RESUMO
Infectious endocarditis is a life-threatening disease, and diagnostics are urgently needed to accurately diagnose this disease especially in the case of prosthetic valve endocarditis. We show here that maltohexaose conjugated to indocyanine green (MH-ICG) can detect Staphylococcus aureus (S. aureus) infection in a rat model of infective endocarditis. The affinity of MH-ICG to S. aureus was determined and had a Km and Vmax of 5.4 µM and 3.0 X 10-6 µmol/minutes/108 CFU, respectively. MH-ICG had no detectable toxicity to mammalian cells at concentrations as high as 100 µM. The in vivo efficiency of MH-ICG in rats was evaluated using a right heart endocarditis model, and the accumulation of MH-ICG in the bacterial vegetations was 2.5 ± 0.2 times higher than that in the control left ventricular wall. The biological half-life of MH-ICG in healthy rats was 14.0 ± 1.3 minutes, and approximately 50% of injected MH-ICG was excreted into the feces after 24 hours. These data demonstrate that MH-ICG was internalized by bacteria with high specificity and that MH-ICG specifically accumulated in bacterial vegetations in a rat model of endocarditis. These results demonstrate the potential efficacy of this agent in the detection of infective endocarditis.
Assuntos
Técnicas de Imagem Cardíaca/métodos , Endocardite Bacteriana/diagnóstico por imagem , Glicoconjugados/química , Verde de Indocianina/química , Oligossacarídeos/química , Infecções Estafilocócicas/diagnóstico por imagem , Animais , Células CHO , Sobrevivência Celular/efeitos dos fármacos , Corantes/química , Corantes/farmacocinética , Cricetulus , Modelos Animais de Doenças , Endocardite Bacteriana/microbiologia , Endocardite Bacteriana/patologia , Glicoconjugados/farmacocinética , Ventrículos do Coração/diagnóstico por imagem , Ventrículos do Coração/microbiologia , Ventrículos do Coração/patologia , Humanos , Verde de Indocianina/farmacocinética , Raios Infravermelhos , Masculino , Oligossacarídeos/farmacocinética , Ratos , Ratos Sprague-Dawley , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/patologia , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/metabolismo , Staphylococcus aureus/patogenicidadeRESUMO
Despite the progress in the development of novel treatment modalities, a significant portion of patients with psoriasis remains undertreated relative to the severity of their disease. Recent evidence points to targeting the glucose transporter 1 and sugar metabolism as a novel therapeutic strategy for the treatment of psoriasis and other hyperproliferative skin diseases. In this review, we discuss glycoconjugation, an approach that facilitates the pharmacokinetics of cytotoxic molecules and ensures their preferential influx through glucose transporters. We propose pathways of glycoconjugate synthesis to increase effectiveness, cellular selectivity, and tolerability of widely used antipsoriatic drugs. The presented approach exploiting the heightened glucose requirement of proliferating keratinocytes bears the potential to revolutionize the management of psoriasis. SIGNIFICANCE STATEMENT: Recent findings concerning the fundamental role of enhanced glucose metabolism and glucose transporter 1 overexpression in the pathogenesis of psoriasis brought to light approaches that proved successful in cancer treatment. Substantial advances in the emerging field of glycoconjugation highlight the rationale for the development of glucose-conjugated antipsoriatic drugs to increase their effectiveness, cellular selectivity, and tolerability. The presented approach offers a novel therapeutic strategy for the treatment of psoriasis and other hyperproliferative skin diseases.
Assuntos
Glicoconjugados/uso terapêutico , Psoríase/tratamento farmacológico , Desenvolvimento de Medicamentos , Glucose/metabolismo , Transportador de Glucose Tipo 1/fisiologia , Glicoconjugados/biossíntese , Glicoconjugados/farmacocinética , Humanos , Psoríase/metabolismo , Distribuição TecidualRESUMO
The renal primary cilium is a small microtubule-based appendage thought to have mechano/chemosensory roles detecting changes in the fluid passing through the nephron. Mutations affecting cilium structure or function of ciliary-localized proteins result in a spectrum of diseases termed ciliopathies, with prevalent phenotypes such as the formation of renal cysts and fibrosis. While many studies have been conducted using fixed kidney sections or live imaging of cells in culture to investigate the cilium, examination in the context of a living murine kidney remains to be conducted. Previously, our lab generated the SSTR3GFP mouse to study cilium dynamics in vivo and found novel cilium behaviors that occurred following alteration of heart rate, blood pressure, and tubule flow. In this manuscript, we utilize multiple transgenic mouse models and an abdominal window imaging approach to observe primary cilia and tubule flow dynamics, immune cell movement, and renal Ca2+ signaling as it occurs in real time within a live mouse kidney. We present this window method as an approach that can be used in combination with various fluorescently labeled transgenic mice to investigate renal physiology, pathology, and function in vivo in longitudinal studies for as long as 5weeks.
Assuntos
Técnicas Biossensoriais , Sinalização do Cálcio/fisiologia , Cílios/ultraestrutura , Microscopia Intravital/métodos , Túbulos Renais/ultraestrutura , Microscopia Confocal/métodos , Animais , Pressão Sanguínea/fisiologia , Receptor 1 de Quimiocina CX3C/genética , Receptor 1 de Quimiocina CX3C/metabolismo , Carbocianinas/química , Carbocianinas/farmacocinética , Cílios/fisiologia , Corantes Fluorescentes/química , Corantes Fluorescentes/farmacocinética , Expressão Gênica , Genes Reporter , Glicoconjugados/química , Glicoconjugados/farmacocinética , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Frequência Cardíaca/fisiologia , Microscopia Intravital/instrumentação , Túbulos Renais/fisiologia , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Camundongos , Camundongos Transgênicos , Microscopia Confocal/instrumentação , Receptores CCR2/genética , Receptores CCR2/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Reologia , Proteína Vermelha FluorescenteRESUMO
The simultaneous possession of high tumor-targeting efficiency, long blood circulation, and low normal-tissue retention is critical for future clinically translatable nanomedicines. Herein, we reported a facile in situ glycoconjugation strategy for the synthesis of near-infrared (NIR)-emitting gold glyconanoparticles (AuGNPs, â¼2.4 nm) using 1-thio-ß-d-glucose as both the surface ligand and the reducing agent in the presence of a gold precursor. The ultrasmall AuGNPs showed similar low healthy organ retention to that of the renal-clearable ultrasmall nonglyconanoparticles, but â¼10 and 2.5 times higher in vitro and in vivo tumor-targeting efficiencies, respectively, were observed. This facile glycoconjugation strategy of ultrasmall AuGNPs was found to show activity towards glucose transporters in the cancer cells and prolonged blood circulation with both renal and hepatobiliary clearance pathways, which synergistically enhanced the tumor targeting of the ultrasmall AuGNPs. This discovery provides a smart strategy for the improvement in tumor targeting by ultrasmall NPs and further strengthens our understanding of glycoconjugation in designing future clinically translatable nanomedicines.
Assuntos
Sistemas de Liberação de Medicamentos , Corantes Fluorescentes , Glicoconjugados , Ouro , Nanopartículas Metálicas , Neoplasias Experimentais , Animais , Linhagem Celular Tumoral , Corantes Fluorescentes/química , Corantes Fluorescentes/farmacocinética , Corantes Fluorescentes/farmacologia , Glicoconjugados/química , Glicoconjugados/farmacocinética , Glicoconjugados/farmacologia , Ouro/química , Ouro/farmacocinética , Ouro/farmacologia , Humanos , Nanopartículas Metálicas/química , Nanopartículas Metálicas/uso terapêutico , Camundongos , Camundongos Nus , Neoplasias Experimentais/diagnóstico por imagem , Neoplasias Experimentais/tratamento farmacológico , Neoplasias Experimentais/metabolismoRESUMO
Adrenomedullin (AM) is a multifunctional bioactive peptide. Recent studies have shown that AM has protective effects against ischemic brain damage. We recently prepared a long-acting human AM derivative that was conjugated with a 60â¯kDa polyethylene glycol (PEG-AM), which had an effect similar to that of native AM. In this study, we examined the effect of PEG-AM on four-vessel occlusion model rats, which exhibit vascular dementia. From day 10 to day 14 after surgery, the learning and memory abilities of the rats were examined using a Morris water maze. The rats were treated with a single subcutaneous injection of 1.0 or 10.0â¯nmol/kg of PEG-AM. PEG-AM treatment reduced the escape latency in the hidden platform test. Furthermore, the treatment increased the time spent in the platform quadrant in the probe test. The data showed that PEG-AM injection prevented memory loss and learning disorders in dose-dependent manner. On day 14, the immunoreactive AM concentration in plasma was 9.749⯱â¯2.167â¯pM in the high-dose group (10.0â¯nmol/kg) and 0.334⯱â¯0.073â¯pM in the low-dose group (1.0â¯nmol/kg). However, even in the low-dose group, a significant effect was observed in both tests. The present data indicate that PEG-AM is a possible therapeutic agent for the treatment of ischemic brain injury or vascular dementia.
Assuntos
Adrenomedulina/farmacologia , Lesões Encefálicas/tratamento farmacológico , Demência Vascular/tratamento farmacológico , Glicoconjugados/farmacologia , Nootrópicos/farmacologia , Traumatismo por Reperfusão/tratamento farmacológico , Vasodilatadores/farmacologia , Insuficiência Vertebrobasilar/tratamento farmacológico , Adrenomedulina/química , Adrenomedulina/farmacocinética , Animais , Lesões Encefálicas/fisiopatologia , Demência Vascular/fisiopatologia , Modelos Animais de Doenças , Esquema de Medicação , Glicoconjugados/química , Glicoconjugados/farmacocinética , Humanos , Injeções Subcutâneas , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Aprendizagem em Labirinto/fisiologia , Memória/efeitos dos fármacos , Memória/fisiologia , Nootrópicos/química , Nootrópicos/farmacocinética , Polietilenoglicóis/química , Ratos , Ratos Wistar , Traumatismo por Reperfusão/fisiopatologia , Resultado do Tratamento , Vasodilatadores/química , Vasodilatadores/farmacocinética , Insuficiência Vertebrobasilar/fisiopatologiaRESUMO
Introduction: Plenty of new FVIII/IX concentrates have been developed and entered the market of hemophilia treatment. Others are going to end the long/demanding procedures for approval. Changes of the FVIII molecule (single chain), pegylation of B-domain deleted FVIII, and fusion with Fc succeeded to improve the FVIII half-life, about 4 hours. Pegylation and fusion with albumin or Fc of rFIX caused a substantial increase of half-life, approximately 3-4 times that of FIX standard concentrates. Area covered: Extended Half-life concentrates may allow a longer time interval between the prophylaxis bolus, a feature very well accepted by young patients. Also, adherence of adolescents can be improved by these new, less demanding, concentrates. The immunogenicity of these new molecules is so far under post-marketing evaluation. The incidence of neutralizing antibodies is very low in previously treated patients, but the data on previously untreated patients are not yet assessed. The cost of some Extended Half-Life concentrates is higher than that of standard ones, and some concerns have been raised about the cost for public or private health care institutions. Expert opinion: An accurate evaluation of patients' needs, individual pharmacokinetics, and cost/effectiveness might allow a more appropriate usage of these new and expensive concentrates.
Assuntos
Coagulantes/farmacocinética , Fator IX/farmacocinética , Fator VIII/farmacocinética , Glicoconjugados/farmacocinética , Hemofilia A/tratamento farmacológico , Hemofilia B/dietoterapia , Albuminas/química , Coagulantes/química , Fator IX/química , Fator VIII/química , Glicoconjugados/química , Meia-Vida , Hemofilia A/sangue , Hemofilia A/psicologia , Hemofilia B/sangue , Hemofilia B/psicologia , Humanos , Fragmentos Fc das Imunoglobulinas/química , Cooperação do Paciente , Polietilenoglicóis/química , Qualidade de Vida/psicologiaRESUMO
The synthesis and application of porphyrins has seen a huge shift towards research in porphyrin bio-molecular based systems in the past decade. The preferential localization of porphyrins in tumors, as well as their ability to generate reactive singlet oxygen and low dark toxicities has resulted in their use in therapeutic applications such as photodynamic therapy. However, their inherent lack of bio-distribution due to water insolubility has shifted research into porphyrin-nanomaterial conjugated systems to address this challenge. This has broadened their bio-applications, viz. bio-sensors, fluorescence tracking, in vivo magnetic resonance imaging (MRI), and positron emission tomography (PET)/CT imaging to photo-immuno-therapy just to highlight a few. This paper reviews the unique theranostic role of porphyrins in disease diagnosis and therapy. The review highlights porphyrin conjugated systems and their applications. The review ends by bringing current challenges and future perspectives of porphyrin based conjugated systems and their respective applications into light.
Assuntos
Glicoconjugados/síntese química , Neoplasias/diagnóstico por imagem , Neoplasias/terapia , Fármacos Fotossensibilizantes/síntese química , Porfirinas/síntese química , Animais , Glicoconjugados/farmacocinética , Glicoconjugados/farmacologia , Humanos , Imageamento por Ressonância Magnética/métodos , Camundongos , Neoplasias/patologia , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacocinética , Fármacos Fotossensibilizantes/farmacologia , Porfirinas/farmacocinética , Porfirinas/farmacologia , Tomografia por Emissão de Pósitrons/métodos , Oxigênio Singlete/química , Oxigênio Singlete/metabolismo , Nanomedicina Teranóstica/métodos , Água/química , Água/metabolismoRESUMO
Drug delivery carriers are now widely established because they can increase the therapeutic efficiency of drugs. In general, the aim in this field is to create effective carriers that have large amounts of drugs loaded to minimize drug carrier material that needs to be disposed of. However, there has been little attention so far in the literature on the effect of the amount of loaded drugs on the biological activity. In this paper, we are trying to answer the question of how the drug-loading content will affect the in vitro activity. We use two methods to load paclitaxel (PTX) into micelles based on the glycopolymer, poly(1- O-methacryloyl-ß-d-fructopyranose)- block-poly(methyl methacylate) (Poly(1- O-MAFru)35- b-PMMA145). In the one-step method, the drug is loaded into the particles during the self-assembly process. However, the size of nanoparticle increased with the PTX content from 26 to 50 nm, triggering enhanced cellular uptake by MCF-7 and MDA-MB-231, which was caused by changes in diameter size and not by changes in drug concentration. To keep the nanoparticle size constant, preformed micelles were loaded with PTX (two-step process). The increasing amount of loaded drug led to decreased cellular uptake and reduced cytotoxicity by the cancer cell lines. Small-angle neutron scattering and small-angle X-ray scattering, supported by transmission electron microscopy and dynamic light scattering, exposed the PTX location in the shell. This caused shrinkage of the shell and lower levels of shell hydration, resulting in lower cellular uptake and lower cytotoxicity. Upon the release of PTX, the shell regained its original level of hydration. We could show that because drug loading causes morphology changes, in either the shell or the size, it is impossible to separate the parameters that will influence the biological activity. Although the same phenomenon may not apply to every drug delivery system, it needs to be considered that except for the well-known parameters that affect cell uptake-size, shape, surface chemistry, type of nanoparticle, and presence of bioactive groups-the amount of loaded drugs might change the physicochemical parameters of the nanoparticle and thus the in vitro and potentially the in vivo outcomes.
Assuntos
Portadores de Fármacos , Glicoconjugados , Micelas , Nanopartículas/química , Paclitaxel , Portadores de Fármacos/síntese química , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Portadores de Fármacos/farmacologia , Glicoconjugados/síntese química , Glicoconjugados/química , Glicoconjugados/farmacocinética , Glicoconjugados/farmacologia , Humanos , Células MCF-7 , Paclitaxel/química , Paclitaxel/farmacocinética , Paclitaxel/farmacologia , Tamanho da PartículaRESUMO
Breast cancer, up-regulated with human epidermal growth factor receptor type-2 (HER-2) has led to the concept of developing HER-2 targeted anticancer therapeutics. Docetaxel-loaded D-α-tocopherol polyethylene glycol 1000 succinate conjugated chitosan (TPGS-g-chitosan) nanoparticles were prepared with or without Trastuzumab decoration. The particle size and entrapment efficiency of conventional, non-targeted as well as targeted nanoparticles were in the range of 126-186 nm and 74-78% respectively. In-vitro studies on SK-BR-3 cells showed that docetaxel-loaded non-targeted and HER-2 receptor targeted TPGS-g-chitosan nanoparticles have enhanced the cellular uptake and cytotoxicity with a promising bioadhesion property, in comparison to conventional formulation i.e., Docel™. The IC50 values of non-targeted and targeted nanoparticles from cytotoxic assay were found to be 43 and 223 folds higher than Docel™. The in-vivo pharmacokinetic study showed 2.33, and 2.82-fold enhancement in relative bioavailability of docetaxel for non-targeted and HER-2 receptor targeted nanoparticles, respectively than Docel™. Further, after i.v administration, non-targeted and targeted nanoparticles achieved 3.48 and 5.94 times prolonged half-life in comparison to Docel™. The area under the curve (AUC), relative bioavailability (FR) and mean residence time (MRT) were found to be higher for non-targeted and targeted nanoparticles when compared to Docel™. The histopathology studies of non-targeted and targeted nanoparticles showed less toxicity on vital organs such as lungs, liver, and kidney when compared to Docel™.
Assuntos
Adenocarcinoma/tratamento farmacológico , Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Docetaxel/farmacologia , Glicoconjugados/farmacologia , Trastuzumab/farmacologia , Vitamina E/farmacologia , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Animais , Antineoplásicos/química , Antineoplásicos/farmacocinética , Disponibilidade Biológica , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Quitosana/química , Docetaxel/química , Docetaxel/farmacocinética , Portadores de Fármacos , Feminino , Glicoconjugados/química , Glicoconjugados/farmacocinética , Humanos , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Terapia de Alvo Molecular , Nanopartículas/química , Nanopartículas/ultraestrutura , Ratos , Receptor ErbB-2/metabolismo , Trastuzumab/química , Trastuzumab/farmacocinética , Carga Tumoral/efeitos dos fármacos , Vitamina E/química , Vitamina E/farmacocinética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Carbohydrate receptors on liver represent attractive targets for receptor-mediated delivery of nanostructured therapeutics. In this study, two new cholesterol-based glycoconjugates derived from d-galactose and N-acetylglucosamine were synthesized and incorporated into liposomes. 99mTc-Cholesterol-DTPA complex was used for radiolabeling experiments in vivo with high radiochemical yields and stability. Biodistribution studies confirmed the targeting of galactosylated liposomes (GalL) to liver cells. These results indicated that GalL could be considered a promising drug delivery system for liver diseases therapy.
Assuntos
Colesterol/uso terapêutico , Glicoconjugados/uso terapêutico , Hepatopatias/tratamento farmacológico , Administração Intravenosa , Animais , Colesterol/administração & dosagem , Colesterol/farmacocinética , Sistemas de Liberação de Medicamentos , Feminino , Glicoconjugados/administração & dosagem , Glicoconjugados/farmacocinética , Lipossomos/administração & dosagem , Lipossomos/farmacocinética , Lipossomos/uso terapêutico , Hepatopatias/sangue , Camundongos , Camundongos Endogâmicos BALB C , Distribuição TecidualRESUMO
This work represents the first broad study of testing diverse heterogenous glycoconjugates (7 different glycoalbumins) for their differential in vivo binding (11 different cancer cell types) in both cell- and animal-based studies. As a result, various changes in biodistribution, excretion, and even tumor adhesion were observed.
Assuntos
Moléculas de Adesão Celular/farmacocinética , Glicoconjugados/farmacocinética , Albumina Sérica/farmacocinética , Animais , Moléculas de Adesão Celular/química , Linhagem Celular Tumoral , Produtos Finais de Glicação Avançada , Glicoconjugados/química , Humanos , Camundongos , Especificidade de Órgãos , Albumina Sérica/química , Distribuição Tecidual , Albumina Sérica GlicadaRESUMO
Nanotechnology, focused on discovery and development of new pharmaceutical products is known as nanopharmacology, and one research area this branch is engaged in are nanopharmaceuticals. The importance of being nano has been particularly emphasized in scientific areas dealing with nanomedicine and nanopharmaceuticals. Nanopharmaceuticals, their routes of administration, obstacles and solutions concerning their improved application and enhanced efficacy have been briefly yet comprehensively described. Cancer is one of the leading causes of death worldwide and evergrowing number of scientific research on the topic only confirms that the needs have not been completed yet and that there is a wide platform for improvement. This is undoubtedly true for nanoformulations of an anticancer drug doxorubicin, where various nanocarrriers were given an important role to reduce the drug toxicity, while the efficacy of the drug was supposed to be retained or preferably enhanced. Therefore, we present an interdisciplinary comprehensive overview of interdisciplinary nature on nanopharmaceuticals based on doxorubicin and its nanoformulations with valuable information concerning trends, obstacles and prospective of nanopharmaceuticals development, mode of activity of sole drug doxorubicin and its nanoformulations based on different nanocarriers, their brief descriptions of biological activity through assessing in vitro and in vivo behavior.
Assuntos
Antibióticos Antineoplásicos/farmacologia , Doxorrubicina/farmacologia , Nanomedicina/métodos , Nanopartículas/química , Nanotecnologia/métodos , Neoplasias/tratamento farmacológico , Animais , Antibióticos Antineoplásicos/química , Antibióticos Antineoplásicos/farmacocinética , Apoptose/efeitos dos fármacos , Transporte Biológico , Linhagem Celular Tumoral , Ceramidas/agonistas , Ceramidas/metabolismo , Química Farmacêutica , Modelos Animais de Doenças , Doxorrubicina/química , Doxorrubicina/farmacocinética , Portadores de Fármacos/síntese química , Portadores de Fármacos/farmacocinética , Glicoconjugados/síntese química , Glicoconjugados/farmacocinética , Humanos , Nanopartículas/administração & dosagem , Neoplasias/metabolismo , Neoplasias/patologia , Polietilenoglicóis/síntese química , Polietilenoglicóis/farmacocinética , Espécies Reativas de Oxigênio/agonistas , Espécies Reativas de Oxigênio/metabolismoRESUMO
Novel cis-2-methylmalonato(trans-R,R-cyclohexane-1,2-diamine)platinum(II) glycoconjugates derived from different sugar motifs, namely, glucose (Glu-Me-Pt), mannose (Man-Me-Pt) and galactose (Gal-Me-Pt) were designed and synthesized based on the third generation clinical drug oxaliplatin for potential glucose transporters (GLUTs) mediated tumor targeting. All platinum(II) glycoconjugates were characterized by 1H NMR, 13C NMR, IR, HRMS as well as 195Pt-NMR analysis. Despite their substantial improvement in water solubility, the conjugates exhibited comparable or better in vitro cytotoxicities than oxaliplatin determined in six different human cancer cell lines. Glu-Me-Pt has been shown to be more effective than cisplatin and oxaliplatin with improved therapeutic index in leukemia-bearing DBA/2 mice model. The potential GLUT transportability of the complexes was investigated using cell-based fluorescent competition assay and GLUT inhibitor mediated cell viability analysis in GLUT over-expressing HT29 cell line. Each sugar motif was found to be useful to enable the platinum(II) complexes as substrate for GLUT mediated cell uptake. In vitro DNA adduct formation analysis has been investigated for the first time for this class of compounds to reveal the intrinsic differences in antitumor activity between the malonatoplatinum(II) glycoconjugates and oxaliplatin. The intrinsic DNA reactivity of the platinum(II)-sugar conjugates was found as Gal-Me-Pt > Glu-Me-Pt > Man-Me-Pt ≈ oxaliplatin by kinetic study on the formation of platinum(II) adducts with guanosine-5'-monophosphate (5'-GMP). The results from this study demonstrate the usefulness of glucose, mannose and galactose as alternative sugar motif on glycoconjugation for GLUT mediated drug design and pharmaceutical R&D, and the obtained fundamental results also support the potential of the GLUT targeted platinum(II)-sugar conjugates as lead compounds for further pre-clinical evaluation.
Assuntos
Antineoplásicos/química , Antineoplásicos/farmacologia , Glicoconjugados/química , Glicoconjugados/farmacologia , Compostos Organoplatínicos/química , Compostos Organoplatínicos/farmacologia , Animais , Antineoplásicos/farmacocinética , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Cisplatino/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Proteínas Facilitadoras de Transporte de Glucose/metabolismo , Glicoconjugados/farmacocinética , Glicoconjugados/uso terapêutico , Humanos , Masculino , Metilação , Camundongos , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Compostos Organoplatínicos/farmacocinética , Compostos Organoplatínicos/uso terapêutico , OxaliplatinaRESUMO
Two novel fluorescent bioprobes, namely, 6N-Gly-Cy3 and 6N-Gly-Cy5, were designed and synthesized for real-time glucose transport imaging as well as potentially useful tracer for galactokinase metabolism. The structure of the bioprobes was fully characterized by 1H NMR, 13C NMR, IR, and HRMS. The fluorescence properties, glucose transporter (GLUT) specificity, and the quenching and safety profiles were studied. The cellular uptake of both bioprobes was competitively diminished by d-glucose, 2-deoxy-d-glucose and GLUT specific inhibitor in a dose-dependent manner in human colon cancer cells (HT29). Comparison study results revealed that the 6N-derived bioprobes are more useful for real-time imaging of cell-based glucose uptake than the structurally similar fluorescent tracer 6-NBDG which was not applicable under physiological conditions. The up to 96 h long-lasting quenching property of 6N-Gly-Cy5 in HT29 suggested the potential applcability of the probe for cell labeling in xenograft transplantation as well as in vivo animal imaging studies.
Assuntos
Carbocianinas/farmacocinética , Proteínas Facilitadoras de Transporte de Glucose/metabolismo , Microscopia de Fluorescência/métodos , Imagem Molecular/métodos , Carbocianinas/síntese química , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/farmacocinética , Glicoconjugados/síntese química , Glicoconjugados/farmacocinética , Células HT29 , Humanos , Espectrometria de Fluorescência/métodosRESUMO
Triantennary N-acetyl galactosamine (GalNAc3)-conjugated antisense oligonucleotides (ASOs) have greatly improved potency due to receptor-mediated uptake into hepatocyte. The disposition and pharmacokinetics of ISIS 681257, a GalNAc3-conjugated ASO, were studied in monkeys. Following subcutaneous (SC) injection, ISIS 681257 was rapidly absorbed into the systemic circulation, with peak plasma levels observed within hours after dosing. After reaching Cmax, plasma concentrations rapidly declined in a multiexponential manner and were characterized by a dominant initial rapid distribution phase in which drug transferred to tissues from circulation, followed by a much slower terminal elimination phase (half-life of 4 weeks). Intact ISIS 681257 is the major full-length oligonucleotide species in plasma (≥70%). In tissues, the conjugated-GalNAc sugar moiety was rapidly metabolized, leaving the fully unconjugated form as the only full-length oligonucleotide detected at 48 h after dosing. Unconjugated ISIS 681257 cleared slowly from tissues with a half-life of 4 weeks. ISIS 681257 was highly bound to plasma proteins (>97% bound), which limited its urinary excretion. Disposition of ISIS 681257 in plasma and liver appeared nonlinear over the 1-40 mg/kg dose range studied. The plasma and liver tissue concentration data were well described by a population based mixed-effects modeling approach with Michaelis-Menten uptake from plasma to liver. Safety data from the study and the good exposure, as well as the extended half-life of the unconjugated ASO in the liver, support further development and less frequent dosing in Phase I clinical study.
Assuntos
Acetilgalactosamina/farmacocinética , Glicoconjugados/farmacocinética , Lipoproteína(a)/metabolismo , Oligonucleotídeos Antissenso/farmacocinética , Oligonucleotídeos Fosforotioatos/farmacocinética , Acetilgalactosamina/metabolismo , Animais , Biotransformação , Proteínas Sanguíneas/metabolismo , Feminino , Glicoconjugados/metabolismo , Meia-Vida , Hepatócitos/metabolismo , Injeções Subcutâneas , Fígado/metabolismo , Masculino , Oligonucleotídeos Antissenso/metabolismo , Oligonucleotídeos Fosforotioatos/metabolismo , Ligação Proteica , Clivagem do RNARESUMO
Herein the biodistributions and in vivo kinetics of chemically prepared neoglycoproteins are reviewed. Chemical methods can be used to conjugate various mono- and oligosaccharides onto a protein surface. The kinetics and organ-specific accumulation profiles of these glycoconjugates, which are introduced through intravenous injections, have been analyzed using conventional dissection studies as well as noninvasive methods such as single photon emission computed tomography, positron emission tomography and fluorescence imaging. These studies suggest that glycan-dependent protein distribution kinetics may be useful for pharmacological and diagnostic applications.
Assuntos
Glicoconjugados/química , Glicoproteínas/química , Monossacarídeos/química , Oligossacarídeos/química , Aminação , Animais , Sequência de Carboidratos , Cisteína/química , Glicoconjugados/metabolismo , Glicoconjugados/farmacocinética , Glicoproteínas/metabolismo , Glicoproteínas/farmacocinética , Glicosilação , Injeções Intravenosas , Lisina/química , Camundongos , Monossacarídeos/metabolismo , Oligossacarídeos/metabolismo , Imagem Óptica/métodos , Oxirredução , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Tomografia Computadorizada com Tomografia Computadorizada de Emissão de Fóton Único/métodosRESUMO
BACKGROUND: The aim of this research was to study the distribution and changes of glycoconjugates particularly their terminal sugars by using lectin histochemistry during mouse spinal cord development. METHODS: Formalin-fixed sections of mouse embryo (10-16 fetal days) were processed for lectin histochemical method. In this study, two groups of horseradish peroxidase-labeled specific lectins were used: N-acetylgalactosamine, including Dolichos biflorus, Wisteria floribunda agglutinin (WFA), Vicia villosa, Glycine max as well as focuse-binding lectins, including tetragonolobus, Ulex europaeus, and Orange peel fungus (OFA). All sections were counterstained with alcian blue (pH 2.5). RESULTS: Our results showed that only WFA and OFA reacted strongly with the floor plate cells from early to late embryonic period of developing spinal cord. The strongest reactions were related to the 14, 15, and 16 days of tissue sections incubated with OFA and WFA lectins. CONCLUSION: The present study demonstrated that cellular and molecular differentiation of the spinal cord organizers is a wholly regulated process, and α-L-fucose, α-D-GalNAc, and α/ß-D-GalNAc terminal sugars play a significant role during the prenatal spinal cord development.
Assuntos
Acetilgalactosamina/metabolismo , Fucose/metabolismo , Glicoconjugados/metabolismo , Lectinas de Plantas/metabolismo , Medula Espinal/embriologia , Animais , Embrião de Mamíferos/embriologia , Feminino , Glicoconjugados/farmacocinética , Lectinas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Neurulação/fisiologia , Receptores de N-Acetilglucosamina/metabolismo , Medula Espinal/metabolismo , Fixação de TecidosRESUMO
A new tridentate bifunctional chelator, N-(-2-picolyl)(-4-hydroxy)(-3-amino)benzoic acid (PHAB), was designed to efficiently coordinate the [(99m)Tc(CO)3](+) core and facilitate coupling reactions to biomolecules. The chelator can be procured in the form of the corresponding benzotriazole ester (PHAB-OBT), which can be stored and used as a bioconjugation kit. PHAB-OBT reacts with modified carbohydrates with high selectivity and efficiency in a single step in both aqueous and organic media. As is desirable for a kit, no complicated chemical bench work is required. Glycoconjugate postlabeling resulted in neutral radiolabeled glycans with high radiochemical yields. Prelabeling approaches were assessed by successive reaction of PHAB-OBT with the [(99m)Tc(CO)3](+) core and a modified galactose model. The radiolabeled galactose was obtained in 84% yield as defined by HPLC analysis. Biodistribution of the radioactive (99m)Tc-labeled chelator, as well as the glycoconjugates, were examined in mice. Noticeably different biodistribution patterns were observed that reflect trends in the uptake of carbohydrate analogues by various organs.
Assuntos
Quelantes/química , Glicoconjugados/química , Animais , Cristalografia por Raios X , Galactose/química , Glicoconjugados/síntese química , Glicoconjugados/farmacocinética , Marcação por Isótopo , Camundongos , Camundongos Endogâmicos BALB C , Conformação Molecular , Compostos de Organotecnécio/química , Solventes/química , Distribuição Tecidual , Tomografia Computadorizada de Emissão de Fóton Único , Triazóis/químicaRESUMO
Recombinant gut hormone oxyntomodulin (OXM) is known to act as a satiety signal in human subjects and has therapeutic potential as an appetite controlling agent. The only form of this hormone that has a prospective use is a modified one, because native OXM has a very short half-life in vivo. Conjugation of OXM and the natural hydrophilic polymer polysialic acid (PSA) may significantly improve its half-life. Chemical polysialylation in vitro was used to create a long-acting form of OXM, the polysialic acid-oxyntomodulin (PSA-OXM) conjugate. The conjugation site was identified using mass shift comparative analysis of Asp-N proteolytic digests. The anorexic effect of the conjugate was tested on the lean, fasted mouse model. A two-stage purification technique was developed to obtain a homogeneous PSA-OXM conjugate, suitable for in vivo testing. The N-terminal backbone primary amino group was found to be the only point of conjugation. The conjugate obtained was resistant to the DPP-IV protease. A single injection of PSA-OXM at 15 µmol/kg dose was sufficient to maintain a steady decrease in food consumption for 8 h (P < 0.05). The length of the anorexic effect achieved is comparable to other long-acting derivatives of OXM but it requires a much higher dose for administration. It is expected that site-directed attachment of the PSA chain to the inner residues of OXM, away from the site of interaction with receptors, would produce a compound with a higher specific activity but comparable stability in the bloodstream. The conjugation technique used may be used to create OXM derivatives and other related hormones to obtain long-lasting variants, with improved suitability for clinical use.
Assuntos
Depressores do Apetite/síntese química , Ingestão de Alimentos/efeitos dos fármacos , Glicoconjugados/síntese química , Oxintomodulina/síntese química , Ácidos Siálicos/química , Animais , Depressores do Apetite/farmacocinética , Depressores do Apetite/farmacologia , Dipeptidil Peptidase 4/metabolismo , Desenho de Fármacos , Glicoconjugados/farmacocinética , Glicoconjugados/farmacologia , Meia-Vida , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Oxintomodulina/farmacocinética , Oxintomodulina/farmacologia , Fragmentos de Peptídeos/análise , ProteóliseRESUMO
We report the synthesis of bioconjugated zinc porphyrin dimers 1a-e designed as photosensitizers for one-photon and two-photon excited photodynamic therapy. These macrocycles are substituted with carbohydrate units (glucose, mannose, lactose) in order to target tumor cells over-expressing lectin membrane receptors. Polarity, singlet oxygen production and in vitro photocytotoxicity are studied to determine their photodynamic therapy potentiality.