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

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

Procedure: 1.79 g (10 mmol) of 6-amino-6-deoxy-D-glucopyranose is dissolved in a mixture of 20 ml anhydrous ethanol and 20 ml dimethyl sulfoxide, and the solution is cooled to 0°-5° C. To this solution is added 3.3 g (75 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 toluene, while stirring, at 0°-5° C. over 30 minutes. The resultant mixture is stirred further at 20°-25° C. for 2 hours, and the thus reacted solution is concentrated at 40° C. and below, under reduced pressure. To the oily residue thus produced is added 80 ml of ethyl ether and the solution is left at 4° C. overnight. The crystalline residue obtained is washed with chloroform and then with ethyl ether, dried under reduced pressure, and recrystallized from anhydrous ethanol-n-hexane (4:1) to give 2.70 g of 3-(D-glucopyranos-6-yl)-1-(2-chloroethyl)-1-nitrosourea. Yield 86.1%. mp 102°-103° C. (decomp.). [α]D20 +55° (C 0.5, methanol; the value of +55° reduced to +37° after 18 hours). 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 40° C. To the oily residue thus produced the solution is left at 4° C. overnight The crystalline residue obtained is washed with chloroform with ethyl ether, dried under reduced pressure recrystallized from anhydrous ethanol-n-hexane (4:1)

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

Procedure: 1.79 g (10 mmol) of 2-amino-2-deoxy-D-glucopyranose is dissolved in a mixture of 20 ml anhydrous methanol and 10 ml dimethylformamide, 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 said glucopyranose into its carbonic acid-addition salt. This acid-addition salt solution is then added dropwise to a solution prepared by dissolving 3.04 g (12 mmol) of o-cyanophenyl N-(2-chloroethyl)-N-nitrosocarbamate in a mixture of 20 ml anhydrous tetrahydrofuran and 10 ml methylene chloride, while stirring, at 0°-5° C. over an hour. The resultant mixture is stirred further at 22°-25° C. for 2 hours, and the thus reacted solution is concentrated at 40° C. and below, under reduced pressure. The crystalline residue obtained is washed with petroleum ether, dried under reduced pressure, and recrystallized from anhydrous ethanol to give 2.58 g of 3-(D-glucopyranos-2-yl)-1-(2-chloroethyl)-1-nitrosourea. Yield 82.2%. mp 147°-148° C. (decomp.). 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 40° C. The crystalline residue obtained is washed with petroleum ether dried under reduced pressure recrystallized from anhydrous ethanol

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: 1.79 g (10 mmol) of 2-amino-2-deoxy-D-glucopyranose is dissolved in a mixture of 20 ml anhydrous methanol and 10 ml dimethylformamide, 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 said glucopyranose into its carbonic acid-addition salt. This acid-addition salt solution is then added dropwise to a solution prepared by dissolving 3.04 g (12 mmol) of o-cyanophenyl N-(2-chloroethyl)-N-nitrosocarbamate in a mixture of 20 ml anhydrous tetrahydrofuran and 10 ml methylene chloride, while stirring, at 0°-5° C. over an hour. The resultant mixture is stirred further at 22°-25° C. for 2 hours, and the thus reacted solution is concentrated at 40° C. and below, under reduced pressure. The crystalline residue obtained is washed with petroleum ether, dried under reduced pressure, and recrystallized from anhydrous ethanol to give 2.58 g of 3-(D-glucopyranos-2-yl)-1-(2-chloroethyl)-1-nitrosourea. Yield 82.2%. mp 147°-148° C. (decomp.). 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 40° C. The crystalline residue obtained is washed with petroleum ether dried under reduced pressure recrystallized from anhydrous ethanol

Conditions: temp:35.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: D(+)-glucose monoydrate (300 g) and magnesium chloride hexahydrate (1.0 g) were dissolved in 10 mM MES buffer pH 6.5 (2.4 L; Sigma M3671). After addition of 4-oxo-6-aza-spiro[2.5]octane-6-carboxylic acid tert-butyl ester (300 g; 1.33 mmol) and □-NAD (3.0 g; free acid; Roche Diagnostics Cat. No. 10 004 626) the pH was re-adjusted and the suspension heated to 35° C. The reaction was started by adding ketoreductase KRED-NADH-117 (3.0 g; former Biocatalytics, now Codexis) and glucose dehydrogenase GDH-102 (300 mg; Biocatalytics). The suspension was vigorously stirred at 35° C. keeping the pH constant at 6.5 by the controlled addition (pH-stat) of 1.0 M aq. sodium hydroxide solution. After a consumption of 1.307 L (corresponding to 98% conversion; after 17 h) the reaction mixture was extracted with EtOAc (10 L). The organic phase was dried over sodium sulfate and concentrated in vacuo (200 mbar/45° C.) until evaporation fell off. Upon cooling the oily residue (411 g) started to crystallize and was stirred with heptane (1 L) for 2 h. The crystals were filtered off and the filtrate evaporated to dryness, redissolved in EtOAc (150 ml) and concentrated in vacuo as described above. The crystal suspension formed again upon cooling was stirred with heptane (200 ml; 2 h) and the crystals filtered off. Both crops of crystals were washed with heptane and dried under high vacuum to yield the title compound in 93% yield (250.77 g and 34.60 g white crystals), each having a purity of >98.5% GC and 99.8% ee. [α]D=−44.97° (c=1.00, CHCl3). After a consumption of 1.307 L (corresponding to 98% conversion; after 17 h) the reaction mixture was extracted with EtOAc (10 L) The organic phase was dried over sodium sulfate concentrated in vacuo (200 mbar/45° C.) until evaporation Upon cooling the oily residue (411 g) to crystallize was stirred with heptane (1 L) for 2 h The crystals were filtered off the filtrate evaporated to dryness redissolved in EtOAc (150 ml) concentrated in vacuo The crystal suspension formed again upon cooling the crystals filtered off Both crops of crystals were washed with heptane dried under high vacuum

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

Procedure: To 11.33 g. (0.10 mole) of methyl acetimidate hydrochloride, prepared by known procedures, in a 100 ml. 3-necked, round-bottomed flask there was added a solution of 9.71 g. (actual 9.42 g.; 49.6 mmole) of crude p-isobutylpropiophenone, prepared as described in Part A above, in 23 ml. of absolute methanol. The resulting solution was stirred for 12 hours at room temperature to insure complete reaction. Gas liquid chromatographic analysis (Glc analysis) of an aliquot of the reaction mixture indicated greater than 99% ketal formation. The resulting mixture was filtered to remove the precipitated ammonium chloride and concentrated under vacuum. Hexane (50 ml.) was added to the residue and the resulting solution was again filtered to remove any acetamide which might be present. Removal of the hexane solvent under vacuum gave p-isobutylpropiophenone dimethyl ketal as a pale yellow oil which was used without further purification. The NMR was in accord. 3-necked, round-bottomed flask there was added a solution of 9.71 g reaction The resulting mixture was filtered to remove the precipitated ammonium chloride concentrated under vacuum Hexane (50 ml.) was added to the residue the resulting solution was again filtered to remove any acetamide which Removal of the hexane solvent under vacuum

Conditions: temp:35.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: D(+)-glucose monoydrate (300 g) and magnesium chloride hexahydrate (1.0 g) were dissolved in 10 mM MES buffer pH 6.5 (2.4 L; Sigma M3671). After addition of 4-oxo-6-aza-spiro[2.5]octane-6-carboxylic acid tert-butyl ester (300 g; 1.33 mmol) and β-NAD (3.0 g; free acid; Roche Diagnostics Cat. No. 10 004 626) the pH was re-adjusted and the suspension heated to 35° C. The reaction was started by adding ketoreductase KRED-NADH-117 (3.0 g; former Biocatalytics, now Codexis) and glucose dehydrogenase GDH-102 (300 mg; Biocatalytics). The suspension was vigorously stirred at 35° C. keeping the pH constant at 6.5 by the controlled addition (pH-stat) of 1.0 M aq. sodium hydroxide solution. After a consumption of 1.307 L (corresponding to 98% conversion; after 17 h) the reaction mixture was extracted with ethyl acetate (10 L). The organic phase was dried over sodium sulfate and concentrated in vacuo (200 mbar/45° C.) until evaporation fell off. Upon cooling the oily residue (411 g) started to crystallize and was stirred with heptane (1 L) for 2 h. The crystals were filtered off and the filtrate evaporated to dryness, redissolved in ethyl acetate (150 ml) and concentrated in vacuo as described above. The crystal suspension formed again upon cooling was stirred with heptane (200 ml; 2 h) and the crystals filtered off. Both crops of crystals were washed with heptane and dried under high vacuum to yield the title compound in 93% yield (250.77 g and 34.60 g white crystals), each having a purity of >98.5% GC and 99.8% ee. [α]D=−44.97° (c=1.00, CHCl3). After a consumption of 1.307 L (corresponding to 98% conversion; after 17 h) the reaction mixture was extracted with ethyl acetate (10 L) The organic phase was dried over sodium sulfate concentrated in vacuo (200 mbar/45° C.) until evaporation Upon cooling the oily residue (411 g) to crystallize was stirred with heptane (1 L) for 2 h The crystals were filtered off the filtrate evaporated to dryness redissolved in ethyl acetate (150 ml) concentrated in vacuo The crystal suspension formed again upon cooling the crystals filtered off Both crops of crystals were washed with heptane dried under high vacuum

Conditions: temp:35.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: D(+)-glucose monoydrate (300 g) and magnesium chloride hexahydrate (1.0 g) were dissolved in 10 mM MES buffer pH 6.5 (2.4 L; Sigma M3671). After addition of 4-oxo-6-aza-spiro[2.5]octane-6-carboxylic acid tert-butyl ester (300 g; 1.33 mmol) and β-NAD (3.0 g; free acid; Roche Diagnostics Cat. No. 10 004 626) the pH was re-adjusted and the suspension heated to 35° C. The reaction was started by adding ketoreductase KRED-NADH-117 (3.0 g; former Biocatalytics, now Codexis) and glucose dehydrogenase GDH-102 (300 mg; Biocatalytics). The suspension was vigorously stirred at 35° C. keeping the pH constant at 6.5 by the controlled addition (pH-stat) of 1.0 M aq. sodium hydroxide solution. After a consumption of 1.307 L (corresponding to 98% conversion; after 17 h) the reaction mixture was extracted with ethyl acetate (10 L). The organic phase was dried over sodium sulfate and concentrated in vacuo (200 mbar/45° C.) until evaporation fell off. Upon cooling the oily residue (411 g) started to crystallize and was stirred with heptane (1 L) for 2 h. The crystals were filtered off and the filtrate evaporated to dryness, redissolved in ethyl acetate (150 ml) and concentrated in vacuo as described above. The crystal suspension formed again upon cooling was stirred with heptane (200 ml; 2 h) and the crystals filtered off. Both crops of crystals were washed with heptane and dried under high vacuum to yield the title compound in 93% yield (250.77 g and 34.60 g white crystals), each having a purity of >98.5% GC and 99.8% ee. [α]D=−44.97° (C=1.00, CHCl3). After a consumption of 1.307 L (corresponding to 98% conversion; after 17 h) the reaction mixture was extracted with ethyl acetate (10 L) The organic phase was dried over sodium sulfate concentrated in vacuo (200 mbar/45° C.) until evaporation Upon cooling the oily residue (411 g) to crystallize was stirred with heptane (1 L) for 2 h The crystals were filtered off the filtrate evaporated to dryness redissolved in ethyl acetate (150 ml) concentrated in vacuo The crystal suspension formed again upon cooling the crystals filtered off Both crops of crystals were washed with heptane dried under high vacuum

Conditions: temp:35.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: D(+)-glucose monoydrate (300 g) and magnesium chloride hexahydrate (1.0 g) were dissolved in 10 mM MES buffer pH 6.5 (2.4 L; Sigma M3671). After addition of 4-oxo-6-aza-spiro[2.5]octane-6-carboxylic acid tert-butyl ester (300 g; 1.33 mmol) and β-NAD (3.0 g; free acid; Roche Diagnostics Cat. No. 10 004 626) the pH was re-adjusted and the suspension heated to 35° C. The reaction was started by adding ketoreductase KRED-NADH-117 (3.0 g; former Biocatalytics, now Codexis) and glucose dehydrogenase GDH-102 (300 mg; Biocatalytics). The suspension was vigorously stirred at 35° C. keeping the pH constant at 6.5 by the controlled addition (pH-stat) of 1.0 M aq. sodium hydroxide solution. After a consumption of 1.307 L (corresponding to 98% conversion; after 17 h) the reaction mixture was extracted with ethyl acetate (10 L). The organic phase was dried over sodium sulfate and concentrated in vacuo (200 mbar/45° C.) until evaporation fell off. Upon cooling the oily residue (411 g) started to crystallize and was stirred with heptane (1 L) for 2 h. The crystals were filtered off and the filtrate evaporated to dryness, redissolved in ethyl acetate (150 ml) and concentrated in vacuo as described above. The crystal suspension formed again upon cooling was stirred with heptane (200 ml; 2 h) and the crystals filtered off. Both crops of crystals were washed with heptane and dried under high vacuum to yield the title compound in 93% yield (250.77 g and 34.60 g white crystals), each having a purity of >98.5% GC and 99.8% ee. [α]D=−44.97° (C=1.00, CHCl3). After a consumption of 1.307 L (corresponding to 98% conversion; after 17 h) the reaction mixture was extracted with ethyl acetate (10 L) The organic phase was dried over sodium sulfate concentrated in vacuo (200 mbar/45° C.) until evaporation Upon cooling the oily residue (411 g) to crystallize was stirred with heptane (1 L) for 2 h The crystals were filtered off the filtrate evaporated to dryness redissolved in ethyl acetate (150 ml) concentrated in vacuo The crystal suspension formed again upon cooling the crystals filtered off Both crops of crystals were washed with heptane dried under high vacuum

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

Procedure: 1.79 g (10 mmol) of 6-amino-6-deoxy-D-glucopyranose is dissolved in a mixture of 20 ml anhydrous ethanol and 20 ml dimethyl sulfoxide, and the solution is cooled to 0°-5° C. To this solution is added 3.3 g (75 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 toluene, while stirring, at 0°-5° C. over 30 minutes. The resultant mixture is stirred further at 20°-25° C. for 2 hours, and the thus reacted solution is concentrated at 40° C. and below, under reduced pressure. To the oily residue thus produced is added 80 ml of ethyl ether and the solution is left at 4° C. overnight. The crystalline residue obtained is washed with chloroform and then with ethyl ether, dried under reduced pressure, and recrystallized from anhydrous ethanol-n-hexane (4:1) to give 2.70 g of 3-(D-glucopyranos-6-yl)-1-(2-chloroethyl)-1-nitrosourea. Yield 86.1%. mp 102°-103° C. (decomp.). [α]D20 +55° (C 0.5, methanol; the value of +55° reduced to +37° after 18 hours). 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 40° C. To the oily residue thus produced the solution is left at 4° C. overnight The crystalline residue obtained is washed with chloroform with ethyl ether, dried under reduced pressure recrystallized from anhydrous ethanol-n-hexane (4:1)

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

Procedure: 1.79 g (10 mmol) of 2-amino-2-deoxy-D-glucopyranose is dissolved in a mixture of 20 ml anhydrous methanol and 10 ml dimethylformamide, 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 said glucopyranose into its carbonic acid-addition salt. This acid-addition salt solution is then added dropwise to a solution prepared by dissolving 3.04 g (12 mmol) of o-cyanophenyl N-(2-chloroethyl)-N-nitrosocarbamate in a mixture of 20 ml anhydrous tetrahydrofuran and 10 ml methylene chloride, while stirring, at 0°-5° C. over an hour. The resultant mixture is stirred further at 22°-25° C. for 2 hours, and the thus reacted solution is concentrated at 40° C. and below, under reduced pressure. The crystalline residue obtained is washed with petroleum ether, dried under reduced pressure, and recrystallized from anhydrous ethanol to give 2.58 g of 3-(D-glucopyranos-2-yl)-1-(2-chloroethyl)-1-nitrosourea. Yield 82.2%. mp 147°-148° C. (decomp.). 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 40° C. The crystalline residue obtained is washed with petroleum ether dried under reduced pressure recrystallized from anhydrous ethanol

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