RESUMO
INTRODUCTION: Depression, anxiety, and/or panic disorder are often comorbid and have a complex etiology mediated through the same neuronal network. Cholecystokinin-tetrapeptide (CCK-4), a synthetic analog of the endogenous neuropeptide cholecystokinin (CCK), is thought to be implicated in this network. The CCK-4 challenge model is an accepted method of investigating the pathophysiology of panic and has been shown to mediate neuronal activation via the transient receptor potential canonical (TRPC) ion channels. OBJECTIVES: This study aimed to assess the pharmacodynamic effects of BI 1358894, a small-molecule inhibitor of TRPC ion channel members 4 and 5 (TRPC4/5), on CCK-4-induced anxiety/panic-like symptoms and evaluate circuit engagement. METHODS: Twenty healthy male CCK-4-sensitive volunteers entered a Phase I, double blind, randomized, two-way cross-over, single dose, placebo-controlled trial. Randomization was to oral BI 1358894 100 mg in the fed state followed by oral placebo in the fed state, or vice versa. Treatments were administered 5 h prior to intravenous CCK-4 50 µg. The primary endpoint was maximum change from baseline of the Panic Symptom Scale (PSS) sum intensity score after CCK-4 injection. Further endpoints included the emotional faces visual analog score (EVAS), the Spielberger State-Trait Anxiety Inventory (STAI), plasma adrenocorticotropic hormone (ACTH), and serum cortisol values. The safety and tolerability of BI 1358894 was assessed based on a number of parameters including occurrence of adverse events (AEs). All pharmacodynamic, pharmacokinetic, and safety endpoints were analyzed using descriptive statistics. RESULTS: Single oral doses of BI 1358894 were generally well tolerated by the healthy male volunteers included in this study. Adjusted mean maximum change from baseline in PSS sum intensity score was 24.4 % lower in volunteers treated with BI 1358894 versus placebo, while adjusted mean maximum change from baseline of EVAS was reduced by 19.2 % (BI 1358894 vs placebo). The STAI total score before CCK-4 injection was similar in both groups (placebo: 25.1; BI 1358894: 24.3). Relative to placebo, BI 1358894 reduced CCK-4-induced mean maximum plasma ACTH and serum cortisol values by 58.6 % and 27.3 %, respectively. Investigator-assessed drug-related AEs were reported for 13/20 participants (65.0 %). There were no serious or severe AEs, AEs of special interest, AEs leading to discontinuation of trial medication, or deaths. CONCLUSIONS: Overall, BI 1358894 reduced psychological and physiological responses to CCK-4 compared with placebo, as measured by PSS, subjective EVAS and objectively measured stress biomarkers. BI 1358894 had a positive safety profile, and single oral doses were well tolerated by the healthy volunteers. This trial (NCT03904576/1402-0005) was registered on Clinicaltrials.gov on 05.04.19.
Assuntos
Hidrocortisona , Tetragastrina , Humanos , Masculino , Tetragastrina/efeitos adversos , Hormônio Adrenocorticotrópico , Ansiedade/tratamento farmacológico , Método Duplo-Cego , BiomarcadoresRESUMO
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
The widespread involvement of the cholecystokinin-2/gastrin receptor (CCK2R) in multiple (patho)physiological processes has propelled extensive searches for nonpeptide small-molecule CCK2R antagonists. For the past three decades, considerable research has yielded numerous chemically heterogeneous compounds. None of these entered into the clinic, mainly because of inadequate biological effects. However, it appears that the ultimate goal of a clinically useful CCK2R antagonist is now just around the corner, with the most promising compounds, netazepide and nastorazepide, now in Phase II clinical trials. Here, we illustrate the structure-activity relationships (SARs) of stablished CCK2R antagonists of various structural classes, and the most recent proof-of-concept studies where new applicabilities of CCK2R antagonists as visualizing agents are presented.
Assuntos
Receptor de Colecistocinina B/antagonistas & inibidores , Animais , Benzodiazepinonas/química , Benzodiazepinonas/uso terapêutico , Ensaios Clínicos como Assunto , Furanos/química , Furanos/uso terapêutico , Humanos , Lactamas/química , Lactamas/uso terapêutico , Receptor de Colecistocinina B/metabolismo , Relação Estrutura-Atividade , Sulfonamidas/química , Sulfonamidas/uso terapêutico , Tetragastrina/química , Tetragastrina/uso terapêuticoRESUMO
Copeptin, the C-terminal part of the hypothalamic arginine vaspopressin (AVP) precursor, closely mirrors the production of AVP and was proposed as an easily measured novel marker of the individual stress level in man. First data in male volunteers proposed copeptin as a potential endocrine surrogate marker of cholecystokinin-tetrapeptide (CCK-4)-induced panic. We tried to replicate these pilot data and to extend them to the other sex. 46 healthy human subjects (29 men, 17 women) were given an intravenous bolus of 50⯵g CCK-4. Basal and stimulated plasma copeptin was measured and panic symptoms were assessed using the Acute Panic Inventory (API). Basal copeptin was significantly lower in women vs. men, while men showed a significantly higher CCK-4-induced increase of copeptin. In contrast, female subjects displayed a signifcantly higher increase of API ratings by CCK-4. No significant correlations of panic symptoms and copeptin release induced by CCK-4 could be found, neither in man, nor in women, nor in the total sample. A sexual dimorphism in copeptin secretion and in panic response was demonstrated. Prior unexpected findings of copeptin release as an objective read-out of panic could not be replicated. The role of the vasopressinergic system in panic anxiety needs further study in panic patients and in healthy man, using also other panic provocation paradigms.
Assuntos
Glicopeptídeos/sangue , Transtorno de Pânico/induzido quimicamente , Transtorno de Pânico/diagnóstico , Pânico/efeitos dos fármacos , Caracteres Sexuais , Tetragastrina/efeitos adversos , Adaptação Psicológica/efeitos dos fármacos , Adulto , Biomarcadores/sangue , Feminino , Glicopeptídeos/metabolismo , Voluntários Saudáveis , Humanos , Masculino , Transtorno de Pânico/sangue , Via Secretória/efeitos dos fármacos , Adulto JovemRESUMO
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
Endogenous cholecystokinin tetrapeptide (CCK-4, Trp-Met-Asp-Phe-NH2) is a fragment derived from a larger peptide hormone, cholecystokinin (or gastrin). As a panicogenic agent, CCK-4 is commonly used in clinic settings to induce panic attacks for the study of new anxiolytic drugs. However, few studies on CCK-4 metabolism have been published to date. In the present study, we investigate the metabolism of CCK-4 in liver microsomes of human (HLM), Rhesus Monkey (RMLM), Sprague-Dawley rat (RLM) and CD1 mouse (MLM) using ultra-high performance liquid chromatography coupled to a high resolution mass spetrometer. Ten metabolites, inlcuding tryptophan (M1), tryptophan amide (M2), hydroxy metabolites (M3-M5), truncated peptides (M6-M9), and CCK-4 acid (M10), were identified and 8 of them were reported for the first time. The metabolic pattern of CCK-4 in HLM was distinctly different from these in RMLM, RLM, and MLM. M2 and M9 were the major metabolites in HLM and accounted for 19.8% and 13.4% of initial CCK-4, respectively. In contrast, M2 was the major metabolite in RMLM and accounted for 41.4%, whereas M6 was the major metabolite in RLM and account for 39.1%. Three major metabolites M2, M7 and M8 in MLM accounted for 22.6%, 17.9% and 17.8% of initial CCK-4, respectively. Chemical inhibition experiment showed that aminopeptidase and/or endopeptidase hydrolysis were the major metabolic pathways in human to generate these metabolites. We further showed that cytochrome P450 were also involved in the metabolism of CCK-4 via hydroxylation, but to a less extend. These findings provide valuable information for the metabolic processes of CCK-4 among various species and an important reference basis for its safety evaluation and rational clinical application.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Espectrometria de Massas/métodos , Microssomos Hepáticos/metabolismo , Tetragastrina/metabolismo , Animais , Humanos , Hidroxilação , Macaca mulatta , Camundongos , Ratos , Ratos Sprague-DawleyRESUMO
Intravenous cholecystokinin-tetrapeptide (CCK-4) administration reliably and dose-dependently provokes panic anxiety in man, accompanied by adrenocorticotropic hormone (ACTH) and cortisol release. Preclinical findings suggest that behavioral and endocrine effects of CCK-4 are mediated via corticotropin-releasing hormone (CRH) release. Anxiogenic stimulation of the central CCK-receptors in man was shown to increase as well vasopressin (AVP), which acts synergistically with CRH as pituitary-adrenocortical axis stimulator during stress. Copeptin (CoP), the C-terminal part of pre-pro-AVP, is released in an equimolar ratio to AVP. It is more stable in the circulation and easier to determine than AVP and it was found to closely mirror the production of AVP. So far, CoP secretion has not been characterized during panic provocation. In 30 healthy male human subjects, we repeatedly measured CoP in plasma during a panic challenge and studied its correlation to Acute Panic Inventory (API) ratings and plasma ACTH and cortisol. CoP levels correlated positively with the increase in API ratings (r=0.41, p=0.03), while ACTH or cortisol did not (r=0.08, p=0.68 and r=0.12, p=0.53, respectively). CoP levels correlated also positively with ACTH (r=0.48, p=0.009) and cortisol (r=0.48, p=0.01) concentrations throughout the CCK-4 challenge. As expected, we found a positive correlation between plasma ACTH and cortisol levels (r=0.57, p=0.001). A vasopressinergic activation during CCK-4 induced panic was demonstrated, which was correlated positively to panic symptoms and pituitary-adrenocortical release. Our findings suggest a role of CoP as a potential surrogate marker of CCK-4 panic symptoms. Further studies are needed to replicate our results and to further clarify the role of CoP as a stress-sensitive hormone in different panic paradigms as well as in panic patients.
Assuntos
Hormônio Adrenocorticotrópico/sangue , Ansiedade/induzido quimicamente , Glicopeptídeos/sangue , Hidrocortisona/sangue , Neurotransmissores/farmacologia , Pânico , Receptores da Colecistocinina/agonistas , Tetragastrina/farmacologia , Adulto , Biomarcadores/sangue , Humanos , Masculino , Pânico/fisiologia , Adulto JovemRESUMO
Group II metabotropic glutamate receptors (mGluR2/3) are suggested to modulate anxiety, arousal, and stress including autonomic control. However, no study has investigated mGluR2/3-related effects on baseline autonomic activity and reactivity to emotional challenge in humans as yet. Using a double-blind, randomized placebo-controlled, cross-over study design, we investigated the influence of a 1-week treatment with the mGluR2/3 agonist LY544344, prodrug of LY354740, on autonomic reactivity to a cholecystokinin tetrapeptide (CCK-4) panic challenge in eight healthy young men. The main outcome measures were time and frequency domain heart rate variability parameters during baseline, CCK-4 challenge, and recovery. There was no evidence for LY544344-mediated effects on baseline and CCK-4 challenge vagal activity, but a significantly lower recovery low frequency (%) and low frequency/high frequency ratio in the LY544344 group, suggesting enhanced autonomic recovery. This pilot study provides first human data indicating that mGluR2/3 agonism is involved in autonomic responsiveness, suggesting an important role of mGluR2/3 in central autonomic regulation.
Assuntos
Alanina/análogos & derivados , Compostos Bicíclicos com Pontes/farmacologia , Frequência Cardíaca/efeitos dos fármacos , Pânico/efeitos dos fármacos , Receptores de Glutamato Metabotrópico/agonistas , Tetragastrina/farmacologia , Adulto , Alanina/farmacologia , Estudos Cross-Over , Método Duplo-Cego , Voluntários Saudáveis , Humanos , Masculino , Projetos Piloto , Adulto JovemRESUMO
An excitatory-inhibitory neurotransmitter dysbalance has been suggested in pathogenesis of panic disorder. The neuropeptide S (NPS) system has been implicated in modulating GABA and glutamate neurotransmission in animal models and to genetically drive altered fear circuit function and an increased risk of panic disorder in humans. Probing a multi-level imaging genetic risk model of panic, in the present magnetic resonance spectroscopy (MRS) study brain glutamate+glutamine (Glx) levels in the bilateral anterior cingulate cortex (ACC) during a pharmacological cholecystokinin tetrapeptide (CCK-4) panic challenge were assessed depending on the functional neuropeptide S receptor gene (NPSR1) rs324981 A/T variant in a final sample of 35 healthy male subjects. The subjective panic response (Panic Symptom Scale; PSS) as well as cortisol and ACTH levels were ascertained throughout the experiment. CCK-4 injection was followed by a strong panic response. A significant time×genotype interaction was detected (p=.008), with significantly lower ACC Glx/Cr levels in T allele carriers as compared to AA homozygotes 5min after injection (p=.003). CCK-4 induced significant HPA axis stimulation, but no effect of genotype was discerned. The present pilot data suggests NPSR1 gene variation to modulate Glx levels in the ACC during acute states of stress and anxiety, with blunted, i.e. possibly maladaptive ACC glutamatergic reactivity in T risk allele carriers. Our results underline the notion of a genetically driven rapid and dynamic response mechanism in the neural regulation of human anxiety and further strengthen the emerging role of the NPS system in anxiety.
Assuntos
Ácido Glutâmico/metabolismo , Glutamina/metabolismo , Giro do Cíngulo/metabolismo , Pânico/fisiologia , Receptores Acoplados a Proteínas G/genética , Hormônio Adrenocorticotrópico/sangue , Adulto , Análise Química do Sangue , Creatina/metabolismo , Ensaio de Imunoadsorção Enzimática , Genótipo , Técnicas de Genotipagem , Humanos , Hidrocortisona/sangue , Masculino , Projetos Piloto , Espectroscopia de Prótons por Ressonância Magnética , Escalas de Graduação Psiquiátrica , Tetragastrina , Fatores de Tempo , Adulto JovemRESUMO
BACKGROUND: Central serotonergic pathways influence brain areas involved in vagal cardiovascular regulation and, thereby, influence sympathetic efferent activity. Selective serotonin reuptake inhibitors (SSRIs) affect multiple serotonergic pathways, including central autonomic pathways. However, only a few studies have assessed SSRI-mediated effects on autonomic reactivity in healthy individuals using heart rate variability (HRV). METHODS: The present study assessed the influence of long-term treatment with escitalopram (ESC) on autonomic reactivity to an intravenous application of 50 µg cholecystokinin tetrapeptide (CCK-4) in 30 healthy young men using a double-blind, placebo (PLA)-controlled, randomized, within-subject cross-over design. Main outcome measures were time- and frequency-domain HRV parameters, assessed at both baseline and immediately after CCK-4 application. RESULTS: Results showed substantial effects for the treatment × CCK-4 challenge interaction with respect to heart rate (p < 0.001; pη(2) = 0.499), SDNN (p < 0.001; pη(2) = 576), RMSSD (p = 0.015; pη(2) = 194), NN50% (p = 0.008; pη(2) = 0.224), and LF% (p = 0.014; pη(2) = 0.196), and moderate effects with respect HF% (p = 0.099; pη(2) = 0.094), with PLA subjects showing a higher increase in HR and SDNN and a higher decrease in RMSSD, NN50, LF and HF than subjects in the ESC condition. Thus, ESC treatment significantly blunted the autonomic reactivity to CCK-4. Secondary analysis indicated no effect of the 5-HTTLPR polymorphism on CCK-4-induced autonomic response. CONCLUSIONS: Our results support findings suggesting an effect of SSRI treatment on autonomic regulation and provide evidence that ESC treatment is associated with blunted autonomic reactivity in healthy men.
Assuntos
Sistema Nervoso Autônomo/efeitos dos fármacos , Citalopram/farmacologia , Frequência Cardíaca/efeitos dos fármacos , Inibidores Seletivos de Recaptação de Serotonina/farmacologia , Administração Intravenosa , Adulto , Sistema Nervoso Autônomo/fisiologia , Citalopram/administração & dosagem , Estudos Cross-Over , Método Duplo-Cego , Hormônios Gastrointestinais/administração & dosagem , Hormônios Gastrointestinais/farmacologia , Voluntários Saudáveis , Humanos , Masculino , Distribuição Aleatória , Inibidores Seletivos de Recaptação de Serotonina/administração & dosagem , Tetragastrina/administração & dosagem , Tetragastrina/farmacologia , Fatores de Tempo , Resultado do Tratamento , Adulto JovemRESUMO
Novel strategies are needed to expedite the generation and optimization of peptide probes targeting G protein-coupled receptors (GPCRs). We have previously shown that membrane tethered ligands (MTLs), recombinant proteins comprised of a membrane anchor, an extracellular linker, and a peptide ligand can be used to identify targeted receptor modulators. Although MTLs provide a useful tool to identify and/or modify functionally active peptides, a major limitation of this strategy is the reliance on recombinant protein expression. We now report the generation and pharmacological characterization of prototype peptide-linker-lipid conjugates, synthetic membrane anchored ligands (SMALs), which are designed as mimics of corresponding MTLs. In this study, we systematically compare the activity of selected peptides as MTLs versus SMALs. As prototypes, we focused on the precursor proteins of mature Substance P (SubP) and Cholecystokinin 4 (CCK4), specifically non-amidated SubP (SubP-COOH) and glycine extended CCK4 (CCK4-Gly-COOH). As low affinity soluble peptides these ligands each presented a challenging test case for assessment of MTL/SMAL technology. For each ligand, MTLs and corresponding SMALs showed agonist activity and comparable subtype selectivity. In addition, our results illustrate that membrane anchoring increases ligand potency. Furthermore, both MTL and SMAL induced signaling can be blocked by specific non-peptide antagonists suggesting that the anchored constructs may be orthosteric agonists. In conclusion, MTLs offer a streamlined approach for identifying low activity peptides which can be readily converted to higher potency SMALs. The ability to recapitulate MTL activity with SMALs extends the utility of anchored peptides as probes of GPCR function.
Assuntos
Peptídeos/química , Receptores Acoplados a Proteínas G/química , Glicina/química , Células HEK293 , Humanos , Ligantes , Piperidinas/química , Plasmídeos/metabolismo , Estrutura Terciária de Proteína , Receptores da Neurocinina-1/química , Proteínas Recombinantes/química , Transdução de Sinais , Substância P/química , Tetragastrina/químicaRESUMO
We investigated the involvement of neuropeptide Y (NPY) in the modulation of cholecystokinin-4 (CCK-4)-evoked anxiety and depression. Adult male mice were injected with vehicle, CCK-4, NPY, NPY Y1 receptor agonist [Leu(31), Pro(34)]-NPY or antagonist BIBP3226, via intracerebroventricular route, and subjected to social interaction or forced swim test (FST) for the evaluation of anxiety- and depression-like phenotypes, respectively. To assess the interactions between the two systems, if any, NPYergic agents were administered prior to CCK-4 and the animals were subjected to these behavioral tests. Treatment with CCK-4 or BIBP3226 dose-dependently reduced social interaction time, while NPY or [Leu(31), Pro(34)]-NPY produced opposite effect. CCK-4 treatment increased immobility time in FST. This effect was reversed by NPY and [Leu(31), Pro(34)]-NPY, although BIBP3226 per se did not alter the immobility time. In a combination study, the anxiogenic or depressive effects of CCK-4 were attenuated by NPY or [Leu(31), Pro(34)]-NPY and potentiated by BIBP3226. The brains of CCK-4 treated rats were processed for NPY immunohistochemistry. Following CCK-4 treatment, the nucleus accumbens shell (AcbSh), ventral part of lateral division of the bed nucleus of stria terminalis (BSTLV), hypothalamic paraventricular nucleus and locus coeruleus showed a reduction in NPY-immunoreactive fibers. Population of NPY-immunopositive cells was also decreased in the AcbSh, BSTLV, prefrontal cortex and hypothalamic arcuate nucleus (ARC). However, NPY-immunoreaction in the fibers of the ARC and cells of the central nucleus of amygdala was unchanged. We conclude that, inhibition of NPY signaling in the brain by CCK-4 might be causal to anxiety- and depression-like behaviors.
Assuntos
Ansiedade/fisiopatologia , Encéfalo/fisiopatologia , Depressão/fisiopatologia , Neuropeptídeo Y/metabolismo , Receptores de Neuropeptídeo Y/metabolismo , Tetragastrina/metabolismo , Animais , Ansiolíticos/farmacologia , Ansiedade/tratamento farmacológico , Arginina/análogos & derivados , Arginina/farmacologia , Encéfalo/efeitos dos fármacos , Depressão/tratamento farmacológico , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Imuno-Histoquímica , Masculino , Camundongos , Atividade Motora/efeitos dos fármacos , Atividade Motora/fisiologia , Neuropeptídeo Y/administração & dosagem , Testes Neuropsicológicos , Psicotrópicos/administração & dosagem , Receptores de Neuropeptídeo Y/agonistas , Receptores de Neuropeptídeo Y/antagonistas & inibidores , Comportamento Social , Natação , Tetragastrina/administração & dosagemRESUMO
Abnormal serotonin transporter (5-HTT) function and autonomic nervous system (ANS) dysregulation has been proposed in panic disorder. However, in contrast to hypothalamo-pituitary-adrenocortical (HPA) functioning, ANS reactivity during panic response has yet not been investigated in humans with respect to the 5-HTT genotype. The present study assessed the influence of challenging by cholecystokinin tetrapeptide (CCK-4) on heart rate variability (HRV) measures, to monitor autonomic reactivity and its relationship to 5-HTT-linked polymorphic region (5-HTTLPR) genotypes. We hypothesized substantial effects of the 5-HTTLPR genotype on autonomic reactivity. We studied 30 healthy young men, 15 of each with the long/long (l/l) or short/short (s/s) genotype for the 5-HTTLPR. All participants received an intravenous application of 50 µg CCK-4. HRV measures were assessed in both groups at baseline and immediately after CCK-4 application. Our results indicated lower parasympathetic activity in s/s carriers during baseline, time and frequency domain measures. CCK-4 application significantly enhanced the sympathetic tone in both groups, leading to diminished group differences. A significant treatment by genotype effect indicated reduced autonomic reactivity to CCK-4 challenge in the s/s compared to l/l carriers. Our findings show enhanced sympathetic and/or diminished cardiac vagal activity under basal conditions and blunted autonomic reactivity in s/s vs. l/l carriers. Our study provides novel data supporting claims that the s/s genotype represents a genetic vulnerability factor associated with inadequate hyporeactivity to stress and extends current knowledge on the impact of the central serotonergic activity on the sympathoadrenal pathway.
Assuntos
Fármacos do Sistema Nervoso Autônomo/farmacologia , Frequência Cardíaca/efeitos dos fármacos , Frequência Cardíaca/genética , Proteínas da Membrana Plasmática de Transporte de Serotonina/genética , Tetragastrina/farmacologia , Análise de Variância , Eletrocardiografia , Genótipo , Humanos , Masculino , PânicoRESUMO
The anxiolytic effects of GB-115, a retroanalogue of cholecystokinin-4, administered orally to outbred and inbred animals with different level of emotionality, were studied in the open field test and elevated plus-maze test. The anxiolytic effect of talanax was observed in outbred mice (0.1-0.5 mg/kg) and in inbred BALB/c mice (0.1 and 5.0 mg/kg) in the open field test. GB-115 increased the time of entries into open arms in outbred rats (0.5-0.7 mg/kg) and in BALB/c mice (0.1 mg/kg). These data confirmed the dependence of GB-115 effect on the phenotype of emotional stress response and demonstrated a shift of anxiolytic doses of the preparation from 0.006-0.100 mg/kg in intraperitoneal administration to 0.1-5.0 mg/kg in oral treatment.
Assuntos
Ansiolíticos/farmacologia , Colecistocinina/farmacologia , Dipeptídeos/farmacologia , Estresse Psicológico/tratamento farmacológico , Tetragastrina/farmacologia , Administração Oral , Animais , Comportamento Animal/efeitos dos fármacos , Colecistocinina/análogos & derivados , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , RatosRESUMO
According to preclinical studies, glutamate has been implicated in the pathogenesis of anxiety. In order to elucidate the role of glutamate in anxiety and panic in humans, brain glutamate+glutamine (Glx) levels were measured during cholecystokinin-tetrapeptide (CCK-4)-induced panic using magnetic resonance spectroscopy (MRS). Eighteen healthy subjects underwent a CCK-4 challenge. MR spectra were obtained from the anterior cingulate cortex (ACC) using a single voxel point-resolved spectroscopy method and analyzed using LCModel. A combined fitting of Glx was performed. Panic was assessed using the Acute Panic Inventory (API) and Panic Symptom Scale (PSS) scores. Moreover, hypothalamic-pituitary-adrenal axis stimulation was monitored throughout the challenge. There was a significant panic response following CCK-4 as revealed by a marked increase in both the panic scores (API: F(1,17)=149.41; p<0.0001; PSS: F(1,17)=88.03; p<0.0001) and heart rate (HR: F(1,17)=72.79; p<0.0001). MRS measures showed a significant increase of brain Glx/creatine (Glx/Cr) levels peaking at 2-10 min after challenge (F(1,17)=15.94; p=0.001). There was also a significant increase in CCK-4-related cortisol release (F(6,11)=8.68; p=0.002). Finally, significant positive correlations were found between baseline Glx/Cr and both APImax (r=0.598; p=0.009) and maximum heart rate (HR(max)) during challenge (r=0.519; p=0.027). Our results suggest that CCK-4-induced panic is accompanied by a significant glutamate increase in the bilateral ACC. The results add to the hypothesis of a disturbance of the inhibitory-excitatory equilibrium and suggest that apart from static alterations rapid and dynamic neurochemical changes might also be relevant for the neural control of panic attacks.
Assuntos
Ácido Glutâmico/metabolismo , Transtorno de Pânico/metabolismo , Hormônio Adrenocorticotrópico/sangue , Adulto , Creatina/metabolismo , Neuroimagem Funcional , Glutamina/metabolismo , Giro do Cíngulo/metabolismo , Frequência Cardíaca , Humanos , Hidrocortisona/sangue , Sistema Hipotálamo-Hipofisário/metabolismo , Masculino , Transtorno de Pânico/sangue , Transtorno de Pânico/induzido quimicamente , Sistema Hipófise-Suprarrenal/metabolismo , Escalas de Graduação Psiquiátrica , TetragastrinaRESUMO
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
BACKGROUND: Benzodiazepines modulate γ-aminobutyric acid type A (GABA(A)) receptors throughout the brain. However, it is not fully understood which brain regions within anxiety-related brain circuits are really responsible for their anxiolytic effects and how these regions interact. METHODS: We investigated whether the benzodiazepine alprazolam affects activity in distinct brain regions within anxiety-related circuits during an experimental anxiety paradigm by means of functional magnetic resonance imaging (fMRI). Panic symptoms were elicited by a bolus injection of the neuropeptide cholecystokinin-tetrapeptide (CCK-4) in 16 healthy male subjects in a double-blind, placebo-controlled design. Functional brain activation patterns were determined before and during the CCK-4-challenge without pretreatment and after treatment with either placebo or 1 mg alprazolam. RESULTS: The CCK-4 induced anxiety and elicited widely distributed activation patterns in anxiety-related brain circuits, especially in the rostral anterior cingulate cortex (rACC), which was attenuated after alprazolam treatment. In contrast to placebo, alprazolam abolished the activation of the rACC after challenge with CCK-4 (p<.005, corrected for multiple comparisons) and increased functional connectivity between the rACC and other anxiety-related brain regions such as amygdala and prefrontal cortex. Moreover, the reduction in the CCK-4 induced activation of the rACC correlated with the anxiolytic effect of alprazolam (r(p) = .52; p = .04). CONCLUSIONS: These findings put forward the rACC as a target for benzodiazepines and suggest that the CCK-4/fMRI paradigm might represent a human translational model for the investigation of anxiolytic drugs.
Assuntos
Alprazolam/farmacologia , Ansiolíticos/farmacologia , Ansiedade/tratamento farmacológico , Moduladores GABAérgicos/farmacologia , Giro do Cíngulo/efeitos dos fármacos , Adulto , Alprazolam/uso terapêutico , Ansiolíticos/uso terapêutico , Ansiedade/induzido quimicamente , Ansiedade/fisiopatologia , Encéfalo/efeitos dos fármacos , Encéfalo/fisiopatologia , Mapeamento Encefálico , Método Duplo-Cego , Moduladores GABAérgicos/uso terapêutico , Giro do Cíngulo/fisiopatologia , Humanos , Processamento de Imagem Assistida por Computador , Imageamento por Ressonância Magnética , Masculino , TetragastrinaRESUMO
This study investigated the roles of cholecystokinin (CCK)(A) and CCK(B) receptors on CCK-4-induced anxiety-like behaviors in mice through behavioral and neural evaluations. Anxiety-like behaviors in mice were induced by an intracerebroventricular (i.c.v.) administration of CCK-4, which can bind to both CCK(A) and CCK(B) receptors. The effects of CCK(A) and CCK(B) receptor antagonists (devazepide and CI-988, respectively) were examined using mouse anxiety tests (elevated-plus maze and light-dark box) and also by examining neuronal activities through EEG monitoring and c-Fos immunohistochemistry in the cortex and amygdala. CCK-4 (3 µg/kg of body weight i.c.v.) significantly induced mouse anxiety-like behaviors in the anxiety tests and also affected their EEG patterns with respect to pre-drug tracing, resulting in increase in spectral power in relative power distribution in the delta and theta bands (0.5-5 Hz frequency bands) and also in increase in c-Fos immunopositive neuron counts. These CCK-4 effects were completely suppressed by 1.0mg/kg CCK(B) receptor antagonist, CI-988, while the same amount of CCK(A) receptor antagonist, devazepide was partly able to suppress the same effects. These findings indicated that not only CCK(B) receptors but also CCK(A) receptors in the brain play important roles in regulating anxiety-like behaviors in mice. The present study also proposed a possibility that cortical EEG is useful for assessing anxiety.
Assuntos
Ansiedade/induzido quimicamente , Comportamento Animal/efeitos dos fármacos , Ondas Encefálicas/fisiologia , Encéfalo/metabolismo , Receptor de Colecistocinina A/metabolismo , Receptor de Colecistocinina B/metabolismo , Tetragastrina/toxicidade , Adaptação Fisiológica/efeitos dos fármacos , Análise de Variância , Animais , Ansiedade/patologia , Ansiedade/fisiopatologia , Encéfalo/efeitos dos fármacos , Encéfalo/fisiopatologia , Mapeamento Encefálico , Ondas Encefálicas/efeitos dos fármacos , Devazepida/farmacologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Eletroencefalografia , Regulação da Expressão Gênica/efeitos dos fármacos , Indóis/farmacologia , Injeções Intraventriculares , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Meglumina/análogos & derivados , Meglumina/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Receptor de Colecistocinina A/antagonistas & inibidores , Receptor de Colecistocinina B/antagonistas & inibidores , Análise EspectralRESUMO
Cholecystokinin-tetrapeptide (CCK-4)-induced panic attacks are reportedly attenuated by effective treatment with antipanic antidepressants in patients with panic disorder, but in healthy volunteers such effects are not well studied. The aim of this study was to assess the effect of 6-week treatment with an SSRI escitalopram on CCK-4-induced symptoms in healthy volunteers, who previously responded with a panic attack to CCK-4 challenge. A total of 18 healthy subjects (10 males and eight females, mean age 22.5 ± 5.8) received a 6-week treatment with escitalopram (10 mg/day) and placebo followed by CCK-4 challenge (50 µg) in a double-blind crossover design. The panic rate was 67% after treatment with escitalopram and 56% after treatment with placebo (p = 0.7). Thus, the results showed a significant reduction in CCK-4-induced panic rates without significant differences between escitalopram and placebo conditions. There were no significant effects of either treatment on any other variable of anxiety or cardiovascular indices. Secondary analysis showed no effect of gender or 5-HTTLPR polymorphism on response to CCK-4 challenge. This study demonstrated that in contrast to the findings in patients with panic disorder, in CCK-4-sensitive healthy volunteers the treatment with an antipanic SSRI did not cause a reduction of CCK-4-induced panic attacks beyond the effect of placebo. The mechanisms behind this discrepancy and the reasons of the decrease in sensitivity to CCK-4 challenge on repeated administration remain to be clarified in future studies.