Microbial process for the preparation of milbemycin derivatives

• US 5,183,749 A
• Filed: 03/20/1990
• Issued: 02/02/1993
• Est. Priority Date: 02/04/1987
• Status: Expired due to Fees

• Current Assignee: Novartis Corporation (Novartis Ag)
• Original Assignee: Sankyo Company Limited (Daiichi Sankyo Company Limited)
• Inventors: Frei, Bruno, Ramos, Gerado, Schar, Hans-Peter, Maienfisch, Peter, O'Sullivan, Anthony C., Ghisalba, Oreste
• Primary Examiner(s): Marx, Irene
• Application Number:
  US07/496,183
• Time in Patent Office: 1,050 Days
• Field of Search: 435/119, 435/58, 435/886, 435/917, 435/822, 435/911, 435/254, 435/253.5
• US Class Current: 435/119
• CPC Class Codes: A01N 49/00 Biocides, pest repellants or attractants, or plant growth regulators, containing compounds containing the group , wherein m+n>=1, both X together may also mean —Y— or a direct carbon-to-carbon bond, and the carbon atoms marked with an asterisk are not part of any ring system other than that which may be formed by the atoms X, the carbon atoms in square brackets being part of any acyclic or cyclic structure, or the group , wherein A means a carbon atom or Y, n>=0, and not more than one of these carbon atoms being a member of the same ring system, e.g. juvenile insect hormones or mimics thereof containing hydrocarbons A01N27/00
• CPC Class Codes: C07H 19/01 sharing oxygen
• CPC Class Codes: C12N 1/145 Fungal isolates
• CPC Class Codes: C12N 1/205 Bacterial isolates
• CPC Class Codes: C12P 17/181 Heterocyclic compounds containing oxygen atoms as the only ring heteroatoms in the condensed system, e.g. Salinomycin, Septamycin
• CPC Class Codes: C12R 2001/525 Streptomyces diastatochromogenes
• CPC Class Codes: C12R 2001/685 Aspergillus niger
• CPC Class Codes: Y10S 435/822 using bacteria or actinomycetales
• CPC Class Codes: Y10S 435/886 Streptomyces
• CPC Class Codes: Y10S 435/911 using fungi
• CPC Class Codes: Y10S 435/917 Aspergillus niger

Abstract

The present invention relates to a one-step microbial process for the preparation of 13β-hydroxy- or 14,15-epoxy-milbemycins of the following formulae I and II ##STR1## in which R₁ and R₁ '"'"' represent methyl, ethyl, isopropyl or sec.-butyl or represent the group ##STR2## in which X represents methyl, ethyl or isopropyl, and A and A'"'"' represent the group ##STR3## in which R₂ represents hydrogen, --C(O)-CH₂ O-C(O)-CH₂ CH₂ -COOH or --C(O)-CH₂ CH₂ -COOH, or mixtures of compounds of the formulae I and II, which process comprises bringing a milbemycin of the formula III ##STR4## dissolved in a liquid phase, in which R₃ represents R₁ or R₁ '"'"' and R₄ represents A or A'"'"', and R₁, R₁ '"'"', A and A'"'"' have the meanings given for formulae I and II, into contact with a microorganism that is capable of 13β-hydroxylation or 14,15-epoxidation or of both reactions, or into contact with active constituents thereof. The microorganism or its active constituents may be either in free form or immobilized form.

Claims

1. A one-step microbial process for the preparation of 13β

-hydroxy-milbemycins or 14,15-epoxy-milbemycins of formulas I or II;

##STR51## in which R₁ and R₁ '"'"' are methyl or isopropyl and A and A'"'"' are --CH(OH)--, or R₁ and R₁ '"'"' are ethyl and A and A'"'"' are --CH(OH)-- or --C(═

N˜

OH)--,

or a mixture of compounds of formulas I and II, which process comprises bringing a milbemycin of formula III ##STR52## dissolved in a liquid phase, in which R₃ is R₁ or R₁ '"'"' and R₄ is A or A'"'"', and R₁, R₁ '"'"', A and A'"'"' have the meanings given for formulas I and II, into contact with a microorganism selected from the group consisting of Streptomyces diastatochromogenes ATCC 31561, Streptomyces violascens ATCC 31560 and Aspergillus niger ATCC 20567 that is capable of 13β

-hydroxylation or 14,15-epoxidation, or of both reactions, for a period of time that is sufficient for carrying out the 13β

-hydroxylation or 14,15-epoxidation reaction or both reactions, and isolating the reaction product.

2. A process of claim 1, wherein the microorganism is Streptomyces violascens ATCC 31560.

3. A process of claim 1, wherein the microorganism is Streptomyces diastatochromogenes ATCC 31561.

4. A process of claim 1, wherein the microorganism is Aspergillus niger ATCC 20567.

5. A process of claim 1, wherein the microorganism is in immobilized form.

6. A process of claim 1, wherein the hydroxylation or epoxidation reaction or both reactions are carried out in a bioreactor.

7. A process of claim 6, wherein the bioreactor is a stirred vessel reactor.

8. A process of claim 1, wherein the process temperature is in the range of 20°

C. to 32°

C.

9. A process of claim 1, wherein the milbemycin of formula III is brought into contact with the microorganism in a culture medium at a pH in the range of 4 to 8.

10. A process of claim 1, wherein the milbemycin of formula III is brought into contact with the microorganism in a culture medium having a nitrogen source at a concentration in the range of 0.1 g/l to 6 g/l.

11. A process of claim 1, wherein the milbemycin of formula III is brought into contact with the microorganism in a culture medium having a carbon source at a concentration in the range of 1.0 g/l to 25 g/l.