Preparation of an Amino Acid Intermediate for the Dipeptidyl Peptidase IV Inhibitor, Saxagliptin, using a Modified Phenylalanine Dehydrogenase
The non‐proteinogenic amino acid 2‐(3‐hydroxy‐1‐adamantyl)‐(2S)‐aminoethanoic acid [2, (S)‐3‐hydroxyadamantylglycine], is a key intermediate required for the synthesis of Saxagliptin, a dipeptidyl peptidase IV inhibitor under development for treatment of type 2 diabetes mellitus. Keto acid 2‐(3‐hydr...
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Published in | Advanced synthesis & catalysis Vol. 349; no. 8-9; pp. 1369 - 1378 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Weinheim
WILEY-VCH Verlag
04.06.2007
WILEY‐VCH Verlag Wiley |
Subjects | |
Online Access | Get full text |
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Summary: | The non‐proteinogenic amino acid 2‐(3‐hydroxy‐1‐adamantyl)‐(2S)‐aminoethanoic acid [2, (S)‐3‐hydroxyadamantylglycine], is a key intermediate required for the synthesis of Saxagliptin, a dipeptidyl peptidase IV inhibitor under development for treatment of type 2 diabetes mellitus. Keto acid 2‐(3‐hydroxy‐1‐adamantyl)‐2‐oxoethanoic acid (1) was converted to (S)‐3‐hydroxyadamantylglycine by reductive amination using a phenylalanine dehydrogenase from Thermoactinomyces intermedius expressed in a modified form in Pichia pastoris or Escherichia coli. NAD (nicotinamide adenine dinucleotide) produced during the reaction was recycled to NADH (reduced form of nicotinamide adenine dinucleotide) using formate dehydrogenase. Pichia pastoris produces an endogenous formate dehydrogenase when grown on methanol, and the corresponding gene was cloned and expressed in E. coli. The modified phenylalanine dehydrogenase contains two amino acid changes at the C‐terminus and a 12‐amino acid extension of the C‐terminus. The modified enzyme is more effective with keto acid 1 than the wild‐type enzyme, but less effective with the natural substrate, phenylpyruvate. Production of multi‐kg batches was originally carried out with extracts of Pichia pastoris expressing the modified phenylalanine dehydrogenase from Thermoactinomyces intermedius and endogenous formate dehydrogenase, and further scaled up using a preparation of the two enzymes expressed in E. coli. |
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Bibliography: | ark:/67375/WNG-P2TZT0HJ-4 istex:3FC6A35F8FD0D4E2AB7B93EA1ED59D02B53D4FB3 ArticleID:ADSC200700013 ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 |
ISSN: | 1615-4150 1615-4169 |
DOI: | 10.1002/adsc.200700013 |