Analogues of Carbamyl Aspartate as Inhibitors of Dihydroorotase: Preparation of Boronic Acid Transition-State Analogues and a Zinc Chelator Carbamylhomocysteine

J Med Chem. 1990 Feb;33(2):819-23. doi: 10.1021/jm00164a055.

Abstract

Dihydroorotase (DHO) catalyzes the conversion of carbamyl aspartate (CA) to dihydroorotate (DO) in the de novo pyrimidine biosynthetic pathway. Few effective inhibitors of DHO have been reported, and thus blockade of this reaction has not been widely pursued as a strategy for development of antitumor agents. Utilizing two mechanism-based strategies, we have designed and prepared potential DHO inhibitor analogues of CA. One strategy replaced the gamma-carboxyl moiety of CA with a boronic acid. This substitution yields compounds which form stable charged tetrahedral intermediates and mimic the enzyme-substrate transition state. Preparation of the boronic acid analogues of CA and its carboxylic acid esters focused on a Curtius rearrangement as a key step following a malonic ester synthesis. This was followed by carbamoylation of the free amine under nonaqueous neutral conditions with Si(NCO)4. The ethyl ester was a competitive inhibitor of DHO with an apparent Ki of 5.07 microM, while the nonesterified analogue and the methyl ester were not effective inhibitors. None of the compounds were cytotoxic against L1210 cells in culture. An active-site-directed sulfhydryl-containing zinc chelator was also prepared. This analogue irreversibly inhibited the enzyme, but it also was ineffective in L1210 growth inhibition.

Publication types

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

MeSH terms

  • Amidohydrolases / antagonists & inhibitors*
  • Animals
  • Boronic Acids*
  • Cell Division / drug effects
  • Chelating Agents
  • Dihydroorotase / antagonists & inhibitors*
  • Drug Design
  • Enzyme Inhibitors / chemical synthesis*
  • Homocysteine*
  • Leukemia L1210
  • Tumor Cells, Cultured / drug effects
  • Zinc

Substances

  • Boronic Acids
  • Chelating Agents
  • Enzyme Inhibitors
  • Homocysteine
  • Amidohydrolases
  • Dihydroorotase
  • Zinc