The effect of a novel transition state inhibitor of methylthioadenosine phosphorylase on pemetrexed activity.

Abstract

Pemetrexed is a new-generation antifolate inhibitor of thymidylate synthase (TS) and a weaker inhibitor of glycinamide ribonucleotide transformylase (GARFT) required for de novo purine synthesis. Methylthioadenosine phosphorylase (MTAP) salvages purines by releasing adenine from methylthioadenosine and is often deleted in mesothelioma. The current study addresses the effect of MTAP on pemetrexed activity using a highly potent transition state inhibitor of MTAP, MT-DADMe-Immucillin A (ImmA; K(i) = 86 pmol/L) in the MTAP(+) NCI-H28 and MTAP(-) NCI-H2052 mesothelioma cell lines. Based on selective nucleoside protection, TS was found to be the primary pemetrexed target in both cell lines with GARFT inhibition requiring 20- to 30-fold higher pemetrexed concentrations. ImmA had no effect on pemetrexed activity but, when thymidine was added, the pemetrexed IC(50) decreased by a factor of approximately 3 in MTAP(+) H28 cells with no effect in MTAP(-) H2052 cells. Conversely, the transfection of MTAP into H2052 cells increased the pemetrexed IC(50) by nearly 3-fold but only in the presence of thymidine; this was reversed by ImmA. An MTAP-specific short interfering RNA produced a 2-fold decrease in pemetrexed IC(50) in MTAP(+) HeLa cells in the presence of thymidine. These data indicate that suppression of constitutive MTAP has no effect on pemetrexed activity when the primary target is TS. There is a modest salutary effect when the pemetrexed target is GARFT alone.

• Authors: Shrikanta Chattopadhyay, Rongbao Zhao, Eugenia Tsai, Vern L Schramm, I David Goldman
• Journal: Molecular cancer therapeutics (Mol Cancer Ther) Vol. 5 Issue 10 Pg. 2549-55 (Oct 2006) ISSN: 1535-7163 [Print] United States
• PMID: 17041099 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Glutamates
• Pemetrexed
• Guanine
• Phosphoribosylglycinamide Formyltransferase
• Purine-Nucleoside Phosphorylase
• 5'-methylthioadenosine phosphorylase

Topics

• Cell Line, Tumor
• Glutamates (pharmacology)
• Guanine (analogs & derivatives, pharmacology)
• Humans
• Pemetrexed
• Phosphoribosylglycinamide Formyltransferase (antagonists & inhibitors)
• Purine-Nucleoside Phosphorylase (antagonists & inhibitors, biosynthesis, genetics)
• Reverse Transcriptase Polymerase Chain Reaction
• Transfection
• Tumor Stem Cell Assay