title: Enzymatic synthesis of chiral intermediates for Omapatrilat, an antihypertensive drug.
abstract: Biocatalytic processes were used to prepare chiral intermediates required for the synthesis of Omapatrilat 1 by three different routes. The synthesis and enzymatic conversion of 2-keto-6-hydroxyhexanoic acid 3 to L-6-hydroxynorleucine 2 was demonstrated by reductive amination using beef liver glutamate dehydrogenase. To avoid the lengthy chemical synthesis of the ketoacid 3, a second route was developed to prepare the ketoacid by treatment of racemic 6-hydroxy norleucine [readily available from hydrolysis of 5-(4-hydroxybutyl) hydantoin 4] with D-amino acid oxidase from porcine kidney or Trigonopsis variabilis followed by reductive amination to convert the mixture completely to L-6-hydroxynorleucine in 98% yield and 99% enantiomeric excess (e.e.). The enzymatic synthesis of (S)-2-amino-5-(1,3-dioxolan-2-yl)-pentanoic acid (allysine ethylene acetal, 5) was demonstrated using phenylalanine dehydrogenase (PDH) from T. intermedius. Phenylalanine dehydrogenase was cloned and overexpressed in Escherichia coli and Pichia pastoris. Using PDH from E. coli or P. pastoris, the enzymatic process was scale-up to prepare kg quantity of allysine ethylene acetal 5. The reaction yields of >94% and e.e. of >98% were obtained for allysine ethylene acetal 5. An enzymatic process was developed for the synthesis of [4S-(4a,7a,10ab)]1-octahydro-5-oxo-4 [[(phenylmethoxy)carbonyl]amino]-7H-pyrido-[2,1-b] [1,3]thiazepine-7-carboxylic acid [BMS-199541-01]. The enzymatic oxidation of the epsilon-amino group of lysine in the dipeptide dimer N(2)-[N[[(phenyl-methoxy)carbonyl] L-homocysteinyl] L-lysine)-1,1-disulphide [BMS-201391-01] to produce BMS-199541-01 using a novel L-lysine epsilon-aminotransferase (LAT) from Sphingomonas paucimobilis SC 16113 was demonstrated. This enzyme was overexpressed in E. coli and a process was developed using the recombinant enzyme.
authors.affiliation: Enzyme Technology, Process Research & Development, Bristol-Myers Squibb Pharmaceutical Research Institute, One Squibb Drive, P.O. Box 191, New Brunswick, NJ 08903, USA. patelr@bms.com
authors.forename: R N
authors.initials: RN
authors.lastname: Patel
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000737:chemistry / Q000378:metabolism; D000959:Antihypertensive Agents / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D019149:Bioreactors; D003605:D-Amino-Acid Oxidase / Q000737:chemistry / Q000378:metabolism; D004798:Enzymes / Q000737:chemistry / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D005969:Glutamate Dehydrogenase / Q000737:chemistry / Q000378:metabolism; D051306:L-Lysine 6-Transaminase; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D011725:Pyridines / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013237:Stereoisomerism; D013841:Thiazepines / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D000637:Transaminases / Q000737:chemistry / Q000378:metabolism
chemical_list: D000959:Antihypertensive Agents; D004798:Enzymes; D011725:Pyridines; D011994:Recombinant Proteins; D013841:Thiazepines; C106266:omapatrilat; D000594:Amino Acid Oxidoreductases; D005969:Glutamate Dehydrogenase; C022342:phenylalanine oxidase; D003605:D-Amino-Acid Oxidase; D000637:Transaminases; D051306:L-Lysine 6-Transaminase