RESUMO
Spinocerebellar ataxias (SCAs) are a group of genetic diseases characterized by progressive ataxia and neurodegeneration, often in cerebellar Purkinje neurons. A SCA1 mouse model, Pcp2-ATXN1[30Q]D776, has severe ataxia in absence of progressive Purkinje neuron degeneration and death. Previous RNA-seq analyses identify cerebellar upregulation of the peptide hormone cholecystokinin (Cck) in Pcp2-ATXN1[30Q]D776 mice. Importantly, absence of Cck1 receptor (Cck1R) in Pcp2-ATXN1[30Q]D776 mice confers a progressive disease with Purkinje neuron death. Administration of a Cck1R agonist, A71623, to Pcp2-ATXN1[30Q]D776;Cck-/- and Pcp2-AXTN1[82Q] mice dampens Purkinje neuron pathology and associated deficits in motor performance. In addition, A71623 administration improves motor performance of Pcp2-ATXN2[127Q] SCA2 mice. Moreover, the Cck1R agonist A71623 corrects mTORC1 signaling and improves expression of calbindin in cerebella of AXTN1[82Q] and ATXN2[127Q] mice. These results indicate that manipulation of the Cck-Cck1R pathway is a potential therapeutic target for treatment of diseases involving Purkinje neuron degeneration.
Assuntos
Quimiocinas CC/agonistas , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Células de Purkinje/efeitos dos fármacos , Ataxias Espinocerebelares/tratamento farmacológico , Tetragastrina/análogos & derivados , Animais , Ataxina-1/genética , Ataxina-1/metabolismo , Atrofia , Comportamento Animal/efeitos dos fármacos , Calbindinas/metabolismo , Quimiocinas CC/genética , Quimiocinas CC/metabolismo , Colecistocinina/genética , Colecistocinina/metabolismo , Modelos Animais de Doenças , Feminino , Predisposição Genética para Doença , Fatores de Troca do Nucleotídeo Guanina/genética , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Atividade Motora/efeitos dos fármacos , Degeneração Neural , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Células de Purkinje/enzimologia , Células de Purkinje/patologia , Transdução de Sinais , Ataxias Espinocerebelares/enzimologia , Ataxias Espinocerebelares/genética , Ataxias Espinocerebelares/patologia , Tetragastrina/farmacologiaRESUMO
Pain is associated with negative emotions such as anxiety, but the underlying neurocircuitry and modulators of the association of pain and anxiety remain unclear. The neuropeptide cholecystokinin (CCK) has both pronociceptive and anxiogenic properties, so we explored the role of CCK in anxiety and nociception in the central amygdala (CeA), a key area in control of emotions and descending pain pathways. Local infusion of CCK into the CeA of control rats increased anxiety, as measured in the light-dark box test, but had no effect on mechanical sensitivity. By contrast, intra-CeA CCK infusion 4 days after Complete Freund's Adjuvant (CFA) injection into the hindpaw resulted in analgesia, but also in loss of its anxiogenic capacity. Inflammatory conditions induced changes in the CeA CCK signaling system with an increase of CCK immunoreactivity and a decrease in CCK1, but not CCK2, receptor mRNA. In CFA rats, patch-clamp experiments revealed that CCK infusion increased CeA neuron excitability. It also partially blocked the discharge of wide dynamic range neurons in the dorsal spinal cord. These effects of CCK on CeA and spinal neurons in CFA rats were mimicked by the specific CCK2 receptor agonist, gastrin. This analgesic effect was likely mediated by identified CeA neurons projecting to the periaqueductal gray matter that express CCK receptors. Together, our data demonstrate that intra-CeA CCK infusion activated a descending CCK2 receptor-dependent pathway that inhibited spinal neuron discharge. Thus, persistent pain induces a functional switch to a newly identified analgesic capacity of CCK in the amygdala, indicating central emotion-related circuit controls pain transmission in spinal cord.
Assuntos
Tonsila do Cerebelo/metabolismo , Colecistocinina/metabolismo , Dor/patologia , Receptor de Colecistocinina B/metabolismo , Transdução de Sinais/fisiologia , Tonsila do Cerebelo/patologia , Animais , Adaptação à Escuridão/efeitos dos fármacos , Modelos Animais de Doenças , Comportamento Exploratório/efeitos dos fármacos , Adjuvante de Freund/toxicidade , Gastrinas/uso terapêutico , Glutamato Descarboxilase/metabolismo , Inflamação/induzido quimicamente , Inflamação/complicações , Masculino , Neurônios/efeitos dos fármacos , Neurônios/fisiologia , Nociceptividade/efeitos dos fármacos , Dor/etiologia , Limiar da Dor/efeitos dos fármacos , Substância Cinzenta Periaquedutal/efeitos dos fármacos , Substância Cinzenta Periaquedutal/fisiologia , Ratos , Ratos Sprague-Dawley , Receptor de Colecistocinina B/agonistas , Receptor de Colecistocinina B/antagonistas & inibidores , Receptor de Colecistocinina B/genética , Transdução de Sinais/efeitos dos fármacos , Sincalida/uso terapêutico , Tetragastrina/análogos & derivados , Tetragastrina/uso terapêuticoRESUMO
INTRODUCCIÓN: El presente dictamen expone la evaluación de la eficacia y seguridad de carfilzomib en combinación con lenalidomida y dexametasona para pacientes adultos con mieloma múltiple en recaída y/o refractario, inelegible o posterior a trasplante autólogo de progenitores hematopoyéticos, que han recibido al menos dos líneas de tratamiento previo. El mieloma múltiple (MM) es una proliferación neoplásica de un clon único de células plasmáticas de causa desconocida. En Perú, se habrían presentado 995 casos de MM en 2018, representando el 1.7 % del total de casos de cáncer para ese año. Durante el curso de la enfermedad, es esperable que el paciente con MM presente recaída o refractariedad a la terapia específica para la enfermedad, existiendo alternativas terapéuticas que incluyen al trasplante de células hematopoyéticas, uso de nuevos regímenes de tratamiento a los que el paciente no haya estado expuesto previamente, repetir régimen quimioterapéutico utilizado anteriormente, o una terapia experimental ofrecida como parte de un ensayo clínico. En EsSalud, se dispone de lenalidomida 5 mg para el tratamiento del MM en pacientes que hayan recibido al menos un tratamiento previo y para pacientes postrasplante autólogo de progenitores hematopoyéticos, lenalidomida 25 mg para pacientes con MM que fracasaron a primera y segunda línea de tratamiento y para pacientes postrasplante autólogo de progenitores hematopoyéticos; también se dispone de talidomida 100 mg con indicación de uso en hematología y oncología médica y bortezomib 3.5 mg para el paciente con MM con compromiso renal y para el paciente con MM recidivante y/o refractario a uno o más tratamientos previos. Si bien se dispone de estos productos farmacéuticos, existe interés por terapias para el MM en recaída y/o refractario, inelegible o posterior a trasplante autólogo, que han recibido al menos dos líneas de tratamiento, por lo que algunos médicos especialistas solicitan evaluación de uso de carfilzomib, producto farmacológico no incluido en el Petitorio Farmacológico de EsSalud, asociado a lenalidomida y dexametasona, para la mejora en la sobrevida global (SG) de estos pacientes. OBJETIVO: El objetivo del presente dictamen fue la evaluación de la mejor evidencia disponible sobre la eficacia y seguridad de carfilzomib en combinación con lenalidomida y dexametasona comparada con lenalidomida y dexametasona para pacientes adultos con MM en recaída y/o refractario, inelegible o posterior a trasplante autólogo, que han recibido al menos dos líneas de tratamiento previo (bortezomib y lenalidomida o talidomida). TECNOLOGÍA SANITARIA DE INTERÉS: Carfilzomib (KYPROLIS®), producido por la compañía Amgen Inc., es un tetrapéptido con un grupo epoxicetona inhibidor del proteosoma que se une de forma selectiva e irreversible a la treonina en el extremo N terminal de los sitios activos del proteosoma 20S (núcleo proteolítico del proteosoma 26S) y que muestra poca o ninguna actividad frente a otros tipos de proteasas. La inhibición generada por carfilzomib produce actividad antiproliferativa y proapoptótica en modelos preclínicos de tumores hematológicos. METODOLOGÍA: Se realizó una búsqueda sistemática de literatura con el objetivo de identificar la mejor evidencia disponible sobre la eficacia y seguridad de carfilzomib en combinación con lenalidomida y dexametasona comparada con lenalidomida y dexametasona para pacientes con MM en recaída y/o refractario, inelegible o posterior a trasplante autólogo de progenitores hematopoyéticos, que han recibido al menos dos líneas de tratamiento previo. RESULTADOS: Las GPC de NCCN, IMWG y Mayo Clinic recomiendan el empleo del esquema triple de carfilzomib más lenalidomida más dexametasona en pacientes con MM en recaída, que han recibido al menos dos líneas de tratamiento previo. Específicamente, IMWG y Mayo Clinic lo recomiendan en los casos de recaída y refractariedad a lenalidomida y dexametasona (doble refractariedad), mientras que NCCN no brinda detalles de la recomendación. Adicionalmente, NCCN y IMWG recomiendan el esquema terapéutico de lenalidomida más dexametasona (disponible en EsSalud) en pacientes ancianos o frágiles, o en casos de enfermedad indolente, respectivamente. Además, recomiendan repetir un régimen (e.g. lenalidomida más dexametasona) si este fue utilizado como terapia de inducción y la recaída se dio posterior a los seis meses. Por su parte, ESMO recomienda otros esquemas de tratamiento que no forman parte de la presente evaluación. CONCLUSIONES: En el presente documento, se evaluó la mejor evidencia científica disponible hasta la actualidad sobre la eficacia y seguridad de carfilzomib en combinación con lenalidomida y dexametasona para pacientes con MM en recaída y/o refractario, inelegible o posterior a trasplante autólogo, que han recibido al menos dos líneas de tratamiento previo (bortezomib y lenalidomida o talidomida). ï La principal evidencia que responde a nuestra pregunta PICO procede del estudio ASPIRE, ensayo clínico aleatorizado, de etiqueta abierta, controlado de fase III, que buscó evaluar los beneficios adicionales en seguridad y eficacia de la terapia con carfilzomib asociado a lenalidomida y dexametasona comparado con lenalidomida más dexametasona en pacientes adultos con MM en recaída que habían recibido de una a tres líneas de terapia previa. En los hallazgos del análisis final de SG, con una mediana de seguimiento de sobrevida global de 67.1 meses, no se observó una diferencia estadísticamente significativa en el riesgo de morir entre ambos grupos (grupo de carfilzomib: 246/396 muertes [62.1 %], grupo control: 267/396 muertes [67.4 %], RR = 0.92, IC 95 %: 0.83-1.02, p = 0.118, calculado por el equipo técnico IETSI). Sin embargo, es importante mencionar que el uso diferenciado de terapias subsecuentes pudo hacer influenciado sobre los resultados de SG o mortalidad. Por otro lado, no se encontró una diferencia clínicamente relevante en la calidad de vida entre ambos grupos de estudio (diferencia menor a 5 puntos) en el ciclo 18 de tratamiento. De esta forma, existe incertidumbre sobre el beneficio ganado con el uso del esquema tiple de carfilzomib en desenlaces clínicamente relevantes desde la perspectiva del paciente como SG y calidad de vida. En cuanto a la seguridad, los pacientes en el grupo carfilzomib presentaron un mayor número de eventos adversos serios. Además, existió una gran incertidumbre en relación a la seguridad a largo plazo con carfilzomib ya que su uso fue evaluado hasta los 18 meses de tratamiento, mientras que según la etiqueta de EMA este régimen podría ser utilizado por plazos mayores (hasta la progresión). Respecto a la aplicabilidad de los resultados del estudio ASPIRE, se debe mencionar que la población del estudio ASPIRE no fue representativa de la población de la pregunta PICO (ya que la gran mayoría de pacientes no habían recibido previamente bortezomib y lenalidomida o talidomida), lo que limita la extrapolación de los resultados para el grupo de pacientes de interés. Por otro lado, el empleo de un diseño de etiqueta abierta y desarrollo del estudio por parte de la compañía que comercializa el producto son posibles fuentes de sesgos que podrían influir en la precisión de los resultados reportados por los autores. En conclusión, no se ha podido determinar un beneficio del esquema triple de carfilzomib en cuanto a la SG, ni se ha encontrado que este sea superior en términos de calidad de vida, ni que presente un mejor perfil de seguridad en comparación con lenalidomida y dexametasona. Estos hallazgos impiden que se pueda sustentar técnicamente una recomendación favorable para el uso de este medicamento. El IETSI no aprueba el uso de carfilzomib asociado a lenalidomida y dexametasona para pacientes adultos con mieloma múltiple en recaída y/o refractario, inelegible o posterior a trasplante autólogo de progenitores hematopoyéticos, que han recibido al menos dos líneas de tratamiento previo.
Assuntos
Humanos , Tetragastrina/análogos & derivados , Dexametasona/uso terapêutico , Lenalidomida/uso terapêutico , Mieloma Múltiplo/tratamento farmacológico , Análise Custo-Eficiência , Combinação de Medicamentos , Avaliação de MedicamentosRESUMO
The conformational analysis with 1H NMR spectroscopy method in solution and the structure-activity relationship study of a series sterically restricted analogs allowed to detect the possible biologically active conformation of N-(6-phenylhexanoyl)glycyl-tryptophan amide (GB-115), a highly active dipeptide cholecystokinin-4 analog with anxiolytic activity. The structure-activity relationship study of GB-115 and the series of its' glycine- and proline-containing analogs with different C-terminal substitute detected the anxiolytic activity of compounds with beta-turn like conformation and inactivity of compounds with gamma-turn like conformation. So, the GB-115 biologically active conformation is beta-turn. The results of nuclear Overhauser effect study permitted to qualify the betaII-turn conformation as GB-115 biologically active conformation. The following synthesis of sterically restricted GB-115 analogs (2S)-2-{(3R)-3-[(6-phenylhexanoyl)amino]-2-oxopyrrolidin-1-yl}-3-(1H-indol-3-yl)propionic acid ethyl ester, N-(6-phenylhexanoyl)glycyl-N(alpha)(methyl)-tryptophan ethyl ester, (2S)-2-[10,11-dihydro-5H-dibenzo[b,f] azepin-5-carbonyl)-amino]-3-(1H-indol-3-yl)propionic acid methyl ester and (2S)-2-[({3-[(ethoxycarbonyl)amino]-10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl}carbonyl)amino]-3-(1H-indol-3-yl)propionic acid methyl ester confirmed the estimated type of GB-115 biologically active conformation.
Assuntos
Ansiedade/tratamento farmacológico , Dipeptídeos/química , Relação Estrutura-Atividade , Tetragastrina/química , Animais , Ansiolíticos/química , Ansiolíticos/farmacologia , Ansiedade/patologia , Dipeptídeos/síntese química , Dipeptídeos/farmacologia , Glicina/química , Humanos , Espectroscopia de Ressonância Magnética , Prolina/química , Estrutura Secundária de Proteína , Ratos , Tetragastrina/análogos & derivados , Tetragastrina/farmacologia , Triptofano/químicaRESUMO
To test whether a turnlike arrangement is involved in the bioactive conformation of CCK4 analogues upon CCK1 receptor recognition, we describe the preparation of two series of CCK4 derivatives, in which the central dipeptide Met-Asp has been replaced by recognized beta-turn mimetics {(2S,5S,11bR)- and (2R,5R,11bS)-2-amino-5-carboxy-3-oxo-2,3,5,6,11,11b-hexahydro-1H-indolizino[8,7-b]indole (IBTM) and beta-turn dipeptide, 2-oxo-7-thio-1-azabicyclo[4.3.0]nonane (BTD)}. The incorporation of the indolizinoindole IBTM type II beta-turn mimetic is preferred over its type II' counterpart for efficient CCK1 receptor recognition, while BTD derivatives were completely inactive. The structure-conformation-activity relationship study in the IBTM series has shown some essential requirement of these CCK4 derivatives to favorably interact with CCK1 receptors: (a) the adoption of turnlike conformations, (b) the presence of an L-Phe residue and a C-terminal carboxamide moiety, and (c) the indole ring of the IBTM skeleton. Moreover, the existence of pi-pi interactions between the phenyl ring of d-Phe residues and the indole ring of IBTM framework is detrimental for binding affinity. A series of potent and selective CCK1 receptor antagonists, exemplified by compounds 8a and 8b, emerges among these IBTM-containing derivatives.
Assuntos
Compostos Bicíclicos Heterocíclicos com Pontes/química , Dipeptídeos/química , Indóis/química , Tetragastrina/análogos & derivados , Tetragastrina/síntese química , Amilases/metabolismo , Animais , Córtex Cerebral/metabolismo , Técnicas In Vitro , Ligantes , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Mimetismo Molecular , Pâncreas/efeitos dos fármacos , Pâncreas/metabolismo , Estrutura Secundária de Proteína , Ensaio Radioligante , Ratos , Receptor de Colecistocinina A/antagonistas & inibidores , Relação Estrutura-Atividade , Tetragastrina/química , Tetragastrina/farmacologiaRESUMO
Analogues of the endogenous peptide corresponding to the 30-33 sequence of cholecystokinin (Trp-Met-Asp-Phe-NH2) were synthesized, and their biological activity was studied. It was shown that, in rats, the N-succinylated Nle2 analogue of this tetrapeptide exhibits increased anxiolytic properties in the dark-bright chamber test and an enhanced alcohol intake by both the control animals and the long-time alcohol-dependent animals under the conditions of free choice. Introduction of an isopropyl residue into the C-terminal amide of the Nle2 analogue resulted in the appearance of anxiolytic and antialcohol activity and the ability to increase the morphine analgesic effect in the tail-flick test on rats. The two synthesized analogues retained an affinity to cholecystokinin receptors.
Assuntos
Consumo de Bebidas Alcoólicas/tratamento farmacológico , Comportamento Animal/efeitos dos fármacos , Oligopeptídeos/síntese química , Tetragastrina/análogos & derivados , Tetragastrina/síntese química , Analgésicos/síntese química , Analgésicos/farmacologia , Animais , Ansiolíticos/síntese química , Ansiolíticos/farmacologia , Ansiedade/induzido quimicamente , Encéfalo/metabolismo , Sinergismo Farmacológico , Técnicas In Vitro , Morfina/farmacologia , Oligopeptídeos/farmacologia , Pâncreas/metabolismo , Ensaio Radioligante , Ratos , Ratos Wistar , Receptor de Colecistocinina A/metabolismo , Receptor de Colecistocinina B/metabolismo , Relação Estrutura-Atividade , Tetragastrina/farmacologia , Triptofano/químicaRESUMO
The influence of intraventricular cholecystokinin-8S (CCK-8S) and systemic N-t-Boc-Trp-Met-Asp-Phe-amide (Boc CCK-4) was evaluated in the acoustic and fear-potentiated startle paradigms in CD-1 mice. In the light + tone startle condition. CCK-8S increased startle 168 hr after administration, compared with saline. In the tone startle condition, CCK-8S decreased startle immediately and 24 hr after administration, compared with saline. Among nonshocked mice, CCK-8S increased startle at 48 and 168 hr, compared with saline. In the light + tone condition, 5 microg Boc-CCK-4 did not influence startle, whereas 15 microg Boc CCK-4 decreased startle immediately, 24 hr, and 48 hr following administration. Results demonstrate that antecedent environmental experiences interact with subsequent pharmacological challenges in provoking the temporal expression of alterations in startle magnitude.
Assuntos
Nootrópicos/farmacologia , Reflexo de Sobressalto/efeitos dos fármacos , Sincalida/análogos & derivados , Sincalida/farmacologia , Tetragastrina/análogos & derivados , Tetragastrina/farmacologia , Estimulação Acústica , Animais , Medo , Infusões Intravenosas , Injeções Intraventriculares , Masculino , Camundongos , Nootrópicos/administração & dosagem , Sincalida/administração & dosagem , Tetragastrina/administração & dosagemRESUMO
2-Oxopiperazine derivatives 1 have been designed as mimetics of gamma-turn conformationally constrained tripeptides. The synthetic pathway devised for the preparation of both epimers of 1 at C(5) involves a reductive amination of cyanomethyleneamino pseudopeptides with amino acid derivatives, followed by regiospecific lactamization of the resulting C-backbone branched pseudopeptides. The versatility of this methodology is illustrated in the synthesis of analogues of the tetrapeptides Boc-[Nle(31)]-CCK-4 and Boc-[Lys(o-tolylaminocarbonyl)(31)]-CCK-4. The introduction of the new conformational restriction into these Boc-CCK-4 analogues led to a loss of 2 or 3 orders of magnitude in the affinity at CCK receptors. These results suggest the absence of a gamma-turn in the bioactive conformation of the C-terminal tripeptide of CCK-4.
Assuntos
Peptídeos/síntese química , Piperazinas/química , Tetragastrina/análogos & derivados , Animais , Córtex Cerebral/metabolismo , Modelos Moleculares , Mimetismo Molecular , Ressonância Magnética Nuclear Biomolecular , Pâncreas/metabolismo , Peptídeos/química , Peptídeos/metabolismo , Piperazinas/síntese química , Piperazinas/metabolismo , Conformação Proteica , Ratos , Receptor de Colecistocinina B , Receptores da Colecistocinina/metabolismo , Tetragastrina/metabolismoRESUMO
Latent inhibition (LI) is a behavioural paradigm in which repeated exposure to a stimulus without consequence inhibits the formation of any new associations with that stimulus. To the extent that LI reflects a process of leaming to ignore irrelevant stimuli, disrupted LI has been suggested as an animal model for the attentional deficits observed in schizophrenia. The antipsychotic potential of cholecystokinin (CCK) stems from its colocalization with dopamine (DA) in the mesolimbic pathway, where it demonstrates both excitatory and inhibitory effects on dopaminergic activity. This may be explained by mediation through different receptor subtypes. A variety of hypotheses has emerged regarding the potential clinical application of subtype-selective CCK-based drugs. The present experiments examined the effects on LI of two selective CCK(A) ligands: PD-140,548 (a CCK(A) antagonist, Experiment 1: 0.001, 0.01, and 0.1 mg/kg) and A-71623 (a CCK(A) agonist, Experiment 2: 0.02, 0.05, and 0.1 mg/kg). In both experiments, the effects of haloperidol (0.1 mg/kg) were also investigated. Animals receiving 0.1 mg/kg of haloperidol or 0.001 or 0.1 mg/kg (but not 0.01 mg/kg) of PD-140,548 treated the preexposed stimulus as irrelevant after a low number of preexposures. In contrast, no facilitatory effect on LI was detectable at any of the A-71623 doses. The finding that A-71623 failed to enhance LI indicates that it is unlikely that this compound would have any antipsychotic effect within the clinical setting. Considering the facilitatory effect exerted by PD-140,548 on LI, it is probable that the inhibition of CCK activity might prove a more promising strategy for the pharmacological treatment of schizophrenia.
Assuntos
Indóis/farmacologia , Inibição Psicológica , Receptores da Colecistocinina/agonistas , Receptores da Colecistocinina/antagonistas & inibidores , Tetragastrina/análogos & derivados , Tetragastrina/farmacologia , Animais , Haloperidol/farmacologia , Indóis/metabolismo , Ligantes , Masculino , Ratos , Ratos Sprague-Dawley , Receptor de Colecistocinina A , Receptores da Colecistocinina/metabolismo , Receptores da Colecistocinina/fisiologia , Tetragastrina/metabolismoRESUMO
CCK released by intraluminal stimuli modifies duodenal activity contributing to a decrease in gastric emptying. However, the neural mechanisms by which CCK controls motility are not well known. The aim of this study was to investigate the interaction between CCK and the enteric nervous system through the study of the effects of CCK-8 on ascending excitation. Anaesthetized Sprague-Dawley rats were prepared with a strain-gauge sutured to the duodenum wall. An electrode holder was placed in the duodenum lumen to elicit ascending contraction. Electrical field stimulation of the duodenal mucosa (4 Hz, 0.6 ms, 30 V) induced an ascending excitation which was blocked by hexamethonium (10 mg kg-1; n=5) and atropine (0.3 mg kg-1; n=5), but enlarged by L-NNA (10(-5) mol kg-1; n=5). CCK-8 (3 ¿ 10(-9) mol kg-1 10 min-1) blocked ascending excitation and an inhibition of the induced phasic activity was observed instead (n=18). Individually, none of the CCK receptor antagonists (L-364 718 and L-365 260) (3 ¿ 10(-7) mol kg-1; n=6 each) blocked the inhibition of ascending excitation induced by CCK-8. However, simultaneous infusion of both antagonists abolished CCK-8 effect on electrical stimulation (n=5). Similarly, none of the CCK-8 agonists (A-71623, A-71378, gastrin) modified the ascending excitation. In contrast, the simultaneous infusion of A-71623 and CCK-4 (n=4) induced an effect similar to CCK-8. In conclusion, CCK-8 blocked ascending contraction elicited by electrical field stimulation of duodenal mucosa by means of simultaneous activation of CCK-A and CCK-B receptors.
Assuntos
Colecistocinina/farmacologia , Duodeno/efeitos dos fármacos , Sistema Nervoso Entérico/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Contração Muscular/efeitos dos fármacos , Peristaltismo/efeitos dos fármacos , Antagonistas Adrenérgicos beta/farmacologia , Animais , Atropina/farmacologia , Benzodiazepinonas/farmacologia , Colecistocinina/agonistas , Colecistocinina/antagonistas & inibidores , Depressão Química , Devazepida/farmacologia , Duodeno/fisiologia , Estimulação Elétrica , Sistema Nervoso Entérico/fisiologia , Inibidores Enzimáticos/farmacologia , Gastrinas/farmacologia , Hexametônio/farmacologia , Mucosa Intestinal/fisiologia , Masculino , Antagonistas Nicotínicos/farmacologia , Nitroarginina/farmacologia , Oligopeptídeos/farmacologia , Parassimpatolíticos/farmacologia , Fentolamina/farmacologia , Compostos de Fenilureia/farmacologia , Propranolol/farmacologia , Ratos , Ratos Sprague-Dawley , Receptor de Colecistocinina A , Receptor de Colecistocinina B , Receptores da Colecistocinina/efeitos dos fármacos , Receptores da Colecistocinina/fisiologia , Receptores de Peptídeo Intestinal Vasoativo/antagonistas & inibidores , Sincalida/farmacologia , Tetragastrina/análogos & derivados , Tetragastrina/farmacologia , Peptídeo Intestinal Vasoativo/análogos & derivados , Peptídeo Intestinal Vasoativo/farmacologiaRESUMO
Cholecystokinin (CCK) regulates intestinal motility after being released by several luminal nutrients. However the mechanism of action of CCK is still not well known. The aim of our study was to establish the mechanism of action of CCK in the rat intestine using an in vivo model and focusing on the nervous pathways involved in the response as well as type of receptors. Anesthetized rats were prepared with two strain-gauges, in duodenum and jejunum, to record circular muscle motor activity. A group of animals was also prepared with a catheter to infuse capsaicin inside the duodenum. Responses to CCK-octapeptide (CCK-8) as well as to CCK agonists were studied. CCK-8 was also infused after CCK antagonists, atropine, hexamethonium or L-nitroarginine. Results show that duodenal response to CCK-8 is excitatory although inhibitory responses can be induced by gastrin. In the jejunum, CCK-8 induces an inhibitory response that is mediated by both CCK-A and -B receptors. Excitatory responses to CCK-8 are due to stimulation of preganglionic receptors while inhibitory responses are NO mediated through stimulation of postganglionic CCK-B receptors. Capsaicin locally applied in duodenal mucosa significantly decreased CCK-8 response, whereas mucosal exposure to lidocaine completely blocked CCK-8 response. In conclusion our results show that CCK response varies along the intestine according to the predominance of excitatory or inhibitory efferent innervation. Moreover, CCK-8 actions are mediated through both extrinsic and intrinsic afferent fibres.
Assuntos
Colecistocinina/metabolismo , Intestino Delgado/inervação , Intestino Delgado/metabolismo , Neurônios Aferentes/metabolismo , Neurônios Eferentes/metabolismo , Anestésicos Locais/farmacologia , Animais , Atropina/farmacologia , Benzodiazepinonas/farmacologia , Devazepida/farmacologia , Duodeno/inervação , Duodeno/metabolismo , Gastrinas/metabolismo , Gastrinas/farmacologia , Hexametônio/farmacologia , Antagonistas de Hormônios/farmacologia , Mucosa Intestinal/metabolismo , Intestino Delgado/efeitos dos fármacos , Jejuno/inervação , Jejuno/metabolismo , Lidocaína/farmacologia , Masculino , Nitroarginina/farmacologia , Oligopeptídeos/farmacologia , Compostos de Fenilureia/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores da Colecistocinina/antagonistas & inibidores , Receptores da Colecistocinina/metabolismo , Sincalida/metabolismo , Sincalida/farmacologia , Tetragastrina/análogos & derivados , Tetragastrina/farmacologiaRESUMO
During the last few years, cholecystokinin (CCK) has emerged as an important hormone. This polypeptide has been located either in peripheral tissues such as the gastro-intestinal tract and the pancreas as well as in the central nervous system. High affinity CCK receptors are divided in two main subtypes: the CCK-A (A for (A for "alimentary") and the CCK-B (B for "brain") receptors. The latters are currently associated with the gastrin receptors. Since CCK is involved in many different biological processes such as gut function, digestive processes, control of feeding behaviour and neurotransmitter release, the therapeutical potential of cholecystokinin receptor ligands seems to be extremely broad and promising. Several families of CCK receptor ligands (peptides, peptidomimetics, peptoids or non-peptides) were prepared during the last twenty years. The main goal of these researches was to improve agonistic or antagonistic potency but also to find selective compounds for a specific CCK receptor subtype. This review presents the recent developments (since 1995) in the chemistry of CCK receptor ligands.
Assuntos
Benzodiazepinas/química , Receptores da Colecistocinina/agonistas , Receptores da Colecistocinina/antagonistas & inibidores , Tetragastrina/análogos & derivados , Animais , Ansiolíticos/química , Ansiolíticos/farmacologia , Benzodiazepinas/farmacologia , Benzodiazepinonas/química , Benzodiazepinonas/farmacologia , Humanos , Indóis/química , Indóis/farmacologia , Cetoácidos/química , Cetoácidos/farmacologia , Ligantes , Meglumina/análogos & derivados , Meglumina/química , Meglumina/farmacologia , Peptoides , Proglumida/análogos & derivados , Proglumida/química , Proglumida/farmacologia , Receptor de Colecistocinina A , Receptor de Colecistocinina B , Compostos de Espiro/química , Compostos de Espiro/farmacologia , Relação Estrutura-AtividadeRESUMO
The neuropeptide substance P is known to have mnemogenic and reinforcing actions and can exert neurotrophic and regenerative effects in vitro as well as in vivo. Furthermore, our previous work in the rat showed that either pre- or post-lesion treatment with substance P can promote functional recovery in cases of partial nigrostriatal dopamine lesions. Other work has provided evidence that the effects of substance P might be differentially encoded by its C- and N-terminal fragments. The C-terminal fragment was found to be reinforcing, whereas the mnemogenic as well as neurotrophic properties have been ascribed to the N-terminal sequences. Given these relations, we asked here whether pre-lesion treatment with either a C- or an N-terminal fragment of substance P might differentially affect the behavioral and neurochemical outcome of nigrostriatal dopamine lesions. Therefore, either substance P1-7 or substance P5-11 (37 nmol/kg each) was administered intraperitoneally daily for eight consecutive days before unilateral 6-hydroxy-dopamine lesions of the substantia nigra. Control rats received pre-lesion treatment with vehicle. Furthermore, we investigated the effects of pre-treatment with Boc-cholecystokinin-4 (0.91 nmol/kg), as we had found an increase in dopamine metabolism in animals that were pre-treated with cholecystokinin-8 in a former study. In accordance with our previous work, drug treatment effects were observed when excluding animals with most severe dopamine lesions: In animals with partial lesions (residual neostriatal dopamine levels of more than 10%), lesion-dependent asymmetries in turning behavior were observed in animals that were pre-treated with vehicle-, substance P1-7, or Boc-cholecysto-kinin-4, whereas turning after pre-treatment with substance P5-11 was not significantly asymmetrical. Furthermore, the ipsi- and contra-lateral neostriatal dopamine levels did not differ significantly in this group. Moreover, pre-treatment with substance P5-11 affected dopamine metabolism in the neostriatum and in the ventral striatum, as indicated by increased ratios of dihydroxyphenyllic acid to dopamine. The data provide the first evidence that the promotive effects of substance-P treatment in the unilateral dopamine lesion model might be mediated by its C-terminal and might depend on actions on residual dopamine mechanisms.
Assuntos
Corpo Estriado/efeitos dos fármacos , Dopamina/metabolismo , Fragmentos de Peptídeos/farmacologia , Substância P/análogos & derivados , Substância P/farmacologia , Substância Negra/efeitos dos fármacos , Tetragastrina/farmacologia , Animais , Corpo Estriado/patologia , Corpo Estriado/fisiologia , Lateralidade Funcional , Masculino , Atividade Motora/efeitos dos fármacos , Oxidopamina/toxicidade , Ácido Pirrolidonocarboxílico/análogos & derivados , Ratos , Ratos Wistar , Substância Negra/patologia , Substância Negra/fisiologia , Tetragastrina/análogos & derivadosRESUMO
This study investigated a role of cholecystokinin (CCK) in the anxiolytic-like action of morphine, an agonist of mu-opioid receptors, in the rat plus-maze model of anxiety. The acute administration of morphine (1 mg/kg) induced a significant increase of exploratory activity in the plus-maze, but did not affect the locomotor activity in the motility test. The higher dose of morphine (2.5 mg/kg) tended to decrease the locomotor activity and, therefore, did not cause the anxiolytic-like action in the plus-maze. The other drugs (naloxone, BOC-CCK-4, L-365,260) and their combinations with morphine (0.5-1 mg/kg) did not affect the locomotor activity of rats. The opioid antagonist naloxone itself (0.5 mg/kg) did not change the exploratory activity in the plus-maze, but potently antagonized the anxiolytic-like action of morphine (1 mg/kg). An agonist of CCK(B)receptors BOC-CCK-4 (1-50 microgram/kg) induced a dose-dependent anxiogenic-like action in the plus-maze. Nevertheless, only one dose of BOC-CCK-4 (10 microgram/kg) completely reversed the action of morphine. Also, one dose of CCK(B)receptor antagonist L-365,260 (10 microgram/kg) was effective to modify the behaviour of rats in the elevated plus-maze. Namely, this dose of L-365,260 increased the ratio between open and total arm entries, a behavioural measure believed to reflect the anxiolytic-like action in the elevated plus-maze. The combination of L-365,260 (100 microgram/kg) with the sub-effective dose of morphine (0.5 mg/kg) caused the anxiolytic-like action in the plus-maze not seen if the drugs were given alone. In conclusion, morphine induces a potent anxiolytic-like action in the elevated plus-maze and CCK is acting as an endogenous antagonist of this effect of morphine.
Assuntos
Ansiolíticos/antagonistas & inibidores , Aprendizagem em Labirinto/efeitos dos fármacos , Morfina/antagonistas & inibidores , Receptores da Colecistocinina/agonistas , Tetragastrina/análogos & derivados , Analgésicos Opioides/antagonistas & inibidores , Analgésicos Opioides/farmacologia , Animais , Ansiolíticos/farmacologia , Ansiedade/tratamento farmacológico , Ansiedade/fisiopatologia , Benzodiazepinonas/farmacologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Interações Medicamentosas , Medo/efeitos dos fármacos , Injeções Subcutâneas , Locomoção/efeitos dos fármacos , Masculino , Morfina/farmacologia , Naloxona/farmacologia , Antagonistas de Entorpecentes/farmacologia , Compostos de Fenilureia/farmacologia , Ratos , Ratos Wistar , Receptor de Colecistocinina B , Tetragastrina/farmacologiaRESUMO
PURPOSE: Three types of acyl tetragastrin (TG), acetyl-TG (C2-TG), butyryl-TG (C4-TG) and caproyl-TG (C6-TG) were synthesized and their in vitro intestinal permeability characteristics were examined using Caco-2 monolayers. METHODS: The disappearance of acyl-TGs from the apical side of Caco-2 monolayers was estimated by analyzing degradation and permeation processes in terms of clearance. RESULTS: The amount of native TG transported to the basolateral side was very low due to its large degradation clearance (CLd) on the apical side. Degradation of TG was reduced by chemical modification with fatty acids, which resulted in an increase in the transport of TG across Caco-2 monolayers. In addition, the permeation clearance (CLp) value of carboxyfluorescein (CF), a paracellular transport and undegradable marker, was increased in the presence of acyl-TGs. Furthermore, we investigated the effects of the protease inhibitors bacitracin and gabexate on the transport of TG across Caco-2 monolayers. In the presence of a low concentration (0.1 mM) of protease inhibitor, the CLd value of TG was reduced, but they did not affect its CLp value. However, a higher concentration (1.0 mM) of bacitracin significantly reduced TG degradation on the apical side, and further increased its CLp value. CONCLUSIONS: We demonstrated that acylation of TG made it resistant to intestinal proteases and caused it to enhance absorption of drugs, including itself, across Caco-2 monolayers. Further, bacitracin acted as both a protease inhibitor and an absorption enhancer.
Assuntos
Absorção Intestinal , Intestino Delgado/metabolismo , Tetragastrina/farmacocinética , Acetilação , Acilação , Transporte Biológico/efeitos dos fármacos , Células CACO-2 , Permeabilidade da Membrana Celular/efeitos dos fármacos , Estabilidade de Medicamentos , Gabexato , Humanos , Intestino Delgado/efeitos dos fármacos , Modelos Biológicos , Inibidores de Serina Proteinase/farmacologia , Tetragastrina/análogos & derivadosRESUMO
This series of experiments examined the effects of the cholecystokinin (CCK) fragments Boc-CCK-4 and CCK-8s on memory, reinforcement and anxiety following unilateral injection into the central nucleus of the amygdala (CeA). In experiment 1, rats with chronically implanted cannulae were injected with CCK-8s or Boc-CCK-4 and were tested on a one-trial uphill avoidance task. Post-trial injection of 20 ng Boc-CCK-4 or 1 ng CCK-8s was found to improve the retention performance, whereas lower and higher doses had no effect. The hypermnestic effects of Boc-CCK-4 and CCK-8s were no longer evident when injection was performed 5 h, rather than immediately, after the learning trial. In experiment 2, the elevated plus-maze was used to gauge anxiogenous properties of intra-amygdala injections of Boc-CCK-4 and CCK-8s in memory-enhancing doses. The treatment with 20 ng Boc-CCK-4 and 1 ng CCK-8s did not influence the number of entries into and time spent on the open and enclosed arms of the maze as well as other anxiety-related behaviors. In experiment 3, possible reinforcing effects of the CCK-fragments were examined. After intra-amygdala injection of Boc-CCK-4 or CCK-8s in memory-enhancing doses the rats were placed into one of four restricted quadrants of a circular open field (closed corral) for a single conditioning trial. Subsequent tests for conditioned corral preference revealed no evidence for reinforcing or aversive effects of the CCK-fragments. In sum, these findings indicate that Boc-CCK-4 and CCK-8s facilitate memory processing upon injection into the CeA without exerting reinforcing or anxiogenous effects.
Assuntos
Tonsila do Cerebelo/efeitos dos fármacos , Ansiedade , Memória/efeitos dos fármacos , Reforço Psicológico , Sincalida/análogos & derivados , Tetragastrina/análogos & derivados , Animais , Aprendizagem da Esquiva/efeitos dos fármacos , Comportamento Animal/efeitos dos fármacos , Condicionamento Psicológico , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Ratos , Ratos Wistar , Sincalida/administração & dosagem , Sincalida/farmacologia , Tetragastrina/administração & dosagem , Tetragastrina/farmacologiaRESUMO
We have previously described the design and development of CI-988, a peptoid analogue of CCK-4 with excellent binding affinity and selectivity for the CCK-B receptor. Due to its anxiolytic profile in animal models of anxiety, this compound was developed as a clinical candidate. However, during its development, it was determined that CI-988 had low bioavailability in both rodent and nonrodent species. In the clinic, it was further established that CI-988 had poor bioavailability. Thus, there was a need to identify an analogue with an improved pharmacokinetic (PK) profile. The poor bioavailability was attributed to poor absorption and efficient hepatic extraction. We envisaged that reducing the molecular weight of the parent compound (5, MW = 614) would lead to better absorption. Thus, we synthesized a series of analogues in which the key alpha-methyltryptophan and adamantyloxycarbonyl moieties, required for receptor binding, were kept intact and the C-terminus was extensively modified. This SAR study led to the identification of tricyclo[3.3.1.1(3,7)]dec-2-yl [1S-[1 alpha(S*)2 beta]-[2-[(2-hydroxycyclohexyl)amino]-1-(1H-indol-3- ylmethyl)-1-methyl-2-oxoethyl]carbamate (CI-1015, 31) with binding affinities of 3.0 and 2900 nM for the CCK-B and CCK-A receptors, respectively. The compound showed CCK-B antagonist profile in the rat ventromedial hypothalamus assay with a Ke of 34 nM. It also showed an anxiolytic like profile orally in a standard anxiety paradigm (X-maze) with a minimum effective dose (MED) of 0.1 microgram/kg. Although the compound is less water soluble than CI-988, oral bioavailability in rat was improved nearly 10 times relative to CI-988 when dosed in HP beta CD. The blood-brain permeability of CI-1015 (31) was also enhanced relative to CI-988 (5). On the basis of the overall improved pharmacokinetic profile as well as enhanced brain penetration, CI-1015 (31) was chosen as a development candidate.
Assuntos
Adamantano/análogos & derivados , Ansiolíticos/síntese química , Receptores da Colecistocinina/antagonistas & inibidores , Tetragastrina/análogos & derivados , Triptofano/análogos & derivados , Adamantano/síntese química , Adamantano/química , Adamantano/farmacocinética , Animais , Ansiolíticos/química , Ansiolíticos/farmacocinética , Ansiolíticos/farmacologia , Disponibilidade Biológica , Barreira Hematoencefálica , Aprendizagem em Labirinto/efeitos dos fármacos , Camundongos , Modelos Moleculares , Estrutura Molecular , Peptoides , Ratos , Ratos Wistar , Receptor de Colecistocinina A , Receptor de Colecistocinina B , Receptores da Colecistocinina/metabolismo , Triptofano/síntese química , Triptofano/química , Triptofano/farmacocinéticaRESUMO
This study investigated the interplay of cholecystokinin (CCK) and endogenous opioid peptides in the regulation of anxiety. The acute administration of non-selective CCK agonist caerulein (1 and 5 microg/kg) and a selective CCK(B) receptor agonist BOC-CCK-4 (1, 10 and 50 microg/kg) induced a dose-dependent anxiogenic-like action in the plus-maze model of anxiety. BOC-CCK-4 displayed a similar efficacy with caerulein, indicating that the described effect was mediated via CCK(B) receptor subtype. The opioid antagonist naloxone itself (0.5 mg/kg) did not change the exploratory activity of rats in the plus-maze. However, the combination of naloxone with the sub-effective doses of caerulein (1 microg/kg) and BOC-CCK-4 (1 microg/kg) induced a significant inhibition of exploratory behaviour in rats. Accordingly, CCK and endogenous opioid peptides have an antagonistic role in the exploratory model of anxiety in rats.
Assuntos
Ansiedade/induzido quimicamente , Colecistocinina/agonistas , Naloxona/farmacologia , Antagonistas de Entorpecentes/farmacologia , Animais , Ceruletídeo/farmacologia , Modelos Animais de Doenças , Sinergismo Farmacológico , Comportamento Exploratório/efeitos dos fármacos , Masculino , Ratos , Ratos Wistar , Receptor de Colecistocinina B , Receptores da Colecistocinina/agonistas , Receptores da Colecistocinina/fisiologia , Tetragastrina/análogos & derivados , Tetragastrina/farmacologiaRESUMO
The present study investigated the effects of agonists of the neuropeptide cholecystokinin (CCK) on neostriatal neurons in order to confirm the existence of CCK-A receptors in the rat caudate-putamen. Single unit activity was recorded in rats anaesthetized with urethane. The CCK-A receptor agonists A-71378 (desamino-Tyr(SO3H)-Nle-Gly-Trp-Nle-(N-methyl)Asp-Phe-NH2), and A-71623 (Boc-Trp-Lys(epsilon-N-2-methylphenylamino-carbonyl)-Asp-(N-methyl )Phe-NH2, as well as the CCK-B receptor agonist Suc-CCK-4 (Suc-Trp-(N-methyl)Nle-Asp-Phe-NH2) were iontophoretically administered with multibarrel capillaries. About one-third of the neurons responded to the CCK-A receptor agonists. Excitatory effects predominated. The CCK-B receptor agonist also induced mainly increases in the neuronal firing. Several neurons were affected by both types of agonists. The results suggest that in the rat neostriatum not only CCK-B receptors, but also CCK-A receptors seem to mediate the effects of cholecystokinin.
Assuntos
Neostriado/citologia , Neurônios/química , Receptores da Colecistocinina/agonistas , Animais , Eletrofisiologia , Masculino , Neurônios/efeitos dos fármacos , Oligopeptídeos/farmacologia , Ratos , Ratos Wistar , Receptor de Colecistocinina A , Receptor de Colecistocinina B , Tetragastrina/análogos & derivados , Tetragastrina/farmacologiaRESUMO
We previously reported novel Boc-CCK-4 (Boc-Trp-Met-Asp-Phe-NH2) derivatives possessing the general structure Boc-Trp-Lys[N epsilon-CO-NH-(R-Ph)]-Asp-Phe-NH2 (Shiosaki et al. J. Med. Chem. 1991, 34, 2837-2842). In contrast to Boc-CCK-4, which is 70-fold selective for the CCK-B receptor, the modified lysine-bearing tetrapeptides were highly potent and selective full agonists at the CCK-A receptor. Further investigation of the structure-activity profile following modification of the substituted phenylurea moiety appended off the lysine revealed that moving certain substituents, e.g. nitro or acetyl, from the 2- or 3-position on the phenyl ring to the 4-position, a relatively minor and subtle structural modification within the tetrapeptide, resulted in loss of CCK-A receptor selectivity and development of a trend toward CCK-B selectivity. These tetrapeptides, e.g. Boc-Trp-Lys[N epsilon-CO-NH-(4-NO2-Ph)]-Asp-Phe-NH2 and Boc-Trp-Lys[N epsilon-CO-NH-(4-Ac-Ph)]-Asp-Phe-NH2, were full agonists relative to CCK-8 in stimulating intracellular calcium mobilization in a cell line that expresses the CCK-B receptor.