morphinans and bremazocine

Itch/pruritus is the most common side effect associated with spinal administration of morphine given to humans for analgesia. The aim of this study was to investigate the effectiveness of kappa-opioid receptor (KOR) agonists with diverse chemical structures as antipruritics and to elucidate the receptor mechanism underlying the antipruritic effect in monkeys. In particular, previously proposed non-KOR-1 agonists, including nalfurafine [TRK-820, 17-cyclopropylmethyl-3,14 beta-dihydroxy-4,5 alpha-epoxy-6 beta-[N-methyl-trans-3-(3-furyl)acrylamido]morphinan], bremazocine [(+/-)-6-ethyl-1,2,3,4,5,6-hexahydro-3-[(1-hydroxycyclopropy)-methyl]-11,11-dimethyl-2,6-methano-3-benzazocin-8-ol], and GR 89696 [4-[(3,4-dichlorophenyl)acetyl]-3-(1-pyrrolidinylmethyl)-1-piperazinecarboxylic acid methyl ester] were studied in various behavioral assays for measuring itch/scratching, analgesia, and respiratory depression. Systemic administration of nalfurafine (0.1-1 microg/kg), bremazocine (0.1-1 microg/kg), or GR 89696 (0.01-0.1 microg/kg) dose-dependently attenuated intrathecal morphine (0.03 mg)-induced scratching responses without affecting morphine antinociception. The combination of intrathecal morphine with these KOR agonists did not cause sedation. In addition, pretreatment with effective antiscratching doses of nalfurafine, bremazocine, or GR 89696 did not antagonize systemic morphine-induced antinociception and respiratory depression. The dose-addition analysis revealed that there is no subadditivity for nalfurafine in combination with morphine in the antinociceptive effect. Furthermore, the KOR antagonist study revealed that antiscratching effects of both nalfurafine and a prototypical KOR-1 agonist, U-50488H [trans-(+/-)-3,4-dichloro-N-methyl-N-(2-[1-pyrrolidinyl]-cyclohexyl)-benzeneacetamide], could be blocked completely by a selective KOR antagonist, nor-binaltorphimine (3 mg/kg). These findings suggest that the agonist action on KOR mainly contributes to the effectiveness of these atypical KOR agonists as antipruritics, and there is no evidence for KOR subtypes or mu-opioid antagonist action underlying the effects of these KOR agonists. This mechanism-based study further supports the clinical potential of KOR agonists as antipruritics under the context of spinal opioid analgesia.

Topics: Analgesia; Animals; Benzomorphans; Female; Hot Temperature; Injections, Spinal; Macaca mulatta; Male; Morphinans; Morphine; Neurons; Piperazines; Pyrrolidines; Reaction Time; Receptors, Opioid, kappa; Restraint, Physical; Spiro Compounds

In the search for opioid agonists with delayed antagonist actions as potential treatments for substance abuse, the bridged morphinan BU74 (17-cyclopropylmethyl-3-hydroxy-[5beta,7beta,3',5']-pyrrolidino-2'[S]-phenyl-7alpha-methyl-6,14-endoetheno morphinan) (3f) was synthesized. In isolated tissue and [35S]GTPgammaS opioid receptor functional assays BU74 was shown to be a potent long-lasting kappa opioid receptor agonist, delta opioid receptor partial agonist and mu opioid receptor antagonist. In antinociceptive tests in the mouse, BU74 showed high efficacy and potent kappa opioid receptor agonism. When its agonist action had waned BU74 became an antagonist of kappa and mu opioid receptor agonists in the tail flick assay and of delta, kappa and mu opioid receptor agonists in the acetic acid writhing assay. The slow onset, long-duration kappa opioid receptor agonist effects of BU74 suggests that it could be a lead compound for the discovery of a treatment for cocaine abuse.

Topics: Acetic Acid; Analgesics; Animals; Benzomorphans; Binding, Competitive; Bridged-Ring Compounds; Cell Line, Tumor; Guanosine 5'-O-(3-Thiotriphosphate); Guinea Pigs; Hot Temperature; Ileum; In Vitro Techniques; Male; Mice; Mice, Inbred ICR; Morphinans; Narcotic Antagonists; Pain; Pain Measurement; Radioligand Assay; Receptors, Opioid, delta; Receptors, Opioid, kappa; Receptors, Opioid, mu; Sulfur Radioisotopes; Vas Deferens

Cocaine is thought to be addictive because chronic use leads to molecular adaptations within the mesolimbic dopamine (DA) circuitry that affect motivated behavior and emotion. Although the reinforcing effects of cocaine are mediated primarily by blocking DA reuptake into the presynaptic nerve terminal, reciprocal signaling between DA and endogenous opioids has important implications for cocaine dependence. The present study used the opioid antagonist 6 beta-[125iodo]-3,14-dihydroxy-17-cyclopropylmethyl-4,5 alpha-epoxymorphinan ([125I]IOXY) after pretreatment with the site-directed acylating agents 2-(p-ethoxybenzyl)-1-diethylaminoethyl-5-isothiocyanatobenzimid iazole -HCl (mu-selective) and N-phenyl-N-[1-(2-(4-isothiocyanato)-phenethyl)-4-piperidinyl]-p ropana mide-HCl (delta-selective) to examine the effect of cocaine exposure on the distribution and density of kappa2 receptors in autopsy studies of human cocaine fatalities. The selective labeling of the kappa2 receptor subtype was demonstrated by competition binding studies, which gave a pharmacological signature (IOXY >/= (+)-bremazocine >> U50,488 >/= U69,593) distinct from either the kappa1 or kappa3 receptor subtypes. Visualization of [125I]IOXY labeling revealed that kappa2 receptors localize to mesocortical and subcortical limbic areas, including the cingulate, entorhinal, insular, and orbitofrontal cortices and the nucleus accumbens and amygdala. The number of kappa2 receptors in the nucleus accumbens and other limbic brain regions from cocaine fatalities was increased twofold as compared with age-matched and drug-free control subjects. Cocaine overdose victims, who experienced paranoia and marked agitation before death, also had elevated densities of kappa2 receptors in the amygdala. These findings demonstrate for the first time that kappa2 receptor numbers are upregulated by cocaine exposure. The molecular adaptation of kappa2 receptor numbers may play a role in the motivational incentive associated with episodes of binge cocaine use and in the dysphoria that follows abrupt cocaine withdrawal.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Adult; Amygdala; Benzeneacetamides; Benzomorphans; Binding, Competitive; Brain; Brain Chemistry; Caudate Nucleus; Cocaine; Dopamine; Dopamine Uptake Inhibitors; Drug Overdose; Female; Fentanyl; Gyrus Cinguli; Humans; Isothiocyanates; Kinetics; Male; Morphinans; Protein Binding; Pyrrolidines; Receptors, Opioid, kappa; Tissue Distribution; Up-Regulation

Topics: Animals; Benzomorphans; Binding, Competitive; Brain; Dynorphins; Female; Haloperidol; In Vitro Techniques; Mice; Morphinans; Peptide Fragments; Phencyclidine; Piperidines; Receptors, Opioid

Previous studies of kappa opioid binding sites have suggested heterogeneous binding to this class of opioid receptors. To further investigate kappa receptor heterogeneity, we analyzed the binding properties of various "kappa-selective" ligands in rat brain homogenates. Displacement assays were carried out using [3H]bremazocine in the presence of various displacing ligands under mu and delta receptor-blocked conditions. Homologous displacement of [3H]bremazocine produced "shallow" displacement which best fit a two-site model of drug-receptor interaction. Dynorphin A1-13 and U69,593 exhibited similar biphasic displacement of [3H]bremazocine. Maximal displacement by these ligands, however, represented only approximately 55% of total [3H]bremazocine binding, which suggests the existence of a third component of [3H]bremazocine binding. Biphasic displacement by dynorphin A1-13 was detected in tissue throughout the brain and the spinal cord, whereas the dynorphin-resistant component of [3H]bremazocine binding was uniquely absent in the spinal cord. U50,488H, tifluadom and ethylketocyclazocine appeared to displace from additional, dynorphin-insensitive sites, as their maximal displacement exceeded that seen with either dynorphin A1-13 or U69,593. These results strongly suggest the existence of at least three components of non-mu, non-delta [3H]bremazocine binding in the rat brain: two with differential affinity for dynorphin A1-13 and U69-593 (kappa-1 and kappa-2 sites), and a third (termed here R1) that was further resolved into two binding sites by bremazocine. Preliminary analysis of the R1 component using naloxone revealed one high-affinity site, which may be opiate in nature, and a second site whose binding properties closely resemble those of the sigma receptor described by others.

Topics: Analgesics; Animals; Benzeneacetamides; Benzomorphans; Binding, Competitive; Brain; Culture Techniques; Dynorphins; Ligands; Male; Morphinans; Peptide Fragments; Pyrrolidines; Rats; Rats, Inbred Strains; Receptors, Opioid; Receptors, Opioid, kappa; Tritium

Cell-free translation of mRNA, extracted from NG108-15 cells, was used to examine some properties of the opioid binding sites synthesized in vitro. A monoclonal antiidiotype antibody directed against the delta opioid receptor immunoprecipitated a major band of Mr 51,000. Translational immunoassays of poly[A]+RNA, size fractionated by methylmercury agarose gel electrophoresis, demonstrated that the 51,000 Mr protein specifically immunoprecipitated by the anti-opioid receptor antiidiotype antibodies was coded by a transcript which length was in the 6 to 8 kb range. Displacement binding studies of tritiated ligands (either bremazocine or delta or mu selective peptides) with type selective opioid ligands showed that only one type of opioid binding site was synthesized in vitro. Although the pharmacological profiles of ligands binding to NG108-15 cells were characteristic of the delta receptor type, the de novo synthesized opioid binding site had lost its delta selectivity and showed equal affinity for both the mu and delta but not for the kappa ligands. Similar to our finding using the immunoprecipitation system, size fractionation of the NG108-15 poly[A]+RNA demonstrated that the transcript coding for the "mu-delta" binding site had a length of 6,500 to 7,500 nucleotides.

Topics: Animals; Benzomorphans; Binding, Competitive; Cell Line; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalins; Hybridomas; Mice; Molecular Weight; Morphinans; Oligopeptides; Rats; Receptors, Opioid; Receptors, Opioid, delta; Receptors, Opioid, mu; RNA, Messenger

The purpose of the present investigation was to characterize the mu agonist and kappa antagonist effects of the mixed opioid agonist/antagonist butorphanol. To this end, the effects of butorphanol were examined: 1) alone and in combination with the kappa agonist bremazocine in nontolerant and morphine-tolerant rats responding under a fixed-ratio 30 (FR30) schedule of food presentation, and 2) in rats trained to discriminate 10 mg/kg morphine from saline. Prior to the induction of morphine tolerance, morphine, bremazocine and butorphanol produced dose-dependent decreases in rate of responding under the FR30. In these nontolerant rats, butorphanol failed to antagonize bremazocine's rate-decreasing effects. During the chronic morphine regimen, the dose-effect curve for morphine was shifted to the right of its prechronic position by approximately 0.9 log units, whereas the bremazocine curve was not altered substantially. The butorphanol dose-effect curve, in contrast, was shifted to the right and flattened such that doses which eliminated responding in nontolerant rats, as well as doses approximately 1.0 log unit higher, had no effect on responding. In these morphine-tolerant rats, butorphanol produced a dose-dependent antagonism of bremazocine's rate-decreasing effects. In rats trained to discriminate morphine from saline, butorphanol substituted completely for the morphine stimulus. Unlike morphine, which produced its stimulus effects only at doses that decreased rate of responding, butorphanol substituted for the morphine stimulus at doses that had little or no effect on rate of responding.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Analgesics; Animals; Behavior, Animal; Benzomorphans; Butorphanol; Discrimination, Psychological; Drug Tolerance; Male; Morphinans; Morphine; Narcotic Antagonists; Rats; Receptors, Opioid; Receptors, Opioid, kappa; Receptors, Opioid, mu; Reinforcement Schedule

Delta-opiate receptors have been solubilized with the non-ionic bile salt detergent digitonin from NG108-15 cell membranes and reconstituted into lipid vesicles. Specific opiate binding was restored to soluble receptor preparations after supplementation with a brain lipid extract, and dilution below the effective detergent concentration. Saturable and specific opiate binding was measured for both membrane and vesicle preparations; dissociation constants (Kd) obtained from saturation isotherms of [3H]bremazocine binding were 1.3 and 4.2 nM, respectively. Relative affinity (IC50) values of ligand binding measured for subtype-selective agonists confirmed that a delta-opiate binding site interaction was recovered in vesicle preparations. Changes in agonist binding affinity noted for these experiments were explained by dissociation of the GTP-binding protein Gi from the receptor in detergent. The recovery of solubilized opiate receptors was nearly quantitative, and strictly dependent upon the total brain lipid preparation used in the reconstitution. Ligand binding was incompletely recovered after substituting pure, vesicle-forming phospholipid preparations. [3H]Bremazocine binding was also reconstituted after lectin affinity chromatography of solubilized receptor preparations, using conditions which likely effect the removal of endogenous lipid cofactors. A photoaffinity cross-linking methodology was employed to verify recovery of the delta-opiate receptor after its solubilization from membranes and reconstitution. Two membrane-associated proteins (50 and 70 kDa) were covalently tagged with an azido analog of beta-endorphin(Leu5) in cell membranes and subsequently identified by immunoblotting with antisera directed against this opioid. Labeling of the 50-kDa polypeptide was prevented by coincubating assay samples with a relative excess of (D-Pen2,5)enkephalin. This opioid binding polypeptide was also present in solubilized/reconstituted receptor preparations.

Topics: Animals; Benzomorphans; Binding Sites; Cell Line; Cell Membrane; Chromatography, Affinity; Glioma; Kinetics; Liposomes; Morphinans; Neuroblastoma; Phospholipids; Receptors, Opioid; Receptors, Opioid, delta; Solubility

The behavioral effects of U50,488 [( trans]-3,4-dichloro-N-methyl-N[2-(1- pyrrolidinyl)cyclohexyl]benzeneacetamide), bremazocine, Mr2266 [(-)-5,9-diethyl-2-(3-furylmethyl)-2'-hydroxy-6,7-benzomorphan] and morphine were compared in squirrel monkeys responding under multiple fixed-ratio fixed-interval (FR FI) schedules of food presentation or stimulus-shock termination. Doses of bremazocine (0.001-0.003 mg/kg), U50,488 (0.03-0.1 mg/kg) and Mr2266 (1.0-3.0 mg/kg) that markedly increased overall rates of FI responding maintained by stimulus-shock termination had little effect on or only decreased overall rates of FI responding maintained by food presentation. Each of the kappa opioids decreased FR responding maintained by either consequence. Morphine (0.03-1.7 mg/kg) only decreased responding under all conditions. Pretreatment with Mr2266 (0.1 mg/kg) produced a 10-fold or more rightward shift in the dose-effect functions for morphine under the two multiple schedules and U50,488 under the multiple schedule of food presentation. A 3-fold higher dose of Mr2266 produced an approximately 10-fold rightward shift in the descending portion of the dose-effect functions for U50,488 and bremazocine under the schedule of stimulus-shock termination but did not appreciably alter their rate-increasing effects. Naltrexone (0.1 mg/kg) antagonized the effects of selected doses of morphine or bremazocine on overall rates of responding under the schedule of stimulus-shock termination. In contrast to its effects in combination with morphine, however, naltrexone (0.1-3.0 mg/kg) did not block alterations in patterns of FI responding produced by bremazocine.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Analgesics; Animals; Benzomorphans; Conditioning, Psychological; Dose-Response Relationship, Drug; Electroshock; Male; Morphinans; Morphine; Naltrexone; Pyrrolidines; Receptors, Opioid; Receptors, Opioid, kappa; Saimiri

While it is well established that opiate receptors up-regulate following chronic antagonist treatment in vivo, possible down-regulation following chronic agonist treatment remains controversial. In this study, rats received a continuous seven-day infusion of bremazocine, an opioid drug suggested to be a potent agonist at kappa receptors and an antagonist at mu and delta receptors. Opiate receptor binding was assessed in both cryostat sections and homogenates of rat brain, under conditions selective for mu, delta and kappa sites. Data from both sections and homogenates showed an increase in the capacity of mu binding sites following chronic bremazocine treatment, suggesting that up-regulation of mu receptors had occurred, and that residual ligand from the in vivo treatment had largely been removed. A significant decrease in kappa binding was observed in sections, and experiments using homogenates demonstrated a dramatic loss of high-affinity kappa binding, with an increase in low-affinity binding. There was no apparent alteration in binding to delta receptors. No significant changes were observed following acute injection of bremazocine. Quantitative light-microscopic autoradiography confirmed the results of the binding experiments, and showed that the magnitude of these effects varied between different brain regions. No decrease in kappa binding was seen following chronic administration of the partial kappa agonist nalorphine, indicating that high agonist intrinsic activity is necessary for down-regulation to occur. In addition, chronic co-administration of bremazocine with the partial agonist/antagonist diprenorphine did not cause a significant decrease in kappa binding, implying that diprenorphine can antagonize the down-regulatory effect. These results provide evidence that bremazocine possesses different degrees of intrinsic activity at mu, delta and kappa receptors. They demonstrate that, at least in the case of kappa sites, opiate receptors do show down-regulation following chronic agonist treatment in vivo.

Topics: Animals; Benzomorphans; Binding, Competitive; Endorphins; Male; Morphinans; Rats; Rats, Inbred Strains; Receptors, Opioid; Receptors, Opioid, kappa; Receptors, Opioid, mu

Rat neostriatal slices were superfused with medium containing 0.1 to 30 microM of the dopamine (DA)-releasing agent D-(+)-am-phetamine (AMPH) and the D-2 DA receptor antagonist (-)-sulpiride (10 microM) in the absence or presence of mu-, delta-, and kappa-selective opioids. AMPH dose-dependently enhanced the cyclic AMP production, as measured by its efflux from striatal slices, whereas simultaneous blockade of D-2 DA receptors by (-)-sulpiride strongly potentiated this effect. Both the mu-opioid receptor selective agonist [D-Ala2,MePhe4,Gly-ol5]enkephalin (0.01-3 microM) and the delta-opioid receptor selective agonist [D-Phe2-D-Pen5]enkephalin (DPDPE, 0.01-3 microM) inhibited the cyclic AMP efflux, stimulated by 10 microM AMPH in the presence of (-)-sulpiride, by 70 to 80%. The highly selective kappa-opioid receptor agonist U 50,488 (trans-3,4-dichloro-N-methyl-N-[2-(1-pyrrol-idinyl)- cyclohexyl]benzeneacetamide methanesulfonate hydrate) (0.01-1 microM) had no effect. In contrast, the purported kappa-opioid receptor agonist bremazocine (3-300 nM) inhibited the stimulated adenylate cyclase activity to a similar extent as did [D-Ala2-MePhe4,Gly-ol5]enkephalin and DPDPE. Moreover, the selective irreversible delta-antagonist fentanyl isothiocyanate reversed both the inhibition caused by DPDPE and that caused by bremazocine, whereas the kappa-selective antagonist norbinaltorphimine showed no differences in its potency to antagonize the inhibitory effects of the different opioid agonists. The results indicate that opioids, by activating mu- or delta-, but not kappa-opioid receptors may cause a profound inhibition of adenylate cyclase activity stimulated by activation of (postsynaptic) D-1 DA receptors upon the (presynaptic) release of DA.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenylyl Cyclases; Amphetamine; Animals; Benzomorphans; Corpus Striatum; Cyclic AMP; Dopamine; Drug Interactions; Male; Morphinans; Narcotics; Rats; Rats, Inbred Strains; Receptors, Opioid

Intracerebroventricular or i.p. injections of bremazocine produced a dose-dependent diuretic response and increased glomerular filtration rate in hydrated as well as in nonhydrated rats. The potency and magnitude of the bremazocine-induced diuresis were more pronounced in the nonhydrated group of rats. That bremazocine has a central component of action is deduced from the fact that 0.1 microgram of the opioid administered centrally caused a significant increase in urine output; proportionally, larger doses of bremazocine were required to produce the same diuretic effect when the drug was administered parenterally. Bremazocine did not change the total amount of urinary Na+ and K+ as compared to the saline controls; it increased significantly the free water clearance. The bremazocine-induced diuresis was antagonized in a competitive fashion by 10 mg/kg of naloxone giving further support to the notion that the mechanism of action of bremazocine involves activation of kappa-opioid receptors. Bremazocine injected i.v. to nonanesthetized rats increased mean systemic blood pressure in a dose-dependent manner; the pressor action of the opiate was blocked and prevented by 1 mg/kg of naloxone. In contrast, i.c.v. administration of bremazocine did not change mean systemic blood pressure but produced a dose-related increase in urine output. To determine whether in addition to a central site bremazocine also activates a renal mechanism, experiments were performed in the isolated perfused rat kidney. Bremazocine (0.15-2.5 microM) caused a dose-dependent diuretic response and a significant rise in perfusion pressure as well as in glomerular filtration rate.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Benzomorphans; Blood Pressure; Brain; Diuresis; Female; Glomerular Filtration Rate; Kidney; Kidney Concentrating Ability; Male; Morphinans; Morphine; Naloxone; Rats; Rats, Inbred Strains; Receptors, Opioid; Receptors, Opioid, kappa; Water-Electrolyte Balance

The effect of the kappa-opioid receptor agonist, bremazocine, on plasma oxytocin levels in rats was measured by a sensitive radioimmunoassay. Initially, a decrease in plasma oxytocin levels was seen 30 min after injection. This was in accordance with the bremazocine inhibition of oxytocin release after submaximal electrical stimulation seen in isolated neurointermediate lobes. The initial decrease in plasma oxytocin reversed, and 4 h after injection of bremazocine a 20-fold increase in the oxytocin level was seen. The rise in plasma oxytocin was paralleled by a rise in plasma sodium. The biphasic time course of the plasma oxytocin response can be explained by a combination of an inhibition of oxytocin release from the neurohypophysis and an increased water excretion leading to an elevation in plasma sodium, which may be responsible for the late rise in plasma oxytocin. Down-regulation of the opioid receptors may also contribute to the delayed rise in plasma oxytocin.

Topics: Analgesics; Animals; Benzomorphans; Body Water; Brain Chemistry; Female; In Vitro Techniques; Male; Morphinans; Naloxone; Oxytocin; Pituitary Gland; Rats; Rats, Inbred Strains; Receptors, Opioid; Receptors, Opioid, kappa; Sodium; Time Factors

In the dog cerebral cortex and spinal cord, [3H]bremazocine and [3H]U69593 both bound with high affinity to an apparent single population of binding sites under kappa-selective conditions. In the cortex similar Bmax values for both radioligands in the saturation studies and the high affinity of the kappa-selective agents PD117302 and U69593 for both [3H]bremazocine and [3H]U69593 labelled sites in the competition studies suggested a predominance of U69593-sensitive sites previously described as kappa 1 in the guinea-pig and rat brain. The lower slope values for the inhibition curves of PD117302 and U69593 against [3H]bremazocine but not against [3H]U69593 suggested that [3H]bremazocine could also be binding to a relatively minor proportion of additional, possibly kappa 2, sites while [3H]U69593 would appear to be selective for the kappa 1 site. In contrast, in the dog spinal cord, [3H]U69593 appeared to recognize only a proportion (approximately 35%) of the [3H]bremazocine labelled binding site. The significantly lower affinities and slope values of U69593 and PD117302 against [3H]bremazocine were consistent with the additional sites representing the k2 (benzomorphan) sites previously described in guinea-pig and rat spinal cord. Alternatively, the low (micromolar) affinity of the mu-selective ligand, [D-Ala2, MePhe4, Gly-ol5]enkephalin, implied that these additional sites might not be kappa 2 but possibly a low affinity mu site normally expressed under more physiological conditions.

Topics: Animals; Benzeneacetamides; Benzomorphans; Cerebral Cortex; Dogs; Morphinans; Pyrrolidines; Receptors, Opioid; Receptors, Opioid, kappa; Spinal Cord

In guinea-pig brain, [3H]bremazocine has a binding capacity of 27.2 pmol/g wet tissue, which is statistically different from that of [3H]ethylketazocine (14.7 pmol/g wet tissue) or the sum of the individual binding capacities of mu-, delta-, and kappa-selective ligands (15.0 pmol/g wet tissue). Saturation studies of [3H]bremazocine performed in the presence of unlabelled mu-, delta-, and kappa-blockers still reveal a homogeneous population of binding sites. [3H]Bremazocine under suppressed conditions displays at these sites a Kd of 2.51 nM with a binding capacity of 9.15 pmol/g wet tissue. We have performed the pharmacological characterization of these additional opioid binding sites. Displacement curves measured with a number of opioid substances were all best fitted to a one-site model. The stereoselectivity of these additional sites was demonstrated by using two groups of stereoisomers. Oripavine and benzomorphan opioids were among the most potent drugs at the [3H]bremazocine sites (mu + delta + kappa suppressed). Diprenorphine, bremazocine, cyclazocine, and ethylketazocine displayed apparent affinities constants (1/Ka) of 8.66, 7.57, 21.4, and 38.0 nM, respectively at those sites. The kappa-selective drugs U50488, U69593, PD117302, and tifluadom were inhibitors of the binding of [3H]bremazocine at these sites with apparent affinities of 113, 268, 76.9, and 47.9 nM. All mu- or delta-selective drugs tested in this study have caused weak or no inhibition of the binding. Correlation analyses were done between the different affinities measured at the [3H]bremazocine sites (mu + delta + kappa suppressed) and those observed at the known mu-, delta-, and kappa-sites of the guinea-pig brain.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Benzomorphans; Brain; Cyclazocine; Ethylketocyclazocine; Guinea Pigs; In Vitro Techniques; Kinetics; Male; Membranes; Models, Biological; Morphinans; Muscle, Smooth; Peptides; Receptors, Opioid; Receptors, Opioid, delta; Receptors, Opioid, kappa; Receptors, Opioid, mu; Stereoisomerism

An intraperitoneal injection of the preferential opiate receptor agonist (+/-) bremazocine HCl given to male rough-skinned newts acutely and dose-dependently reduced their spontaneous locomotor activity. Inversely, and contrary to the situation generally observed in other vertebrates, administration of the opiate receptor antagonist naloxone HCl dose-dependently and acutely stimulated locomotion. Given at a behaviorally active dosage, naloxone counteracted the inhibitory effect of bremazocine on locomotion. The behavioral influence of the two substances was observed using two different sampling techniques (continuous recording for 3 minutes: repeated instantaneous sampling for 60 minutes). These data are discussed in view of our current knowledge on the opiate regulation of locomotor activity in vertebrates.

Topics: Animals; Benzomorphans; Dose-Response Relationship, Drug; Injections, Intraperitoneal; Male; Morphinans; Motor Activity; Naloxone; Receptors, Opioid; Receptors, Opioid, kappa; Salamandridae

Pigeons were trained to discriminate a dose of either 0.01 mg/kg of bremazocine or 0.05 mg/kg of fentanyl from water using a two-key drug discrimination procedure. During tests of substitution, the selective kappa-opioid agonists bremazocine, U50, 488 and tifluadom substituted for the bremazocine stimulus, whereas the less selective kappa-opioid agonists ethylketocyclazocine, levallorphan, proxorphan and nalorphine substituted for the fentanyl stimulus. The full mu-opioid agonists fentanyl, morphine, I-methadone and levorphanol, as well as the partial agonists nalbuphine, butorphanol and buprenorphine, substituted for the fentanyl stimulus. Compounds with partial-opioid agonist effects, namely nalbuphine, butorphanol, buprenorphine, proxorphan, levallorphan and nalorphine, produced 50% fentanyl-appropriate responding at doses 25 to 369.2 times smaller than the doses required to decrease response rates to 50% of control values. In contrast, the full mu-opioid agonists fentanyl, morphine, I-methadone and levorphanol produced 50% fentanyl-appropriate responding at doses only 1.3 to 10.9 times smaller than those required to decrease response rates by 50%. During tests of antagonism, both naloxone and Mr2266 produced a dose-dependent attenuation of the stimulus effects of bremazocine and fentanyl, whereas beta-funaltrexamine antagonized the stimulus effects of fentanyl but not bremazocine. Although bremazocine has been reported to have mu-opioid antagonist effects, it failed to antagonize the stimulus effects of the training dose of fentanyl. The present investigation establishes further that pigeons can discriminate selective kappa-opioid agonists from mu-opioid agonists and that in pigeons the classification of numerous opioid compounds on the basis of their kappa-like or mu-like stimulus effects differ from those in rat and monkey. In addition, under the drug discrimination procedure the actions of compounds classified as partial-opioid agonists can be differentiated from those of full mu-opioid agonists on the basis of the ratio of the dose required to engender fentanyl-like stimulus effects to the dose required to reduce response rates.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Animals; Benzodiazepines; Benzomorphans; Columbidae; Cyclazocine; Discrimination Learning; Ethylketocyclazocine; Fentanyl; Morphinans; Naloxone; Narcotics; Pyrrolidines; Receptors, Opioid; Receptors, Opioid, kappa; Receptors, Opioid, mu

Pigeons were trained to discriminate the kappa-opioid agonist bremazocine (BREM) or the mu-opioid agonist fentanyl (FENT) from water. During tests of stimulus substitution, FENT and BREM failed to substitute for each other. The (-)-isomers of cyclazocine, pentazocine and ketocyclazocine substituted for the FENT but not the BREM stimulus. The (+)-isomers of these compounds, as well as the isomers of nallylnormetazocine, failed to substitute for either the FENT or BREM stimulus. In FENT- and BREM-trained pigeons, the (-)-isomers of cyclazocine, pentazocine, nallylnormetazocine and ketocyclazocine were more potent than their respective(+)-isomers in decreasing rates of responding. These results indicate that in the pigeon there is an isomeric separation of the discriminative stimulus properties of cyclazocine, pentazocine and ketocyclazocine and that the FENT-like stimulus effects of these drugs reside in their (-)-isomers. In addition, the present findings establish further that the classification of the discriminative stimulus effects of mu and kappa opioid compounds in the pigeon differ from those in rat and monkey.

Topics: Analgesics; Animals; Benzomorphans; Columbidae; Discrimination Learning; Fentanyl; Morphinans; Racemases and Epimerases; Stereoisomerism; Water

The interaction of the nonselective opioid ligand [3H]bremazocine and of the kappa-opioid [3H]U69593 with the kappa-receptor was investigated in guinea-pig cortical membranes. Each radioligand bound to a single population of high-affinity sites, although [3H]U69593 apparently recognised only 70% of those sites labelled by [3H]bremazocine. Naloxone and the kappa-selective ligands U69593 and PD117302 exhibited full inhibition of the binding of both radioligands. Kinetic analysis demonstrated biphasic rates of association and dissociation for both [3H]bremazocine and [3H]U69593. Detailed analysis of the binding of [3H]U69593 revealed that the fast rate of association was dependent on radioligand concentration, in contrast to the slow rate, which was independent of ligand concentration. Guanylyl-5'-imidodiphosphate (GppNHp) inhibited binding of [3H]U69593; saturation analysis demonstrated that the inhibitory effects of GppNHp resulted in a decrease in affinity without any significant change in binding capacity. GppNHp attenuated the formation of the slow component of [3H]U69593 binding, while accelerating the fast component. The data are consistent with the formation of a high-affinity complex between the kappa-receptor and a guanine nucleotide binding protein. Guanine nucleotides promote the dissociation of this ternary complex and the stabilisation of a lower-affinity state of the receptor.

Topics: Animals; Benzeneacetamides; Benzomorphans; Binding, Competitive; Guinea Pigs; Kinetics; Male; Morphinans; Pyrrolidines; Radioligand Assay; Tritium

The effects of mu-agonists (morphine, fentanyl) and kappa-agonists (U-50,488, U-69,593, bremazocine, nalbuphine, tifluadom) on the electroconvulsive threshold were studied in mice. The threshold could be significantly elevated by all drugs tested in a dose range that was in the order of magnitude of the antinociceptive ED50. Mice tolerant to the antielectroshock effect of morphine still reacted to U-69,593. The antagonism of the anticonvulsant effect by the mu-antagonist naloxone and the kappa-antagonist MR 2266 was receptor-specific only with fentanyl and U-50,488. The other opioid agonists were either antagonized by both drugs (morphine, U-69,593, bremazocine, nalbuphine) or even by the opposite antagonist (tifluadom). A synergistic effect of mu- and kappa-stimulation is assumed for the mediation of the antielectroshock effect of opioid drugs, but drugs with high affinity and intrinsic activity at one receptor type (fentanyl, U-50,488) are obviously able to bring about their antielectroshock effect through the one respective opioid binding site.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Animals; Benzeneacetamides; Benzodiazepines; Benzomorphans; Electroshock; Fentanyl; Male; Mice; Morphinans; Morphine; Nalbuphine; Naloxone; Pyrrolidines; Receptors, Opioid; Receptors, Opioid, kappa; Receptors, Opioid, mu; Seizures

Three novel opioid agonists are described. These compounds were found to bind with high affinity and selectivity to the kappa-opioid receptor. Isolated tissue studies using the field-stimulated mouse vas deferens and guinea-pig ileum preparations confirmed the high agonist potency and naloxone-reversibility of these agents. All three compounds exhibited potent antinociceptive activity in the mouse abdominal constriction model. These compounds should prove useful as tools to investigate kappa-receptor function.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Animals; Benzomorphans; Electric Stimulation; Guinea Pigs; In Vitro Techniques; Mice; Morphinans; Muscle Contraction; Muscle, Smooth; Naloxone; Narcotics; Pyrrolidines; Receptors, Opioid; Receptors, Opioid, kappa

The effects of three kappa opioid agonists namely, bremazocine, tifluadom and U-50,488H were studied on blood pressure and heart rate in urethane-anesthetized normal and bilateral adrenal demedullated rats. Bremazocine (0.2, 0.4 and 0.6 mg/kg i.v.) produced a dose-dependent decrease in heart rate, while only 0.4 mg/kg bremazocine produced marked hypotension. The effect appeared to be long lasting because even at 60 min following drug administration the decreases in both heart rate and blood pressure continued. Bilateral adrenal demedullation did not change bremazocine-induced fall in blood pressure but the bradycardia was partially blocked. Tifluadom (0.1-0.4 mg/kg i.v.) produced an initial arrest of heart beat followed by bradycardia which recovered in about 60 min. Except for a very transient fall soon after drug administration no significant effect was observed on blood pressure. In adrenal demedullated rats, tifluadom induced initial arrest of heart was not affected but the subsequent bradycardia was blocked. U-50,488H (0.2, 0.4 and 0.6 mg/kg i.v.) produced dose-dependent bradycardia and hypotension both of which were blocked following bilateral adrenal demedullation. Naltrexone methylbromide (MRZ 2663 BR), a quaternary opioid antagonist, injected 5 min prior to U-50,488H, blocked its cardiovascular effects. The results suggest that kappa opioid agonists given i.v. depress cardiovascular system and these effects are mediated through the adrenal medulla and peripheral opioid receptors. The differential effects of kappa opioid agonists on blood pressure and heart rate suggest that either the three kappa agents interact differentially at the kappa opioid receptors or the subtypes of receptors for the kappa opioid exist.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Adrenal Medulla; Animals; Benzomorphans; Blood Pressure; Cardiovascular System; Dose-Response Relationship, Drug; Heart Rate; Male; Morphinans; Naltrexone; Narcotic Antagonists; Pyrrolidines; Rats; Rats, Inbred Strains; Tape Recording

Intravenously administered bombesin lowered basal PRL levels in conscious male rats and prevented the morphine, bremazocine and stress-induced PRL secretion. The same dose of bombesin had no effect on PRL levels in alpha-methyl-p-tyrosine pretreated rats and did not affect haloperidol-stimulated PRL release. These results show that bombesin given intravenously acts as an inhibitor of PRL secretion and suggests that it does not act on the lactotrope itself but rather by an increase of the inhibitory dopaminergic tone.

Topics: alpha-Methyltyrosine; Analgesics; Animals; Benzomorphans; Bombesin; Haloperidol; Male; Methyltyrosines; Morphinans; Prolactin; Rats; Rats, Inbred Strains; Reference Values; Restraint, Physical; Stress, Psychological; Tyrosine 3-Monooxygenase

In three experiments we examined the analgesic potency of kappa opioid receptor agonists in 2- and 16-day-old rats. Ethylketocyclazocine (1-50 mg/kg) produced similar dose- and time-dependent increases in the latency to retract a hind paw from a noxious thermal stimulus in rats of both ages. Bremazocine (0.001-10 mg/kg), a kappa agonist with reported antagonist activity at mu receptors, was also effective in producing analgesia in 2-day-old rats. The dose-effect relationship for bremazocine was nonmonotonic. Bremazocine analgesia (0.1 mg/kg) was reversed by both naltrexone and MR2266, a putative kappa opioid antagonist. These results are discussed in terms of the functional integrity of a kappa analgesic system in the developing rat.

Topics: Analgesia; Analgesics; Animals; Animals, Newborn; Benzomorphans; Cyclazocine; Ethylketocyclazocine; Female; Male; Morphinans; Naltrexone; Narcotic Antagonists; Rats

It has been found that bremazocine predominantly enhances the flexor reflex of the right hind limb, without affecting the left limb. The similarity of the visual data measuring the asymmetry of the pose and the asymmetry of EMG responses to painful stimulation has a high probability. No correlation between the asymmetry of the pose and EMG extensor activity has been found.

Topics: Analgesics; Animals; Benzomorphans; Electromyography; Hindlimb; Male; Morphinans; Muscles; Rats; Reflex; Spinal Cord

Four experiments were performed to evaluate a possible opioid involvement in the regulation of sexual behavior (amplectic clasping of a female) in intact adult male rough-skinned newts (Taricha granulosa) during the breeding season. It was found that an ip injection of bremazocine, a kappa-receptor opiate agonist, can markedly reduce sexual activity and that an ip injection of naloxone can reverse this inhibition in a dose-dependent fashion. In contrast, in male newts that were sexually inactive before treatment, injections of naloxone failed to induce sexual behavior, suggesting that opioid mechanisms do not normally exert a tonic inhibition of amphibian sexual behavior. In addition, an injection of ethylketocyclazocine (another kappa-receptor agonist), but not morphine (a mu-receptor agonist) suppressed sexual behaviors of male newts. These results indicate that opioid mechanisms that include kappa-type opioid receptors may contribute to the regulation of sexual behavior in nonmammalian vertebrates.

Topics: Analgesics; Animals; Benzomorphans; Cyclazocine; Ethylketocyclazocine; Male; Morphinans; Morphine; Naloxone; Receptors, Opioid; Salamandridae; Sexual Behavior, Animal

1. The autoradiographic distribution of kappa opioid receptor binding sites in human brain was examined using two radiolabeled probes, namely [3H]U69,593 and [3H]bremazocine. 2. [3H]U69,593 binding was performed in the absence of blockers for other sites, while [3H]bremazocine binding was investigated in the presence of saturating concentrations of mu and delta blockers to ensure selective labeling of kappa opioid receptors. 3. Our results show that the autoradiographic distribution of [3H]U69,593 and [3H]bremazocine (plus blockers) binding sites is identical, with high densities of sites found in deep cortical layers and claustrum. 4. This indicates that [3H]U69,593 is a highly selective ligand of the kappa opioid receptor type.

Topics: Autoradiography; Benzeneacetamides; Benzomorphans; Binding Sites; Brain; Humans; In Vitro Techniques; Morphinans; Pyrrolidines; Receptors, Opioid; Receptors, Opioid, kappa; Tissue Distribution

D-Ala2, D-Leu5-enkephalin (DADLE) and dynorphin1-13 (Dyn1-13) inhibited striatal adenylate cyclase activity, both basal and dopamine-stimulated (DA), in rats and guinea pigs. The kappa-agonists bremazocine (BRZ), U-50,488 (trans-3,4-dicloro-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl]- benzeneacetamide), and U-69,593 (5 alpha, 7 alpha 8 beta)-(-)-N-methyl-N-(7-(1-pyrrolidinyl-1-oxaspiro (4.5)dec-8yl) benzeneacetamide inhibited only the basal adenylate cyclase activity, and such an effect was restricted to guinea pig striatum, an area known to contain a high density of kappa-binding sites. Moreover, BRZ was found to antagonize the inhibitory effect of both DADLE and Dyn1-13 in rat striatum.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Adenylyl Cyclases; Analgesics; Animals; Benzeneacetamides; Benzomorphans; Corpus Striatum; Dynorphins; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalins; Guinea Pigs; Male; Morphinans; Peptide Fragments; Pyrrolidines; Rats; Rats, Inbred Strains; Receptors, Opioid

The effects of mu, delta and kappa opioid receptor agonists were examined on evoked field potentials in brain slices prepared from rat hippocampus. The effects of the mu-selective opioid peptide [D-Ala2, NMe-Phe4, Met(O)5ol]enkephalin (FK 33-824) and the delta-selective peptide [D-Pen2, D-Pen5]enkephalin (DPDPE) were qualitatively and quantitatively similar. Both increased the amplitude of evoked population spike responses when perfused in low nanomolar concentrations in a fashion consistent with what has been previously reported for other opiate agonists such as morphine. The kappa-selective agonists bremazocine and U-50, 488H were without effect upon evoked responses at concentrations as high as 10 microM. Bremazocine, but not U-50, 488H, proved to be an extremely potent antagonist of responses to both mu- and delta- selective agonists. Moreover, bremazocine was considerably more potent in antagonizing responses to FK 33-824 than DPDPE, which supports the hypothesis that FK 33-824 and DPDPE act via different receptors. Thus, although bremazocine is an agonist at kappa receptors, it appears to act as an antagonist at other opioid receptor sites.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Animals; Benzomorphans; D-Ala(2),MePhe(4),Met(0)-ol-enkephalin; Electric Stimulation; Electrodes; Enkephalin, D-Penicillamine (2,5)-; Enkephalins; Hippocampus; In Vitro Techniques; Male; Morphinans; Pyrrolidines; Rats; Rats, Inbred Strains; Receptors, Opioid; Receptors, Opioid, delta; Receptors, Opioid, mu

Injections of an opioid agonist (bremazocine) and/or an antagonist (quadazocine) were given to heifers during the luteal or follicular phase of the oestrous cycle. Quadazocine was injected (210 mg/injection) three times at 2-h intervals, and bremazocine was injected (0.45 mg/injection) every 15 min for 6 h. Blood samples were taken every 15 min beginning 6 h before treatments started and continued for 18 h. LH secretion patterns were not affected by quadazocine in the luteal-phase heifers, but quadazocine and bremazocine had marked effects during the follicular phase. Quadazocine increased LH secretion by increasing peak height but not peak frequency. Bremazocine decreased LH secretion through both peak height and frequency. This decrease was of greater magnitude than the increase due to quadazocine. When quadazocine and bremazocine were given together, these effects were cancelled and none of the effects carried over into the bleeding period after treatments stopped. No apparent interruption of follicular maturation was detected since all follicular-phase heifers were detected in oestrus at normal intervals. We conclude that heifers in this experiment did not have an opioid-mediated mechanism for progesterone suppression of LH but that an opioid mechanism for modulating LH does exist during the follicular phase.

Topics: Animals; Azocines; Benzomorphans; Cattle; Estrus; Female; Follicular Phase; Luteinizing Hormone; Morphinans; Narcotic Antagonists; Secretory Rate

The increased urine output after injection of the kappa agonist bremazocine was determined in rats of various body weights. Rats of all sizes were sensitive to the diuretic effect of bremazocine. The total diuretic effect was greater in heavier animals. The urine output plateaued between 4-5 ml/100 g body weight irrespective of body weight at the higher doses of bremazocine.

Topics: Analgesics; Animals; Benzomorphans; Body Weight; Dose-Response Relationship, Drug; Male; Morphinans; Rats; Urination

Active opioid binding sites, that retain the ability to bind tritiated agonists have been obtained in good yield in digitonin/NaCl/Mg2+ extracts of morphine protected bovine striatal membranes. Ligand protection of binding sites and the presence of Mg ions were found to be absolute requirements for agonist binding in this solubilized opioid receptor preparation. Soluble preparations contained a ratio of mu: delta:kappa similar to that in the membranes.

Topics: Analgesics; Animals; Benzomorphans; Cattle; Cell Membrane; Corpus Striatum; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine; Enkephalins; Kinetics; Morphinans; Oligopeptides; Receptors, Opioid; Receptors, Opioid, delta; Receptors, Opioid, kappa; Receptors, Opioid, mu

The effects of bremazocine and U-50,488, two selective opioid kappa receptor agonists, and the preferential mu receptor agonist morphine on the secretion of PRL and GH were compared in conscious male rats bearing permanent right atrial cannulae for serial blood sampling and drug delivery. All three opioids stimulated PRL secretion in a dose-related manner, but the kappa agonists differed from morphine in several respects. They were considerably more potent than morphine in triggering a PRL response, but were unable to elevate PRL levels to more than 100 ng/ml, whereas morphine, at the highest dose (4.5 mg/kg), induced an almost twice larger response. Also their PRL-releasing effect was inhibited more strongly by the preferential kappa receptor antagonist Mr-2266 than by naloxone, whereas Mr-2266 and naloxone, which are equipotent as antagonists of the mu receptors, were equipotent in suppressing the PRL-stimulating effect of morphine, a mu agonist. In a complete contrast to morphine, which effectively stimulated GH secretion, the kappa agonists had no effect on GH release at lower doses and suppressed it at higher doses. It is concluded that the PRL-releasing effect of the kappa agonists is mediated by the kappa receptors which may participate with the mu receptors in regulation of PRL secretion by opioids. The GH-inhibiting effect of the kappa agonists requires further clarification.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Animals; Benzomorphans; Dose-Response Relationship, Drug; Growth Hormone; Injections, Intravenous; Male; Morphinans; Morphine; Naloxone; Prolactin; Pyrrolidines; Rats; Rats, Inbred Strains; Receptors, Opioid; Receptors, Opioid, kappa; Time Factors

An analysis of the GH release-inhibiting action of the opioid kappa receptor agonists bremazocine and U-50,488, established earlier, was attempted by testing the agonists against activation of GH secretion by morphine or clonidine in male rats bearing right atrial cannulae for serial blood sampling and drug delivery. Both kappa agonists inhibited the effect of subsequent administration of clonidine in a dose-related manner. Bremazocine was approximately ten times more potent than U-50,488, a ratio corresponding to the known affinities of the two compounds for the kappa receptors. The inhibiting action of bremazocine was more strongly reversed by the preferential kappa receptor antagonist Mr-2266 than by naloxone, neither of which interfered with the GH-stimulating effect of clonidine. Bremazocine, however, did not alter the activation of GH secretion by exogenous growth hormone releasing factor. Thus, the inhibiting effect of bremazocine and probably U-50,488 seems to be derived from stimulation of the kappa receptors which in turn activates a GH release inhibiting mechanism of unknown identify which, however, does not involve release of somatostatin. Both kappa agonists also inhibited the effect of morphine, but in this case U-50,488 was approximately hundred times less effective than bremazocine. Since bremazocine and U-50,488 are antagonists of the delta receptors, which seem to mediate the GH-releasing effect to morphine, their inhibiting effect in this instance may be related to this property rather than to an action on the kappa receptors. Bremazocine, but not U-50,488, was also highly effective in inhibiting stimulation of PRL secretion by morphine.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Animals; Benzomorphans; Clonidine; Growth Hormone; Injections, Intravenous; Male; Morphinans; Morphine; Prolactin; Pyrrolidines; Rats; Rats, Inbred Strains; Receptors, Opioid; Receptors, Opioid, kappa; Serotonin; Time Factors

Bremazocine dose-dependently induced backwards walking behavior in rats after its SC injection. Only the (-) but not the (+) enantiomer induced backwards walking. Pretreatment with either naloxone or MR 2266 reduced the bremazocine-induced backwards walking. MR 2266 was at least ten times more potent than naloxone. These findings suggest that bremazocine-induced backwards walking is mediated via an agonistic action of the drug with opioid kappa receptors. The data may contribute to the discussion concerning opioid kappa receptors and the psychotomimetic effects of some opioid analgesic drugs.

Topics: Animals; Benzomorphans; Brain; Hallucinogens; Locomotion; Male; Morphinans; Naloxone; Rats; Rats, Inbred Strains; Receptors, Opioid; Receptors, Opioid, kappa

[3H]Bremazocine, in the presence of saturating concentrations of mu and delta receptor blocking agents, was used to label putative kappa opiate binding sites in rat brain. The binding of [3H]bremazocine under these conditions was completely displaced with high affinity by U-50488H and dynorphin1-17, and the potency of a series of opiate ligands was consistent with an action at kappa receptors. Therefore, [3H]bremazocine, in the presence of mu and delta blockers, was used to localize U-50488H-displaceable kappa binding sites by autoradiography. A distribution different from that of mu and delta receptors was seen, with levels highest in the claustrum, striatum, medial preoptic area, suprachiasmatic nucleus, medial amygdala and superior layer of the superior colliculus. The results show that the U-50488H-displaceable kappa sites have a distinct distribution which is discussed in terms of the possible functional roles of kappa receptors.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Animals; Autoradiography; Benzomorphans; Binding Sites; Binding, Competitive; Brain; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalins; Male; Morphinans; Pyrrolidines; Rats; Rats, Inbred Strains; Receptors, Opioid; Receptors, Opioid, kappa

The agonist and antagonist activity of bremazocine at opioid receptors in the guinea-pig myenteric plexus preparation was determined in untreated tissues and in tissues in which either mu-9 or kappa-opioid receptors were blocked preferentially. After pretreatment of the tissue with beta-funaltrexamine for 90 min followed by washing out, the IC50 value of the selective mu-ligand [D-Ala2,MePhe4,Gly-ol5]enkephalin was increased 67 fold whereas the IC50 values of the selective kappa-ligand U-69,593 and of the non-selective kappa-ligand bremazocine were not significantly changed. In this experimental design bremazocine acted only on kappa-receptors. After pretreatment of the tissue with beta-chlornaltrexamine and 10 microM of the mu-ligand for 30 min followed by washout, the IC50 value of the mu-ligand was increased 2 fold whereas the IC50 value of the selective kappa-ligand was increased 32 fold and that of bremazocine 62 fold. Under these experimental conditions, it was shown that bremazocine is an antagonist against [D-Ala2,MePhe4,Gly-ol5]enkephalin at the mu-receptor (Ke = 1.6 nM). The residual agonist activity of bremazocine is at the kappa-receptor. In naive myenteric plexus preparations the mu-antagonist activity of bremazocine cannot be demonstrated because its potency at the kappa-receptor is very high. This dual action may be of importance for the responses of bremazocine in other peripheral and central tissues.

Topics: Animals; Benzomorphans; Electric Stimulation; Guinea Pigs; In Vitro Techniques; Morphinans; Myenteric Plexus; Naltrexone; Narcotic Antagonists; Receptors, Opioid; Receptors, Opioid, kappa; Receptors, Opioid, mu

The effects of bremazocine on memory processes were studied in DBA/2 mice tested in a passive avoidance apparatus. In a first set of experiments, memory impairments following immediately posttraining bremazocine administration (0.025 and 0.05 but not 0.01 mg/kg), and memory improvements following immediately posttraining administration of the kappa-opioid-receptor antagonist MR-1452 (1.0 and 2.0 but not 0.5 mg/kg), were observed. No effect was evident when the drugs were injected starting 120 min after training. In a second set of experiments, the effects of bremazocine (0.05 mg/kg) were antagonized by a per se ineffective dose of MR-1452 (0.5 mg/kg), suggesting involvement of kappa-opioid-receptors. In a third set of experiments, the effects of bremazocine were enhanced by a per se ineffective (15 min) immobilization stress, and were decreased by familiarization with the passive avoidance apparatus. The results are discussed in terms of attenuation of emotionality following bremazocine administration.

Topics: Analgesics; Animals; Avoidance Learning; Benzomorphans; Male; Mice; Mice, Inbred DBA; Morphinans; Narcotic Antagonists; Restraint, Physical; Stress, Psychological

Receptor binding experiments with masking of mu- and delta-receptors in the presence of an excess of unlabelled dihydromorphine and [D-Ala2, D-Leu5]enkephalin have been carried out. They indicate that 12 to 17% of original high affinity binding of [3H]Mr 2034 [-)(1R,5R,9R,2"S)-5,9-dimethyl-2-tetrahydrofurfuryl-2'-hydroxy-6, 7-benzomorphan hydrochloride), an opioid kappa-agonist, could then be detected as kappa-receptor sites in brain membranes both from untreated rats and from rats pretreated with naloxazone, too. Because of the irreversible blockade of the high affinity binding sites by naloxazone (naloxone hydrazone) treatment, the masking effects of mu- and delta-selective ligands seem to be mediated by the low affinity binding sites of these opioid receptor types. As could be shown before with [3H]Mr 2034, another kappa-agonist, [3H]bremazocine does not seem to be affected in its binding properties by naloxazone treatment of the rat in vivo. Displacement studies with several opioid agonists and antagonists, [3H]Mr 2034 and [3H]ethylketocyclazocine as radioligands in brain membranes from naloxazone treated rats and untreated controls provided further support for the evidence of two different kappa-receptors.

Topics: Animals; Benzomorphans; Binding, Competitive; Cyclazocine; Dihydromorphine; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Ethylketocyclazocine; In Vitro Techniques; Kinetics; Male; Morphinans; Naloxone; Rats; Receptors, Opioid; Receptors, Opioid, kappa

In vivo binding affinities of three benzomorphans, Win 44,441-3, bremazocine and MR 2266, were determined at the mu, delta and kappa types of opioid binding sites in rat brain, using an ex vivo labeling technique. The receptor occupancy of the benzomorphans and of previously tested diprenorphine were compared with their activities in increasing urine output (agonist ED50: bremazocine) or inhibiting bremazocine-induced diuresis (antagonist ID50: Win 44,441-3, MR 2266 and diprenorphine). The agonist, bremazocine, bound (in order of decreasing affinity) to the kappa approximately equal to mu greater than delta binding sites, and it's pharmacological effects appeared in the dosage range of kappa and mu binding. In order to positively identify which receptor type is responsible, the potency of the three antagonists to block the effects of bremazocine were compared to their ability to occupy the individual sites in vivo. A fractional occupancy of 0.5 would be expected at the ID50 if one assumes a linear relationship between receptor occupancy by the antagonist and the antagonistic effect. Such a linear relationship was observed for the three antagonists only at the kappa site, whereas variable occupancies were observed at the mu and delta sites. These results support the previously proposed hypothesis that kappa receptors mediate the effects of benzomorphan opioid drugs on urine flow.

Topics: Animals; Benzomorphans; Brain; Diprenorphine; Diuresis; Morphinans; Rats; Receptors, Opioid; Receptors, Opioid, delta; Receptors, Opioid, kappa; Receptors, Opioid, mu

Long-term treatment for more than 3 months with a central nervous system (CNS)-active drug, the opioid agonist bremazocine, at a dose of 1 mg/kg per day elicited an 80% inhibition of the volume of the subcutaneously transplanted rat prostate adenocarcinoma Dunning R3327H. Whereas, under this therapy, prostate tumour and prostatic weights were decreased, testes and pituitary weights remained normal. Bremazocine inhibited not only the growth of freshly transplanted tumours but also that of well-grown Dunning prostate carcinomas since, after 41 days of treatment, such tumours showed a volume inhibition of 52%. In these experiments bremazocine decreased LH and testosterone plasma levels significantly. Bremazocine, therefore, probably acts mainly through suprapituitary CNS-opiate receptor sites, which indirectly, rather than locally, mediate LH inhibition. Indeed, no specific receptors for bremazocine could be found in the Dunning tumour, which makes a local action of bremazocine in this tissue unlikely. the efficient tumour growth inhibition through the supra-pituitary action of bremazocine makes such opiate drugs, which lack respiratory and side effects due to improper use, of potential interest for treatment of prostatic tumours.

Topics: Adenocarcinoma; Animals; Benzomorphans; Central Nervous System; Luteinizing Hormone; Male; Morphinans; Organ Size; Prostate; Prostatic Neoplasms; Rats; Rats, Inbred Strains; Testosterone

The effects of the benzomorphan analogue bremazocine on locomotor activity were studied in DBA/2 (DBA) and C57BL/6 (C57) mice. Three sets of experiments were carried out. In a first set, bremazocine depressed activity in both strains, but DBA mice were more sensitive to the effects of the drug. In a second set of experiments the effects of bremazocine were antagonized by a per se ineffective dose (0.5 mg/kg) of the selective kappa-opioid receptor antagonist MR 1452, showing that they were mediated through an interaction with this sub-population of opioid receptors. In a third set of experiments the effects of bremazocine were enhanced by the administration of a per se ineffective dose (0.025 mg/kg) of haloperidol, showing the involvement of dopaminergic mechanisms.

Topics: Analgesics; Animals; Benzomorphans; Haloperidol; Male; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; Morphinans; Motor Activity; Narcotic Antagonists; Receptors, Dopamine; Species Specificity

In prehydrated rats, the administration of kappa-opiate agonists such as bremazocine, ethylketocyclazocine or compound Upjohn-50,488 produced a dose-dependent increase in urine output and decreased the concentration of Na+ and K+ in the urine as compared to that of saline-treated rats. The diuretic effect of bremazocine lasted at least 3 h. The increase in urine output was independent of the hydration state of the rat since in non water-loaded animals, bremazocine produced proportionally as much diuresis and a decrease in the output of urine electrolytes of about the same magnitude as that observed in the prehydrated animals treated with the opioid. In contrast to the diuretic action of kappa-opiate agonists, the administration of antagonists with high affinity for the kappa-opiate receptor (Win 44,441 or Mr 2266) decreased dose dependently the output of urine and reduced very significantly the total output of Na+ and K+. Whereas 2 mg/kg naloxone did not block the bremazocine-induced urinary effects, 1 mg/kg Win 44,441 or Mr 2266 antagonized competitively the renal activity of bremazocine. The results are interpreted to suggest that the kappa-opiate receptor may be involved in the regulation of fluid and electrolyte balance.

Topics: Animals; Azocines; Benzomorphans; Cyclazocine; Diuresis; Ethylketocyclazocine; Female; Injections, Intraperitoneal; Morphinans; Morphine; Naloxone; Potassium; Rats; Rats, Inbred Strains; Receptors, Opioid; Sodium; Water-Electrolyte Balance

Autoradiograms of rat brain sections were compared obtained from animals receiving a tritiated drug through intravenous injection or from precut sections incubated in vitro. The benzomorphan analogue 3H-(-)-bremazocine was used as ligand and its distribution to all different opioid binding sites was followed. Although the general distribution of opioid binding sites visualized on 3H-LKB ultrofilms was independent of the methodological approach used, the maximal number of such sites (Bmax) was greater in brain sections incubated in vitro than after in vivo drug application. Since the number of binding sites is highly dependent on the particular incubation condition used, this finding has no further relevance.

Topics: Animals; Autoradiography; Benzomorphans; Binding Sites; Brain; In Vitro Techniques; Morphinans; Rats; Receptors, Opioid

Bremazocine at doses of 0.01 mg kg-1 s.c., and higher, increased serum corticosterone concentration several-fold in rats. The increase occurred within 20 min, was maximum at 40-60 min and subsided after 120 min. Pretreatment with naloxone (1-10 mg kg-1 s.c.) antagonized the corticosterone increase. These data support the view that kappa-opioid agonist activity of bremazocine mediated the corticosterone increase.

Topics: Animals; Benzomorphans; Corticosterone; Dose-Response Relationship, Drug; Male; Morphinans; Naloxone; Rats; Rats, Inbred Strains; Receptors, Opioid; Receptors, Opioid, kappa

Some in vivo agonist and antagonist properties of the putative k-compound bremazocine were characterized in rats. Bremazocine, at doses from 0.015-32 mg/kg i.p., delayed nociceptive reaction on a 55 degrees C hot-plate with a dose-response curve not readily fitting a single straight line; this effect was antagonized by high doses of naloxone. In the same rats bremazocine did not delay the intestinal transit of a charcoal meal fed 5 min earlier and prevented morphine-induced constipation. This antagonism appeared to be opioid-specific and competitive, with apparent pA2 value 8.56. Catatonia induced by etorphine (0.004 mg/kg s.c.) and constipation induced by etorphine (0.004 mg/kg s.c.) and D-Ala2-D-Leu5-enkephalin (0.1 mg/kg i.p.) were completely antagonized by bremazocine (0.03-8 mg/kg i.p.). Antinociception induced by morphine (10 mg/kg i.v.) and etorphine (0.004 mg/kg s.c.) was only partly prevented. Naloxone (1 mg/kg) and bremazocine (0.015-1 mg/kg i.p.) precipitated a withdrawal syndrome, evaluated as jumping frequency, in rats rendered dependent to morphine. These data suggest the involvement of more than one opioid receptor population in bremazocine action in vivo.

Topics: Analgesics; Animals; Benzomorphans; Catatonia; Constipation; Dose-Response Relationship, Drug; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Etorphine; Gastrointestinal Motility; Humans; Male; Morphinans; Morphine; Naloxone; Rats; Substance Withdrawal Syndrome; Substance-Related Disorders

The hypothesis that the nociceptive state and opiate-induced antinociception are generally regulated by Ca2+ brain levels has been tested. In this context, the effects of intracerebroventricular injections of CaCl2 (0.1-0.5 mumol), D600 (5.0-10.0 micrograms) and EGTA (0.5-1.0 mumol) on ethylketocyclazocine (EKC), ketocyclazocine (KC), Mr-2033, pentazocine (PTC), bremazocine (BMC) and SKF 10,047-induced antinociception were investigated in the mouse tail immersion test. Simultaneous treatment with either D600 or EGTA resulted in a significant and dose-related enhancement in the activities of the kappa-agonists: EKC, KC and Mr-2033, whilst the activities of PTC, BMC and SKF 10,047 remained unchanged. CaCl2 readily blocked the activities of all benzomorphans tested except that of SKF 10,047 against which CaCl2 was less effective. In addition a dose-related hyperalgesia was observed when CaCl2 was given alone. Although the results obtained from the kappa-agonists and CaCl2 per se support the hypothesis in question, data obtained from PTC, BMC and SKF 10,047 tends to oppose it. Additionally the present results taken together indirectly substantiate the notion that benzomorphan-induced analgesia may involve different opiate-sensitive neuronal substrates.

Topics: Analgesia; Analgesics; Analgesics, Opioid; Animals; Benzomorphans; Calcium; Calcium Channel Blockers; Cyclazocine; Egtazic Acid; Ethylketocyclazocine; Gallopamil; Male; Mice; Morphinans; Pentazocine; Phenazocine

The effects of various opioid agonists and antagonists on urination were studied in the normally hydrated rat. Two kappa agonists, U-50,488H and proxorphan, markedly increased urination. The increased urination produced by U-50,488H was antagonized by opioid antagonists in a potency order which indicated that the effects were due to an action at kappa opioid receptors. Mu agonists decreased urination and were blocked by low doses (0.01 and 0.1 mg/kg) of naloxone, whereas kappa agonists increased urination and were only blocked by a high dose (10 mg/kg) of naloxone. The diuretic effects of U-50,488H and ketazocine, but not proxorphan and bremazocine, were reduced by morphine, consistent with the idea that proxorphan and bremazocine have morphine antagonist activity. Water deprivation produced a shift to the right for the dose-effect curve for bremazocine-induced diuresis. Kappa agonists were ineffective in increasing urination in Brattleboro rats that were homozygous for diabetes insipidus, whereas mu agonists were still effective in decreasing urination. The data are consistent with the hypothesis that kappa agonists inhibit release of vasopressin from the neurohypophysis and this decrease in vasopressin release leads to increased urination. The effects of opioids on urination in the normally hydrated rat can be extremely useful in classifying the activities of opioid on mu and kappa receptors in vivo.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Animals; Benzomorphans; Cyclazocine; Diuresis; Ethylketocyclazocine; Male; Morphinans; Naloxone; Narcotic Antagonists; Narcotics; Piperidines; Pyrrolidines; Rats; Rats, Inbred Strains; Receptors, Opioid; Receptors, Opioid, kappa; Receptors, Opioid, mu; Urination

The benzomorphan analogue bremazocine has been shown to be a potent analgesic with a low dependence liability and with no respiratory depressant effects in preliminary pharmacologic screening. As this compound may prove to be of potential interest to anesthesiologists, it was tested in the conscious dog in increasing doses (1, 5, 10, 20, and 40 micrograms/kg) while blood pressure, heart rate, respiratory rate, arterial blood-gas tensions, electrical cortical activity, and somatosensory-evoked potentials (SS-EVP) were recorded. Bremazocine had no significant cardiovascular or respiratory effects. Power spectral analysis of the EEG showed a dose-related increase of power in the theta to delta band. SS-EVP was characterized by a decrease in amplitude of the P45 and P70 waves and an increase in the latency of the P 100 peak. EEG and peak latency changes could be reversed by the kappa-specific antagonist Mr 2266 (20 micrograms/kg), but not by the classic opiate antagonist naloxone (20 micrograms/kg). It is suggested that kappa receptor sites distinct from those interacting with common opioids (mu) are responsible for the observed changes associated with bremazocine.

Topics: Analgesics; Animals; Benzomorphans; Blood Gas Analysis; Blood Pressure; Body Temperature; Dogs; Drug Administration Schedule; Evoked Potentials, Somatosensory; Heart Rate; Morphinans; Receptors, Opioid; Receptors, Opioid, kappa; Respiration

The effects of bremazocine on drinking and eating behaviour were studied in non-deprived mice and 12 hr food and water deprived rats. In both species, bremazocine (1-8 mg/kg) produced substantial increase in fluid intake (dose-dependent in rats). The concomitant alterations of food intake were less pronounced and opposite: increase in mice, decrease in rats. Morphine (1.25-10.0 mg/kg) invariably inhibited eating and drinking behaviour in mice. The afore-said effects of the kappa-receptor agonist bremazocine are not affected by naloxone which attenuates the effects of the mu-receptor agonist morphine.

Topics: Analgesics; Animals; Benzomorphans; Dose-Response Relationship, Drug; Drinking; Eating; Female; Mice; Mice, Inbred Strains; Morphinans; Rats; Rats, Inbred Strains; Receptors, Opioid; Species Specificity

The respiratory depression and analgesia induced by a kappa-agonist, bremazocine, were studied by the ED50 ratios and the apparent pA2 values of the interaction with naloxone. The ratio between the ED50 in respiratory depression and analgesia (i.c.v.) was very high compared to that of other mu- and delta-agonists, indicating its small activity on the respiratory center. pA2 values were 7.33 and 7.20 for analgesia and respiratory depression, respectively, after i.c.v. administration. After s.c. injection, the pA2 value for analgesia was 6.16. The results confirm the involvement of different opiate receptors in the mediation of supraspinal and spinal analgesia. The results suggest that bremazocine may induce analgesia by interacting with different types of receptors located at different levels of the CNS. This property and the low activity on the respiratory center make drugs like bremazocine good candidates in the search for safer opiate analgesics.

Topics: Analgesia; Analgesics; Animals; Benzomorphans; Drug Antagonism; Morphinans; Naloxone; Rats; Receptors, Opioid; Receptors, Opioid, kappa; Respiration

Morphine and other opioid compounds such as the new benzomorphan derivative, bremazocine, inhibit the secretion of luteinizing hormone in rats of both sexes (1, 2, 3, 4). The aim of our work was to compare in rats the LH-secretion inhibiting properties of bremazocine, a putative opiate kappa agonist (5), with those of the mu agonist morphine. Acute administration of bremazocine (0.005 - 1 mg/kg s.c.) or of morphine (10 - 20 mg/kg s.c.) diminished serum LH levels and spontaneous ovulation in female rats in a dose-dependent manner. Chronic treatment with bremazocine significantly diminished LH and testosterone secretions in male rats which in turn led to a fall in weight of the prostate gland; prolactin and FSH secretions were not influenced significantly. The mu-antagonist naloxone, which increases LH release in rats, in acute experiments significantly antagonized the inhibiting effect of morphine, but not that of bremazocine on LH secretion. Neither the basal nor the LHRH-stimulated secretion of LH in pituitary cell cultures were changed by bremazocine (10(-11) to 10(-5) M), however the release of LHRH-like activity from hypothalamic fragments was significantly impaired by 10(-7) M bremazocine. In conclusion, the data presented here show that bremazocine is a new non-morphine-like opioid agonist which selectively inhibits LH release in rats.

Topics: Animals; Benzomorphans; Castration; Cells, Cultured; Female; In Vitro Techniques; Luteinizing Hormone; Male; Morphinans; Morphine; Naloxone; Ovulation; Pituitary Gland; Rats; Rats, Inbred Strains; Reproduction; Testis; Time Factors

Cultured hippocampal pyramidal cells responded to field stimulation with a short latency excitation followed by a long-lasting inhibition. This sequence was transformed into a bursting response by bath application of 10(-8) M FK 33-824, 10(-6) M (D-Ala)2(D-Leu)5-enkephalin and 10(-5) M bremazocine. Bremazocine and ethylketocyclazocine stereospecifically blocked the effects of FK 33-824. The results indicate that the excitatory responses were predominantly mediated by mu-receptors.

Topics: Animals; Benzomorphans; Culture Techniques; Cyclazocine; D-Ala(2),MePhe(4),Met(0)-ol-enkephalin; Dose-Response Relationship, Drug; Electrophysiology; Endorphins; Enkephalin, Leucine-2-Alanine; Enkephalins; Ethylketocyclazocine; Hippocampus; Membrane Potentials; Morphinans; Rats; Receptors, Opioid; Receptors, Opioid, mu

Topics: Analgesics; Animals; Benzomorphans; Cyclazocine; Discrimination Learning; Discrimination, Psychological; Generalization, Stimulus; Male; Morphinans; Narcotics; Rats; Rats, Inbred Strains

Topics: Analgesia; Animals; Benzomorphans; Brain; Cyclazocine; Electric Stimulation; Ethylketocyclazocine; Guinea Pigs; Ileum; Macaca mulatta; Male; Mice; Morphinans; Morphine; Naloxone; Pentazocine; Rats; Receptors, Opioid; Receptors, Opioid, kappa; Vas Deferens