In vivo quantification of active decitabine-triphosphate metabolite: a novel pharmacoanalytical endpoint for optimization of hypomethylating therapy in acute myeloid leukemia.
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| Abstract |
Decitabine (DAC) is used for treatment of patients with myelodysplastic syndromes and acute myeloid leukemia (AML). Following cellular uptake, DAC is activated to DAC-triphosphate (TP) and incorporated into DNA. Once incorporated into the DNA, DAC-TP binds and inactivates DNA methyltransferases (DNMTs), thereby leading to hypomethylation and re-expression of epigenetically silenced tumor suppressor genes and ultimately antileukemia activity. However, direct evidence of in vivo DAC-TP occurrence in DAC-treated patients has been difficult to demonstrate due to a lack of suitable validated analytical methodology. Thus, we developed and validated a nonradioactive sensitive and specific LC-MS/MS assay for quantification of DAC-TP. The assay is linear from 50 to 1,000 nM and from 1 to 10 μM and has a lower limit of quantitation of 50 nM and a coefficient of variation for both within- and between-day precision <20%. Following DAC treatment, we detected DAC-TP in parental and DAC-resistant AML cells (in vitro) and bone marrow (BM) and spleen of normal and leukemic mice (in vivo). Downregulation of DNMTs and correlation of DAC-TP concentration with proteins involved in mechanisms of DAC resistance were also demonstrated. The clinical applicability of this method was proven by measuring DAC-TP level in BM and blood mononuclear cells from DAC-treated AML patients. Higher levels are seemingly associated with clinical response. Monitoring the DAC-TP intracellular level may serve as a novel pharmacological endpoint for designing more effective DAC-based regimens.
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| Authors | Hongyan Wang, Ping Chen, Jiang Wang, Ramasamy Santhanam, Josephine Aimiuwu, U V Vijaya Saradhi, Zhongfa Liu, Sebastian Schwind, Alice Mims, John C Byrd, Michael R Grever, Miguel A Villalona-Calero, Rebecca Klisovic, Alison Walker, Ramiro Garzon, William Blum, Kenneth K Chan, Guido Marcucci |
| Journal | The AAPS journal (AAPS J) Vol. 15 Issue 1 Pg. 242-9 (Jan 2013) ISSN: 1550-7416 [Electronic] United States |
| PMID | 23180159 (Publication Type: Journal Article, Research Support, N.I.H., Extramural) |
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