion. Pretreatment with metoclopramide at a dose of 8 micrograms/kg prior to PBDA infusion partially blunted the blood pressure reduction produced by PBDA alone (-10% +/- 8% vs -20% +/- 4% compared to control values, p less than 0.1) but had no effect on PBDA -induced increases in renal plasma flow (+ 179% +/- 15% vs + 179% +/- 9% compared to control).(ABSTRACT TRUNCATED AT 250 WORDS)

Structure--activity relationships of n-substituted dopamine and 2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene analogues: behavioral effects in lesioned and reserpinized mice.

N,N-Disubstituted dopamine and 2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene (6,7-ADTN) analogues were synthesized and tested intraperitoneally in mice for dopamine agonism. Compounds inducing asymmetric postures in unilaterally caudectomized mice were further tested in mice treated with reserpine and alpha-methyl-p-tyrosine methyl ester. ED50 values determined for reversal of reserpine-induced catalepsy were used to rank drug potency and correlated with molecular structure. N-n-Propyl N-substituted compounds were more effective than other N,N-dialkyl homologues. Of these, analogues with one alkyl group larger than propyl became inactive or their dopaminomimetic effect was reduced when the propyl was replaced with a larger group. N-Monosubstituted analogues were inactive as dopamine agonists. N-n-P-r-N-n-Bu-6,7-ADTN was six times more potent than N-n-propyl-N-n-pentyl- and N-n-propyl-N-phenethyldopamine but ten times less potent than apomorphine. The availability of an array of structurally related dopamine analogues with dopaminomimetic properties may make it possible to test the hypothesis that there are more than one type of dopamine receptor and that stereotypy and locomotor activity may have different central nervous system loci. Moreover, they may be of potential use in the treatment of parkinsonism and other extrapyramidal disorders.

Cholinergic effects of molecular segments of apomorphine and dopaminergic effects of N,N-dialkylated dopamines.

The hydrochlorides of molecular segments of apomorphine [2-(3',4'-dihydroxybenzyl)-1,2,3,4-tetrahydroisoquinoline, 2-(3'4'-dihydroxybenzyl)piperidine, and 1,2,3,4-tetrahydroisoquinoline with their respective N-methyl and N-n-propyl homologs] and N,N-dialkylated dopamine compounds were synthesized and studied for (1) LD50 in intact mice; (2) stereotypy in intact mice; (3) curving of the body in unilaterally caudectomized mice; (4) rotation in 6-hydroxydopamine-lesioned rats, and (5) activation of adenylate cyclase in homogenates of mouse caudate nuclei. Instead of dopaminergic effects 1-(3',4'-dihydroxybenzyl)-2-methyl-1,2,3,4-tetrahydroisoquinoline and 2-methyl-1,2,3,4-tetrahydroisoquinoline showed cholinergic ones. These effects were blocked in atropine-pretreated animals. Of the N,N-dialkylated dopamine compounds synthesized, the N-n-propyl-N-n-butyldopamine ranked in all tests as the strongest dopamine-receptor agonist and N-methyl-N-n-propyldopamine as the weakest. In contrast, N,N-dimethyldopamine and 1-(3,4-dihydroxyphenylethyl)piperidine showed no dopaminergic effects. The effectiveness of the dopaminergic agonists depended on the length of the N-alkyl substituents suggesting interactions with hydrophobic regions of the receptor site.

Synthesis of [2-11C]5,5-dimethyl-2,4-oxazolidinedione for studies with positron tomography.

We have developed a method for the synthesis of [2-11C]5,5-dimethyl-2,4-oxazolidinedione ([2-11C]DMO) for use with positron emission tomography to measure regional tissue pH in vivo in man. [2-11C]Dimethyl carbonate (DMC) was prepared from [C-11]phosgene and excess of sodium methoxide in methanol containing dimethyl carbonate as added carrier. The [2-11C]DMC solution was then reacted with 2-hydroxyisobutyramide at 150 degrees C for 10 min to yield, after HPLC separation, [2-11C]DMO with a radiochemical yield of 20-56%. Chemical yields were 78-92%, and specific activity ranged as high as 830 mCi/millimol.

[13N]cisplatin PET to assess pharmacokinetics of intra-arterial versus intravenous chemotherapy for malignant brain tumors.

The biodistribution, blood clearance, and in vivo transformation of cisplatin (cisdiaminedichloroplatinum, DDP) were studied in rats using 13N-labeled and unlabeled DDP. Following the i.v. injection of [13N]DDP, virtually no 13N activity was detected in brain tissue, and no measurable amount of the 13N label was displaced from [13N]DDP. Based on these results, [13N]DDP/positron emission tomographic (PET) scans were performed in two glioblastoma patients undergoing Phase II intra-arterial (i.a.) DDP chemotherapy: [13N]DDP was infused i.v. over 13-15 min, during which time serial PET scans were obtained. One hour later, [13N]DDP mixed with cold DDP (100 mg/m2 therapeutic dose) was infused at the same rate i.a., and a second sequence of PET scans was acquired. The pharmacologic advantage of i.a. administration was calculated as the ratio of integrated tumor/brain count ratios for the i.a. and i.v. studies. Our preliminary results demonstrate the feasibility of quantifying the pharmacologic advantage of i.a. DDP chemotherapy in individual brain tumor patients using [13N]DDP and PET.

Imaging of brain tumor proliferative activity with iodine-131-iododeoxyuridine.

METHODS: Iodine-131-iododeoxyuridine (IUdR) uptake and retention was imaged with SPECT at 2 and 24 hr after administering a 10-mCi dose to six patients with primary brain tumors. The SPECT images were directly compared to gadolinium contrast-enhanced MR images as well as to [18F]fluorodeoxyglucose (FDG) PET scans and 201Tl SPECT scans. RESULTS: Localized uptake and retention of IUdR-derived radioactivity was observed in five of six patients. The plasma half-life of [131I]IUdR was short (1.6 min) in comparison to the half-life of total plasma radioactivity (6.4 hr). The pattern of [131I]IUdR-derived radioactivity was markedly different in the 2-hr compared to 24-hr images. Radioactivity was localized along the periphery of the tumor and extended beyond the margin of tumor identified by contrast enhancement on MRI. The estimated levels of tumor radioactivity at 24 hr, based on semiquantitative phantom studies, ranged between < 0.1 and 0.2 microCi/cc (< 0.001% and 0.002% dose/cc); brain levels were not measurable. CONCLUSIONS: Iodine-131-IUdR SPECT imaging of brain tumor proliferation has low (marginal) sensitivity due to low count rates and can detect only the most active regions of tumor growth. Imaging at 24 hr represents a washout strategy to reduce 131I-labeled metabolites contributing to background activity in the tumors, and is more likely to show the pattern of [131I]IUdR-DNA incorporation and thereby increase image specificity. Iodine-123-IUdR SPECT imaging at 12 hr and the use of [124I]IUdR and PET will improve count acquisition and image quality.

Inhibition of glutamate-aspartate transaminase by beta-methylene-DL-aspartate.

beta-Methylene-DL-aspartate, a new beta, gamma-unsaturated amino acid, is an irreversible inhibitor of soluble pig heart glutamate-aspartate transaminase (Ki approximately 3 mM with respect to the L-form; limiting rate constant for inactivation approximately 0.4 min-1). The new amino acid is the most specific inhibitor of glutamate-aspartate transaminase thus far studied. It does not inactivate pig heart glutamate-alanine transaminase, soluble rat kidney glutamine transaminase K, gamma-aminobutyrate transaminase (from Pseudomonas fluorescens), glutamate decarboxylase (Escherichia coli), snake venom L-amino acid oxidase, or hog kidney D-amino acid oxidase. In addition, the following enzymes were not inhibited by beta-methylene-DL-aspartate in rat tissue homogenates: gamma-aminobutyrate transaminase (brain), tyrosine transaminase (liver), glutamine transaminase L (liver), asparagine, transaminase (liver), ornithine transaminase (liver) or branch-chain transaminase(s) (kidney). Intraperitoneal injection of beta-methylene-DL-aspartate into mice decreased kidney and liver glutamate-aspartate transaminase activities but had no effect on liver glutamate-alanine transaminase activity.

Evaluation of peripheral dopamine receptor and alpha-adrenoceptor blocking activity of sulpiride.

The effect of sulpiride, a dopamine receptor antagonist, on peripheral alpha-adrenoceptors and dopamine receptors was determined. Positive chronotropic responses to cardioaccelerator nerve stimulation in pentobarbital-anesthetized dogs were potentiated by 1.0 and 2.0 mg/kg but not by 0.5 mg/kg intravenous sulpiride. This effect persisted after neuronal uptake blockade with desipramine, but was prevented by alpha 2-adrenoceptor blockade with yohimbine. Positive chronotropic effects of intravenous norepinephrine were unchanged by sulpiride, suggesting that sympathetic nerve function was facilitated via a presynaptic mechanism. Since yohimbine prevented the facilitatory action of sulpiride, an analysis of the ability of sulpiride to antagonize alpha-adrenoceptors was made. The reduction in stimulus-induced tachycardia caused by the alpha 2-adrenoceptor agonist tramazoline was significantly antagonized by sulpiride. Furthermore, while sulpiride did not antagonize the pressor effect of the alpha-adrenoceptor agonist phenylephrine, it significantly attenuated the increases in blood pressure produced by tramazoline. The alpha 2-adrenoceptor blocking action of sulpiride lasted for approximately 15 min at 1.0 mg/kg adn for 90 min at 2.0 mg/kg. In additional experiments, it was determined that the dopamine receptor-mediated bradycardic and hypotensive effects of N,N-di-n-propyldopamine were antagonized for at least 2 h by 0.1, 0.5, and 1.0 mg/kg sulpiride. These studies establish that peripheral neuronal and vascular dopamine receptors may be antagonized by sulpiride without affecting alpha-adrenergic mechanisms. However, at doses of 1.0 mg/kg and higher, sulpiride facilitates sympathetic nerve function via a preferential antagonism of alpha 2-adrenoceptors.

Presynaptic dopamine receptors and alpha-adrenoceptors as mediators of the bradycardic action of N-n-propyl-N-n-butyl dopamine.

The role of presynaptic receptors in the bradycardic action of N-n-butyl dopamine (PBDA) was investigated. A biphasic effect on blood pressure and a decrease in heart rate were seen upon intravenous administration of PBDA to pentobarbital-anesthetized dogs. The bradycardia produced by PBDA was unaffected by bilateral vagotomy; however, it was abolished by cardiac sympathetic denervation. When PBDA was readministered following restoration of the cardiac sympathetic nerve activity by electrical stimulation, at frequency of 1 Hz, decrease in heart rate was again observed. The cardioinhibitory action of PBDA was completely abolished by sulpiride, whereas phentolamine and yohimbine caused only partial attenuation, suggesting the involvement of both presynaptic alpha-adrenoceptors as well as dopamine receptors in the bradycardiac action of PBDA. Additional experiments were performed to study the influence of stimulus frequency on the cardioinhibition produced by PBDA. Administration of PBDA to animals with different levels of cardiac sympathetic nerve activity (0.25-2 Hz) resulted in decreases in heart rate. However, yohimbine antagonized this action of PBDA only at the two higher frequencies of cardiac nerve stimulation. Sulpiride completely abolished the bradycardia observed at all the different frequencies of cardiac nerve stimulation. These results demonstrate that activation of presynaptic receptors on cardiac sympathetic nerves can result in a decrease in heart rate. PBDA causes bradycardia via an action on presynaptic dopamine receptors when the cardiac sympathetic nerve activity is low, while both presynaptic dopamine receptors as well as alpha-adrenoceptors are involved in the decrease in heart rate produced by this compound at higher levels of sympathetic nerve activity.

Synthesis of [2-11C]5,5-dimethyl-2,4-oxazolidinedione with and without added dimethyl carbonate as a carrier for studies with positron tomography.

Two methods were developed for the synthesis of [2-11C]5,5-dimethyl-2,4-oxazolidinedione ([2-11C]DMO) for use with positron emission tomography to measure regional cerebral tissue pH in vivo in man. In both methods, A and B, [2-11C]dimethyl carbonate (DMC) was prepared from [11C]phosgene and excess of sodium methoxide in methanol containing 2-hydroxyisobutyramide (HIBA). In method A, an excess of DMC was used as a carrier, while in method B none was used. In both methods, the [2-11C]DMC solution was then heated for 10 min at 150 degrees +/- 2 degrees C causing the reaction of [2-11C]DMC with HIBA to yield [2-11C]DMO with a radiochemical purity of greater than 99%. Method A gave significantly higher radioactive yields, a pure organic product, but lower specific activities. Flash chromatography was used for the separation and purification of [2-11C]DMO prepared by method B.

Dopaminergic antagonists: effects of 1,2,3,4-tetrahydroisoquinoline and its N-methyl and N-propyl homologs on apomorphine- and L-dopa-induced behavioral effects in rodents.

N-Methyl-1,2,3,4-tetrahydroisoquinoline (MTIQ) antagonized apomorphine (APO)-induced stereotypy in a dose-related manner when injected i.p. in rats and attenuated L-dopa-induced hyperactivity in mice. 1,2,3,4-Tetrahydroisoquinoline (TIQ) and its homolog, N-n-propyl-, also blocked APO-induced stereotypy when given similarly. No significant difference was found between the amounts of radioactivity in the brain homogenates of MTIQ- and saline-pretreated rats after injection with [3H] APO. This suggested that MTIQ did not antagonize the behavioral effects of APO by blocking its entry into the brain. Mice fed ad libitum for 90 days with Purina Chow mixed with TIQ (5.0 mg/g) displayed behavioral supersensitivity in comparison with controls when injected with L-dopa (0.40 g/kg i.p.) after pretreatment with carbidopa. This was parallelled by a significant increase of dopamine-related adenylate cyclase activity measured in homogenates of caudate nuclei. The similarity between the behavioral effects induced by some neuroleptics and those observed with TIQ and its homologs suggests that the latter may be a new class of short-acting neuroleptics.

Intravenous self-administration of dopamine receptor agonists by rhesus monkeys.

Pharmacological studies have provided important information relevant to the behavioral role of central nervous system (CNS) dopamine (DA) as well as the existence of multiple DA receptors in the CNS. In the present experiment, the i.v. self-administration of several compounds that are direct DA receptor agonists was evaluated in rhesus monkeys. Apomorphine, piribedil, propylbutyldopamine and bromocriptine were self-administered by at least half of the animals tested, whereas SKF 38393 failed to maintain self-administration. Each of the compounds that was self-administered is an agonist at the DA2 receptor that has been demonstrated in the periphery, whereas SKF 38393 is principally a DA1 agonist. The results suggest that a DA receptor that is similar to the DA2 receptor is involved in this behavioral effect. In addition, the results are consistent with the hypothesis that CNS DA is involved in the reinforcing properties of psychomotor stimulants.

Monoamine oxidase and cerebral uptake of dopaminergic drugs.

The brain uptake of amines that do not enter the brain or enter it poorly was promoted by noncompetitive inhibitors of monoamine oxidase, as shown by behavioral and chemical criteria. Mice pretreated with water or enzyme inhibitors other than those mentioned were placid after receiving dopamine (3,4-dihydroxyphenethylamine). Mice pretreated with monoamine oxidase inhibitors (nialamide or iproniazid) showed upon treatment with dopamine the brisk motor responses characteristic of treatment with its precursor, L-dopa (3,4-dihydroxyphenylalanine). After receiving dopamine, intact nialamide-pretreated mice showed marked increases of brain dopamine, in contrast to water-pretreated test mice or water-treated controls. In unilaterally caudectomized, nialamide-pretreated mice, dopamine induced marked lateral curving of the body toward the lesion followed by running in that direction. Noradrenaline or adrenaline induced curving in caudectomized mice, whereas intact ones remained placid.These catecholamines are bound and inactivated by monoamine oxidase. The cerebral uptakes of chemicals that are bound but not inactivated by monoamine oxidase were thereafter tested. Nialamide induced increased behavioral responses to apomorphine and to N-propyl noraporphine, increased cerebral concentrations of both, and a deep coloration of the brain from methylene blue (bound by monoamine oxidase) but not Evans blue (bound by albumin). Even large doses of nialamide, however, failed to affect the behavioral responses to oxotremorine, which has cholinergic rather than adrenergic or dopaminergic properties. Mitochondrial monoamine oxidase seems therefore to play a specific regulatory role in the transport of substances that it binds, either to inactivate or to release them.

In vivo measurement of regional brain tissue pH using positron emission tomography.

Carbon-11-labeled dimethyloxazolidinedione ([11C]DMO) was injected intravenously into human subjects, and serial positron emission tomographic (PET) scans were obtained until brain-blood equilibration was achieved or could be accurately predicted from dynamic PET and 11C blood data. Knowledge of regional brain-blood partition coefficients for DMO, together with measurements of arterial blood hematocrit and pH, permitted the calculation of regional brain tissue and tumor pH (rpH). [11C]DMO PET rpH values were similar to rpH values derived from quantitative autoradiographic measurements of [14C]DMO concentrations in rat brain slices.

Interaction of soluble pig heart glutamate-aspartate transaminase with various beta,gamma-unsaturated amino acids.

beta-Ethylidene-DL-aspartate (beta EA) and beta-methylene-DL-glutamate (beta MG) were synthesized and tested as potential suicide inhibitors of soluble pig heart glutamate-aspartate transaminase (sGAT). beta MG was found to be a) a substrate with a very low turnover number relative to glutamate and b) a competitive inhibitor with respect to aspartate (albeit with a large binding constant). At high concentrations beta MG inactivated the enzyme but only very slowly. beta EA was also found to be a substrate with a very low turnover number; it did not inactivate the enzyme (1 hr, 25 degrees C) even at a high concentration. However, beta EA was found to bind to the enzyme with an affinity comparable to that of aspartate and glutamate. beta-Methylene-DL-aspartate (beta MA) has been shown to rapidly inactivate glutamate-aspartate transaminase. Therefore, it appears that glutamate-aspartate transaminase can bind analogues of aspartate with alkene groups in the beta position. The conjugated carbonyl groups of beta MA and beta EA will enhance Michael addition in comparison with that expected for vinylglycine. On the other hand, the presence of the methyl groups should reduce the electrophilicity of the double bond of beta EA compared to beta MA. This deactivation and/or steric hindrance to Michael attack may account for the inability of beta EA to inactivate sGAT. Therefore, it may be possible to design selective suicide inhibitors of glutamate-aspartate++ transaminase with the following structure: HO2CC(= CHX)CH(CO2H)NH2, where X is an electron-withdrawing group. Ideally, X would increase the reactivity of the double bond while affording a minimum of steric hindrance to susceptible enzyme-bound bases.