Putative Dopamine Autoreceptor Agonist Research Articles

Pre- and postsynaptic dopaminergic activities of U-86170F

The biochemical, endocrine, receptor binding, and behavioral effects of the putative dopamine autoreceptor agonist, U-86170F, were evaluated in various in vivo and in vitro models. U-86170F and apomorphine were shown to cause a significant reversal of the effects of gamma-butyrolactone (GBL) on dopamine accumulation in mouse striata. In contrast to apomorphine, U-86170F had a ceiling effect on the extent of the reversal of GBL effects (55%), whereas apomorphine had an 82% reversal. The effect on striatal homovanillic acid (HVA) levels was also monitored, and both compounds exerted a similar and significant reduction in striatal HVA. A comparison was made between the effects of intraperitoneal (i.p.) and oral administration of U-86170F in the alpha-methyl-p-tyrosine (alpha-MPT)/prolactin model in rats. When administered by the i.p. route, U-86170F suppressed the effects of alpha-MPT on prolactin level increase, having an ED50 of about 0.03 mg/kg, and when administered by the oral route, its ED50 was approximately 0.1 mg/kg. U-86170F has been shown to be a potent dopamine autoreceptor agonist in the GBL, prolactin, and HVA models, with an effective i.p. dose of approximately 0.03 mg/kg. When evaluated for postsynaptic dopaminergic activity in the reserpinized mouse model, and compared to apomorphine, U-86170F was found to increase locomotor activity, but its maximum effect was only 65% of that attained with apomorphine. Higher doses were needed for postsynaptic effects. In receptor binding studies using cloned D2 receptor preparations, U-86170F was found to exhibit agonist binding properties similar to dopamine as demonstrated by their inhibition of 3H-raclopride binding.(ABSTRACT TRUNCATED AT 250 WORDS)

Occurrence of yawning and decrease of prolactin levels via stimulation of dopamine D2-receptors after administration of SND 919 in rats

SND 919 [S)-2-amino-4,5,6,7-tetrahydro-6-propylamino-benzothiazole) is expected to have a potent and selective dopamine D2-receptor agonistic activity. From this information, the present study was performed to investigate effects of SND 919 on yawning behavior and prolactin secretion in rats. Subcutaneous injections of SND 919 (25-500 micrograms/kg, s.c.) elicited yawning responses. Its dose-response curve was bell-shaped with maximal effects at a dose of 100 micrograms/kg. Yawning behavior was also evoked by the putative dopamine autoreceptor agonists, talipexole (6-allyl-2-amino-5,6,7,8-tetrahydro-4H-thiazolo [4,5-d]azepine) (B-HT 920) (5-100 micrograms/kg, s.c.) and (+)-3-PPP ((+)-3-(3-hydroxyphenyl)-N-n-propylpiperidine) (5-15 mg/kg, s.c.). The yawning induced by SND 919 (100 micrograms/kg, s.c.) as well as talipexole (25 micrograms/kg, s.c.) was inhibited by pretreatment with dopamine D2-receptor antagonists such as spiperone (0.5 mg/kg, i.p.) and YM-09151-2 (cis-N-(1-benzyl-2-methylpyrrolidin-3-yl)-5-chloro-2-methoxy-4-met hylamino- benzamide) (0.1 mg/kg, i.p.), or the muscarinic receptor antagonist, scopolamine (0.5 mg/kg, i.p.). However, the yawning was not affected by the dopamine D1-receptor antagonist, SCH 23390 (R(+)-8-chloro-2,3,4,5-tetrahydro-3-methyl-5-phenyl-1H-3-benzazepine-7-o l) (0.5 mg/kg, i.p.). Stereotypy such as licking and biting was not observed following the administration of SND 919, talipexole and (+)-3-PPP. Administration of SND 919, talipexole or (+)-3-PPP in respective yawn-inducing doses caused a reduction in both the basal prolactin levels and the alpha-methyl-p-tyrosine-induced hyperprolactinemia.(ABSTRACT TRUNCATED AT 250 WORDS)

The dopamine autoreceptor agonist, (±)- Trans-1,3,4,4a5, 10b-hexahydro-4-propyl-2h [1]benzopyrano [3,4-b] pyridin-9-ol hydrochloride (CGS 15855A), modulates striatal dopamine metabolism and prolactin release

The effects of apomorphine and the putative dopamine autoreceptor agonist, CGS 15855A, were evaluated in several functional assays that are modulated by pre- or post-synaptic D 2 receptors. These included release of prolactin in vivo and in vitro from cultured lactotrophs; levels of dihydroxyphenylacetic acid (DOPAC) in the striatum; levels of acetylcholine (ACh); in the striatum and concentrations of cyclic guanosine monophosphate (cyclic GMP) in the cerebellum. The secretion of prolactin was inhibited by CGS 15855A in vitro and in vivo and which also decreased the levels of DOPAC in the striatum at doses 5–25 times less than those required to increase ACh in the striatum and levels of cGMP in the cerebellum. In contrast, apomorphine possessed a dose-ratio between 1.5 and 8.6 for these assay systems. These data suggest that CGS 15855A is a selective dopamine autoreceptor agonist which preferentially stimulates D 2 receptors on lactotrophs and dopaminergic neurons as compared to D 2 receptors on cholinergic interneurons in the striatum.

Further in vitro and in vivo studies with the putative presynaptic dopamine agonist N,N-dipropyl-7-hydroxy-2-aminotetralin.

The in vitro binding of the putative dopamine autoreceptor agonist [3H]DP-7-ATN to rat striatal membrane homogenates was investigated. The maximum number of binding sites Bmax was 497.5 +/- 50.2 fmol/mg protein and the affinity constant KD was 8.3 +/- 1.5 nM using 10 microM (+) butaclamol to define non-specific binding. Lesion of the left medium forebrain bundle by 6-hydroxydopamine resulted in an almost complete loss of dopamine in the striatum but did not affect the binding of [3H]DP-7-ATN. The binding of [3H]DP-7-ATN to the homogenates of the dopaminergic cell bodies in the substantia nigra revealed a Bmax of 542.4 +/- 40.1 fmol/mg protein and a KD of 11.1 +/- 1.3 nM. The pharmacological profile of the binding was characterized as being to D-2 receptors. No direct in vitro evidence could be found for a selective binding to DA autoreceptors. The dopamine uptake inhibitor GBR 12909 interacted in a noncompetitive manner with the in vitro binding of [3H]DP-7-ATN and the latter compounds uptake into isolated synaptosomes was not through the specific dopamine uptake system but rather through diffusion. GBR 12909 failed to reveal any agonistic or antagonistic activity in the GBL model but was able to antagonize the hypomotility in rats induced by 0.25 mg/kg DP-7-ATN. The inhibitory effect of DP-7-ATN on DA release was also demonstrated using in vivo brain dialysis in conscious rats. Based on the above results, the possibility is discussed that the release regulating DA autoreceptors, which might be coupled to the reuptake complex, and the DA biosynthesis regulating autoreceptors, are different entities.

Conditioning of behavioural signs produced by nomifensine and by B-HT 920 in rats.

Conditioning of behavioural effects produced by two drugs acting differently upon dopaminergic neurotransmission was studied. Nomifensine and the putative dopamine autoreceptor agonist B-HT 920 produce contrasting effects on motility, namely increases in locomotor activity and stereotypies as compared to hypokinesia and ptosis. The administration of each of these drugs (US) was repeatedly associated with well-defined environmental stimuli (CS): a wire cage associated with an auditory and on olfactory stimulus. The rats were conditioned for 7 days with 20 mg/kg nomifensine IP each day. After conditioning, the rats were treated with the solvent alone in presence of the CS. Not only did sniffing and licking occur, but also gnawing, even though the latter response was not evident after acute administration of the drug or during the conditioning period. Nomifensine (20 mg/kg IP) also acutely decreased the ratio of 3,4-dihydroxyphenylacetic acid/dopamine concentrations (DOPAC/DOPAMINE); this ratio was not altered in the conditioned rats, 60 min after solvent administration in presence of the CS. Rats were conditioned with 0.02 mg/kg IP B-HT 920 daily for 8 days. During the conditioning phase, akinesia and ptosis showed a slight enhancement and a faster onset. After conditioning, when the rats were treated with the solvent alone, the majority of them showed akinesia and/or ptosis during the observation period, in contrast to pseudoconditioned controls. When these rats were conditioned or pseudoconditioned, respectively, with B-HT 920 for further 5 days using 0.02 mg/kg again, treatment with the same dose in presence of the CS produced a significant enhancement and acceleration of these signs in conditioned as compared with pseudo-conditioned control rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Inhibition of K+-stimulated [3H]dopamine and [14C]acetylcholine release by the putative dopamine autoreceptor agonist, B-HT 920

The inhibition of K+-stimulated [3H]dopamine and [14C]acetylcholine release from preloaded rat striatal slices was used to examine the presynaptic selectivity of the putative dopamine autoreceptor agonist, B-HT 920. In the micromolar range, B-HT 920 caused a concentration-dependent inhibition of the release of both labeled neurotransmitters as evoked by 20 mM K+. The effect of B-HT 920 on both [3H]dopamine and [14C]acetylcholine release was completely blocked by (+) butaclamol but not by (-) butaclamol. Sulpiride, a selective D2 antagonist, similarly blocked the inhibitory effect of B-HT 920 on the release of both labeled neurotransmitters indicating both responses were mediated by D2 receptors. (+) Butaclamol alone elevated stimulated [3H]dopamine release suggesting a significant amount of autoreceptor occupancy by endogenously released dopamine. Experiments with tolazoline and the alpha 2 agonist, B-HT 933, did not suggest any involvement of alpha-adrenoceptor activity in the inhibitory effects of B-HT 920 on the release of either transmitter. Inhibition of release was a selective effect of B-HT 920 as the drug was without effect on the K+-stimulated release of [3H]serotonin. The results indicate that in vitro B-HT 920 is active of both pre- and postsynaptic dopamine receptors in contrast to the pattern of effects observed after its in vivo administration.

Involvement of catechol- O-methyl transferase in the metabolism of the putative dopamine autoreceptor agonist 3-PPP(3-(3-hydroxyphenyl)- N- n-propylpiperidine)

Involvement of catechol- O-methyl transferase in the metabolism of the putative dopamine autoreceptor agonist 3-PPP(3-(3-hydroxyphenyl)- N- n-propylpiperidine)

The putative dopamine autoreceptor agonist B-HT 920 decreases nigral dopamine cell firing rate and prolactin release in rat

Recent behavioural and biochemical investigations have suggested that the alpha-2 receptor agonist B-HT 920 is also a centrally acting dopamine (DA) agonist with a selectivity for autoreceptors. It is presently demonstrated that B-HT 920, in contrast to the structurally related alpha-2 agonist B-HT 933, effectively reduces the firing rate of nigral DA neurons both after intravenous and microiontophoretic administration. Furthermore, B-HT 920, but not B-HT 933, decreases plasma levels of prolactin in reserpine pretreated rats. The electrophysiological as well as the neuroendocrine effects of the drug were antagonised by DA antagonists but not by alpha-2 receptor antagonists. The data support the contention that B-HT 920 acts as an agonist at central DA autoreceptors. Furthermore, they reinforce the hypothesis that lactotroph DA receptors are more similar to DA autoreceptors than to postsynaptic DA receptors in the brain.

Is inhibition of striatal synaptosomal tyrosine hydroxylation by dopamine agonists a measure of dopamine autoreceptor function?

Rat striatal synaptosomal tyrosine hydroxylation was inhibited dose- and pH dependently by a number of dopamine agonists. The catecholic agonists apomorphine and (-)N-n-propylnorapomorphine inhibited synaptosomal tyrosine hydroxylase completely, with IC50 values of around 0.3 mumol/l at pH 6.6. The noncatechol agonists pergolide and bromocriptine and the putative dopamine autoreceptor agonists 3-PPP(-), 3-PPP(+), HW-165 and B-HT 920 produced only partial inhibition of synaptosomal tyrosine hydroxylation at high concentrations. Comparison of the inhibition of synaptosomal and soluble tyrosine hydroxylase indicated that the inhibition produced by apomorphine could be ascribed to a direct effect on the enzyme, whereas this was not the case for the noncatechol agonists. The inhibition produced by pergolide and 3-PPP(-) was not antagonised by either dopamine receptor or alpha-adrenoceptor antagonists. The present results have been compared with results reported in the literature for inhibition of synaptosomal tyrosine hydroxylation and for two other tests of dopamine autoreceptor agonist activity (inhibition of dopamine release from striatal slices in vitro, and inhibition of the gamma-butyrolactone induced increase in dopamine synthesis in vivo). It is concluded that inhibition of striatal synaptosomal tyrosine hydroxylation by dopamine agonists does not fulfil the criteria required for it to be considered as a useful measure of dopamine autoreceptor function.

Dopamine synthesis in synaptosomes: relation of autoreceptor functioning to pH, membrane depolarization, and intrasynaptosomal dopamine content.

Factors affecting dopamine (DA) synthesis in rat striatal synaptosomes were examined by measuring the conversion of [3H]tyrosine (Tyr) to [3H]DA. Any [3H]DA that was synthesized was extracted into a toluene-based scintillation cocktail and quantitated by liquid scintillation spectrometry. The extraction was facilitated using di-(2-ethylhexyl) phosphoric acid (DEHP), a liquid cation exchanger. DA, apomorphine, and other DA agonists were much less potent inhibitors of DA synthesis in striatal synaptosomes at pH 6.2 than at pH 7.2. 3-(3-Hydroxyphenyl)-N-n-propylpiperidine (3-PPP), a putative DA autoreceptor agonist, was inactive at pH 6.2. However, at pH 7.2, 3-PPP did inhibit DA synthesis. This inhibition was reversed by sulpiride, a DA receptor antagonist, but not by benztropine, a DA uptake blocker, suggesting that 3-PPP inhibits DA synthesis by stimulating the DA autoreceptor. DA release from synaptosomes was much greater at pH 6.2 than at pH 7.2, most probably because the synaptosomal membrane appears to be depolarized at pH 6.2, as measured by the accumulation of [3H]tetraphenylphosphonium ions. Since tyrosine hydroxylase is inhibited by DA, this finding suggested that low assay buffer pH (i.e., pH 6.2) might interfere with the ability of 3-PPP and other DA agonists to inhibit DA synthesis, by promoting DA release. Likewise, reserpine and tetrabenazine, compounds which disrupt vesicular DA storage, were much less effective inhibitors of DA synthesis at pH 6.2 (high basal DA release). Moreover, D-amphetamine and high buffer potassium concentrations, treatments which promote DA release, also interfered with the ability of 3-PPP to inhibit DA synthesis. Thus, modulation of the release of DA in equilibrium with tyrosine hydroxylase may be a mechanism by which the DA autoreceptor regulates DA synthesis.

Effects of S (+)-3-phenethyl-PP, a putative dopamine autoreceptor agonist with greater autoreceptor selectivity than 3-PPP enantiomers

The experiments concerned the pharmacology of the enantiomers of the phenethyl-analogue (3-phenethyl-PP) of the putative dopamine (DA) autoreceptor agonist 3-PPP. In contrast to the almost equipotency of 3-PPP enantiomers, the phenethyl enantiomers showed marked stereoselectivity. S(+)-3-Phenethyl-PP had 25 times higher affinity to D-2 DA receptors in vitro than the R(−)-enantiomer. In vivo a similar potency difference was seen for the inhibition of motility, induction of circling behaviour in 6-OHDA-lesioned rats and emetic effect in dogs. None of the enantiomers induced stereotypy and hypermotility in normal rats or in rats pretreated with reserpine and α-methyl-p-tyrosine. In test models for antidopaminergic activity only slight activity of either enantiomer was observed. The S(+)-enantiomer had no antagonistic effect against apomorphine- and amphetamine-induced stereotypies, no cataleptogenic activity and only partially antagonized amphetamine-induced hypermotility. Apomorphine-induced emesis was weakly antagonized. The results indicate a greater and higher selectivity of S(+)-3-phenethyl-PP for DA receptors mediating sedation compared with 3-PPP enantiomers which previously have been shown to exert significant effects on postsynaptic DA receptors. Thus S(+)-3-phenethyl-PP may be a more selective model compound for the differential study of effects elicited by stimulation of pre- and postsynaptic DA receptors.

Interaction of the component enantiomers of the putative dopamine autoreceptor agonist, TL-99 (6,7-dihydroxy-2-dimethylamino tetralin) with dopaminergic systems in mammalian brain and teleost retina.

The enantiomers of the putative dopamine autoreceptor agonist, TL-99 (6,7-dihydroxy-2-dimethylaminotetralin) were examined in a number of in vivo and in vitro test paradigms to further examine the reported autoreceptor selectivity of this compound. The (+)-isomer of the aminotetralin was more active as a dopamine agonist than either the racemate or the (-)-enantiomer. In addition to this dopaminergic activity, TL-99 was found to be a potent alpha 2-adrenoceptor agonist, this activity being more prominent in the (+)-isomer. The (-)-isomer, however, was a weak alpha 2/DA receptor agonist and unlike the (+)-enantiomer was devoid of activity in the D-1-selective carp retina adenylate cyclase assay. Pharmacological examination of the effects of TL-99 on mouse locomotor activity showed that the effects of the aminotetralin in this dopamine autoreceptor test system were antagonized by either the alpha 2-antagonist, yohimbine or by the dopamine antagonist, sulpiride. TL-99 also produced contralateral turning in 6-OHDA lesioned rats. It is concluded that the apparent dopamine autoreceptor selectivity of TL-99 as assessed by in vivo animal test systems may be due partially to its alpha 2-agonist activity. The sedation and consequent reduction in mouse locomotor activity and in turning in the rat as the dose level is increased undoubtedly occurs via alpha 2-agonist and dopamine autoreceptor activity and cannot be interpreted as selectivity for the dopamine autoreceptor.

Electrophysiological studies on EMD 23 448, an indoly l-3-butylamine, in the rat: A putative selective dop amine autoreceptor agonist

Single unit electrophysiological and microiontophoretic techniques were used to study the effects of a putative selective dopamine autoreceptor agonist, EMD 23 448, an indolyl-3-butylamine on the activity of both dopamine neurons in the substantia nigra and dopamine-sensitive neurons in the caudate nucleus. Intravenous administration of EMD 23 448 inhibited the firing of dopamine neurons in the nigra in a dose-dependent manner. This effect could be readily reversed by the subsequent intravenous administration of the dopamine receptor antagonist, haloperidol. Microiontophoretic application of EMD 23 448 onto these neurons also inhibited their activity and this effect was blocked by iontophoresis of the dopamine receptor antagonist, trifluoperazine. Intravenous administration of the former compound increased the activity of dopamine-sensitive cells in the caudate in the same doses which inhibited dopamine neurons in the nigra. However, this effect appeared to be indirect since the iontophoretic application of EMD 23 448 directly onto caudate neurons had no effect on their firing rate. In addition, the activity of norepinephrine-containing neurons in the locus coruleus and serotonin-containing cells in the dorsal raphé nucleus was not changed by direct microiontophoretic application of this compound. In contrast, EMD 23 448 selectively blocked the acetylcholine-induced activation of pyramidal neurons in the hippocampus. These findings provide further evidence that with respect to central catecholamine neurons EMD 23 448 may be a selective agonist for dopamine autoreceptors.

Avoidance and ICSS behavioral models dissociate TL-99 and 3-PPP from dopamine receptor antagonists

The behavioral effects of the putative dopamine autoreceptor agonists, TL-99 and 3-PPP, were explored in animal procedures that reveal highly characteristic effects of neuroleptics currently in clinical use. Sidman avoidance responding in rats was not altered appreciably by doses up to 10 mg/kg TL-99 or 30 mg/kg 3-PPP. Higher doses of TL-99 attenuated Sidman avoidance performance in squirrel monkeys, although 3-PPP had no effect. Lever pressing for intracranial self-stimulation (ICSS) was attenuated in a dose-related fashion by TL-99 and 3-PPP, with relatively shallow dose-response relationships. A low dose of haloperidol (0.03 mg/kg) partly reversed the effects of 3-PPP (3 mg/kg) on lever pressing ICSS, but not those of TL-99 (3 mg/kg). Yohimbine (3 mg/kg) failed to alter the effects of TL-99 at a dose that abolished the suppresant effect of clonidine on ICSS. Analysis of within-session ICSS response decrement patterns indicated that TL-99 reduced ICSS to a greater extent towards the end of the session than during the first 5 min. No such within-session trend was produced by 3-PPP, suggesting that 3-PPP attenuates ICSS by virtue of a performance deficit. Similar conclusions were reached using a shuttlebox task that involved self-regulation of ICSS duration by rats. Therefore, the clinical profile of neuroleptics is unlikely to be mimicked precisely by 3-PPP or TL-99. Clinical trials of DA autoreceptor agonists for antipsychotic efficacy will indicate whether or not avoidance and ICSS behaviors are relevant to the detection of the intrinsic antipsychotic activity of drugs.

Assessment of the chloroethyl analog of 3-PPP (N-n-propyl-3-(3-hydroxyphenyl)-piperidine), 3-PPP-Cl as an irreversible ligand for central dopaminergic recognition sites

NCA, the chloro analog of the potent dopamine agonist NPA is an irreversible ligand at dopamine receptors in mammalian brain. The chloroethyl analog of the recently described putative dopamine autoreceptor agonist 3-PPP, 3-PPP-Cl, was evaluated for its potential use as an irreversible autoreceptor ligand. N-Chloroethylation of 3-PPP reduced the intrinsic affinity of the agonist seven-fold and, consequently, in contrast to NCA, it was found that 3-PPP-Cl was not a good irreversible ligand at dopamine receptors.

Effect of two dopamine agonists, TL-99 and 3-PPP, on prolactin secretion in the rat

TL-99 and 3-PPP, two putative dopamine autoreceptor agonists, effectively suppressed serum prolactin (PRL) concentrations in rats treated with α-methyltyrosine. In addition, the secretion of PRL from anterior pituitary tissue under in vitro conditions was significantly inhibited by TL-99 and 3-PPP. These findings are supportive of the view that TL-99 and 3-PPP exert a dopamine agonist action at dopamine receptors in the anterior pituitary gland.

In vitro effect of the racemic mixture and the (-)enantiomer of N-n-propyl-3-(3-hydroxyphenyl)-piperidine (3-PPP) on postsynaptic dopamine receptors and on a presynaptic dopamine autoreceptor.

The racemic mixture and the (-)enantiomer of the putative dopamine autoreceptor agonist 3-PPP were investigated in vitro using dopamine-sensitive adenylate cyclase in homogenates of rat striatum as a model for a postsynaptic D1-receptor type and inhibition of electrically-evoked tritium overflow from rat striatal slices preincubated with [3H]choline and [3H]dopamine as a model for a postsynaptic D2- and a presynaptic dopamine autoreceptor type, respectively. In contrast, to apomorphine, neither the racemic mixture nor the (-)enantiomer exerted any effect, suggesting agonistic properties in all three receptor models. However, both (+/-)3-PPP and (-)3-PPP were weak antagonists at postsynaptic D1- and D2-receptors. The results of the present investigation suggest that the in vivo effects of 3-PPP are either the result of metabolic activation or that this drug activates an other dopamine autoreceptor type, pharmacologically different from that one modulating dopamine release.

Postynaptic dopamine agonist properties of TL-99 are revealed by yohimbine co-treatment

The claim that TL-99 (6,7-dihydroxy-2-dimethylaminotetralin hydrobromide) is a selective dopamine autoreceptor agonist relies partly upon indirect behavioral evidence, particularly the absence of stereotyped behavior in treated rats. The possibility was examined that concurrent α 2-adrenoceptor agonist properties of TL-99 could have masked postsynaptic dopamine agonist activity. Co-administration of yohimbine or piperoxan with a high dose of TL-99 (30 mg/kg) dramatically increased motor activity in reserpinized rats, whereas each drug by itself had no effect. Contralateral rotational behavior in 6-hydroxydopamine-lesioned rats resulted from combined treatment with yohimbine and a high dose of TL-99 (30 mg/kg) but appeared to be suppressed by concurrent flaccidity if TL-99 was given by itself. Yohimbine failed to alter the effects of 3-PPP (N-n-propyl-3-(3-hydroxyphenyl)-piperidine), another putative dopamine autoreceptor agonist, in either model of postsynaptic dopamine agonism. It is concluded that a concurrent behaviorally depressant action of TL-99, possibly α 2-agonism, masks the stimulation of postsynaptic dopamine receptors by high doses of TL-99.

Effect of 3‐PPP, a putative dopamine autoreceptor agonist, on [3H]ligand binding to dopamine receptors of calf and rat striatum

Abstract3‐(3‐Hydroxyphenyl)‐N‐n‐propyliperidine (3‐PPP) is most effective in inhibiting [3H]apomorphine binding in rat striatal membranes, with Ki values of 63 nM. 3‐PPP was six to 27 times less effective when it competed with the binding of [3H]dopamine or [3H]spiperone in calf and rat striatal membranes. At concentrations up to 10 μM, 3‐PPP failed to substitute for dopamine in the activation of adenylate cyclase in rat striatal membranes. 3‐PPP at 4.8‐5 μM caused 50% inhibition of catecholamine uptake in synaptosomes of corpus striatum and hypothalamus, therefore appearing to be a relatively weak uptake inhibitor. The higher affinity of 3‐PPP for [3H]apomorphine binding sites is consistent with its binding to a subset of dopamine receptors which are characterized by a high affinity for both the agonist and antagonist of dopamine.

Interaction of the putative dopamine autoreceptor agonists 3-PPP and TL-99 with [ 3H]apomorphine binding sites in rat striatal membranes

Examination of the binding of [ 3H]apomorphine to rat striatal membranes in the presence and absence of the dopamine antagonist, domperidone, confirmed the previously reported presence of two classes of dopamine binding site, those designated D 2 which show a high affinity for both agonist and antagonists and those designated D 3 which have a high affinity for agonists and a low affinity for antagonists. In contrast to the previously reported single high affinity (K D ≅ 1 nM) D 2- and D 3-binding sites, two lower affinity sites (D 2K D = 7–50 nM; D 3 K D = 41 nM) were also observed. Examination of the binding characteristics of the putative dopamine autoreceptor agonists, 3-PPP (N,N-propyl-3-(3-hydroxyphenyl)piperdine) and TL-99 (6,7-dihydroxy-2-dimethylaminotetraline) showed that they, like a number of other dopamine agonists including n-propylnorapomorphine, apomorphine and dopamine showed no preferential affinity for the D 3, presynaptic binding site. It is concluded that the selectivity of dopamine agonists for the autoreceptor cannot be assessed by the in vitro radioligand binding parameters defined by the use of domperidone.