(2R,3R,4S,5S,6R)-2-amino-6-(hydroxymethyl)oxane-3,4,5-triol

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 1.79 g (10 mmol) of 1-amino-1-deoxy-β-D-glucopyranose is dissolved in a mixture of 20 ml methanol and 10 ml dioxane, and to this solution is added a mixture of formic acid and citric acid dropwise, with stirring, to adjust the PH of the solution to 9.1. The resulting solution is then added dropwise to a solution prepared by dissolving 3.28 g (12 mmol) of o-nitrophenyl N-(2-chloroethyl)-N-nitrosocarbamate in a mixture of 10 ml tetrahydrofuran and 15 ml dioxane, while stirring, at 5°-10° C. over 30 minutes. The resultant mixture is stirred at the same temperature for a further 4 hours, and the thus reacted solution is concentrated at 40° C. and below, under reduced pressure. The crystalline residue obtained is washed with anhydrous ethyl ether and then with acetone, dried under reduced pressure, and recrystallized from anhydrous ethanol-ethyl ether (4:1) to give 2.56 g of 3-(β-D-glucopyranosyl)-1-(2-chloroethyl)-1-nitrosourea. Yield 81.5%. mp 77°-78° C. (decomp.). [α]D25 -10.3° (C 1.0, water). Analysis for C9H16N3O7Cl (MW 313.69): to this solution is added The resulting solution is then added dropwise to a solution prepared while stirring, at 5°-10° C. over 30 minutes the thus reacted solution is concentrated at 40° C. The crystalline residue obtained is washed with anhydrous ethyl ether with acetone, dried under reduced pressure recrystallized from anhydrous ethanol-ethyl ether (4:1)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 1.79 g (10 mmol) of 1-amino-1-deoxy-β-D-glucopyranose is dissolved in a mixture of 20 ml methanol and 10 ml dioxane, and to this solution is added a mixture of formic acid and citric acid dropwise, with stirring, to adjust the PH of the solution to 9.1. The resulting solution is then added dropwise to a solution prepared by dissolving 3.28 g (12 mmol) of o-nitrophenyl N-(2-chloroethyl)-N-nitrosocarbamate in a mixture of 10 ml tetrahydrofuran and 15 ml dioxane, while stirring, at 5°-10° C. over 30 minutes. The resultant mixture is stirred at the same temperature for a further 4 hours, and the thus reacted solution is concentrated at 40° C. and below, under reduced pressure. The crystalline residue obtained is washed with anhydrous ethyl ether and then with acetone, dried under reduced pressure, and recrystallized from anhydrous ethanol-ethyl ether (4:1) to give 2.56 g of 3-(β-D-glucopyranosyl)-1-(2-chloroethyl)-1-nitrosourea. Yield 81.5%. mp 77°-78° C. (decomp.). [α]D25 -10.3° (C 1.0, water). Analysis for C9H16N3O7Cl (MW 313.69): to this solution is added The resulting solution is then added dropwise to a solution prepared while stirring, at 5°-10° C. over 30 minutes the thus reacted solution is concentrated at 40° C. The crystalline residue obtained is washed with anhydrous ethyl ether with acetone, dried under reduced pressure recrystallized from anhydrous ethanol-ethyl ether (4:1)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 1.79 g (10 mmol) of 1-amino-1-deoxy-β-D-glucopyranose is dissolved (partially suspended) in 40 ml of anhydrous ethanol, and the solution is cooled to 0°-5° C. To this solution is added 2.2 g (50 mmol) of dry-ice little by little so as to convert the glucopyranose into its carbonic acid-addition salt. This acid-addition salt solution is then added dropwise to a solution prepared by dissolving 3.28 g (12 mmol) of o-nitrophenyl N-(2-chloroethyl)-N-nitrosocarbamate in a mixture of 40 ml anhydrous tetrahydrofuran and 10 ml benzene, while stirring, at 0°-5° C. over an hour. The resultant mixture is stirred further at 20°-25° C. for 2 hours, and the thus reacted solution is concentrated at 50° C. and below, under reduced pressure. The crystalline residue obtained is washed with anhydrous ethyl ether and then with acetone, dried under reduced pressure, and recrystallized from anhydrous isopropanol to give 2.72 g of 3-(β-D-glucopyranosyl)-1-(2 -chloroethyl)-1-nitrosourea. Yield 86.8%. mp 77°-78° C. (decomp.). [α]D25 -10.3° (C 1.0, water). Analysis for C9H16N3O7Cl (MW 313.69): the solution is cooled to 0°-5° C This acid-addition salt solution is then added dropwise to a solution prepared the thus reacted solution is concentrated at 50° C. The crystalline residue obtained is washed with anhydrous ethyl ether with acetone, dried under reduced pressure recrystallized from anhydrous isopropanol

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 3.28 g (12 mmol) of o-nitrophenyl N-(2-chloroethyl)-N-nitrosocarbamate is dissolved in a mixture of 10 ml anhydrous dimethylformamide and 10 ml anhydrous ethanol, and to this solution is added little by little 1.79 g (10 mmol) of 1-amino-1-deoxy-β-D-glucopyranose powder, while stirring, at 5°-10° C. over 10 minutes. The resultant mixture is, after adding 5 ml of anhydrous dimethylformamide thereto, stirred further at 10° C. for 5 hours, and the thus reacted solution is concentrated at 40° C. and below, under reduced pressure. The crystalline residue obtained is washed with anhydrous ethyl ether and then with acetone, dried under reduced pressure, and recrystallized from anhydrous ethanol-ethyl ether (1:5) to give 2.47 g of 3-(β-D-glucopyranosyl)-1-(2-chloroethyl)-1-nitrosourea. Yield 78.7%. mp 77°-78° C. (decomp.). [α]D25 -10.3° (C 1.0, water). Analysis for C9H16N3O7Cl (MW 313.69): stirred further at 10° C. for 5 hours the thus reacted solution is concentrated at 40° C. The crystalline residue obtained is washed with anhydrous ethyl ether with acetone, dried under reduced pressure recrystallized from anhydrous ethanol-ethyl ether (1:5)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 1.79 g (10 mmol) of 1-amino-1-deoxy-β-D-glucopyranose is dissolved in a mixture of 20 ml methanol and 10 ml dioxane, and to this solution is added a mixture of formic acid and citric acid dropwise, with stirring, to adjust the PH of the solution to 9.1. The resulting solution is then added dropwise to a solution prepared by dissolving 3.28 g (12 mmol) of o-nitrophenyl N-(2-chloroethyl)-N-nitrosocarbamate in a mixture of 10 ml tetrahydrofuran and 15 ml dioxane, while stirring, at 5°-10° C. over 30 minutes. The resultant mixture is stirred at the same temperature for a further 4 hours, and the thus reacted solution is concentrated at 40° C. and below, under reduced pressure. The crystalline residue obtained is washed with anhydrous ethyl ether and then with acetone, dried under reduced pressure, and recrystallized from anhydrous ethanol-ethyl ether (4:1) to give 2.56 g of 3-(β-D-glucopyranosyl)-1-(2-chloroethyl)-1-nitrosourea. Yield 81.5%. mp 77°-78° C. (decomp.). [α]D25 -10.3° (C 1.0, water). Analysis for C9H16N3O7Cl (MW 313.69): to this solution is added The resulting solution is then added dropwise to a solution prepared while stirring, at 5°-10° C. over 30 minutes the thus reacted solution is concentrated at 40° C. The crystalline residue obtained is washed with anhydrous ethyl ether with acetone, dried under reduced pressure recrystallized from anhydrous ethanol-ethyl ether (4:1)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 1.79 g (10 mmol) of 1-amino-1-deoxy-β-D-glucopyranose is dissolved (partially suspended) in 40 ml of anhydrous ethanol, and the solution is cooled to 0°-5° C. To this solution is added 2.2 g (50 mmol) of dry-ice little by little so as to convert the glucopyranose into its carbonic acid-addition salt. This acid-addition salt solution is then added dropwise to a solution prepared by dissolving 3.28 g (12 mmol) of o-nitrophenyl N-(2-chloroethyl)-N-nitrosocarbamate in a mixture of 40 ml anhydrous tetrahydrofuran and 10 ml benzene, while stirring, at 0°-5° C. over an hour. The resultant mixture is stirred further at 20°-25° C. for 2 hours, and the thus reacted solution is concentrated at 50° C. and below, under reduced pressure. The crystalline residue obtained is washed with anhydrous ethyl ether and then with acetone, dried under reduced pressure, and recrystallized from anhydrous isopropanol to give 2.72 g of 3-(β-D-glucopyranosyl)-1-(2 -chloroethyl)-1-nitrosourea. Yield 86.8%. mp 77°-78° C. (decomp.). [α]D25 -10.3° (C 1.0, water). Analysis for C9H16N3O7Cl (MW 313.69): the solution is cooled to 0°-5° C This acid-addition salt solution is then added dropwise to a solution prepared the thus reacted solution is concentrated at 50° C. The crystalline residue obtained is washed with anhydrous ethyl ether with acetone, dried under reduced pressure recrystallized from anhydrous isopropanol

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 3.28 g (12 mmol) of o-nitrophenyl N-(2-chloroethyl)-N-nitrosocarbamate is dissolved in a mixture of 10 ml anhydrous dimethylformamide and 10 ml anhydrous ethanol, and to this solution is added little by little 1.79 g (10 mmol) of 1-amino-1-deoxy-β-D-glucopyranose powder, while stirring, at 5°-10° C. over 10 minutes. The resultant mixture is, after adding 5 ml of anhydrous dimethylformamide thereto, stirred further at 10° C. for 5 hours, and the thus reacted solution is concentrated at 40° C. and below, under reduced pressure. The crystalline residue obtained is washed with anhydrous ethyl ether and then with acetone, dried under reduced pressure, and recrystallized from anhydrous ethanol-ethyl ether (1:5) to give 2.47 g of 3-(β-D-glucopyranosyl)-1-(2-chloroethyl)-1-nitrosourea. Yield 78.7%. mp 77°-78° C. (decomp.). [α]D25 -10.3° (C 1.0, water). Analysis for C9H16N3O7Cl (MW 313.69): stirred further at 10° C. for 5 hours the thus reacted solution is concentrated at 40° C. The crystalline residue obtained is washed with anhydrous ethyl ether with acetone, dried under reduced pressure recrystallized from anhydrous ethanol-ethyl ether (1:5)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Beta-D-glucosylamine is prepared by the reaction of D-(+)-glucose with ammonium bicarbonate as described by Likhosherstov, L. M. et al. (Carbohydr. Res. 146: C1, 1986). A reaction mixture containing the glycosylamine and a 1- to 2-fold excess by weight of 5-aminosalicylic acid is allowed to react overnight in pyridine at 50° C. The reaction is monitored by thin-layer chromatography (TLC) (silica gel; eluent: CHCl3:MeOH, 1:1, by volume). The solvent is removed, and the reaction mixture is purified by chromatography on silica gel or BIOGEL-F2™ (BioRad, Richmond, Calif.), using absorbance at 254 nm as detection. The products are characterized by TLC on silica gel, electrospray mass spectrometry, and 1H- and 13C-NMR analysis. The transamination reaction is indicated by a downfield shift of resonance of the anomeric proton (observed at δ=4.1 ppm by analysis of the unsubstituted glycosyl-1-amine) to δ=4.3–4.7 ppm in the 1H-NMR spectrum of the β-anomer of the synthetic glycoconjugate product. (This anomer is the major product of the transamination reaction.) The solvent is removed the reaction mixture is purified by chromatography on silica gel or BIOGEL-F2™ (BioRad, Richmond, Calif.) The transamination reaction the major product of the transamination reaction

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 1.79 g (10 mmol) of 1-amino-1-deoxy-β-D-glucopyranose is dissolved in a mixture of 20 ml methanol and 10 ml dioxane, and to this solution is added a mixture of formic acid and citric acid dropwise, with stirring, to adjust the PH of the solution to 9.1. The resulting solution is then added dropwise to a solution prepared by dissolving 3.28 g (12 mmol) of o-nitrophenyl N-(2-chloroethyl)-N-nitrosocarbamate in a mixture of 10 ml tetrahydrofuran and 15 ml dioxane, while stirring, at 5°-10° C. over 30 minutes. The resultant mixture is stirred at the same temperature for a further 4 hours, and the thus reacted solution is concentrated at 40° C. and below, under reduced pressure. The crystalline residue obtained is washed with anhydrous ethyl ether and then with acetone, dried under reduced pressure, and recrystallized from anhydrous ethanol-ethyl ether (4:1) to give 2.56 g of 3-(β-D-glucopyranosyl)-1-(2-chloroethyl)-1-nitrosourea. Yield 81.5%. mp 77°-78° C. (decomp.). [α]D25 -10.3° (C 1.0, water). Analysis for C9H16N3O7Cl (MW 313.69): to this solution is added The resulting solution is then added dropwise to a solution prepared while stirring, at 5°-10° C. over 30 minutes the thus reacted solution is concentrated at 40° C. The crystalline residue obtained is washed with anhydrous ethyl ether with acetone, dried under reduced pressure recrystallized from anhydrous ethanol-ethyl ether (4:1)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 1.79 g (10 mmol) of 1-amino-1-deoxy-β-D-glucopyranose is dissolved (partially suspended) in 40 ml of anhydrous ethanol, and the solution is cooled to 0°-5° C. To this solution is added 2.2 g (50 mmol) of dry-ice little by little so as to convert the glucopyranose into its carbonic acid-addition salt. This acid-addition salt solution is then added dropwise to a solution prepared by dissolving 3.28 g (12 mmol) of o-nitrophenyl N-(2-chloroethyl)-N-nitrosocarbamate in a mixture of 40 ml anhydrous tetrahydrofuran and 10 ml benzene, while stirring, at 0°-5° C. over an hour. The resultant mixture is stirred further at 20°-25° C. for 2 hours, and the thus reacted solution is concentrated at 50° C. and below, under reduced pressure. The crystalline residue obtained is washed with anhydrous ethyl ether and then with acetone, dried under reduced pressure, and recrystallized from anhydrous isopropanol to give 2.72 g of 3-(β-D-glucopyranosyl)-1-(2 -chloroethyl)-1-nitrosourea. Yield 86.8%. mp 77°-78° C. (decomp.). [α]D25 -10.3° (C 1.0, water). Analysis for C9H16N3O7Cl (MW 313.69): the solution is cooled to 0°-5° C This acid-addition salt solution is then added dropwise to a solution prepared the thus reacted solution is concentrated at 50° C. The crystalline residue obtained is washed with anhydrous ethyl ether with acetone, dried under reduced pressure recrystallized from anhydrous isopropanol

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 3.28 g (12 mmol) of o-nitrophenyl N-(2-chloroethyl)-N-nitrosocarbamate is dissolved in a mixture of 10 ml anhydrous dimethylformamide and 10 ml anhydrous ethanol, and to this solution is added little by little 1.79 g (10 mmol) of 1-amino-1-deoxy-β-D-glucopyranose powder, while stirring, at 5°-10° C. over 10 minutes. The resultant mixture is, after adding 5 ml of anhydrous dimethylformamide thereto, stirred further at 10° C. for 5 hours, and the thus reacted solution is concentrated at 40° C. and below, under reduced pressure. The crystalline residue obtained is washed with anhydrous ethyl ether and then with acetone, dried under reduced pressure, and recrystallized from anhydrous ethanol-ethyl ether (1:5) to give 2.47 g of 3-(β-D-glucopyranosyl)-1-(2-chloroethyl)-1-nitrosourea. Yield 78.7%. mp 77°-78° C. (decomp.). [α]D25 -10.3° (C 1.0, water). Analysis for C9H16N3O7Cl (MW 313.69): stirred further at 10° C. for 5 hours the thus reacted solution is concentrated at 40° C. The crystalline residue obtained is washed with anhydrous ethyl ether with acetone, dried under reduced pressure recrystallized from anhydrous ethanol-ethyl ether (1:5)