4-Alkyloxyimino derivatives of uridine-5'-triphosphate: distal modification of potent agonists as a strategy for molecular probes of P2Y2, P2Y4, and P2Y6 receptors.
Extended N(4)-(3-arylpropyl)oxy derivatives of uridine-5'-triphosphate were synthesized and potently stimulated phospholipase C stimulation in astrocytoma cells expressing G protein-coupled human (h) P2Y receptors (P2YRs) activated by UTP (P2Y2/4R) or UDP (P2Y6R). The potent P2Y4R-selective N(4)-(3-phenylpropyl)oxy agonist was phenyl ring-substituted or replaced with terminal heterocyclic or naphthyl rings with retention of P2YR potency. This broad tolerance for steric bulk in a distal region was not observed for dinucleoside tetraphosphate agonists with both nucleobases substituted. The potent N(4)-(3-(4-methoxyphenyl)-propyl)oxy analogue 19 (EC50: P2Y2R, 47 nM; P2Y4R, 23 nM) was functionalized for chain extension using click tethering of fluorophores as prosthetic groups. The BODIPY 630/650 conjugate 28 (MRS4162) exhibited EC50 values of 70, 66, and 23 nM at the hP2Y2/4/6Rs, respectively, and specifically labeled cells expressing the P2Y6R. Thus, an extended N(4)-(3-arylpropyl)oxy group accessed a structurally permissive region on three Gq-coupled P2YRs, and potency and selectivity were modulated by distal structural changes. This freedom of substitution was utilized to design of a pan-agonist fluorescent probe of a subset of uracil nucleotide-activated hP2YRs.
Molecular Recognition Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health , Bethesda, Maryland 20892 United States.
P Suresh
PS
Jayasekara
Matthew O
MO
Barrett
Christopher B
CB
Ball
Kyle A
KA
Brown
Eva
E
Hammes
Ramachandran
R
Balasubramanian
T Kendall
TK
Harden
Kenneth A
KA
Jacobson
D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D007097:Imines / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D015335:Molecular Probes; D058910:Purinergic P2 Receptor Agonists / Q000737:chemistry* / Q000494:pharmacology; D058473:Receptors, Purinergic P2Y2 / Q000737:chemistry / Q000145:classification / Q000187:drug effects*; D014544:Uridine Triphosphate / Q000737:chemistry*
D005456:Fluorescent Dyes; D007097:Imines; D015335:Molecular Probes; D058910:Purinergic P2 Receptor Agonists; D058473:Receptors, Purinergic P2Y2; D014544:Uridine Triphosphate