title: NF546 [4,4'-(carbonylbis(imino-3,1-phenylene-carbonylimino-3,1-(4-methyl-phenylene)-carbonylimino))-bis(1,3-xylene-alpha,alpha'-diphosphonic acid) tetrasodium salt] is a non-nucleotide P2Y11 agonist and stimulates release of interleukin-8 from human monocyte-derived dendritic cells.
abstract: The G protein-coupled P2Y(11) receptor is involved in immune system modulation. In-depth physiological evaluation is hampered, however, by a lack of selective and potent ligands. By screening a library of sulfonic and phosphonic acid derivatives at P2Y(11) receptors recombinantly expressed in human 1321N1 astrocytoma cells (calcium and cAMP assays), the selective non-nucleotide P2Y(11) agonist NF546 [4,4'-(carbonylbis(imino-3,1-phenylene-carbonylimino-3,1-(4-methyl-phenylene)carbonylimino))-bis(1,3-xylene-alpha,alpha'-diphosphonic acid) tetrasodium salt] was identified. NF546 had a pEC(50) of 6.27 and is relatively selective for P2Y(11) over P2Y(1), P2Y(2), P2Y(4), P2Y(6), P2Y(12), P2X(1), P2X(2), and P2X(2)-X(3). Adenosine-5'-O-(3-thio)triphosphate (ATPgammaS), a nonhydrolyzable analog of the physiological P2Y(11) agonist ATP, and NF546 use a common binding site as suggested by molecular modeling studies and their competitive behavior toward the nanomolar potency antagonist NF340 [4,4'-(carbonylbis(imino-3,1-(4-methyl-phenylene)carbonylimino))bis(naphthalene-2,6-disulfonic acid) tetrasodium salt] in Schild analysis. The pA(2) of NF340 was 8.02 against ATPgammaS and 8.04 against NF546 (calcium assays). NF546 was further tested for P2Y(11)-mediated effects in monocyte-derived dendritic cells. Similarly to ATPgammaS, NF546 led to thrombospondin-1 secretion and inhibition of lipopolysaccharide-stimulated interleukin-12 release, whereas NF340 inhibited these effects. Further, for the first time, it was shown that ATPgammaS or NF546 stimulation promotes interleukin 8 (IL-8) release from dendritic cells, which could be inhibited by NF340. In conclusion, we have described the first selective, non-nucleotide agonist NF546 for P2Y(11) receptors in both recombinant and physiological expression systems and could show a P2Y(11)-stimulated IL-8 release, further supporting the immunomodulatory role of P2Y(11) receptors.
authors.affiliation: Institute of Pharmaceutical and Medicinal Chemistry, Pharmaceutical Biochemistry, Heinrich-Heine-University of Duesseldorf, Duesseldorf, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Meis
authors.forename: Alexandra
authors.initials: A
authors.lastname: Hamacher
authors.forename: Darunee
authors.initials: D
authors.lastname: Hongwiset
authors.forename: Claudia
authors.initials: C
authors.lastname: Marzian
authors.forename: Michael
authors.initials: M
authors.lastname: Wiese
authors.forename: Niels
authors.initials: N
authors.lastname: Eckstein
authors.forename: Hans-Dieter
authors.initials: HD
authors.lastname: Royer
authors.forename: Didier
authors.initials: D
authors.lastname: Communi
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Boeynaems
authors.forename: Ralf
authors.initials: R
authors.lastname: Hausmann
authors.forename: Günther
authors.initials: G
authors.lastname: Schmalzing
authors.forename: Matthias U
authors.initials: MU
authors.lastname: Kassack
mesh_terms: D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000378:metabolism; D003713:Dendritic Cells / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D004164:Diphosphonates / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism*; D008024:Ligands; D009282:Naphthalenesulfonates / Q000494:pharmacology*; D011485:Protein Binding; D058910:Purinergic P2 Receptor Agonists; D018048:Receptors, Purinergic P2 / Q000378:metabolism; D058469:Receptors, Purinergic P2X; D011994:Recombinant Proteins; D014162:Transfection
chemical_list: C546599:4,4'-(carbonylbis(imino-3,1-phenylenecarbonylimino-3,1-(4-methylphenylene)carbonylimino))bis(1,3-xylene-alpha,alpha'-diphosphonic acid); D004164:Diphosphonates; D016209:Interleukin-8; D008024:Ligands; D009282:Naphthalenesulfonates; C109734:P2RY11 protein, human; D058910:Purinergic P2 Receptor Agonists; D018048:Receptors, Purinergic P2; D058469:Receptors, Purinergic P2X; D011994:Recombinant Proteins; D000242:Cyclic AMP; D002118:Calcium