Engineering an enantioselective amine oxidase for the synthesis of pharmaceutical building blocks and alkaloid natural products

J Am Chem Soc. 2013 Jul 24;135(29):10863-9. doi: 10.1021/ja4051235. Epub 2013 Jul 15.

Abstract

The development of cost-effective and sustainable catalytic methods for the production of enantiomerically pure chiral amines is a key challenge facing the pharmaceutical and fine chemical industries. This challenge is highlighted by the estimate that 40-45% of drug candidates contain a chiral amine, fueling a demand for broadly applicable synthetic methods that deliver target structures in high yield and enantiomeric excess. Herein we describe the development and application of a "toolbox" of monoamine oxidase variants from Aspergillus niger (MAO-N) which display remarkable substrate scope and tolerance for sterically demanding motifs, including a new variant, which exhibits high activity and enantioselectivity toward substrates containing the aminodiphenylmethane (benzhydrylamine) template. By combining rational structure-guided engineering with high-throughput screening, it has been possible to expand the substrate scope of MAO-N to accommodate amine substrates containing bulky aryl substituents. These engineered MAO-N biocatalysts have been applied in deracemization reactions for the efficient asymmetric synthesis of the generic active pharmaceutical ingredients Solifenacin and Levocetirizine as well as the natural products (R)-coniine, (R)-eleagnine, and (R)-leptaflorine. We also report a novel MAO-N mediated asymmetric oxidative Pictet-Spengler approach to the synthesis of (R)-harmicine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaloids / chemistry
  • Alkaloids / metabolism*
  • Aspergillus niger / chemistry
  • Aspergillus niger / enzymology*
  • Aspergillus niger / genetics
  • Aspergillus niger / metabolism
  • Benzhydryl Compounds / chemistry
  • Benzhydryl Compounds / metabolism
  • Biological Products / chemistry
  • Biological Products / metabolism
  • Indole Alkaloids / chemistry
  • Indole Alkaloids / metabolism
  • Models, Molecular
  • Monoamine Oxidase / genetics*
  • Monoamine Oxidase / metabolism
  • Oxidation-Reduction
  • Protein Engineering / methods*
  • Stereoisomerism

Substances

  • Alkaloids
  • Benzhydryl Compounds
  • Biological Products
  • Indole Alkaloids
  • harmicine
  • benzhydrylamine
  • Monoamine Oxidase