EP1108053A1

EP1108053A1 - Method for preparing (2r)-piperidine derivatives

Info

Publication number: EP1108053A1
Application number: EP99944524A
Authority: EP
Inventors: Michael Petersen; Wolfgang Beck; Klaus Heinzmann
Original assignee: Lonza AG
Current assignee: Lonza AG
Priority date: 1998-08-26
Filing date: 1999-08-26
Publication date: 2001-06-20
Also published as: CA2341554A1; JP2002523107A; WO2000012745A1; AU5741599A

Abstract

The invention relates to a method for preparing (R)-piperidine derivatives of the general formulas (I) and (VII), where R1 is hydroxy, amino or C¿1-6?-alkoxy and R?4¿ is hydroxy or amino. Said (R)-piperidine derivatives are obtained from (RS)-piperidine derivatives of the general formula (II), in which R2 is hydroxy, amino or C¿1-6?-alkoxy, in the presence of trace elements and using micro-organisms or cell-free enzymes from said micro-organisms.

Description

Process for the preparation of {2R) -piperidine derivatives

description

The present invention relates to a new process for the preparation of (2R) - piperidine derivatives of the general formulas

and

wherein R ^{1 is} hydroxy, amino or C ,. -AIkoxy and R ^{4 is} hydroxy or amino

(2R) -piperidine derivatives of the general formula I, such as e.g. (R) -Pipecolinic acid, are important synthesis building blocks for biologically active compounds, such as stimulants of growth hormone release (WO-A-9513069) or anti-anxiety agents (DE-A-37 02 943).

EP-A-686 698 describes a biotechnological process for the production of S-α-aminocarboxylic acids, for example S-α-pipecolinic acid, by reacting the racemic RS-α-aminocarboxylic acid amides with microorganisms of the genera Klebsiella and Pseudomonas, and also the corresponding ones R-α-aminocarboxylic acid amides are obtained In order to prepare the R-α-aminocarboxylic acid amides, however, the S-α-aminocarboxylic acids have to be separated off using relatively complex processes

A biotechnological process for producing (R) -pipecolinic acid by cultivating microorganisms of the genus Alcaligenes in (RS) -pipecolinic acid is also known. containing medium (Mochizuki. K. Yamazaki. Y. Maeda, H. Agric Biol Chem, 1988, 52 (5), 1 1 13) This method has the disadvantage that the (R) -pipeeolinic acid mixed with (S) - 2-aminoadipic acid is obtained and separated from the amino adipic acid formed after the preparation m ss

A process for the preparation of (R) -pipecolinic acid esters is described by RJ Kazlauskas et al (J Org Chem, 1994, 59, 2075-2081). The corresponding racemic esters are converted to the desired product by means of isolated stereoselective lipases from Aspergillus niger This method is that the desired products are in poor enantiomeric purity and are obtained in low yield. Furthermore, very large amounts of enzyme are also necessary

JP-A-63 248 393 describes a process for the preparation of (R) -pipecolinic acid starting from racemic pipecolinic acid by means of microorganisms of the genera Pseudomonas, Kurthia or Alcaligenes. The long reaction times are disadvantageous in this process

The object of the present invention is to eliminate these disadvantages and to provide a simple process for the preparation of (2R) -piperidine derivatives, with which high yields can be achieved and which is feasible on an industrial scale

This task is solved with the new biotechnological method according to claim 1

According to the invention, microorganisms are used which have the property of α-aminocarboxamides in foam of racemate or its optically active isomers of the general formula III,

wherein A together with -NH and -CH represent an optionally substituted 5- or 6-membered saturated heterocyclic ring as the only nitrogen source that can be used. Such microorganisms are known in the prior art and have already been described in detail in EP-A-686 698 Preferred microorganisms with the properties described above are microorganisms of the genera Klebsiella and Pseudomonas. Microorganisms of the species Pseudomonas putida, in particular those of the species Pseudomonas putida with the designation DSM 9923, and their functionally equivalent variants and mutants are particularly preferred

The microorganisms of the species Pseudomonas putida with the designation DSM 9923 were on 20 04 1 95 at the German Collection for Microorganisms and Cell Cultures GmbH (DSM). Mascherodeweg lb, D-38124 Braunschweig, deposited in accordance with the Budapest Treaty

^" Functionally equivalent variants and mutants" are understood to mean microorganisms which have essentially the same properties and functions as the original microorganisms. Such variants and mutants can be formed accidentally, for example by UV irradiation

Surprisingly, it has been found that these microorganisms, or enzyme-free enzymes therefrom, the S isomer in (RS) -piperidine derivatives of the general formula II

where R "is hydroxyl, amino or d .c-alkoxy, can decompose in the presence of trace elements and under aerobic conditions with ring cleavage and, in the case of microorganisms, can be used as the only source of carbon and energy. After a sufficient reaction time, usually after 1 to 48 hours, then there is little or no ring-shaped S-α-aminocarboxylic acid or other disruptive open-chain by-products such as S-2-aminoadipic acid in the reaction medium in the reaction medium. The (2R) -piperidine derivatives of the general formula I accumulated during the reaction

wherein R is hydroxy, amino or C |. _r , -alkoxy means can then be isolated without difficulty, in particular without complex separation processes

C] _{- f} , -Aikoxy has the meaning of a straight-chain or branched alkoxy group with 1 to 6 C atoms here and below. Methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy should be mentioned. tert-butoxy, pentoxy and its isomers and hexoxy and its isomers

The enzymes for the cell-free system can be obtained by disrupting the microorganisms in a manner customary in the art. For example, the ultrasound, French press or lysozyme method can be used for this purpose. These cell-free enzymes can also be immobilized on a suitable carrier material

Among trace elements are, for example, zinc, manganese, cobalt, copper, nickel, molybdenum, calcium, magnesium, boi. To understand iron or sulfur, the optimal composition of the trace elements depending on the microorganism used. Zinc and manganese are preferred. Cobalt, copper, nickel, molybdenum, calcium, magnesium. Boron, iron and sulfur »emeinsa 'sge" sets

The reaction of (RS) -piperidine derivatives of the general formula II is expediently carried out with growing microorganisms

The most preferred substrates of the general formula II are (RS) -pipecolinic acid, (RS) -pipecolinic acid ethyl ester, (RS) -pipecolinic acid isopropyl ester and (RS) -pipecolinic acid amide

Aerobic conditions are understood to mean culture conditions in which the microorganisms are supplied with oxygen. This can be experienced, for example, in shake culture by atmospheric oxygen or in submerged culture by blowing in molecular oxygen or atmospheric oxygen The reaction can be carried out without conventional cultivation directly by adding the microorganisms to (RS) -piperidine derivatives of the general formula II. Alternatively, the reaction can be carried out after conventional cultivation of the microorganisms with a suitable carbon and energy source Sugar. Carboxylic acids, sugar alcohols or amino acids can be used. As sugars, hexoses such as glucose or pentoses can be used For example, glutamate can be used

The reaction is preferably carried out without customary cultivation directly by adding the microorganisms to (RS) -piperidine derivatives of the general Foimel II

During the reaction, the microorganisms can use the (S) -isomer of the (RS) -piperidine derivative of the general formula II as the only carbon and energy source and as the only nitrogen source

The medium customary in the art can be used as the medium for the process according to the invention, such as mineral salt media, for example the mineral salt medium according to Kulla et al (Arch Microbiol 135, 1-7. 1983), the media described in Table 1 or low-molar buffers such as 10 to 100 mM phosphate buffer to which the necessary trace elements have been added. The process is preferably carried out in the media according to Table 1

The conditions are expediently chosen such that the S-isomer of the (RS) -piperidine derivatives of the formula II is preferably used by the microorganisms as a carbon and energy source. The medium therefore preferably does not contain any compounds other than compound II that are easily usable as a carbon and energy source links

The reaction is carried out expediently with a single, repeated or continuous addition of (RS) -piperidine derivatives of the general formula II. The addition of (RS) -piperidine derivatives of the general formula II is carried out in such a way that the concentration is 30% by weight, preferably 10% by weight %, particularly preferably 5% by weight, not exceeded The pH of the medium is expediently in a range from 4 to 9, preferably from 6 to 8 The reaction is expedient at a temperature of from 15 to 80 ° C., preferably from 25 to 40 ° C. by 'saw ¹ -f

Which end product of the general formula I is obtained in the reaction depends on the choice of the starting material and the reaction conditions, in particular the reaction time and the oxygen supply

Starting from (RS) -pipecolinic acid [(RS) -piperidine derivative II, in which R ^ means hydroxy], only the (2R) -pipecolic acid can be obtained Starting from (RS) -pipecolinic acid amide [(RS) -piperidine derivative II, in which R ^ Amino means], both the (2R) -pipecolinic acid amide and the (2R) -pipecolinic acid can be obtained starting from an (RS) -pipecolinic acid ester [(RS) -piperidine derivative II, where R ^ C] _5-alkoxy] both the (2R) -pipecolinic acid ester and the (2R) -pipecolinic acid are obtained

Whether (2R) -pipecolinic acid amide or (2R) -pipecolinic acid is obtained from (RS) -pipecolinic acid amide can be controlled, for example, via the reaction time. Thus, when (RS) -pipecolinic acid amide is reacted with the microorganisms according to the invention, the (2R) initially Pipecolinic acid amide, which can be isolated if necessary After a longer reaction time, the acid is formed from the (2R) -pipecolinic acid amide. The reaction can be followed analytically, e.g. chromatographically. The same applies to the reaction of (RS) -pipecolinic acid esters, in which the (2R ) -Pipecolinic acid ester and then the acid accumulates

After a customary reaction time of preferably 1 to 48 hours, the (2R) -piperidine derivatives of the general formula I, preferably the (R) -pipecolinic acid, the (R) -pipecolinic acid amide and the (R) -pipecolinic acid ethyl and isopropyl esters, become very high to quantitative Yield obtained in the medium

The (2R) -piperidine derivatives of the general formula I obtained in this way can be isolated by customary work-up methods such as, for example, by separating off the biomass, acidification, chromatography, electrodialysis or crystallization

Alternatively, the course of the reaction can also be regulated via the supply of oxygen. If, for example, (RS) -pipecolinic acid amide or (RS) -pipecolinic acid ester (R) -pipecolinic acid are to be produced, the reaction is expedient until the (R) -pipecolinic acid amide or the like is accumulated (R) -pipecolinic acid esters as above described carried out under aerobic conditions and then the oxygenation adjusted, after which the acid is formed

The implementation of the (RS) -pipecolinic acid derivative according to formula V

wherein R ^{1 is} amino or C]., - alkoxy, where Cι.r, -alkoxy has the meaning given above, for (R) -pipecolinic acid according to formula IV

in effect via the (R) -pipecolinic acid derivative according to formula VI,

where R ^"has the meaning given above, which can be temporarily accumulated in the medium with increasing enantiomeric excess and, if appropriate, isolated. If the (R) -pipecolinic acid derivative according to formula VI is to be prepared from the (RS) -pipecolinic acid derivative according to formula V, it will after accumulation isolated If the (R) -pipecolinic acid according to formula IV is to be produced from the (RS) -pipecolinic acid derivative according to formula V, the reaction is expediently carried out under aerobic conditions until the accumulation of the (R) -pipecolinic acid derivative according to formula VI and then the oxygen supply is stopped

To determine whether the (R) -pipecolinic acid derivative according to formula VI is accumulated in the medium, the reaction can be followed analytically, for example by chromatography Implementation should be stopped as soon as the accumulation of the (R) -pιpecolιnsauredeπ \ ats according to formula VI is reached

Another component of the invention is the further implementation, the reduction, of the (2R) - Pipecolmsauredeπvate dei general formula VI to the (2R) -Pιpeπdιndeπ \ aten of the general Foimel VII

where R is Hvdroxv or Amino

Usually, an alkali metal hydride or an alkaline earth metal hydride such as, for example, is used as the reducing agent Potassium or

Natπumaluminiumhydrid Magnesium- or Calciumborhydπd used In particular, Lithiumaluminiumhydπd

The reduction is expediently carried out at a temperature of 0 to 100 ° C., preferably at the reflux temperature of the corresponding solvent

As is known in the art, the reduction can be carried out in a polar organic solvent, for example in an ethei. Diethyl ether, dipropylethei, tetrahydrofuran or 1,4-dioxane are suitable as ethers, for example

After a customary reaction time of 1 to 16 h, the desired products of the general formula VII, such as the (R) -2- (aminomethyl) p; perιdιn or the (R) -2- (hydroxymethyl) pιpeπdιn, can then be isolated by known processing methods Table 1

Medium 1

(NH.,) ₂ SO ₄ 2.0 g / 1

Na ₂ HPO ₄ 2.0 g / 1

KH ₂ PO 1, 0 g / 1

NaCl 2.0 g / 1

MgCl ₂ • 6 H ₂ O 0.4 g / 1

CaCl ₂ • 2 H ₂ O 14.5 mg / 1

FeCl ₃ • 6 H ₂ O 0.8 mg / 1

ZnSO., • 7 H ₂ O 0.1 mg / 1

MnCl ₂ • 4 H ₂ O 0.09 mg / 1

H ₃ BO ₃ 0.3 mg / 1

CoCl ₂ ^• 6 H ₂ O 0.2 mg / 1

CuCl ₂ • 2 H ₂ O 0.01 mg / 1

NiCl ₂ • 6 H ₂ O 0.02 mg / 1

Na ₂ MoO ₄ • 2 H ₂ O 0.03 mg / 1

FeSO ₄ • 7 H ₂ O 2.0 mg / 1

EDTA-Na ₂ • 2 H ₂ O 5.0 mg / 1

Medium 2

KH ₂ PO 1, 3 g / 1

NH ₄ C1 5 g / 1

(NH) ₂ SO ₄ 2 g / 1

Na ₂ SO ₄ 0.25 g / 1

MgCl ₂ • 6 H ₂ O 0.8 g / 1

CaCl ₂ • 2 H ₂ O 0.16 g / 1

ZnSO ₄ • 7 H ₂ O 9 mg / 1

MnCl ₂ ^• 4 H ₂ O 4 mg / 1

H ₃ BO ₃ 2.7 mg / 1

CoCl ₂ • 6 H ₂ O 1.8 mg / 1

CuCI ₂ ^• 2 H ₂ O 1.5 mg / 1

NiCl ₂ • 6 H ₂ O 0. 18 mg / 1

Na ₂ MoO., 2 H ₂ O 0.2 mg / 1

FeSO ₄ • 7 H ₂ O 30 mg / 1

EDTA-Na ₂ • 2 H ₂ O 175 mg / 1 Me ium 3

Na _: SO., 0.1 g / 1

Na ₂ HPO, 2.0 g / 1

KH ₂ PO ₄ 1.0 g / 1

NaCl 2.0 g / 1

MgCl ₂ • 6 H ₂ O 0.4 g / 1

CaCl ₂ ^• 2 H ₂ O 14.5 mg / 1

FeCl ₃ • 6 H ₂ O 0.8 mg / 1

ZnSO • 7 H ₂ O 0.1 mg / 1

MnCl ₂ • 4 H ₂ O 0.09 mg / 1

H ₃ BO ₃ 0.3 mg / 1

CoCl ₂ • 6 H ₂ O 0.2 mg / 1

CuCl ₂ ^• 2 H ₂ O 0.01 mg / 1

NiCl ₂ • 6 H ₂ O 0.02 mg / 1

Na ₂ MoO ₄ • 2 H _: O 0.03 mg / 1

FeSO., ^■ 7 H ₂ O 2.0 mg / 1

EDTA-Na ₂ • 2 H ₂ O 5.0 mg / 1

Examples

example 1

Production of (R) -Pipecoiinsäιιre from (RS) -Pipecoiinsäure

1.1 in Schütteikuitur

400 ml of medium 1 with a concentration of 1.5% (RS) -pipecolinic acid were placed in a 1-1 Erlenmeyer flask and mixed with Pseudomonas putida DSM 9923. The mixture was incubated at 30 ° C. and 140 revolutions per minute after For 24 hours, the cell-free supernatant was examined for the content of (R) -pipecolinic acid at an OD650 of 3.7 using HPLC. (R) -Pipecolinic acid was in a yield of> 45% based on the (RS) -pipecolinic acid used and with a Obtained ee value of> 99%

1.2 in the fermenter 100 ml of medium 3 with a concentration of 1% (RS) -pipecolinic acid were placed in a 500 l Erlenmeyer flask with baffles and mixed with Pseudomonas putida DSM 9923. The mixture was incubated at 30 ° C. and 140 revolutions / min on a shaking machine for 16 h. 3 l of medium 2 with a concentration of 1.5% (RS) -pipecolinic acid (45 g) were placed in a 5 l fermenter and mixed with the Preculture of Pseudomonas putida DSM 9923 added After 7 h, a further 45 g of (RS) -pipescolic acid were added continuously as a 10% solution within 5 h. After a further 4 h, the biotransformation was stopped and the cell-free supernatant was determined by HPLC to determine the (R) Pipecolinic acid examined (R) - Pipecolinic acid was obtained in quantitative yield and with an ee value of> 97%

Example 2

Production of (R) -pipecolic acid from (RS) -pipecoic acid ethyl ester

40 ml of medium 3 with a concentration of 1.5% (RS) -pipecolinic acid ethyl ester were placed in a 100 ml Erlenmeyer flask with baffles and mixed with Pseudomonas putida DSM 9923. The mixture was at 30 ° C. and 140 revolutions / min on a

Incubated shaking machine After 26 h at an OD650 of 2.0, the cell-free supernatant was examined with HPLC for the content of (R) -pipecolinic acid (R) -pipecolinic acid was obtained in> 45% yield based on the (RS) -pipecolinic acid ethyl ester used and obtained with an ee value of> 80%. After 44 h, at an OD650 of 3.2, enantiomerically pure (R) -pipecolinic acid was obtained Example 3

Preparation of (R) -pipecolic acid amide from (RS) -pipeeolic acid amide in shake culture

40 ml of medium 3 with a concentration of 1.5% (RS) -pipecolinic acid amide were placed in a 100 ml Erlenmeyer flask with baffles and mixed with Pseudomonas putida DSM 9923. The mixture was stirred at 30 ° C. and 140 rpm on a shaker incubated After 20 h at an OD650 of 3.0 the cell-free supernatant was examined for the content of (R) -pipecolinic acid amide with HPLC (R) -pipecolinic acid amide was obtained in a yield of> 45% based on the (RS) -pipecolinic acid amide and obtained with an ee value of> 99%

Example 4

Production of (R) -Pipecolinsäιιreamid and (R) -Pipecolinsäure from (RS) - Pipecoiinsäurcamid in the fermenter

Pseudomonas putida DSM 9923 was grown on medium 3 with 20 g / l glutamate overnight in the fermenter. At OD650 of 14, 1% (RS) -pipecolinic acid amide was added. According to GC analysis, after 2 hours there was practically enantiomerically pure (R) -pipecolinic acid amide before pipecolinic acid was not to prove

A sample of this solution was incubated in the shaker cabinet with the exclusion of air at 37 ° C. and 140 revolutions / min. According to GC analysis, enantiomerically pure (R) -pipecolinic acid before pipecolinic acid amide was no longer detectable after 48 h

Example 5

Production of (R) -pipecolic acid from (RS) -pipecoic acid amide in shake culture 40 ml of medium 3 with a concentration of 1% (RS) -pipecolinic acid amide were presented in a 100 ml Erlenmeyer flask with baffles and mixed with Pseudomonas putida DSM 9923 was incubated at 30 ° C. and 140 revolutions / min on a shaking machine. After 26 h the cell-free supernatant was examined by GC at an OD650 of 3 9. (R) -pipecolinic acid with ee> 98% was found in approx. 10% yield Example 6

Preparation of (R) -pipecolinic acid isopropyl ester from (RS) -pipecolic acid isopropyl ester in bulk with 100 ml of minimal medium 3 with a concentration of 4 g / l of (RS) -pipecolinic acid isopropyl ester was presented with baffles in a 500 ml Erlenmeyer flask and treated with Pseud23omonas putida DSM The mixture was incubated at 30 ° C. and 140 rpm on a shaking machine. After 4.5 h at an OD650 of 1.4, the cell-free supernatant was examined with GC for the content and enantiomeric excess of pipecolic acid and pipopropolic acid isopropyl ester (R) -pipecolinic acid isopropyl ester was enamino pure and obtained in a yield of 88% based on the isopropyl (R) -pipecolinate used A few percent of enantiomerically pure (R) -pipecolinic acid were found as a by-product

After separating the biomass by centrifugation (20 min at 8000 ^), the aqueous solution was evaporated to a third in vacuo, adjusted to pH 10 with 30% sodium hydroxide solution and immediately extracted three times with 20 ml of diethyl ether. The combined organic phases were dried with sodium sulfate and concentrated 0.195 g of (R) -pipecolinic acid isopropyl ester was crystallized as hydrochloride by introducing hydrochloric acid (80% yield, ee> 98% according to GC)

Example 7

Preparation of (R) -pipecolic acid from (RS) -pipecolic acid isopropyiester 100 ml minimal medium 3 with a concentration of 4 g / 1 (RS) -pιpecolinic acid; sopropyl esters were placed in a 500 ml Erlenmeyer flask with baffles and with Pseudomonas putida DSM °> 923 added. The batch was incubated at 30 ° C. and 140 rpm on a shaking machine. After 7.5 h, a sample was taken at an OD650 of 1.4 and incubated with exclusion of air. After 22 h, the cell-free supernatant was checked for content and enantiomeric excess by GC investigated on pipecolinic acid and pipecolinic acid isopropyl ester (R) -pipecolinic acid was obtained enantiomerically pure and in a yield of 80% based on the isopropyl ester of (R) -pipecolinic acid as a by-product, a few percent of enantiomerically pure (R) -pipecolinic acid isopropyl ester was found Example 8

Reduction of (R) -pipecoliιιsäureamid to (R) -2- (aminomethyl) piperidine

In a 2-1 flask, 1 1.8 g of lithium aluminum hydride were suspended in 500 ml of diethyl ether under a nitrogen atmosphere. At 5 ° C., 20 g of (R) -pipecolinic acid amide (see Example 3) were slowly added. The mixture was stirred for 3 hours at room temperature and then for 5 hours stirred under reflux After cooling to room temperature, 12 ml of water, 12 ml of 1% sodium hydroxide solution and 36 ml of water were slowly added dropwise in succession. The mixture was stirred at room temperature until it was snow-white. 120 g of sodium sulfate were added and the mixture was fed at room temperature overnight. The solid was filtered off and washed four times with 250 ml of diethyl ether each time. The combined organic phases were evaporated in vacuo. 12.5 g of (R) -2- (aminomethyl) piperidine were obtained as yellowish oil (66% according to GC) distillation at 52-53 ° C / 18 mbar gave 3 6g (20%) GC-pure (R) -2- (aminomethyl) piperidine as a colorless liquid

Example 9

Reduction of (R) -pipecolinic acid isopropyl ester to (R) -2- (hydroxymethyl) -piperidine In a 250 ml flask, 0.2 g of lithium aluminum hydride were suspended in 20 ml of diethyl ether under a nitrogen atmosphere. At 5 ° C, 90 mg of (R) -pipecolinic acid isopropyl ester were suspended - hydrochloride slowly added. The mixture was stirred at room temperature for 1 h and then under reflux for 4 h. After cooling to room temperature, 0.2 ml of water were slowly added in succession. 0.2 ml of 15% sodium hydroxide solution and 0.4 ml of water were added dropwise. The mixture was stirred at room temperature until it was snow white. 2 g of sodium sulfate were added and the mixture was stirred at room temperature overnight. The solid was filtered off and washed four times with 20 ml of diethyl ether. The combined Organic phases were evaporated in vacuo. 47 mg of (R) -2- (hydroxymethyl) piperidine were obtained as a slightly yellowish oil (content 95%, ee> 97% according to GC)

Claims

claims

Process for the preparation of (2R) - piperidine derivatives of the general formula

wherein R is hydroxy, amino or C]. _ή -A] koxy means, characterized in that an (RS) - piperidine derivative of the general formula

1 1 1 * wherein R is hydroxy, amino or Cι _ <-, - alkoxy and R and R may be the same or different with the restriction that when R is amino R is not hydroxy or Cι_ _f , -alkoxy and when R ^{1 is} C ]. _f) -alkoxy is R is not hydroxy or amino, with microorganisms that are capable, α-aminocarboxamides, in the form of the racemate or one of its optically active isomers, the general formula

wherein A together with -NH and -CH represents an optionally substituted 5- or 6-membered saturated heterocyclic ring, to be used as the sole nitrogen source, or cell-free enzymes from these microorganisms, in the presence of trace elements and under aerobic conditions, the S -Isomers of the (RS) -piperidine derivative II is used or decomposed as a carbon and energy source with ring cleavage. Method according to claim I. characterized in that the substrate of the formula II (RS) -pipecolinic acid. (RS) -PψecolinsaureethyΙester, (RS) -Pipecolinsaureisopropylester or (RS) -Pιpecolinsaureamid used

Method according to one of claims 1 or 2, characterized in that the reaction with microorganisms of the genus Pseudomonas, in particular the species Pseudomonas putida, is carried out.

Method according to one of claims 1 to 3, characterized in that the reaction with microorganisms of the species Pseudomona putida (DSM 9923) or their functionally equivalent variants and mutants is carried out

Method according to one of claims 1 to 4, characterized in that the reaction is carried out with growing microorganisms

Method according to one of claims 1 to 5, characterized in that the addition of (RS) - pipecolinic acid derivatives of the general formula II is carried out in such a way that the concentration does not exceed 30% by weight

Method according to one of claims 1 to 6, characterized in that the reaction is carried out at a pH of 4 to 9 and a temperature of 15 to 80 ° C.

Process for the preparation of (R) -pipecolinic acid of the formula

characterized in that (RS) -pipecolinic acid derivatives of the general formula

wherein R ^{1 is} amino or C | _-f , -alkoxy means, as a substrate with microorganisms which are capable, α-aminocaibonic acid amides, in the form of the racemate or one of its optically active isomers, of the general formula

wherein A together with -NH and -CH represent an optionally substituted 5- or 6-membered saturated heterocyclic ring, to be used as the sole nitrogen source, or by means of cell-free enzymes from these microorganisms, in the presence of trace elements and under aerobic conditions, the reaction until the accumulation of the corresponding (R) -pipecolinic acid derivative of the formula

wherein R has the meaning given, which can optionally be isolated, is carried out under aerobic conditions, the S-isomer of the (RS) -piperidine derivative II being utilized or decomposed as a source of carbon and energy with ring cleavage, and the oxygen supply is then discontinued

Process for the preparation of (2R) -piperidine derivatives of the general formula

wherein R ^{4 is} hydroxy or amino, characterized in that in the first step (RS) -Pipecolinsauredeπvate of the general formula

wherein R ^{1 is} amino or Cι. ₆ - λlkoxy means, as a substrate with microorganisms which are capable, α-aminocarboxamides, in the form of the racemate or one of its optically active isomers, of the general formula

wherein A together with -NH and -CH means an optionally substituted 5- or 6-membered saturated heterocychic ring, to be used as the sole nitrogen source, or by means of cell-free enzymes from these microorganisms, in the presence of trace elements and under aerobic conditions under which the S -Isomers of the (RS) -piperidine derivative II with ring cleavage as a carbon and energy source is used or degraded, into a (R) -pipecolinic acid derivative of the general formula

wherein R is amino or Cι. _f , -alkoxy means, and this reduced in the second stage into the desired compound of general formula VII