Enzymatic oxidative deamination process

Details Enzymatic oxidative deamination process Enzymatic oxidative deamination process

2001-04-26

2003-11-18

Prouty, Rebecca E. (Department: 1652)

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Transferase other than ribonuclease

C435S252300, C435S320100, C435S069100, C435S252330, C435S254100, C536S023200

Reexamination Certificate

active

06649387

ABSTRACT:

FIELD OF INVENTION
The present invention concerns an enzymatic oxidative deamination process to prepare an intermediate useful to prepare compounds having endopeptidase and angiotensin converting enzyme inhibition activity.
BACKGROUND OF THE INVENTION
Robl in U.S. Pat. No. 5,508,272 discloses compounds of the formula
wherein A is
as possessing neutral endopeptidase and angiotensin converting enzyme inhibition activity. Among these compounds is [4S-[4&agr;(R*),7&agr;,10a&bgr;]]-octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylic acid which is currently undergoing clinical evaluation. This compound is reported herein as (1).
Robl discloses that the amino lactam portion of (1), i.e., the intermediate
can be prepared by coupling (S)-2-amino-6,6-dimethoxyhexanoic acid methyl ester with the N-protected amino acid
wherein P
1
is an amino protecting group and P
2
is a sulfur protecting group to give the dipeptide of the formula
Removal of the P
2
protecting group, followed by acid catalyzed cyclization, and removal of the P
1
protecting group gives [4S-(4&agr;,7&agr;,10a&bgr;)]-octahydro-4-amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazephine-7-carboxdylic acid, methyl ester.
Robl discloses preparing (S)-2-amino-6,6-dialkoxyhexanoic acid, alkyl ester, such as (S)-2-amino-6,6-dimethoxyhexanoic acid, methyl ester, by converting N-protected L-&egr;-hydroxynorleucine to its methyl ester, oxidizing to a corresponding aldehyde, such as of the formula
then reacting with trimethyl orthoformate in the presence of a strong acid catalyst, and removing the P
3
protecting group.
SUMMARY OF THE INVENTION
The present invention provides an enzymatic process for the preparation of the above-described intermediate, i.e., the protected amino lactam portion of (1).
More specifically, the present invention is directed to a process for preparing an amino lactam compound of the formula I
comprising contacting a dipeptide monomer of the formula II
wherein P
1
is an amino protecting group, and R
1
is H, alkyl or of the formula
wherein R
2
is alkyl,
with an aminotransferase enzyme in the presence of &agr;-ketoglutarate under conditions suitable for formation of the compound of formula I. The immediately preceding process will be referred to herein as the “oxidative deamination” process.
In an alternate embodiment, the oxidative deamination process is performed in the presence of a glutamate oxidase enzyme which functions to recycle glutamate formed during the process back to alpha-ketoglutarate.
The present invention is also directed to a process for preparing a dipeptide monomer starting compound of the formula III
comprising contacting a dipeptide dimer compound of the formula IV
wherein P
1
is as defined above,
with a reducing agent under conditions suitable for formation of the compound of formula III. The immediately preceding process for preparing the dipeptide monomer will be referred to herein as the “reduction” process.
The present invention also concerns an engineered host cell containing recombinant nucleic acid capable of expressing an aminotransferase enzyme. In addition, the invention concerns a novel aminotransferase from Spingomonas sp. and nucleic acid encoding same.
The present invention also concerns the novel compounds of formulas II, III and IV.


REFERENCES:
patent: 5508272 (1996-04-01), Robl
patent: 6261810 (2001-07-01), Patel et al.
patent: 0 036 776 (1988-11-01), None
Hames et al., Nucleic Acid Hybridization, A Practical Approach, IRL Press, Washington, D.C. (1985), Title and publication pages, and pp. 113-137 (Chapter 5).
Kern, B. et al., Antimicrobial Agents and Chemotherapy, American Society of Microbiology, vol. 17, No. 4, pp. 679-685 (1986).
Madduri, K. et al., Journal of Bacteriology, vol. 173, No. 3, pp. 985-988 (1991).
Coque, J. et al., Journal of Bacteriology, vol. 173, No. 19, pp. 6258-6264 (1991).
Houghton et al., Proc.Natl Acad.Sci. 82, 5131-5135 (1985).
Devereux et al., Nucl.Acids Res. 12:387-395 (1984).
Needleman et al., J. Mol. Biol. 48:443-453 (1970).
Smith et al., Adv.Appl.Math 2:482 (1981).
Gribskov et al., Nucl. Acids Res. 14:6745-6763 (1986).
Schwartz et al., Atlas of Protein Sequence and Structure, National Biomedical Research Foundation, 353-358 (1979).
White et al., Trends Genet. 5:185-189 (1989).
Morinaga et al., Nucl. Acids Res. 2:636-639 (1984).
Taylor et al., Nucl, Acids Res. 13: 8746-8764 (1985).
Kunkel, Proc. Natl. Acad. Sci USA 82:482-492 (1985).
Sayers et al., Nucl. Acids Res. 16:791-802 (1988).
Takada et al., J. Biochem. 109:371-376 (1991).
Bolivar et al., Gene 2:95-113 (1977).
Chang et al., Nature, 275:617-624 (1978).
Goeddel et al., Nature 281:544-548 (1979).
Goeddel et al., Nucl. Acids Res. 8:4057-4074 (1980).
Amann et al., Gene 25, 167-178 (1983).
Siebenlist et al., Cell 20:269-281 (1980).
Sanger et al., Proc.Natl.Acad.Sci.USA 74:5463-5467 (1977).
Maxam et al., Proc.Natl.Acad.Sci.USA 74:560-564 (1977).
Park et al. J. Bacteriol 1997, vol. 179, No. 17, pp. 5300-5308.
Oku et al. Biosci. Biotech.Biochem. 1998, vol. 62, No. 4, pp. 622-627.

Affiliated with

Also associated with

Rating

Enzymatic oxidative deamination process does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Enzymatic oxidative deamination process, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Enzymatic oxidative deamination process will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3182215