Biocatalytic asymmetric synthesis of chiral amines from ketones applied to sitagliptin manufacture.
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.
Savile
Christopher K
CK
Codexis, Incorporated, 200 Penobscot Drive, Redwood City, CA 94063, USA. christopher.savile@codexis.com
Janey
Jacob M
JM
Mundorff
Emily C
EC
Moore
Jeffrey C
JC
Tam
Sarena
S
Jarvis
William R
WR
Colbeck
Jeffrey C
JC
Krebber
Anke
A
Fleitz
Fred J
FJ
Brands
Jos
J
Devine
Paul N
PN
Huisman
Gjalt W
GW
Hughes
Gregory J
GJ
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
D000588:Amines; D007004:Hypoglycemic Agents; D007659:Ketones; D011719:Pyrazines; D014230:Triazoles; D000637:Transaminases; D000068900:Sitagliptin Phosphate