title: 5'-Phosphoramidates and 5'-diphosphates of 2'-O-allyl-beta-D-arabinofuranosyluracil, -cytosine, and -adenine: inhibition of ribonucleotide reductase.
abstract: Continuing our studies on ribonucleotide reductase (RNR) mechanism-based inhibitors, we have now prepared the diphosphates (DP) of 2'-O-allyl-1-beta-D-arabinofuranosyl-uracil and -cytosine and 2'-O-allyl-9-beta-D-arabinofuranosyl-adenine and evaluated their inhibitory activity against recombinant murine RNR. 2'-O-Allyl-araUDP proved to be inhibitory to RNR at an IC(50) of 100 microM, whereas 2'-O-allyl-araCDP was only marginally active (IC(50) 1 mM) and 2'-O-allyl-araADP was completely inactive. The susceptibility of the parent nucleosides to phosphorylation by thymidine kinase and 2'-deoxycytidine kinase was also investigated, and all nucleosides proved to be poor substrates for the above-cited kinases. Moreover, prodrugs of 2'-O-allyl-araU and -araC monophosphates, namely 2'-O-allyl-5'-(phenylethoxy-L-alanyl phosphate)-araU and -araC, were prepared and tested against tumor cell proliferation but proved to be inactive. A molecular modeling study has been conducted in order to explain our results. The data confirm that for both the natural and analogue nucleoside diphosphates, the principal determinant interaction with the active site of RNR is with the diphosphate group, which forms strong hydrogen bonds with Glu623, Thr624, Ser625, and Thr209. Our findings indicate that the poor phosphorylation may represent an explanation for the lack of marked in vitro cytostatic activity of the test compounds.
authors.affiliation: Department of Pharmaceutical Sciences, Ferrara University, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Manfredini
authors.forename: P G
authors.initials: PG
authors.lastname: Baraldi
authors.forename: E
authors.initials: E
authors.lastname: Durini
authors.forename: S
authors.initials: S
authors.lastname: Vertuani
authors.forename: J
authors.initials: J
authors.lastname: Balzarini
authors.forename: E
authors.initials: E
authors.lastname: De Clercq
authors.forename: A
authors.initials: A
authors.lastname: Karlsson
authors.forename: V
authors.initials: V
authors.lastname: Buzzoni
authors.forename: L
authors.initials: L
authors.lastname: Thelander
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D001086:Arabinofuranosyluracil / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D003561:Cytarabine / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D003842:Deoxycytidine Kinase / Q000737:chemistry; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D010766:Phosphorylation; D011355:Prodrugs / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors; D012264:Ribonucleotide Reductases / Q000037:antagonists & inhibitors*; D013329:Structure-Activity Relationship; D013937:Thymidine Kinase / Q000737:chemistry; D014407:Tumor Cells, Cultured; D014740:Vidarabine / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology
chemical_list: D000970:Antineoplastic Agents; D011355:Prodrugs; D011994:Recombinant Proteins; D003561:Cytarabine; D001086:Arabinofuranosyluracil; D012264:Ribonucleotide Reductases; D013937:Thymidine Kinase; D003842:Deoxycytidine Kinase; D014740:Vidarabine