JPH05227991A - Production of optically active 3-pyrrolidinol derivative - Google Patents

Abstract

PURPOSE:To obtain an optically active 3-pyrrolidinol useful as an intermediate for pharmaceuticals, etc., on an industrial scale at a low cost by reacting a racemic pyrrolidinol compound with an acylation agent in the presence of an enzyme source having stereoselective acylation activity. CONSTITUTION:The objective optically active 3-pyrrolidinol derivative can be produced by reacting a racemic compound of formula I [R1 is (substituted) benzyl, alkanoyl or aroyl] (e.g. N-benzyl-3-pyrrolidinol) with a compound expressed by the formula R2COOR3 (R2 is alkyl or aryl; R3 is H, alkyl, alkenyl, alkanoyl or aroyl) (e.g. vinyl acetate) in the presence of an enzyme source capable of stereoselectively acylating the compound of formula I (e.g. lipase) to form a compound of formula II (* mark represents that the labeled carbon is optically active) (e.g. N-benzyl-3-acetoxypyrrolidine) and an optically active 3-pyrrolidinol expressed by formula III.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for producing an optically active 3-pyrrolidinol derivative which is an important intermediate of an optically active compound useful as a medicine such as β-lactam antibiotics and dihydropyridine compounds.

[0002]

2. Description of the Related Art Conventionally, methods for producing optically active 3-pyrrolidinol derivatives are roughly classified into (1) a method of synthesizing a chiral compound as a starting material and (2) an asymmetric synthesis using a prochiral compound as a starting material. Alternatively, two methods of synthesizing an optically active substance by resolution are known. Regarding (1), a method of synthesizing from hydroxyproline (JP-A-60-23328) and a method of synthesizing from D-malic acid [Heterocycles, 2
4 , 1331 (1986)], a method of synthesizing from glutamic acid [Synth. Comm (Synth. Comm.
un.) , 16 , 1815 (1986)], a method of synthesizing from aspartic acid [Heterocycles, 24 , 133
1 (1986)], and a method of synthesizing from glutamine [Bulletin of the Chemical Society of Japan (Bull. Chem. Sco. Jpn.) , 51 , 3296 (1978)] and the like. Regarding (2), a method of synthesizing from 4-hydroxy-2-pyrrolidone (JP-A-1-207266) and a method of synthesizing from aminobutanol derivative
(JP-A-3-176462), a method of synthesizing 4-halo-3-hydroxybutanenitrile from a sulfonic acid ester (JP-A-3-176463), and the like are known.

[0003]

SUMMARY OF THE INVENTION The object of the present invention is β-
It is intended to provide a novel method for producing an optically active 3-pyrrolidinol derivative which is a useful intermediate in the synthesis of lactam antibiotics, dihydropyridine compounds and the like.

[0004]

According to the present invention, the formula (II
I)

[0005]

[Chemical 5]

[Wherein R ₁ represents a substituted or unsubstituted benzyl group, alkanoyl group or aroyl group] [hereinafter abbreviated as compound (III)]. ] And the formula (IV)

[0007]

[Chemical 6]

(R ₂ represents an alkyl group or an aryl group, and R ₃ represents a hydrogen atom, an alkyl group, an alkenyl group, an alkanoyl group or an aroyl group.) [Abbreviated as compound (IV) hereinafter. ] In the presence of an enzyme source having an activity of stereoselectively acylating compound (III),

[0009]

[Chemical 7]

(Wherein R ₁ and R ₂ have the same meanings as described above, and * means that the carbon is optically active) and the formula (II).

[0011]

[Chemical 8]

(In the formula, R ₁ has the same meaning as described above.
Indicates that the carbon is optically active. The method for producing an optically active substance of a 3-pyrrolidinol derivative, which comprises obtaining an optically active substance of a 3-pyrrolidinol derivative represented by (4). Hereinafter, the compound represented by the formula (I) is referred to as the compound (I),
The compound represented by I) is referred to as compound (II).

The definition of each group in the formula is as follows.
The substituents of the substituted benzyl are the same or different and are halogen atoms such as chlorine, bromine and iodine having 1 to 3 substituents, lower alkyl groups having 1 to 3 carbon atoms such as methyl, ethyl and propyl, and alkyl moieties having 1 to 1 carbon atoms. And an alkoxy group which is a lower alkyl of 3. As the alkanoyl group, a formyl or alkyl moiety having 1 to 8 carbon atoms is methyl, ethyl, propyl, butyl, pentyl, hexyl,
Examples thereof include a linear alkyl group having 1 to 7 carbon atoms such as heptyl. Examples of the aryl moiety of the aryl group or the aroyl group include phenyl and naphthyl.
Examples of the alkyl group include linear alkyl groups having 1 to 7 carbon atoms such as methyl, ethyl, propyl, butyl, pentyl, hexyl and heptyl.

Examples of the alkenyl group include linear or branched alkenyl having 2 to 8 carbon atoms such as vinyl and isopropenyl. As an enzyme source used in the above-mentioned acylation reaction, either an enzyme having an activity of asymmetrically acylating compound (III) or a substance containing it can be used, either a purified enzyme or a crude enzyme.

Enzymes include triacylglycerol lipase (EC.3.1.1.3, also known as lipase), riboprotein lipase (EC.3.1.1.34), carboxylesterase (E
C.3.1.1.1) is used. Although these enzymes are produced by microorganisms and animal tissues, they are commercially available and can be easily obtained. Specific examples of commercially available enzymes include riboprotein lipase (derived from Pseudomonas genus microorganism; manufactured by Kyowa Medex), Lipase P Amano (derived from Pseudomonas genus microorganism; manufactured by Amano Pharmaceutical Co., Ltd.), polysimpancreatic lipase (derived from porcine pancreas; Sigma company). Manufactured), lipase (derived from Candida genus microorganism; manufactured by Sigma), big river esterase (derived from pig liver; manufactured by Sigma), Neulase F Amano (derived from microorganism belonging to Rhizobus genus; manufactured by Amano Pharmaceutical Co., Ltd.), Lipase AP-4 Amano (Derived from Aspergillus microorganisms; manufactured by Amano Pharmaceutical Co., Ltd.), Lipase MAP-10 (derived from Mucor microorganisms; manufactured by Amano Pharmaceutical Co., Ltd.), immobilized enzyme ribozyme (derived from Mucor microorganisms; manufactured by Novo Corporation), immobilized enzyme SP382 (Candida). Derived from a genus microorganism; manufactured by Novo Co.) and the like.

Examples of the enzyme-containing material include microbial cells, a culture solution or a treated product thereof, which has the ability to produce an enzyme having an activity of asymmetrically acylating compound (III). Examples of the microorganism having the ability to produce such an enzyme include Pseudomonas
Genus, Candida, Mucor
Genus, Aspergillus genus, Rhiz
opus), Bacillus, Arthrobacter
(Arthrobacter genus, Pichia genus, Chromobacterium genus, Humicola (Humicol)
a) Microorganisms belonging to genera such as Genus, Penicillium and the like can be mentioned.

Examples of the treated product of the culture solution include concentrates, dried products, additives of surfactants and / or organic solvents, and lysed enzyme-treated products of the culture solution. Examples of the treated bacterial cells include dried bacterial cells, surfactant and / or organic solvent additives, lytic enzyme-treated products, immobilized bacterial cells, or preparations of extracted enzymes from bacterial cells. As the organic solvent used, methanol, ethanol, propanol, alcohol solvents such as butanol, toluene,
It is possible to use aromatic solvents such as benzene, ether solvents such as diethyl ether and isopropyl ether, halogen solvents such as chloroform, solvents such as acetonitrile and dimethyl sulfoxide, or water saturated solutions or mixed solutions thereof. it can.

The compound (III) is, for example, when R ₁ is a benzyl group, Journal of the Medicinal Pharmaceutical Chemistry [(J. Med.
Pharm. Chem.), 1 , 73 (1959)]. The compound (III) as a substrate is used in an amount of 0.1 to 50% by weight, preferably 0.5 to 25% by weight, based on the reaction solution, and the compound (IV) is used in an amount of 0.025% based on the reaction solution.
~ 50% by weight, preferably 0.5 to 25% by weight,
Enzymes are used at 0.1 to 50% by weight, based on the weight of the substrate. The reaction is carried out in the range of 0 to 60 ° C, preferably 20 to 30 ° C. The reaction is usually completed in 1 hour to 1 day, though varying depending on the reaction temperature, the substrate concentration, the amount of enzyme and the like.

Isolation and purification of compound (I) and compound (II) from the reaction mixture can be carried out by filtration, concentration of enzyme, column chromatography and the like. The reaction can also be carried out by filling the column with an enzyme source and passing a solution in which compound (III) and compound (IV) are dissolved. The reaction solution obtained by the above-mentioned production method is not isolated or purified, but concentrated to sulfonylate the compound (II) contained in the residue,
Formula (V)

[0020]

[Chemical 9]

(Wherein R ₁ has the same meaning as defined above, X represents an alkylsulfonyloxy group or an arylsulfonyloxy group, and * represents that the carbon is optically active.) After conversion to
I)

[0022]

[Chemical 10]

By reacting with a metal salt of a carboxylic acid represented by the formula (wherein R ₂ has the same meaning as described above), a compound (I) having a reversed steric configuration can be obtained. By the treatment, the compound (I) having a desired configuration is obtained.
Can be obtained in high yield. The alkyl moiety of alkylsulfonyloxy has 1 to 7 carbon atoms.
And straight-chain alkyl such as methyl, ethyl, propyl, butyl, pentyl, hexyl and heptyl. Examples of the aryl moiety of arylsulfonyloxy include tolyl, phenyl, naphthyl and the like. Further, as the metal salt in the compound (VI), a lithium salt,
Examples thereof include sodium salt and calcium salt.

The conversion of the optically active 3-pyrrolidinol derivative obtained in the present invention into the optically active pyrrolidinol is carried out, for example, when R ₁ is a benzyl group, by contacting with palladium-carbon as a catalyst in methanol or acetic acid under hydrogen atmosphere. Achieved by giving back [Synth. Commun., 15 , 587 (198
Five) 〕.

The optically active 3-pyrrolidinol derivative thus obtained is (5R, 6S) -2 [(S)-
1- (acetimidoyl) pyrrolidin-3-ylthio]
-6-[(R) -1-hydroxyethyl] carbapene-
Β-lactam antibiotics such as 2-em-3-carboxylic acid and (3S) -1-benzyl-3-pyrrolidinylmethyl- (4S) -2,6-dimethyl-4- (m-nitrophenyl) ) -1,4-Dihydropyridine-3,5-dicarboxylate [Compound (VII); JP-A-60-19764) and other useful intermediates for the synthesis of dihydropyrrolidine compounds. For example, the compound (VII) is prepared from the optically active N-benzyl-3-pyrrolidinol obtained in Example 1 described later by Journal of Medicinal Chemistry [(J. Med. Chem.), 29 , 2504 (1986). ] It can manufacture by following the method as described in.

The present invention will be described below with reference to examples.

[0027]

【Example】

Example 1 Preparation of (3R) -N-benzyl-3-pyrrolidinol Racemic N-benzyl-3-pyrrolidinol, 500
Dissolve mg in 40 ml of isopropyl ether, vinyl acetate
0.6 ml (2.3 equivalents) and 160 mg of Lipase Amano P (Amano Pharmaceutical Co., Ltd.) were added. This reaction solution is kept at room temperature 25 ° C for 3
Stir for hours. The reaction solution was filtered and concentrated under reduced pressure, and the residue was purified by silica gel chromatography, eluting with a solvent in which ethyl acetate was added to n-hexane in an amount of 0 to 25% to obtain 300 mg of an oily compound. This compound had the following physical properties and was identified as N-benzyl-3-acetoxypyrrolidine. Yield 49%. ¹ H-NMR (CDCl ₃ ; 100MHz) δppm, 1.7-3.1 (6H, m),
2.03 (3H, s), 3.71 (1H, d, J = 12Hz), 3.76 (1H, d, J = 12H
z), 5.05-5.33 (1H, m), 7.22-7.48 (5H, m) IR (KBr) νmax (cm ^-1 ) 3110 (w), 3080 (m), 3050
(m), 2990 (s), 2940 (m), 2810 (s), 1745 (s), 1500
(m), 1485 (w), 1460 (m), 1450 (m), 1380 (s), 1260
(s), 1200 (m), 1160 (m), 1140 (m), 1090 (m), 1040
(m), 915 (m), 900 (w), 835 (w), 755 (s), 710 (s)

Further, the same column was eluted with a solvent obtained by adding 0 to 20% of n-hexane and 1% of triethylamine to ethyl acetate to obtain 184 mg of an oily substance. This compound had the following physical properties and was identified as N-benzyl-3-pyrrolidinol. ¹ H-NMR (CDCl ₃ ; 100MHz) δppm, 1.56-3.00 (6H, m),
3.64 (2H, s), 4.22-4.45 (1H, s), 7.08-7.62 (5H, m) IR (KBr) νmax (cm ^-1 ) 3375 (s), 3100 (s), 3080
(s), 3050 (s), 2970 (s), 2800 (s), 1500 (m), 1485
(m), 1460 (s), 1380 (m), 1355 (m), 1260 (m), 1215
(m), 1140 (m), 1100 (m), 1040 (m), 1050 (m), 920
(w), 895 (w), 750 (m), 705 (s)

N-benzyl-3-obtained in this way
The optical purity of acetoxypyrrolidine was measured by high performance liquid chromatography (HPLC) under the following conditions by hydrolyzing the acetoxy group with sodium hydroxide in methanol and using it as N-benzyl-3-pyrrolidinol. The optical purity of N-benzyl-3-pyrrolidinol was measured by HPLC as it was. As a result, N-benzyl-3-acetoxypyrrolidine was 100% ee within the detection limit of HPLC, unreacted N
-Benzyl-3-pyrrolidinol is 97% ee,
It was decided that they had mutually opposite configurations. HPLC measurement conditions Column: Daicel's chiral cell OD (4.6 mmφ × 2
Elution: n-hexane / 2-propanol / diethylamine = 95/5 / 0.1 Flow rate: 1 ml / min Detection: 254 nm UV absorption is measured

The absolute configurations of N-benzyl-3-acetoxypyrrolidine and recovered N-benzyl-3-pyrrolidinol were determined as follows. That is, N-benzyl-3-acetoxypyrrolidine obtained in the above reaction was hydrolyzed with sodium hydroxide in methanol to give N
-Benzyl-3-pyrrolidinol. N-benzyl-3-pyrrolidinol was added under hydrogen atmosphere in methanol,
De-N-benzyl was performed using 10% palladium-carbon as catalyst. The obtained 3-pyrrolidinol showed a specific optical rotation [α] _D = -32.2 ° (c = 3.55, methanol). Literature value: (R) body, [α] _D = −7.6 ° (c = 3.45,
Methanol) [Chemistry Letters (Chemistry L
etters), 893 (1986)]. Furthermore, the nitrogen of 3-pyrrolidinol was benzyloxycarbonylated to give N-benzyloxycarbonyl-3-pyrrolidinol. This compound exhibited a specific optical rotation [α] _D = -21.7 ° (c = 0.5, methanol). Reference value: (R) body, [α] _D = -18.2 ° (c = 0.5,
Methanol) [Acta Chemika Scandinavia (Act
a Chemica Scandinavia), 43 , 290 (1989)],
[Α] _D = -15.2 ° (c = 0.5, methanol) [Journal of Chemical Society Chemical Communications (J. Chem. Soc. Chem. Commun.), 1
298 (1980)]. From the above results, it was determined that the N-benzyl-3-acetoxypyrrolidine obtained by acetylation by the enzyme reaction had the absolute configuration (R). Therefore, the recovered N-benzyl-3-pyrrolidinol was determined to have the (S) configuration.

Example 2 Preparation of (3R) -N-benzyl-3-pyrrolidinol (2) Racemic N-benzyl-3-pyrrolidinol, 50 mg
Was dissolved in 5 ml of isopropyl ether and vinyl acetate 52 was added.
μl (2 equivalents) and 20 mg of lipoprotein lipase (Kyowa Medex) were added. This reaction solution is at room temperature 25 ° C.
It was stirred for 31 hours. The progress of the reaction was analyzed by HPLC, and acetylation proceeded at a conversion rate of 47% (3
S) -N-benzyl-3-acetoxypyrrolidine was produced.

Optical purity 35.2% ee. Unreacted (3R)-
The optical purity of N-benzyl-3-pyrrolidinol is 31.3.
% Ee (optical purity was measured in the same manner as in Example 1).

Example 3 Preparation of (3R) -N-benzyl-3-pyrrolidinol (3) Racemic N-benzyl-3-pyrrolidinol, 60 mg
Was dissolved in 2 ml of toluene, and 109 μl of vinyl acetate (3.5
Equivalent) and 20 mg of Lipase Amano P (manufactured by Amano Pharmaceutical Co., Ltd.) were added. The reaction solution was stirred at room temperature 25 ° C. for 6.5 hours.
The progress of the reaction was analyzed by HPLC, and acetylation proceeded at a conversion rate of 53% (3S) -N-benzyl-
3-acetoxypyrrolidine was produced. Optical purity 89%
ee. The optical purity of unreacted (3R) -N-benzyl-3-pyrrolidinol was 100% ee (optical purity was measured in the same manner as in Example 1).

Example 4 Production of (3R) -N-benzyl-3-pyrrolidinol (reaction by column) A mixture of 2.4 g of lipase amino P and 24 g of celite was suspended in a 9 mmφ × 200 mm column and charged in toluene. did. A racemic N-benzyl-3-pyrrolidinol was dissolved in toluene, and a mixed solution of a solution (substrate concentration: 50 mg / ml, vinyl acetate 3.5 equivalents) to which vinyl acetate was added was passed at room temperature of 25 ° C. The reaction liquid eluted from the column was concentrated and purified by silica gel column chromatography to obtain two types of oil substances of low polarity and high polarity. These substances have the same physical properties as in Example 1,
It was identified as (3R) -N-benzyl-3-acetoxypyrrolidine and (3S) -N-benzyl-3-pyrrolidinol. Tables 1 and 2 show the relationships among the flow rate, conversion rate, optical purity and column efficiency in this reaction.

[0035]

[Table 1]

[0036]

[Table 2]

Example 5 Preparation of (3S) -N-benzyl-3-pyrrolidinol with inversion of (R) -form in reaction solution 100 mg of racemic N-benzyl-3-pyrrolidinol
Was dissolved in 10 ml of t-butyl methyl ether, 0.2 ml of vinyl acetate was added, 35 mg of lipase amino P was added, and
The mixture was stirred at 5 ° C for 3 hours and 40 minutes. By filtering from the reaction solution,
The lipase was removed. 2 ml of tetrahydrofuran was added to the filtrate as it was, 90 μl of methanesulfonyl chloride and 170 μl of triethylamine were added, and the mixture was stirred at 20 ° C. for 20 hours. Saturated aqueous sodium hydrogen carbonate was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium hydrogen carbonate and saturated brine in that order, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. To the residue were added 2 ml of dimethyl sulfoxide, 250 mg of anhydrous sodium acetate, and the mixture was stirred at 120 ° C for 50 minutes. After cooling the reaction mixture, water was added, extraction was performed with ethyl acetate, the organic layer was washed 5 times with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Dissolve the residue in ether, 0.5 mm
On a thin layer chromatography (TLC) plate of
Hexane / ethyl acetate = 1/1 was developed, the target substance was scraped off, extracted with ethyl acetate, and concentrated under reduced pressure. The residue was dissolved in 2 ml of methanol, 0.5 ml of 2N aqueous sodium hydroxide solution was added, and the mixture was stirred at 25 ° C. for 1 hour. Water was added to the reaction solution, and extraction was performed with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 83 mg of an oily substance. Yield 83%. This product had the same physical properties as N-benzyl-3-pyrrolidinol of Example 1 and was identified as N-benzyl-3-pyrrolidinol. Also,
As a result of measuring the optical purity by the same method as in Example 1,
It was decided to be 91% ee.

[0038]

INDUSTRIAL APPLICABILITY According to the present invention, an optically active 3-pyrrolidinol derivative, which is an important intermediate of an optically active compound useful as a drug such as β-lactam antibiotics and dihydropyridine compounds, can be produced in high yield. You can

Claims (1)

[Claims] 1. Formula (III): (In the formula, R ₁ represents a substituted or unsubstituted benzyl group, alkanoyl group or aroyl group.) [Hereinafter abbreviated as compound (III). ] And the formula (IV) (R ₂ represents an alkyl group or an aryl group, and R ₃
Represents hydrogen, an alkyl group, an alkenyl group, an alkanoyl group or an aroyl group. ) And a compound represented by
Compound (III) is reacted in the presence of an enzyme source having an activity of stereoselectively acylating to give a compound of formula (I) (Wherein R ₁ and R ₂ have the same meanings as described above, and * means that the carbon is optically active.) And the formula (II) (In the formula, R ₁ has the same meaning as described above, and * means that the carbon is optically active.) 3
-A method for producing an optically active substance of a 3-pyrrolidinol derivative, which comprises obtaining an optically active substance of a pyrrolidinol derivative.