PPADS and suramin as antagonists at cloned P<sub>2Y</sub>‐ and P<sub>2U</sub>‐purinoceptors

Charlton, Steven J.; Brown, Colin B.; Weisman, Gary A.; Turner, John T.; Erb, Laurie; Boarder, Michael R.

doi:10.1111/j.1476-5381.1996.tb15457.x

Cited by 135 publications

(109 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is also similar to the pA 2 value of 5.46 determined at the endogenous P2Y 1 receptor in the rat mesenteric arterial bed [Lambrecht, 1996]. At the cloned and expressed turkey P2Y 1 receptor, a higher pA 2 value of 5.98 was estimated for PPADS; however, in these experiments the slope of the Schild plot regression line differed from unity [Charlton et al, 1996]. It should be noted that we did not prove the competitive character of the interaction of MRS 2210, NF023, and Reactive blue 2 with the human P2Y 1 receptor; but the present results do also not argue against a competitive antagonism.…”

Section: Discussionsupporting

confidence: 46%

Evidence for the recognition of non‐nucleotide antagonists within the transmembrane domains of the human P2Y₁ receptor

Guo

Kügelgen

Moro

et al. 2002

Drug Development Research

View full text Add to dashboard Cite

Site-directed mutagenesis was used to search for amino acid residues of the human P2Y 1 receptor involved in the binding of the P2 receptor antagonists pyridoxal-5′-phosphate-6-azophenyl-2,4-disulfonate (PPADS), its analogue 6-(2′-chloro-phenylazo)-pyridoxal-α 5 -phosphate (MRS 2210), the suramin analogue 8-8′-[carbonylbis(imino-3,1-phenylene)]bis(1,3,5-naphthalene-trisulfonate) (NF023), and Reactive blue 2. Receptors containing single amino acid replacements at positions in transmembrane helical domains (TMs) 3, 5, 6, and 7 critical for the activation of the receptor by nucleotide agonists were expressed in COS-7 (African green monkey kidney) cells. Inositol phosphate accumulation was induced by 2-methylthioadenosine 5′-diphosphate (2-MeSADP). In wild type human P2Y 1 receptors, PPADS (10 to 60 µM), MRS 2210 (10 µM), NF023 (100 µM), and Reactive blue 2 (10 µM) shifted the concentration-response curve of 2-MeSADP in a parallel manner to the right. For PPADS, a pA 2 value of 5.2 was estimated. The shifts caused by MRS 2210, NF023, and Reactive blue 2 corresponded to apparent pK B values of 5.6, 5.0, and 5.8, respectively. In K280A mutant receptors, the affinities of PPADS, MRS 2210, NF023, and Reactive blue 2 were about 6-to 60-fold lower than those observed at wild type receptors. The K280A mutation also caused an approximately 1,000-fold increase in the EC 50 value of the agonist 2-MeSADP, similar to previous observations. In contrast, no major change in antagonistic potency was observed at receptors with other mutations in TMs 3, 5, 6, and 7. Thus, the residue Lys 280 (6.55), which is located within the upper third of TM 6 of the human P2Y 1 receptor, is not only critical for the activation of the receptor but also plays an important role in the binding of pyridoxal derivatives and a number of other chemically unrelated P2 receptor antagonists. Lys 280 seems to belong to an overlapping region of the respective binding sites.

show abstract

Section: Discussionsupporting

confidence: 46%

Evidence for the recognition of non‐nucleotide antagonists within the transmembrane domains of the human P2Y₁ receptor

Guo

Kügelgen

Moro

et al. 2002

Drug Development Research

View full text Add to dashboard Cite

show abstract

“…In addition to the blockade of various P2X receptors, PPADS also acts as an effective antagonist at the P2Y 1 receptor, whereas it is completely ineffective at the P2Y 11 receptor (Communi et al, 1999). PPADS does not recognize P2Y 2 receptors up to a concentration of 30 mM and is also a relatively ineffective antagonist at the uridine nucleotide-sensitive P2Y 4 and P2Y 6 receptors (Charlton et al, 1996;Bogdanov et al, 1998;Dol-Gleizes et al, 1999). PPADS also fails to block P2Y 12 receptor-mediated effects (Nicholas, 2001).…”

Section: Discussionmentioning

confidence: 99%

Stimulation of P2Y1 Receptors Causes Anxiolytic-like Effects in the Rat Elevated Plus-maze: Implications for the Involvement of P2Y1 Receptor-Mediated Nitric Oxide Production

Kittner

Franke

Fischer

et al. 2002

Neuropsychopharmacol

View full text Add to dashboard Cite

(ADPbS), the P2X 1,3 receptor agonist a,b-methylene ATP (a,bmeATP), the unspecific P2 receptor antagonist pyridoxalphosphate-6-azopheny l-2 0 ,4 0 -disulfonic acid (PPADS), and the specific P2Y 1 receptor antagonist N 6 -methyl-2 0 -deoxyadenosine-3 0 ,5 0 -bisphosphate (MRS 2179) on the elevated plus-maze behavior of the rat were investigated. All tested compounds were given intracerebroventricularly (0.5 ml). ADPbS (50 and 500 fmol) produced an anxiolytic-like behavioral profile reflected by an increase of the open arm exploration. The anxiolytic-like effects were antagonized by pretreatment with PPADS (5 pmol) or MRS 2179 (5 pmol). Both compounds caused anxiogenic-like effects when given alone. Furthermore, the anxiolytic-like effects of ADPbS could be antagonized by pretreatment with the nitric oxide synthase (NOS) inhibitor N w -nitro-L-arginine methyl ester (L-NAME). In addition, the anxiogenic-like effects of PPADS were reversed by the pretreatment with L-arginine (500 pmol), which is the natural substrate for NOS, but not by D-arginine (500 pmol), which is not. Immunofluorescence staining revealed the presence of P2Y 1 receptors on neurons in different brain regions such as hypothalamus, amygdala, hippocampus and the periaqueductal gray. Furthermore, the colocalization of P2Y 1 receptors and neuronal NOS (nNOS) on some neurons in these regions could be demonstrated. The highest density of P2Y 1 -and nNOS-immunoreactivity was detected in the dorsomedial hypothalamic nucleus. Taken together, the present results suggest that P2Y 1 receptors are involved in the modulation of anxiety in the rat. The anxiolytic-like effects after stimulation of P2Y 1 receptors seem to be in close connection with the related nitric oxide production.

show abstract

“…58,59 Interestingly, our screen identified a cluster of purinergic receptor inhibitors, suramin, NF023, NF449, PPNDS, and reactive blue 2 ( Table 2). 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.…”

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

View full text Add to dashboard Cite

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.

show abstract

PPADS and suramin as antagonists at cloned P_2Y‐ and P_2U‐purinoceptors

Cited by 135 publications

References 36 publications

Evidence for the recognition of non‐nucleotide antagonists within the transmembrane domains of the human P2Y₁ receptor

Evidence for the recognition of non‐nucleotide antagonists within the transmembrane domains of the human P2Y₁ receptor

Stimulation of P2Y1 Receptors Causes Anxiolytic-like Effects in the Rat Elevated Plus-maze: Implications for the Involvement of P2Y1 Receptor-Mediated Nitric Oxide Production

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

Contact Info

Product

Resources

About

PPADS and suramin as antagonists at cloned P2Y‐ and P2U‐purinoceptors

Cited by 135 publications

References 36 publications

Evidence for the recognition of non‐nucleotide antagonists within the transmembrane domains of the human P2Y1 receptor

Evidence for the recognition of non‐nucleotide antagonists within the transmembrane domains of the human P2Y1 receptor

Stimulation of P2Y1 Receptors Causes Anxiolytic-like Effects in the Rat Elevated Plus-maze: Implications for the Involvement of P2Y1 Receptor-Mediated Nitric Oxide Production

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

Contact Info

Product

Resources

About

PPADS and suramin as antagonists at cloned P_2Y‐ and P_2U‐purinoceptors

Evidence for the recognition of non‐nucleotide antagonists within the transmembrane domains of the human P2Y₁ receptor

Evidence for the recognition of non‐nucleotide antagonists within the transmembrane domains of the human P2Y₁ receptor