title: Influence of the nucleobase and anchimeric assistance of the carboxyl acid groups in the hydrolysis of amino acid nucleoside phosphoramidates.
abstract: Nucleoside phosphoramidates (NPs) are a class of nucleotide analogues that has been developed as potential antiviral/antitumor prodrugs. Recently, we have shown that some amino acid nucleoside phosphoramidates (aaNPs) can act as substrates for viral polymerases like HIV-1 RT. Herein, we report the synthesis and hydrolysis of a series of new aaNPs, containing either natural or modified nucleobases to define the basis for their differential reactivity. Aqueous stability, kinetics, and hydrolysis pathways were studied by NMR spectroscopy at different solution pD values (5-7) and temperatures. It was observed that the kinetics and mechanism (P-N and/or P-O bond cleavage) of the hydrolysis reaction largely depend on the nature of the nucleobase and amino acid moieties. Aspartyl NPs were found to be more reactive than Gly or β-Ala NPs. For aspartyl NPs, the order of reactivity of the nucleobase was 1-deazaadenine>7-deazaadenine>adenine>thymine≥3-deazaadenine. Notably, neutral aqueous solutions of Asp-1-deaza-dAMP degraded spontaneously even at 4 °C through exclusive P-O bond hydrolysis (a 50-fold reactivity difference for Asp-1-deaza-dAMP vs. Asp-3-deaza-dAMP at pD 5 and 70 °C). Conformational studies by NMR spectroscopy and molecular modeling suggest the involvement of the protonated N3 atom in adenine and 1- and 7-deazaadenine in the intramolecular catalysis of the hydrolysis reaction through the rare syn conformation.
authors.affiliation: Laboratory of Medicinal Chemistry, Rega Institute for Medical Research, Katholieke Universiteit Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium.
authors.forename: Munmun
authors.initials: M
authors.lastname: Maiti
authors.forename: Servaas
authors.initials: S
authors.lastname: Michielssens
authors.forename: Natalia
authors.initials: N
authors.lastname: Dyubankova
authors.forename: Mohitosh
authors.initials: M
authors.lastname: Maiti
authors.forename: Eveline
authors.initials: E
authors.lastname: Lescrinier
authors.forename: Arnout
authors.initials: A
authors.lastname: Ceulemans
authors.forename: Piet
authors.initials: P
authors.lastname: Herdewijn
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000737:chemistry; D000577:Amides / Q000737:chemistry*; D000596:Amino Acids; D000998:Antiviral Agents / Q000138:chemical synthesis* / Q000737:chemistry; D018360:Crystallography, X-Ray; D015497:HIV-1 / Q000187:drug effects; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008962:Models, Theoretical; D008968:Molecular Conformation; D009705:Nucleosides / Q000737:chemistry*; D010756:Phosphoric Acids / Q000737:chemistry*; D011355:Prodrugs / Q000138:chemical synthesis* / Q000737:chemistry; D013329:Structure-Activity Relationship
chemical_list: D000577:Amides; D000596:Amino Acids; D000998:Antiviral Agents; D009705:Nucleosides; D010756:Phosphoric Acids; D011355:Prodrugs; C043837:7-deazaadenine; C055133:3-deazaadenine; C011067:phosphoramidic acid; D000225:Adenine