Preparative enantiomeric separation of potent AMP-activated protein kinase activator by HPLC on amylose-based chiral stationary phase. Determination of enantiomeric purity and assignment of absolute configuration

J Pharm Biomed Anal. 2008 Apr 14;46(5):920-8. doi: 10.1016/j.jpba.2008.01.048. Epub 2008 Feb 12.

Abstract

The development of high performance liquid chromatography method on amylose-based stationary phases (Chiralpak AD) has permitted to achieve the preparative enantioseparation of one benzimidazole derivative, potent-AMP-kinase (AMPK) activator with satisfactory yields. Analytical enantioseparation method was optimized and validated to determine the enantiomeric purity. Using the UV detection, repeatability, limits of detection (LD) and quantification (LQ) were determined. Single-crystal X-ray analysis was successful to determine the absolute configuration of the individual enantiomers. A relation between the retention order and the absolute configuration of the enantiomers was established.

MeSH terms

  • AMP-Activated Protein Kinases
  • Amylose / analogs & derivatives*
  • Amylose / chemistry
  • Buffers
  • Chromatography, High Pressure Liquid / methods*
  • Crystallography, X-Ray
  • Models, Molecular
  • Molecular Structure
  • Multienzyme Complexes / antagonists & inhibitors*
  • Phenylcarbamates / chemistry*
  • Protein Kinase Inhibitors / chemistry
  • Protein Kinase Inhibitors / isolation & purification*
  • Protein Kinase Inhibitors / pharmacology
  • Protein Serine-Threonine Kinases / antagonists & inhibitors*
  • Reproducibility of Results
  • Solvents / chemistry
  • Spectrophotometry, Ultraviolet
  • Stereoisomerism

Substances

  • Buffers
  • Multienzyme Complexes
  • Phenylcarbamates
  • Protein Kinase Inhibitors
  • Solvents
  • Chiralpak AD
  • Amylose
  • Protein Serine-Threonine Kinases
  • AMP-Activated Protein Kinases