Microbial transformation method for the preparation of an epothilone

• US 6,780,620 B1
• Filed: 12/21/1999
• Issued: 08/24/2004
• Est. Priority Date: 12/23/1998
• Status: Expired due to Term

• Current Assignee: Bristol-Myers Squibb Company
• Original Assignee: Bristol-Myers Squibb Company
• Inventors: Huang, Xiaohua, Lam, Kin Sing, Li, Wenying, Matson, James A., McClure, Grace A.
• Primary Examiner(s): Lilling, Herbert J.
• Application Number:
  US09/468,854
• Time in Patent Office: 1,708 Days
• Field of Search: 435/117, 435/118, 435/119, 435/121, 435/123
• US Class Current: 435/117
• CPC Class Codes: C12P 17/08 containing a hetero ring of at least seven ring members, e.g. zearalenone, macrolide aglycons
• CPC Class Codes: C12P 17/10 Nitrogen as only ring hetero atom
• CPC Class Codes: C12P 17/18 containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin e.g. Rifamycin C12P17/189
• 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: C12P 17/182 Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system alloxazine or isoalloxazine, e.g. riboflavine C12P25/00
• CPC Class Codes: C12P 17/185 Heterocyclic compounds containing sulfur atoms as ring hetero atoms in the condensed system cepam nucleus C12P35/00; penam nucleus C12P37/00

Abstract

A microbial method for the preparation of an epothilone containing a terminal hydroxyalkyl group, comprising contacting at least one epothilone having a terminal alkyl group with an enzyme or microorganism capable of catalyzing the selective hydroxylation of said alkyl group to a hydroxyalkyl group, and effecting said hydroxylation.

Claims

1. A method for the preparation of at least one hydroxyalkyl-bearing compound of the following formula III:

where

Q is selected from the group consisting of

G₁ is the following formula V

2. The method of claim 1 wherein said compound of formula III is epothilone F and said compound of formula IV is epothilone B.

3. The method of claim 1 wherein n is zero and m is 1.

4. The method of claim 1 wherein n is zero, m is 1, and A₂ is alkenyl.

5. The method of claim 1 wherein said microorganism is selected from the group consisting of Actinomycetes, Amycolata, Amycolatopsis, Beauveria, Candida, Gilbertella, Nocardia, Pseudomonas, Saccharopolyspora, Saccharothrix and Streptomyces.

6. The method of claim 1 wherein said microorganism is selected selected from the group consisting of Amycolata autotrophica, Beauveria basiana, Candida rugosa, and Pseudomonas putida.

7. The method of claim 1 wherein said microorganism is selected from the group consisting of Amycolata autotrophica ATCC 35203 and Actinomycetes sp. strain SC 15847 PTA-1043.

8. The method of claim 1, wherein said microorganism is Actinomycetes sp. strain SC 15847 PTA-1043.

9. The method of claim 1 wherein said method comprises a biotransformation under submerged aerobic conditions in an aqueous medium.

10. The method of claim 9, wherein said aqueous medium contains carbon and nitrogen nutrients at approximately pH 5-8.

11. The method of claim 1, wherein said P-450 enzyme is isolated from a microorganism selected from the group consisting of Actinomycetes, Amycolata, Amycolatopsis, Beauveria, Candida, Gilbertella, Nocardia, Pseudomonas, Saccharopolyspora, Saccharothrix and Streptomyces.

12. The method of claim 1 wherein said P-450 enzyme is isolated from a microorganism selected from the group consisting of Amycolata autotrophica ATCC 35203 and Actinomycetes sp. strain SC 15847 PTA-1043.

13. The method of claim 1, wherein G₁ is

14. The method of claim 1, wherein G₂ is

15. The method of claim 1, wherein Q is

16. The method of claim 1, wherein Q is

17. The method of claim 1, wherein the compound of formula (III) is a compound having the formula,

wherein W is O or NH.

18. The method of claim 17, wherein G₁ is

19. The method of claim 14, wherein G₂ is

20. The method of claim 17, wherein Q is

21. The method of claim 17, wherein Q is