EP1409704A2 - Verfahren zur enantioselektiven reduktion von prochirale ketoverbindungen die mindestens ein trifluormethylverbindung auf dem aromatischen zyklus haben - Google Patents

Verfahren zur enantioselektiven reduktion von prochirale ketoverbindungen die mindestens ein trifluormethylverbindung auf dem aromatischen zyklus haben

Info

Publication number
EP1409704A2
EP1409704A2 EP02758523A EP02758523A EP1409704A2 EP 1409704 A2 EP1409704 A2 EP 1409704A2 EP 02758523 A EP02758523 A EP 02758523A EP 02758523 A EP02758523 A EP 02758523A EP 1409704 A2 EP1409704 A2 EP 1409704A2
Authority
EP
European Patent Office
Prior art keywords
process according
ketone compound
enzyme
lactobacillus
reaction
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP02758523A
Other languages
English (en)
French (fr)
Inventor
Claude Bensoussan
Mirjana Gelo-Pujic
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Rhodia Chimie SAS
Original Assignee
Rhodia Chimie SAS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Rhodia Chimie SAS filed Critical Rhodia Chimie SAS
Publication of EP1409704A2 publication Critical patent/EP1409704A2/de
Withdrawn legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/22—Preparation of oxygen-containing organic compounds containing a hydroxy group aromatic

Definitions

  • the subject of the present invention is a method of enantioselective reduction of a prochiral aromatic ketone comprising at least one trifluoromethyl group on the aromatic ring, according to a biocatalysis or bioconversion method.
  • optically active phenylalkanolic compounds and in particular (R) or (S -1-phenylethanol, are compounds widely used as synthons in the field of pharmacy and agrochemistry.
  • the objective is to obtain the enantiomer having the desired property and to minimize the formation of the other enantiomer.
  • the described method leading to an alcohol (S), the objective of the invention is to provide a desired optically active alcohol of configuration (R) according to a method of enantioselective reduction of the corresponding ketone.
  • a method of enantioselective reduction of a prochiral aromatic ketone comprising at least one trifluoromethyl group on the aromatic cycle characterized in that the reduction is carried out in the presence of the enzyme from Lactobacillus.
  • the preferred enzyme used according to the invention is the enzyme originating from Lactobacillus Kefiri.
  • the method of the invention provides predominantly access to the alcohol of configuration (R).
  • ketone compound denotes the starting substrate, namely, the prochiral aromatic ketone comprising at least one trifluoromethyl group on the aromatic cycle. According to the invention, the reduction is carried out in the presence of an enzyme of the alcohol dehydrogenase type.
  • a first variant of the invention consists in using the isolated enzyme. Another variant of the invention is to bring into play a biomass comprising said enzyme.
  • ketone compound corresponding to the general formula is used in the process of the invention:
  • Ri represents an alkyl, alkenyl, cycloalkyl, aryl or arylalkyl group
  • - n is a number at least equal to 1, preferably between 1 and 3, - at least one trifluoromethyl group is in position 3, 4 or 5.
  • alkyl means a linear or branched hydrocarbon chain having from 1 to 15 carbon atoms and preferably from 1 or 2 to 10 carbon atoms.
  • alkenyl is meant a hydrocarbon group, linear or branched having from 2 to 15 carbon atoms, comprising one or more double bonds, preferably 1 to 2 double bonds.
  • cycloalkyl is meant a cyclic hydrocarbon group, comprising from 3 to 8 carbon atoms, preferably a cyclopentyl or cyclohexyl group.
  • aryl is meant an aromatic mono- or polycyclic group, preferably mono- or bicyclic comprising from 6 to 12 carbon atoms, preferably phenyl or naphthyl.
  • arylalkyl is meant a hydrocarbon group, linear or branched carrying a monocyclic aromatic ring and comprising from 7 to 12 carbon atoms, preferably benzyl.
  • Ri represents an alkyl group having from 1 to 4 carbon atoms, preferably 1 or 2, - and n is a number equal to 1 or 2.
  • the reduction is carried out by using preferentially, the enzyme originating from Lactobacillus Kefiri.
  • the microorganism is a collection microorganism DSM20587.
  • the reduction can be carried out in the presence of the isolated enzyme or of a biomass containing it.
  • the enzyme is introduced into a buffered medium having a pH of around 7, preferably obtained with a phosphate buffer comprising 0.1 mol / l of mono- and dipotassium phosphate.
  • a co-factor is added, namely, NADPH (nicotine-adenine-dinucleotide-phosphate).
  • NADPH nicotine-adenine-dinucleotide-phosphate
  • the final concentration of the co-factor in the final medium is preferably between 0.1 to 1 mmol / l.
  • the co-factor which undergoes oxidation during the reaction is regenerated by conventional means such as, for example, the use of a secondary alcohol, preferably cyclopentanol or l isopropanol.
  • the amount of alcohol used is in excess of the stoichiometry of the substrate. It is determined so that the concentration of alcohol in the medium is between 20 to 100 mmol / l.
  • the ketone compound to be reduced is then introduced. It is implemented at a concentration advantageously between 5 to 20 mmol / l.
  • the reaction is carried out at a temperature preferably between 30 ° C and 37 ° C.
  • the reaction is carried out at atmospheric pressure and the reaction medium is preferably stirred.
  • the medium is kept under stirring for a period which can be very variable. It is most often between 6 and 24 hours.
  • the optically active alcohol obtained is separated in the usual way, for example, by carrying out an extraction using a suitable organic solvent such as, for example, dichloromethane, ethyl ether, ethyl acetate or any other suitable solvent.
  • a suitable organic solvent such as, for example, dichloromethane, ethyl ether, ethyl acetate or any other suitable solvent.
  • the enzyme contained in the cells of the microorganism is used.
  • a first step in the process of the invention consists in carrying out the fermentation of the strain Lactobacillus Kefiri, in a conventional medium used to cultivate Lactobacilli.
  • a second step is to carry out the reduction reaction in the presence of the biomass previously obtained.
  • a fermentation medium comprising, for example, a carbon source, a nitrogen source and mineral salts.
  • carbon sources mention may in particular be made of maltose, glucose, sucrose, lactose, glycerol, starch, sorbitol, mannitol, propylene glycol.
  • yeast extracts beef extracts, peptone, ammonium sulphate, ammonium citrate, sodium nitrate or any other source of nitrogen containing amino acids.
  • mineral salts can be added and in addition to those mentioned as a nitrogen source, there may be mentioned sodium acetate, magnesium sulfate, manganese sulfate or potassium phosphate. Reference will be made to the examples to illustrate the composition and the concentrations of the various constituents of the fermentation medium.
  • Lactobacilli The cultivation of Lactobacilli is carried out anaerobically or pseudo-anaerobically and the skilled person knows how to conduct fermentation under these conditions. From a practical point of view, the fermentation medium is seeded with an inoculum of Lactobacillus, preferably Lactobacillus Kefiri which is present, most often in the form of an aqueous suspension which may contain a cryo-protective agent for example , dimethylsulfoxide or glycerol at a concentration, for example, from 10 to 20% by weight. Fermentation takes place at a pH advantageously between 6 and 7 and at a temperature preferably between 25 to 30 ° C.
  • a cryo-protective agent for example , dimethylsulfoxide or glycerol
  • the fermentation time is generally 2 to 4 days and is most often 3 days.
  • the biomass is recovered in a conventional manner. For example, a centrifugation can be carried out for approximately 3 to 15 min, then the supernatant is removed by decantation and a pellet is recovered, which is most often washed with physiological water.
  • the Lactobacillus biomass obtained is 1 to 5 g / l expressed in dry cells.
  • the enantioselective reduction of the ketone compound is then carried out in the presence of the Lactobacillus biomass expressing the activity of alcohol dehydrogenase.
  • the biomass is introduced in an amount of 10 to 30 g of dry matter per liter of reaction medium also comprising a buffer.
  • a buffered medium having a pH of approximately 7 is chosen as above, preferably obtained with a phosphate buffer comprising 0.1 mol / l of mono- and dipotassium phosphate.
  • the ketone compound to be reduced is then introduced. It is implemented at a concentration advantageously between 5 to 20 mmol / l.
  • the reaction is carried out at a temperature advantageously between 30 ° C. and 37 ° C.
  • the reaction is carried out at atmospheric pressure and the reaction medium is preferably stirred.
  • the medium is kept under stirring for a period which can be very variable. It is most often between 6 and 24 hours.
  • the biomass is separated in a conventional manner, for example by centrifugation.
  • the supernatant comprising the optically active alcohol is recovered and separated, in a usual manner for example, by carrying out an extraction using a suitable organic solvent such as, for example, dichloromethane, ethyl ether, ethyl acetate or any other suitable solvent.
  • a suitable organic solvent such as, for example, dichloromethane, ethyl ether, ethyl acetate or any other suitable solvent.
  • the optically active alcohol can be obtained from a prochiral ketone with an excellent yield and a very high enantiomeric excess of up to 100%.
  • the alcohol obtained mainly is of configuration (R).
  • the strain Lactobacillus kefiri DSM20587 is cultivated in quasi-anaerobic static culture in the Man-Rogosa-Sharp medium (MRS) described below at 25 ° C, for 72 hours.
  • MRS Man-Rogosa-Sharp medium
  • the culture medium [MRS medium (Difco-0881)] has the following composition: - Peptone n ° 3 10 g
  • the pH is 6.5.
  • the medium is first of all sterilized in a conventional manner by heating in an autoclave at a temperature of 121 ° C., for 20 min.
  • the medium (100 ml) is seeded with 0.1 ml of a cell suspension of Lactobacillus Kefiri socked in glycerol water (15% glycerol) and incubated for 72 hours at 25 ° C.
  • the biomass obtained is 1.5 g / l expressed in dry cells.
  • the cells obtained are washed with physiological water.
  • BTA 3,5-bis (trifluoromethyl) acetophenone
  • the enantiomeric excess ee expressed in% which is the ratio between [(R) - (S)] and [(R) + (S)] x 100, is determined by gas chromatography (GPC) on the chiral column Chiralsil Dex CB: 100 ° C to 160 ° C with a speed of 2 ° C per min.

Landscapes

  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
EP02758523A 2001-07-02 2002-06-28 Verfahren zur enantioselektiven reduktion von prochirale ketoverbindungen die mindestens ein trifluormethylverbindung auf dem aromatischen zyklus haben Withdrawn EP1409704A2 (de)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
FR0108742 2001-07-02
FR0108742A FR2826650A1 (fr) 2001-07-02 2001-07-02 Procede de reduction enantioselectif d'une cetone aromatique prochirale comprenant au moins un groupe trifluoromethyle sur le cycle aromatique
PCT/FR2002/002251 WO2003004666A2 (fr) 2001-07-02 2002-06-28 Procede de reduction enantioselectif d'une cetone aromatique prochirale comprenant au moins un groupe trifluoromethyle sur le cycle aromatique

Publications (1)

Publication Number Publication Date
EP1409704A2 true EP1409704A2 (de) 2004-04-21

Family

ID=8865018

Family Applications (1)

Application Number Title Priority Date Filing Date
EP02758523A Withdrawn EP1409704A2 (de) 2001-07-02 2002-06-28 Verfahren zur enantioselektiven reduktion von prochirale ketoverbindungen die mindestens ein trifluormethylverbindung auf dem aromatischen zyklus haben

Country Status (7)

Country Link
US (1) US20040191880A1 (de)
EP (1) EP1409704A2 (de)
JP (1) JP2004533269A (de)
AU (1) AU2002324106A1 (de)
CA (1) CA2452263A1 (de)
FR (1) FR2826650A1 (de)
WO (1) WO2003004666A2 (de)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2005317189A1 (en) * 2004-12-16 2006-06-22 Merck & Co., Inc. Process for the synthesis of (S)-1-(3,5-bis(trifluoromethyl)-phenyl)ethan-1-ol

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE4014573C1 (de) * 1990-05-07 1991-10-10 Forschungszentrum Juelich Gmbh, 5170 Juelich, De

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO03004666A2 *

Also Published As

Publication number Publication date
WO2003004666A2 (fr) 2003-01-16
AU2002324106A1 (en) 2003-01-21
WO2003004666A3 (fr) 2004-02-19
CA2452263A1 (fr) 2003-01-16
US20040191880A1 (en) 2004-09-30
JP2004533269A (ja) 2004-11-04
FR2826650A1 (fr) 2003-01-03

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