title: Development of ethynyl-2'-deoxyuridine chemical probes for cell proliferation.
abstract: A common method of evaluating cellular proliferation is to label DNA with chemical probes. 5-Ethynyl-2'-deoxyuridine (EdU) is a widely utilized chemical probe for labeling DNA, and upon incorporation, EdU treatment of cells is followed by a reaction with a small molecule fluorescent azide to allow detection. The limitations when using EdU include cytotoxicity and a reliance on nucleoside active transport mechanisms for entry into cells. Here we have developed six novel EdU pro-labels that consist of EdU modified with variable lipophilic acyl ester moieties. This pro-label:chemical probe relationship parallels the prodrug:drug relationship that is employed widely in medicinal chemistry. EdU and EdU pro-labels were evaluated for their labeling efficacy and cytotoxicity. Several EdU pro-label analogues incorporate into DNA at a similar level to EdU, suggesting that nucleoside transporters can be bypassed by the pro-labels. These EdU pro-labels also had reduced toxicity compared to EdU.
authors.affiliation: Discovery Biology, Eskitis Institute for Drug Discovery, Griffith University, Don Young Road, Nathan, Queensland 4111, Australia.
authors.forename: Carrie J
authors.initials: CJ
authors.lastname: Lovitt
authors.affiliation: Eskitis Institute for Drug Discovery, Griffith University, Don Young Road, Nathan, Queensland 4111, Australia.
authors.forename: David H
authors.initials: DH
authors.lastname: Hilko
authors.affiliation: Discovery Biology, Eskitis Institute for Drug Discovery, Griffith University, Don Young Road, Nathan, Queensland 4111, Australia.
authors.forename: Vicky M
authors.initials: VM
authors.lastname: Avery
authors.affiliation: Eskitis Institute for Drug Discovery, Griffith University, Don Young Road, Nathan, Queensland 4111, Australia. Electronic address: s.poulsen@griffith.edu.au.
authors.forename: Sally-Ann
authors.initials: SA
authors.lastname: Poulsen
mesh_terms: D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004247:DNA / Q000235:genetics; D003857:Deoxyuridine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology* / Q000633:toxicity; D004952:Esters / Q000138:chemical synthesis / Q000494:pharmacology / Q000633:toxicity; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000494:pharmacology* / Q000633:toxicity; D057809:HEK293 Cells; D006801:Humans; D015335:Molecular Probes / Q000138:chemical synthesis / Q000235:genetics / Q000494:pharmacology* / Q000633:toxicity; D013329:Structure-Activity Relationship
chemical_list: D004952:Esters; D005456:Fluorescent Dyes; D015335:Molecular Probes; D004247:DNA; D003857:Deoxyuridine
keywords: Cell proliferation; Chemical probe; DNA replication; Nucleoside; Pro-label; Prodrug; Thymidine