methyl (3S)-4-bromo-3-hydroxybutanoate

反応条件: temp:92.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: A solution of (S)-4-Bromo-3-hydroxybutanoic acid, methyl ester (14.822 g, 75.2 mmol) in methyl ethyl ketone (255 mL) was treated with anhydrous sodium iodide (56.6 g, 376 mmol). The resulting red mixture was stirred for 23 hours under argon at a bath temperature of 92° C. The solution was allowed to cool to room temperature and the solids were filtered through celite. The receiver flask was switched and the celite bed was washed with ethyl acetate (3×50 mL). The original MEK product-rich filtrate was concentrated to approximately 60 mL. The residue was treated with the ethyl acetate wash from above followed by 250 mL of fresh ethyl acetate. The solution was washed with sodium sulfite solution (2×100 mL), brine (1×75 mL) and dried (sodium sulfate). Filtration followed by concentration in vacuo produced a yellow liquid which was evaporated several times from hexane and dried under high vacuum (5 mm) to 15.48 g (84%). to cool to room temperature the solids were filtered through celite the celite bed was washed with ethyl acetate (3×50 mL) The original MEK product-rich filtrate was concentrated to approximately 60 mL The residue was treated with the ethyl acetate wash The solution was washed with sodium sulfite solution (2×100 mL), brine (1×75 mL) dried (sodium sulfate) Filtration followed by concentration in vacuo produced a yellow liquid which was evaporated several times from hexane dried under high vacuum (5 mm) to 15.48 g (84%)

反応条件: temp:30.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: Methyl 4-bromo-3-oxobutyrate (300 mg), the above-mentioned wet bacterial cells (0.4 g), NADP+ (9 mg), glucose (750 mg), glucose dehydrogenase (manufactured by Amano Pharmaceuticals, Co., Ltd.) (1.2 mg), 100 mM phosphate buffer (pH 6.5) (15 ml) and butyl acetate (15 ml) are mixed. The mixture is stirred at 30° C. for 7 hr. During the stirring, 2M aqueous sodium carbonate solution is gradually added to adjust the pH of the reaction solution to 6.5±0.2. After the stirring has been finished, the reaction solution is centrifuged (1000×g, 5 min) to collect the organic layer. The organic layer is subjected to an analysis for content using gas chromatography under the following condition. The optical purity of methyl 4-bromo-3-hydroxybutyrate in the above-mentioned organic layer was also determined under the following condition. The organic layer was concentrated to give crude methyl (S)-4-bromo-3-hydroxybutyrate. (1000×g, 5 min) to collect the organic layer The organic layer was concentrated

反応条件: temp:30.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: Methyl 4-bromo-3-oxobutyrate (300 mg), the above-mentioned wet bacterial cells (0.4 g), NADP+ (9 mg), glucose (750 mg), glucose dehydrogenase (manufactured by Amano Pharmaceuticals, Co., Ltd.) (1.2 mg), 100 mM phosphate buffer (pH 6.5) (15 ml) and butyl acetate (15 ml) are mixed. The mixture is stirred at 30° C. for 7 hr. During the stirring, 2M aqueous sodium carbonate solution is gradually added to adjust the pH of the reaction solution to 6.5±0.2. After the stirring has been finished, the reaction solution is centrifuged (1000×g, 5 min) to collect the organic layer. The organic layer is subjected to an analysis for content using gas chromatography under the following condition. The optical purity of methyl 4-bromo-3-hydroxybutyrate in the above-mentioned organic layer was also determined under the following condition. The organic layer was concentrated to give crude methyl (S)-4-bromo-3-hydroxybutyrate. (1000×g, 5 min) to collect the organic layer The organic layer was concentrated

反応条件: temp:92.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: A solution of (S)-4-Bromo-3-hydroxybutanoic acid, methyl ester (14.822 g, 75.2 mmol) in methyl ethyl ketone (255 mL) was treated with anhydrous sodium iodide (56.6 g, 376 mmol). The resulting red mixture was stirred for 23 hours under argon at a bath temperature of 92° C. The solution was allowed to cool to room temperature and the solids were filtered through celite. The receiver flask was switched and the celite bed was washed with ethyl acetate (3×50 mL). The original MEK product-rich filtrate was concentrated to approximately 60 mL. The residue was treated with the ethyl acetate wash from above followed by 250 mL of fresh ethyl acetate. The solution was washed with sodium sulfite solution (2×100 mL), brine (1×75 mL) and dried (sodium sulfate). Filtration followed by concentration in vacuo produced a yellow liquid which was evaporated several times from hexane and dried under high vacuum (5 mm) to 15.48 g (84%). to cool to room temperature the solids were filtered through celite the celite bed was washed with ethyl acetate (3×50 mL) The original MEK product-rich filtrate was concentrated to approximately 60 mL The residue was treated with the ethyl acetate wash The solution was washed with sodium sulfite solution (2×100 mL), brine (1×75 mL) dried (sodium sulfate) Filtration followed by concentration in vacuo produced a yellow liquid which was evaporated several times from hexane dried under high vacuum (5 mm) to 15.48 g (84%)

反応条件: temp:92.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: A solution of (S)-4-Bromo-3-hydroxybutanoic acid, methyl ester (14.822 g, 75.2 mmol) in methyl ethyl ketone (255 mL) was treated with anhydrous sodium iodide (56.6 g, 376 mmol). The resulting red mixture was stirred for 23 hours under argon at a bath temperature of 92° C. The solution was allowed to cool to room temperature and the solids were filtered through celite. The receiver flask was switched and the celite bed was washed with ethyl acetate (3×50 mL). The original MEK product-rich filtrate was concentrated to approximately 60 mL. The residue was treated with the ethyl acetate wash from above followed by 250 mL of fresh ethyl acetate. The solution was washed with sodium sulfite solution (2×100 mL), brine (1×75 mL) and dried (sodium sulfate). Filtration followed by concentration in vacuo produced a yellow liquid which was evaporated several times from hexane and dried under high vacuum (5 mm) to 15.48 g (84%). to cool to room temperature the solids were filtered through celite the celite bed was washed with ethyl acetate (3×50 mL) The original MEK product-rich filtrate was concentrated to approximately 60 mL The residue was treated with the ethyl acetate wash The solution was washed with sodium sulfite solution (2×100 mL), brine (1×75 mL) dried (sodium sulfate) Filtration followed by concentration in vacuo produced a yellow liquid which was evaporated several times from hexane dried under high vacuum (5 mm) to 15.48 g (84%)

反応条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: A solution was prepared by dissolving 75 g of glucose, 0.85 g of oxidized form of nicotine adenine dinucleotide phosphate, 0.11 g of glucose dehydrogenase (glucose amino 2; by Amano enzyme) in 1500 g of a phosphate buffer solution (pH 6.5) and 1300 g of butyl acetate and 37 g of methyl 4-bromo-3-oxobutanoate were added thereto, and then 150 g of the acetone-treated cells of Penicillium citrinum IFO 463described above were added to thereto under stirring. 15% aqueous sodium carbonate solution was dropwise added thereto so that the pH of the reaction solution is maintained within a range of from 6.5±0.5. After stirring for 24 hours, 90 g of celite were added thereto and filtered under reduced pressure. The organic layer separated from the filtrate was concentrated under reduced pressure to give 28 g of methyl (S)-4-bromo-3-hydroxybutanoate, Chemical Purity: 88%, Optical purity: 97% e.e. A solution was prepared above were added to thereto so that the pH of the reaction solution is maintained within a range of from 6.5±0.5 After stirring for 24 hours 90 g of celite were added filtered under reduced pressure The organic layer separated from the filtrate was concentrated under reduced pressure

反応条件: temp:140.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: A mixture of (E)-3-[2-(Tributylstannyl)-ethenyl]-4-(4-fluoro-2-methylphenyl)-2-(1-methylethyl)-6-phenylpyridine (7.18 gm, 24.7 mmol), acetylene 1 (4.500 gm, 13.7 mmol) and AIBN (0.084 gm, 5.1 mmol) were heated at 140° C. for 1.5 hours. The solution was cooled to room temperature and diluted with ether (30 ml). Iodine (7.16 gm, 27.1 mmol) was added and an unexpectedly strong exothermic reaction occurred and approximately half the solution foamed out of the flask. The flask was recapped and allowed to stir 16 hours. The solution was then quenched with 10% Na2SO2O3 in saturated NaHCO3. The solution was diluted with ether, washed with 10% Na2SO2O3 in saturated NaHCO3 (twice) and brine, then dried over Na2SO4, filtered and concentrated to a yellow oil. The oil was purified by flash chromatography on Merck silica gel in 1% EtOAc in hexane. The desired fractions were combined and concentrated to afford a white solid, which was recrystallized from hexane to give product A as hard, off-white crystals (2.830 gm, 45%). The solution was cooled to room temperature an unexpectedly strong exothermic reaction The solution was then quenched with 10% Na2SO2O3 in saturated NaHCO3 The solution was diluted with ether washed with 10% Na2SO2O3 in saturated NaHCO3 (twice) and brine dried over Na2SO4 filtered concentrated to a yellow oil The oil was purified by flash chromatography on Merck silica gel in 1% EtOAc in hexane concentrated to afford a white solid, which was recrystallized from hexane

反応条件: temp:140.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: A mixture of (E)-3-[2-(Tributylstannyl)ethenyl]-4-(4-fluoro-2-methylphenyl)-2-(1-methylethyl)-6-phenylpyridine (7.18 gm, 24.7 mmol), acetylene 1 (4.500 gm, 13.7 mmol) and AIBN (0.084 gm, 5.1 mmol) were heated at 140° C. for 1.5 hours. The solution was cooled to room temperature and diluted with ether (30 ml). Iodine (7.16 gm, 27.1 mmol) was added and an unexpectedly strong exothermic reaction occurred and approximately half the solution foamed out of the flask. The flask was recapped and allowed to stir 16 hours. The solution was then quenched with 10% Na2S2O3 in saturated NaHCO3. The solution was diluted with ether, washed with 10% Na2S2O3 in saturated NaHCO3 (twice) and brine, then dried over Na2SO4, filtered and concentrated to a yellow oil. The oil was purified by flash chromatography on Merck silica gel in 1% EtOAc in hexane. The desired fractions were combined and concentrated to afford a white solid, which was recrystallized from hexane to give product A as hard, off-white crystals (2.830 gm, 45%). The solution was cooled to room temperature an unexpectedly strong exothermic reaction The solution was then quenched with 10% Na2S2O3 in saturated NaHCO3 The solution was diluted with ether washed with 10% Na2S2O3 in saturated NaHCO3 (twice) and brine dried over Na2SO4 filtered concentrated to a yellow oil The oil was purified by flash chromatography on Merck silica gel in 1% EtOAc in hexane concentrated to afford a white solid, which was recrystallized from hexane

反応条件: temp:140.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: A mixture of (E)-3-[2-(Tributylstannyl)ethenyl]-4-(4-fluoro-2-methylphenyl)-2-(1-methylethyl)-6-phenylpyridine (7.18 gm, 24.7 mmol), acetylene 1 (4.500 gm, 13.7 mmol) and AIBN (0.084 gm, 5.1 mmol) were heated at 140° C. for 1.5 hours. The solution was cooled to room temperature and diluted with ether (30 ml). Iodine (7.16 gm, 27.1 mmol) was added and an unexpectedly strong exothermic reaction occurred and approximately half the solution foamed out of the flask. The flask was recapped and allowed to stir 16 hours. The solution was then quenched with 10% Na2S2O3 in saturated NaHCO3. The solution was diluted with ether, washed with 10% Na2S2O3 in saturated NaHCO3 (twice) and brine, then dried over Na2SO4, filtered and concentrated to a yellow oil. The oil was purified by flash chromatography on Merck silica gel in 1% EtOAc in hexane. The desired fractions were combined and concentrated to afford a white solid, which was recrystallized from hexane to give product A as hard, off-white crystals (2.830 gm, 45%). The solution was cooled to room temperature an unexpectedly strong exothermic reaction The solution was then quenched with 10% Na2S2O3 in saturated NaHCO3 The solution was diluted with ether washed with 10% Na2S2O3 in saturated NaHCO3 (twice) and brine dried over Na2SO4 filtered concentrated to a yellow oil The oil was purified by flash chromatography on Merck silica gel in 1% EtOAc in hexane concentrated to afford a white solid, which was recrystallized from hexane

反応条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: An argon purged solution of the dibromide obtained in step (b) above (20.8 g, 75.4 mM) and anhydrous sodium acetate (NaOAc) (21.0 g) in ethyl acetate (370 ml) and glacial acetic acid (37 ml) was treated with 5% Pd/C (1.30 g) and the black suspension stirred under one atmosphere of H2 while monitoring uptake (22.4 l/mole). After 2.0 hours H2 uptake was complete, the mixture was filtered through Celite, the filtrate washed with saturated NaHCO3 and brine and then dried over anhydrous MgSO4 and evaporated to give the crude bromoester title product as a brown oil. The crude oil was combined with another batch (starting from 36.77 g of the dibromide) and vacuum distilled to give 25.771 g (61.3%) of the pure bromoester title product as a clear oil with b.p.=79°-80° C. (1.0 mm Hg) and with consistent C13NMR spectral data. TLC (1:1) ethyl acetatehexane, Rf=0.44, PMA. the mixture was filtered through Celite the filtrate washed with saturated NaHCO3 and brine dried over anhydrous MgSO4 evaporated

反応条件: temp:30.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: Methyl 4-bromo-3-oxobutyrate (300 mg), the above-mentioned wet bacterial cells (0.4 g), NADP+ (9 mg), glucose (750 mg), glucose dehydrogenase (manufactured by Amano Pharmaceuticals, Co., Ltd.) (1.2 mg), 100 mM phosphate buffer (pH 6.5) (15 ml) and butyl acetate (15 ml) are mixed. The mixture is stirred at 30° C. for 7 hr. During the stirring, 2M aqueous sodium carbonate solution is gradually added to adjust the pH of the reaction solution to 6.5±0.2. After the stirring has been finished, the reaction solution is centrifuged (1000×g, 5 min) to collect the organic layer. The organic layer is subjected to an analysis for content using gas chromatography under the following condition. The optical purity of methyl 4-bromo-3-hydroxybutyrate in the above-mentioned organic layer was also determined under the following condition. The organic layer was concentrated to give crude methyl (S)-4-bromo-3-hydroxybutyrate. (1000×g, 5 min) to collect the organic layer The organic layer was concentrated

反応条件: temp:92.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: A solution of (S)-4-Bromo-3-hydroxybutanoic acid, methyl ester (14.822 g, 75.2 mmol) in methyl ethyl ketone (255 mL) was treated with anhydrous sodium iodide (56.6 g, 376 mmol). The resulting red mixture was stirred for 23 hours under argon at a bath temperature of 92° C. The solution was allowed to cool to room temperature and the solids were filtered through celite. The receiver flask was switched and the celite bed was washed with ethyl acetate (3×50 mL). The original MEK product-rich filtrate was concentrated to approximately 60 mL. The residue was treated with the ethyl acetate wash from above followed by 250 mL of fresh ethyl acetate. The solution was washed with sodium sulfite solution (2×100 mL), brine (1×75 mL) and dried (sodium sulfate). Filtration followed by concentration in vacuo produced a yellow liquid which was evaporated several times from hexane and dried under high vacuum (5 mm) to 15.48 g (84%). to cool to room temperature the solids were filtered through celite the celite bed was washed with ethyl acetate (3×50 mL) The original MEK product-rich filtrate was concentrated to approximately 60 mL The residue was treated with the ethyl acetate wash The solution was washed with sodium sulfite solution (2×100 mL), brine (1×75 mL) dried (sodium sulfate) Filtration followed by concentration in vacuo produced a yellow liquid which was evaporated several times from hexane dried under high vacuum (5 mm) to 15.48 g (84%)