Direct observation of a fluorinated anticonvulsant in brain tissue using 19F-NMR techniques

Biochem Pharmacol. 1993 Feb 24;45(4):949-59. doi: 10.1016/0006-2952(93)90181-u.

Abstract

A fluorinated derivative of an anticonvulsant gamma-butyrolactone [alpha-(1,1-difluoroethyl)-alpha-methyl-gamma-butyrolactone; alpha-DFGBL] was synthesized as a probe for NMR spectroscopic observation of the drug in brain tissue. The fluorinated compound is an efficacious anticonvulsant in mice, and inhibits the specific binding of [35S]t-butylbicyclophosphorothionate ([35S]TBPS) to mouse brain membranes with a concentration dependence similar to that of the non-fluorinated compound alpha-ethyl-alpha-methyl-gamma-butyrolactone. Quantitative 19F-NMR spectroscopic studies, coupled with chromatographic measurements of drug tissue concentration, showed that virtually all of the alpha-DFGBL in brain was NMR-observable and that, following intraperitoneal injection, alpha-DFGBL rapidly achieved millimolar concentrations in brain. The 19F-NMR spectra of a alpha-DFGBL in brain and liver tissue were broad (1-2 ppm) and complex, exhibiting multiple chemical shift features. The major chemical shift features in these spectra were assigned on the basis of differential extraction and comparison of 19F spin-spin relaxation times (T2s) and 19F chemical shifts of alpha-DFGBL in tissue to those in pure solvents. The major feature at 10.4 ppm in the tissue spectra was assigned to a weakly polar, membrane-associated environment for the fluorinated compound, while the feature at 11.2 ppm was assigned to an aqueous environment for alpha-DFGBL. The drug was in slow exchange between these two environments in brain. In addition, the feature at lowest field (9.7-9.8 ppm) was identified as a water-soluble hydroxy-acid metabolite of alpha-DFGBL produced by the liver. These data indicate that gamma-butyrolactone anticonvulsants achieve high concentrations in brain, where they exist in several, largely membrane-associated, environments. These findings are consistent with the purported action of the gamma-butyrolactones as low-affinity modulators of gamma-aminobutyric acid-A channels.

Publication types

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

MeSH terms

  • 4-Butyrolactone / analogs & derivatives*
  • 4-Butyrolactone / analysis
  • 4-Butyrolactone / chemical synthesis
  • 4-Butyrolactone / pharmacokinetics
  • Adipose Tissue / chemistry
  • Animals
  • Anticonvulsants / analysis*
  • Brain / metabolism
  • Brain Chemistry*
  • Bridged Bicyclo Compounds / metabolism
  • Bridged Bicyclo Compounds, Heterocyclic*
  • Chromatography, Gas
  • Convulsants / metabolism
  • Liver / chemistry
  • Liver / metabolism
  • Magnetic Resonance Spectroscopy / methods
  • Mice
  • Tissue Distribution

Substances

  • Anticonvulsants
  • Bridged Bicyclo Compounds
  • Bridged Bicyclo Compounds, Heterocyclic
  • Convulsants
  • alpha-(1,1,-difluoroethyl)-alpha-methyl-gamma-butyrolactone
  • tert-butylbicyclophosphorothionate
  • 4-Butyrolactone