title: Biocatalytic asymmetric synthesis of chiral amines from ketones applied to sitagliptin manufacture.
abstract: Pharmaceutical synthesis can benefit greatly from the selectivity gains associated with enzymatic catalysis. Here, we report an efficient biocatalytic process to replace a recently implemented rhodium-catalyzed asymmetric enamine hydrogenation for the large-scale manufacture of the antidiabetic compound sitagliptin. Starting from an enzyme that had the catalytic machinery to perform the desired chemistry but lacked any activity toward the prositagliptin ketone, we applied a substrate walking, modeling, and mutation approach to create a transaminase with marginal activity for the synthesis of the chiral amine; this variant was then further engineered via directed evolution for practical application in a manufacturing setting. The resultant biocatalysts showed broad applicability toward the synthesis of chiral amines that previously were accessible only via resolution. This work underscores the maturation of biocatalysis to enable efficient, economical, and environmentally benign processes for the manufacture of pharmaceuticals.
authors.affiliation: Codexis, Incorporated, 200 Penobscot Drive, Redwood City, CA 94063, USA. christopher.savile@codexis.com
authors.forename: Christopher K
authors.initials: CK
authors.lastname: Savile
authors.forename: Jacob M
authors.initials: JM
authors.lastname: Janey
authors.forename: Emily C
authors.initials: EC
authors.lastname: Mundorff
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Moore
authors.forename: Sarena
authors.initials: S
authors.lastname: Tam
authors.forename: William R
authors.initials: WR
authors.lastname: Jarvis
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Colbeck
authors.forename: Anke
authors.initials: A
authors.lastname: Krebber
authors.forename: Fred J
authors.initials: FJ
authors.lastname: Fleitz
authors.forename: Jos
authors.initials: J
authors.lastname: Brands
authors.forename: Paul N
authors.initials: PN
authors.lastname: Devine
authors.forename: Gjalt W
authors.initials: GW
authors.lastname: Huisman
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Hughes
mesh_terms: D000588:Amines / Q000138:chemical synthesis*; D055162:Biocatalysis; D020134:Catalytic Domain; D019020:Directed Molecular Evolution; D007004:Hypoglycemic Agents / Q000138:chemical synthesis* / Q000378:metabolism; D007659:Ketones / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D015394:Molecular Structure; D016296:Mutagenesis; D011487:Protein Conformation; D015202:Protein Engineering; D011719:Pyrazines / Q000138:chemical synthesis* / Q000378:metabolism; D000068900:Sitagliptin Phosphate; D012995:Solubility; D013237:Stereoisomerism; D013379:Substrate Specificity; D000637:Transaminases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014230:Triazoles / Q000138:chemical synthesis* / Q000378:metabolism
chemical_list: D000588:Amines; D007004:Hypoglycemic Agents; D007659:Ketones; D011719:Pyrazines; D014230:Triazoles; D000637:Transaminases; D000068900:Sitagliptin Phosphate