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Table 3 HPLC analysis of troxacitabine metabolites after exposure to gemcitabine.

From: Synergistic activity of troxacitabine (Troxatyl™) and gemcitabine in pancreatic cancer

Treatment Troxacitabine incorporation
into DNA
(CPM/106 cells)
Metabolites
(pmol/mg protein)
   MP DP TP
[3H]Troxacitabine 10 μM
[3H]Troxacitabine 10 μM + Gemcitabine 10 μM
1248 ± 153
1226 ± 91
5.0 ± 1.6
2.8 ± 0.9
26.0 ± 7.0
13.0 ± 4.0
6.4 ± 1.5
3.2 ± 1.1
[3H]Troxacitabine 10 μM
[3H]Troxacitabine 10 μM + Gemcitabine 0.1 μM
1248 ± 153
1097 ± 298
5.6 ± 1.5
1.7 ± 2.0
22.0 ± 5.0
23.0 ± 2.0
4.5 ± 1.7
5.2 ± 1.0
[3H]Troxacitabine 300 nM
[3H]Troxacitabine 300 nM + Gemcitabine 0.5 nM
4039 ± 118
3838 ± 509
3.0 ± 0.1
2.0 ± 0.2
6.0 ± 0.2
5.9 ± 0.2
2.6 ± 0.3
2.5 ± 0.2
  1. AsPc-1 cells were exposed to [3H]troxacitabine in absence or presence of gemcitabine as described in Materials and Methods. Cells were harvested by trypsinization, washed in cold PBS and extracted with 10% TCA for 10 min on ice. The acid-insoluble pellets, containing DNA into which [3H]troxacitabine was incorporated, were solubilized in 0.5 N NaOH and their radioactive content was determined. Values are presented as (CPM/106 cells ± SD). The supernatants, containing phosphorylated metabolites of [3H]troxacitabine, were analyzed by HPLC. Values (pmol/mg protein ± SEM) for troxacitabine monophosphate (MP), diphosphate (DP) and triphosphate (TP) are given below.