Degradation pathways of a peptide boronic acid derivative, 2-Pyz-(CO)-Phe-Leu-B(OH)(2)

J Pharm Sci. 2000 Jun;89(6):758-65. doi: 10.1002/(SICI)1520-6017(200006)89:6<758::AID-JPS7>3.0.CO;2-L.

Abstract

The peptide boronic acid derivative 2-Pyz-(CO)-Phe-Leu-B(OH)(2) is a potent inhibitor of 20S proteasome and a proposed anticancer agent. During preformulation studies, the compound presented erratic stability behavior. Efforts were made to isolate and identify the degradation products, thereby helping to identify possible mechanisms for the degradation. The reaction of 2-Pyz-(CO)-Phe-Leu-B(OH)(2) with hydrogen peroxide not only provided a convenient way to isolate the initial degradation products seen from hydrolysis in aqueous buffers but also showed that the major, initial degradation pathway was probably oxidative in nature. The isolated degradation products were characterized by nuclear magnetic resonance spectroscopy, mass spectrometry, and optical rotation dispersion. In the presence of hydrogen peroxide, the boronic acid group was cleaved from 2-Pyz-(CO)-Phe-Leu-B(OH)(2) to give an alcohol with an apparent retention of the original stereochemistry. Subsequent isomerization and further hydrolysis were then seen. Surprisingly, added ascorbate and EDTA accelerated rather than inhibited degradation. Degradation of 2-Pyz-(CO)-Phe-Leu-B(OH)(2) under acidic and basic conditions seemed to be mediated by an initial oxidative degradation pathway similar to that seen with the peroxide.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Ascorbic Acid / chemistry
  • Boronic Acids / chemistry*
  • Chromatography, High Pressure Liquid
  • Hydrogen Peroxide / chemistry
  • Hydrolysis
  • Kinetics

Substances

  • Boronic Acids
  • Hydrogen Peroxide
  • Ascorbic Acid