RESUMEN
A new series of (2S,3R,4R,5S,6R)-5-fluoro-6-(hydroxymethyl)-2-aryltetrahydro-2H-pyran-3,4-diols as dual inhibitors of sodium glucose co-transporter proteins (SGLTs) were disclosed. Two methods were developed to efficiently synthesize C5-fluoro-lactones 3 and 4, which are key intermediates to the C5-fluoro-hexose based C-aryl glucosides. Compound 2b demonstrated potent hSGLT1 and hSGLT2 inhibition (IC50â¯=â¯43â¯nM for SGLT1 and IC50â¯=â¯9â¯nM for SGLT2). It showed robust inhibition of blood glucose excursion in oral glucose tolerance test (OGTT) in Sprague Dawley (SD) rats and exerted pronounced antihyperglycemic effects in db/db mice and high-fat diet-fed ZDF rats when dosed orally at 10â¯mg/kg.
Asunto(s)
Desoxiglucosa/uso terapéutico , Diabetes Mellitus Experimental/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Transportador 1 de Sodio-Glucosa/antagonistas & inhibidores , Inhibidores del Cotransportador de Sodio-Glucosa 2/uso terapéutico , Administración Oral , Animales , Glucemia/efectos de los fármacos , Desoxiglucosa/administración & dosificación , Desoxiglucosa/análogos & derivados , Desoxiglucosa/síntesis química , Diseño de Fármacos , Humanos , Hipoglucemiantes/administración & dosificación , Hipoglucemiantes/síntesis química , Hipoglucemiantes/química , Macaca fascicularis , Masculino , Ratones , Microsomas Hepáticos/metabolismo , Estructura Molecular , Ratas Sprague-Dawley , Ratas Zucker , Transportador 2 de Sodio-Glucosa/metabolismo , Inhibidores del Cotransportador de Sodio-Glucosa 2/administración & dosificación , Inhibidores del Cotransportador de Sodio-Glucosa 2/síntesis química , Inhibidores del Cotransportador de Sodio-Glucosa 2/química , Relación Estructura-ActividadRESUMEN
Metabolic chemical reporters of glycosylation in combination with bioorthogonal reactions have been known for two decades and have been used by many different research laboratories for the identification and visualization of glycoconjugates. More recently, however, they have begun to see utility for the investigation of cellular metabolism and the tolerance of biosynthetic enzymes and glycosyltransferases to different sugars. Here, we take this concept one step further by using the metabolic chemical reporter 6-azido-6-deoxy-glucose (6AzGlc). We show that treatment of mammalian cells with the per- O-acetylated version of 6AzGlc results in robust labeling of a variety of proteins. Notably, the pattern of this labeling was consistent with O-GlcNAc modifications, suggesting that the enzyme O-GlcNAc transferase is quite promiscuous for its donor sugar substrates. To confirm this possibility, we show that 6AzGlc-treatment results in the labeling of known O-GlcNAcylated proteins, that the UDP-6AzGlc donor sugar is indeed produced in living cells, and that recombinant OGT will accept UDP-6AzGlc as a substrate in vitro. Finally, we use proteomics to first identify several bona fide 6AzGlc-modifications in mammalian cells and then an endogenous O-glucose modification on host cell factor. These results support the conclusion that OGT can endogenously modify proteins with both N-acetyl-glucosamine and glucose, raising the possibility that intracellular O-glucose modification may be a widespread modification under certain conditions or in particular tissues.
Asunto(s)
Azidas/metabolismo , Desoxiglucosa/análogos & derivados , Desoxiglucosa/metabolismo , N-Acetilglucosaminiltransferasas/metabolismo , Proteínas/metabolismo , Animales , Azidas/síntesis química , Azidas/química , Línea Celular Tumoral , Chlorocebus aethiops , Desoxiglucosa/síntesis química , Glicosilación , Humanos , Ratones , Procesamiento Proteico-Postraduccional , Especificidad por Sustrato , Azúcares de Uridina Difosfato/biosíntesis , beta-N-Acetilhexosaminidasas/metabolismoRESUMEN
Malignant neoplasms exhibit an elevated rate of glycolysis and a high demand for glucose over normal cells. This characteristic can be exploited for in vivo imaging and tumor targeting examined. In this manuscript, we describe the synthesis of near-infrared (NIR) fluorochrome IR-822-labeled 2-amino-2-deoxy-d-glucose (DG) for optical imaging of tumors in mice. NIR fluorescent dye IR-820 was subsequently conjugated with 3-Mercaptopropionic acid and 2-amino-2-deoxy-d-glucose to form IR-822-DG. The cell experiments and acute toxicity studies demonstrated the low toxicity of IR-822-DG to normal cells/tissues. The dynamic behavior and targeting ability of IR-822-DG in normal mice was investigated with a NIR fluorescence imaging system. The in vitro and in vivo tumor targeting capabilities of IR-822-DG were evaluated in tumor cells and tumor bearing mice, respectively. Results demonstrated that IR-822-DG actively and efficiently accumulated at the site of the tumor. The probe also exhibited good photostability and excellent cell membrane permeability. The study indicates the broad applicability of IR-822-DG for tumors diagnosis, especially in the glucose-related pathologies.
Asunto(s)
Desoxiglucosa/química , Colorantes Fluorescentes/química , Neoplasias/diagnóstico , Animales , Línea Celular Tumoral , Desoxiglucosa/síntesis química , Colorantes Fluorescentes/síntesis química , Humanos , Ratones , Ratones Desnudos , Estructura MolecularRESUMEN
Mono-, di- and trisaccharide derivatives of 1,2-unsaturated N-acetyl-d-glucal have been synthesized and shown to function as tight-binding inhibitors/slow substrates of representative hexosaminidases. Turnover is slow and not observed in the thioamide analogue, allowing determination of the 3-dimensional structure of the complex. Inhibition is insensitive to pH and to mutation of key catalytic residues, consistent with the uncharged character of the inhibitor. These properties could render this inhibitor class less prone to development of resistance.
Asunto(s)
Desoxiglucosa/análogos & derivados , Inhibidores Enzimáticos/farmacología , Hexosaminidasas/antagonistas & inhibidores , Sitios de Unión/efectos de los fármacos , Biocatálisis , Desoxiglucosa/síntesis química , Desoxiglucosa/química , Desoxiglucosa/farmacología , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Hexosaminidasas/metabolismo , Concentración de Iones de Hidrógeno , Modelos Moleculares , Estructura MolecularRESUMEN
P1-[11-(Anthracen-9-ylmethoxy)undecyl]-P2-(2-acetamido-2-deoxy-α-D-glucopyranosyl) diphosphate, a fluorescent derivative of undecyl diphosphate 2-acetamido-2-deoxyglucose, was chemically synthesized. The ability of the compound to serve as acceptor substrate of D-rhamnose residue in the enzymatic reaction catalyzed by D-rhamnosyltransferase from Pseudomonas aeruginosa PAO1 was demonstrated.
Asunto(s)
Proteínas Bacterianas/química , Desoxiglucosa , Hexosiltransferasas/química , Pseudomonas aeruginosa/enzimología , Catálisis , Desoxiglucosa/análogos & derivados , Desoxiglucosa/síntesis química , Desoxiglucosa/químicaRESUMEN
This work reports a modular and rapid approach to the stereoselective synthesis of a variety of α- and ß-(1â2)-linked C-disaccharides. The key step is a Ni-catalyzed cross-coupling reaction of D-glucal pinacol boronate with alkyl halide glycoside easily prepared from commercially available D-glucal. The products of this sp(2) -sp(3) cross-coupling reaction can be converted to glucopyranosyl, mannopyranosyl, or 2-deoxy-glucopyranosyl C-mannopyranosides by one- or two-step stereoselective oxidative-reductive transformations. To the best of our knowledge, we demonstrated the first synthetic application of a challenging sp(2) -sp(3) Suzuki-Miyaura cross-coupling reaction in carbohydrate chemistry.
Asunto(s)
Disacáridos/síntesis química , Glicósidos/síntesis química , Ácidos Borónicos/síntesis química , Ácidos Borónicos/química , Desoxiglucosa/análogos & derivados , Desoxiglucosa/síntesis química , Desoxiglucosa/química , Disacáridos/química , Glicósidos/química , Oxidación-Reducción , EstereoisomerismoRESUMEN
6-O-Tosyl-d-glucal 1 upon treatment with excess LiAlH4 unexpectedly gave 3,6-anhydro-d-glucal 2 as a major product in good yield. A crystal structure was obtained. Reaction of the anhydride 2 with N-iodosuccinimide (NIS) in excess methanol resulted in the formation of diastereomeric 2-deoxy-2-iodoglycosides. Addition of ceric (IV) ammonium nitrate and thiophenol to a solution of 2 in acetonitrile gave a mixture of 2-deoxy and 2,3-unsaturated thioglycosides. Reaction of 1,2:3,4-di-O-isopropylidine-α-d-galactopyranose with the anhydro sugar 2 in the presence of N-iodosuccinimide did not give the expected iodoglycoside mixture, but instead gave an unusual 1,4:3,6-dianhydride 7 as the major product.
Asunto(s)
Desoxiglucosa/análogos & derivados , Desoxiglucosa/síntesis química , Desoxiglucosa/química , Modelos Moleculares , Conformación MolecularRESUMEN
This study reports the radiosynthesis of a new fluorine-18 glycosylated 'click' cyanoquinoline [(18) F]5 for positron emission tomography imaging of epidermal growth factor receptor (EGFR). The tracer was obtained in 47.7 ± 7.5% (n = 3) decay-corrected radiochemical yield from 2-[(18) F]fluoro-2-deoxy-ß-d-glucopyranosyl azide, and the overall nondecay-corrected radiochemical yield from aqueous fluoride was 8.6 ± 2.3% (n = 3). An in vitro preliminary cellular uptake study showed selectivity of the tracer for EGFR-positive A431 cell lines versus EGFR-negative MCF-7 cell lines. [(18) F]5 tracer uptake in A431 cells was significantly reduced by addition of the cold isotope analogue compound 5.
Asunto(s)
Azidas/síntesis química , Desoxiglucosa/análogos & derivados , Receptores ErbB/metabolismo , Radioisótopos de Flúor/química , Quinolinas/síntesis química , Radiofármacos/síntesis química , Azidas/farmacología , Desoxiglucosa/síntesis química , Desoxiglucosa/farmacología , Humanos , Marcaje Isotópico , Células MCF-7 , Unión Proteica , Quinolinas/farmacología , Radiofármacos/farmacologíaRESUMEN
A series of tolyl 2-azido-2-deoxy-thio-glucoside donors with different combinations of protecting groups were prepared. These donors were used in glycosylation reactions to test the correlations between the stereoselectivity and the pattern of the protecting groups. Acetyl groups showed a position dependent stereo-directing effect. A remote participating mechanism is proposed to explain the observed results.
Asunto(s)
Azidas/química , Desoxiglucosa/química , Azidas/síntesis química , Conformación de Carbohidratos , Desoxiglucosa/análogos & derivados , Desoxiglucosa/síntesis química , EstereoisomerismoRESUMEN
Altered carbohydrate metabolism in cancer cells was first noted by Otto Warburg more than 80 years ago. Upregulation of genes controlling the glycolytic pathway under normoxia, known as the Warburg effect, clearly differentiates malignant from non-malignant cells. The resurgence of interest in cancer metabolism aims at a better understanding of the metabolic differences between malignant and non-malignant cells and the creation of novel therapeutic and diagnostic agents exploiting these differences. Modified d-glucose and d-mannose analogs were shown to interfere with the metabolism of their respective monosaccharide parent molecules and are potentially clinically useful anticancer and diagnostic agents. One such agent, 2-deoxy-d-glucose (2-DG), has been extensively studied in vitro and in vivo and also clinically evaluated. Studies clearly indicate that 2-DG has a pleiotropic mechanism of action. In addition to effectively inhibiting glycolysis, 2-DG has also been shown to affect protein glycosylation. In order to better understand its molecular mechanism of action, we have designed and synthesized deuterated molecular probes to study 2-DG interference with d-glucose and d-mannose metabolism using mass spectrometry. We present here the synthesis of all desired probes: 2-deutero-d-glucose, 2-deutero-d-mannose, 6-deutero-d-glucose, 6-deutero-d-mannose, and 2-deutero-2-deoxy-d-glucose as well as their complete chemical characterization.
Asunto(s)
Desoxiglucosa/química , Desoxiglucosa/síntesis química , Deuterio/química , Glucosa/química , Glucosa/síntesis química , Manosa/química , Manosa/síntesis química , Estructura MolecularRESUMEN
Two novel nonisosteric UDP-Gal analogues, (2-deoxy-2-fluoro- and 4-deoxy-4-fluoro-α-D-galactopyranosyl) phosphonoyl phosphates, were synthesized by optimized multistep procedures starting from 3,4,6-tri-O-benzyl-D-galactal and allyl 2,3,6-tri-O-benzyl-α-D-glucopyranoside, respectively. The key steps were a Michaelis-Arbuzov reaction of respective deoxy-fluoro-D-galactopyranosyl acetate with triethyl phosphite followed by a Moffatt-Khorana coupling reaction with UMP-morpholidate. The structure of all new compounds was confirmed by NMR and mass spectroscopies..
Asunto(s)
Desoxiglucosa/análogos & derivados , Organofosfonatos/síntesis química , Uridina Difosfato Galactosa/análogos & derivados , Uridina Difosfato Galactosa/síntesis química , Conformación de Carbohidratos , Desoxiglucosa/síntesis química , Desoxiglucosa/química , Organofosfonatos/química , Uridina Difosfato Galactosa/químicaRESUMEN
New syntheses of densely functionalized protected derivatives of 3-amino-3,6-dideoxyaminosugars have been accomplished in an efficient and straightforward manner. The key step of such approaches involves a highly stereoselective titanium-mediated aldol addition of a chiral α-bromo ketone, easily available from lactate esters, to crotonaldehyde. Further functional group transformations, including a new regioselective Staudinger-aza-Wittig reaction of an azidodiacetate, afford in a few steps and high yield the desired carbohydrates as advanced intermediates capable of participating in subsequent glycosylation reactions.
Asunto(s)
Amino Azúcares/síntesis química , Aldehídos/química , Amino Azúcares/química , Desoxiglucosa/análogos & derivados , Desoxiglucosa/síntesis química , Hexosaminas/síntesis química , Hexosaminas/química , Estereoisomerismo , Titanio/químicaRESUMEN
The de novo synthesis of carbohydrates constitutes an important aspect of organic chemistry, and its application toward deoxy sugars is particularly noteworthy in targeting biologically active compounds. The enantioselective preparation of 4-deoxy-D-ribo-, 4-deoxy-D-lyxo-, and 4-deoxy-D-xylo-hexopyranosides, along with their uronate counterparts has been successfully accomplished using hetero-Diels-Alder reactions as the key step. Jacobsen chromium(III) catalyst and a titanium-binaphthol complex have been used to successfully catalyze diene and aldehyde cycloadditions, leading to optically active dihydropyran templates. 6-Hydroxydesosamine, orthogonally protected ezoaminuroic acid, and neosidomycin were synthesized using a comparative study. Also, a novel chiron approach to 4-deoxy-lyxo-hexopyranosiduronic acid methyl ester derivatives was efficiently accomplished starting from readily accessible starting materials. This work represents a systematic and comprehensive study toward a de novo synthesis of 4-deoxy-hexopyranoses via enantioselective hetero-Diels-Alder reactions.
Asunto(s)
Desoxiglucosa/análogos & derivados , Desoxiglucosa/síntesis química , Indoles/síntesis química , Ciclización , Desoxiglucosa/química , Indoles/química , Estructura Molecular , EstereoisomerismoRESUMEN
2-NBDG is a widely used fluorescent tracer for monitoring d-glucose uptake into single living cells. However, 2-NBDG alone is not sufficient for monitoring the net stereoselective uptake of d-glucose, unless its possible non-stereoselective uptake is properly evaluated. l-Glucose derivatives, which emit fluorescence distinct from that of 2-NBDG, should provide valuable information on the stereoselective uptake, when used with 2-NBDG in combination. In the present study, we synthesized Texas Red (sulforhodamine 101 acid)-coupled and [2-(benz-2-oxa-1,3-diazol-4-yl)amino]-coupled 2-deoxy-D-glucose, referred to as [2-TRG] and [2-BDG], respectively. These derivatives showed emission wavelength longer and shorter than that of 2-NBDG, respectively. 2-TRLG, an antipode of 2-TRG, proved to be an effective tracer for evaluating the extent of non-stereoselective uptake of 2-NBDG when used simultaneously with 2-NBDG. On the other hand, 2-BDG exhibited very weak fluorescence, but the application of a novel cross coupling in the presence of a benzoxadiazole group may be useful for the future development of effective glucose tracers.
Asunto(s)
4-Cloro-7-nitrobenzofurazano/análogos & derivados , Desoxiglucosa/análogos & derivados , Glucosa/metabolismo , 4-Cloro-7-nitrobenzofurazano/síntesis química , 4-Cloro-7-nitrobenzofurazano/química , 4-Cloro-7-nitrobenzofurazano/metabolismo , Animales , Encéfalo/metabolismo , Desoxiglucosa/síntesis química , Desoxiglucosa/química , Desoxiglucosa/metabolismo , Glucosa/química , Ratones , Microscopía Confocal , Estructura Molecular , EstereoisomerismoRESUMEN
This work provides evidence of previously unrecognized uptake of glucose via sodium-coupled glucose transporters (SGLTs) in specific regions of the brain. The current understanding of functional glucose utilization in brain is largely based on studies using positron emission tomography (PET) with the glucose tracer 2-deoxy-2-[F-18]fluoro-D-glucose (2-FDG). However, 2-FDG is only a good substrate for facilitated-glucose transporters (GLUTs), not for SGLTs. Thus, glucose accumulation measured by 2-FDG omits the role of SGLTs. We designed and synthesized two high-affinity tracers: one, α-methyl-4-[F-18]fluoro-4-deoxy-D-glucopyranoside (Me-4FDG), is a highly specific SGLT substrate and not transported by GLUTs; the other one, 4-[F-18]fluoro-4-deoxy-D-glucose (4-FDG), is transported by both SGLTs and GLUTs and will pass through the blood brain barrier (BBB). In vitro Me-4FDG autoradiography was used to map the distribution of uptake by functional SGLTs in brain slices with a comparable result from in vitro 4-FDG autoradiography. Immunohistochemical assays showed that uptake was consistent with the distribution of SGLT protein. Ex vivo 4-FDG autoradiography showed that SGLTs in these areas are functionally active in the normal in vivo brain. The results establish that SGLTs are a normal part of the physiology of specific areas of the brain, including hippocampus, amygdala, hypothalamus, and cerebral cortices. 4-FDG PET imaging also established that this BBB-permeable SGLT tracer now offers a functional imaging approach in humans to assess regulation of SGLT activity in health and disease.
Asunto(s)
Encéfalo/metabolismo , Glucosa/metabolismo , Proteínas de Transporte de Sodio-Glucosa/metabolismo , Animales , Barrera Hematoencefálica/metabolismo , Desoxiglucosa/análogos & derivados , Desoxiglucosa/síntesis química , Desoxiglucosa/metabolismo , Femenino , Glucósidos/síntesis química , Glucósidos/metabolismo , Proteínas de Transporte de Glutamato en la Membrana Plasmática/metabolismo , Tomografía de Emisión de Positrones , Ratas , Ratas Sprague-Dawley , Proteínas de Transporte de Sodio-Glucosa/análisisRESUMEN
beta-N-Acetylhexosaminidases feature so-called wobbling specificity, which means that they cleave substrates both in gluco- and galacto- configurations, with the activity ratio depending on the enzyme source. Here we present the new finding that fungal beta-N-acetylhexosaminidases are able to hydrolyze and transfer 4-deoxy-N-acetylhexosaminides with high yields. This clearly demonstrates that the 4-hydroxy moiety at the substrate pyranose ring is not essential for substrate binding to the enzyme active site, which was also confirmed by molecular docking of the tested compounds into the model of the active site of beta-N-acetylhexosaminidase from Aspergillus oryzae. A set of four 4-deoxy-N-acetylhexosaminides was synthesized and screened against a panel of beta-N-acetylhexosaminidases (extracellular and intracellular) from various sources (fungal, human, animal, plant and bacterial) for hydrolysis. The results of this screening are reported here, as well as the structures of three novel 4'-deoxy-disaccharides prepared by transglycosylation reaction with high yields (52% total disaccharide fraction) using beta-N-acetylhexosaminidase from Talaromyces flavus.
Asunto(s)
Desoxiglucosa/análogos & derivados , Glucosamina/análogos & derivados , beta-N-Acetilhexosaminidasas/metabolismo , Desoxiglucosa/síntesis química , Desoxiglucosa/química , Glucosamina/síntesis química , Glucosamina/química , Estructura Molecular , Estereoisomerismo , Especificidad por Sustrato , beta-N-Acetilhexosaminidasas/químicaRESUMEN
The conversion of cyclopropane-fused carbohydrates into oxepines is an attractive method for accessing diverse members of the septanoside family of carbohydrate mimetics. 2-Bromooxepines are obtained through silver(I)-promoted thermal ring expansion of a d-glucal-derived gem-dihalocyclopropanated sugar. In contrast, cyclopropane ring cleavage under basic conditions leads to 2-C-branched pyranosides, not the 2-bromooxepine structures assigned in an earlier report.
Asunto(s)
Carbohidratos/química , Ciclopropanos/química , Desoxiglucosa/análogos & derivados , Compuestos Epoxi/síntesis química , Oxepinas/síntesis química , Desoxiglucosa/síntesis química , Desoxiglucosa/química , Compuestos Epoxi/química , Espectroscopía de Resonancia Magnética , Estructura Molecular , Oxepinas/química , EstereoisomerismoRESUMEN
The deoxyglucose dithiocarbamate (DGDTC) was synthesized and radiolabelled with [(99m)TcN](2+) intermediate to form the (99m)TcN-DGDTC complex. The radiochemical purity of the (99m)TcN-DGDTC complex was over 90%, as measured by TLC and by HPLC, without any notable decomposition at room temperature over a period of 6h. The partition coefficient and electrophoresis results indicated that this complex was hydrophilic and neutral. The biodistribution of (99m)TcN-DGDTC in mice bearing S 180 tumor showed that the complex accumulated in the tumor with high uptake and good retention. The tumor/blood and tumor/muscle ratios increased with time and reached 2.32 and 1.68 at 4h post-injection, suggesting it would be a promising candidate for tumor imaging.
Asunto(s)
Neoplasias/diagnóstico por imagen , Compuestos de Organotecnecio/síntesis química , Radiofármacos/síntesis química , Animales , Carbamatos/síntesis química , Carbamatos/química , Desoxiglucosa/análogos & derivados , Desoxiglucosa/síntesis química , Desoxiglucosa/química , Marcaje Isotópico , Ratones , Compuestos de Organotecnecio/química , Cintigrafía , Radiofármacos/química , Distribución TisularRESUMEN
A straightforward synthesis of a derivative of anthrose, the non-reducing terminal fragment of the antigenic tetrasaccharide from Bacillus anthracis, was achieved starting from d-galactose. This hapten is able to induce a highly specific and sensitive immune response in rabbit when attached to a carrier protein.
Asunto(s)
Amino Azúcares/síntesis química , Bacillus anthracis/inmunología , Desoxiglucosa/análogos & derivados , Esporas Bacterianas/inmunología , Animales , Anticuerpos Antibacterianos/inmunología , Desoxiglucosa/síntesis química , Hemocianinas/inmunología , Técnicas para Inmunoenzimas/métodos , Conejos , Esporas Bacterianas/químicaRESUMEN
This protocol details a method for monitoring glucose uptake into single, living mammalian cells using a fluorescent D-glucose derivative, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG), as a tracer. The specifically designed chamber and superfusion system for evaluating 2-NBDG uptake into cells in real time can be combined with other fluorescent methods such as Ca2+ imaging and the subsequent immunofluorescent classification of cells exhibiting divergent 2-NBDG uptake. The whole protocol, including immunocytochemistry, can be completed within 2 d (except for cell culture). The procedure for 2-NBDG synthesis is also presented.