title: Synthesis and biological studies of novel nucleoside phosphoramidate prodrugs.
abstract: A novel approach to the intracellular delivery of nucleotides using phosphoramidate-based prodrugs is described. Specifically, we have developed phosphoramidate prodrugs of the anticancer nucleotide 5-fluoro-2'-deoxyuridine-5'monophosphate (FdUMP). These phosphoramidate prodrugs contain an ester group that undergoes intracellular activation liberating phosphoramidate anion, which undergoes spontaneous cyclization and P-N bond cleavage to yield the nucleoside monophosphate quantitatively. In vitro evaluation of 5-fluoro-2'-deoxyuridine phosphoramidate prodrugs 2a and 3b against L1210 mouse leukemia cells show potent inhibition of cell growth (IC(50) 0.5-3 nM). Cell-based thymidylate synthase inhibition studies show that, in contrast to FUdR, the nitrofuran compound 2a is of comparable potency in wild type vs thymidine kinase deficient LM cells. This result indicates that the activation of this novel prodrug occurs via the proposed mechanism of intracellular delivery. However, naphthoquinone 3b has an IC(50) value for thymidylate synthase inhibition that is comparable to FUdR in thymidine kinase deficient cells. Further studies revealed that 3b rapidly decomposes to the nucleotide in cell culture medium, suggesting that the naphthoquinone analogue is not sufficiently stable to function as a nucleotide prodrug.
authors.affiliation: Department of Medicinal Chemistry and Molecular Pharmacology and the Cancer Center, Purdue University, West Lafayette, Indiana 47907, USA.
authors.forename: S C
authors.initials: SC
authors.lastname: Tobias
authors.forename: R F
authors.initials: RF
authors.lastname: Borch
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D004354:Drug Screening Assays, Antitumor; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005467:Floxuridine / Q000494:pharmacology; D005468:Fluorodeoxyuridylate / Q000031:analogs & derivatives* / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D011355:Prodrugs / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D013937:Thymidine Kinase / Q000172:deficiency; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors; D014407:Tumor Cells, Cultured; D014542:Uridine Monophosphate / Q000031:analogs & derivatives / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology
chemical_list: C442812:5-fluoro-2'-deoxyuridyl 2-(1,4-naphthoquinoyl)methyl N-methyl-N-(4-chlorobutyl)phosphoramidate; C442811:5-fluoro-2'-deoxyuridyl 5-nitrofurfuryl N-methyl-N-(4-chlorobutyl)phosphoramidate; D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D011355:Prodrugs; D005467:Floxuridine; D005468:Fluorodeoxyuridylate; D014542:Uridine Monophosphate; D013940:Thymidylate Synthase; D013937:Thymidine Kinase