Substrate/Product-Targeted NMR Monitoring of Pyrimidine Catabolism and Its Inhibition by a Clinical Drug

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• 作者：Hisatsugu Yamada ; Keigo Mizusawa ; Ryuji Igarashi ; Hidehito Tochio ; Masahiro Shirakawa ; Yasuhiko Tabata ; Yu Kimura ; Teruyuki Kondo ; Yasuhiro Aoyama ; Shinsuke Sando
• 刊名：ACS Chemical Biology
• 出版年：2012
• 出版时间：March 16, 2012
• 年：2012
• 卷：7
• 期：3
• 页码：535-542
• 全文大小：373K
• 年卷期：v.7,no.3(March 16, 2012)
• ISSN：1554-8937

文摘

We report the application of one-dimensional triple-resonance NMR to metabolic analysis and thereon-based evaluation of drug activity. Doubly ¹³C/¹⁵N-labeled uracil ([¹⁵N1,¹³C6]-uracil) was prepared. Its catabolic (degradative) conversion to [¹³C3,¹⁵N4]鈭捨?alanine and inhibition thereof by gimeracil, a clinical co-drug used with the antitumor agent 5-fluorouracil, in mouse liver lysates were monitored specifically using one-dimensional triple-resonance (¹H鈥搟¹³C鈥?sup>15N}) NMR, but not double-resonance (¹H鈥搟¹³C}) NMR, in a ratiometric manner. The administration of labeled uracil to a mouse resulted in its non-selective distribution in various organs, with efficient catabolism to labeled 尾-alanine exclusively in the liver. The co-administration of gimeracil inhibited the catabolic conversion of uracil in the liver. In marked contrast to in vitro results, however, gimeracil had practically no effect on the level of uracil in the liver. The potentiality of triple-resonance NMR in the analysis of in vivo pharmaceutical activity of drugs targeting particular metabolic reactions is discussed.