PROCESS FOR PRODUCING OPTICALLY ACTIVE (-)-2-HALO-1-(SUBSTITUTED PHENYL)ETHANOL

Abstract: A process for producing (-)-substituted styrene oxide efficiently, which comprises reducing 2-bromo-1-(3'-chlorophenyl)ethanone asymmetrically into (-)-2-bromo-1-(3'-chlorophenyl)ethanol in the presence of a microorganism belonging to any of the 11 genera including Ashbya and Brettanomyces , and cyclizing the isolated and purified product under an alkaline condition into (-)-substituted styrene oxide.

Description: The present invention relates to a method of manufacturing (-)-2-halo-1-(substituted phenyl) ethanol and more particularly to a method of efficiently manufacturing (-)-2-halo-1-(substituted phenyl) ethanol by bringing a microorganism into contact with 2-halo-1-(substituted phenyl) ethanone. These compounds are useful as materials for synthesis of drugs, agrichemicals et cetera requiring to be optically active. As to optically active (-)-2-halo-1-(substituted phenyl) ethanol, the present inventors have not yet seen any patent, report or the like relating to its manufacture. US-A 4,868,344 discloses a process for producing optically pure haloalcohols, e.g. optically pure (-)-2-chloro-1-(2,4-dichlorphenyl)-2-ethanol. EP-A 198 440 discloses a process for preparing optically active 2-halo-1-phenylethanol using microorganisms of the genera Candida, Geotrichum, Saccharomyces, Torulopsis and Rhodotorula, respectively. After having made intensive studies for development of an efficient method of manufacturing optically active (-)-2-halo-1-(substituted phenyl) ethanol the present inventors discovered existence of some microorganisms capable of asymmetrically reducing 2-halo-l-(substituted phenyl) ethanone stereospecifically and converting it into (-)-2-halo-1-(substituted phenyl) ethanol and thus completed the invention. The aspect of the present invention is to provide a method of bringing a 2-halo-1-(substituted phenyl) ethanone represented by a general formula [1] (where X represents a chlorine atom or a bromine atom and each of substitution groups R 1 , R 2 , R 3 represents a hydrogen atom, a chlorine atom, a fluorine atom, a methyl group or a methoxy group except cases where 3 substitution groups are all hydrogen atoms) into contact with a microorganism selected from those belonging to Ashbya genus, Brettanomyces genus, Candida genus, Cryptococcus genus, Pichia genus, Rhodosporidium genus and Trigonopsis genus capable of reducing it asymmetrically into (-)-2-halo-1-(substituted phenyl) ethanol represented by a general formula [2] (where X and substitution groups R 1 , R 2 , R 3 are the same as in the general formula [ 1 ] and * indicates an asymmetric carbon atom) and thereby to prepare (-)-2-halo-1-(substituted phenyl) ethanol. The microorganism used in the present invention for asymmetrical reduction of 2-halo-1-(substituted phenyl) ethanone for conversion into (-)-2-halo-1-(substituted phenyl) ethanol can be found in the following way. For example, 50 ml of A culture medium comprising 40 g of glucose, 3 g of yeast extract, 13 g of (NH 4 ) 2 HPO 4 , 7 g of KH 2 PO 4 , 0.8 g of MgSO 4 ·7H 2 O, 60 mg of ZnSO 4 ·7H 2 O, 90 mg of FeSO 4 ·7H 2 O, 5 mg of CuSO 4 5H 2 O, 10 mg of MnSO 4 ·4H 2 O and 0.1 g of NaCl (per liter) is put into a 500 ml Sakaguchi flask, a given microorganism is inoculated after sterilization and the culture medium is cultured under shaking for 2 days at 30 °C. Thereafter, cells are collected by centrifugal separation, suspended in 25 ml of 0.1M phosphate buffer solution (pH 7.0) containing 0.5% of 2-bromo-1-(3'-chlorophenyl) ethanone and 3% of glucose and shaken in a 500 ml Sakaguchi flask for 2-3 days at 30 °C. They are then admixed with an equivalent amount of ethyl acetate, followed by extraction, and the resulting (-)-2-bromo-1-(3'-chlorophenyl) ethanol is analyzed by gas chromatography (column: silicone OV-7,  0.3 x 200 cm, column temperature 190 °C, N 2 gas pressure 1.18 bar (1.2 kg/cm 2 )). The optical purity of (-)-2-bromo-1-(3'-chlorophenyl) ethanol can be determined by high-performance liquid chromatography (column: Nippon Bunko K.K.'s Chiralcel-OJ, dissolving solvent:hexane/isopropanol (50/1), flow rate:1.0 ml/min., detection:220 nm) after refining the extracted oil by distillation with the oil separated in 44.8 minutes of retention time for (-) isomer and 54.9 minutes of retention time for (+) isomer. As microorganism may be used any of the known genuses capable of asymmetrical reduction of 2-halo-1-(substituted phenyl) ethanone for conversion into (-)-2-halo-1-(substituted phenyl) ethanol. As such may be exemplified, among others, Ashbya gossypii IFO 0560, Brettanomyces custersianus IFO 1585, Candida humicola CBS 2774, Candida intermedia IFO 0761, Candida krusei IFO 0011, Candida magnoliae IFO 0705, Candida pinus IFO 0741, Candida saitoana IFO 0768, Candida sake CBS 2219, Candida tropicalis IFO 1403, Cryptococcus albidus IFO 0378, Cryptococcus terreus IFO 0727, Geotrichum hirtum CBS 189,53, Geotrichum loubieri CBS 252,61, Pichia farinosa IFO 0574, Pichia membranaefaciens IFO 0460, Rhodosporidium toruloides IFO 0871, Rhodotorula glutinis IFO 1099, Rhodotorula glutinis var. dairenensis IFO 0415, Rhodotorula graminis IFO 0190, Rhodotorula minuta IFO 0387, Rhodotorula rubra IFO 0383, Saccharomyces cerevisiae IFO 0614 and Trigonopsis variabilis IFO 0671. For the cultivation of these microorganisms may be used usually any nutriment which they can assimilate. For the cultivation thereof, therefore, may be used an ordinary culture media containing proper amounts of, for example, carbohydrates such as glucose and sucrose, alcohols such as ethanol and glycerol, hydrocarbons such as paraffin, organic acids such as acetic acid and propionic acid, carbon sources such as soy bean oil and mixtures thereof, yeast extracts, peptone, meat extracts, corn steep liquor, nitrogen-containing inorganic and organic nutriments such as ammonium sulfate and ammonia; inorganic nutriments such as phosphates, magnesium, iron, manganese and potassium, and vitamins such as biotin and thiamine. Cultivation is carried out aerobically for 1-5 days at 20-40°C with pH of the nutrient medium adjusted to 4.0-9.5. As reducing method are known, for instance, a method in which a culture solution is used as it is and a method in which cells are separated by centrifugal separation and resuspended in, for example, a phosphate buffer solution or water and then admixed with 2-halo-1-(substituted phenyl) ethanone for the reaction to proceed. For the reaction a carbon source such as glucose or sucrose may be added to the culture medium as an energy source. The cells may be used as live cells but may as well be used after acetone treatment or freeze drying. Such cells may also be used fixed on a carrier. 2-halo-1-(substituted phenyl) ethanone may be added as it is or after dissolution in an organic solvent lest it should interfere with the progress of the reaction either en bloc at the start of the reaction or in portions as the reaction proceeds. The reaction may be conducted under stirring for 3-120 hours at a temperature of 10-60 °C with proper pH adjustment in a range of 5-9. Separation of (-)-2-halo-1-(substituted phenyl) ethanol as the reaction product may be done directly from the reaction liquor or after separation of cells followed by extraction by the use of a solvent such as ethyl acetate and dichlormethane, dehydration and subsequent refining by distillation or silica gel chromatography, and in this way (-)-2-halo-1-(substituted phenyl) ethanol of a high purity can be obtained with ease. Its optical purity can be determined in the same way as described above by high-performance liquid chromatography by the use of column, Chiral cel-OJ and hexane/isopropanol (30-50/1) as eluant. The (-)-2-halo-1-(substituted phenyl) ethanol thus obtained can be easily cyclized by heating or leaving at the room temperature under coexistence of an alkali such as NaOH in a quantity of not less than the equivalent mol and can be converted into (-)-substituted styrene oxide. Hereinafter the present invention will be described in greater detail with reference to preferred examples but it is to be understood that the present invention is by no means limited thereby. Unless otherwise noted, [%] in the following description means [weight %]. Example 1 50 ml of the aforementioned A culture medium was put into a 500 ml Sakaguchi flask and after sterilization the microorganisms shown in Table 1 were inoculated thereon, and aerobical cultivation was carried out under shaking for 2 days at 30°C. Cells were taken from the culture solution by centrifugal separation, 0.5% of 2-bromo-1-(3'-chlorophenyl) ethanone was suspended in 25 ml of 0.1M phosphate buffer solution (pH 7.0) containing 0.3% glucose and the reaction was carried out under shaking in the 500 ml Sakaguchi flask for 48 hours at 30 °C. After completion of the reaction, (-)-2-bromo-1-(3'-chlorophenyl) ethanol was extracted twice using the equivalent amount of ethyl acetate and the resulting ethyl acetate layer was analyzed by gas chromatography for determination of the degree of conversion. Then, after dehydration of ethyl acetate with Glauber's salt anhydride, the solvent was removed and (-)-2-bromo-1-(3'-chlorophenyl) ethanol was obtained. This was dissolved in methylene chloride and its optical purity was determined by high-performance liquid chromatography. The results were as shown in Table 1. Microorganisms (-)-2-bromo-1-(3'-chlorophenyl) ethanol Yield(%) Optical purity(%) Ashbya gossypii IFO 0560 55 72 Brettanomyces custersianus IFO 1585 10 100 Candida humicola CBS 2774 40 90 Candida intermedia IFO 0761 54 100 Candida krusei IFO 0011 31 100 Candida magnoliae IFO 0705 80 81 Candida pinus IFO 0741 44 92 Cryptococcus albidus IFO 0378 59 92 Cryptococcus terreus IFO 0727 63 98 Pichia farinosa IFO 0574 44 81 Pichia membranaefaciens IFO 0460 21 100 Rhodosporidium toruloides IFO 0871 79 100 Rhodotorula glutinis IFO 1099 56 67 Rhodotorula glutinis var. dairenensis IFO 0415 75 100 Rhodotorula graminis IFO 0190 51 100 Rhodotorula minuta IFO 0387 8 99 Rhodotorula rubra IFO 0383 56 100 Saccharomyces cerevisiae IFO 0614 21 76 Trigonopsis variabilis IFO 0671 76 88 Example 2 Rhodotorula glutinis var. dairenensis IFO 0415 was inoculated on 3 liters of A culture medium in a 5-liter minijar fermentator and cultivated for 24 hours under stirring (500 rpm.) at 30°C with air being introduced at a rate of 1 vvm. After completion of cultivation, cells were collected by centrifugal separation and 7.5 g of 2-bromo-1-(3'-chlorophenyl) ethanone and 38 g of glucose were added and the reaction was carried out for 24 hours at 30 °C under stirring (150 rpm.), the pH being adjusted to 7.0 by the use of NaOH. After the completion of the reaction, the reaction product was extracted twice using 750 ml of ethyl acetate. The ethyl acetate layer was first dehydrated using Glauber's salt anhydride, then the solvent was removed under reduced pressure and thus 5.2 g of an oily substance was obtained. This was distilled (130 °C 3.95 mbar (3 mmHg)) and there was obtained 3.9 g of colorless, oily (-)-2-bromo-1-(3'-chlorophenyl) ethanol. Its specific rotation [ α ] 20 / D was -25.5° (c = 1.02,CH 3 OH) and its optical purity was 100% e.e. as it was determined by high-performance liquid chromatography. H-NMR (90 MHz, CDCl 3 ) δ ppm 2.88 (br.s, 1H), 3.35-3.90 (m, 4H), 4.90 (d.d, J=315, 8Hz, 1H) 6.98-7.51 (m, 4H) Example 3 Bio-reaction and analysis were carried out in the same way as described in Example 1 except that the microorganisms used were those shown in Table 2 and as substrate was used 2-bromo-1-(2'-chlorophenyl) ethanone instead of 2-bromo-1-(3'-chlorophenyl) ethanone and as the reaction product (-)-2-bromo-1-(2'-chlorophenyl) ethanol was obtained. The results were as shown in Table 2. Microorganisms (-)-2-bromo-1-(2'-chlorophenyl) ethanol Yield(%) Optical purity(%) Candida humicola CBS 2774 23 92 Candida intermedia IFO 0761 23 92 Candida krusei IFO 0011 5 100 Candida magnoliae IFO 0705 98 96 Candida pinus IFO 0741 32 96 Candida saitoana IFO 0768 13 91 Candida sake IFO 2219 31 96 Candida tropicalis IFO 1403 32 86 Cryptococcus albidus IFO 0378 15 98 Cryptococcus terreus IFO 0727 39 100 Geotrichum hirtum CBS 189,53 75 92 Geotrichum loubioeri CBS 252,61 27 92 Pichia farinosa IFO 0574 34 88 Rhodosporidium toruloides IFO 0871 62 98 Rhodotorula glutinis IFO 1099 87 100 Rhodotorula glutinis var. dairenensis IFO 0415 15 100 Rhodotorula graminis IFO 0190 31 98 Rhodotorula rubra IFO 0383 17 100 Trigonopsis variabilis IFO 0671 7 99 Example 4 Bio-reaction and analysis were conducted in the same way as described in Example 2 except that as microorganism was used Rhodotorula glutinis IFO 1099 and as substrate was used 2-bromo-1-(2'-chlorophenyl) ethanone and as the reaction product was obtained 4.2 g of (-)-2-bromo-1-(2'-chlorophenyl) ethanol. Its specific rotation [ α ] 20 / D was 41.5 ° (C=1.02, CH 3 OH) and its optical purity as determined by high-performance liquid chromatography was 100% e.e. H-NMR (90MHz, CDCl 3 ) δ ppm 2.78-3.06 (m, 1H), 3.39 (d.d, J=9, 10Hz, 1H), 3.73 (d.d, J=2.5, 10Hz, 1H), 5.04-5.39 (m, 1H), 6.68-7.68 (m, 4H) Example 5 Bio-reaction and analysis were carried out in the same way as described in Example 2 except that as microorganisms were used those shown in Table 3 and as substrate was used 2-bromo-1-(4'-chlorophenyl) ethanone instead of 2-bromo-1-(3'-chlorophenyl) ethanone and as the reaction product (-)-2-bromo-1-(4'-chlorophenyl) ethanol was obtained. The results were as shown in Table 3. Microorganisms (-)-2-bromo-1-(4'-chlorophenyl) ethanol Yield(%) Optical purity(%) Ashbya gossypii IFO 0560 47 95 Burettanopmyces custersianus IFO 1585 11 100 Candida humicola CBS 2774 51 73 Candida intermedia IFO 0761 26 72 Candida krusei IFO 0011 8 87 Candida magnoliae IFO 0705 89 68 Candida pinus IFO 0741 25 96 Candida saitoana IFO 0768 55 98 Candida tropicalis IFO 1403 21 90 Cryptococcus alibidus IFO 0378 24 97 Cryptococcus terreus IFO 0727 9 100 Pichia falinosa IFO 0574 51 84 Rhodosporidium toruloides IFO 0871 17 88 Rhodotorula glutinis IFO 1099 55 93 Rhodotorula glutinis var. direnensis IFO 0415 67 100 Rhodotorula graminis IFO 0190 31 89 Rhodotorula rubra IFO 0383 18 94 Trigonopsis variabilis IFO 0671 42 76 Example 6 3io-reaction and analysis were carried out in the same way as described in Example 2 except that as substrate was used 2-bromo-1-(4'-chlorophenyl) ethanone instead of 2-bromo-1-(3'-chlorophenyl) ethanone and as the reaction product was obtained 3.6 g of (-)-2-bromo-1-(4'-chlorophenyl) ethanol. Its boiling point was 115-120 °C / 5.26 mbar (4 mmHg), specific rotation [ α ] 20 / D was 26.6 ° (C=1.10, CH 3 OH) and its optical purity as determined by high-performance chromatography was 100% e.e. H-NMR (90MHz, CDCl 3 ) δ ppm 2.77 (br,s, 1H), 3.18-3.70 (m, 2H), 4.82 (d.d, J=3.5, 7.5Hz, 1H), 6.82-7.44 (m, 4H) Example 7 Bio-reaction and analysis were carried out in the same way as described in Example 2 except that as substrate was used 3.25 g each of 2-chloro-1-(2'-chlorophenyl) ethanone, 2-chloro-1-(3'-chlorophenyl) ethanone and 2-chloro-1-(4'-chlorophenyl) ethanone instead of 2-bromo-1-(3'-chlorophenyl) ethanone and as the reaction products (-)-2-chloro-1-(2'-chlorophenyl) ethanol, (-)-2-chloro-1-(3'-chlorophenyl) ethanol and (-)-2-chloro-1-(4'-chlorophenyl) ethanol were obtained respectively. The yields and physical properties of the respective reaction products were as,shown in Table 4. Substrate 2-chloro-1-(2'-chlorophenyl)ethanone 2-chloro-1-(3'-chlorophenyl)ethanone 2-chloro-1-(4'-chlorophenyl)ethanone Reaction product (-)-2-chloro-1-(2'-chlorophenyl) ethanol (-)-2-chloro-1-(3'-chlorophenyl) ethanol (-)-2-chloro-1-(4'-chlorophenyl) ethanol Yield 1.7 g 1.4 g 1.3 g Boil.point 150 °C / 6,58 mbar (5 mmHg) 130 °C / 3,95-5,26 mbar (3-4 mmHg) 140°C / 6,58 mbar (5 mmHg) Specific rotation [α] ] 20 / D (C=1, CH 3 OH) -55.6 ° -33.8 ° -36.1 ° Optical purity 100% 100% 100% H-NMR (90MHz CDCl 3 ) δ ppm 3.29(br,S,1H) 3.50(dd,J=11.5, 8.5Hz,1H) 3.85(dd,J=11.5, 3Hz,lH) 5.30(dd,J=11.5, 3Hz,1H) 7.07-7.75(m,4H) 2.69(br,S,1H) 3.27-3.90(m,2H) 4.88(dd,J=3.5, 8Hz,1H) 7.15-7,54(m,4H) 2.27(br,S,1H) 3.41-3.82(m,2H) 4.83(dd,J=4.5, 7.5Hz,1H) 7.11-7.50(m,4H) The compounds of the following examples 8 to 16 do not fall under the scope of the claims. Example 8 10 g each of (-)-2-halo-1-(chloro-substituted phenyl) ethanols obtained in Examples 2, 4, 6 and 7 was admixed with 5 ml of 40% NaOH aqueous solution and 10 ml of methylene chloride and was caused to react for 6 hours at 50°C. After cooling, the reaction product was admixed with 20 ml of methylene chloride, the methylene chloride layer was washed with saturated brine and after dehydration and filtration, methylene chloride was removed under reduced pressure and an oily epoxide was thus obtained. This was refined by distillation under reduced pressure and each (-)-chloro-substituted styrene oxide was obtained as shown in Table 5. (-)-2-bromo-1-(3'-chlorophenyl)ethanol (-)-2-chloro-1-(3'-chlorophenyl)ethanol (-)-2-bromo-1-(2'-chlorophenyl)ethanol (-)-2-bromo-1-(4'-chlorophenyl)ethanol (-)-chloro-substituted styrene (-)-3'-chlorostyrene oxide (-)-2'-chlorostyrene oxide (-)-4'-chlorostyrene oxide oxide Yield 6.2 g 7.1 g 5.4 g 5.7 g Boil.point 90-93°C / 6,58 mbar (5 mmHg) 90-93°C / 6,58 mbar (5 mmHg) 80-85°C / 5,26 mbar (4 mmHg) 85-90 °C / 5,26 mbar (4 mmHg) [ α ] 20 / D (C=1,CHCl 3 ) -11.8 ° -11.8 ° -67.5 ° -25.5 ° H-NMR (90MHz CDCl 3 ) δ ppm 2.75(dd,J=2.5,6Hz,1H) 3.13(dd,J=3.5,6Hz,1H) 3.84(dd,J=2.5,3.5Hz,lH) 7.10-7.45(m,4H) 2.63(dd,J=2.5 6Hz,1H) 3.15(dd,J=3, 6Hz,1H) 4.17(dd,J=2.5 4Hz,1H) 7.01-7.51(m,4H) 2.75(dd,1H) 3.08(dd,1H) 3.77(dd,1H) 6.99-7.42 (m,4H) Example 9 500 ml of the aforementioned A culture medium was put into a 2-liter Sakaguchi flask, Rhodotorula glutinis IFO 0415 was inoculated thereon, cultivation was done in the same way as described in Example 1, the resulting cells were collected, 1.5 g of 2-chloro-(4'-fluorophenyl) ethanone was suspended in 300 ml of 0.1M phosphate buffer solution (pH 7.0) containing 0.3% glucose, the suspension was put into the 2-liter Sakaguchi flask and it was reacted under shaking for 48 hours at 30°C. After the reaction was over, the reaction product was extracted twice using 300 ml of ethyl acetate. After dehydration of the ethyl acetate layer with Glauber's salt anhydride, the solvent was removed under reduced pressure and thus an oily substance was obtained. It was then distilled (105 °C / 5,26 mbar (4 mmHg)) and 1.3 g of (-)-2-chloro-1-(4'-fluorophenyl) ethanol was obtained as a colorless oily substance. its specific rotation [α] 20 / D was 38.81° (c=1,01,CH 3 OH) and its optical purity was 100% e.e. as it was determined by means of high-performance liquid chromatography. 1 g of this (-)-2-chloro-1-(4'-fluorophenyl) ethanol was then admixed with the equivalent mol of 40% aqueous solution of NaOH and 5 ml of methylene chloride and the mixture was caused to react for 6 hours at 50 °C. After cooling, 5 ml of methylene chloride was added, the methylene chloride layer was washed with saturated brine and after dehydration and filtration, methylene chloride was removed under reduced pressure and crude epoxide was obtained as an oily substance. This was refined under reduced pressure (85°C, 5.26 mbar (4 mmHg)) and 0.65 g of (-)-4'-fluorostyrene oxide was obtained in a colorless oily form. Its specific rotation [α] 20 / D was -20.96 ° (c=1.04,CHCl 3 ). (-)-2-chloro-1-(4'-chlorophenyl) ethanol: H-NMR(90MHz, CDCl 3 ) δ ppm 3.00 (s, 1H), 3.40-3.83 (m, 2H), 4.85 (d.d, J=4.5, 7.5Hz, 1H), 6.83-7.53 (m, 4H) (-)-4'-fluorostyrene oxide: H-NMR (90 MHz, CDCl 3 ) δ ppm 2.72 (d.d, J=2.5, 6.0Hz, 1H), 3.08 (d.d, J=4.5, 6.0Hz, 1H), 3.82 (d.d. J=2.5, 4.0Hz, 1H), 6.85-7.45 (m, 4H) Example 10 Cultivation, reaction and purification were carried out in the same way as described in Example 9 except that 2-chloro-1-(2', 4'-dichlorophenyl) ethanone, 2-chloro-1-(3', 4'-dichlorophenyl) ethanone, 2-chloro-1-(2', 5'-dichlorophenyl) ethanone were used as substrate, and (-)-2-chloro-1-(2', 4'-dichlorophenyl) ethanol, (-)-2-chloro-1-(3', 4'-dichlorophenyl) ethanol, (-)-2-chloro-1-(2', 5'-dichlorophenyl) ethanol was obtained. The yield, specific rotation and optical purity determined by high-performance liquid chromatography were as shown in Table 6. Then, epoxidated in the same way as Example 9, (-)-2', 4'-dichlorostyrene oxide, (-)-3', 4'-dichlorostyrene oxide, (-)-2', 5'-dichlorostyrene oxide were obtained. The yield and the specific rotation were as shown in Table 6. Substrate 2-chloro-1-(2',4'-dichlorophenyl)ethanone 2-chloro-1-(3',4'-dichlorophenyl)ethanone 2-chloro-1-(2',5'-dichlorophenyl) ethanone Reaction product (-)-2-chloro-1-(2',4'-dichloro-phenyl)ethanol (-)-2-chloro-1-(3',4'-dichloro-phenyl)ethanol (-)-2-chloro-1-(2',5'-dichloro-phenyl)ethanol Yield 0.7 g 0.8 g 0.85 g Form Colorless oil Colorless oil Colorless oil Boil.point Specific rotation 160 °C / 5.26 mbar (4 mmHg) -51.37 ° 170 °C / 5.26 mbar (4 mmHg) -32.72 ° 140 °C / 5.26 mbar (4 mmHg) -43.40 ° [α] 20 / D (C=1,CH 3 OH) Optical 100% e.e. 100% e.e. 100% e.e. purity H-NMR (90MHz CDCl 3 ) δ ppm 2.86(br,S,1H) 3.46-3.88(m,2H) 4.96(dd,J=4.5, 7.0Hz,1H) 7.13-7.70(m,4H) 2.95(br,S,1H) 3.48(dd,J=8.5 11.5Hz,1H) 3.85(dd,J=3.0, 11.4Hz,1H) 5.18(dd,J=3.0, 8.5Hz,1H) 7.18-7.65(m,3H) 2.98(br,S,1H) 3.48(dd,J=9.0. 12.0Hz,1H) 3.85(dd,J=3.0, 11.OHz,1H) 5.25(dd,J=3,0, 8.5Hz,1H) 7.35-7.59(m,2H) Reaction product (-)-2',4'-dichloro styrene oxide (-)-3',4'-dichloro styrene oxide (-)-2',5'-dichloro styrene oxide Yield 0.61 g 0.65 g 0.68 g Form Colorless oil Colorless oil Colorless oil Boil.point Specific rotation 120 °C / 5.26 mbar (4 mmHg) 125 °C / 5.26 mbar (4 mmHg) 110°C / 5.26 mbar (4 mmHg) [ α ] 20 / D -61.30 ° -18.30 ° -22.15 ° (C=1,CH 3 OH) H-NMR (90MHz CDCl 3 ) δ ppm 2.61(dd,J=2.5, 5.5 Hz,1H) 3.17(dd,J=3.0, 5.5Hz,1H) 4.12(dd,J=2.5, 3.5Hz,1H) 7.05-7.41(m,3H) 2.73(dd,J=2.5, 5.5Hz,1H) 3.12(dd,J=4.0, 5.5Hz,1H) 3.80(dd,J=2.5, 4.0Hz,1H) 7.02-7.05(m,3H) 2.66(dd,J=3.0, 6.0Hz,1H) 3.21(dd,J=5.5, 4.5Hz,1H) 4.17(dd,J=3.0, 4.5Hz,1H) 7.16-7.46(m,3H) Example 11 Cultivation and reaction were carried out in the same way as described in Example 9 except that 2-chloro-1-(2',3',4'-trichlorophenyl) ethanone was used as substrate and, after the reaction was over, extraction was done twice using 300 ml ethyl acetate. The ethyl acetate layer was dehydrated with Glauber's salt anhydride, this followed by removal of solvent under reduced pressure, and an oily substance was obtained. It was then refined by silica gel chromatography (hexane/ethyl acetate=7/1) and 1.1 g of a colorless oil of (-)-2-chloro-1-(2',3',4'-trichlorophenyl) ethanol was obtained. Its specific rotation [ α ] 20 / D was -52.29 ° (C=1.07, CH 3 OH) and its optical purity was 100% e.e. as it was determined by high-performance liquid chromatography. (-)-2'-chloro-1-(2',3',4'-trichlorophenyl) ethanol H-NMR(90MHz, CDCl 3 ) δ ppm 3.10(br,s,1H), 3.50(d.d, J=9.0, 11.0Hz,1H) 3.85(d.d,J=3.0, 11.0Hz,1H) 5.25(d.d,J=3.0, 8.5Hz,lH) 7.35-7.59(m,2H) Example 12 Cultivation, reaction and purification were carried out in the same way as described in Example 9 except that 2-chloro-1-(2'-methylphenyl) ethanone, 2-chloro-1-(3'-methylphenyl) ethanone, 2-chloro-1-(4'-methylphenyl) ethanone were used as substrate, and (-)-2-chloro-1-(2'-methylphenyl) ethanol, (-)-2-chloro-1-(3'-methylphenyl) ethanol, (-)-2-chloro-1-(4'-methylphenyl) ethanol were obtained. The yield, specific rotation and optical purity determined by high-performance liquid chromatography were as shown in Table 7. Then, the reaction product was epoxidated in the same way as Example 9, and (-)-2'-methylstyrene oxide, (-)-3'-methylstyrene oxide, (-)-4'-methylstyrene oxide were obtained. The yield and the specific rotation were as shown in Table 7. Substrate 2-chloro-1-(2'-methylphenyl)ethanone 2-chloro-1-(3'-methylphenyl)ethanone 2-chloro-1-(4'-methylphenyl)ethanone Reaction product (-)-2-chloro-1-(2'-methylphenyl)ethanol (-)-2-chloro-1-(3'-methylphenyl)ethanol (-)-2-chloro-1-(4'-methylphenyl)ethanol Yield 1.3 g 1.2 g 1.3 g Form Colorless oil Colorless oil Colorless oil Boil.point Specific 110 °C / 5.26 mbar (4 mmHg) 115 °C / 5.26 mbar (4 mmHg) 120 °C / 5.26 mbar (4 mmHg) rotation [ α ] 20 / D (C=1,CH 3 OH) -56.96 ° -40.39 ° -42.58 ° Optical purity 100% e.e. 100% e.e. 100% e.e. H-NMR (90MHz CDCl 3 ) δ ppm 2.30(br,3H) 2.72(br,S,1H) 3.37-3.77(m,2H) 5.03(dd,J=4.5, 8.5Hz,1H) 7.01-7.61(m,3H) 2.35(S,3H) 2.60(br,S,1H) 3.46-3.85(m,2H) 4.82(dd,J=5.0 7.5Hz,1H) 7.03-7.42(m,4H) 2.35(S,3H) 2.68(br,S,1H) 3.46-3.85(m,2H) 4.85(dd,J=5.0 7.5Hz,1H) 7.00-7.40(m,4H) Reaction product Yield (-)-2'-methylstyrene oxide 0.8 g (-)-3'-methylstyrene oxide 0.8 g (-)-4'-methylstyrene oxide 0.8 g Form Colorless oil Colorless oil Colorless oil Boil.point Specific 70 °C / 5.26 mbar (4 mmHg) 75 °C / 5.26 mbar (4 mmHg) 80°C / 5.26 mbar (4 mmHg) rotation [ α ] 20 / D (C=1,CH 3 OH) -80.59 ° -23.01 ° -29.46 ° H-NMR CDCl 3 ) δ ppm 2.39(br,3H) 2.63(dd,J=2.5, 6.OHz,1H) 3.09(dd,J=3.0, 5.5Hz,1H) 3.93(dd,J=2.5, 3.0Hz,1H) 7.15(S,4H) 2.30(S,3H) 2.69(dd,J=2.5, 6.0Hz,1H) 3.03(dd,J=3.0, 6.0Hz,1H) 3.74(dd,J=2.5, 3.0Hz,1H) 6.89-7.29(m,4H) 2.30(S,3H) 2.73(dd,J=2.5, 5.5Hz,1H) 3.03(dd,J=4.5, 5.5Hz,1H) 3.74(dd,J=2.5, 4.5Hz,1H) 7.12(S,4H) Example 13 Cultivation, reaction and purification were carried out in the same way as described in Example 9 except that 2-chloro-1-(2',4'-dimethylphenyl) ethanone and 2-chloro-1-(3', 4'-dimethylphenyl) ethanone were used as substrate, and (-)-2-chloro-1-(2', 4'-dimethylphenyl) ethanol and (-)-2-chloro-1-(3', 4'-dimethylphenyl) ethanol were obtained. The yield, specific rotation and optical purity determined by high-performance liquid chromatography were shown in Table 8. Then, epoxidated in the same way as Example 9, (-) -2', 4'-dimethylstyrene oxide and (-)-3', 4'-dimethylstyrene oxide were obtained. The yield and the specific rotation were as shown in Table 8. Substrate 2-chloro-1-(2',4'-dimethylphenyl)ethanone 2-chloro-1-(3',4'-dimethylphenyl)ethanone Reaction product (-)-2-chloro-1-(2',4'-dimethylphenyl)ethanol (-)-2-chloro-1-(3',4'-dimethylphenyl)ethanol Yield 1.3 g 1.3 g Form Colorless oil Crystal (mp. 61.5°C ) Boil.point Specific 130 °C / 5.26 mbar (4 mmHg) 125°C / 5.26 mbar (4 mmHg) rotation -39.80 ° -49.37 ° [ α ] 20 / D (C=1,CH 3 OH) Optical purity 100% e.e. 100% e.e. H-NMR (90MHz CDCl 3 ) δ ppm 2.31(S,6H) 2.59(br,S,1H) 3.45-3.85(m,2H) 5.08(dd,J=4.0,9.0Hz,1H) 6.90-7.56(m,3H) 2.28(S,6H) 2.55(br,S,1H) 3.60-3.80(m,2H) 4.85(dd,J=4.5,7.5Hz,1H) 7.00-7.31(m,3H) Reaction product (-)-2',4'-dimethylstyrene oxide (-)-3',4'-dimethylstyrene oxide Yield 0.76 g 0.73 g Form Colorless oil Colorless oil Boil.point Specific 90 °C / 5.26 mbar (4 mmHg) 88°C / 5.26 mbar (4 mmHg) rotation [ α ] 20 / D (C=1,CH 3 OH) -69.14 ° -29.03 ° H-NMR (90MHz CDCl 3 ) δ ppm 2.28(S,3H) 2.35(S,3H) 2.65(dd,J=3.0,6.0Hz,1H) 3.08(dd,J=4.0,6.0Hz,1H) 3.98(dd,J=3.0,4.0Hz,1H) 6.83-7.26(m,3H) 2,22(S,6H) 2.73(dd,J=2.5,5.5Hz,1H) 3.05(dd,J=4.0,5.5Hz,1H) 3.75(dd,J=2.5,4.0Hz,1H) 6.88-7.27(m,3H) Example 14 Cultivation, reaction and purification were carried out in the same way as described in Example 9 except that 2-chloro-1-(2'-methoxyphenyl) ethanone, 2-chloro-1-(3'-methoxyphenyl) ethanone, 2-chloro-1-(4'-methoxyphenyl) ethanone were used as substrate and (-)-2-chloro-1-(2'-methoxyphenyl) ethanol, (-)-2-chloro-1-(3'-methoxyphenyl) ethanol and (-)-2'-chloro-1-(4'-methoxyphenyl) ethanol were obtained. The yield, specific rotation and optical purity determined by high-performance liquid chromatography were shown in Table 9. Then, by epoxidation in the same way as Example 9 (-)-2'-methoxystyrene oxide, (-)-3'-methoxystyrene oxide and (-)-4'-methoxystyrene oxide were obtained. The yield and the specific rotation were as shown in Table 9. Substrate 2-chloro-1-(2'-methoxyphenyl)ethanone 2-chloro-1-(3'-methoxyphenyl)ethanone 2-chloro-1-(4'-methoxyphenyl)ethanone Reaction product (-)-2-chloro-1-(2'-methoxyphenyl)ethanol (-)-2-chloro-1-(3'-methoxyphenyl)ethanol (-)-2-chloro-1-(4'-methoxyphenyl)ethanol Yield 1.4 g 1.4 g 1.4 g Form Colorless oil Colorless oil Colorless oil Boil.point Specific 135 °C / 5.26 mbar (4 mmHg) 140°C / 5.26 mbar (4 mmHg) 145°C / 5.26 mbar (4 mmHg) rotation [ α ] 20 / D (C=1,CH 3 OH -68.25 ° -34.74 ° -41.50 ° Op.purity H-NMR (90MHz CDCl 3 ) δ ppm 100% e.e. 3.00(br,S,1H) 3.40-4.00(m,2H) 3.83(S,3H) 5.12(dd,J=5.5, 8.5Hz,1H) 6.74-7.56(m,4H) 100% e.e. 2.80(br,S,1H) 3.78(S,3H) 3.43-3.90(m,2H) 4.83(dd,J=4.5, 7.5Hz,1H) 6.73-7.47(m,4H) 2.82(br,S,1H) 3.36-3.90(m,2H) 3.78(S,3H) 4.85(dd,J=5.5, 7.5Hz,1H) 6.93(d,J=9.0, Hz,2H) 7.33(d,J=9.0, Hz,2H) Reaction product (-)-2'-methoxystyrene oxide (-)-3'-methoxystyrene oxide (-)-4'-methoxystyrene oxide Yield 0.7 g 0.7 g 0.7 g Form Colorless oil Colorless oil Colorless oil Boil.point Specific 110 °C / 5.26 mbar (4 mmHg) 115°C / 5.26 mbar (4 mmHg) 120°C / 5.26 mbar (4mmHg) rotation [ α ] 20 / D (C=1,CH 3 OH) -58.95 ° -14.95 ° -23.00 ° H-NMR (90MHz CDCl 3 ) δ ppm - 2.68(dd,J=2.5, 6.0Hz,1H) 3.12(dd,J=3.0, 6.0Hz,1H) 3.85(S,3H) 4.20(dd,J=2.5, 3.0Hz,1H) 6.76-7.43(m,4H) 2.73(dd,J=2.5, 6.0Hz,1H) 3.05(dd,J=5.5, 6.0Hz,1H) 3.75(S,3H) 3.56-3.92(m,1H) 6,61-7.40(m,4H) 2.75(dd,J=2.5, 5.5Hz,1H) 3.04(dd,J=4.5, 5.5Hz,1H) 3.75(S,3H) 3.47-3.87(m,1H) 6.83(d,J=9.0, Hz,2H) 7.16(d,J=9.0, Hz,2H) Example 15 Cultivation, reaction and purification were carried out in the same way as described in Example 9 except that 2-chloro-1-(2',5'-dimethoxyphenyl) ethanone, 2-chloro-1-(3',4'-dimethoxyphenyl) ethanone were used as substrate and (-)-2-chloro-1-(2',5'-dimethoxyphenyl) ethanol, (-)-2-chloro-1-(3',4'-dimethoxyphenyl) ethanol were obtained. The yield, specific rotation and optical purity determined by high-performance liquid chromatography were shown in Table 10. Then, by epoxidation of (-)-2-chloro-1-(2',5'-dimethoxyphenyl) ethanol in the same way as Example 9 (-)-2',5'-dimethoxystyrene oxide was obtained. The yield and the specific rotation were as shown in Table 10. Substrate 2-chloro-1-(2',5'-dimethoxyphenyl)ethanone 2-chloro-1-(3',4'-dimethoxyphenyl)ethanone Reaction product (-)-2-chloro-1-(2',5'-dimethoxyphenyl)ethanol (-)-2-chloro-1-(3',4'-dimethoxyphenyl)ethanol Yield 1,4 g 1.3 g Form Colorless oil Colorless oil Boil.point Specific rotation 150°C / 5.26 mbar (4 mmHg) -51.27 ° 175°C / 5.26 mbar (4 mmHg) -25.59 ° [α] 20 / D (C=1,CH 3 OH) Optical purity 100% e.e. 100% e.e. H-NMR (90MHz CDCl 3 ) δ ppm 3.16(br,S,1H) 3.73(S,3H) 3.77(S,3H) 3.40-3.93(m,2H) 5.08(dd,J=4.0,8.0Hz,lH) 6.60-7.10(m,3H) 2.80(br.S,1H) 3.85(S,6H) 3.40-3.80(m,2H) 4.75(dd,J=6.0,7.5Hz,1H) 6.60-7.05(m,3H) Reaction product (-)-2'-5'-dimethoxystyrene oxide Yield 0.7 g Form Colorless oil Boil.point 140°C / 5.26 mbar (4 mmHg) Specific rotation -33.47 ° [ α ] 20 / D (C=1,CH 3 OH) H-NMR (90MHz CDCl 3 ) δ ppm 2.64(dd,J=2.5,6.0Rz,1H) 3.08(dd,J=4.0,6,0Hz,1H) 3.72(S,3H) 3.79(S,3H) 4.15(dd,J=2.5,4.0Hz,1H) 6.55-7.05(m,3H) Example 16 Cultivation and reaction were carried out in the same way as described in Example 9 except that 2-chloro-1-(2',3',4'-trimethoxyphenyl) ethanone, 2-chloro-1-(3',4',5'-trimethoxyphenyl) ethanone were used as substrate and after completion of reaction, extraction was done twice using 300 ml of ethyl acetate. After dehydration of the ethyl acetate layer, the solvent was removed under reduced pressure, the resulting oily substance was refined by silica gel chromatography (hexane/ethyl acetate = 7/1) and (-)-2-chloro-1-(2',3',4'-trimethoxyphenyl) ethanol, (-)-2-chloro-1-(3',4',5'-trimethoxyphenyl) ethanol were obtained. Their yield and specific rotation were as shown in Table 11. Substrate 2-chloro-1-(2',3',4'-trimethoxyphenyl) ethanone 2-chloro-1-(3',4',5'-trimethoxyphenyl) ethanone Reaction product (-)-2-chloro-1-(2', 3',4'-trimethoxyphenyl) ethanol (-)-2-chloro-1-(3', 4'.5'-trimethoxyphenyl) ethanol Yield 1.2 g 1.3 g Form Colorless oil Crystal (mp. 65.5 °C ) Specific rotation [ α ] 20 / D (C=1,CH 3 OH) -22.05 ° -24.76 ° Optical purity - - H-NMR (90MHz CDCl 3 ) δ ppm 2.89(br,S,1H) 3.86(S,6H) 3.96(S,3H) 3.42-4.23(m,2H) 5.08(dd,J=3.5,7.5Hz,1H) 6.71(d,j=9.0,Hz,1H) 7.16(d,J=9.0,Hz,1H) 2.82(br,S,1H) 3.36-3.76(m,2H) 3.80(S,3H) 3.84(S,6H) 4.80(dd,J=4.5,7,5Hz,1H) 6.60(S,2H) As shown in the examples, the present invention enables efficient manufacture of optically active (-)-2-halo-1-(substituted phenyl) ethanol.

Claims: A method of manufacturing (-)-2-halo-1-(substituted phenyl) ethanol which comprises the steps of: bringing a 2-halo-1-(substituted phenyl) ethanone represented by a general formula [ 1 ] (where X represents a chlorine atom or a bromine atom and each of substitution groups R 1 , R 2 , R 3 represents a hydrogen atom, a chlorine atom, a fluorine atom, a methyl group or a methoxy group except cases where 3 substitution groups are all hydrogen atoms) into contact with a microorganism selected from those belonging to Ashbya genus, Brettanomyces genus, Cryptococcus genus, Pichia genus, Rhodosporidium genus, and Trigonopsis genus capable of reducing it asymmetrically into (-)-2-halo-1-(substituted phenyl) ethanol represented by a general formula [ 2 ] (where X and substitution groups R 1 , R 2 , R 3 are the same as in the general formula [ 1 ] and * indicates an asymmetric carbon atom), and collecting (-)-2-halo-1-(substituted phenyl) ethanol. A method of manufacturing (-)-2-halo-1-(substituted phenyl) ethanol which comprises the steps of: bringing a 2-halo-1-(substituted phenyl) ethanone represented by a general formula [ 1 ] (where X represents a chlorine atom or a bromine atom and each of substitution groups R 1 , R 2 , R 3 represents a hydrogen atom, a chlorine atom, a fluorine atom, a methyl group or a methoxy group except cases where 3 substitution groups are all hydrogen atoms) into contact with a microorganism selected from those belonging to Ashbya gossypii, Brettanomyces custersianus, Candida intermedia, Candida krusei, Candida magnoliae, Candida pinus, Candida saitoana, Candida tropicalis, Cryptococcus albidus, Cryptococcus terreus, Geotrichum hirtum, Geotrichum loubieri, Pichia farinosa, Pichia membranaefaciens, Rhodosporidium toruloides, Rhodotorula graminis, Rhodotorula minuta, Rhodotorula rubra, Saccharomyces cerevisiae and Trigonopsis variabilis capable of reducing it asymmetrically into (-)-2-halo-1-(substituted phenyl) ethanol represented by a general formula [ 2 ] (where X and substitution groups R 1 , R 2 , R 3 are the same as in the general formula [ 1 ] and * indicates an asymmetric carbon atom), and collecting (-)-2-halo-1-(substituted phenyl) ethanol. A method according to claim 2 wherein the microorganism is selected from those belonging to Ashbya gossypii IFO 0560, Brettanomyces custersianus IFO 1585, Candida intermedia IFO 0761, Candida krusei IFO 0011, Candida magnoliae IFO 0705, Candida pinus IFO 0741, Candida saitoana IFO 0768, Candida tropicalis IFO 1403, Cryptococcus albidus IFO 0378, Cryptococcus terreus IFO 0727, Geotrichum hirtum CBS 189,53, Geotrichum loubieri CBS 252,61, Pichia farinosa IFO 0574, Pichia membranaefaciens IFO 0460, Rhodosporidium toruloides IFO 0871, Rhodotorula graminis IFO 0190, Rhodotorula minuta IFO 0387, Rhodotorula rubra IFO 0383, Saccharomyces cerevisiae IFO 0614 and Trigonopsis variabilis IFO 0671.

Inventors: HASEGAWA JUNZO, KONISHI YUKO, MAEMOTO SHUNICHI, SAWA IKUO, HASEGAWA, JUNZO, KONISHI, YUKO, MAEMOTO, SHUNICHI, SAWA, IKUO, HASEGAWA, JUNZO, 13-4, TAKAOKA 2-CHOME OHKUBO-CHO, KONISHI, YUKO, 32-305, NAKASHINMACHI 1-CHOME, MAEMOTO, SHUNICHI, 63, OKIHAMAMACHI 2-CHOME, SAWA, IKUO, 3-23, HAMADA-CHO 1-CHOME TAKASAGO-CHO
Applicants: KANEGAFUCHI CHEMICAL IND, KANEGAFUCHI KAGAKU KOGYO KABUSHIKI KAISHA
Application year: 1991
Main classification: C12P7
IPC codes: C12P7, C07D303