JPH0240343A - Fluorine-containing polyester and production thereof - Google Patents

Abstract

NEW MATERIAL:The fluorine-containing polyester of formula I, II, III or IV (Rf<1> and Rf<2> are same or different alkyl containing fluorine atom; n is integer of >=2). EXAMPLE:Ethyl-3-trifluoromethyl-3-hydroxypropanate polymer. USE:It has physiological activity originated from the presence of fluorine- containing alkyl group and optical activity and is useful as raw materials for various physiologically active substances (e.g., vitamin-analog compound, cardiotonics, antibiotic substance or antibacterial agent) and polymeric compounds. PREPARATION:The compound of formula I - formula IV can be produced by reacting a fluorine-containing compound of formula V or formula VI with enzyme in an organic phase. The enzyme is a lipase produced by microorganisms of genus Candida, genus Pseudomonas, etc., especially a microbial lipase composed of a macromolecule. The reaction temperature is preferably 50-0 deg.C.

Description

Detailed Description of the Invention B. Industrial Application Field The present invention relates to a fluorine-containing polyester and a method for producing the same, and particularly relates to an optically active fluorine-containing polyester (including cyclic polyesters or lactones) and a method for producing the same. .

Previous Art Hydrolytic enzymes as chiral catalysts for asymmetric hydrolysis have been studied for many years [Enzymic and Non-Enzymic Catalysis] by P. Dunnil et al. 1980), ``Stereospecificity in Organic Chemistry and Enzymology'' by J. L. Lee et al.
82) ). However, the study of the ability of the above enzymes for the asymmetric synthesis of fluorinated compounds remains untouched from a practical point of view. In addition, synthetic methods for obtaining various versatile chiral fluorinated compounds with high optical purity have not been investigated in detail.

In particular, it is extremely difficult to obtain an optically active polyfunctional fluorine-containing compound with high optical purity, and the reality is that it has hardly been studied.

C.1 Objective of the Invention An object of the present invention is to provide an optically active polyester in which fluorine atoms are introduced at desired positions.

Another object of the present invention is to produce optically active fluorine-containing polyesters with high purity and high yield in a relatively short period of time under mild conditions through an asymmetric organic synthesis reaction that utilizes the latent functions of enzymes. The objective is to provide a method that can be used to manufacture

22 Structure of the invention and its effects, that is, the present invention is based on the following formulas [I], (II), [■]
or (IV).

General formula [I]: Rf' -(-0-CH-CH2CCL)-T −Mathical formula [■]d -+OCR.

General formula [■]: Rf" CH-CO+-T - Formula [■]: (However, the above - formula [1), (II), (1) or (I
In V), Rf', Rf2: the same or different alkyl groups containing a fluorine atom, n: an integer of 2 or more. ) The present invention also provides a method for producing the above-mentioned fluorine-containing polyester, in which the following - Formula (N, (II), (III) or (IV)
The present invention also provides a method for producing a fluorine-containing polyester that obtains a fluorine-containing polyester represented by the following formula.

General formula [l]: Rf' -(-0-CH-CHffi C0 square-formula [■]
: Rf" 1 + OCH, -CH-C0b General formula []]: Or - mathematical formula [■]: ., jl Uhz General formula [■]: (However, the above - mathematical formula (1), (II), (III) , [
(2)], [V) and (Vl), R', R2: hydrogen atom, or an alkyl group with or without a substituent (R' and R2 may be the same or different from each other), Rfl, Rf": the same or different alkyl groups containing a fluorine atom, n: an integer of 2 or more.) In the present invention, the above "in the organic phase" refers to the formula [V]
, - In addition to cases in which the compound (substrate) represented by the formula (Vl) is dissolved in various organic solvents, or reacted in a suspended or dispersed state, the compound (substrate) represented by the general formula (Vl) can be reacted without using an organic solvent.
], -The meaning also includes cases where one or both of the compounds represented by the formula (Vl) are used as a medium (as a solvent or as a dispersion medium). In the search for biologically active substances, stereochemical discrimination between substrates and living organisms is important, and from this stereochemical perspective, the present inventors utilized biochemical techniques to identify the inactivation of useful organic compounds. I have been working on the problem of simultaneous synthesis.

In the course of such research, the present inventors realized asymmetric synthesis of fluorine-containing polyester compounds by utilizing the new functional expression of hydrolases. That is, according to the present invention, an optically active fluorine-containing polyester compound can be obtained with extremely high purity and high yield under mild conditions.

Conventionally, enzymes have generally been characterized by expressing their functions using water as a medium, but from the standpoint of organic synthesis of useful compounds, it is advantageous to express their functions in an organic solvent. As a result of intensive studies, the present inventors have found that even if the asymmetric aldol type reaction is carried out in the presence of an enzyme in an organic phase such as an organic solvent, the carbon-carbon bond forming reaction proceeds sufficiently and easily. This is what led us to the present invention.

In particular, this reaction is extremely useful in that it is the first time that an enzyme has been used to synthesize an optically active fluorine-containing polyester compound in an organic phase.

General formulas (1) and [11] synthesized by the method according to the present invention
, (III), and (TV) have a fluorine-containing alkyl group and have optical activity, so they exhibit physiological activity and are useful for various biologically active substances (vitamin analogs, cardiotonic agents, antibiotics, etc.). antibacterial agents, etc.) and raw materials for polymer compounds.

As Rf' and Rf7 in the above formula, the number of carbon atoms is 10 or less (for example, CFy, Cz Fs, C3F'
r, C4F9, etc.).

In addition, n is 2 or more, but especially polyester such as oligomer (molecules 17,000 to 20,000, e.g. 10,000 to 10,000), 5
The number should be equivalent to 1,000,000,000.

In the method according to the present invention, Rf' and Rf''n are the same as described above, and the general formula [■], (V
In [), at least one of (7) R1 and RZ (7) is preferably an alkyl group having 10 or more carbon atoms, more preferably 5 or less, in view of molecular size, etc. Furthermore, any substituent can be selected as long as it does not inhibit the polyester synthesis reaction.

In the method according to the invention, enzymes such as Can
dida genus, Pseudon+onas genus, ^lca
Lipases produced by the genus Igenes, the genus Achromobacter, etc. can be used. Among these, microbial lipases consisting of macromolecules are particularly preferred for producing the fluorine-containing polyester compound in the organic phase according to the present invention.

That is, it has been discovered that when such macromolecular microbial ribasese is used, the reactions of reaction formulas (1) and (2) described below can occur satisfactorily in an organic reaction system, whereas such reactions hardly occur in an aqueous system.

Specific examples of such macromolecular microbial lipases include:
For example, Alcaligenes (a name belonging to the genus Mμ)
Ji P L-266 (Alca order ness, P L
-266: Lipase (refer to Japanese Patent Publication No. 58-36953) (hereinafter referred to as lipase PL-266) produced by Kaikouken Bokuyori No. 3187), and scaly PL-679, which also belongs to alkaline earth metals. Entan Danguchi rib row”mu PL-
679: Microtechnical Research Institute No. 3783) (see Japanese Patent Publication No. 60-15312) (hereinafter referred to as lipase PL-679), and Achromobacter (A
chromobacter) genus AL-86
5. Lipase produced by Yayuyu AL-865: @ Koken Meiyori No. 1213)
080) (hereinafter referred to as lipase AL).

Table 1 compares the molecular weight and optimum pH of macromolecular microbial lipases that can be used in the present invention.

As shown in Table 1, alkaline microbial lipases consisting of macromolecules with a molecular weight of 100,000 or more are particularly suitable as lipases that can be used in the present invention.

It is not clear why such macromolecular lipases, etc. cause the above reaction in the organic phase, but it is likely that macromolecular lipases, etc. not only have their active groups protected by a number of subunits, but also protect their molecules. It is thought that this is not due to the large amount of intramolecularly bound water necessary for the reaction to occur.

It can be inferred that this has some relation to the fact that the enzymatic reactions of reaction formulas (1) and (2) of No. 41 are possible in an organic solvent.

Therefore, there are no restrictions on the microbial origin or type of enzymes other than those exemplified as long as they cause the above reaction.

Further, the enzyme used in the present invention may be a purified product or a crude product, and its usage form may be an enzyme powder alone or a granular enzyme shaped with a suitable binder.

In addition, if necessary, it may be hardened by adding a suitable diluent other than the enzyme as a diluent.Furthermore, by immobilizing the enzyme on various carriers, the efficiency of the reaction and the usability of the enzyme will be further increased. If so, it can also be used by being supported and immobilized. Examples of such immobilization carriers include polypropylene membranes, DEA E-Toyo pearls,
Sepabeads FPDA (DEAE manufactured by Mitsubishi Kasei Corporation), CM
-Cellulose, DEAE-cellulose, various polymers such as ion exchange resins and adsorption resins such as Amberlite IRA68, IRA938, IRA93, and IRA94, bentonite, etc. can be used, and after being supported and immobilized on these, it is dried. It can be used. These drying methods can be carried out by methods such as freeze drying, immersion in a drying solvent such as acetone, and washing.

The activity of microbial lipase used in the present invention was measured by the following method. Lipase PL679 and PL-
For 266, Kunio et al.'s method (Agric.

Biol, Chen+, 46(5), 1159, 19
82), and for lipase AL, the method of Kunio et al. (Oil Kagaku 23(2), 98.1974) was used. The lipase activity determined by the above method was Lipase P L-679 powder (manufactured by Polysaccharide Sangyo Co., Ltd.) = 100,000 single blade g, Lipase P L-
266 powder (manufactured by Polysaccharide Sangyo Co., Ltd.): 1.1 blades/g; Lipase AI powder (manufactured by Polysaccharide Sangyo Co., Ltd.): 15,000 ri/g.

There are no particular restrictions on the amount of lipase used, but
An example of a usage amount is about 10 to 100,000 units per g. It is also possible to use two or more types of reparsers together.

It is preferable to set the reaction temperature at 50°C or lower in order to effectively exert the action of the enzyme, and it is also preferable to set the reaction temperature at 0″C0 or higher.To promote the reaction, it is preferable to set the reaction temperature at 25°C or higher. The pressure during the reaction may be several atmospheres or less, particularly normal pressure, and the reaction time may be within several days.

Organic solvents used in the reaction include benzene, hesaquin,
Pentane, trichlorotrifluoroethane, octane,
Ethanol, propatool, butanol, pentanol, hexanol, heptatool, octatool, nonanol, methanol, propatool, 2-propatool,
Suitable solvents include nonpolar solvents such as 2-butanol, 2-pentanol, decanol, dodecanol, tetrahexanol, hexadecanol, and octadecanol.

Next, the fluorine-containing polyester according to the present invention will be explained in more detail along with its synthesis method.

That is, the present inventor carried out a highly stereoselective polyester synthesis reaction using a polymeric M (molecular weight: 300,000 to 400,000) lipase, such as the above-mentioned lipase active system or lipase AL system, as an enzyme, and 3 having a fluorine alkyl group (e.g. trifluoromethyl group (CF) at the α or β position
-ethyl hydroxypropanoate 1 (3-)rifluoromethyl-3-hydroxypropanoate) or 2 (
2-) Ethyl fluoromethyl-3-hydroxypropanoate) was used to react with the enzyme in various organic solvents.

It has been found that the optically active (chiral) polyester obtained in this reaction varies depending on the lipase used, as described below.

υ Yuυ Rule According to this reaction, when a lipase production source is applied to the Tatoki, the molecular weight is 10,000 to 1,000, using a solvent system such as benzene or hexane.
An optically active linear polyester main or i (oligomer) having a position of 50,000 was obtained. Unreacted monomers (hydroxy esters) were recovered from the system, and these 11 monomers were, for example, (S) monomers or (S) rods with optical purity >70%ee (conversion rate of 41%). When the conversion rate is >75%ee, when the conversion rate is 62%, it is >95%ee).

On the other hand, when the lipase AL system is applied, trilactone or main trilactone, which is a polyester with a cyclic structure, and (
Monomer y of the R) form or (R) suspension (optical purity>92%ee when the conversion rate was 63%) were obtained, respectively.

According to the above-mentioned reaction, the synthesis reaction proceeds stereoselectively due to the action of the lipase used, and a chiral polyester having a fluorine atom introduced at the desired position can be obtained with high efficiency. The mechanism by which this regioselectivity occurs is not clear, but it is thought to be related to the shape of the enzyme's active site and the interaction between the enzyme's active site and fluorine-containing alkyl groups and carbonyl groups. .

The present inventor investigated the influence of various lipases on the above reaction formula (2) (oligomerization and/or lactonization). The results are shown in Table-2.

Table-2 Lipase production source Candida c.
us 'a onicus ”Rh1zo u
s delo+er ``81 cali enes
s, "Yield 1) (%) In the above table, a) 1 g of lipase and ethyl-2-trifluoromethyl-
3-Hydroxypropanate was reacted in dry benzene 30 at 40-41°C.

b) Manufactured by Meito Sangyo Co., Ltd., produced by Lipase MY.

C) Manufactured by Nagase Biochemical Co., Ltd.

d) Manufactured by Sigma.

e) Manufactured by Meito Sangyo Co., Ltd.

f) Lipase PL-679 produced by Meito Sangyo Co., Ltd.

From the results in Table 2, lipase M Y , and Pseu
While lipase F obtained from S. domonas acts as a catalyst for the lactonization reaction, some other lipases do not exhibit such activity. However, such lactonization (condensation reaction) is not common to lipases, and in particular, lipase PL-679 and lipase AL, which are suitable for the present invention, are selected to mainly cause linear oligomerization (former) and lactonization (latter). It turns out that there is a gender.

Next, the present inventor conducted the above reaction using lipase PL-679, and at that time investigated the influence of the solvent used. The results are shown in Table-3. However, in the reaction, lipase PL
-6793 g and 20 mmol of substrate in 50 d of organic solvent.
The mixture was stirred for 72 hours at 40-41°C. The resulting product was analyzed by gel permeation chromatography (GPC) in tetrahydrofuran at 50°C. This analysis was performed using a refractive index measuring device (Shodex R1).
Shodex with GP CK
It was performed using Shimadzu LC-5A high performance liquid chromatography using an F-803 column (flow rate was 1.51ffi/wi
n).

(Left below) From the second result, nonpolar solvents such as benzene, hexane, and 1,1°2-trichlorotrifluoroethylene are effective for the condensation reaction. Furthermore, the molecular weight distribution of the obtained polyester is within a narrow range, and its degree of polymerization can be controlled by selecting the type of enzyme and reaction medium.

E. Examples Hereinafter, the present invention will be explained in more detail with reference to Examples.
The following examples illustrate the present invention, and various changes can be made based on the technical idea of the present invention.

(Polymerization of ethyl-3-trifluoromethyl-3-hydroxypropanate) 3 g of lipase P L-679 (...■Danfu Lori Chu 1, manufactured by Meito Sangyo Co., Ltd.) and ethyl-3-trifluoromethyl-3-hydroxypropanate. Fluoromethyl-3-hydroxypropanate 3.7 g (20 mmo
l) and were stirred in 50 ml of benzene at 40-41°C for 72 hours. The presence of residual ethyl-3-trifluoromethyl-3-hydroxypropanate was detected by "FNM"
R signal intensity (C6Hs CF3 as internal standard (62%
). ), and then the polyester and hydroxyester were separated by column chromatography using benzene as an eluent. Polyester is 50°
Gel permeation in tetrahydrofuran at C.
Subjected to chromatography (cpc). The analysis was carried out using a Shodex (Shodex R1) equipped with a refractometer (Shodex R1).
Shimadzu LC using GPCKF-803 column
-5A high performance liquid chromatography. The flow rate is 1
．． It was 5mff1/min. The absolute configuration of the remaining (-)-ethyl-3-trifluoromethyl-3-hydroxypropanate was the (S)-enantiomer, and its optical purity was >95% ee.

The analytical data for each of the above products were as follows.

Polyester: "F NMR (benzene: C6H6): δ-11,7 (
d, ffcF: +-Ctl=8.0Hz) (external standard:
ChCO□11) 'HNMR (di C4H6): 62.40 -2.
55 (2H, m), 4.30 -4.45 (IH, m) IR (at+-'): 1725 (C=0) hydroxy ester: bp: 70 "C/15 Hg 19F NMR (CDCl2): δ 2.6 (d,, 7CFx-CI=6.6Hz) (external standard: CF3C0ZH) 'HNMR (CDCI+): δ 1.25 (CI (
3,t, JC1+3-CIIZ=7.3Hz), 2.
62 (CIIZ, dJcH2-CI = 5.6Hz
), 4.30 (4H, m) IR (cm-'): 3580
(OH), 1720 (C=O)(α) o:
-20,5 (neat) Actual drawing Itsuiri (polymerization of ethyl-2-trifluoromethyl-3-hydroxypropanate) Lipase PL L-679 (Tanbao Dansara Tanjihe, manufactured by TG Sangyo Co., Ltd.) ) and 3.7 g (20o+
mol) in benzene 50d at 40-41°
The mixture was stirred at C for 72 hours. The presence of residual ethyl-2-trifluoromethyl-3-hydroxypropanate was
NMR signal intensity (C6HSCF) was internally referenced fjA (
64%). ), and then the polyester and hydroxyester were separated by column chromatography using benzene as an eluent. The polyester was subjected to gel permeation chromatography (GPC) in tetrahydrofuran at 50°C. The analysis is
It was performed on a LC-5A high performance liquid chromatography using a Shodex GPCKF-803 column with a refractometer (Shodenox R1). The flow rate was 1.5 ml/min. The absolute configuration of the remaining (=)-ethyl-2-trifluoromethyl-3-hydroxypropanate was the (S)-enantiomer, and its optical purity was >95% ee.

The analytical data for each of the above products were as follows.

Polyester: 19F IJMR (Benzene: C, I+,): δ-
9,7 (d, JCFs-CII=8.0IIz) (external standard: ChCOzll) 'II NMR (d, -C6116): δ 3.20
-3.70 (ill, m), 4.43 4.50 (2
H, +m) IR (cr'): 1720 (C=0) hydroxy ester; bp: 59-60°c/ 3mm 11g"F NMR (
CC14): δ-12,0(d, JCFz-CI
I=8.611z) (external standard 2CF3CO, 11)' II NMR (CCI 4): δ1.3HCI!
3. L,, JClls-CIIz=7.8112), 2
．． 15 (Ill).

3, 24 (Cll, d, d, q JCI
I-11a=4.911z, JTC! (-11b=7
．． 6112) 3.84 (C11allb, d, d, JH
a-Hb=11.311z), 4.04(C11a11
b, d, d), 4.19 (Cll2. (1) IR (cm
=): 1”140 (C=0), 3470 (Oll) (c
r), /Me011: -3,90(GO,9
2) Next Grade 1 Polyester and hydroxyester were obtained in the same manner as in Example 1 except that the substrate monomer was changed as shown in Table 4 below.

LIPase A L (Polymerization of ethyl-3-trifluoromethyl-3-hydroxypropanate) L (Achromobacter s
,, manufactured by Meito Sangyo Co., Ltd.) 3 g and 3.7 g of ethyl-3-trifluoromethyl-3-hydroxypropanate.
(20 mmol) in 50 d of benzene from 40 to
Stirred at 41°C for 72 hours.

The presence of remaining ethyl-3-trifluoromethyl-3-hydroxypropanate was determined by I9F NMR signal intensity (
C6H, CF is used as an internal standard (63%). ), and then the cyclic polyester and hydroxyester were separated by column chromatography using benzene as an eluent. The polyester was subjected to gel permeation chromatography (GPC) in tetrahydrofuran at 50°C. The analysis was performed using a Shodex G with a refractometer (Shodex R1).
It was performed on a High Performance Liquid Chromatography LC-5A using a PCK F-803 column. Flow rate is 1.5 d
/sin. The absolute configuration of the remaining (+)-ethyl-3-trifluoro'methyl-3-hydroxypropanate is the (R)-enantiomer, and its optical purity is 〉. It was 92%ee.

The analytical data for each of the above products were as follows.

Polylactone: "F NMR (CDC13): δ -7,
9 (d, JCFs-C11=7.511zn (external standard: CF3C0zll) It NMR (CDCIs): δ2.35-2
．． 42 (211, n+), 4.20-4.35 (III
, m) IR (cm-'): 1725 ((:=O) hydroxy ester; bp: 70-71'C/ 5mm11g'9F NM
R(CDCh): δ2.6(dJcFa-C1l=6
．． 611z) (external standard: ChCO□11) If NMR (CDC13): δ1.25 (CIl
i, t, J (:1h-CIli =7.311z),
2.62 (C11t, dJCllz- to l1=5.61
1z), 4.30(411,n+)Ill(cm-li:
3580 (O1+), 1720 (C=0) [α) n
: +20. Hneat) Lipase AL (Polymerization of ethyl-2-trifluoromethyl-3-hydroxypropanate) Lipase AL (^cnromobacter s,,
3 g of Meito Sangyo Co., Ltd.) and 3.7 g of ethyl-2-trifluoromethyl-3-hydroxypropanate (manufactured by Meito Sangyo Co., Ltd.).
20 mmoffi) and 40 to 4 in benzene 50
Stirred at 1°C for 72 hours.

The presence of residual ethyl-2-trifluoromethyl-3-hydroxypropanate was determined by '9F NMR signal intensity (
After confirmation using C, HSCF3 as an internal standard (61%), the cyclic polyester and hydroxy ester were separated by column chromatography using benzene as an eluent. Polyesters were subjected to gel permeation chromatography (CPC) in tetrahydrofuran at 50"C. Analysis was performed using a Shodex with a refractometer (Shodex R1).
Quality LC-5 using G PCK F-803 column
A: Performed by high performance liquid chromatography. Flow rate is 1.5
ift/min. Remaining (+)-ethyl-
The absolute configuration of 2-trifluoromethyl-3-hydroxypropanate was the (R)-enantiomer and its optical weight was >94% ee.

The analytical data for each of the above products were as follows.

Polylactone: 19F NMR (CDCIz): δ-8,5 (d,,
7CFx-CIl=8.511z) (external standard: C1
', C0ZII) It NMR (CDC13): δ 3.25 3
．． 65 (ill, m), 4.40 -4.55 (211
, m) IR (cm -' ): 1720 (C=0) hydroxy ester: bp: 59-60'C/ 3m+n11g1'F NF
IR(CCL): δ-12,0(d,7CFff-
CII=8.611z) (external standard: CFzCOzl
l) 'II NMR (CC1,): δ 1.31 (C1h,
t, JcII+-C1lz =7.811z), 2.
15 (+++), 3, 24 (C1l, d, d
, q JCII-11a=4.911z ,,7C
1l-11b=7.611z), 3.84(C11a1
1b, d, d, Jlla-11b=11.311z),
4.04 (C1llalb, d, d), 4.19 (C
1lz, Q) IR (cm-'): 1740 (C=O)
,3470(011)Ccr]n/MeOll :+
3.88 (C1,02) λ Group 11 A polylactone and a hydroxy ester were obtained in the same manner as in Example 4, except that the substrate monomer was changed as shown in Table 5 below.

(Margin below)

Claims (1)

[Claims] 1. A fluorine-containing polyester represented by the following general formula [I], [II], [III] or [IV]. General formula [I]: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ General formula [II]: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ General formula [III]: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ General formula [IV]: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ {However, the above general formula [I], [II], [III] or [I
V], Rf^1, Rf^2: the same or different alkyl groups containing a fluorine atom, n: an integer of 2 or more. } 2. General formula [V]: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ Or general formula [VI]: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ Adding enzymes to the fluorine-containing compound represented by By acting, the following general formulas [I], [II], [III]
Or a method for producing a fluorine-containing polyester to obtain a fluorine-containing polyester represented by [IV]. General formula [I]: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ General formula [II]: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ General formula [III]: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ General formula [IV]: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ {However, the above general formulas [I], [II], [III], [IV]
, [V] and [VI], R^1, R^2: hydrogen atom, or an alkyl group with or without a substituent (R^1 and R^2 may be the same or different) ), Rf^1, Rf^2: the same or different alkyl groups containing a fluorine atom, n: an integer of 2 or more. }