A new high-performance liquid chromatography method determines low production of 9-beta-D-arabinofuranosylguanine triphosphate, an active metabolite of nelarabine, in adult T-cell leukemia cells.

Abstract

The 9-beta-D-arabinofuranosylguanine (ara-G), an active compound of nelarabine, demonstrates potent cytotoxicity specifically on T-cell malignancies. In cells, ara-G is phosphorylated to ara-G triphosphate (ara-GTP), which is subsequently incorporated into DNA, thereby inhibiting DNA synthesis. Because ara-GTP is crucial to ara-G's cytotoxicity, the determination of ara-GTP production in cancer cells is informative for optimizing nelarabine administration. Here, we developed a new, sensitive isocratic-elution HPLC method for quantifying ara-GTP. Samples were eluted isocratically by using phosphate buffer at a constant flow rate. Ara-GTP was clearly separated from other nucleotides by using an anion-exchange column and it was quantitated by its peak area at 254 nm. The standard curve was linear with low variability and a sensitive detection limit (10 pmol). Furthermore, due to ara-G's specificity to T-cells we hypothesized that nelarabine might be effective against adult T-cell leukemia (ATL). The ara-GTP production was compared between T-lymphoblastic leukemia CCRF-CEM and ATL cell lines in vitro. When CEM cells were incubated with ara-G, the ara-GTP production increased in a concentration- and time-dependent manner. In contrast, 5 ATL cell lines accumulated lower ara-GTP in the same condition. While ara-G inhibited the growth of CEM cells with a 50% growth inhibition concentration of 2 microM, the inhibitory-concentration values were >1 mM in 8 of the 12 ATL cell lines. This ineffectiveness appeared to correspond with the low ara-GTP production. The present study is the first to evaluate the potential of ara-G against ATL cells; our results suggest that nelarabine would not be effective against ATL.

• Authors: Takahiro Yamauchi, Rie Nishi, Kazuhiro Kitazumi, Tsuyoshi Nakano, Takanori Ueda
• Journal: Oncology reports (Oncol Rep) Vol. 23 Issue 2 Pg. 499-504 (Feb 2010) ISSN: 1791-2431 [Electronic] Greece
• PMID: 20043113 (Publication Type: Journal Article, Validation Study)

Chemical References

• Antineoplastic Agents
• Arabinonucleosides
• Arabinonucleotides
• Biomarkers
• Prodrugs
• 9-arabinofuranosylguanine
• nelarabine
• 9-beta-D-arabinofuranosylguanosine 5'-triphosphate
• Guanosine Triphosphate

Topics

• Antineoplastic Agents (metabolism, therapeutic use)
• Arabinonucleosides (metabolism, pharmacokinetics, pharmacology, therapeutic use)
• Arabinonucleotides (analysis, metabolism)
• Biomarkers (analysis, metabolism)
• Cell Proliferation (drug effects)
• Chemical Fractionation (methods)
• Chromatography, High Pressure Liquid (economics, methods, standards)
• Guanosine Triphosphate (analogs & derivatives, analysis, metabolism)
• Humans
• Leukemia-Lymphoma, Adult T-Cell (diagnosis, drug therapy, metabolism)
• Prodrugs (metabolism, pharmacokinetics)
• Reference Standards
• Sensitivity and Specificity
• Treatment Outcome
• Tumor Cells, Cultured