RESUMEN
Improgan, a non-opioid, antinociceptive drug, activates descending analgesic circuits following brain administration, but the improgan receptor remains unidentified. Since biotinylation of drugs can enhance drug potency or facilitate discovery of new drug targets, a biotinylated congener of improgan (CC44) and several related compounds were synthesized and tested for antinociceptive activity. In rats and mice, intracerebroventricular (i.c.v.) administration of CC44 produced dose-dependent reductions in thermal nociceptive (tail flick and hot plate) responses, with 5-fold greater potency than improgan. CC44 also robustly attenuated mechanical (tail pinch) nociception in normal rats and mechanical allodynia in a spinal nerve ligation model of neuropathic pain. Similar to the effects of improgan, CC44 antinociception was reversed by the GABAA agonist muscimol (consistent with activation of analgesic circuits), and was resistant to the opioid antagonist naltrexone (implying a non-opioid mechanism). Also like improgan, CC44 produced thermal antinociception when microinjected into the rostral ventromedial medulla (RVM). Unlike improgan, CC44 (i.c.v.) produced antinociception which was resistant to antagonism by the cannabinoid CB1 antagonist/inverse agonist rimonabant. CC44 was inactive in mice following systemic administration, indicating that CC44 does not penetrate the brain. Preliminary findings with other CC44 congeners suggest that the heteroaromatic nucleus (imidazole), but not the biotin moiety, is required for CC44's antinociceptive activity. These findings demonstrate that CC44 is a potent analgesic compound with many improgan-like characteristics. Since powerful techniques are available to characterize and identify the binding partners for biotin-containing ligands, CC44 may be useful in searching for new receptors for analgesic drugs.
Asunto(s)
Analgésicos/química , Analgésicos/farmacología , Biotinilación , Cimetidina/análogos & derivados , Analgésicos/metabolismo , Analgésicos/uso terapéutico , Animales , Avidina/metabolismo , Cimetidina/química , Cimetidina/metabolismo , Cimetidina/farmacología , Cimetidina/uso terapéutico , Hiperalgesia/tratamiento farmacológico , Masculino , Bulbo Raquídeo/patología , Ratones , Neuronas/efectos de los fármacos , Neuronas/patología , Ratas , Estreptavidina/metabolismoRESUMEN
Derivatives of the lead compound N-BPE-8-CAC (1) where each CH of the biphenyl group was individually replaced by N were prepared in hopes of identifying high affinity ligands with improved aqueous solubility. Compared to 1, binding affinities of the five possible pyridinyl derivatives for the µ opioid receptor were between threefold lower to fivefold higher with the Ki of the most potent compound being 0.064 nM. Docking of 8-CAC (2) into the unliganded binding site of the mouse µ opioid receptor (pdb: 4DKL) revealed that 8-CAC and ß-FNA (from 4DKL) make nearly identical interactions with the receptor. However, for 1 and the new pyridinyl derivatives 4-8, binding is not tolerated in the 8-CAC binding mode due to the steric constraints of the large N-substituents. Either an alternative binding mode or rearrangement of the protein to accommodate these modifications may account for their high binding affinity.
Asunto(s)
Compuestos de Bifenilo/farmacología , Ciclopropanos/farmacología , Piridinas/farmacología , Receptores Opioides/química , Animales , Compuestos de Bifenilo/síntesis química , Compuestos de Bifenilo/química , Ciclopropanos/síntesis química , Ciclopropanos/química , Relación Dosis-Respuesta a Droga , Humanos , Ligandos , Ratones , Modelos Moleculares , Estructura Molecular , Piridinas/síntesis química , Piridinas/química , Receptores Opioides/metabolismo , Relación Estructura-ActividadRESUMEN
N-[2-(4'-methoxy[1,1'-biphenyl]-4-yl)ethyl]-8-CAC (1) is a high affinity (K(i)=0.084 nM) ligand for the µ opioid receptor and served as the lead compound for this study. Analogues of 1 were made in hopes of identifying an SAR within a series of oxygenated (distal) phenyl derivatives. A number of new analogues were made having single-digit pM affinity for the µ receptor. The most potent was the 3',4'-methylenedioxy analogue 18 (K(i)=1.6 pM).
Asunto(s)
Ciclazocina/análogos & derivados , Oxígeno/química , Receptores Opioides mu/agonistas , Receptores Opioides mu/antagonistas & inhibidores , Ciclazocina/síntesis química , Ciclazocina/química , Ciclazocina/farmacología , Relación Dosis-Respuesta a Droga , Humanos , Ligandos , Estructura Molecular , Relación Estructura-ActividadRESUMEN
Conventional chemotherapy not only kills tumor cells but also changes gene expression in treatment-damaged tissues, inducing production of multiple tumor-supporting secreted factors. This secretory phenotype was found here to be mediated in part by a damage-inducible cell-cycle inhibitor p21 (CDKN1A). We developed small-molecule compounds that inhibit damage-induced transcription downstream of p21. These compounds were identified as selective inhibitors of a transcription-regulating kinase CDK8 and its isoform CDK19. Remarkably, p21 was found to bind to CDK8 and stimulate its kinase activity. p21 and CDK8 also cooperate in the formation of internucleolar bodies, where both proteins accumulate. A CDK8 inhibitor suppresses damage-induced tumor-promoting paracrine activities of tumor cells and normal fibroblasts and reverses the increase in tumor engraftment and serum mitogenic activity in mice pretreated with a chemotherapeutic drug. The inhibitor also increases the efficacy of chemotherapy against xenografts formed by tumor cell/fibroblast mixtures. Microarray data analysis revealed striking correlations between CDK8 expression and poor survival in breast and ovarian cancers. CDK8 inhibition offers a promising approach to increasing the efficacy of cancer chemotherapy.
Asunto(s)
Antineoplásicos/farmacología , Quinasa 8 Dependiente de Ciclina/fisiología , Regulación Neoplásica de la Expresión Génica , Neoplasias/tratamiento farmacológico , Animales , Línea Celular Tumoral , Nucléolo Celular/metabolismo , Senescencia Celular , Quinasa 8 Dependiente de Ciclina/metabolismo , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Genómica , Humanos , Ratones , Ratones Endogámicos C57BL , Trasplante de Neoplasias , Neoplasias/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , Regiones Promotoras Genéticas , Proteínas Quinasas Asociadas a Fase-S/metabolismo , Transcripción Genética , Resultado del TratamientoRESUMEN
Cytochrome P450 (CYP)-mediated epoxidation of arachidonic acid (AA) contributes to important biological functions, including the pain-relieving responses produced by analgesic drugs. However, the relevant epoxygenase(s) remain unidentified. Presently, we describe the tissue distribution, high-throughput assay, and pharmacological characteristics of the rat epoxygenase CYP2C24. Following cloning from male rat liver, recombinant baculovirus containing the C-terminal His-tagged cDNA was constructed and used to express the protein in Spodoptera frugiperda (Sf9) cells. Enzymatic activity was detected with membranes, NADPH regenerating system and CYP reductase, and optimized for high throughput screening by use of the Vivid Blue© BOMCC fluorescence substrate. Quantitative real-time PCR identified CYP2C24 m-RNA in liver, kidney, heart, lung, gonad and brain. Screening of CYP2C24 activity against a panel of inhibitors showed a very strong correlation with activity against the human homologue CYP2C19. In agreement with recent findings on CYP2C19, the epoxygenase blockers PPOH and MS-PPOH inhibited CYP2C24 only weakly, confirming that these drugs are not universal epoxygenase inhibitors. Finally, comparisons of the CYP2C24 inhibitor profile with anti-analgesic activity suggests that this isoform does not contribute to brain analgesic drug action. The present methods and pharmacological data will aid in study of the biological significance of this CYP isoform.
RESUMEN
The search for the mechanism of action of improgan (a nonopioid analgesic) led to the recent discovery of CC12, a compound that blocks improgan antinociception. Because CC12 is a cytochrome P450 inhibitor, and brain P450 mechanisms were recently shown to be required in opioid analgesic signaling, pharmacological and transgenic studies were performed in rodents to test the hypothesis that improgan antinociception requires brain P450 epoxygenase activity. Intracerebroventricular (i.c.v.) administration of the P450 inhibitors miconazole and fluconazole, and the arachidonic acid (AA) epoxygenase inhibitor N-methylsulfonyl-6-(2-propargyloxyphenyl)hexanamide (MS-PPOH) potently inhibited improgan antinociception in rats at doses that were inactive alone. MW06-25, a new P450 inhibitor that combines chemical features of CC12 and miconazole, also potently blocked improgan antinociception. Although miconazole and CC12 were weakly active at opioid and histamine H(3) receptors, MW06-25 showed no activity at these sites, yet retained potent P450-inhibiting properties. The P450 hypothesis was also tested in Cpr(low) mice, a viable knock-in model with dramatically reduced brain P450 activity. Improgan (145 nmol, i.c.v.) antinociception was reduced by 37% to 59% in Cpr(low) mice, as compared with control mice. Moreover, CC12 pretreatment (200 nmol, i.c.v.) abolished improgan action (70% to 91%) in control mice, but had no significant effect in Cpr(low) mice. Thus, improgan's activation of bulbospinal nonopioid analgesic circuits requires brain P450 epoxygenase activity. A model is proposed in which (1) improgan activates an unknown receptor to trigger downstream P450 activity, and (2) brainstem epoxygenase activity is a point of convergence for opioid and nonopioid analgesic signaling.
Asunto(s)
Analgésicos no Narcóticos/farmacología , Encéfalo/efectos de los fármacos , Cimetidina/análogos & derivados , Sistema Enzimático del Citocromo P-450/metabolismo , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Inhibidores de 14 alfa Desmetilasa/farmacología , Amidas/farmacología , Analgésicos Opioides/farmacocinética , Animales , Encéfalo/metabolismo , Línea Celular Transformada , Cimetidina/farmacología , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/farmacología , Humanos , Imidazoles/farmacología , Inyecciones Intraventriculares/métodos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Miconazol/farmacología , NADPH-Ferrihemoproteína Reductasa/deficiencia , Naltrexona/análogos & derivados , Naltrexona/farmacocinética , Antagonistas de Narcóticos/farmacocinética , Dimensión del Dolor/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Tiempo de Reacción/efectos de los fármacos , Receptores Histamínicos H3/metabolismo , Sulfuros/farmacología , Factores de Tiempo , Tritio/farmacocinéticaRESUMEN
To assess the importance of brain cytochrome P450 (P450) activity in mu opioid analgesic action, we generated a mutant mouse with brain neuron-specific reductions in P450 activity; these mice showed highly attenuated morphine antinociception compared with controls. Pharmacological inhibition of brain P450 arachidonate epoxygenases also blocked morphine antinociception in mice and rats. Our findings indicate that a neuronal P450 epoxygenase mediates the pain-relieving properties of morphine.
Asunto(s)
Analgésicos Opioides/farmacología , Encéfalo/efectos de los fármacos , Sistema Enzimático del Citocromo P-450/efectos de los fármacos , Neuronas/efectos de los fármacos , Dolor/tratamiento farmacológico , Receptores Opioides mu/metabolismo , Analgésicos Opioides/administración & dosificación , Animales , Encéfalo/enzimología , Encéfalo/metabolismo , Citocromo P-450 CYP2J2 , Inhibidores Enzimáticos del Citocromo P-450 , Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/farmacología , Femenino , Masculino , Ratones , Ratones Transgénicos , Morfina/administración & dosificación , Morfina/farmacología , Vías Nerviosas/efectos de los fármacos , Vías Nerviosas/enzimología , Vías Nerviosas/metabolismo , Neuronas/enzimología , Neuronas/metabolismo , Dolor/enzimología , Dolor/metabolismo , Ratas , Ratas Sprague-Dawley , Transducción de Señal , Factores de TiempoRESUMEN
A series of novel high affinity opioid receptor ligands have been made whereby the phenolic-OH group of nalbuphine, naltrexone methiodide, 6-desoxonaltrexone, hydromorphone and naltrindole was replaced by a carboxamido group and the furan ring was opened to the corresponding 4-OH derivatives. These furan ring 'open' derivatives display very high affinity for mu and kappa receptors and much less affinity for delta. The observation that these target compounds have much higher receptor affinity than the corresponding ring 'closed' carboxamides significantly strengthens our underlying pharmacophore hypothesis concerning the bioactive conformation of the carboxamide group.
Asunto(s)
Analgésicos Opioides/síntesis química , Antagonistas de Narcóticos/síntesis química , Receptores Opioides/metabolismo , Analgésicos Opioides/metabolismo , Cristalografía por Rayos X , Furanos/síntesis química , Furanos/metabolismo , Humanos , Ligandos , Naltrexona/síntesis química , Naltrexona/metabolismo , Antagonistas de Narcóticos/metabolismo , Unión Proteica , Receptores Opioides/agonistasRESUMEN
A series of 15 novel opioid derivatives were made where the prototypic phenolic-OH group of traditional opioids was replaced by a carboxamido (CONH(2)) group. For 2,6-methano-3-benzazocines and morphinans similar or, in a few instances, enhanced affinity for mu, delta and kappa opioid receptors was observed when the OH-->CONH(2) switch was applied. For 4,5alpha-epoxymorphinans, binding affinities for the corresponding carboxamide derivatives were much lower than the OH partner consistent with our pharmacophore hypothesis concerning carboxamide bioactive conformation. The active metabolite of tramadol and its carboxamide counterpart had comparable affinities for the three receptors.
Asunto(s)
Analgésicos Opioides/síntesis química , Receptores Opioides/metabolismo , Amidas , Analgésicos Opioides/metabolismo , Analgésicos Opioides/farmacología , Humanos , Unión Proteica , Relación Estructura-Actividad , Tramadol/metabolismoRESUMEN
A series of 7,8- and 8,9-fused triazole and imidazole analogues of cyclazocine have been made and characterized in opioid receptor binding and [(35)S]GTPgammaS assays. Target compounds were designed to explore the SAR surrounding our lead molecule for this study, namely the 8,9-fused pyrrolo analogue 2 of cyclazocine. Compared to 2, many of the new compounds in this study displayed very high affinity for opioid receptors.
Asunto(s)
Ciclazocina/síntesis química , Ciclazocina/farmacología , Receptores Opioides/efectos de los fármacos , Animales , Células CHO , Cricetinae , Cricetulus , Ciclazocina/análogos & derivados , Ciclazocina/metabolismo , Ciclización , Guanosina 5'-O-(3-Tiotrifosfato)/metabolismo , Ensayo de Unión Radioligante , Receptores Opioides/metabolismo , Relación Estructura-Actividad , Radioisótopos de AzufreRESUMEN
A series of 7,8- and 8,9-fused pyrimidinone, aminopyrimidine and pyridone derivatives of 8-carboxamidocyclazocine (8-CAC) have been prepared and evaluated in opioid receptor binding assays. Targets were designed to corroborate a pharmacophore hypothesis regarding the bioactive conformation of the carboxamide of 8-CAC. In addition to the results from this study strongly supporting this pharmacophore hypothesis, a number of novel compounds with high affinity to opioid receptors have been identified.
Asunto(s)
Azocinas/farmacología , Ciclazocina/análogos & derivados , Antagonistas de Narcóticos , Receptores Opioides/agonistas , Animales , Azocinas/síntesis química , Azocinas/química , Unión Competitiva/efectos de los fármacos , Células CHO , Cricetinae , Cricetulus , Ciclazocina/síntesis química , Ciclazocina/química , Ciclazocina/farmacología , Diseño de Fármacos , Conformación Molecular , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
A series of aryl-containing N-monosubstituted analogues of the lead compound 8-[N-((4'-phenyl)-phenethyl)]-carboxamidocyclazocine were synthesized and evaluated to probe a putative hydrophobic binding pocket of opioid receptors. Very high binding affinity to the mu opioid receptor was achieved though the N-(2-(4'-methoxybiphenyl-4-yl)ethyl) analogue of 8-CAC. High binding affinity to mu and very high binding affinity to kappa opioid receptors was observed for the N-(3-bromophenethyl) analogue of 8-CAC. High binding affinity to all three opioid receptors were observed for the N-(2-naphthylethyl) analogue of 8-CAC.
Asunto(s)
Ciclazocina/análogos & derivados , Receptores Opioides/metabolismo , Animales , Células CHO , Cricetinae , Cricetulus , Ciclazocina/química , Ciclazocina/metabolismo , Humanos , Unión Proteica/fisiología , Relación Estructura-ActividadRESUMEN
Improgan, a chemical congener of cimetidine, is a highly effective non-opioid analgesic when injected into the CNS. Despite extensive characterization, neither the improgan receptor, nor a pharmacological antagonist of improgan has been previously described. Presently, the specific binding of [(3)H]cimetidine (3HCIM) in brain fractions was used to discover 4(5)-((4-iodobenzyl)thiomethyl)-1H-imidazole, which behaved in vivo as the first improgan antagonist. The synthesis and pharmacological properties of this drug (named CC12) are described herein. In rats, CC12 (50-500nmol, i.c.v.) produced dose-dependent inhibition of improgan (200-400nmol) antinociception on the tail flick and hot plate tests. When given alone to rats, CC12 had no effects on nociceptive latencies, or on other observable behavioral or motor functions. Maximal inhibitory effects of CC12 (500nmol) were fully surmounted with a large i.c.v. dose of improgan (800nmol), demonstrating competitive antagonism. In mice, CC12 (200-400nmol, i.c.v.) behaved as a partial agonist, producing incomplete improgan antagonism, but also limited antinociception when given alone. Radioligand binding, receptor autoradiography, and electrophysiology experiments showed that CC12's antagonist properties are not explained by activity at 25 sites relevant to analgesia, including known receptors for cannabinoids, opioids or histamine. The use of CC12 as an improgan antagonist will facilitate the characterization of improgan analgesia. Furthermore, because CC12 was also found presently to inhibit opioid and cannabinoid antinociception, it is suggested that this drug modifies a biochemical mechanism shared by several classes of analgesics. Elucidation of this mechanism will enhance understanding of the biochemistry of pain relief.
Asunto(s)
Cimetidina/análogos & derivados , Antagonistas de los Receptores H2 de la Histamina/metabolismo , Imidazoles/farmacología , Receptores Histamínicos H2/efectos de los fármacos , Sulfuros/farmacología , Analgésicos Opioides/farmacología , Animales , Autorradiografía , Benzoxazinas/farmacología , Sitios de Unión/efectos de los fármacos , Cimetidina/antagonistas & inhibidores , Cimetidina/metabolismo , Cimetidina/farmacología , Relación Dosis-Respuesta a Droga , Encefalina Ala(2)-MeFe(4)-Gli(5)/farmacología , Guanosina 5'-O-(3-Tiotrifosfato)/farmacología , Histamina/farmacología , Imidazoles/síntesis química , Indicadores y Reactivos , Inyecciones Intraventriculares , Ligandos , Masculino , Membranas/efectos de los fármacos , Membranas/metabolismo , Ratones , Morfolinas/farmacología , Naloxona/farmacología , Naftalenos/farmacología , Antagonistas de Narcóticos/farmacología , Dimensión del Dolor/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Sulfuros/síntesis químicaRESUMEN
The synthesis and evaluation of a series of aryl-containing N-monosubstituted analogues of the lead compound 8-carboxamidocyclazocine were performed to probe a putative hydrophobic binding pocket of opioid receptors. High binding affinity to mu, kappa, and delta opioid receptors was observed for the 8-[N-(4'-phenyl)-phenethyl)carboxamido] analogue.
Asunto(s)
Ciclazocina/análogos & derivados , Ciclazocina/síntesis química , Receptores Opioides delta/efectos de los fármacos , Receptores Opioides kappa/efectos de los fármacos , Receptores Opioides mu/efectos de los fármacos , Animales , Células CHO , Cricetinae , Cricetulus , Ciclazocina/farmacología , Etilcetociclazocina/análogos & derivados , Etilcetociclazocina/síntesis química , Etilcetociclazocina/farmacología , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Ensayo de Unión Radioligante , Receptores Opioides delta/metabolismo , Receptores Opioides kappa/metabolismo , Receptores Opioides mu/metabolismo , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
The antinociceptive profile of selected histamine H(2) and histamine H(3) receptor antagonists led to the discovery of improgan, a non-brain-penetrating analgesic agent which does not act on known histamine receptors. Because no chemical congener of improgan has yet been discovered which has both antinociceptive and brain-penetrating properties, the present study investigated the antinociceptive effects of a series of chemical compounds related to zolantidine, a brain-penetrating histamine H(2) receptor antagonist. The drugs studied presently contain the piperidinomethylphenoxy (PMPO) moiety, hypothesized to introduce brain-penetrating characteristics. Following intracerebroventricular (i.c.v.) dosing in rats, six of eight drugs produced dose- and time-related antinociception on both the tail flick and hot plate tests over a nearly eight-fold range of potencies. Ataxia and other motor side effects were observed after high doses of these drugs, but two of the compounds (SKF94674 and loxtidine) produced maximal antinociception at doses which were completely devoid of these motor effects. Consistent with the hypothesis that PMPO-containing drugs are brain-penetrating analgesics, SKF94674 and another derivative (JB-9322) showed dose-dependent antinociceptive activity 15 to 30 min after systemic dosing in mice, but these effects were accompanied by seizures and death beginning 45 min after dosing. Other drugs showed a similar pattern of antinociceptive and toxic effects. In addition, loxtidine produced seizures without antinociception, whereas zolantidine produced neither effect after systemic dosing in mice. Although several of the drugs tested have histamine H(2) receptor antagonist activity, neither the antinociception nor the toxicity was correlated with histamine H(2) receptor activity. The present results are the first to demonstrate the existence of brain-penetrating antinociceptive agents chemically related to zolantidine and improgan, but further studies are needed to understand the mechanisms of both the pain relief and toxicity produced by these agents.
Asunto(s)
Analgésicos/farmacología , Encéfalo/metabolismo , Cimetidina/análogos & derivados , Algoritmos , Analgésicos/química , Analgésicos/farmacocinética , Animales , Conducta Animal/efectos de los fármacos , Benzotiazoles , Cimetidina/química , Cimetidina/farmacología , Relación Dosis-Respuesta a Droga , Antagonistas de los Receptores H2 de la Histamina/química , Antagonistas de los Receptores H2 de la Histamina/farmacología , Inyecciones Intraventriculares , Masculino , Ratones , Estructura Molecular , Dolor/prevención & control , Dimensión del Dolor/métodos , Fenoxipropanolaminas/química , Fenoxipropanolaminas/farmacología , Piperidinas/química , Piperidinas/farmacología , Quinazolinas/química , Quinazolinas/farmacología , Quinazolinonas , Ratas , Ratas Sprague-Dawley , Tiazoles/química , Tiazoles/farmacología , Factores de Tiempo , Triazoles/química , Triazoles/farmacologíaRESUMEN
8-Position variants of cyclazocine have been made where the phenolic 8-OH was replaced by thioamide, amidine, guanidine, urea and thiourea groups. High affinity for opioid receptors was observed for the 8-CSNH2 and 8-NHCHS analogues indicating that these groups are isosteric with not only the 8-OH but with the previously synthesized 8-CONH2 and 8-NHCHO cyclazocine derivatives.
Asunto(s)
Ciclazocina/química , Ciclazocina/farmacología , Relación Estructura-ActividadRESUMEN
Very high affinity for opioid receptors (e.g., K(i)=0.052nM for mu) has been observed in the rationally designed naltrexone analogue 5. SAR and physical data supports the hypothesis that the 4-OH group of 5 stabilizes the 3-carboxamido group in the putative bioactive conformation.
Asunto(s)
Naltrexona/análogos & derivados , Naltrexona/metabolismo , Receptores Opioides/metabolismo , Animales , Células CHO , Cricetinae , Humanos , Naltrexona/síntesis química , Unión Proteica/fisiologíaRESUMEN
The present study evaluated the effects of 8-carboxamidocyclazocine (8-CAC), a novel mixed-action kappa/mu agonist with a long duration of action, on food- and cocaine-maintained responding in rhesus monkeys to assess the potential utility of 8-CAC as a medication for the treatment of cocaine dependence. The effects of acute and chronic (10 days) 8-CAC were examined in rhesus monkeys responding under a multiple schedule for both cocaine and food reinforcement. Acute 8-CAC (0.032-0.56 mg/kg, i.m.) dose-dependently eliminated cocaine-maintained responding in all three monkeys. However, doses of 8-CAC that decreased cocaine self-administration typically also decreased food-maintained responding, and 8-CAC-induced decreases in cocaine self-administration diminished during chronic 8-CAC treatment. These results confirm that 8-CAC acutely decreases cocaine self-administration. However, non-selective effects of 8-CAC on food-maintained responding and tolerance to 8-CAC effects on cocaine self-administration may limit its potential for the treatment of cocaine dependence.
Asunto(s)
Cocaína/farmacología , Condicionamiento Operante/efectos de los fármacos , Ciclazocina/análogos & derivados , Ciclazocina/farmacología , Alimentos , Receptores Opioides kappa/agonistas , Receptores Opioides mu/agonistas , Recompensa , Animales , Relación Dosis-Respuesta a Droga , Femenino , Hipnóticos y Sedantes/farmacología , Macaca mulatta , Masculino , Esquema de Refuerzo , Vómitos/inducido químicamenteRESUMEN
Cocaine abuse has been implicated as a cofactor in human immunodeficiency virus (HIV)-1-associated dementia (HAD). In this study, we tested the hypothesis that exposure of microglial cells, the resident macrophages of the brain, to cocaine would potentiate HIV-1 expression. Because kappa-opioid receptor (KOR) agonists have been shown to suppress neurochemical and neurobehavioral responses to cocaine and to inhibit HIV-1 expression in microglial cell cultures, we also postulated that KOR ligands would inhibit cocaine-induced potentiation of HIV-1 expression. Human microglial cells were infected with HIV-1(SF162), an R5 isolate, and viral expression was quantified by measurement of p24 antigen in culture supernatants. Treatment of microglia with the KOR agonists trans-(+/-)-3,4-dichlor-N-methyl-N-(2[1-pyrrolidnyl])benzeneacetamide methanesulfonate and 8-carboxamidocyclazocine inhibited viral expression (maximal suppression of 42 and 48%, respectively). Consistent with the hypotheses, treatment of microglia with cocaine promoted HIV-1 expression (maximal enhancement of 54%), and pretreatment of microglia with these KOR agonists as well as with the KOR-selective antagonist nor-binaltorphimine abrogated cocaine-induced potentiation of viral expression. Results of flow cytometry studies suggested that the mechanism whereby KOR ligands inhibit cocaine's stimulatory effect on viral expression involves the suppression of cocaine-induced activation of extracellular signal-regulated kinase1/2, thereby blunting cocaine-enhanced up-regulation of the HIV-1 entry chemokine coreceptor CCR5. The findings of this study suggest that in addition to its neurotoxic effects, cocaine could foster development of HAD by potentiating viral expression in the brain and that this phenomenon is inhibited by KOR ligands.
Asunto(s)
Analgésicos Opioides/farmacología , Cocaína/farmacología , VIH-1/efectos de los fármacos , Microglía/efectos de los fármacos , Receptores Opioides kappa/agonistas , 3,4-Dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclohexil)-bencenacetamida, (trans)-Isómero/farmacología , Células Cultivadas , Activación Enzimática/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , VIH-1/fisiología , Humanos , Microglía/virología , Proteínas Quinasas Activadas por Mitógenos/antagonistas & inhibidores , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Receptores CCR5/metabolismo , Receptores Opioides kappa/metabolismo , Regulación hacia Arriba/efectos de los fármacos , Replicación Viral/efectos de los fármacosRESUMEN
A series of 10-ketomorphinan analogues were synthesized, and their binding affinity at all three opioid receptors was investigated. In most cases, high affinity at micro and kappa receptors, and lower affinity at delta receptor was observed, resulting in good selectivity for micro and kappa receptors. A wide range of substituents can be accommodated on the nitrogen position. The N-(S)-tetrahydrofurfuryl analogue 11 displayed the highest affinity at all three receptors. The N-cyclobutylmethyl analogue 13 gave both high affinity and selectivity at kappa receptor, and N-2-phenylethyl analogue 18 exhibited good affinity and selectivity at micro receptor. Further modifications of the 3-substituent indicated that one H-bond donor was an essential requirement for good affinity at micro and kappa receptors. Similar modifications were investigated at the 3-OH group of morphinans: levorphanol (2a), cyclorphan (2b), and MCL-101 (2c) lacking the 10-keto group. The 3-amino bioisosteric analogues (40 and 41) displayed reasonably good affinity at micro and kappa receptors. The 3-carboxamido replacement (compounds 46-48) in the morphinan subseries resulted in similar affinities comparable to their corresponding 3-OH congeners. The high affinity of these carboxamido analogues, along with their greater lipophilicity and metabolic stability, make them promising candidates for further pharmacological investigation.