Chemical Decomposition of 5-Aza-2′-deoxycytidine (Decitabine): Kinetic Analyses and Identification of Products by NMR, HPLC, and Mass Spectrometry

• Published in: Chemical research in toxicology 2009-06, Vol.22 (6), p.1194-1204
• Main Authors: Rogstad, Daniel K, Herring, Jason L, Theruvathu, Jacob A, Burdzy, Artur, Perry, Christopher C, Neidigh, Jonathan W, Sowers, Lawrence C
• Format: Article
• Language: English
• Subjects: Azacitidine - analogs & derivatives; Azacitidine - chemistry; Chromatography, High Pressure Liquid; DNA Modification Methylases - antagonists & inhibitors; DNA Modification Methylases - metabolism; Enzyme Inhibitors - chemistry; Gas Chromatography-Mass Spectrometry; Half-Life; Hydrogen-Ion Concentration; Kinetics; Magnetic Resonance Spectroscopy; Spectrophotometry, Ultraviolet; Temperature; Time Factors
• ISSN: 0893-228X; 1520-5010
• DOI: 10.1021/tx900131u

The nucleoside analogue 5-aza-2′-deoxycytidine (Decitabine, DAC) is one of several drugs in clinical use that inhibit DNA methyltransferases, leading to a decrease of 5-methylcytosine in newly replicated DNA and subsequent transcriptional activation of genes silenced by cytosine methylation. In addition to methyltransferase inhibition, DAC has demonstrated toxicity and potential mutagenicity, and can induce a DNA-repair response. The mechanisms accounting for these events are not well understood. DAC is chemically unstable in aqueous solutions, but there is little consensus between previous reports as to its half-life and corresponding products of decomposition at physiological temperature and pH, potentially confounding studies on its mechanism of action and long-term use in humans. Here, we have employed a battery of analytical methods to estimate kinetic rates and to characterize DAC decomposition products under conditions of physiological temperature and pH. Our results indicate that DAC decomposes into a plethora of products, formed by hydrolytic opening and deformylation of the triazine ring, in addition to anomerization and possibly other changes in the sugar ring structure. We also discuss the advantages and problems associated with each analytical method used. The results reported here will facilitate ongoing studies and clinical trials aimed at understanding the mechanisms of action, toxicity, and possible mutagenicity of DAC and related analogues.