Profiling at recombinant homomeric and heteromeric rat P2X receptors identifies the suramin analogue NF449 as a highly potent P2X receptor antagonist

Rettinger, Jürgen; Braun, K.; Hochmann, Henrike; Kassack, Matthias U.; Ullmann, Heiko; Nickel, Peter; Schmalzing, Günther; Lambrecht, Günter

doi:10.1016/j.neuropharm.2004.11.003

Cited by 68 publications

(87 citation statements)

References 34 publications

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“…The nucleotides acted with halfmaximal concentrations in the submicromolar range compatible with an action on P2X 1 receptors. The interaction with the selective P2X 1 receptor antagonist NF449 used at the low concentrations of 100 nM and 1 μM [38,[41][42][43] clearly characterizes the involved subtype as P2X 1 receptor as discussed above. Rat coronary arteries express P2X 4 receptors [43,44] in addition to P2X 1 receptors.…”

Section: Discussionmentioning

confidence: 70%

“…First, α,β-methylene-ATP and β,γ-methylene-ATP induced the expression of the early gene NR4A1 in a manner sensitive to blockade of P2X 1 receptors by the selective antagonist NF449 [38,42,43]. And secondly, knockdown of the NR4A1 by treatment of cells with siRNA directed against the NR4A1 sequence attenuated the inhibitory effects of α,β-methylene-ATP and β,γ-methylene-ATP on cell proliferation.…”

Section: Discussionmentioning

confidence: 98%

“…NF449 is the most selective P2X 1 ligand available [41]. It was developed and thoroughly investigated in recombinant rat P2X receptors by Rettinger et al [42]. The following IC 50 values were obtained: 0.3 nM, 0.7 nM, 0.3 μM, 1.8 μM, and >300 μM for rP2X 1 , rP2X 1+5 , rP2X 2+3 , rP2X 3 , and rP2X 4 receptors, respectively.…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

P2X1 receptor-mediated inhibition of the proliferation of human coronary smooth muscle cells involving the transcription factor NR4A1

Hinze

Mayer

Harst

et al. 2013

Purinergic Signalling

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Adenine nucleotides acting at P2X 1 receptors are potent vasoconstrictors. Recently, we demonstrated that activation of adenosine A 2B receptors on human coronary smooth muscle cells inhibits cell proliferation by the induction of the nuclear receptor subfamily 4, group A, member 1 (NR4A1; alternative notation Nur77). In the present study, we searched for long-term effects mediated by P2X 1 receptors by analyzing receptor-mediated changes in cell proliferation and in the expression of NR4A1. Cultured human coronary smooth muscle cells were treated with selective receptor ligands. Effects on proliferation were determined by counting cells and measuring changes in impedance. The induction of transcription factors was assessed by qPCR. The P2X receptor agonist α,β-methylene-ATP and its analog β,γ-methylene-ATP inhibited cell proliferation by about 50 % after 5 days in culture with half-maximal concentrations of 0.3 and 0.08 μM, respectively. The effects were abolished or markedly attenuated by the P2X 1 receptor antagonist NF449 (carbonylbis-imino-benzenetriylbis-(carbonylimino)tetrakis-benzene-1,3-disulfonic acid; 100 nM and 1 μM). α,β-methylene-ATP and β,γ-methylene-ATP applied for 30 min to 4 h increased the expression of NR4A1; NF449 blocked or attenuated this effect. Small interfering RNA directed against NR4A1 diminished the antiproliferative effects of α,β-methylene-ATP and β,γ-methylene-ATP. α,β-methylene-ATP (0.1 to 30 μM) decreased migration of cultured human coronary smooth muscle cells in a chamber measuring changes in impedance; NF449 blocked the effect. In conclusion, our results demonstrate for the first time that adenine nucleotides acting at P2X 1 receptors inhibit the proliferation of human coronary smooth muscle cells via the induction of the early gene NR4A1.

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Section: Discussionmentioning

confidence: 70%

Section: Discussionmentioning

confidence: 98%

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P2X1 receptor-mediated inhibition of the proliferation of human coronary smooth muscle cells involving the transcription factor NR4A1

Hinze

Mayer

Harst

et al. 2013

Purinergic Signalling

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“…Suramin is a nonselective P2 purinergic antagonist 60 and NF023 is a suramin analog that specifically antagonizes the P2X1 receptor activity. 61 NF449 and PPNDS are also potent and selective P2X1 receptor inhibitors, 62,63 whereas reactive blue 2 antagonizes the P2Y receptor activity. 64 Other positive hits included cellular signaling and mitochondrial electron transport inhibitors, as well as antagonists of glutamate receptors ( Table 2).…”

Section: Positive Hits From the Pilot Screenmentioning

confidence: 99%

High-Throughput HIV–Cell Fusion Assay for Discovery of Virus Entry Inhibitors

Marin

Giroud

et al. 2015

ASSAY and Drug Development Technologies

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HIV-1 initiates infection by merging its envelope membrane with the target cell membrane, a process that is mediated by the viral Env glycoprotein following its sequential binding to CD4 and coreceptors, CXCR4 or CCR5. Although HIV-1 fusion has been a target for antiviral therapy, the virus has developed resistance to drugs blocking the CCR5 binding or Env refolding steps of this process. This highlights the need for novel inhibitors. Here, we adapted and optimized an enzymatic HIV-cell fusion assay, which reports the transfer of virus-encapsulated b-lactamase into the cytoplasm, to high-throughput screening (HTS) with a 384-well format. The assay was robustly performed in HTS format and was validated by the pilot screen of a small library of pharmacologically active compounds. Several hits identified by screening included a prominent cluster of purinergic receptor antagonists. Functional studies demonstrated that P2X1 receptor antagonists selectively inhibited HIV-1 fusion without affecting the fusion activity of an unrelated virus that enters cells through an endocytic route. The inhibition of HIV-cell fusion by P2X1 antagonists was not through downmodulation of the cell surface expression of CD4 or coreceptors, thus implicating P2X1 receptor in the HIV-1 fusion step. The ability of these antagonists to inhibit viruses regardless of their coreceptor (CXCR4 or CCR5) preference indicates that fusion is blocked at a late step downstream of coreceptor binding. A future large-scale screening campaign for HIV-1 fusion inhibitors, using the above functional readout, will likely reveal novel classes of inhibitors and suggest potential targets for antiviral therapy.

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“…Various directly-acting P2Y 12 receptor antagonists, including CT50547 and novel pyrazolidine-3,5-dione derivatives, are under development as antithrombotic agents (Scarborough et al 2001, Fretz et al 2005. The suramin derivative NF449 is selective for the P2X 1 receptor (Rettinger et al 2005). The selective P2X 2,3 /P2X 3 receptor antagonist A-317491 has antinociceptive properties and has been developed as a radioligand, but suffers from low bioavailability .…”

Section: Novel Non-nucleotide Antagonist Of P2 Receptorsmentioning

confidence: 99%

Agonists and Antagonists for P2 Receptors

Jacobson

Costanzi

Joshi

et al. 2006

Novartis Foundation Symposia

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Recent work has identified nucleotide agonists selective for P2Y 1 , P2Y 2 and P2Y 6 receptors and nucleotide antagonists selective for P2Y 1 , P2Y 12 and P2X 1 receptors. Selective non-nucleotide antagonists have been reported for P2Y 1 , P2Y 2 , P2Y 6 , P2Y 12 , P2Y 13 , P2X 2/3 /P2X 3 and P2X 7 receptors. For example, the dinucleotide INS 37217 (Up 4 dC) potently activates the P2Y 2 receptor, and the non-nucleotide antagonist A-317491 is selective for P2X 2/3 /P2X 3 receptors. Nucleotide analogues in which the ribose moiety is substituted by a variety of novel ring systems, including conformation-ally locked moieties, have been synthesized as ligands for P2Y receptors. The focus on conformational factors of the ribose-like moiety allows the inclusion of general modifications that lead to enhanced potency and selectivity. At P2Y 1,2,4,11 receptors, there is a preference for the North conformation as indicated with (N)-methanocarba analogues. The P2Y 1 antagonist MRS2500 inhibited ADP-induced human platelet aggregation with an IC 50 of 0.95 nM. MRS2365, an (N)-methanocarba analogue of 2-MeSADP, displayed potency (EC 50 ) of 0.4 nM at the P2Y 1 receptor, with >10 000-fold selectivity in comparison to P2Y 12 and P2Y 13 receptors. At P2Y 6 receptors there is a dramatic preference for the South conformation. Three-dimensional structures of P2Y receptors have been deduced from structure activity relationships (SAR), mutagenesis and modelling studies. Detailed three-dimensional structures of P2X receptors have not yet been proposed. Extracellular purine and pyrimidine nucleotides act as neurotransmitters/modulators . These ubiquitous signalling molecules modulate the function of diverse mammalian cell types and tissues under both normal and pathophysiological conditions. Receptors for extracellular nucleotides have been characterized through medicinal chemical, molecular biological, and pharmacological approaches. The eight subtypes of P2Y receptors, denoted P2Y 1 , P2Y 2 , P2Y 4 , P2Y 6 , P2Y 11 , P2Y 12 , P2Y 13 and P2Y 14 , are all seven transmembrane-spanning (7TM) receptors, which couple to G proteins. The seven P2X receptor subunits (P2X 1 -P2X 7 ) form multimeric ligand-gated ion channels. The distribution of P2Y receptors is broad, and the relevant therapeutic interests include antithrombotic therapy, modulation of the immune system and cardiovascular system, and treatment of cystic fibrosis and other pulmonary diseases (Yerxa et al 2002). Several of the

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Profiling at recombinant homomeric and heteromeric rat P2X receptors identifies the suramin analogue NF449 as a highly potent P2X receptor antagonist

Cited by 68 publications

References 34 publications

P2X1 receptor-mediated inhibition of the proliferation of human coronary smooth muscle cells involving the transcription factor NR4A1

P2X1 receptor-mediated inhibition of the proliferation of human coronary smooth muscle cells involving the transcription factor NR4A1

High-Throughput HIV–Cell Fusion Assay for Discovery of Virus Entry Inhibitors

Agonists and Antagonists for P2 Receptors

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