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4-Alkyloxyimino-cytosine nucleotides: tethering approaches to molecular probes for the P2Y 6 receptor.
- Source :
-
MedChemComm [Medchemcomm] 2013 May 30; Vol. 4, pp. 1156-1165. - Publication Year :
- 2013
-
Abstract
- 4-Alkyloxyimino derivatives of pyrimidine nucleotides display high potency as agonists of certain G protein-coupled P2Y receptors (P2YRs). In an effort to functionalize a P2Y <subscript>6</subscript> R agonist for fluorescent labeling, we probed two positions ( N <superscript>4</superscript> and γ-phosphate of cytidine derivatives) with various functional groups, including alkynes for click chemistry. Functionalization of extended imino substituents at the 4 position of the pyrimidine nucleobase of CDP preserved P2Y <subscript>6</subscript> R potency generally better than γ-phosphoester formation in CTP derivatives. Fluorescent Alexa Fluor 488 conjugate 16 activated the human P2Y <subscript>6</subscript> R expressed in 1321N1 human astrocytoma cells with an EC <subscript>50</subscript> of 9 nM, and exhibited high selectivity for this receptor over other uridine nucleotide-activated P2Y receptors. Flow cytometry detected specific labeling with 16 to P2Y <subscript>6</subscript> R-expressing but not to wild-type 1321N1 cells. Additionally, confocal microscopy indicated both internalized 16 ( t <subscript>1/2</subscript> of 18 min) and surface-bound fluorescence. Known P2Y <subscript>6</subscript> R ligands inhibited labeling. Theoretical docking of 16 to a homology model of the P2Y <subscript>6</subscript> R predicted electrostatic interactions between the fluorophore and extracellular portion of TM3. Thus, we have identified the N <superscript>4</superscript> -benzyloxy group as a structurally permissive site for synthesis of functionalized congeners leading to high affinity molecular probes for studying the P2Y <subscript>6</subscript> R.
Details
- Language :
- English
- ISSN :
- 2040-2503
- Volume :
- 4
- Database :
- MEDLINE
- Journal :
- MedChemComm
- Publication Type :
- Academic Journal
- Accession number :
- 26161252
- Full Text :
- https://doi.org/10.1039/C3MD00132F