Milacemide metabolism in rat liver and brain slices by solids NMR

JR Garbow, WP Ridley, V Bork, Y Pan and J Schaefer

Monsanto Company, St. Louis, MO 63198.

The metabolism of 13C- and 15N-labeled milacemide, 2-(pentylamino)- acetamide, has been studied in rat liver and brain slices using solid- state NMR. This analysis is fast and efficient and can be used to monitor both major and minor metabolic pathways in mammalian tissue culture. The NMR work reported herein involves both conventional cross- polarization magic-angle spinning 13C and 15N NMR spectra and rotational-echo double resonance 13C-15N experiments. The latter measure quantitatively the breaking of isotopically labeled carbon- nitrogen chemical bonds. Our results, which are consistent with suggestions from previous metabolic studies, show that the first step in the breakdown of milacemide is the breaking of the pentylamine nitrogen bond to yield pentanoic acid and glycinamide. Total incorporation of 15N label from the resulting glycinamide fragment is comparable in rat liver and brain. In both tissues, considerably more of the 15N label from glycinamide is incorporated than the corresponding 13C label. Differences between the liver and brain tissue are also observed, with more synthesis incorporating the 13C labels taking place in the liver.

Volume 22, Issue 2, pp. 298-303, 03/01/1994
Copyright © 1994 by American Society for Pharmacology and Experimental Therapeutics