RESUMEN

Changes in the relative abundance of the G protein alpha subunits were observed during early mouse development Gs alpha was almost exclusively present as a large form (Gs-1) in prenatal brain. Postnatally with a substantial increase in Gpp[NH]p stimulated adenylyl cyclase activity, the small form (Gs.s) increased in amount while Gs-1 decreased. These results suggest that the Gs-s may be the more effective cyclase activator and that changes in alternative splicing are developmentally regulated. Gi1 and Go appeared before birth whereas Gi2 developed postnatally. Opiate stimulation of GTPase and inhibition of adenylyl cyclase were fully expressed prenatally.

Asunto(s)

Encéfalo/crecimiento & desarrollo , Proteínas de Unión al GTP/metabolismo , Adenilil Ciclasas/metabolismo , Animales , Western Blotting , Encéfalo/embriología , Encéfalo/metabolismo , AMP Cíclico/análisis , Etorfina/farmacología , Femenino , GTP Fosfohidrolasas/metabolismo , Expresión Génica/efectos de los fármacos , Expresión Génica/fisiología , Guanilil Imidodifosfato/farmacología , Cinética , Sustancias Macromoleculares , Masculino , Membranas/metabolismo , Ratones , Sinaptosomas/metabolismo

RESUMEN

Recently we reported the synthesis of the first enantiomeric pair of irreversible opioid ligands [(3S,4R)-(-)- and (3R,4S)-(+)-cis-4, SUPERFIT] and specific interaction of the latter with the delta receptor. Here we report another enantiomeric pair of irreversible opioid ligands, (+)-trans- and (-)-trans-3-methylfentanyl isothiocyanates [(3S,4S)-(+)-trans- and (3R,4R)-(-)-trans-4]. A single-crystal X-ray analysis of the 2,4,6-trinitrobenzenesulfonic acid salt of (+)-trans-3-methyl-N-phenyl-4-piperidinamine [(+)-trans-8] revealed it (and, therefore, 4) to have the trans configuration and the absolute configuration of (+)-trans-8 to be 3S,4S. The (+)-trans enantiomer of 4 was shown to be highly potent and about 10-fold more selective as an acylating agent than (-)-trans-4 for the higher affinity [3H]DADL (delta) binding site in rat brain membranes. In that assay, (+)-trans-4 and (+)-cis-4 were essentially equipotent as affinity ligands, and the levo enantiomers were considerably less potent. (+)-trans-4 was, thus, a potent, subtype-selective acylating agent for the delta opioid receptor in vitro. With membranes from NG108-15 neuroblastoma x glioma hybrid cells, containing only delta receptors, (+)-cis-4 was found to be a little more potent than (+)-trans-4. Similarly, (+)-cis-4 is the most effective inhibitor of adenylate cyclase in these membranes, (+)-trans-4 has weak activity, and the levo enantiomers are inactive. Only (+)-cis-4 was found to have antinociceptive activity in vivo.

Asunto(s)

Fentanilo/análogos & derivados , Receptores Opioides/metabolismo , Acilación , Inhibidores de Adenilato Ciclasa , Analgesia , Animales , Encéfalo/metabolismo , Membrana Celular/metabolismo , Fenómenos Químicos , Química , Encefalina Leucina/análogos & derivados , Encefalina Leucina/metabolismo , Encefalina Leucina/farmacología , Leucina Encefalina-2-Alanina , Encefalinas/metabolismo , Fentanilo/síntesis química , Fentanilo/metabolismo , Fentanilo/farmacología , Glioma/metabolismo , Células Híbridas , Indicadores y Reactivos , Ligandos , Masculino , Estructura Molecular , Neuroblastoma/metabolismo , Ratas , Ratas Endogámicas , Receptores Opioides delta , Estereoisomerismo , Relación Estructura-Actividad , Células Tumorales Cultivadas

RESUMEN

The mu and delta opiate receptors present in rat brain were measured independently during postnatal development. The numbers of delta receptors were almost undetectable at birth and increased substantially during the first few weeks, whereas the numbers of mu receptors remained relatively constant. Activation of either of these receptors caused inhibition of adenylate cyclase, but inhibition coupled to mu receptors was much smaller than that associated with delta receptors at all ages. Attempts to use pertussis toxin-catalyzed ADP-ribosylation as an assay for the GTP-binding proteins Ni and No were hampered by the development of an NADase with age. However, specific antibodies directed against the alpha subunits of Ni or No allowed separate quantitation of these transducer proteins. Both increased with age. No is present at levels at least 5-fold higher than Ni in the adult rat brain. The N proteins are in vast excess over receptors and as such are unlikely to be limiting factors in receptor function. The data further suggest that the number of opiate receptors present throughout neonatal development is in excess over that required for optimal function.

Asunto(s)

Encéfalo/crecimiento & desarrollo , Proteínas de Unión al GTP/metabolismo , Receptores Opioides/metabolismo , Envejecimiento , Animales , Encéfalo/metabolismo , Membrana Celular/metabolismo , Cinética , Ratas , Ratas Endogámicas , Receptores Opioides delta , Receptores Opioides mu

RESUMEN

N-Allyl-, N-(cyclopropylmethyl)-, and N-propyl-endo-ethenotetrahydronororipavines (N-substituted 6,14-endo-etheno-4,5-epoxy-3-hydroxy-6-methoxymorphinans) were synthesized with potential acylating or alkylating moieties at the C-7 position (isothiocyanato, (bromoacetyl)amino, and (methoxyfumaroyl)amino) and examined in vivo for their narcotic agonist and antagonist activities and for their ability to interact with opioid receptors in vitro. The N-(cyclopropylmethyl)-substituted compounds were found to have the highest affinity for opioid receptors among these N-substituted compounds, although all of them were found to be reasonably potent narcotic antagonists in the mouse tail flick vs. morphine assay. Their in vivo potency ranged from 1/8 to 4 times that of nalorphine on intravenous injection in mice. Rat brain membrane binding studies indicated that the compounds interacted with opioid receptors with potencies that ranged from 0.5 times that of morphine (8c, 9c, and 10c) to 0.017 that of morphine (8b). Among the compounds studied here, only the previously reported isothiocyanato compound (10c) and (methoxyfumaroyl)amino compound (8c) interacted irreversibly and selectively with mu or delta opioid receptors, respectively, in assays using NG108-15 neuroblastoma-glioma hybrid cells and/or in a rat brain membrane preparation. Both 8c and 10c were found to interact irreversibly, to a limited extent, with kappa opioid sites in rat brain membranes in which the mu and delta opioid receptors were depleted by interaction with the mu-selective irreversible ligand BIT and the delta-selective irreversible ligand FIT. Neither compound showed irreversible actions in the electrically stimulated mouse vas deferens preparation.

Asunto(s)

Encefalina Leucina/análogos & derivados , Isotiocianatos , Antagonistas de Narcóticos/síntesis química , Tebaína/análogos & derivados , Animales , Encéfalo/metabolismo , Fentanilo/análogos & derivados , Fentanilo/farmacología , Técnicas In Vitro , Ligandos , Masculino , Ratones , Ratones Endogámicos ICR , Antagonistas de Narcóticos/metabolismo , Antagonistas de Narcóticos/farmacología , Oligopéptidos/farmacología , Ratas , Receptores Opioides/efectos de los fármacos , Receptores Opioides delta , Receptores Opioides mu , Relación Estructura-Actividad , Tiocianatos/farmacología

RESUMEN

The first enantiomeric pair of irreversible opioid ligands [(+)- and (-)-4] were synthesized in greater than 99.6% optical purity as determined by HPLC analysis of diastereoisomeric derivatives of the intermediate 3-methyl-N-phenyl-4-piperidinamine enantiomers. Single-crystal X-ray analysis of the (R,R)-L-(+)-tartaric acid salt of (-)-9 revealed the absolute configuration to be 3S,4R. The absolute configuration of (-)-3 [cis-(-)-3-methylfentanyl] and (-)-4 derived from (-)-9 is thus 3S,4R and that of (+)-3 and (+)-4 is 3R,4S. The (+) enantiomer of 4 (SUPERFIT) was shown to be highly potent and specific for acylation of delta opioid receptors (to the exclusion of mu) in rat brain membranes like its achiral prototype FIT and was about 10 times as potent as the latter in this assay. The (+)-4 was about 5 times as potent as FIT in acylation of delta receptors in NG108-15 neuroblastoma X glioma hybrid cells and about 50 times as potent as its enantiomer. Both FIT and (+)-4 behaved as partial agonists in inhibition of delta receptor coupled adenylate cyclase in NG108-15 membranes and (+)-4 was 5-10 times more potent than FIT and about 100 times more potent than its enantiomer in this assay. Dibromination of amine 12, catalytic exchange of bromine with tritium gas, and reaction of the labeled amine with thiophosgene afforded [3H]-(+)-4 with a specific activity of 13 Ci/mmol. Previous experiments indicated (+)-4 acylates the same 58 000-dalton glycoprotein previously shown to be acylated by FIT but with less nonspecific labeling. In view of the high potency and specificity of (+)-4 and the availability of its enantiomer, it seems likely that these compounds will prove to be valuable tools for study of the opioid receptor complex.

Asunto(s)

Fentanilo/análogos & derivados , Isotiocianatos , Receptores Opioides/metabolismo , Acilación , Inhibidores de Adenilato Ciclasa , Analgésicos/farmacología , Animales , Encéfalo/metabolismo , Encefalina Metionina/análogos & derivados , Encefalina Metionina/metabolismo , Fentanilo/síntesis química , Fentanilo/farmacología , Técnicas In Vitro , Ligandos , Ratones , Conformación Molecular , Ratas , Receptores Opioides/análisis , Receptores Opioides delta , Receptores Opioides mu , Estereoisomerismo , Tritio , Difracción de Rayos X

RESUMEN

The addition of bradykinin to NG108-15 cells results in a transient hyperpolarization followed by prolonged cell depolarization. Injection of inositol 1,4,5-trisphosphate or Ca2+ into the cytoplasm of NG108-15 cells also elicits cell hyperpolarization followed by depolarization. Tetraethylammonium ions inhibit the hyperpolarizing response of cells to bradykinin or inositol 1,4,5-trisphosphate. Thus, the hyperpolarizing phase of the cell response may be due to inositol 1,4,5-trisphosphate-dependent release of stored Ca2+ into the cytoplasm, which activates Ca2+-dependent K+ channels. The depolarizing phase of the cell response to bradykinin is due largely to inhibition of M channels, thereby decreasing the rate of K+ efflux from cells and, to a lesser extent, to activation of Ca2+-dependent ion channels and Ca2+ channels. In contrast, injection of inositol 1,4,5-trisphosphate or Ca2+ into the cytosol did not alter M channel activity. Incubation of NG108-15 cells with pertussis toxin inhibits bradykinin-dependent cell hyperpolarization and depolarization. Bradykinin stimulates low Km GTPase activity and inhibits adenylate cyclase in NG108-15 membrane preparations but not in membranes prepared from cells treated with pertussis toxin. Reconstitution of NG108-15 membranes from cells treated with pertussis toxin with nanomolar concentrations of a mixture of highly purified No and Ni [guanine nucleotide-binding proteins that have no known function (No) or inhibit adenylate cyclase (Ni)] restores bradykinin-dependent activation of GTPase and inhibition of adenylate cyclase. These results show that [bradykinin . receptor] complexes interact with No or Ni and suggest that No and/or Ni mediate the transduction of signals from bradykinin receptors to phospholipase C and adenylate cyclase.

Asunto(s)

Inhibidores de Adenilato Ciclasa , Bradiquinina/farmacología , Proteínas de Unión al GTP/fisiología , Glioma/fisiopatología , Fosfatos de Inositol/farmacología , Potenciales de la Membrana/efectos de los fármacos , Neuroblastoma/fisiopatología , Fosfatos de Azúcar/farmacología , Toxina de Adenilato Ciclasa , Calcio/farmacología , Línea Celular , Diglicéridos/farmacología , Humanos , Células Híbridas/fisiología , Inositol 1,4,5-Trifosfato , Toxina del Pertussis , Acetato de Tetradecanoilforbol/farmacología , Tetraetilamonio , Compuestos de Tetraetilamonio/farmacología , Factores de Virulencia de Bordetella/farmacología

Asunto(s)

Proteínas de Unión al GTP/fisiología , Receptores Opioides/fisiología , Animales , Química Encefálica , Bovinos , Toxina del Cólera/farmacología , GTP Fosfohidrolasas/antagonistas & inhibidores , GTP Fosfohidrolasas/metabolismo , Ratones , Receptores Opioides delta , Factores de Virulencia de Bordetella/farmacología

RESUMEN

Syntheses of affinity reagents for opiate receptors based on the fentanyl, endo-ethenotetrahydrooripavine, and etonitazene carbon-nitrogen skeletons are described. The isothiocyanate, bromoacetamido, and methylfumaramido alkylating functions were employed in these compounds, some of which had previously been shown to be mu specific (12, BIT) and delta specific (8, FIT and 19, FAO) in vitro. Antinociceptive activity of the title compounds was determined in the mouse hot-plate test, which revealed that certain compounds in each class showed morphine-like activity. The binding EC50 values against [3H]Dalamid for opiate receptors in NG108-15 (delta receptors) and rat brain membranes (mu + delta receptors) are also reported. With this type of experiment, it was possible to independently measure the apparent affinity of the etonitazene congeners 12-14 for the mu and delta receptors.

Asunto(s)

Ligandos/metabolismo , Antagonistas de Narcóticos/síntesis química , Receptores Opioides/metabolismo , Animales , Bencimidazoles/análogos & derivados , Bencimidazoles/metabolismo , Bencimidazoles/farmacología , Línea Celular , Membrana Celular/metabolismo , Fenómenos Químicos , Química , Evaluación Preclínica de Medicamentos , Fentanilo/análogos & derivados , Fentanilo/metabolismo , Fentanilo/farmacología , Glioma/metabolismo , Células Híbridas/metabolismo , Indicadores y Reactivos , Ligandos/farmacología , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Ratones , Neuroblastoma/metabolismo , Ratas , Receptores Opioides/efectos de los fármacos , Receptores Opioides delta , Receptores Opioides mu , Relación Estructura-Actividad , Tebaína/análogos & derivados , Tebaína/metabolismo , Tebaína/farmacología

RESUMEN

A p-isothiocyanatophenylacetylamino moiety at the C7alpha position on an endoethenooripavine (NIH 10358) was found to irreversibly bind to both the mu and delta opioid receptors in rat brain membrane preparations. However, the same endoethenooripavine nucleus with a p-methylfumaroylaminophenylacetylamino moiety at that position (NIH 10364) binds specifically and irreversibly to only mu opioid receptors. Hypotheses are being developed concerning receptor subclass alkylation specificity. For example, it has now been found that the delta receptor irreversible agonist, FIT, can interact with both mu and delta receptors in a reversible manner.

Asunto(s)

Fentanilo/análogos & derivados , Isotiocianatos , Derivados de la Morfina , Receptores Opioides/metabolismo , Alquilantes/metabolismo , Animales , Bencimidazoles/metabolismo , Unión Competitiva , Encéfalo/metabolismo , Fentanilo/metabolismo , Técnicas In Vitro , Membranas/metabolismo , Ratas , Receptores Opioides delta , Receptores Opioides mu , Tebaína/análogos & derivados , Tebaína/metabolismo

RESUMEN

Exorphins, peptides with opioid activity, have previously been isolated from pepsin hydrolysates of alpha-casein [Zioudrou, C., Streaty, R. A., & Klee, W. A. (1979) J. Biol. Chem. 254, 2446-2449]. Analysis of these peptides shows that they correspond to the sequences 90-96, Arg-Tyr-Leu-Gly-Tyr-Leu-Glu, and 90-95, Arg-Tyr-Leu-Gly-Tyr-Leu, of alpha-casein. These peptides, as well as two of their analogues Tyr-Leu-Gly-Tyr-Leu-Glu (91-96) and Tyr-Leu-Gly-Tyr-Leu (91-95), have now been synthesized and characterized. Their opioid activity was examined by three different bioassays: (a) displacement of D-2-alanyl[tyrosyl-3,5-3H]enkephalin-(5-L-methioninamide) and [3H]dihydromorphine from rat brain membranes; (b) naloxone-reversible inhibition of adenylate cyclase in homogenates of neuroblastoma x glioma hybrid cells; (c) naloxone-reversible inhibition of electrically stimulated contractions of the mouse vas deferens. The synthetic peptide of sequence 90-96 was the most potent opioid in all three bioassays and its potency was similar to that of the isolated alpha-casein exorphins. The synthetic peptides were totally resistant to hydrolysis by trypsin and homogenates of rat brain membranes, but were partially inactivated by chymotrypsin and subtilisin. The difference in opioid activity of alpha-casein exorphins may be related to differences in conformational flexibility observed by NMR spectroscopy.

Asunto(s)

Caseínas , Péptidos/farmacología , Inhibidores de Adenilato Ciclasa , Secuencia de Aminoácidos , Animales , Bioensayo , Encéfalo/metabolismo , Línea Celular , Hidrólisis , Indicadores y Reactivos , Cinética , Espectroscopía de Resonancia Magnética , Pepsina A , Péptidos/síntesis química , Péptidos/aislamiento & purificación , Ratas , Receptores Opioides/metabolismo , Membranas Sinápticas/metabolismo

RESUMEN

Photolabile derivatives of D-Ala2-Leu5-enkephalin were prepared by synthetic procedures in which a 2-nitro-4-azidophenyl group is linked to the terminal carboxyl group of the enkephalin by means of an ethylenediamine or ethylenediamine beta-alanine spacer. These peptides bind to opiate receptors with nanomolar affinities and inhibit electrically stimulated contractions of the mouse vas deferens and adenylate cyclase activity of NG108-15 neuroblastoma x glioma hybrid cell membranes. Both inhibitions are reversed by the opiate antagonist naloxone. Photolysis of the ligands bound to rat brain membranes results in the loss of approximately 50% of the receptor sites. This decrease in receptor number is blocked by naloxone and requires light. A photolabile [3H]enkephalin derivative labels an equivalent number of sites under similar irradiation conditions.

Asunto(s)

Encefalina Leucina/análogos & derivados , Receptores Opioides/metabolismo , Animales , Bioensayo , Encéfalo/metabolismo , Encefalina Leucina/metabolismo , Encefalina Leucina/farmacología , Leucina Encefalina-2-Alanina , Masculino , Ratones , Contracción Muscular/efectos de los fármacos , Fotólisis , Ratas , Conducto Deferente/efectos de los fármacos

RESUMEN

Alkylating agents that display strong selectivity for opiate receptor types delta or mu were prepared by appropriate modification of the structures of the strong analgesics fentanyl, etonitazene, and endoethenotetrahydrooripavine. The availability of these substances should facilitate studies of the structural basis of receptor specificity and of the physiologic roles of these receptors.

Asunto(s)

Isotiocianatos , Receptores Opioides/metabolismo , Alquilación , Animales , Bencimidazoles/análogos & derivados , Bencimidazoles/metabolismo , Encéfalo/fisiología , Células Cultivadas , Fenómenos Químicos , Química , Encefalina Metionina/análogos & derivados , Encefalina Metionina/metabolismo , Fentanilo/análogos & derivados , Fentanilo/metabolismo , Ligandos , Ratas , Receptores Opioides/fisiología , Tebaína/análogos & derivados , Tebaína/farmacología

RESUMEN

Nine new compounds have been synthesized as potential affinity ligands for specific opioid receptors. The biochemical properties of three of these compounds were examined in detail and one of them, N-cyclopropylmethyl-7 alpha-methylfumaroylamido-6, 14-endoethenotetrahydronororipavine (NIH 10236), was found to be a potent irreversible ligand for the delta opioid receptor. It had the properties of a narcotic antagonist, as determined by its effect on adenylate cyclase activity of NG108-15 neuroblastoma-glioma cell homogenates. It is, thus, the first delta specific alkylating ligand known which is a narcotic antagonist. A second compound, the N-cyclopropylmethyl-7 alpha-isothiocyanato-6, 14-endoethenotetrahydronororipavine (NIH 10235) was found to be a mu specific alkylating ligand in brain and a reversible antagonist in the NG108-15 cells.

Asunto(s)

Morfinanos/farmacología , Receptores Opioides/metabolismo , Tebaína/análogos & derivados , Animales , Unión Competitiva , Encéfalo/metabolismo , Línea Celular , Membrana Celular/metabolismo , Glioma/metabolismo , Células Híbridas/metabolismo , Ligandos , Ratones , Neuroblastoma/metabolismo , Ratas , Receptores Opioides/efectos de los fármacos

RESUMEN

Opiates and opioid peptides inhibit adenylate cyclase and stimulate specific low Km GTPase activity in membranes from neuroblastoma x glioma NG108-15 hybrid cells. The effects of opiate agonists on both enzymes are mediated by high affinity stereospecific receptors and require Mg2+, GTP, and Na+. In the presence of Mg2+, Na+ inhibits basal GTPase activity; opiates stimulate GTP hydrolysis by antagonizing the Na+-induced inhibition. Activation of GTPase leads, in turn, to inactivation of GTP-stimulated adenylate cyclase activity. The intrinsic activities (or efficacies) of a series of opiates are identical for stimulation of GTPase and inhibition of adenylate cyclase. These results provide a mechanism for the dual requirement for Na+ and GTP in the inhibitory coupling of opiate receptors to the adenylate cyclase system in these cells and may be of general significance to the action of other inhibitory hormones.

Asunto(s)

Inhibidores de Adenilato Ciclasa , GTP Fosfohidrolasas/metabolismo , Guanosina Trifosfato/farmacología , Narcóticos/farmacología , Monoéster Fosfórico Hidrolasas/metabolismo , Sodio/farmacología , Animales , Cationes Monovalentes , Línea Celular , Glioma/enzimología , Células Híbridas/enzimología , Cinética , Magnesio/farmacología , Cloruro de Magnesio , Ratones , Neuroblastoma/enzimología , Ratas

RESUMEN

The 2-nitro-4-azidophenyl(NAP)-D-Ala2-Leu5-Enkephalin derivatives: Try-D-Ala-Gly-Phe-Leu CONCH2CH2NH-NAP (E-NAP-EDA) and Try-D-Ala-Gly-Phe-Leu CONCH2CH2NH-COCH2CH2NHNAP(E-NAP- -Ala-EDA) were synthesized by conventional peptide methods. Their structure was determined by amino acid analysis, ultra violet, visible and infra red spectroscopy. Both peptides were shown a) to bind with high affinity to the opiate receptors of rat brain membranes and b) to inhibit strongly the contractions of electrically stimulated vas deferens and the adenyl cyclase of the NG 108-15 cell membranes. These effects were reversed by the antagonist naloxone. Photoloysis of the rat brain membranes-(E-NAP- -Ala-EDA) complex caused a 20-30% inactivation of the opiate receptors. Inactivation was prevented when the complex was irradiated in the presence of naloxone. The radio-labeled derivatives of these enkephalin analogs may prove useful photochemical labels of the opiate receptor.

Asunto(s)

Ligandos/síntesis química , Receptores Opioides/metabolismo , Animales , Encéfalo/metabolismo , Endorfinas/metabolismo , Encefalina Leucina/análogos & derivados , Leucina Encefalina-2-Alanina , Fotólisis , Ratas

Asunto(s)

Adenilil Ciclasas/metabolismo , Narcóticos/farmacología , Receptores Opioides/metabolismo , Animales , Línea Celular , Sistema Libre de Células , Glioma/enzimología , Células Híbridas/enzimología , Cinética , Ratones , Neuroblastoma/enzimología , Ratas

RESUMEN

Receptors that reversibly bind opiates and opioid peptides have been solubilized from brain and neuroblastoma-glioma hybrid cell NG108-15 membranes. Active receptors are specifically solubilized with a new type of detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, which is a zwitterionic derivative of cholic acid. The solubilized receptor complexes behave as large molecules with a Stokes radius of 70 A and contain protein as an essential constituent.

Asunto(s)

Química Encefálica , Neuronas/análisis , Receptores Opioides/aislamiento & purificación , Animales , Bovinos , Línea Celular , Detergentes , Etilmaleimida/farmacología , Ratas , Receptores Opioides/efectos de los fármacos , Solubilidad , Tripsina/metabolismo

RESUMEN

Peptides with opioid activity are found in pepsin hydrolysates of wheat gluten and alpha-casein. The opioid activity of these peptides was demonstrated by use of the following bioassays: 1) naloxone-reversible inhibition of adenylate cyclase in homogenates of neuroblastoma X-glioma hybrid cells; 2) naloxone-reversible inhibition of electrically stimulated contractions of the mouse vas deferens; 3) displacement of [3H]dihydromorphine and [3H-Tyr, dAla2]met-enkephalin amide from rat brain membranes. Substances which stimulate adenylate cyclase and increase the contractions of the mouse vas deferens but do not bind to opiate receptors are also isolated from gluten hydrolysates. It is suggested that peptides derived from some food proteins may be of physiological importance.

Asunto(s)

Narcóticos , Péptidos , Adenilil Ciclasas/metabolismo , Animales , Bioensayo , Caseínas , Proteínas en la Dieta , Glútenes , Cinética , Masculino , Narcóticos/farmacología , Pepsina A , Péptidos/farmacología

RESUMEN

Several 3-deoxy opioids and 3,6-dideoxydihydromorphine was synthesized to ascertain the effect of the phenolic hydroxyl group on antinociceptive potency and receptor binding affinity. Catalytic reduction of the 3-tetrazolyl ether derivatives of dihydromorphine provided the entry into the 3-deoxydihydro series. The prototype, 3-deoxymorphine, was prepared by lithium aluminum hydride reduction of 3-deoxy-N-carbethoxymorphinone, obtained via its 7-(phenylseleno) derivative. 3-Deoxydihydromorphinone and 3,6-dideoxydihydromorphine were found to be about as potent as, or more potent than, morphine in standard antiociceptive assays. Each of them, however, was less potent than the comparable 3-hydroxy analogue, and their binding affinity to the opiate receptor was substantially decreased. The epoxy ring in 3.6-dideoxydihydromorphine was found to increase the antinociceptive potency of the compound.

Asunto(s)

Analgésicos , Derivados de la Morfina/farmacología , Receptores Opioides/efectos de los fármacos , Animales , Fenómenos Químicos , Química , Ratones , Derivados de la Morfina/síntesis química , Relación Estructura-Actividad

RESUMEN

The pharmacological effect of the N-(beta-cyanoethyl) moiety is dependent on the opioid on which it is substituted. It caused a large increase in antinociceptive potency, in (--)-3-hydroxymorphinan and (--)-normetazocine, as compared with the N-methyl opioid. These cyanoethyl compounds do not substitute for morphine in morphine-dependent monkeys. This moiety also appears to greatly increase the ability of the opiate receptor to differentiate enantiomers. An ca. 100,000-fold difference in binding was noted between the epimeric N-(beta-cyanoethyl)-3-hydroxymorphinans and the normetazocines. The levo enantiomers have little acute toxicity and showed excellent therapeutic ratios. In contrast, the N-(beta-cyanoethyl) moiety on normorphine, norcodeine, and noroxymorphone did not appear to improve their pharmacological properties. Homologous N-cyanoalkyl opioids were less potent antinociceptives.

Asunto(s)

Narcóticos/farmacología , Alquilación , Analgésicos , Animales , Fenómenos Químicos , Química , Haplorrinos , Dosificación Letal Mediana , Macaca mulatta , Ratones , Narcóticos/síntesis química , Narcóticos/metabolismo , Ratas , Receptores Opioides/efectos de los fármacos , Relación Estructura-Actividad