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[PMID]: 28699792 [Au] Autor: Zhou Y; Cai M [Ad] Endereço: a Department of Chemistry and Biochemistry , University of Arizona , Tucson , AZ USA. [Ti] Título: Novel approaches to the design of bioavailable melanotropins. [So] Source: Expert Opin Drug Discov;12(10):1023-1030, 2017 Oct. [Is] ISSN: 1746-045X [Cp] País de publicação: England [La] Idioma: eng [Ab] Resumo: INTRODUCTION: The melanocortin system is a primordial and critical system for survival, involved in a wide variety of physiological functions. It includes melanocortin receptors (MCRs) and melanotropin ligands (MCLs). MCRs are important drug targets that can regulate several key physiological processes. Extensive efforts have been made to develop peptide and peptidomimetics targeting melanocortin receptors including MC1R, MC3R, MC4R and MC5R. Most research is focused on developing potent and selective melanotropins. However, developing bioavailable melanotropins remains challenging. Areas covered: Herein, the authors summarize promising strategies for developing bioavailable MCLs by using cyclized N-methylated melanotropins, and using cyclotide and tetrapeptide as templates. They discuss their unique advantages in oral availability and targeting MCRs in the central nervous system or in peripheral tissues. Finally, they discuss the observed differences in thepharmacology of MCRs between in vitro and in vivo tests. Expert opinion: N-methylated cyclized melanotropins have great potential to become bio- available drugs targeting MCRs in the brain, while MCR-grafted cyclotides tend to target MCRs in peripheral tissue. A better understanding of the biased signaling process is a new challenge and opportunity for the future discovery of bioavailable MCLs. [Mh] Termos MeSH primário: Desenho de Drogas Hormônios Estimuladores de Melanócitos/síntese química Receptores de Melanocortina/metabolismo [Mh] Termos MeSH secundário: Animais Disponibilidade Biológica Ciclotídeos/síntese química Ciclotídeos/farmacocinética Ciclotídeos/farmacologia Seres Humanos Ligantes Hormônios Estimuladores de Melanócitos/química Hormônios Estimuladores de Melanócitos/farmacocinética Peptídeos/síntese química Peptídeos/farmacocinética Peptídeos/farmacologia Peptidomiméticos/síntese química Peptidomiméticos/farmacocinética Peptidomiméticos/farmacologia Distribuição Tecidual [Pt] Tipo de publicação: JOURNAL ARTICLE; REVIEW [Nm] Nome de substância: 0 (Cyclotides); 0 (Ligands); 0 (Peptides); 0 (Peptidomimetics); 0 (Receptors, Melanocortin); 9002-79-3 (Melanocyte-Stimulating Hormones) [Em] Mês de entrada: 1709 [Cu] Atualização por classe: 170922 [Lr] Data última revisão: 170922 [Sb] Subgrupo de revista: IM [Da] Data de entrada para processamento: 170713 [St] Status: MEDLINE [do] DOI: 10.1080/17460441.2017.1351940

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[PMID]: 28621949 [Au] Autor: Narayani M; Chadha A; Srivastava S [Ad] Endereço: Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras , Chennai 600036, India. [Ti] Título: Cyclotides from the Indian Medicinal Plant Viola odorata (Banafsha): Identification and Characterization. [So] Source: J Nat Prod;80(7):1972-1980, 2017 Jul 28. [Is] ISSN: 1520-6025 [Cp] País de publicação: United States [La] Idioma: eng [Ab] Resumo: Cyclotides are cyclic cystine knotted macrocyclic plant peptides that have several promising applications. This study was undertaken to detect and identify known and new cyclotides in Viola odorata, a commercially important medicinal plant, from three geographical locations in India. The number of cyclotides in the plant varied with the tissue (leaves, petioles, flowers, runners, and roots) and with geographical locations in India. Using liquid chromatography coupled to Fourier transform mass spectrometry (FTMS), 166 cyclotide-like masses were observed to display cyclotide-diagnostic mass shifts following reduction, alkylation, and digestion, and 71 of these were positively identified based on automated spectrum matching. Of the remaining 95 putative cyclotides observed, de novo peptide sequencing of three new cyclotides, namely, vodo I1 (1), vodo I2 (2), and vodo I3 (3), was carried out with tandem mass spectrometry. [Mh] Termos MeSH primário: Ciclotídeos/isolamento & purificação Plantas Medicinais/química Viola/química [Mh] Termos MeSH secundário: Sequência de Aminoácidos Cromatografia Líquida de Alta Pressão Ciclotídeos/química Índia Estrutura Molecular Ressonância Magnética Nuclear Biomolecular Folhas de Planta/química [Pt] Tipo de publicação: JOURNAL ARTICLE [Nm] Nome de substância: 0 (Cyclotides); 0 (vodo I1); 0 (vodo I2); 0 (vodo I3) [Em] Mês de entrada: 1709 [Cu] Atualização por classe: 170915 [Lr] Data última revisão: 170915 [Sb] Subgrupo de revista: IM [Da] Data de entrada para processamento: 170617 [St] Status: MEDLINE [do] DOI: 10.1021/acs.jnatprod.6b01004

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[PMID]: 28544675 [Au] Autor: Gould A; Camarero JA [Ad] Endereço: Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA, 90089-9121, USA. [Ti] Título: Cyclotides: Overview and Biotechnological Applications. [So] Source: Chembiochem;18(14):1350-1363, 2017 Jul 18. [Is] ISSN: 1439-7633 [Cp] País de publicação: Germany [La] Idioma: eng [Ab] Resumo: Cyclotides are globular microproteins with a unique head-to-tail cyclized backbone, stabilized by three disulfide bonds forming a cystine knot. This unique circular backbone topology and knotted arrangement of three disulfide bonds makes them exceptionally stable to chemical, thermal, and biological degradation compared to other peptides of similar size. In addition, cyclotides have been shown to be highly tolerant to sequence variability, aside from the conserved residues forming the cystine knot. Cyclotides can also cross cellular membranes and are able to modulate intracellular protein-protein interactions, both in vitro and in vivo. All of these features make cyclotides highly promising as leads or frameworks for the design of peptide-based diagnostic and therapeutic tools. This article provides an overview on cyclotides and their applications as molecular imaging agents and peptide-based therapeutics. [Mh] Termos MeSH primário: Biotecnologia/métodos Ciclotídeos/química Ciclotídeos/uso terapêutico Imagem Molecular/métodos [Mh] Termos MeSH secundário: Animais Ciclotídeos/farmacologia Seres Humanos Modelos Moleculares Sondas Moleculares/química Sondas Moleculares/farmacologia Sondas Moleculares/uso terapêutico Plantas/química Proteínas/antagonistas & inibidores Proteínas/metabolismo [Pt] Tipo de publicação: JOURNAL ARTICLE; REVIEW [Nm] Nome de substância: 0 (Cyclotides); 0 (Molecular Probes); 0 (Proteins) [Em] Mês de entrada: 1710 [Cu] Atualização por classe: 171010 [Lr] Data última revisão: 171010 [Sb] Subgrupo de revista: IM [Da] Data de entrada para processamento: 170526 [St] Status: MEDLINE [do] DOI: 10.1002/cbic.201700153

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[PMID]: 28256223 [Au] Autor: Grage SL; Sani MA; Cheneval O; Henriques ST; Schalck C; Heinzmann R; Mylne JS; Mykhailiuk PK; Afonin S; Komarov IV; Separovic F; Craik DJ; Ulrich AS [Ad] Endereço: Institute of Biological Interfaces (IBG-2), Karlsruhe Institute of Technology, Karlsruhe, Germany. [Ti] Título: Orientation and Location of the Cyclotide Kalata B1 in Lipid Bilayers Revealed by Solid-State NMR. [So] Source: Biophys J;112(4):630-642, 2017 Feb 28. [Is] ISSN: 1542-0086 [Cp] País de publicação: United States [La] Idioma: eng [Ab] Resumo: Cyclotides are ultra-stable cyclic disulfide-rich peptides from plants. Their biophysical effects and medically interesting activities are related to their membrane-binding properties, with particularly high affinity for phosphatidylethanolamine lipids. In this study we were interested in understanding the molecular details of cyclotide-membrane interactions, specifically with regard to the spatial orientation of the cyclotide kalata B1 from Oldenlandia affinis when embedded in a lipid bilayer. Our experimental approach was based on the use of solid-state F-NMR of oriented bilayers in conjunction with the conformationally restricted amino acid L-3-(trifluoromethyl)bicyclopent-[1.1.1]-1-ylglycine as an orientation-sensitive F-NMR probe. Its rigid connection to the kalata B1 backbone scaffold, together with the well-defined structure of the cyclotide, allowed us to calculate the protein alignment in the membrane directly from the orientation-sensitive F-NMR signal. The hydrophobic and polar residues on the surface of kalata B1 form well-separated patches, endowing this cyclotide with a pronounced amphipathicity. The peptide orientation, as determined by NMR, showed that this amphipathic structure matches the polar/apolar interface of the lipid bilayer very well. A location in the amphiphilic headgroup region of the bilayer was supported by N-NMR of uniformly labeled protein, and confirmed using solid-state P- and H-NMR. P-NMR relaxation data indicated a change in lipid headgroup dynamics induced by kalata B1. Changes in the H-NMR order parameter profile of the acyl chains suggest membrane thinning, as typically observed for amphiphilic peptides embedded near the polar/apolar bilayer interface. Furthermore, from the F-NMR analysis two important charged residues, E7 and R28, were found to be positioned equatorially. The observed location thus would be favorable for the postulated binding of E7 to phosphatidylethanolamine lipid headgroups. Furthermore, it may be speculated that this pair of side chains could promote oligomerization of kalata B1 through electrostatic intermolecular contacts via their complementary charges. [Mh] Termos MeSH primário: Ciclotídeos/química Ciclotídeos/metabolismo Bicamadas Lipídicas/metabolismo [Mh] Termos MeSH secundário: Sequência de Aminoácidos Espectroscopia de Ressonância Magnética Modelos Moleculares Oldenlandia/metabolismo Conformação Proteica [Pt] Tipo de publicação: JOURNAL ARTICLE [Nm] Nome de substância: 0 (Cyclotides); 0 (Lipid Bilayers); 0 (kalata B1) [Em] Mês de entrada: 1707 [Cu] Atualização por classe: 170713 [Lr] Data última revisão: 170713 [Sb] Subgrupo de revista: IM [Da] Data de entrada para processamento: 170304 [St] Status: MEDLINE

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[PMID]: 28249194 [Au] Autor: Craik DJ; Du J [Ad] Endereço: Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland 4072, Australia. Electronic address: d.craik@imb.uq.edu.au. [Ti] Título: Cyclotides as drug design scaffolds. [So] Source: Curr Opin Chem Biol;38:8-16, 2017 Jun. [Is] ISSN: 1879-0402 [Cp] País de publicação: England [La] Idioma: eng [Ab] Resumo: Cyclotides are ultra-stable peptides derived from plants. They are around 30 amino acids in size and are characterized by their head-to-tail cyclic backbone and cystine knot. Their exceptional stability and tolerance to sequence substitutions has led to their use as frameworks in drug design. This article describes recent developments in this field, particularly developments over the last two years relating to the grafting of bioactive peptide sequences into the cyclic cystine knot framework of cyclotides to stabilize the sequences. Grafted cyclotides have now been developed that interact with protein or enzyme targets, both extracellular and intracellular, as well as with cell surface receptors and membranes. [Mh] Termos MeSH primário: Ciclotídeos/química Desenho de Drogas [Mh] Termos MeSH secundário: Sequência de Aminoácidos Animais Ciclotídeos/síntese química Ciclotídeos/farmacologia Seres Humanos [Pt] Tipo de publicação: JOURNAL ARTICLE; REVIEW [Nm] Nome de substância: 0 (Cyclotides) [Em] Mês de entrada: 1707 [Cu] Atualização por classe: 170719 [Lr] Data última revisão: 170719 [Sb] Subgrupo de revista: IM [Da] Data de entrada para processamento: 170302 [St] Status: MEDLINE

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[PMID]: 28085267 [Au] Autor: Troeira Henriques S; Craik DJ [Ad] Endereço: Institute for Molecular Bioscience, The University of Queensland , Brisbane, 4072 QLD, Australia. [Ti] Título: Cyclotide Structure and Function: The Role of Membrane Binding and Permeation. [So] Source: Biochemistry;56(5):669-682, 2017 Feb 07. [Is] ISSN: 1520-4995 [Cp] País de publicação: United States [La] Idioma: eng [Ab] Resumo: There is growing interest in the use of peptides as therapeutic drugs and, in particular, in the potential of cyclotides, a family of cyclic peptides with remarkable stability and amenability to sequence engineering, as scaffolds in drug design. As well as having an ultrastable structure, many natural cyclotides have intrinsic biological activities with potential pharmaceutical or agricultural applications. Some cyclotides also have the ability to cross membrane barriers and to enter into cells; in particular, cyclotides that belong to the Möbius and bracelet subfamilies have been found to harbor lipid-binding domains, which allow for the specific recognition of phosphatidylethanolamine phospholipids in biological membranes. This lipid selectivity is intimately correlated with the highly conserved three-dimensional structures of cyclotides and is important for their reported biological properties and cell penetration ability. The membrane binding features of Möbius and bracelet cyclotides contrast with the lack of membrane binding of trypsin inhibitor cyclotides, which have physicochemical properties and bioactivities different from those of the other two subfamilies of cyclotides but are also able to enter cells. This review discusses the structures of cyclotides with regard to their myriad of biological activities and describes the role of membrane binding in their functions and ability to enter cells. [Mh] Termos MeSH primário: Antibacterianos/química Ciclotídeos/química Inseticidas/química Fosfatidiletanolaminas/química Inibidores da Tripsina/química [Mh] Termos MeSH secundário: Sequência de Aminoácidos Animais Antibacterianos/farmacologia Permeabilidade da Membrana Celular/efeitos dos fármacos Ciclotídeos/farmacologia Dípteros/efeitos dos fármacos Dípteros/fisiologia Bactérias Gram-Negativas/efeitos dos fármacos Bactérias Gram-Negativas/crescimento & desenvolvimento Bactérias Gram-Positivas/efeitos dos fármacos Bactérias Gram-Positivas/crescimento & desenvolvimento Seres Humanos Inseticidas/farmacologia Lepidópteros/efeitos dos fármacos Lepidópteros/fisiologia Ligação Proteica Estabilidade Proteica Alinhamento de Sequência Relação Estrutura-Atividade Inibidores da Tripsina/farmacologia [Pt] Tipo de publicação: JOURNAL ARTICLE; REVIEW [Nm] Nome de substância: 0 (Anti-Bacterial Agents); 0 (Cyclotides); 0 (Insecticides); 0 (Phosphatidylethanolamines); 0 (Trypsin Inhibitors); 39382-08-6 (phosphatidylethanolamine) [Em] Mês de entrada: 1705 [Cu] Atualização por classe: 170515 [Lr] Data última revisão: 170515 [Sb] Subgrupo de revista: IM [Da] Data de entrada para processamento: 170114 [St] Status: MEDLINE [do] DOI: 10.1021/acs.biochem.6b01212

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[PMID]: 27487329 [Au] Autor: Abdul Ghani H; Henriques ST; Huang YH; Swedberg JE; Schroeder CI; Craik DJ [Ad] Endereço: Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, 4072, Australia. [Ti] Título: Structural and functional characterization of chimeric cyclotides from the Möbius and trypsin inhibitor subfamilies. [So] Source: Biopolymers;108(1), 2017 Jan. [Is] ISSN: 1097-0282 [Cp] País de publicação: United States [La] Idioma: eng [Ab] Resumo: Cyclotides are plant-derived host defense peptides displaying exceptional stability due to their cyclic cystine knot comprising three intertwined disulfide bonds and a cyclic backbone. Their six conserved cysteine residues are separated by backbone loops with diverse sequences. Prototypical cyclotides from the Möbius (kalata B1) and trypsin inhibitor (MCoTI-II) subfamilies lack sequence homology with one another, but both are able to penetrate cells, apparently via different mechanisms. To delineate the influence of the sequences of the loops on the structure and cell internalization of these two cyclotide subfamilies, a series of Möbius/trypsin inhibitor loop-chimeras of kalata B1 and MCoTI-II were synthesized, and structurally and functionally characterized. NMR analysis showed that the structural fold of the majority of chimeric peptides was minimally affected by the loop substitutions. Substituting loops 3, 5, or 6 of MCoTI-II into the corresponding loops of kalata B1 attenuated its hemolytic and cytotoxic activities, and greatly reduced its cell-penetrating properties. On the other hand, replacing loops of MCoTI-II with the corresponding loops of kalata B1 did not introduce cytotoxicity into the chimeras. Loops 2, 3, and 4 of MCoTI-II were found to contribute little to cell-penetrating properties. Overall, this study provides valuable insights into the structural basis for the hemolytic, cytotoxic, and cell-penetrating properties of kalata B1 and MCoTI-II, which could be useful for future engineering of cyclotides to carry bioactive epitopes to intracellular targets. [Mh] Termos MeSH primário: Ciclotídeos/química Proteínas de Plantas/química [Mh] Termos MeSH secundário: Sequência de Aminoácidos Sobrevivência Celular/efeitos dos fármacos Cucurbitaceae/metabolismo Ciclotídeos/síntese química Ciclotídeos/toxicidade Eritrócitos/citologia Eritrócitos/efeitos dos fármacos Eritrócitos/metabolismo Células HeLa Hemólise/efeitos dos fármacos Seres Humanos Espectroscopia de Ressonância Magnética Estrutura Terciária de Proteína [Pt] Tipo de publicação: JOURNAL ARTICLE [Nm] Nome de substância: 0 (Cyclotides); 0 (Plant Proteins); 0 (kalata B1); 0 (trypsin inhibitor MCoTI-II) [Em] Mês de entrada: 1702 [Cu] Atualização por classe: 170223 [Lr] Data última revisão: 170223 [Sb] Subgrupo de revista: IM [Da] Data de entrada para processamento: 160804 [St] Status: MEDLINE [do] DOI: 10.1002/bip.22927

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[PMID]: 28006906 [Au] Autor: Matsuura HN; Poth AG; Yendo AC; Fett-Neto AG; Craik DJ [Ad] Endereço: Center for Biotechnology and Department of Botany, Federal University of Rio Grande do Sul , Porto Alegre, RS, Brazil. [Ti] Título: Isolation and Characterization of Cyclotides from Brazilian Psychotria: Significance in Plant Defense and Co-occurrence with Antioxidant Alkaloids. [So] Source: J Nat Prod;79(12):3006-3013, 2016 Dec 23. [Is] ISSN: 1520-6025 [Cp] País de publicação: United States [La] Idioma: eng [Ab] Resumo: Plants from the genus Psychotria include species bearing cyclotides and/or alkaloids. The elucidation of factors affecting the metabolism of these molecules as well as their activities may help to understand their ecological function. In the present study, high concentrations of antioxidant indole alkaloids were found to co-occur with cyclotides in Psychotria leiocarpa and P. brachyceras. The concentrations of the major cyclotides and alkaloids in P. leiocarpa and P. brachyceras were monitored following herbivore- and pathogen-associated challenges, revealing a constitutive, phytoanticipin-like accumulation pattern. Psyleio A, the most abundant cyclotide found in the leaves of P. leiocarpa, and also found in P. brachyceras leaves, exhibited insecticidal activity against Helicoverpa armigera larvae. Addition of ethanol in the vehicle for peptide solubilization in larval feeding trials proved deleterious to insecticidal activity and resulted in increased rates of larval survival in treatments containing indole alkaloids. This suggests that plant alkaloids ingested by larvae might contribute to herbivore oxidative stress detoxification, corroborating, in a heterologous system with artificial oxidative stress stimulation, the antioxidant efficiency of Psychotria alkaloids previously observed in planta. Overall, the present study reports data for eight novel cyclotides, the identification of P. leiocarpa as a cyclotide-bearing species, and the absence of these peptides in P. umbellata. [Mh] Termos MeSH primário: Ciclotídeos/isolamento & purificação Alcaloides de Indol/isolamento & purificação Inseticidas/isolamento & purificação Inseticidas/farmacologia Psychotria/química [Mh] Termos MeSH secundário: Sequência de Aminoácidos Animais Antioxidantes/química Brasil Ciclotídeos/análise Ciclotídeos/química Ciclotídeos/farmacologia Herbivoria Alcaloides de Indol/análise Alcaloides de Indol/química Alcaloides de Indol/farmacologia Inseticidas/química Larva/efeitos dos fármacos Lepidópteros/efeitos dos fármacos Dados de Sequência Molecular Estrutura Molecular Estresse Oxidativo Folhas de Planta/química [Pt] Tipo de publicação: JOURNAL ARTICLE [Nm] Nome de substância: 0 (Antioxidants); 0 (Cyclotides); 0 (Indole Alkaloids); 0 (Insecticides) [Em] Mês de entrada: 1705 [Cu] Atualização por classe: 170503 [Lr] Data última revisão: 170503 [Sb] Subgrupo de revista: IM [Da] Data de entrada para processamento: 161224 [St] Status: MEDLINE [do] DOI: 10.1021/acs.jnatprod.6b00492

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[PMID]: 27809507 [Au] Autor: Pinto MF; Silva ON; Viana JC; Porto WF; Migliolo L; B da Cunha N; Gomes N; Fensterseifer IC; Colgrave ML; Craik DJ; Dias SC; Franco OL [Ad] Endereço: Centro de Análises Proteômicas e Bioquímicas, Programa de Pós-Graduação em Ciências Genômicas e Biotecnologia, Universidade Católica de Brasília , Brasília-DF, Brazil. [Ti] Título: Characterization of a Bioactive Acyclotide from Palicourea rigida. [So] Source: J Nat Prod;79(11):2767-2773, 2016 Nov 23. [Is] ISSN: 1520-6025 [Cp] País de publicação: United States [La] Idioma: eng [Ab] Resumo: The extraction and purification of parigidin-br3, a cyclotide analogue belonging to the "bracelet" subfamily, from Palicourea rigida leaves is discussed. Unlike conventional cyclotides, parigidin-br3 has free N- and C-termini, as identified by MALDI-TOF/TOF analysis and confirmed by gene structure elucidation, and is one of a small number of acyclotides discovered during recent years. Parigidin-br3 showed cytotoxic activity against MCF-7 (breast cancer) and CACO2 (colorectal adenocarcinoma) cells, with IC values of ∼2.5 µM and less than 10% hemolytic activity. Overall, parigidin-br3 is a promising new molecule with cytotoxic properties against tumor cell lines and, unlike many synthetic acyclic analogues, demonstrates that cytotoxic activity is not limited to conventional (i.e., cyclic) cyclotides. [Mh] Termos MeSH primário: Antineoplásicos Fitogênicos/isolamento & purificação Rubiaceae/química [Mh] Termos MeSH secundário: Sequência de Aminoácidos Antineoplásicos Fitogênicos/química Antineoplásicos Fitogênicos/farmacologia Neoplasias da Mama/tratamento farmacológico Células CACO-2 Neoplasias Colorretais/tratamento farmacológico Ciclotídeos/química Ensaios de Seleção de Medicamentos Antitumorais Feminino Seres Humanos Concentração Inibidora 50 Dados de Sequência Molecular Estrutura Molecular Folhas de Planta/química Proteínas de Plantas/química Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz [Pt] Tipo de publicação: JOURNAL ARTICLE [Nm] Nome de substância: 0 (Antineoplastic Agents, Phytogenic); 0 (Cyclotides); 0 (Plant Proteins) [Em] Mês de entrada: 1706 [Cu] Atualização por classe: 170609 [Lr] Data última revisão: 170609 [Sb] Subgrupo de revista: IM [Da] Data de entrada para processamento: 161105 [St] Status: MEDLINE

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[PMID]: 27603276 [Au] Autor: Melander E; Eriksson C; Jansson B; Göransson U; Hammarlund-Udenaes M [Ad] Endereço: Department of Pharmaceutical Biosciences, Faculty of Pharmacy, Uppsala University, Uppsala, Sweden. [Ti] Título: Improved method for quantitative analysis of the cyclotide kalata B1 in plasma and brain homogenate. [So] Source: Biopolymers;106(6):910-916, 2016 Nov. [Is] ISSN: 1097-0282 [Cp] País de publicação: United States [La] Idioma: eng [Ab] Resumo: This study provides a new method for quantifying the cyclotide kalata B1 in both plasma and brain homogenate. Cyclotides are ultra-stable peptides with three disulfide bonds that are interesting from a drug development perspective as they can be used as scaffolds. In this study we describe a new validated LC-MS/MS method with high sensitivity and specificity for kalata B1. The limit of quantification was 2 ng/mL in plasma and 5 ng/gmL in brain homogenate. The method was linear in the range 2-10,000 ng/mL for plasma and 5-2000 ng/g for brain. Liquid Chromatographic separation was performed on a HyPurity C18 column, 50 × 4.6 mm, 3 µm particle size. The method had inter- and intra-day precision and accuracy levels <15% and 12% respectively. Applying the method to in vivo plasma samples and brain homogenate samples from equilibrium dialysis yielded satisfying results and was able to describe the plasma pharmacokinetics and brain tissue binding of kalata B1. The described method is quick, reproducible and well suited to quantifying kalata B1 in biological matrices. [Mh] Termos MeSH primário: Encéfalo/metabolismo Ciclotídeos/farmacocinética Espectrometria de Massas Modelos Biológicos Plasma/metabolismo [Mh] Termos MeSH secundário: Animais Ciclotídeos/farmacologia Masculino Ratos Ratos Sprague-Dawley [Pt] Tipo de publicação: JOURNAL ARTICLE [Nm] Nome de substância: 0 (Cyclotides); 0 (kalata B1) [Em] Mês de entrada: 1702 [Cu] Atualização por classe: 170220 [Lr] Data última revisão: 170220 [Sb] Subgrupo de revista: IM [Da] Data de entrada para processamento: 160908 [St] Status: MEDLINE [do] DOI: 10.1002/bip.22984

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