Mechanism of Degradation of an α-Keto-Epoxide, a Model for the Warhead for Various Proteasome Inhibitor Anticancer Agents

J Pharm Sci. 2017 Apr;106(4):1051-1061. doi: 10.1016/j.xphs.2016.12.006. Epub 2016 Dec 20.

Abstract

The anticancer agent, carfilzomib, has a unique α-keto-epoxide warhead. The model α-keto-epoxide, N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)pivalamide (1), along with a few of its degradation products was synthesized and studied. The kinetics of hydrolysis and identification of some of the degradation products of 1 were performed at pH values 2, 4, 5, 7, and 8 at 25°C, 40°C, and 60°C and followed by HPLC and liquid chromatography-mass spectroscopy, respectively. 1 degraded independent of pH between pH values 4-7 but showed some acid catalysis at pH 2 and base catalysis at pH 8. Energy of activation, Ea, values progressed from 16.8 ± 0.1 at pH 2 to 20.3 ± 0.1 kcal/mole at pH 8. The major initial degradation products in the pH range 4-5 were the S,R diol (hydrolysis of the epoxide), and S,R chlorohydrin (in the presence of chloride ions). At pH 7-8, the major products were the R,R diastereomer and the S,R and R,R diols. At pH 2, additional unidentified products were seen with relative retention times of 0.28, 0.30, 0.33, and 0.35 and masses equivalent to the diols. The study of 1 provides insight into the degradation of future drugs that use an α-keto-epoxide functional group.

Keywords: cancer chemotherapy; chemical stability; degradation products; hydrolysis; kinetics; stability; stabilization.

MeSH terms

  • Antineoplastic Agents / chemistry*
  • Antineoplastic Agents / metabolism
  • Epoxy Compounds / chemistry*
  • Epoxy Compounds / metabolism
  • Models, Chemical*
  • Pharmaceutical Solutions / chemistry
  • Pharmaceutical Solutions / metabolism
  • Proteasome Inhibitors / chemistry*
  • Proteasome Inhibitors / metabolism

Substances

  • Antineoplastic Agents
  • Epoxy Compounds
  • Pharmaceutical Solutions
  • Proteasome Inhibitors