Synthesis, transport and pharmacokinetics of 5'-amino acid ester prodrugs of 1-beta-D-arabinofuranosylcytosine

Mol Pharm. 2009 Jan-Feb;6(1):315-25. doi: 10.1021/mp800200a.


Cytarabine (1-beta-d-arabinofuranosylcytosine, ara-C, 1) suffers from low oral bioavailability due to low intestinal membrane permeability and poor metabolic stability, and intravenous infusion is usually adopted as the clinical standard dosing administration. To develop an oral alternative for 1 and utilize the intestinal oligopeptide transporter 1 (PepT1), a series of 5'-amino acid ester derivatives of 1 was synthesized to clarify which modification was the most suitable to increase the oral bioavailability of 1. Their apical-to-basolateral permeability across Caco-2 cells and the antiproliferative activity with HL-60 cells were screened. 5'-Valyl prodrug 2 demonstrated the highest permeability and was selected for further study. Glycylsarcosine (gly-sar, a typical substrate of PepT1) uptake by Caco-2 cells can be inhibited by 2 in a concentration-dependent manner, and IC(50) for 2 was 2.18 +/- 0.12 mM. The uptake of 2 was markedly increased in the long-term leptin-treated Caco-2 cells compared with the control Caco-2 cells, and was significantly inhibited by the excess of gly-sar, but not by l-valine. A dose-proportional pharmacokinetics was observed in rats when 5, 15, 30 mg/kg doses of 2 (calculated as 1) were orally administered. The oral absolute bioavailability of 1 was 60.0% and 21.8% after 2 and 1 were orally administered to rats 30 mg/kg, respectively. Following oral administration of 15 mg/kg, the absorption and bioactivation of 2 were extensive and rapid, over 98% of prodrug hydrolysis occurring before appearance in the portal vein. The in vivo dispositions of 1-beta-D-arabinofuranosyluracil (ara-U), a deaminated product of 1, were investigated. Oral administration of 2 resulted in an increased 1/ara-U ratio (2.76) in the blood, much higher than that (1.25) after 1 orally taken. Overall, these results demonstrated that the PepT1-mediated absorption of 2 and the increased metabolic stability resulted in a dramatic increase in the oral bioavailability of 1 in rats and further corroborated the thought that prodrug design strategy targeting intestinal PepT1 was an important and promising strategy to improve oral bioavailability of poorly absorbed drugs.

MeSH terms

  • Amino Acids / chemistry*
  • Animals
  • Caco-2 Cells
  • Cell Proliferation / drug effects
  • Cytarabine / analogs & derivatives*
  • Cytarabine / chemical synthesis*
  • Cytarabine / chemistry
  • Cytarabine / metabolism
  • Cytarabine / pharmacology*
  • Esterification
  • Humans
  • Molecular Structure
  • Prodrugs / chemical synthesis*
  • Prodrugs / chemistry
  • Prodrugs / metabolism
  • Prodrugs / pharmacokinetics*
  • Rats
  • Temperature


  • Amino Acids
  • Prodrugs
  • Cytarabine