WO1996023897A1 - Procede de preparation d'une beta-lactamine - Google Patents

Procede de preparation d'une beta-lactamine Download PDF

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Publication number
WO1996023897A1
WO1996023897A1 PCT/NL1996/000052 NL9600052W WO9623897A1 WO 1996023897 A1 WO1996023897 A1 WO 1996023897A1 NL 9600052 W NL9600052 W NL 9600052W WO 9623897 A1 WO9623897 A1 WO 9623897A1
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WO
WIPO (PCT)
Prior art keywords
enzyme
lactam
acylation
process according
core
Prior art date
Application number
PCT/NL1996/000052
Other languages
English (en)
Inventor
Wilhelmus Hubertus Joseph Boesten
Theodorus Johannes Godfried Maria Van Dooren
Johanna Christina Maria Smeets
Original Assignee
Chemferm V.O.F.
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 Chemferm V.O.F. filed Critical Chemferm V.O.F.
Priority to AU48462/96A priority Critical patent/AU4846296A/en
Priority to EP96904337A priority patent/EP0815256A1/fr
Publication of WO1996023897A1 publication Critical patent/WO1996023897A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D501/00—Heterocyclic compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
    • 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
    • C12P35/00—Preparation of compounds having a 5-thia-1-azabicyclo [4.2.0] octane ring system, e.g. cephalosporin

Definitions

  • the invention relates to a process for the preparation of a ⁇ -lactam antibiotic by enzymatic acylation of a ⁇ -lactam core by means of an acylation agent.
  • the enzyme to be applied should be suitable to be re-used many times (for instance more than 50 times, in particular more than 100 times) without much loss of activity.
  • the object of the invention is to obtain a process for the enzymatic acylation of a ⁇ -lactam core whereby said drawback is eliminated.
  • the enzyme activity in successive cycles depends on the concentration of the inorganic salts in the reaction mixture during the enzymatic acylation reaction, as well as on the sum of the concentrations of the ⁇ -lactam antibiotic and the ⁇ -lactam core.
  • ⁇ -lactam cores that can be used in the process according to the invention are penicillic acid derivatives, for instance 6- aminopenicillic acid (6-APA), and cephalosporanic acid derivatives, for instance 7-aminocephalosporanic acid (7- ACA), 7-aminodesacetoxy-cephalosporanic acid (7-ADCA) and 7-amino-3-chlorocephalosporanic (7-ACCA) .
  • penicillic acid derivatives for instance 6- aminopenicillic acid (6-APA)
  • cephalosporanic acid derivatives for instance 7-aminocephalosporanic acid (7- ACA), 7-aminodesacetoxy-cephalosporanic acid (7-ADCA) and 7-amino-3-chlorocephalosporanic (7-ACCA) .
  • Suitable acylation agents that can be used in the process according to the invention are for instance ⁇ - amino acid derivatives, in particular amides and esters of phenyl glycine, p-hydroxyphenyl glycine and dihydrophenyl glycine.
  • any enzyme can be used that is suitable as catalyst in the coupling reaction.
  • Such enzymes are for instance the enzymes that are known under the general designations 'penicillin amidase' and 'penicillin acylase'.
  • suitable enzymes are enzymes derived from Acetobacter, Aeromonas, Alcali ⁇ enes, Aphanocladium, Bacillus SD. , Cephalosporium. Escherichia. Flavobacterium, Kluvvera, Mvcoplana, Protaminobacter, Pseudomonas and Xanthomonas, in particular Acetobacter pasteurianum, Alcali ⁇ enes faecalis, Bacillus me ⁇ aterium.
  • Escherichia coli and Xanthomonas citrii Preferably an immobilized enzyme is used, since the enzyme can be easily re-used then. Immobilized enzymes are known as such and are commercially available. Highly suitable enzymes have appeared to be the Escherichia coli enzyme from Boehringer Mannheim GmbH, which is commercially available under the name 'Enzygel*', the immobilized Penicillin-G acylase from Recordati, the immobilized Penicilline-G acylase from Pharma Biotechnology Hannover, and an Escheria coli penicilline acylase isolated as described in WO-A-92/12782 and immobilised as described in EP-A-222462.
  • the enzymatic acylation reaction is mostly carried out at a temperature between -5 and 35°C, in particular between 5 and 35°C preferably between 0 and 28°C, most preferably between 15 and 28°C.
  • the pH at which the acylation reaction is carried out is mostly between 6 and 8.5.
  • the optimum pH depends on, among other things, the antibiotic, since the stability and the solubility of the ⁇ -lactam antibiotic as well as the ⁇ -lactam core depend on the pH.
  • the pH is preferably between 6.2 and 8.5, most preferably between 7 and 8; if a p-hydroxyphenyl glycine derivative is used as acylation agent the pH is preferably between 6 and 7.5, most preferably between 6 and 7.
  • the enzyme activity is also pH-related.
  • the acylation reaction is mostly carried out in water.
  • the reaction mixture may also contain an organic solvent or a mixture of organic solvents, preferably less than 30 vol.%.
  • organic solvents that can be used are alcohols with 1-7 carbon atoms, for instance a monoalcohol, in particular methanol or ethanol; a diol, in particular ethylene glycol or a triol, in particular glycerol.
  • concentration of the ⁇ -lactam antibiotic and the concentration of the ⁇ -lactam core are chosen such that the sum of the two concentrations is between 200 and 800 mM.
  • concentrations are chosen such that the sum of the two concentrations is between 300 and 700, most preferably between 300 and 600 mM.
  • the molar ratio between the acylating agent and the ⁇ -lactam core is between 0.5:1 and 2:1, preferably between 0.7:1 and 1.3:1, most preferably between 0.8:1 and 1.2:1.
  • the molar ratio between the acylating agent and the ⁇ -lactam core is preferably chosen such that the pH of the reaction mixture throughout the process does not exceed the above-mentioned limits, without titration with acid.
  • inorganic salts will accumulate in the reaction mixture in the presence of anions, for instance Cl", S0 4 2 ", P0 4 3 ", N0 3 ". It has been found that if the concentration of inorganic salts becomes too high, for instance higher than 1000:n mM, where n is the valency of the anion, the enzyme activity declines strongly after several cycles.
  • concentration of inorganic salts in the process according to the invention is therefore kept below 1000:n mM, preferably below 700:n mM, with n as defined above, which implies that for chlorides and nitrates n is 1 and for sulphates it is 2. In order to keep the concentration of inorganic salts low, it is preferred to ensure that the amount of inorganic salts formed during the enzymatic acylation reaction is restricted to a minimum.
  • acylation agents are phenyl glycine and p-hydroxyphenyl glycine derivatives, for instance their amides or esters.
  • these are often used in the form of the salt of a strong acid, for instance phenyl glycine amide.HC1, phenyl glycine amide.*$H 2 S0 4 , phenyl glycine methyl ester.HC1 or dihydrophenyl glycine methyl ester.HC1.
  • the free amide or the free ester is preferably used in order to prevent the formation of additional inorganic salts as a result of neutralization of the salts of the acylation agents.
  • Free amides in solid form are particularly suitable for this purpose.
  • D-PGA D-phenyl glycine
  • Enzyme recycling experiment with 600 mM of D-PGA and 400 mM of 7-ADCA, pH 7.5 - ⁇ 7.8, and titration with 12N H 2 S0 4 .
  • Carried out analogously to example I with 20 g of wet enzyme and a feed consisting of 45.0 g of free D- PGA, 43.7 g of 7-ADCA, 1.8 g of D-PG, 413 ml of water and 5 ml of 25 mass% aqueous NH 4 0H.
  • Initial pH - 7.6, final pH 7.8, kept constant by means of titration with 12N H 2 S0 4 . Reaction time: 2 hours.
  • Enzyme recycling experiment with 600 mM of D-PGA.> $ H 2 S0 4 and 400 mM of 7-ADCA, pH 7.2 - ⁇ 7.5, in the presence of 100 mM of (NH 4 ) 2 S0 4 .
  • Enzyme recycling experiment with 500 mM of D-PGA and 500 mM of 7-ADCA, pH 7.3 ⁇ 7.4, in the presence of 1000 mM of NH 4 C1.
  • Example IX Enzyme recycling experiment with 400 mM of D-PGA and 400 mM Of 7-ADCA, pH 7.3 ⁇ * 7.8.

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  • 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)

Abstract

Procédé de préparation d'une bêta-lactamine par acylation enzymatique d'un noyau bêta-lactame au moyen d'un agent d'acylation, le rapport molaire entre le noyau bêta-lactame et l'agent d'acylation étant compris entre 0,5:1 et 2:1, la concentration de sels minéraux dans le mélange réactionnel étant inférieure à 1000:n mM, où n représente la valence de l'anion, et la somme des concentrations de la bêta-lactamine et du noyau bêta-lactame étant comprise entre 200 et 800 mM. Ce procédé est avantageux en ce que l'enzyme peut être réutilisée à plusieurs reprises sans perte importante de son activité. Par conséquent, on peut obtenir un procédé intéressant sur le plan commercial pour la préparation de bêta-lactamines par acylation enzymatique de noyaux bêta-lactames.
PCT/NL1996/000052 1995-02-02 1996-02-01 Procede de preparation d'une beta-lactamine WO1996023897A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU48462/96A AU4846296A (en) 1995-02-02 1996-02-01 Process for the preparation of a beta -lactam antibiotic
EP96904337A EP0815256A1 (fr) 1995-02-02 1996-02-01 Procede de preparation d'une beta-lactamine

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
BE9500081A BE1009071A3 (nl) 1995-02-02 1995-02-02 Werkwijze voor de bereiding van een beta-lactam antibioticum.
BE9500081 1995-02-02

Publications (1)

Publication Number Publication Date
WO1996023897A1 true WO1996023897A1 (fr) 1996-08-08

Family

ID=3888755

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/NL1996/000052 WO1996023897A1 (fr) 1995-02-02 1996-02-01 Procede de preparation d'une beta-lactamine

Country Status (5)

Country Link
EP (1) EP0815256A1 (fr)
AU (1) AU4846296A (fr)
BE (1) BE1009071A3 (fr)
IN (2) IN181940B (fr)
WO (1) WO1996023897A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999020786A1 (fr) * 1997-10-17 1999-04-29 Dsm N.V. PROCEDE DE PREPARATION D'UN ANTIBIOTIQUE DE β-LACTAME
US6218138B1 (en) * 1998-05-26 2001-04-17 Unifar Kimya Sanayi Ve Ticaret A.S. Synthesis of beta-lactam antibiotics with immobilized penicillin amidase

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991009136A1 (fr) * 1989-12-12 1991-06-27 Consejo Superior Investigaciones Cientificas Procede de synthese d'antibiotiques semi-synthetiques dans des systemes regules thermodynamiquement eau/cosolvants organiques miscibles apolaires par utilisation de penicilline g acylase
WO1992001061A1 (fr) * 1990-07-04 1992-01-23 Novo Nordisk A/S PROCEDE DE PREPARATION DE DERIVES DE β-LACTAMES

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991009136A1 (fr) * 1989-12-12 1991-06-27 Consejo Superior Investigaciones Cientificas Procede de synthese d'antibiotiques semi-synthetiques dans des systemes regules thermodynamiquement eau/cosolvants organiques miscibles apolaires par utilisation de penicilline g acylase
WO1992001061A1 (fr) * 1990-07-04 1992-01-23 Novo Nordisk A/S PROCEDE DE PREPARATION DE DERIVES DE β-LACTAMES

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999020786A1 (fr) * 1997-10-17 1999-04-29 Dsm N.V. PROCEDE DE PREPARATION D'UN ANTIBIOTIQUE DE β-LACTAME
US6287799B1 (en) 1997-10-17 2001-09-11 Dsm N.V. Process for the preparation of aβ-lactam antibiotic
US6218138B1 (en) * 1998-05-26 2001-04-17 Unifar Kimya Sanayi Ve Ticaret A.S. Synthesis of beta-lactam antibiotics with immobilized penicillin amidase

Also Published As

Publication number Publication date
EP0815256A1 (fr) 1998-01-07
AU4846296A (en) 1996-08-21
BE1009071A3 (nl) 1996-11-05
IN181940B (fr) 1998-11-14
IN182469B (fr) 1999-04-17

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