The membrane transport and polyglutamation of pralatrexate: A new-generation dihydrofolate reductase inhibitor

Michele Visentin, Ersin Selcuk Unal, Rongbao Zhao, I. David Goldman

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Purpose: To characterize, directly and for the first time, the membrane transport and metabolism of pralatrexate, a new-generation dihydrofolate reductase inhibitor approved for the treatment for peripheral T-cell lymphoma. Experimental design: [3H]pralatrexate transport was studied in unique HeLa cell lines that express either the reduced folate carrier (RFC) or the proton-coupled folate transporter (PCFT). Metabolism to active polyglutamate derivatives was assessed by liquid chromatography. These properties were compared to those of methotrexate (MTX). Results: The pralatrexate influx K t, mediated by RFC, the major route of folate/antifolate transport at systemic pH, was 0.52 μΜ, 1/10th the MTX influx K i. The electrochemical potential of pralatrexate within HeLa cells far exceeded the extracellular level and was greater than for MTX. In contrast, MTX transport mediated by PCFT, the mechanism of folate/antifolate absorption in the small intestine, exceeded that for pralatrexate. After a 6 h exposure of HeLa cells to 0.5 μM pralatrexate, 80 % of intracellular drug was its active polyglutamate forms, predominantly the tetraglutamate, and was suppressed when cells were loaded with natural folates. There was negligible formation of MTX polyglutamates. The difference in pralatrexate and MTX growth inhibition was far greater after transient exposures (375-fold) than continuous exposure (25-fold) to the drugs. Conclusions: Pralatrexate's enhanced activity relative to MTX is due to its much more rapid rate of transport and polyglutamation, the former less important when the carrier is saturated. The low affinity of pralatrexate for PCFT predicts a lower level of enterohepatic circulation and increased fecal excretion of the drug relative to MTX.

Original languageEnglish (US)
Pages (from-to)597-606
Number of pages10
JournalCancer Chemotherapy and Pharmacology
Volume72
Issue number3
DOIs
StatePublished - Sep 2013

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Folic Acid Antagonists
Methotrexate
Membranes
Proton-Coupled Folate Transporter
Reduced Folate Carrier Protein
HeLa Cells
Folic Acid
Polyglutamic Acid
Metabolism
Pharmaceutical Preparations
Enterohepatic Circulation
Peripheral T-Cell Lymphoma
10-propargyl-10-deazaaminopterin
T-cells
Liquid chromatography
Liquid Chromatography
Design of experiments
Small Intestine
Research Design
Cells

Keywords

  • Methotrexate
  • PCFT
  • Polyglutamation
  • Pralatrexate
  • RFC

ASJC Scopus subject areas

  • Cancer Research
  • Oncology
  • Pharmacology
  • Pharmacology (medical)
  • Toxicology

Cite this

The membrane transport and polyglutamation of pralatrexate : A new-generation dihydrofolate reductase inhibitor. / Visentin, Michele; Unal, Ersin Selcuk; Zhao, Rongbao; Goldman, I. David.

In: Cancer Chemotherapy and Pharmacology, Vol. 72, No. 3, 09.2013, p. 597-606.

Research output: Contribution to journalArticle

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abstract = "Purpose: To characterize, directly and for the first time, the membrane transport and metabolism of pralatrexate, a new-generation dihydrofolate reductase inhibitor approved for the treatment for peripheral T-cell lymphoma. Experimental design: [3H]pralatrexate transport was studied in unique HeLa cell lines that express either the reduced folate carrier (RFC) or the proton-coupled folate transporter (PCFT). Metabolism to active polyglutamate derivatives was assessed by liquid chromatography. These properties were compared to those of methotrexate (MTX). Results: The pralatrexate influx K t, mediated by RFC, the major route of folate/antifolate transport at systemic pH, was 0.52 μΜ, 1/10th the MTX influx K i. The electrochemical potential of pralatrexate within HeLa cells far exceeded the extracellular level and was greater than for MTX. In contrast, MTX transport mediated by PCFT, the mechanism of folate/antifolate absorption in the small intestine, exceeded that for pralatrexate. After a 6 h exposure of HeLa cells to 0.5 μM pralatrexate, 80 {\%} of intracellular drug was its active polyglutamate forms, predominantly the tetraglutamate, and was suppressed when cells were loaded with natural folates. There was negligible formation of MTX polyglutamates. The difference in pralatrexate and MTX growth inhibition was far greater after transient exposures (375-fold) than continuous exposure (25-fold) to the drugs. Conclusions: Pralatrexate's enhanced activity relative to MTX is due to its much more rapid rate of transport and polyglutamation, the former less important when the carrier is saturated. The low affinity of pralatrexate for PCFT predicts a lower level of enterohepatic circulation and increased fecal excretion of the drug relative to MTX.",
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AU - Goldman, I. David

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