KR20010027211A - Process for the Resolution of Racemic Alcohol Compounds Containing Quaternary Chiral Carbon and Synthesis of Systhane Derivatives - Google Patents

Abstract

The present invention provides a racemate of a quaternary asymmetric carbon-containing alcohol compound suitably designed as an intermediate of formula (1) in the asymmetric synthesis of cystane, an excellent azole compound as a fungicide, in the presence of acyljuge in an organic solvent at 0 to 60 ° C. Splitting into optically pure respective R- and S-isomers via transesterification with lipase or hydrolysis of esters of the alcohols at a temperature of and synthesis of cystane and its derivatives from each split isomer It is about.

<Formula 1>

Wherein X represents hydrogen or halogen, n represents the number of carbons. In the present invention, X represents the number of carbons having a hydroxy group and a compound substituted with chlorine at the carbon-4 position of hydrogen or benzene ring 1, 2, 3 , 4 was synthesized. Synthesized racemic alcohols using a lipase, the compound of n = 1 was completely divided to synthesize the azole compound of the target compound. In order to measure the cognition ability of the used lipases of the quaternary asymmetric carbon, n was increased and synthesized to perform transesterification reaction. In order to improve the cleavage, the cleavage was improved by selecting amines as a reagent to remove aldehydes generated during the reaction.

Description

Process for the Resolution of Racemic Alcohol Compounds Containing Quaternary Chiral Carbon and Synthesis of Systhane Derivatives

The present invention is directed to a method of partitioning racemates of racemic alcohol compounds containing quaternary asymmetric carbons into optically pure single isomers using biocatalysts. More specifically, the present invention provides a transesterification reaction of a racemate of a racemate alcohol compound containing quaternary asymmetric carbon with a lipase at a temperature of 0 to 60 ° C. in the presence of acyljuge in an organic solvent or an ester of the alcohol. And a method for synthesizing cystan and its derivatives from each of the divided isomers by optically pure R- and S-isomers through the hydrolysis reaction.

Many biologically active organic compounds contain asymmetric carbons, which are optically active. When preparing organic compounds containing such asymmetric carbons, their isomers are generally produced in an isomeric mixture in a 50/50 ratio. Among these racemates, it is often the case that one isomer has a significantly higher biological activity and the other isomer is inactive or has a particularly different biological activity. For example, triadimenol can have four isomers, the (-)-(1S, 2R) -isomer is the (+)-(1R, 2R) -isomer and the (-)-(1S, 2S) The) -isomers are more active than the (+)-(1R, 2S) -isomers, respectively. Dichlorobutrazole is known to have high activity of (1R, 2R) -isomer among four isomers. Also, it is known that the compound of etaconazole has a higher bactericidal effect than the other isomers of (+)-(2S, 4S)-and (-)-(2S, 4R) -isomers.

In this way, if only one isomer having a high activity can be selectively manufactured, a high amount can be obtained using a small amount, and thus, there is an advantage of reducing environmental pollution due to the use of chemicals. Especially in the case of pharmaceuticals, it is very important that only one isomer be prepared when one isomer is toxic to the human body.

Asymmetric syntheses for the preparation of such homoisomers have been actively studied in recent years and are generally approached by chemical methods. However, there are cases where a biocatalyst is used to selectively prepare only one isomer of a compound having structural properties that is difficult to asymmetrically synthesize by chemical methods. In particular, if a commercially available and inexpensive biocatalyst is available, it may be an advantage over chemical synthesis.

On the other hand, among various biologically active compounds containing asymmetric carbon atoms, especially fenapanil, Fenbuconazole, or Systane represented by the following chemical formula including quaternary asymmetric carbons have a bactericidal effect. It is widely used to protect crops.

Sterile active compounds containing such quaternary asymmetric carbon are generally prepared through several reaction processes using phenylacetonitrile as starting material. The compounds thus prepared are obtained as a mixture of isomers, which are used as such for sterilization purposes.

However, the results of the selective synthesis and cleavage of isomers for these have not been reported, and there has been no study on the difference in activity between the isomers.

Conventionally, the method of dividing optically pure isomers from the racemates of compounds containing asymmetric carbons using biocatalysts has been mainly applied only to alcohol compounds containing secondary asymmetric carbons, and alcohols containing quaternary asymmetric carbons. Examples applied to the cleavage of compounds have not been reported yet.

In general, as a biocatalyst for dividing optically pure isomers from racemates, lipases or esterases, which are hydrolyases, are commonly used. Representative strains that produce them are Pseudomonas and Aspergillus. , Mukomihai, Kandaru Gosa, Alkali Jenners and the like. Since the hydrolase produced from these are different in their reactivity and reaction specificity, it is very important to find efficient partitioning of the target compound substrate.

It is therefore an object of the present invention to provide a method for cleaving optically pure isomers using enzymes that are biocatalysts from racemic alcohol compounds containing quaternary asymmetric carbons, which are biologically important organic intermediates.

It is another object of the present invention to provide a process for preparing triazole-based biologically active compounds into optically pure monoisomers from the pure isomers obtained according to the cleavage method.

Another object of the present invention is to provide a method for determining the absolute structure of optically pure quaternary asymmetric carbon compounds.

In addition, the results obtained in this way can help to reveal the difference in biological activity among the isomers of the fungicide, and it is thought that the effective crop control will be used by this.

Hereinafter, the present invention will be described in detail.

According to the present invention, racemates of alcohol compounds containing quaternary asymmetric carbons are subjected to transesterification reactions or hydrolysis reactions to esters of the alcohols using lipases in the presence of scavengers and scavengers in organic solvents. Optically pure from racemates of (R, S) -alcohol, including methods of dividing into optically pure respective R- and S-isomers and methods of synthesizing cystane and its derivatives from each of the divided isomers Methods of dividing homoisomers are provided.

In the present invention, the (R, S) -alcohol containing the quaternary asymmetric carbon generally means having a structure represented by the following general formula (1).

Wherein R ₁ is a lower alkyl group such as methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, t-butyl or an ether bond such as methoxymethyl, ethoxymethyl, propyloxymethyl or butyloxymethyl And lower alkoxy-lower alkyl groups. R ₂ generally represents a straight or branched chain alcohol group represented by — (CH ₂ ) _n OH, where n is a number from 1 to 5. X substituted in the phenyl ring contains halogens such as chloride, bromide, fluoride and the like and lower alkyl groups such as methyl, ethyl, propyl, isopropyl, isobutyl, t-butyl, ortho, meta and para positions of the phenyl ring May be substituted at each or two or more positions at the same time.

As used herein, "lower alkyl" and "lower alkoxy" refer to straight or branched chain alkyl and alkoxy having 1 to 4 carbon atoms, respectively.

(R, S) -alcohol containing such quaternary asymmetric carbon may be prepared by the following method.

a) R ₁ Br, DMF, NaH, 0 ^o C → room temperature.

b) (CH ₂ O) n, Br (CH ₂ ) n OTHP, DMF, NaH, 0 ^o C → room temperature.

c) 1N methanolic HCl.

In the above scheme, since it is advantageous to use a halogen derivative in order to introduce the R ₂ group to the alpha position of phenylacetonitrile, it is necessary to use the hydroxyl group required for the cleavage after reacting with a protecting group. In this case, the hydroxy group may be dihydropyran, trialkylsilyl halide, benzyl halide, 4-methoxybenzyl halide, etc., but dihydropyran is particularly preferable (R ₂ = XR-OTHP).

Subsequently, the (R, S) -alcohol racemic body containing the quaternary asymmetric carbon prepared by this method is divided using an enzyme as a biocatalyst. Among these enzymes, lipases generally have the advantage that they can be used simultaneously for hydrolysis or esterification. As a lipase, commercially available enzymes include enzymes produced from Pseudomonas sephacia, Pseudomonas fruolesons, Candida lugosa, Aspezius niger, Mukomihai, etc., but in the present invention, Pseudomonas sephasia, Lipases produced from Pseudomonas Furuoroses and Candida Lugosa have the best splitting effects.

The enzymatic reaction according to the method of the present invention is carried out in an organic solvent, because the (R, S) -alcohol racemate, which is the substrate to be subjected to the enzymatic reaction, is well dissolved in the organic solvent. Organic solvents that can be used in the process of the present invention are non-polar, such as n-pentane, n-hexane, n-heptane, cyclohexane, benzene, toluene, xylene, acetonitrile, ethyl acetate, dichloromethane, tetrahydrofuran, dimethylform Polar solvents such as aldehydes and dimethyl sulfoxide may be used, but nonpolar solvents are advantageous. If the reactant is in excess, a small amount of polar solvent may be added.

The reaction temperature was possible at 0-60 ^o C but the best results were obtained at 30-35 ^o C.

According to the present invention, the efficiency of the splitting according to the transesterification reaction can be increased by controlling the reaction temperature and the solvent used in the reaction.

In addition, in the present invention, only a reaction for advancing (R, S) -alcohol to an ester is required, but an acyljuge should be used to prevent a reverse reaction because the enzyme participates in a reversible reaction. As the acyljuge used, vinyl acetate, acetyl anhydride, isopropenyl acetate and the like can be used, but vinyl acetate is most effective. This is because formaldehyde produced after the reaction is easily released.

Aldehydes, which are side reactions generated from the vinyl acetate used as acyl alcohol in the method of the present invention, may act deadly on the enzyme, thereby reducing the reactivity of the enzyme or reducing the reaction specificity. Therefore, there is a need for an aldehyde scavenger capable of removing the generated aldehydes. As the aldehyde scavenger, amines such as pyridine and triethylamine, potassium carbonate, sodium carbonate and the like can be used. In the present invention, when using pyridine and potassium carbonate, it showed the best reaction rate and reaction specificity.

In addition, the method of the present invention may be repeated two to three times the enzyme reaction to increase the splitting effect.

The absolute structure of each isomer of (R, S) -alcohol racemate (I-1) containing quaternary asymmetric carbon prepared by the method of the present invention is crystallized by synthesizing diastereomer having a structure represented by the following formula It can be seen from the X-ray structural crystallography of one diaisomer (VII-1).

a) Jones reagents (sulfuric acid, chromium oxide), acetone,

b) (R) -methyl benzyl amine, DCC, DhbtOH, DMF / CH ₂ Cl ₂

The absolute structure of (R, S) -alcohol racemic compounds, including other quaternary asymmetric carbons, can also be determined by chemical conversion of one isomer determined from the X-ray structural crystallography. For example, the conversion of the carbon chain having a hydroxy group can be prepared by the following method.

Hereinafter, although an Example demonstrates this invention more concretely, this invention is not limited to these Examples.

<Production example 1>

Synthesis of 1-cyano-1-phenylpentane (Formula IV)

In a round bottom three neck flask (250 mL), sodium 60 hydride (2.26 g, 56.4 mmol) in oil is suspended in dried dimethylformaldehyde (70 mL). A solution of phenylacetonitrile (6.06 g, 51.7 mmol) in dimethylformaldehyde (10 mL) at 0 ^° C. was slowly added dropwise. After stirring for 30 minutes, benzyl bromide (6.44 g, 47.0 mmol) was added dropwise at a temperature of 15 ^° C. and stirred for 15 hours. After all the starting materials reacted, the reaction was terminated by pouring into iced water and extracted three times with 50 mL of ethyl ether. The combined solution was washed with saturated sodium bicarbonate, brine and distilled water, dried over anhydrous magnesium sulfate, and the solvent was removed under reduced pressure to obtain 14 g of the crude product title compound. The crude product was separated by silica gel column chromatography (n-hexane / ethyl acetate, 10: 1) to give 6.48 g (80) of the pure compound. Thin layer chromatography retention value 0.33 (n-hexane / ethyl acetate, 9: 1); Gas Chromatography / Mass Spectrometry Analysis Retention Time 7.24 minutes, m / e 51, 57, 65, 77, 89, 103, 117 (100), 130, 145, 158, 173 (M ⁺ ); ¹ H-NMR (250 MHz, CDCl ₃ ) δ 0.90 (t, J = 8.5 Hz, 3H), 1.33-1.47 (m, 4H), 1.86-1.97 (m, 2H), 3.76 (dd, J = 8.0 Hz and J '= 10.1 Hz, 1H), 7.28-7.40 (m, 5H, benzene CH); ¹³ C-NMR (63 MHz, CDCl ₃ ) δ 14.18, 22.50, 29.53, 36.04, 37.81, 121.37, 127.65, 128.39, 129.45, 136.49

<Production example 2>

Synthesis of 2-cyano-2-phenyl-1-hexanol (Formula V, X = H, R ₁ = butyl, R ₂ = CH ₂ OH)

Sodium hydride (1.2 g, 30 mmol, 60 oil dispersion) was dispersed in well-dried dimethylformamide (50 mL) in a round bottom three neck flask (250 mL). Subsequently, the solution of 1-cyano-1-phenylpentane (4.33 g, 25 mmol) prepared in Preparation Example 1 above was slowly added dropwise to 5 mL of dimethylformamide at 0 ^° C. After stirring for 30 minutes, the suspension was kept at 15 ^° C. and paraformaldehyde (1.50 g) in solid form was added in portions, followed by stirring for 15 hours. After all the starting materials had reacted, the mixture was poured slowly into ice water (100 mL) to terminate the reaction, and extracted three times with ethyl ether (50 mL). The extracted organic solvent layer was washed with saturated sodium carbonate solution, brine and distilled water, dried over anhydrous magnesium sulfate, and then the solvent was removed under reduced pressure to obtain 5.5 g of crude product 2-cyano-2-phenyl-1-hexanol. The crude product was separated by silica gel column chromatography (n-hexane / ethyl acetate, 4: 1) to give 4.37 g (86) of the pure title compound. Thin layer chromatography retention value 0.56 (n-hexane / ethyl acetate, 2: 1); Gas Chromatography / Mass Spectrometry Analysis Retention Time 8.80 minutes, m / e 51, 63, 77, 91, 103, 117, 130, 145, 158, 173, (100), 184 (M ⁺ -18); ¹ H-NMR (250 MHz, CDCl ₃ ) δ 0.86 (t, J = 8.6 Hz, 3H), 1.10-1.59 (n, 4H), 180-215 (m, 2H), 1.91 (br. S, 1H,- OH), 3.89 (s, 2H. -CH _2- ), 7.34-7.46 (m, 5H, benzene, CH); ¹³ C-NMR (63 MHz, CDCl ₃ ) δ 14.17, 23.02, 27.44, 35.70, 51.54, 69.83, 122.00, 126.80, 128.69, 129.55, 136.25

<Production example 3>

Synthesis of 2-cyano-2-phenyl-1-hexyl acetate (Formula V, X = H, R ₁ = butyl, R ₂ = CH ₂ OCOCH ₃ )

In a round bottom three neck flask (250 mL), 2-cyano-2-phenyl-1-hexanol (4.4 g, 21.65 mmol) synthesized in Preparation Example 3 was dissolved in dried dichloromethane (20 ml), and a catalyst amount of 4, 4-Dimethylaminopyridine and excess pyridine (5 ml) and acetic anhydride (10 ml) were added. The mixed solution was stirred at room temperature for 2 hours, poured into 10 aqueous hydrochloric acid solution (30 ml) to complete the reaction, and then extracted three times with ethyl ether (30 mL). The extracted organic solvent layer was washed with saturated sodium carbonate solution, brine and distilled water, dried over anhydrous magnesium sulfate, and then the solvent was removed under reduced pressure to obtain 5.1 g of crude product 2-cyano-2-phenyl-1-hexyl acetate. The crude product was separated by silica gel column chromatography (n-hexane / ethyl acetate, 10: 1) to give 4.94 g (93) of the pure title compound. Thin layer chromatography retention value 0.38 (n-hexane / ethyl acetate, 4: 1); Gas Chromatography / Mass Spectrometry Analysis Retention Time 9.23 min, m / e 51, 63, 77, 91, 103, 166, 130, 145, 158, 173 (100), 186, 215, 245 (M ⁺ ); ¹ H-NMR (250 MHz, CDCl ₃ ) δ 0.86 (t, J = 8.1 Hz, 3H), 1.12-1.50 (m, 4H), 1.8-1.95 (m, 2H), 2.06 (s, 3H, OCOCH ₃ ) , 4.36 (s, 2H, -CH ₂ 0-), 7.33-7.45 (m, 5H, benzene CH); ¹³ C-NMR (63 MHz, CDCl ₃ ) δ 13.52, 20.35, 22.28, 26.66, 35.88, 47.85, 68.36, 120.45, 126.01, 128.18, 128.59, 135.19, 169.94

<Production example 4>

Synthesis of Absolute (R) -N- (α-Methylbenzyl)-(S) -2-butyl-2-cyano-2-phenylacetamide (Formula VII) and Determination of Absolute Structure

(1) 0.80 g (3.94 mmol) of 2-cyano-2-phenyl-1-hexanol, a racemic mixture synthesized in Preparation Example 2, was dissolved in a round bottom flask (50 mL) in 10 ml of acetone, and After adding 6 mL of Johnson reagent, the mixture was stirred at room temperature for 3 hours. The reaction was poured into 20 mL of cold water to terminate the reaction and extracted twice with 30 mL of ethyl ether. The extracted organic solvent layer was washed several times with plenty of water, dried over anhydrous magnesium sulfate, and then the solvent was removed under reduced pressure to obtain 710 mg of an intermediate product (± -2-cyano-2-phenylhexanic acid), which was used for the next reaction. [Gas chromatography / mass spectrometer detector analysis retention time 4.15 minutes m / e 57, 63, 77, 89, 103, 117 (100), 130, 144, 156, 173 {M ⁺ -44 (carbon dioxide)}]

(2) 300 mg (1.38 mmol) of the racemic mixture (± -2-cyano-2-phenylhexaoic acid) synthesized before the other round bottom three-neck flask (25 mL) was dried with 3 mL of dried dichloromethane and 3 mL of dimethylformaldehyde. After dissolving in a mixed solution, 270 mg (1.66 mmol) of 3-hydroxy-1,2,3-benzotriazine-4 [3H] -one and 341 mg (1.66 mmol) of dicyclohexylcarbodiimide (DCC) at 0 ° C The mixture was stirred for 30 minutes, and 201 mg (0.21 ml, 1.66 mmol) of (R)-(+)-methylbenzylamine was added to the mixture at the same temperature, and the mixture was stirred at room temperature for 3 hours. After all reactions, 20 ml of ethyl acetate was poured and stirred for 30 more minutes. The white filtrate was removed using a Buchner funnel, washed with saturated sodium carbonate solution, brine and distilled water, dried over anhydrous magnesium sulfate, and then the solvent was removed under reduced pressure. 2-Cyano-2-phenylhexanoxy 600 mg (1-phenylethyl) amide were obtained, the crude product was separated by silica gel column chromatography (n-hexane / ethyl acetate, 10: 1) to give 380 mg (86) of pure diastereomer mixture; mp 83-85 Crystallization and recrystallization with methanol gave 70 mg of one isomer, and the X-ray diffraction structure crystal showed that it was (7R, 10S) -isomer.

(7R, 10S) -isomer: mp: 100-103 ° C., thin layer chromatography retention value 0.49 (n-hexane / ethyl acetate, 10: 1); Gas chromatography / mass spectrometry detector analysis; Retention time 12.19 min m / e 51, 77, 91, 105 (100), 130, 145, 173, 190, 201, 215, 248, 277, 305, 320 (M ⁺ ); [a] _D ²⁸ +23.21 (c = 1.77, Chloroform); ¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.84 (t, J = 3.6 Hz, 3H, -C (21) H ₃ ), 1.25 (m, 4H, C (19) H ₂ , C (20) H ₂ ), 1.37 (d, J = 11.1 Hz, 3H, C (8) H ₃ ), 2.02-2.08 (m, 1H), 2.34-3.40 (m, 1H), 5.03-5.11 (m, 1H, chiral C ( 7) H), 6.38 (d, J = 7.4 Hz, 1H, N (2) H), 7.26-7.60 (m, 10H, benzene CH); ¹³ C-NMR (75 MHz, CDCl ₃ ) δ14.16, 21.91, 22.11, 28.07, 38.33, 50.50, 54.88, 120.75, 126.32, 126.46, 128.04, 129.12, 129.21, 129.53, 135.97, 142.71, 165.90; IR (neat, cm ^-1 ) 3370 (amide NH), 3150 (benzene CH), 2964 (aliphatic CH), 2932 (aliphatic CH), 2868 (aliphatic CH), 2238 (CN), 1686 (C = O), 1526, 1450, 1250, 1025, 764, 738, 700, 592

(7R, 10R) -isomer: thin layer chromatography retention value: 0.52 (n-hexane / ethyl acetate, 10: 1); Gas chromatography / mass spectrometry detector analysis; Retention time 12.08 min m / e 51, 77, 91, 105 (100), 130, 145, 173, 190, 201, 215, 249, 264, 277, 305, 320 (M ⁺ ); ¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.90 (t, J = 3.6 Hz, 3H, -C (21) H ₃ ), 1.25 (m, 4H, C (19) H ₂ , C (20) H ₂ ), 1.37 (d, J = 11.1 Hz, 3H, -C (8) H ₃ ), 2.06 (m, 1H), 2.38 (m, 1H), 5.04 (m, 1H, chiral C (7) H), 6.35 (d, J = 7.4 Hz, 1H, N (2) H), 7.26-7.60 (m, 10H, benzene CH); ¹³ C-NMR (75 MHz, CDCl ₃ ) δ14.16, 22.11, 22.78, 28.07, 37.99, 50.45, 54.80, 120.75, 126.16, 126.41, 127.87, 129.03, 129.21, 129.53, 135.88, 142.52, 165.77

<Production example 5>

Synthesis of 3-cyano-3-phenyl-1-heptanol (Formula V, X = H, R ₁ = butyl, R ₂ = CH ₂ CH ₂ OH)

(1) In a round bottom three neck flask (250 mL), sodium hydride (460 mg, 11.5 mmol, 60 oil dispersion) was dispersed in well-dried dimethylformamide (50 mL). A solution of 1-cyano-1-phenylpentane (1.66 g, 9.6 mmol) prepared in Preparation Example 1 at 0 ^° C. was slowly added dropwise to 5 mL of dimethylformamide. After stirring for 30 minutes, 2-tetrahydropyranyloxyethyl bromide [2.0 g, 9.6 mmol, using a bromine ethanol as a starting material to give a general hydroxyl group protection reaction at a temperature of 15 ^° C. Can be synthesized. The synthesis method is as follows. In a round bottom three neck flask (250 mL), 2-bromoethanol (2.49 g, 20 mmol), 3,4-dihydro-2H-pyran (2.02 g, 2.19 ml, 24 mmol) and a catalytic amount (about 100 mg) of pyridinium para- Toluene sulfone was dissolved in 50 ml of dried dichloromethane and stirred at room temperature for 3 hours. Ethyl ether (50 ml) was poured and diluted, washed with half saturated brine to remove the catalyst, dried over anhydrous magnesium sulfate, and then the solvent was removed under reduced pressure to obtain 4.30 g of crude product. The crude product was separated by silica gel column chromatography (n-hexane / ethyl acetate, 20: 1) to give 3.97 g (95) of the pure compound. Gas Chromatography / Mass Spectrometry Analysis Retention Time 5.61 min, m / e 56, 73, 85 (100), 93, 109, 115, 135, 153, 163, 180, 209 (M ⁺ )] Stirred for time. After all the starting materials had reacted, the reaction was terminated by pouring into iced water and extracted three times with 50 mL of ethyl ether. The extracted organic solvent layer was washed with saturated sodium carbonate solution, brine and distilled water, dried over anhydrous magnesium sulfate, and then the solvent was removed under reduced pressure to obtain the crude product 3-cyano-3-phenyl-1-heptanol dihydropyranyl ether ( 3.70 g of Formula V, X = H, R ₁ = butyl, R ₂ = CH ₂ CH ₂ OTHP) were obtained. This compound was not a stable compound and was used directly in the next reaction.

(2) In a round bottom flask (50 mL), the crude product 3-cyano-3-phenyl-1-heptanol dihydropyranyl ether (3.70 g) obtained in (1) was dissolved in methanol (20 ml), 1N hydrochloric acid (5 ml) was added under methanol solvent, and the mixture was stirred at room temperature for 12 hours. Methanol was removed under reduced pressure and extracted three times with ethyl ether (50 mL) by addition of saturated sodium carbonate solution. The extracted organic solvent layer was washed with saturated sodium carbonate solution, brine and distilled water, dried over anhydrous magnesium sulfate, and then the solvent was removed under reduced pressure to obtain 3.8 g of a crude product. The crude product was separated by silica gel column chromatography (n-hexane / ethyl acetate, 4: 1) to give 1.73 g (83) of pure compound. Thin layer chromatography retention value 0.34 (n-hexane / ethyl acetate, 4: 1); Gas chromatography / mass spectrometry analysis retention time 9.42 min; m / e 51, 57, 77, 91, 103, 116, 130, 143 (100), 161, 173, 186, 199, 217 (M ⁺ ); ¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.85 (t, J = 7.3 Hz, 3H), 1.03-1.21 (m, 2H), 1.20-1.42 (m, 2H), 1.49 (br, s, 1H, − OH), 1.60-1.75 (m, 1H), 1.82-2.17 (m, 3H), 3.58 (t, J = 6.2 Hz, 2H, -CH ₂ O-), 7.30-7.40 (m, 5H, benzene CH)

<Production example 6>

Synthesis of 3-cyano-3-phenyl-1-heptyl acetate (Formula V, X = H, R ₁ = butyl, R ₂ = CH ₂ CH ₂ OCOCH ₃ )

3-Cyano-3-phenyl-1-heptyl acetate was prepared from the 3-cyano-3-phenyl-1-heptanol prepared in Preparation Example 5 as starting materials, under the same conditions as the synthesis method of Preparation Example 3. Prepared.

Yield 94, thin layer chromatography retention value 0.46 (n-hexane / ethyl acetate, 2: 1); Gas Chromatography / Mass Spectrometry Analysis Retention Time 9.77 min, m / e 51, 73, 75, 101, 115, 137, 150, 164, 180, 207 (100), 209, 249, 279 (M ⁺ ); ¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.88 (t, J = 8.2 Hz, 3H), 1.10-1.48 (m, 4H), 1.82-1.93 (m, 4H), 2.11 (s, 3H, OCOCH ₃ ) , 4.30-4.37 (m, 2H, -CH ₂ O-), 7.33-7.45 (m, 5H, benzene CH)

<Production example 7>

Synthesis of 4-cyano-4-phenyl-1-octanol (Formula V, X = H, R ₁ = butyl, R ₂ = CH ₂ CH ₂ CH ₂ OH)

(1) 3-tetrahydropyranyloxypropyl bromide [2.1 g, 9.6 mmol, using only 1-cyano-1-phenylpentane and an alkylating reagent under the same conditions as in Synthesis in Preparation Example 5; 3-bromopropanol can be used as a starting material for general hydroxyl protection reactions. The synthesis method is as follows. 6.95 g (50 mmol) of 3-bromopropanol, 5.04 g (5.47 ml, 60 mmol) of 3,4-dihydro-2H-pyran and a catalytic amount (about 200 mg) of pyridinenium para-toluene sulfone in a round bottom three neck flask (250 mL) Was dissolved in 50 ml of dried dichloromethane and stirred at room temperature for 3 hours. 50 ml of ethyl ether was poured and diluted, washed with half saturated brine to remove the catalyst, dried over anhydrous magnesium sulfate, and then the solvent was removed under reduced pressure to obtain 12.0 g of crude product. The crude product was separated by silica gel column chromatography (n-hexane / ethyl acetate, 20: 1) to give 10.5 g (94) of the pure compound. Thin layer chromatography retention value of 0.25 (n-hexane / ethyl acetate, 20: 1); Gas Chromatography / Mass Spectrometry Analysis Retention Time 6.59 min; m / e 56, 67, 85 (100), 93, 107, 123, 137, 151, 167, 196, 223 (M ⁺ ); ¹ H-NMR (300 MHz, CDCl ₃ ) δ 1.15-1.63 (m, 6H), 2.02-2.10 (m, 2H), 3.43-3.48 (m, 4H, -OCH _{2 ×} 2), 3.76-3.84 (m , 2H); 4.53 (t, J = 3.4 Hz, 1H, chiral CH); ¹³ C-NMR (75 MHz, CDCl ₃ ) δ19.8, 25.7, 30.9, 30.9, 33.2, 62.5, 65.1, 99.1], 4-cyano-4-phenyl-1-octanol dihydro pyranyl ether Was prepared. Likewise, this compound was not a stable compound and was used directly in the next reaction.

(2) Starting material under the same conditions as in Synthesis Example 5, crude product 4-cyano-4-phenyl-1-octanol dihydropyranyl ether synthesized in (1) was used. No-4-phenyl-1-octanol was obtained.

Yield 80; Thin layer chromatography retention value 0.38 (n-hexane / ethyl acetate, 2: 1); Gas Chromatography / Mass Spectrometry Analysis Retention Time 9.62 min; m / e 51, 57, 77, 91, 103, 116, 129 (100), 142, 156, 172, 189, 204, 231 (M ⁺ ); ¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.84 (t, J = 7.3 Hz, 3H), 1.03-1.20 (m, 1H), 1.22-1.48 (m, 4H), 1.50 (br, s, 1H,- OH), 1.63-1.78 (m, 1H), 1.84-2.17 (m, 4H), 3.59 (t, J = 6.18 Hz, 2H, -CH ₂ O-), 7.30-7.40 (m, 5H, benzene CH)

<Production example 8>

Synthesis of 4-cyano-4-phenyl-1-octyl acetate (Formula V, X = H, R ₁ = butyl, R ₂ = CH ₂ CH ₂ CH ₂ OCOCH ₃ )

4-Cyano-4-phenyl-1-octyl acetate was prepared from the 4-cyano-4-phenyl-1-octanol prepared in Preparation Example 7 as starting materials under the same conditions as the synthesis method described in Preparation Example 3. Prepared.

Yield 92, thin layer chromatography retention value 0.45 (n-hexane / ethyl acetate, 2: 1); Gas Chromatography / Mass Spectrometry Analysis Retention Time 10.39 min, m / e 57, 73, 77, 101 (100), 116, 129, 145, 156, 175, 200, 213, 230, 246, 273 (M ⁺ ) ; ¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.86 (t, J = 8.2 Hz, 3H), 1.05-1.43 (m, 4H), 1.58-1.70 (m, 2H), 1.82-1.93 (m, 4H), 2.11 (s, 3H, OCOCH ₃ ), 4.30-4.37 (m, 2H, -CH ₂ O-), 7.33-7.45 (m, 5H, benzene CH)

<Production example 9>

Synthesis of 5-cyano-5-phenyl-1-nonanol (Formula V, X = H, R ₁ = butyl, R ₂ = CH ₂ CH ₂ CH ₂ CH ₂ OH)

(1) 4-tetrahydropyranyloxybutyl chloride [2.1 g, 9.6 mmol, using the 1-cyano-1-phenylpentane and alkylation reagent only as starting materials under the same conditions as in Synthesis in Preparation Example 5; Using 4-chlorobutanol as a starting material can be synthesized by a general hydroxyl group protection reaction. The synthesis method is as follows. 4-chlorobutanol (3.26 g, 3.5 ml, 30 mmol), 3,4-dihydro-2H-pyran (3.91 g, 3.59 ml, 36 mmol) and a catalytic amount (about 100 mg) of pyridinium in a round bottom three neck flask (100 mL) Para-toluene sulfone was dissolved in 25 ml of dried dichloromethane and stirred at room temperature for 3 hours. 50 ml of ethyl ether was poured and diluted, washed with half saturated brine to remove the catalyst, dried over anhydrous magnesium sulfate, and then the solvent was removed under reduced pressure to obtain 6.2 g of crude product. The crude product was separated by silica gel column chromatography (n-hexane / ethyl acetate, 20: 1) to give 5.66 g (98) of the pure compound. Thin layer chromatography retention value 0.28 (n-hexane / ethyl acetate, 20: 1); Gas Chromatography / Mass Spectrometry Analysis Retention Time 6.60 minutes; m / e 55, 67, 85, (100), 93, 101, 115, 134, 157, 191, 193 (M ⁺ ); ¹ H-NMR (300 MHz, CDCl ₃ ) δ 1.45-1.62 (m, 4H), 1.70-1.79 (m, 4H), 1.80-1.90 (m, 2H), 3.35-3.41 (m, 1H), 3.42- 3-cyano- using 3.49 (m, 1H), 3.58 (t, J = 6.6 Hz, 2H, -CH ₂ Cl), 3.75-3.84 (m, 2H), 4.50-4.59 (m, 1H)]. 5-phenyl-1-nonanol dihydro pyranyl ether was prepared. Likewise, this compound was not a stable compound and was used directly in the next reaction.

(2) Starting material under the same conditions as in Synthesis Example 5, crude product 5-cyano-5-phenyl-1-nonanol dihydro pyranyl ether synthesized in (1) above was used. No-5-phenyl-1-nonanol was obtained.

Yield 89; Thin layer chromatography retention value 0.29 (n-hexane / ethyl acetate, 1: 1); Gas chromatography / mass spectrometer analysis retention time 10.42 min; m / e 55, 65, 77, 91, 103, 116, 129, 143, 158, 173 (100), 189, 202, 218, 227, 245 (M ⁺ ); ¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.84 (t, J = 7.3 Hz, 3H), 1.01-1.38 (m, 5H), 1.39-1.60 (m, 3H), 1.50 (br, s, 1H, − OH), 1.81-2.09 (m, 4H), 3.58 (t, J = 6.2 Hz, 2H, -CH ₂ O-), 7.27-7.39 (m, 5H, benzene CH)

<Production example 10>

Synthesis of 5-cyano-5-phenyl-1-nonyl acetate (Formula V, X = H, R ₁ = butyl, R ₂ = CH ₂ CH ₂ CH ₂ CH ₂ OCOCH ₃ )

5-Cyano-5-phenyl-1-nonyl acetate was prepared from the 5-cyano-5-phenyl-1-nonanol prepared in Preparation Example 9 as starting materials under the same conditions as the synthesis method of Preparation 3 Prepared.

Yield 95; Thin layer chromatography retention value of 0.52 (n-hexane / ethyl acetate, 2: 1); Gas Chromatography / Mass Spectrometry Analysis Retention Time 10.90 min, m / e 55, 61, 91, 103, 115, 123 (100), 145, 173, 189, 200, 227, 244, 260, 287 (M ⁺ ) ; ¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.86 (t, J = 8.2 Hz, 3H), 1.08-1.37 (m, 4H), 1.54-1.73 (m, 4H), 1.82-1.93 (m, 4H), 2.13 (s, 3H, OCOCH ₃ ), 4.28-4.36 (m, 2H, -CH ₂ O-), 7.27-7.40 (m, 5H, benzene CH)

<Production example 11>

Synthesis of 2-phenyl-2- (1H-1,2,4-triazol-1-ylmethyl) hexanenitrile: (R) -isomer

(1) In a round bottom three neck flask (25 mL), (R) -2-cyano-2-phenyl-1-hexanol (0.10 g, 0.5 mmol) and triethylamine (148 mg, 0.21 mL, 1.47 mmol) were diluted with Dissolved in methane (5 ml). Methanesulfonylchloride (68 mg, 0.5 mmol) was dissolved in dichloromethane (1 ml) and slowly added dropwise at 0 ^° C. After stirring for 15 hours, the mixture was poured into iced water and extracted three times with ethyl ether (50 mL). The combined solution was washed with dilute aqueous hydrochloric acid solution, saturated sodium carbonate solution and distilled water, dried over anhydrous magnesium sulfate, and then the solvent was removed under reduced pressure to obtain 0.19 g of a sulfonated crude product. The crude product was separated by silica gel column chromatography (n-hexane / ethyl acetate, 4: 1) to give 131 mg (95) of pure compound. Thin layer chromatography retention value 0.61 (benzene / ethyl acetate, 5: 1); Gas Chromatography / Mass Spectrometry Analysis Retention Time 10.91 min; m / e 51, 65, 79, 103, 116, 129, 145, 172 (100), 185, 202, 224, 238, 251, 281 (M ⁺ ); ¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.83 (t, J = 7.3 Hz, 3H), 1.20-1.55 (m, 4H), 1.87-2.17 (m, 2H), 2.91 (s, 3H, OSO ₂ CH ₃ ), 4.43 (s, 2H, CH ₂ O), 7.24-7.47 (m, 5H, benzene CH); ¹³ C-NMR (75 MHz, CDCl ₃ ) δ 14.11, 22.81, 27.18, 35.94, 38.11, 48.79, 73.23, 120.34, 126.71, 129.29, 129.72, 134.62

(2) 1,2,4-triazole sodium salt (50 mg, 0.55 mmol) in sulfonate compound (131 mg, 0.47 mmol) prepared in (1) above and dimethyl sulfoxide (5 mL) in a round bottom three neck flask (25 mL). ) Was added and stirred at 100 ^° C. for 24 hours. After all the starting materials reacted, the reaction was terminated by pouring into iced water and extracted three times with 20 mL of ethyl ether. The combined solutions were washed with saturated sodium carbonate solution, brine and distilled water, dried over anhydrous magnesium sulfate, and then the solvent was removed under reduced pressure. The residue was purified purely by column chromatography to give (R) -isomer 2-phenyl-2- (1H- (1,2,4-triazol-1-ylmethyl) hexanenitrile (104 mg, 90). (R) -isomer: thin layer chromatography retention value 0.28 (n-hexane / ethyl acetate, 1: 1); [α] _D ²⁷ = + 54.18 (c 0.55, methanol); gas chromatography / mass spectrometry Retention time 10.71 min; m / e 55, 63, 82, 91, 103, 116, 145 (100), 172, 185, 195, 211, 225, 239, 254 (M ⁺ ); ¹ H-NMR (300MHz, CDCl ₃ ) δ 0.83 (t, J = 7.2 Hz, 3H), 1.28-1.53 (m, 4H), 1.84-2.23 (m, 2H), 4.58 (dd, J = 14.2 Hz and J '= 39.3 Hz, 2H , -CH ₂ -triazole), 7.28-7.42 (m, 5H, benzene CH), 7.77 (s, 1H, triazole CH), 7.89 (s, 1H, triazole CH)

<Production example 12>

Synthesis of 2-phenyl-2- (1H- (1,2,4-triazol-1-ylmethyl) hexanenitrile: (S) -isomer

Synthesis was carried out in the same manner as in the preparation procedure in Preparation Example 11, and the 2-phenyl-2- (1H- (1,2,4-triazol-1-ylmethyl) hexanenitrile of the (S) -isomer was 96 in yield. Got it.

(S) -isomer: [α] _D ²⁷ = -61.4 (c 0.55, methanol)

<Example 1>

Split-Transesterification Reaction of 2-cyano-2-phenyl-1-hexanol

Four round bottom three-necked flasks (25 mL) equipped with a thermometer were dissolved in 224 mg (0.11 mmol) of 2-cyano-2-phenyl-1-hexanol in 5 mL of well-dried n-hexane. In each solution, Candida Lugosa (CRL), Pseudomonas Sephacia (LPS), Pseudomonas Furuole Lessons (PFL) A (made by Aldrich), and Pseudomonas Furuole Lessons (LAK) B (made by Amanosa), respectively After the addition of 100, 100, 50 and 50 of each enzyme, 95 mg (0.10 mL, 0.11 mmol) of vinyl acetate was added thereto, and the reaction was confirmed by TLC while stirring at 32-34 ^° C. for a certain time. The results of the reaction are summarized in Table 1.

Conditions and Results Enzymes ¹ Response time (h) Switch ² () ee ( ³ ) alcohol acetate E ⁴ Candidaru Gosa (CRL) One 48 26 (S) 62 (R) 5.5 Pseudomonas Sepia (LPS) 10 11 30 (R) 56 (S) 4.7 Pseudomonas furuo lessons (PFL) A One 20 50 (R) 78 (S) 13 Pseudomonas furuo lessons (LAK) B 24 9 8 (R) 100 (S) 〉 200 48 35 36 (R) 100 (S) 〉 200

1. Candida Lugosa (CRL) Sigma Products: Active (860 units / mg); Pseudomonas Sephacia (LPS) Amanosa Products: Active (> 30,000 units / g); Pseudomonas Furuole Lessons (PFL) A from Aldrich; Pseudomonas Furuoresons (PFL) B Amano's product: Active (> 20,000 units / g)

2. Refers to the percentage of acetate in the absolute area ratio of acetate reacted with unreacted alcohol using a gas chromatography / mass spectrometer. There are no by-products from this enzymatic reaction.

3. The optical purity (enantiomeric excess) is obtained by separating the alcohol and acetate by silica gel column chromatography (n-hexane / ethyl acetate, 10: 1 → 2: 1) after completion of the reaction. It was obtained and measured by chiral column gas chromatography. The analysis conditions are as follows. Column beta-diimide X ^{TM (Beta-DEX TM) 110 (} Fused silica capillary column), column inside diameter (Column ID) 0.25mm, the thickness of the film (Film Thickness) 0.25m, column length (Column Length) 30m, a detector (Detector) Flame ionization detector (FID), detector temperature 250 ° C., injector temperature 250 ° C., analysis program: initial temperature 140 ° C., initial time 40 minutes, temperature control 2 ° C. per minute, final temperature 200 ° C., final time 10 minutes; R-isomer retention time 54.21 minutes, S-isomer retention time 54.88 minutes

The absolute structure of each isomer could be determined by the following method.

The diastereomer obtained by reacting (R)-(+)-methylbenzylamine with the obtained alcohol was determined by comparison with the retention time of the gas chromatography / mass spectrometry detector of the diastereomer in Example 1-4.

4. E (Enantiomeric ratio) is calculated from the following equation. The larger the calculated value, the greater the efficiency of enzyme catalysis.

The above equation is obtained from experimental results and can be calculated from the following equation.

Where ee _s and ee _p are the optical purity of the starting materials and products, and c represents the conversion value.

<Example 2>

Improved partition reaction through repeated product recycling

(R) -isomer (ee 62), (S) -isomer (ee 76) of 2-cyano-2-phenyl-1-hexanol partitioned by Candida Lugosa in Example 6 twice in the same manner The reaction was repeated to obtain isomers with a purity of 98 or more, respectively.

(R) -isomer: [α] _D ²⁵ = + 11.05 (c 0.58, methanol)

(S) -isomer: [α] _D ²⁵ =-11.02 (c 0.58, methanol)

<Example 3>

Split-Transesterification Reaction of 3-cyano-3-phenyl-1-heptanol

In the same manner as in Example 1, 3-cyano-3-phenyl-1-heptanol was transesterified to separate isomers as shown in Table 2 below.

Conditions and Results Response time (h) transform() ee (alcohol acetate) E Candidaru Gosa (CRL) 2 27 16 26 2 Pseudomonas Sephacia (LPS) 52 7 20 100 〉 200 72 23 6 84 12 Pseudomonas furuo lessons (PFL) A 9 39 36 68 7.4

<Example 4>

Split-Transesterification Reaction of 4-cyano-4-phenyl-1-octanol

In the same manner as in Example 1, 4-cyano-4-phenyl-1-octanol was transesterified to separate isomers as shown in Table 3 below.

Conditions and Results Response time (h) transform() ee () ¹ alcohol acetate E Candidaru Gosa (CRL) 2 39 22 (R) 10 (S) 1.5 Pseudomonas Sepia (LPS) 5 47 84 (R) 48 (S) 7.1 Pseudomonas furuo lessons (PFL) A 1.5 98 100 (R) 14 (S) 6.4 0.5 57 97 (R) 40 (S) 8.6

1. The absolute structure of the separated isomer was increased from the (R) -2-cyano-2-phenyl-1-hexanol prepared in Example 1 by increasing the number of carbon atoms of the alkyl group having a hydroxyl group by (R) -4 -Cyano-4-phenyl-1-octanol was determined by comparison with the retention in chiral liquid chromatography. Analytical conditions for chiral liquid chromatography: chiralcel OD column (cellulose carbamate derivative), n-hexane / isopropyl alcohol, 99: 1, effluent 1.5 mL / min, detector UV 254 nm, retention time (R) -isomer 18.80 minutes, (S) -isomer 20.54 minutes

<Example 5>

Split-Transesterification Reaction of 5-cyano-5-phenyl-1-nonanol

In the same manner as in Example 1, 5-cyano-5-phenyl-1-nonanol was transesterified to separate isomers as shown in Table 4 below.

Conditions and Results Response time (h) transform() ee (alcohol acetate) E Candidaru Gosa (CRL) 2 13 20 30 2.2 Pseudomonas Sepia (LPS) 2 72 42 28 2.6

<Example 6>

Split-hydrolysis Reaction of 2-cyano-2-phenyl-1-hexyl Acetate

2-cyano-2-phenyl-1-hexyl acetate (123 mg, 0.50 mmol) prepared in Preparation Example 3 was added to each of the three reactors to which the pH controller was attached, and phosphate buffer solution (8 mL) was added thereto. Candida Lugosa (CRL), Posin Pancreas Lipase (PPL), and Posin River Esterase (PLE) were added to each solution in the same amount by weight, and then the pH was adjusted to 7.0 using a syringe pump. While stirring for a while, the degree of reaction was confirmed by TLC. The reaction results are summarized in Table 5.

Conditions and Results Enzymes ¹ Response time (h) transform() ee (acetate alcohol) E Candidaru Gosa (CRL) 14 56 2 2 1.1 Gun barrel pancreas lipase (PPL) 112 One ^-2 2 - Gunshin River Esterase (PLE) 112 30 12 24 1.8

1. Candida Lugosa (CRL) Sigma Products: Active (860 units / mg); Gunsin Pancreas Lipase (PPL) Sigma; Gunshin River Esterase (PLE) from Furukasa

2. No device analysis

<Example 7>

Transesterification Reaction According to Split-Solvent Change of 2-Cyano-2-phenyl-1-hexanol

In four round bottom three neck flasks equipped with a thermometer (25 mL), 2-cyano-2-phenyl-1-hexanol (224 mg, 0.11 mmol) was dissolved in a well-dried solvent (5 mL). Add the same amount of Candida Lugosa (CRL) to each solution by weight, add vinyl acetate (95mg, 0.10mL, 0.11mmol), and stir at 32-34 ^o C for a certain period of time. It was confirmed. In the transesterification reaction according to the solvent, it was judged that using a non-polar n-hexane as the solvent was the best in the reaction time or the splitting efficiency. The reaction results are summarized in Table 6.

Conditions and Solvents Response time (h) transform() ee () Alcohol (S) Acetate (R) E n-hexane One 48 26 62 5.5 Cyclohexane One 40 13 56 4 benzene 11 47 56 46 4.6 Ethyl acetate 112 15 4 56 3.7 Dichloromethane 112 28 12 48 3.2 Tetrahydrofuran 165 Unreacted -- - Acetonitrile 165 Unreacted -- -

<Example 8>

Transesterification Reaction with Split-Scavenger and 2-Addition of 2-Cyano-2-phenyl-1-hexanol

In four round bottom three-necked flasks (25 mL) equipped with a thermometer, 2-cyano-2-phenyl-1-hexanol (224 mg, 0.11 mmol) was dissolved in n-hexane (5 mL). Add the same amount of Candida Lugosa (CRL) and liquid additives to each solution by weight in each solution, and add 1 volume of solvent and the same amount of reactant, and then add vinyl acetate (95mg, 0.10mL, 0.11mmol), 32-34 ^o The degree of reaction was confirmed by TLC while stirring at a certain time. When pyridine was used as a scavenger in the transesterification reaction according to the additive, the reaction time was judged to be the most excellent in terms of group or splitting efficiency. A summary of the reaction results is shown in Table 7.

Conditions and Result Additives Response time (h) transform() ee () Alcohol (S) Acetate (R) E Pyridine (1) 3.5 21 54 74 11 Triethylamine (1) One 24 20 60 4.8 Diisopropylamine (1) 3 18 32 54 4.5 MoreCure Shiv (4) 15 47 6 56 3.8 Water (1) 20 11 10 54 3.7

According to the present invention, there is provided a process for dividing racemates of racemic alcohol compounds containing quaternary asymmetric carbons into optically pure single isomers using biocatalysts.

Claims (10)

(R, S) -alcohol racemate represented by the following formula is characterized in that it is subjected to esterification or hydrolysis reaction with lipase at 0-60 ° C. in the presence of acyl alcohol and aldehyde scavenger in organic solvent. To separate each of the pure R- and S-isomers from the (R, S) -alcohol racemates of: <Formula 1> In the above formula, R ₁ represents a lower alkyl group or a lower alkoxy-lower alkyl group, R ₂ represents a straight or branched chain alcohol group represented by the formula — (CH ₂ ) _n OH where n is a number from 1 to 5, X represents a halogen or lower alkyl group which may be substituted at the ortho, meta and para positions of the phenyl ring, respectively or at two or more positions at the same time. The method of claim 1, wherein the lipase is selected from the group consisting of Candida Lugosa, Pseudomonas Furuoresons, and enzymes produced from Pseudomonas sefacia. The method of claim 2, wherein the enzyme is used by immobilization on a solid support. The method of claim 1, wherein the acyljuge is selected from the group consisting of vinyl acetate, isopropenyl acetate, acetyl & hydride. The method of claim 1, wherein the organic solvent is normal hexane, cyclo hexane, diisopropyl ether, ethyl acetate, benzene, toluene, tetrahydro hulan, diethyl ether, acetonitrile, dichloroethane, dichloromethane, 1,4- Dioxane, methanol, dimethyl sulfoxide, dimethyl formaldehyde, hexane / ethyl acetate (9/1 -5/5). 2. The aldehyde scavenger according to claim 1, wherein the aldehyde scavenger is a base such as triethyl amine, pyridine, piperidine, piperazine, diisopropylamine, imidazole, calcium carbonate, sodium acetate, and molar curassib (branch), lithium And selected from the group consisting of chromides. The process according to claim 1, wherein the esterification or hydrolysis reaction is carried out twice or three times. Methanesulfonyl chloride of optically pure (R) -2-cyano-2-phenyl-1-hexanol or (S) -2-cyano-2-phenyl-1-hexanol obtained according to the method of claim 1 Or reacted with toluenesulfonyl chloride to prepare a sulfone derivative, Optically pure (R) of 2-phenyl-2- (1H- (1,2,4-triazol-1-ylmethyl) hexanenitrile by reacting the sulfone derivative with 1,2,4-triazole sodium salt Or a process for preparing the (S) -isomer. In order to determine the absolute structure of the optically pure quaternary asymmetric carbon compounds obtained according to the method of claim 1, (R, S) -2-cyano-2-phenyl-1-hexanol is oxidized to carboxylic acid, Reaction with (R)-([alpha])-methylbenzylamine to prepare diastereomers, The diastereomer was crystallized and subjected to X-ray crystallography to determine the (R) -2-cyano-2-phenyl-1-hexanol and (S) -2-cyano-2-phenyl-1-hexanol. How to determine absolute structure. A chemical conversion reaction is carried out from (R) -2-cyano-2-phenyl-1-hexanol and (S) -2-cyano-2-phenyl-1-hexanol whose absolute structure is determined according to the method of claim 9. To determine the absolute structure of 4-cyano-4-phenyl-1-octanol.