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WO2000050628A1 - Hydrolyse enzymatique enantioselective d'esters 3-substitue d'acide glutarique - Google Patents

Hydrolyse enzymatique enantioselective d'esters 3-substitue d'acide glutarique Download PDF

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Publication number
WO2000050628A1
WO2000050628A1 PCT/US2000/004668 US0004668W WO0050628A1 WO 2000050628 A1 WO2000050628 A1 WO 2000050628A1 US 0004668 W US0004668 W US 0004668W WO 0050628 A1 WO0050628 A1 WO 0050628A1
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WIPO (PCT)
Prior art keywords
enantiomeric excess
enantiomer
hydrolysis
formula
carried out
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.)
Ceased
Application number
PCT/US2000/004668
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English (en)
Inventor
Michael J. Homann
William B. Morgan
Aleksey Zaks
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.)
Merck Sharp and Dohme LLC
Original Assignee
Schering Corp
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Filing date
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Priority to AU35005/00A priority Critical patent/AU3500500A/en
Publication of WO2000050628A1 publication Critical patent/WO2000050628A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • 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
    • C12P41/00—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture
    • C12P41/003—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by ester formation, lactone formation or the inverse reactions
    • C12P41/005—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by ester formation, lactone formation or the inverse reactions by esterification of carboxylic acid groups in the enantiomers or the inverse reaction
    • 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
    • C12P7/62—Carboxylic acid esters

Definitions

  • Substituted oximes are useful pharmaceutical compounds.
  • International Application No. PCT/US98/23255 filed November 18, 1998, describes certain substituted oximes that are useful as neurokinin antagonists.
  • it is desireable to produce a particular enantiomer of the substituted oxime.
  • any efficient enantioselective process for producing a chiral intermediate for these compounds would be a welcome contribution to the art. This invention provides such a contribution.
  • This invention provides a process for preparing an S-enantiomer compound having the formula
  • alkyl means straight or branched hydrocarbon chains of 1 to 6 carbon atoms, optionally substituted with one or more halo, hydroxy, or alkoxy substituents.
  • Alkoxy refers to a group having the formula R-O-, wherein R is alkyl.
  • Aryl refers to a carbocyclic group having at least one aromatic ring (e.g., phenyl or naphthyl), optionally substituted with one or more substituents selected from halo, alkyl, hydroxy, alkoxy, or -CF3.
  • Alkyl refers to a group having the formula aryl-R-, wherein R is alkyl.
  • Cycloalkyl refers to a non-aromatic carbocyclic ring of from 3 to 6 carbon atoms, optionally substituted with one or more substituents selected from halo, alkyl, hydroxy, alkoxy, or -CF3.
  • Cycloalkylalkyl refers to a group having the formula cycloalkyl-R-, wherein R is alkyl.
  • Halo refers to fluorine, chlorine, bromine or iodine.
  • Et refers to an ethyl group.
  • Enantiomeric excess is calculated according to the following formula:
  • R 1 is preferably alkyl, more preferably methyl or ethyl, most preferably ethyl.
  • Enzymes suitable for use in the present process can be identified by carrying out the screening procedure described in Example 1, below.
  • the enzyme is preferably one that is capable of producing an e.e. of the desired compound of at least 80%, more preferably at least 90%.
  • the enzyme is one that produces the S-enantiomer compound in enantiomeric excess.
  • the enzymes used for preparing the S-enantiomer compound of formula (IA) are produced by Candida rugosa , Candida cylindracea , Candida antarctica , and Rhizopus delemar .
  • Examples of enzymes suitable for use in preparing the S-enantiomer compound of formula (IA) include, but are not limited to, the following commercially available enzyme preparations: Altus ChiroCLEC CRO (Candida rugosa); Altus ChiroCLEC-CR (Candida rugosa); Altus Lipase CR Analytical Grade 001-C (Candida rugosa); Biocatalysts Ltd.
  • Candida cylindracea Meito Sangyo LIPASE-OF (Candida cylindracea); Boehringer Mannheim Cholesterol Esterase (Candida rugosa); Boehringer Mannheim CHIRAZYME L-2 (Candida antarctica, fraction B); Fluka Lipase (Candida antarctica); Genzyme Lipase (Candida cyclindracea); Novo Nordisk Novozym 435 (Candida antarctica, type B); Novo Nordisk SP 525 (Candida antarctica, type B); and Seikagaki Lipase (Rhizopus delemar).
  • Boehringer Mannheim CHIRAZYME L-2 in either the dry or liquid form
  • Novo Nordisk Novozym 435 in either the dry or liquid form
  • the enzymes used for preparing the R-enantiomer compound of formula (IB) are obtained from porcine or bovine pancreas.
  • enzymes suitable for use in preparing the R-enantiomer compound of formula (IB) include, but are not limited to, the following commercially available enzyme preparations: Biocatalysts Ltd. Lipase (porcine pancreas); Boehringer Mannheim Lipase (porcine pancreas); Boehringer Mannheim CHIRAZYME L-7 Lipase (porcine pancreas); Rohm Tech COROLASE PP (porcine pancreas); Scientific Protein Labs.
  • ThermoCat 027 Of these commercially available enzyme preparations, Sigma ⁇ - Chymotrypsin Type II and Boehringer Mannheim CHIRAZYME L-7 are particularly preferred.
  • the hydrolysis of compound (II) is preferably carried out at a pH of 5-9, more preferably 6 - 8.5, most preferably 7-8.
  • the substrate concentration is 5% to 25%, more preferably 8% to 12%.
  • the hydrolysis is carried out at a temperature of 25° to 45° C, more preferably, 30° to 40° C.
  • the hydrolysis is preferably carried out in the presence of a buffer (e.g., sodium phosphate or potassium phosphate buffer), and the reaction may, if desired, be titrated with a caustic titrant (e.g., NaOH solution) to maintain the pH within the preferred range.
  • a buffer e.g., sodium phosphate or potassium phosphate buffer
  • a caustic titrant e.g., NaOH solution
  • the concentration of the caustic titrant is 0.2 to 1 M, preferably about 0.5 M.
  • compound (II) is dispersed in a buffer solution by agitation, and the enzyme is subsequently added to the mixture. Upon adding the enzyme, the reaction mixture is agitated until the desired degree of hydrolysis is reached.
  • an enzyme in solid form e.g., adsorbed on beads
  • the reaction may be terminated by removing the enzyme by filtration, and adding acid (e.g., H 2 SO 4 ) to the filtrate to adjust the pH to about 4.0- 4.5, thereby precipitating the desired product.
  • acid e.g., H 2 SO 4
  • the reaction may be terminated by adding acid (e.g., H 2 SO 4 ) to adjust the pH to about 4.0-4.5, and the precipitated product can be recovered by filtration.
  • Compound (II) may be made by conventional means, e.g., as shown in the scheme below: CH 3 COCH 2 COOEt
  • aldehyde (1) is condensed with ethyl acetoacetate in ethyl alcohol with a piperidine catalyst to form crude reaction product (2), a mixture of 2 isomers.
  • the condensation is preferably carried out over 2-3 days at 20°C ⁇ 5°C.
  • Potassium hydroxide is added to the crude reaction product (2), and heated at about 60°-70°C for about one hour to form dipotassium salt (3), which is filtered and washed with ethyl alcohol.
  • the dipotassium salt is dissolved in water and treated with HC1 to form compound (4).
  • Compound (4) is reacted with an alcohol, R ⁇ H (e.g., ethyl alcohol) in the presence of p-toluenesulfonic acid to form compound (II).
  • R ⁇ H e.g., ethyl alcohol
  • Compound (II) may be isolated and purified by adding a higher boiling solvent, (e.g., heptane), distilling off the alcohol, washing with dilute sodium bicarbonate solution to remove the p-toluenesulfonic acid, and cooling the solution to precipitate compound (II).
  • a higher boiling solvent e.g., heptane
  • Enzyme screening reactions were conducted using 1-30 mg enzyme suspended in 0.9 ml of 50 mM sodium phosphate buffer pH 7. The reaction was initiated upon addition of ethyl 3-[3',4'-dichlorophenyl]glutarate (-25 mg) dissolved in 0J ml acetone (10% v/v). Following 48 hours of incubation in 1 dram vials at 30° C with agitation (225 rpm), the reactions were terminated by acidification to a pH of less than 2 using 6N HC1 and extracted with 2 ml ethyl acetate prior to analysis by reverse-phase and chiral HPLC. Enzymes generating the R -monoester or the S -monoester in > 87% enantiomeric excess are summarized in Tables 1 and 2, respectively.
  • Ethyl 3-(3',4'-dichlorophenyl)glutarate (2.45 g, 7.35 mmol) was added to 50 mM KC1 (400 mL) in a 3-neck round bottom flask equipped with an electrode and pH delivery tube connected to a pH stat. The mixture was stirred and adjusted to pH 7-8. ⁇ -chymotrypsin (Sigma Type II; bovine pancreas) (1.25 g) was added and the pH was maintained at pH 8 by automatic titration of 0.5 M NaOH. The reaction was terminated after 114 hours. The reaction mixture was evaporated to dryness and the resultant solid triturated with methyl ene chloride (200 mL).
  • Ethyl 3-[3',4'-dichlorophenyl]glutarate (135 g) was dispersed in 10 mM sodium phosphate buffer pH 8.0 (1215 ml) using an impeller mixer mounted in a 3-neck round bottom jacketed glass flask (3L) equipped with an electrode and pH delivery tube connected to a pH stat. The mixture was stirred and adjusted to pH 8 at 40° C. The reaction was initiated with the addition of 33.75 g of Chirazyme L-2 (adsorbed on beads) and pH 8.0 was maintained by automatic titration of 0.5 M sodium hydroxide. Following 24 hours of incubation, the reaction was 96%> monoester as measured by HPLC. The reaction was terminated by removing the enzyme by filtration.
  • the pH of the filtrate was adjusted to 4.0-4.5 using 20% H 2 SO 4 followed by isolation of the precipitated product by filtration.
  • the acid precipitant was dried (vacuum dryer), and then dissolved in 300 ml of tert -butyl methyl ether (TBME). Insoluble material was removed by filtration through celite.
  • the TBME filtrate was mixed with 400-600 ml of n-heptane and vacuum evaporated to remove the TBME. Removal of TBME resulted in precipitation of the desired S-monoester. Residual ethyl 3-[3',4'-dichlorophenyl]glutarate diester remains dissolved in n-heptane.
  • Multigram scale preparation of S-monoethyl ester using Chirazyme L-2 (liquid) Ethyl 3-[3',4'-dichlorophenyl]glutarate (20 g) was dispersed in 25 mM sodium phosphate buffer pH 7.5 (180 ml) using an impeller mixer mounted in a 3- neck round bottom jacketed glass flask (500 mL) equipped with an electrode and pH delivery tube connected to a pH stat. The mixture was stirred and adjusted to pH 7.5 at 30-38° C. The reaction was initiated with the addition of 10 ml of Chirazyme L-2 (liquid form) and pH 7.5 was maintained by automatic titration of 0.5 M sodium hydroxide.
  • Ethyl 3-[3',4'-dichlorophenyl]glutarate (50 kg) was dispersed in 10 mM sodium phosphate buffer pH - 8.0 ( 450 L) with agitation (impeller speed -100 rpm) at 40° C in a 300 gallon glass-lined reactor equipped with an in-vessel pH probe.
  • the reaction was initiated with the addition of 6.25 kg of Chirazyme L-2 (adsorbed on beads).
  • Control of reaction pH between 7.9 -8J was achieved manually by observing the in-vessel probe and manually adjusting a metering valve to regulate flow of caustic (0.5 M NaOH) from an adjacent feedbottle pressurized with 3 psi nitrogen.
  • a 93 % conversion yield was achieved at 38-40° C following 43 hours of incubation.
  • the reaction was terminated by removing the enzyme by filtration.
  • the enzyme filtercake was washed with 100 mM phosphate buffer pH -8.0 ( ⁇ 125L).
  • the filtercake wash and filtrate were combined in a 200 gallon reactor and the pH was adjusted to -4.2 using 10 % H 2 SO 4 followed by isolation of the precipitated product by filtration.
  • the acid precipitant was rinsed in situ with water (-100 L) and dried on trays in an air dryer, yielding 40.5 kg product in >99 % enantiomeric excess.
  • Dried cake from two batch reactions (-79 kg) was mixed with 6 kg of supercel and 320 L tert -butyl methyl ether (TBME).
  • Insoluble material and supercel were removed by filtration and rinsed with 110 L of TBME.
  • the rinse was combined with the filtrate and concentrated 2-3 fold by vacuum evaporation.
  • the TBME concentrate was mixed with - 640 L of n-heptane and vacuum evaporated to remove the TBME. Removal of TBME resulted in precipitation of the desired S-monoester. Residual ethyl 3-[3',4'- dichlorophenyl]glutarate remains dissolved in n-heptane.
  • Ethyl 3-[3',4'-dichlorophenyl]glutarate (6.5 kg) was dispersed in 25 mM sodium phosphate buffer pH - 8.0 ( -60 L) with agitation (impeller speed -100 rpm) at 30° C in a 50 gallon glass-lined reactor equiped with an in-vessel pH probe.
  • the reaction was initiated with the addition of 3.25 L of Chirazyme L-2 (liquid).
  • Control of reaction pH between 7.9 -8J was achieved manually by observing the in-vessel probe and manually adjusting a metering valve to regulate flow of caustic (0.5 M NaOH) from an adjacent portable can pressurized with nitrogen (20 psi).
  • the TBME concentrate was mixed with - 110 L of n-heptane and vacuum evaporated to remove the TBME. Removal of TBME resulted in precipitation of the desired S-monoester. Residual ethyl 3-[3',4'- dichlorophenyljglutarate remains dissolved in n-heptane. The precipitated slurry was filtered and the filtercake dried in a vacuum oven yielding a white powder; 9.7 kg, 81.4%) molar yield; > 99% enantiomeric excess.

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  • General Engineering & Computer Science (AREA)
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  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé de préparation d'un composé S-énantiomère possédant la formule (IA) avec un excès d'énantiomère ou un composé R-énantiomère possédant la formule (IB) avec un excès d'énantiomère, ledit procédé consistant à hydrolyser un composé possédant la formule (II) avec (a) une enzyme de produire un excès d'énantiomère du composé S-énantiomère ayant la formule (IA) d'au moins 70 % ou (b) une enzyme capable de produire un excès d'énantiomère du composé R-énantiomère ayant la formule (IB) d'au moins 70 %, R1 dans ladite formule étant sélectionné dans le groupe constitué d'alkyle, aralkyle cycloalkyle ou cycloalkylalkyle.
PCT/US2000/004668 1999-02-26 2000-02-24 Hydrolyse enzymatique enantioselective d'esters 3-substitue d'acide glutarique Ceased WO2000050628A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU35005/00A AU3500500A (en) 1999-02-26 2000-02-24 Enantioselective enzymatic hydrolysis of 3-substituted esters of glutaric acid

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US25947299A 1999-02-26 1999-02-26
US09/259,472 1999-02-26

Publications (1)

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WO2000050628A1 true WO2000050628A1 (fr) 2000-08-31

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AR (1) AR022725A1 (fr)
AU (1) AU3500500A (fr)
WO (1) WO2000050628A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007010356A3 (fr) * 2005-07-18 2007-08-23 Pfizer Ltd Procede pour la preparation de derives de sulfonamide

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3224019C1 (de) * 1982-06-28 1984-02-16 Takara Shuzo Co., Ltd., Kyoto Verfahren zur Herstellung von ß-(S)-Aminoglutarsäuremonoalkylestern
WO1993004058A1 (fr) * 1991-08-22 1993-03-04 Chiroscience Limited Esters de glutarate chiraux, leur resolution et composes de glutaramide derives
EP0812819A2 (fr) * 1996-06-10 1997-12-17 Hüls Aktiengesellschaft Enantiomère enrichie d'un ester de l'acide malonique substitué par des restes d'hydrocarbures tertiaires et sa préparation

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3224019C1 (de) * 1982-06-28 1984-02-16 Takara Shuzo Co., Ltd., Kyoto Verfahren zur Herstellung von ß-(S)-Aminoglutarsäuremonoalkylestern
WO1993004058A1 (fr) * 1991-08-22 1993-03-04 Chiroscience Limited Esters de glutarate chiraux, leur resolution et composes de glutaramide derives
EP0812819A2 (fr) * 1996-06-10 1997-12-17 Hüls Aktiengesellschaft Enantiomère enrichie d'un ester de l'acide malonique substitué par des restes d'hydrocarbures tertiaires et sa préparation

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
C.J. FRANCIS AND B. JONES: "Preparations of chiral delta-lactones via enantiotopically specific pig liver esterase-catalysed hydrolyses of 3-substituted glutaric acid diesters", J. CHEM SOC., CHEM COMMUN., no. 9, 1984, LONDON, UK, pages 579 - 580, XP002141862 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007010356A3 (fr) * 2005-07-18 2007-08-23 Pfizer Ltd Procede pour la preparation de derives de sulfonamide
CN102051388B (zh) * 2005-07-18 2013-03-27 辉瑞有限公司 化合物的制备方法

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AU3500500A (en) 2000-09-14
AR022725A1 (es) 2002-09-04

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