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1.
Cell Physiol Biochem ; 53(4): 701-712, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31592599

RESUMO

BACKGROUND/AIMS: Cholinergic signalling mediated by the activation of muscarinic and nicotinic receptors has been described in the literature as a classic and important signalling pathway in the regulation of the inflammatory response. Recent research has investigated the role of acetylcholine, the physiological agonist of these receptors, in the control of energy homeostasis at the central level. Studies have shown that mice that do not express acetylcholine in brain regions regulating energy homeostasis present with excessive weight gain and hyperphagia. However, it has not yet been well-described in the literature which cholinergic receptor subunits are involved in this response; moreover, the signalling pathways responsible for the observed effects are not fully delineated. The hypothalamus is the regulating centre of energy homeostasis, and the α7 subunit of the nicotinic acetylcholine receptor (α7nAChR) is highly expressed in this region. When active, α7nAChR recruits proteins such as JAK2/STAT3 to mediate its signalling; the same intracellular components are required by leptin, an anorexigenic hormone. The aim of the present study was to evaluate the role of the hypothalamic α7nAChR in the control of energy homeostasis. METHODS: The work was performed on Swiss male mice. Initially, using immunofluorescent staining on brain sections, the presence of α7nAChR in hypothalamic cells regulating energy homeostasis was evaluated. Animals were submitted to stereotaxis in the lateral ventricle and intracerebroventricular stimulation (ICV) was used for the administration of an agonist (PNU) or antagonist (α-bungarotoxin) of α7nAChR. Metabolic parameters were evaluated and the expression of neuropeptides was evaluated in the hypothalamus by real-time PCR and western blot. The expression of hypothalamic neuropeptides was evaluated in mice treated with siRNA or inhibitors of JAK2/STAT3 (AG490 and STATTIC) proteins. We also evaluated food intake in α7nAChR knockout animals (α7KO). Additionally, in mouse hypothalamic cell culture (the mypHoA-POMC/GFP lineage), we evaluated the expression of neuropeptides and pSTAT3 after stimulation with PNU. RESULTS: Our results indicate co-localisation of α7nAChR with α-MSH, AgRP and NPY in hypothalamic cells. Pharmacological activation of α7nAChR reduced food intake and increased hypothalamic POMC expression and decreased NPY and AgRP mRNA levels and the protein content of pAMPK. Inhibition of α7nAChR with an antagonist increased the mRNA content of NPY and AgRP. Inhibition of α7nAChR with siRNA led to the suppression of POMC expression and an increase in AgRP mRNA levels. α7KO mice showed no changes in food intake. Inhibition of proteins involved in the JAK2/STAT3 signalling pathway reversed the effects observed after PNU stimulation. POMC-GFP cells, when treated with PNU, showed increased POMC expression and nuclear translocation of pSTAT3. CONCLUSION: Thus, selective activation of α7nAChR is able to modulate important markers of the response to food intake, suggesting that α7nAChR activation can suppress the expression of orexigenic markers and favour the expression of anorexics using the intracellular JAK2/STAT3 machinery.


Assuntos
Proteína Relacionada com Agouti/metabolismo , Janus Quinase 2/metabolismo , Pró-Opiomelanocortina/metabolismo , Fator de Transcrição STAT3/metabolismo , Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Proteína Relacionada com Agouti/genética , Animais , Benzamidas/farmacologia , Compostos Bicíclicos com Pontes/farmacologia , Bungarotoxinas/farmacologia , Linhagem Celular , Ingestão de Alimentos/efeitos dos fármacos , Metabolismo Energético/efeitos dos fármacos , Janus Quinase 2/antagonistas & inibidores , Janus Quinase 2/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neurônios/metabolismo , Pró-Opiomelanocortina/genética , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Fator de Transcrição STAT3/antagonistas & inibidores , Fator de Transcrição STAT3/genética , Transdução de Sinais/efeitos dos fármacos , Receptor Nicotínico de Acetilcolina alfa7/agonistas , Receptor Nicotínico de Acetilcolina alfa7/antagonistas & inibidores , Receptor Nicotínico de Acetilcolina alfa7/genética
2.
Invest Ophthalmol Vis Sci ; 60(5): 1353-1361, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30934054

RESUMO

Purpose: Motion detection is performed by a unique neural network in the mouse retina. Starburst amacrine cells (SACs), which release acetylcholine and gamma-aminobutyric acid (GABA) into the network, are key neurons in the motion detection pathway. Although GABA contributions to the network have been extensively studied, the role of acetylcholine is minimally understood. Acetylcholine receptors are present in a subset of bipolar, amacrine, and ganglion cells. We focused on α7-nicotinic acetylcholine receptor (α7-nAChR) expression in bipolar cells, and investigated which types of bipolar cells possess α7-nAChRs. Methods: Retinal slice sections were prepared from C57BL/6J and Gus8.4-GFP mice. Specific expression of α7-nAChRs in bipolar cells was examined using α-bungarotoxin (αBgTx)-conjugated Alexa dyes co-labeled with specific bipolar cell markers. Whole-cell recordings were conducted from bipolar cells in retinal slice sections. A selective α7-nAChR agonist, PNU282987, was applied by a puff and responses were recorded. Results: αBgTx fluorescence was observed primarily in bipolar cell somas. We found that α7-nAChRs were expressed by the majority of type 1, 2, 4, and 7 bipolar cells. Whole-cell recordings revealed that type 2 and 7 bipolar cells depolarized by PNU application. In contrast, α7-nAChRs were not detected in most of type 3, 5, 6, and rod bipolar cells. Conclusions: We found that α7-nAChRs are present in bipolar cells in a type-specific manner. Because these bipolar cells provide synaptic inputs to SACs and direction selective ganglion cells, α7-nAChRs may play a role in direction selectivity by modulating these bipolar cells' outputs.


Assuntos
Células Bipolares da Retina/metabolismo , Transmissão Sináptica/fisiologia , Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Animais , Benzamidas/farmacologia , Compostos Bicíclicos com Pontes/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Modelos Animais , Percepção de Movimento/fisiologia , Agonistas Nicotínicos/farmacologia , Receptor Nicotínico de Acetilcolina alfa7/antagonistas & inibidores
3.
Molecules ; 24(8)2019 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-31027274

RESUMO

The chemical composition and biological activity of essential oils isolated from the leaves of Siparuna aspera, Siparuna macrotepala, Piper leticianum, Piper augustum and the rhizome of Hedychium coronarium were evaluated. These species are used medicinally in different ways by the Amazonian communities that live near the Kutukú mountain range. Chemical studies revealed that the main components for the two Siparuna species were germacrene D, bicyclogermacrene, α-pinene, δ-cadinene, δ-elemene, α-copaene and ß-caryophyllene; for the two Piper species ß-caryophyllene, germacrene D, α-(E,E)-farnesene, ß-elemene, bicyclogermacrene, δ-cadinene and for H. coronarium 1,8-cineole, ß-pinene, α-pinene and α-terpineol. The antioxidant activity of all essential oils was evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), photochemiluminescence (PCL) quantitative assays, and DPPH and ABTS bioautographic profiles, with different results for each of them. Antimicrobial activity studies were carried out on three yeasts, six Gram positive and four Gram negative bacteria, by means of the disc diffusion method. The essential oil of H. coronarium showed the most relevant results on L. grayi, K. oxytoca and S. mutans, P. augustum and P. leticianum on S. mutans. An antibacterial bioautographic test for H. coronarium was also carried out and highlighted the potential activity of terpinen-4-ol and 1,8-cineole.


Assuntos
Óleos Voláteis/análise , Zingiberaceae/química , Antibacterianos/análise , Antibacterianos/farmacologia , Compostos Bicíclicos com Pontes/análise , Compostos Bicíclicos com Pontes/farmacologia , Cicloexenos/análise , Cicloexenos/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Monoterpenos/análise , Monoterpenos/farmacologia , Óleos Voláteis/farmacologia , Floresta Úmida , Sesquiterpenos/análise , Sesquiterpenos/farmacologia
4.
Pharmazie ; 74(3): 147-149, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30961679

RESUMO

Mirogabalin, which is a novel ligand for the α2δ subunit of voltage-gated calcium channels, is being developed for treating neuropathic pain including diabetic peripheral neuropathy and postherpetic neuralgia. Mirogabalin possesses unique α2δ subunit binding characteristics and has potent and long-lasting analgesic effects in neuropathic pain models. In the present study, we investigated the effects of mirogabalin on N-type calcium channel currents of the rat dorsal root ganglion (DRG) culture neurons using the whole-cell patch clamp technique. Small or medium DRG neurons were isolated from Sprague-Dawley rats and were incubated for 20 to 24 h with mirogabalin or pregabalin. The DRG neurons were depolarised from a holding potential of -40 mV to +40 mV in steps of 10 mV for 220 ms, and elicited N-type calcium channel currents were recorded. The N-type calcium channel currents were verified by sensitivity to ω-conotoxin GVIA, a selective N-type calcium channel blocker. Mirogabalin inhibited the calcium channel currents of rat DRG neurons at 50 µM, and pregabalin inhibited them at 200 µM. Mirogabalin and pregabalin showed significant differences in the peak current densities at depolarisation to -20 and -10 mV when compared with that shown by the vehicle control. In conclusion, mirogabalin inhibits N-type calcium channel currents in rat DRG culture neurons. The potent and long-lasting analgesic effects of mirogabalin are thought to be associated with its potent and selective binding to α2δ-1 subunits and following functional inhibition of calcium channel currents.


Assuntos
Compostos Bicíclicos com Pontes/farmacologia , Bloqueadores dos Canais de Cálcio/farmacologia , Canais de Cálcio Tipo N/metabolismo , Gânglios Espinais/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Animais , Compostos Bicíclicos com Pontes/química , Bloqueadores dos Canais de Cálcio/química , Células Cultivadas , Conotoxinas/farmacologia , Gânglios Espinais/citologia , Gânglios Espinais/metabolismo , Neurônios/metabolismo , Técnicas de Patch-Clamp , Pregabalina/farmacologia , Ratos , Ratos Sprague-Dawley
5.
Mol Med Rep ; 19(5): 3791-3798, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30864715

RESUMO

The cholinergic anti­inflammatory pathway is considered an attractive approach for the alleviation of inflammatory diseases. Sepsis is characterized by systemic inflammation and widespread organ injury, especially that in the lung. In the present study, we explored the effects of an α7nAChR agonist, PNU­282987, on sepsis­induced lung injury and investigated the mechanisms of PNU­282987 in response to lipopolysaccharide (LPS) stimulation in peritoneal macrophages. Sepsis was induced in C57BL/6 mice via cecal ligation puncture (CLP). Fifty mice were randomly divided into five groups: The sham group treated with vehicle, the sham group treated with PNU­282987, the CLP group treated with vehicle, and the CLP group treated with PNU­282987 (1 mg/kg) 1 h before or 2 h after surgery. All mice were sacrificed at 12 or 24 h after CLP. Both pre­ and post­CLP treatment with PNU­282987 significantly attenuated sepsis­induced lung injury and the release of IL­6 in the bronchoalveolar lavage fluid (BALF). Pre­treatment with PNU­282987 also inhibited sepsis­increased TNF­α and IL­6 production, while post­CLP treatment only inhibited IL­6 production in the lung tissue. Neither pre­ nor post­CLP treatment with PNU­282987 affected IL­6 release in the serum. Furthermore, pretreatment with PNU­282987 resulted in reductions in TNF­α and IL­6 release in a dose­ and time­dependent manner and decreased the phosphorylation levels of p38, JNK and ERK under LPS conditions in peritoneal macrophages. Our results demonstrate that activation of α7nAChR alleviates sepsis­induced lung injury; this effect is associated with the suppression of inflammatory responses via the MAPK pathway, suggesting that α7nAChR is a potential therapeutic target for the treatment of sepsis.


Assuntos
Lesão Pulmonar Aguda/prevenção & controle , Benzamidas/farmacologia , Compostos Bicíclicos com Pontes/farmacologia , Modelos Animais de Doenças , Macrófagos Peritoneais/efeitos dos fármacos , Sepse/complicações , Lesão Pulmonar Aguda/etiologia , Lesão Pulmonar Aguda/patologia , Animais , Células Cultivadas , Lipopolissacarídeos/farmacologia , Macrófagos Peritoneais/metabolismo , Macrófagos Peritoneais/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL
6.
Int J Mol Sci ; 20(6)2019 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-30917511

RESUMO

There is an urgent need to develop novel drugs for osteoporosis which occurs due to estrogen deficiency. Phytoestrogens derived from medicinal plants would be the best alternative to chemical drugs with harmful side effects. The main purpose of the present study was to investigate the effect of ferutinin compared to 17ß-estradiol (E2) on bone mineralization of zebrafish larvae. Regarding the lack of publications, the histology analysis was performed after exposure to E2 to find effective treatment on bone mineralization of developing zebrafish larvae. Then, the larvae were exposed to four concentrations of ferutinin at three time points to assess the mortality, the expression of some related genes and histology of the ceratohyal and hyomandibular of treated larvae. The RT-PCR result of the treatment groups demonstrated the similar expression pattern in the larvae which were exposed to 1.25 µg/mL of ferutinin and 2 µM of E2 at 2 dpf, which confirmed the result of histology analysis. In addition, RT-qPCR of high concentration of ferutinin and E2 demonstrated that bmp2a/b and esr1 were downregulated and upregulated when the larvae were exposed to 5 µg/mL of ferutinin and 10 µM of E2, respectively.


Assuntos
Benzoatos/farmacologia , Calcificação Fisiológica/efeitos dos fármacos , Cicloeptanos/farmacologia , Estradiol/farmacologia , Estrogênios/farmacologia , Sesquiterpenos/farmacologia , Animais , Proteína Morfogenética Óssea 2/genética , Proteína Morfogenética Óssea 2/metabolismo , Compostos Bicíclicos com Pontes/farmacologia , Receptor alfa de Estrogênio/genética , Receptor alfa de Estrogênio/metabolismo , Larva/efeitos dos fármacos , Larva/metabolismo , Peixe-Zebra , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo
7.
Invest Ophthalmol Vis Sci ; 60(2): 570-579, 2019 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-30721274

RESUMO

Purpose: The adult mammalian retina is typically incapable of regeneration when damaged by disease or trauma. Restoration of function would require generation of new adult neurons, something that until recently, mammals were thought to be incapable of doing. However, previous studies from this laboratory have shown that the α7 nicotinic acetylcholine receptor (α7 nAChR) agonist, PNU-282987, induces cell cycle reentry of Müller glia and generation of mature retinal neurons in adult rats, in the absence of detectible injury. This study analyzes how PNU-282987 treatment in RPE leads to robust BrdU incorporation in Müller glia in adult mice and leads to generation of Müller-derived retinal progenitors and neuronal differentiation. Methods: Retinal BrdU incorporation was examined after eye drop application of PNU-282987 in adult wild-type and transgenic mice that contain tamoxifen-inducible tdTomato Müller glia, or after intraocular injection of conditioned medium from PNU-282987-treated cultured RPE cells. Results: PNU-282987 induced robust incorporation of BrdU in all layers of the adult mouse retina. The α7 nAChR agonist was found to stimulate cell cycle reentry of Müller glia and their generation of new retinal progenitors indirectly, via the RPE, in an α7 nAChR-dependent fashion. Conclusions: The results from this study point to RPE as a contributor to Müller glial neurogenic responses. The manipulation of the RPE to stimulate retinal neurogenesis offers a new direction for developing novel and potentially transformative treatments to reverse the loss of neurons associated with neurodegenerative disease, traumatic injury, or aging.


Assuntos
Benzamidas/farmacologia , Compostos Bicíclicos com Pontes/farmacologia , Células Ependimogliais/fisiologia , Neurogênese/fisiologia , Neuroglia/fisiologia , Agonistas Nicotínicos/farmacologia , Epitélio Pigmentado da Retina/efeitos dos fármacos , Receptor Nicotínico de Acetilcolina alfa7/agonistas , Animais , Bromodesoxiuridina/metabolismo , Ciclo Celular/fisiologia , Células Cultivadas , Células Ependimogliais/citologia , Feminino , Masculino , Camundongos , Camundongos Transgênicos , Fator de Transcrição PAX6/metabolismo , Ratos , Epitélio Pigmentado da Retina/citologia , Epitélio Pigmentado da Retina/metabolismo , Células-Tronco/fisiologia
8.
Vet Parasitol ; 266: 84-87, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30736953

RESUMO

Cystic echinococcosis is of great public health importance. Surgery is the efficient treatment for this infection. To minimize the risk of cyst intraoperative leakage, using scolicidals is crucial. To date, any efficacious scolicidal without side effect has not been introduced. Since essential oils of Pelargonium roseum and Ferula gummosa have shown several bioactivities, we evaluated their potential against protoscoleces of E. granulosus using albendazole as positive control. Furthermore, chemical composition of both essential oils was analyzed by gas chromatography-mass spectrometry (GC-MS) analyses, and their main constituents were also evaluated for scolicidal activity. Different concentrations of essential oils and their two main constituents were tested for scolicidal activity. Mortality rate was measured by eosin staining. Results of GC-MS revealed citronellol and ß-pinene as the main constituents of P. roseum and F. gummosa essential oils, respectively. After 60 min of exposure to 50 µg/mL of P. roseum and F. gummosa, mean mortality rate of protoscoleces was 100%. However, ß-pinene and citronellol at the same time point with only 10 µg/mL concentrations resulted in approximately higher than 80% mortality. Essential oils of P. roseum and F. gummosa showed significant toxic effect on E. granulosus with 50% lethal concentration (LC50) values of 8.52 and 17.18 µg/mL, respectively. Based on the LC50 values, ß-pinene (2.20 µg/mL) was the most potent scolicidal agent in the present study. The overall toxicity of ß-pinene and citronellol was significantly higher than the whole essential oils of F. gummosa and P. roseum. Based on these results, ß-pinene and citronellol can be considered as candidate ingredients for the development of green scolicidals.


Assuntos
Echinococcus granulosus/efeitos dos fármacos , Ferula/química , Óleos Voláteis/farmacologia , Pelargonium/química , Óleos Vegetais/farmacologia , Albendazol/farmacologia , Animais , Compostos Bicíclicos com Pontes/farmacologia , Cromatografia Gasosa-Espectrometria de Massas , Monoterpenos/farmacologia
9.
Molecules ; 24(2)2019 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-30634693

RESUMO

Here, we investigated the anti-oxidant and anti-melanogenic effects of pomelo peel essential oil (PPEO) from pomelo cv. Guan Xi. The volatile chemical composition of PPEO was analyzed with gas chromatography⁻mass spectrometry (GC/MS). The most abundant component of PPEO was limonene (55.92%), followed by ß-myrcene (31.17%), and ß-pinene (3.16%). PPEO showed strong anti-oxidant activities against 1,1-diphenyl-2-picryhydrazyl (DPPH) (,2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) (, and superoxide anion free radicals. Based on the B16 melanoma cell system, the effects of PPEO on the viability and morphology of B16 cells and the production of melanin were evaluated. The results revealed that PPEO at concentrations below 50 µg/mL could decrease the melanin content without affecting cell viability and morphology. Intracellular tyrosinase (TYR) activity and Western blot analysis showed that PPEO could down-regulate the expression level of TYR in B16 cells and dose-dependently inhibit TYR activity (by a maximum of 64.54%). Meanwhile, the main active components responsible for the effect are citral and ß-myrcene. In conclusion, PPEO has good anti-oxidant and anti-melanogenic activity, and thus can be widely used as a natural antioxidant in the food, pharmaceutical, and cosmetic industries.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Antioxidantes/farmacologia , Citrus/química , Melanoma/enzimologia , Óleos Voláteis/farmacologia , Animais , Antineoplásicos Fitogênicos/química , Antioxidantes/química , Compostos Bicíclicos com Pontes/química , Compostos Bicíclicos com Pontes/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Regulação para Baixo , Cromatografia Gasosa-Espectrometria de Massas , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Limoneno/química , Limoneno/farmacologia , Melanoma/tratamento farmacológico , Camundongos , Monofenol Mono-Oxigenase/metabolismo , Monoterpenos/química , Monoterpenos/farmacologia , Óleos Voláteis/química , Óleos Vegetais/química , Óleos Vegetais/farmacologia
10.
Methods Cell Biol ; 149: 179-194, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30616818

RESUMO

The superfamily of G protein-coupled receptors (GPCRs) represents the largest group of cell surface receptors in the human body. It is estimated that around 40% of the drugs currently on the market target GPCRs. As only a very small number of GPCRs is targeted by these marketed drugs, the potential of GPCRs as novel drug targets remains enormous. As opposed to conventional in vitro assays, label-free cellular assays using a biosensor provide new opportunities for studying GPCRs. Integrated receptor-mediated responses are measured in real-time rather than a single downstream signaling pathway, without the need for the use of any label (e.g., fluorescent or radioactive). Here, we describe a protocol to study GPCR pharmacology using the label-free whole cell impedance-based biosensor system xCELLigence. This assay allows quantification of compound-induced GPCR-mediated responses in real-time. Finally, we have also discussed the analysis and interpretation of the results obtained with this assay using the mGlu2 receptor as a model system.


Assuntos
Bioensaio/métodos , Impedância Elétrica , Receptores Acoplados a Proteínas-G/metabolismo , Coloração e Rotulagem , Aminoácidos/farmacologia , Animais , Técnicas Biossensoriais , Compostos Bicíclicos com Pontes/farmacologia , Células CHO , Contagem de Células , Cricetinae , Cricetulus , Humanos , Ligantes , Receptores Acoplados a Proteínas-G/agonistas , Receptores de Glutamato Metabotrópico/metabolismo , Transdução de Sinais/efeitos dos fármacos , Xantenos/farmacologia
11.
Bioorg Med Chem ; 27(2): 338-342, 2019 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-30545734

RESUMO

Isocarbacyclin is a valuable synthetic analogue of prostacyclin with potential neuroprotective effects for the treatment of ischemic stroke. Herein, we describe the synthesis of isocarbacyclin and bicyclic analogues in only 7-10 steps, with the ω-side chain diversified at a late stage. A combination of new reaction design, function-oriented synthesis, and late-stage diversification led to a series of compounds that were tested for their neuroprotective activities. Efforts toward the synthesis of tricyclic analogues of isocarbacyclin, using the same combination of metal-catalyzed reactions, is also described.


Assuntos
Compostos Bicíclicos com Pontes/farmacologia , Epoprostenol/análogos & derivados , Fármacos Neuroprotetores/farmacologia , Animais , Isquemia Encefálica/tratamento farmacológico , Compostos Bicíclicos com Pontes/síntese química , Compostos Bicíclicos com Pontes/química , Epoprostenol/síntese química , Epoprostenol/química , Epoprostenol/farmacologia , Camundongos , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/síntese química , Fármacos Neuroprotetores/química , Estereoisomerismo , Acidente Vascular Cerebral/tratamento farmacológico
12.
Biomed Pharmacother ; 111: 359-366, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30594048

RESUMO

The cholinergic anti-inflammatory pathway modulates cytokine release by activating alpha-7 nicotinic acetylcholine receptors (α7nAChR) in monocytes/macrophages. We aimed to determine the role of α7nAChR in lupus nephritis (LN). We enrolled 36 inactive and 35 active LN patients, 34 primary glomerulonephritis patients, and 35 healthy controls. Peripheral blood monocytes were isolated, and mRNA expression of α7nAChR, interleukin (IL)-1ß, IL-6, IL-10, and tumor necrosis factor-alpha (TNF-α) in monocytes was measured. α7nAChR and IL-10 mRNA levels were significantly decreased, but IL-6 was increased, in LN patients compared with healthy controls or glomerulonephritis patients (all P < 0.01). Interestingly, α7nAChR mRNA levels were negatively correlated to SLEDAI (r = -0.68, P < 0.01), anti-dsDNA (r = -0.38, P < 0.05), and proteinuria (r = -0.49, P < 0.01) levels, and positively correlated to serum complement C3 levels (r = 0.38, P < 0.05) in patients with active LN. Furthermore, α7nAChR mRNA levels were negatively correlated to TNF-α (r = -0.50, P < 0.01), IL-1ß (r = -0.42, P < 0.05), IL-6 (r = -0.69, P < 0.01) mRNA levels, and positively correlated to IL-10 (r = 0.45, P < 0.01). TNF-α, IL-1ß, and IL-6 protein levels in the supernatant of cultured monocytes from active LN patients were significantly higher, while IL-10 was lower, than that of healthy controls. PNU-282987, an α7nAChR agonist, significantly decreased TNF-α, IL-1ß, and IL-6 but increased IL-10 in the monocyte culture supernatant of active LN patients, which were abolished by an α7nAChR antagonist methyllycaconitine. The effects of PNU-282987 were confirmed in lipopolysaccharides-stimulated monocytes. Taken together, these findings suggest that decrease in α7nAChR mRNA levels may play a role in LN and that activation of α7nAChR may inhibit inflammation in LN.


Assuntos
Citocinas/metabolismo , Mediadores da Inflamação/metabolismo , Leucócitos Mononucleares/metabolismo , Nefrite Lúpica/metabolismo , RNA Mensageiro/metabolismo , Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Adulto , Benzamidas/farmacologia , Compostos Bicíclicos com Pontes/farmacologia , Células Cultivadas , Citocinas/antagonistas & inibidores , Feminino , Humanos , Mediadores da Inflamação/antagonistas & inibidores , Leucócitos Mononucleares/efeitos dos fármacos , Nefrite Lúpica/patologia , Masculino , Pessoa de Meia-Idade , Agonistas Nicotínicos/farmacologia , Receptor Nicotínico de Acetilcolina alfa7/agonistas
13.
Chem Commun (Camb) ; 54(56): 7810-7813, 2018 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-29946609

RESUMO

Here we report the 18F labeling of a prostate specific membrane antigen (PSMA) ligand via a strain promoted oxa-dibenzocyclooctyne (ODIBO)- or bicyclo[6.1.0]nonyne (BCN)-azide reaction. Although ODIBO reacts with azide 20 fold faster than BCN, in vivo PET imaging suggests that 18F-BCN-azide-PSMA demonstrated much higher tumor uptake and a much higher tumor to background contrast.


Assuntos
Alquinos/química , Antígenos de Superfície/metabolismo , Azidas/química , Compostos Bicíclicos com Pontes/metabolismo , Ciclo-Octanos/metabolismo , Glutamato Carboxipeptidase II/metabolismo , Hidrocarbonetos Fluorados/metabolismo , Compostos Radiofarmacêuticos/metabolismo , Animais , Compostos Bicíclicos com Pontes/síntese química , Compostos Bicíclicos com Pontes/química , Compostos Bicíclicos com Pontes/farmacologia , Ciclo-Octanos/síntese química , Ciclo-Octanos/química , Ciclo-Octanos/farmacologia , Dipeptídeos/metabolismo , Radioisótopos de Flúor , Compostos Heterocíclicos com 1 Anel/metabolismo , Humanos , Hidrocarbonetos Fluorados/síntese química , Hidrocarbonetos Fluorados/química , Hidrocarbonetos Fluorados/farmacologia , Interações Hidrofóbicas e Hidrofílicas , Cinética , Ligantes , Masculino , Camundongos , Tomografia por Emissão de Pósitrons , Neoplasias da Próstata/diagnóstico por imagem , Ligação Proteica , Compostos Radiofarmacêuticos/síntese química , Compostos Radiofarmacêuticos/química , Compostos Radiofarmacêuticos/farmacologia , Ensaios Antitumorais Modelo de Xenoenxerto
14.
Biomed Pharmacother ; 105: 267-273, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29860218

RESUMO

Estrogen is considered a risk factor for breast cancer since it promotes breast-cell proliferation. The jaesckeanadiol-3-p-hydroxyphenylpropanoate, a hemi-synthetic analogue of the natural phytoestrogen ferutinin (jaesckeanadiol-p-hydroxybenzoate), is designed to be devoid of estrogenic activity. This analogue induces a cytotoxic effect 30 times higher than that of ferutinin towards MCF-7 breast cancer cell line. We compared these two compounds with respect to their effect on proliferation, cell cycle distribution and cancer stem-like cells in the MCF-7 cell line. Treatment with ferutinin (30 µM) and its analogue (1 µM) produced significant accumulation of cells at the pre G0/G1 cell cycle phase and triggered apoptosis. Importantly, this compound retains its anti-proliferative activity against breast cancer stem/progenitor cells that are naturally insensitive to ferutinin at the same dose. These results position ferutinin analogue as an effective compound inhibiting the proliferation of estrogen-dependent breast cancer cells and consistently targeting their stem-like cells.


Assuntos
Benzoatos/química , Benzoatos/farmacologia , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Cicloeptanos/química , Cicloeptanos/farmacologia , Receptores Estrogênicos/metabolismo , Sesquiterpenos/química , Sesquiterpenos/farmacologia , Apoptose/efeitos dos fármacos , Compostos Bicíclicos com Pontes/química , Compostos Bicíclicos com Pontes/farmacologia , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Autorrenovação Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Feminino , Humanos , Células MCF-7 , Células-Tronco Neoplásicas/efeitos dos fármacos , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia , Esferoides Celulares/efeitos dos fármacos , Esferoides Celulares/metabolismo , Esferoides Celulares/patologia
15.
Bioorg Med Chem ; 26(12): 3345-3351, 2018 07 23.
Artigo em Inglês | MEDLINE | ID: mdl-29748146

RESUMO

Bicyclo[3,3,1]non-2-ene was used as a novel three-dimensional skeleton for the design and synthesis of hypoxia-inducible factor (HIF)-1 inhibitors. Among the compounds synthesized, compound 4b was found to be a potent inhibitor of HIF-1α protein accumulation under hypoxia and inhibited HIF-1α transcriptional activity in HeLa (human cervical carcinoma) cells (half maximal inhibitory concentration [IC50] = 3.0 µM). The inhibition of HIF-1α accumulation induced by compound 4b was attenuated by treating the cells with MG132, a proteasome inhibitor, in a concentration-dependent manner, indicating that the compound 4b induces oxygen-independent proteasomal degradation of HIF-1α.


Assuntos
Compostos Bicíclicos com Pontes/química , Subunidade alfa do Fator 1 Induzível por Hipóxia/antagonistas & inibidores , Compostos Bicíclicos com Pontes/síntese química , Compostos Bicíclicos com Pontes/farmacologia , Desenho de Drogas , Células HeLa , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Leupeptinas/farmacologia , Complexo de Endopeptidases do Proteassoma/química , Complexo de Endopeptidases do Proteassoma/metabolismo , Transcrição Genética/efeitos dos fármacos
16.
Eur J Med Chem ; 152: 401-416, 2018 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-29751234

RESUMO

The cholinergic pathways in the central nervous system (CNS) of animals and humans are important for cognitive and behavioural functions. Until a few years ago, it was thought that the key molecules transducing the cholinergic message were the metabotropic muscarinic receptors, but it is now known that ionotropic neuronal nicotinic receptors (nAChRs) are also involved. Based on recent studies, we prepared a small library of novel 3-substituted-3,6-diazabicyclo [3.1.1]heptanes, whose binding activity and functionality have been assayed. Among the synthesized compounds, the 3-(anilino)pyridine series resulted in the most interesting compounds with α4ß2Ki values ranging from 0.0225 nM (12g) to 2.06 nM (12o).


Assuntos
Compostos de Anilina/farmacologia , Compostos Bicíclicos com Pontes/farmacologia , Receptores Nicotínicos/metabolismo , Compostos de Anilina/química , Animais , Compostos Bicíclicos com Pontes/química , Linhagem Celular , Relação Dose-Resposta a Droga , Humanos , Ligantes , Estrutura Molecular , Ratos , Relação Estrutura-Atividade
17.
Acta Trop ; 185: 98-106, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29709631

RESUMO

Several studies have shown that odors of plant and animal origin can be developed into lures for use in surveillance of mosquito vectors of infectious diseases. However, the effect of combining plant- and mammalian-derived odors into an improved lure for monitoring both nectar- and blood-seeking mosquito populations in traps is yet to be explored. Here we used both laboratory dual choice olfactometer and field assays to investigate responses of the malaria vector, Anopheles gambiae, to plant- and mammalian-derived compounds and a combined blend derived from these two odor sources. Using subtractive bioassays in dual choice olfactometer we show that a 3-component terpenoid plant-derived blend comprising (E)-linalool oxide, ß-pinene, ß-ocimene was more attractive to females of An. gambiae than (E)-linalool oxide only (previously found attractive in field trials) and addition of limonene to this blend antagonized its attractiveness. Likewise, a mammalian-derived lure comprising the aldehydes heptanal, octanal, nonanal and decanal, was more preferred than (E)-linalool oxide. Surprisingly, combining the plant-derived 3-component blend with the mammalian derived 4-component blend attracted fewer females of An. gambiae than the individual blends in laboratory assays. However, this pattern was not replicated in field trials, where we observed a dose-dependent effect on trap catches while combining both blends with significantly improved trap catches at higher doses. The observed dose-dependent attractiveness for An. gambiae has practical implication in the design of vector control strategies involving kairomones from plant- and mammalian-based sources.


Assuntos
Anopheles/fisiologia , Mamíferos , Mosquitos Vetores/fisiologia , Odorantes , Feromônios/farmacologia , Plantas , Aldeídos/farmacologia , Alcenos/farmacologia , Animais , Anopheles/efeitos dos fármacos , Compostos Bicíclicos com Pontes/farmacologia , Cicloexanóis/farmacologia , Cicloexenos/farmacologia , Feminino , Limoneno , Malária/transmissão , Monoterpenos/farmacologia , Controle de Mosquitos , Mosquitos Vetores/efeitos dos fármacos , Terpenos/farmacologia , Compostos de Tritil/farmacologia
18.
Bioorg Med Chem Lett ; 28(6): 1043-1049, 2018 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-29486970

RESUMO

A series of isoquinuclidine benzamides as glycine uptake inhibitors for the treatment of schizophrenia are described. Potency, lipophilicity, and intrinsic human microsomal clearance were parameters for optimization. Potency correlated with the nature of the ortho substituents of the benzamide ring, and reductions in lipophilicity could be achieved through heteroatom incorporation in the benzamide and pendant phenyl moieties. Improvements in human CLint were achieved through changes in ring size and the N-alkyl group of the isoquinuclidine itself, with des-alkyl derivatives (40-41, 44) demonstrating the most robust microsomal stability. Dimethylbenzamide 9 was tested in a mouse MK801 LMA assay and had a statistically significant attenuation of locomotor activity at 3 and 10 µmol/kg compared to control.


Assuntos
Benzamidas/farmacologia , Compostos Bicíclicos com Pontes/farmacologia , Proteínas da Membrana Plasmática de Transporte de Glicina/antagonistas & inibidores , Administração Oral , Animais , Benzamidas/administração & dosagem , Benzamidas/química , Compostos Bicíclicos com Pontes/administração & dosagem , Compostos Bicíclicos com Pontes/química , Relação Dose-Resposta a Droga , Humanos , Injeções Intravenosas , Locomoção/efeitos dos fármacos , Masculino , Camundongos , Estrutura Molecular , Ratos , Ratos Sprague-Dawley , Relação Estrutura-Atividade
19.
Bioorg Med Chem ; 26(5): 1026-1034, 2018 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-29422332

RESUMO

The TGFß-TGFßR signaling pathway has been reported to play a protective role in the later stages of tumorigenesis via increasing immunosuppressive Treg cells and facilitating the epithelial to mesenchymal transition (EMT). Therefore, inhibition of TGFßR has the potential to enhance antitumor immunity. Herein we disclose the identification and optimization of novel heterobicyclic inhibitors of TGFßRI that demonstrate potent inhibition of SMAD phosphorylation. Application of structure-based drug design to the novel pyrrolotriazine chemotype resulted in improved binding affinity (Ki apparent = 0.14 nM), long residence time (T1/2 > 120 min) and significantly improved potency in the PSMAD cellular assay (IC50 = 24 nM). Several analogs inhibited phosphorylation of SMAD both in vitro and in vivo. Additionally, inhibition of TGFß-stimulated phospho-SMAD was observed in primary human T cells.


Assuntos
Compostos Bicíclicos com Pontes/química , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Receptores de Fatores de Crescimento Transformadores beta/antagonistas & inibidores , Sítios de Ligação , Compostos Bicíclicos com Pontes/síntese química , Compostos Bicíclicos com Pontes/farmacologia , Células Cultivadas , Cristalografia por Raios X , Desenho de Drogas , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Humanos , Simulação de Dinâmica Molecular , Fosforilação , Ligação Proteica , Estrutura Terciária de Proteína , Proteínas Serina-Treonina Quinases/metabolismo , Pirróis/síntese química , Pirróis/química , Pirróis/metabolismo , Receptor do Fator de Crescimento Transformador beta Tipo I , Receptores de Fatores de Crescimento Transformadores beta/metabolismo , Proteínas Smad/metabolismo , Relação Estrutura-Atividade , Linfócitos T/citologia , Linfócitos T/metabolismo , Tiazinas/síntese química , Tiazinas/química , Tiazinas/metabolismo
20.
Bioorg Med Chem Lett ; 28(6): 1111-1115, 2018 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-29426770

RESUMO

The diastereoselective synthesis and structure activity relationship (SAR) of a series of fused cyclopropyl-3-amino-2,4-oxazine (2-oxa-4-azabicyclo[4.1.0]hept-3-en-3-amine)-containing BACE inhibitors is described. Through these efforts compound 2 was identified as a potent (cell IC50 = 15 nM) BACE inhibitor with acceptable ADME properties. When tested in vivo, compound 2 demonstrated a significant reduction of brain and cerebral spinal fluid (CSF) Aß40 levels (46% and 66%, respectively) in a rat pharmacodynamic study and thus represents a suitable starting point for the further development of in vivo efficacious compounds for the treatment of Alzheimer's disease.


Assuntos
Secretases da Proteína Precursora do Amiloide/antagonistas & inibidores , Ácido Aspártico Endopeptidases/antagonistas & inibidores , Compostos Aza/farmacologia , Compostos Bicíclicos com Pontes/farmacologia , Inibidores Enzimáticos/farmacologia , Secretases da Proteína Precursora do Amiloide/metabolismo , Animais , Ácido Aspártico Endopeptidases/metabolismo , Compostos Aza/síntese química , Compostos Aza/química , Compostos Bicíclicos com Pontes/síntese química , Compostos Bicíclicos com Pontes/química , Cristalografia por Raios X , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Humanos , Modelos Moleculares , Estrutura Molecular , Ratos , Ratos Sprague-Dawley , Estereoisomerismo , Relação Estrutura-Atividade
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