Synthesis and biologic evaluation of (11)c-methyl-d-glucoside, a tracer of the sodium-dependent glucose transporters.

UNLABELLED: This study aimed to synthesize and to evaluate the biologic characteristics of (11)C labeled methyl-D-glucoside, a nonmetabolizable tracer that is selectively transported by sodium-dependent glucose transporters (SGLTs). METHODS: (11)C-Methyl-D-glucoside was prepared by methylation of glucose with (11)C-methyl triflate and was obtained as a mixture of anomers that were separated with high-performance liquid chromatography. The biodistribution of both the D- and L-isomers was determined in mice, and the presence of metabolites in the blood was investigated. The intrarenal distribution of (11)C-methyl-D-glucoside in mouse kidneys was visualized using autoradiography. Transport of alpha-methyl-D-glucoside and beta-methyl-D-glucoside by the human sodium-D-glucose cotransporter hSGLT1 was characterized after expression of hSGLT1 in oocytes of Xenopus laevis. RESULTS: The developed preparation procedure provided (11)C-methyl-D-glucoside in a total synthesis time of 20 min and a yield of 30% (decay corrected). The alpha- and beta-anomers of methyl-D-glucoside were reabsorbed from the primary urinary filtrate and showed only a minimal urinary excretion. Because methyl-L-glucoside was not reabsorbed and the reabsorption of methyl-D-glucoside was blocked by phlorizin, sodium-D-glucose cotransporters were critically involved. beta-Methyl-D-glucoside was accumulated in the kidneys to a higher extent than the alpha-anomer, suggesting that the basolateral efflux from the tubular cells is slower for the beta-anomer. Autoradiography showed that methyl-D-glucoside was accumulated throughout the renal cortex, suggesting that both sodium-D-glucose cotransporters expressed in kidney, SGLT1 and SGLT2, are involved in the uptake. The tracer was found to be metabolically stable and did not accumulate in red blood cells, which indicates that methyl-D-glucoside is not transported by the sodium-independent transporter GLUT1. Electrical measurements in Xenopus oocytes revealed that alpha-methyl-D-glucoside and beta-methyl-D-glucoside are transported by the human SGLT1 transporter with similar maximal transport rates and apparent Michaelis-Menten constant values. CONCLUSION: (11)C-Methyl-D-glucoside is a selective tracer of sodium-dependent glucose transport and can be used to visualize the function of this transporter with PET in vivo.

A novel HPLC method for the simultaneous quantification of monosaccharides and disaccharides used in tests of intestinal function and permeability.

The quantification of monosaccharides and disaccharides used as probes in intestinal function and permeability tests can be technically demanding, detracting from the value of this approach to the indirect assessment of intestinal damage. In this study, a procedure is described for the simultaneous quantification of rhamnose, lactulose, 3-O-methyl-D-glucose and xylose in urine by HPLC using an anion exchange column with pulsed amperometric detection. This method is relatively fast and simple to perform, requiring no pre-treatment of urine samples or post-column derivatization. Accuracy and precision of determinations are illustrated by analytical recoveries (mean percentage +/- S.D., CV., n = 30) for multiple batch analyses of a diluted urine sample containing 20 mg/l of rhamnose (100 +/- 6.8, 6.2%), lactulose (100 +/- 6.1, 5.5%), 3-O-methyl-D-glucose (98 +/- 5.9, 5.5%) and xylose (104 +/- 7.1, 6.5%). Linearity of standard curves indicated that the lower limit for accurate quantification was 0.1 mg/l for all four sugars. Urinary recoveries following oral administration of these sugars to control dogs were determined as a baseline for the investigation of intestinal damage in this species and comparison of chromatograms illustrated enhanced permeability in dogs with gluten-sensitive enteropathy.

The separation of methyl ethers of methyl 2-acetamido-2-deoxy-D-glucopyranosides on a strongly basic ion-exchange resin.

The methyl ethers of methyl 2-acetamido-2-deoxy-alpha- and -beta-D-glucopyranoside can be isolated on a preparative scale by chromatography on Dowex-1(HO-) resin. This procedure greatly simplifies the purification of methyl ethers, and has been used to isolate the methyl ethers produced by partial methylation of methyl 2-acetamido-2-deoxy-beta-D-glucopyranoside. The separations are thought to depend on an ion-exchange process in which all the free hydroxyl groups are involved. It is concluded that the following acidity sequence holds: HO-4 greater than HO-3 greater than HO-6.

Fast-atom-bombardment, negative-ion mass spectrometry of the mycobacterial O-methyl-D-glucose polysaccharide and lipopolysaccharides.

The mycobacterial O-methyl-D-glucose polysaccharide (MGP) and its acylated derivatives, the O-methyl-D-glucose lipopolysaccharides (MGLP), have been analyzed by fast-atom-bombardment mass spectrometry (F.a.b.m.s.). The molecular-ion peaks for MGP confirmed the degree of polymerization and, in addition, revealed a slight heterogeneity in the degree of methylation. Sequence ions were observed that are consistent with all known features of MGP structure. Spectra of the MGLP isomers confirmed the general distribution of the neutral and acidic acyl groups, and provided additional information regarding the specific location of individual acyl groups. The spectra of some MGLP preparations revealed that they were contaminated by lysophosphatidylinositol dimannoside. The study further documents the utility of F.a.b.m.s. for characterization of relatively large and complex carbohydrate derivatives.

KS-505a, a novel inhibitor of bovine brain Ca2+ and calmodulin-dependent cyclic-nucleotide phosphodiesterase from Streptomyces argenteolus.

A novel compound, KS-505a was isolated from the culture broth of a strain identified as Streptomyces argenteolus A-2. The compound inhibited bovine brain Ca2+ and calmodulin-dependent cyclic-nucleotide phosphodiesterase with an IC50 value (the concentration causing 50% inhibition) of 0.065 microM. The compound around that concentration had little or no effect on heart calmodulin-dependent and -independent cyclic-nucleotide phosphodiesterases, and protein kinase C.